730R82101
MANUAL OF CHEMICAL METHODS
FOR PESTICIDES AND DEVICES
U.S. Environmental
Protection Agency
OFFICE OF PESTICIDE PROGRAMS
CHEMICAL AND BIOLOGICAL INVESTIGATIONS BRANCH
BELTSVILLE, MD
Published and Distributed by
The Association of Official Analytical Chemists
First Edition: July 1976
First Update: July 1977
Second Update: May 1979
Third Update: August 1982
U.S. Environmental Protection Agency
Region V, Librr.ry
230 South Cccuborn Slreet
Chicago, Illinois 60604
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Published and Distributed by
The Association of Official Analytical Chemists
1111 N. 19th Street, Suite 210, Arlington, VA 22209 USA
Phone 703/522-3032
U,S. Environmental Protection Agency
ISBN 0-935584-23-4
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Preface
The EPA Manual of Chemical Methods for Pesticides and Devices is a
compendium of analytical procedures for technical and commercial pesticide
formulations. It is an example of federal and state cooperation which
developed through the professional concern of scientists and scientific groups
who recognized that uniform, reliable, standardized analytical chemical
methods will help regulatory officials better serve the public as well as the
c regulated industries.
The initial plan to publish a manual originated with the EPA's
Beltsville, Maryland laboratory scientists. The idea was proposed to the
Methods Clearing House Committee of the Association of American Pest Control
Official (AAPCO) for their consideration. In August 1974, at their 28th
Annual Meeting, AAPCO passed a resolution requesting EPA's Technical Services
Division (TSD) to prepare and maintain a manual of methods for pesticide
formulation analysis. In October of the same year, EPA, TSD's Methods
Development Coordination chemists (Beltsville, Maryland) and AAPCO's Methods
Clearing House Committee (AAPPCO's official body designed to work with EPA)
held a meeting to decide on the general format and contents of the proposed
manual. In July of 1976, the first edition of the manual was published.
Continuing with the aim of providing analytical methods which can be used
to support enforcement actions, the editors decided to reprint the original
~ manual (July 1976) ; the updates of 1977 and 1979; and 55 additional methods.
This 2nd edition EPA Manual of Chemical Methods for Pesticides and Devices
contains 317 analytical methods for 162 chemicals which may be found in
-r commercial pesticide formulations.
• Although the procedures in this manual have not achieved official AOAC
^ status through collaborative testing, most have been partially validated in
the EPA and state laboratories. In many instances, the procedures are
believed to be the best and, in some cases, the only methods available for a
particular formulation.
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It is hoped that these methods will eventually achieve official AOAC status by
collaborative study under the direction of AOAC associate referees. If a
method achieves official status, it will be published in the AOAC Methods of
Analysis and deleted from this manual. All methods will be reviewed by a
committee of AOAC referees and AAPCO Clearing House Committee members. The
corrmittee will recommend to the editors appropriate actions with respect to
corrections, modification, and deletions.
The AOAC recognizes the method gap which exists because of the limited
number of applicable official analyses available for analyzing the many
pesticide products sold to the public. Therefore, it is pleased to join the
EPA and AAPCO through the publication and sale of this manual in making
available to industry, states, other nations, academic and scientific
institutions, libraries, and the general public these analytical methods
currently being used by EPA and state laboratories in pesticide regulatory
programs. Where an official AOAC method does exist it is, of course, the
method of choice.
The editors wish to thank those people who have contributed in making
this manual possible. The editors would also appreciate receiving methods and
suggestions for making the manual a viable and continuing source of
methodology.
Editors: Warren R. Bontoyan
Jack B. Looker
Chemical and Biological
Investigations Branch
Environmental Protection Agency
BuiIding 402, ARC-East
Beltsville, MD 20705
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Preface
This EPA Manual of Chemical Methods for Pesticides and Devices
is a compendium of over 200 analytical procedures for commercial
pesticide formulations. It also contains 350 infrared curves and a
bibliography of books, manuals, and periodicals relating to pesticides.
The initial plan to publish a manual originated within EPA and
the idea was proposed to the Methods Clearing House Committee of the
Association of American Pesticide Control Officials (AAPCO) for their
consideration. In August 1974, at their 28th Annual Meeting, AAPCO
passed a resolution requesting EPA's Technical Services Division (TSD)
to prepare and maintain a manual of methods for pesticide formulation
analysis. In October of the same year, EPA, TSD's Methods Develop-
ment Coordination chemists, and AAPCO's Methods Clearing House Committee
(AAPCO's official body designated to work with EPA) held a meeting to
decide on the general format and content of the proposed manual. Also
at that time two committees were formed: (1) an Editorial Committee
(consisting of k EPA pesticide formulation chemists and 2 state chemists
recognized by AAPCO as having experience and expertise in formulation
analysis) whose task would be to standardize the method format and edit
all related material to be included in the manual; (2) a Method Review
Committee (comprised of a majority of experienced state chemists and a
minority of experienced EPA formulation chemists) having the responsibility
for accepting or rejecting analytical methods submitted for inclusion in
the manual. The present members of these committees are listed at the end
of this preface.
Many of the methods in the manual have been reviewed and accepted by
the committee. Some were not reviewed but were accepted because of their
wide use (e.g. Virginia Department of Agriculture Methods and Mississippi
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State Chemical Laboratory Methods). This procedure was agreed to by
AAPCO and EPA in October 1975. Also, it was agreed that certain methods
be designated as "Tentative." This designation was chosen for new
techniques or experimental methods and for those methods not widely used.
However, after one year in the manual, these tentative methods will be
submitted to the Method Review Committee for a final decision of full
acceptance (removal of "Tentative" designation) or rejection.
Analytical methods are currently being developed at a rapid pace,
and procedures and data are being generated at a rate much faster than
they can be validated. Although the procedures in this manual have not
achieved official AOAC status through collaborative testing, most of
them have been partially validated in the EPA and state laboratories.
In many instances, the procedures are believed to be the best—in some
cases the only—methods available for a particular formulation. It is
hoped that the manual methods will eventually achieve official AOAC
status by collaborative study under the direction of AOAC Associate
Referees. When a method does receive official status, it will be deleted
from the manual. The loose-leaf format was chosen to facilitate both this
deletion and the addition of new or improved methods and data. Semiannual
updates will be issued to keep the manual current and as free from error
as possible. The "devices" mentioned in the title of this volume, although
not included in the original issue, will appear in future updates.
This manual is an example of Federal and state cooperation that
developed through the professional concern of individuals and groups for
standardizing chemical analyses used by Federal and state pesticide
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regulatory laboratories, and the recognition that uniform, reliable
chemical methods will help regulatory officials better serve the
public as well as the regulated industries.
The AOAC has recognized the gap that exists because of the
limited number of applicable official methods; therefore, it is pleased
to join with EPA and AAPCO through the publication and sale of this
manual to make available to industry, academic and scientific institu-
tions, libraries, and the public these analytical procedures currently
being used by EPA and state laboratories in enforcing the law. Where
an official AOAC procedure does exist, it is, of course, the method of
choice.
The Editorial Chairman would appreciate receiving corrections,
suggestions, and new and improved methods or data for inclusion in
this manual. He will forward the methods (after conversion to standard
format) and pertinent comments to the chairman of the AAPCO-EPA Review
Committee for appropriate action.
The compilers of this manual take this opportunity to thank those
who have helped collect the information presented in this volume and to
request their help and the help of others in maintaining the manual as
a viable and current source of methodology.
Warren Bontoyan
Chairman of the Editorial Committee
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AAPCa-EPA, Editorial Committee
Warren R. Bontoyan, Chairman
EPA, TSD
Bldg. 306, Rra 101, Beltsville, Md. 20705
•
Coleman Hall, Assistant Chairman - EPA, TSD
Jack Looker, Assistant Chairman - EPA, TSD
Bernard Gildea - EPA, Region II
Leo Cox - Consolidated Labs, Virginia
J. Ron Conley - Ga. Dept. Agric., Georgia
AAPCO-EPA Review Committee
Paul Irwin, Chairman
Division of Consolidated Laboratory Services
1 North 14th Street
Richmond, Virginia 23219
Joseph Audino - California
Gregory Beierl - EPA
Warren Bontoyan - EPA
E. L. Campbell - Alberta, Canada
W. Y. Cobb - North Carolina
H. Kent Francis - Utah
Stelios Gerazounis - EPA
Alan Hanks - Texas
Virgil Hiatt - Oregon
Dean Hill - EPA
P. Stan Jones - Illinois
Paul Jung - Maryland
Daniel McDaniels - EPA
J. P. Minyard - Mississippi
E. R. Winterle - Florida
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Table of Contents
Preface
Methods of Analysis:
Pesticide Name Cross Reference Index to the Methods
Analytical Methods - Introduction
Methods
Thin Layer Chromatography
Bibliography
Infrared Spectra of Pesticides:
Introduction and Tabulation of Data
Index
Spectra
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ANALYTICAL METHODS
Pesticide Name Cross Reference Index to the Methods
Aatrex
ACC 3422
Accelerate
3-(alpha~acetonylbenzyl)-4-
hydroxycoumarin
3-(alpha-acetonylfurfuryl)-4-
hydroxycoumarin
Acricid
Afalon
Agrimycin
Agri-Strep
Agritox
Agrotect
Agroxone
Alachlor EPA-1 (tentative)
Alachlor EPA-2 (tentative)
Alkron
Aileron
Allisan
Ambox
Amcide
Amerol
5-amino-4-chloro-2-phenyl-
3(2H)-pyridazinone
Aminopyridine EPA-1 (tentative)
4-aminopyr id ine
aminotriazole
Atrazine EPA-1 & 2
Parathion EPA-1 & 2
Endothall EPA-1 & 2
Warfarin EPA-1, 2, & 3
Coumafuryl EPA-1 & 2
Binapacryl EPA-1
Linuron EPA-1 & 2
Streptomycin EPA-1
Streptomycin EPA-1
MCPA
2,4-D
MCPA
GLC-TCD-IS
GLC-FID-IS
Parathion EPA-1 & 2
Parathion EPA-1 & 2
Dicloran EPA-1
Binapacryl EPA-1
AMS EPA-1
Amitrole EPA-1
Pyrazon EPA-1
UV
Aminopyridine EPA-1
Amitrole EPA-1
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3-amino-s-triazole
3-amino-l,2,4-triazole
4-amino-3,5,6-trichloropicolinic acid
Amitrol EPA-1
Amizol
Ammate
ammonium methanearsonate
ammonium sulfamate
Amoxone
AMS EPA-1
p-tert-amylphenol
anilazine
Anilazine EPA-1 (tentative)
Anilazine EPA-2 (tentative)
anofex
Ansar
Antimilace
4-AP
Aphamite
Aqua-Keen
Aquathol
Aqua-Vex
Arasan
Arathane
Arsenic Compounds EPA-1
Arsenic Compounds EPA-2
Amitrole EPA-1
Amitrole EPA-1
Picloram EPA-1
Visible (colorimetric) spectroscopy
Amitrole EPA-1
AMS EPA-1
Arsenic Compounds EPA-3 & 4
AMS EPA-1
2,4-D
sodium nitrate titration
Phenols & Chlorophenols
EPA-1, 6, & 8
Chloro-Triazine Herbicides EPA-1
IR
GLC-TCD-IS
DDT EPA-1
Arsenic Compounds EPA-3 & 4
Metaldehyde EPA-1, 2, 3, & 4
Aminopyridine EPA-1
Parathion EPA-1 & 2
2,4-D
Endothall EPA-1 & 2
silvex
Thiram EPA-1 & 2
Dinocap EPA-1 & 2
iodometric titration
digestion, reduction, titration
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Arsenic Compounds EPA-3 (tentative)
Arsenic Compounds EPA-4
arsenic trioxide
Asuntol
ATA
Atlacide
Atranex
Atratole
atrazine
Atrazine EPA-1
Atrazine EPA-2 (tentative)
Avitrol
Azinphos-methyl EPA-1
Azak
Azodrin
Bay 21/199
B-622
Bay 17147
Bay 37344
Bay 70142
Bayer 19639
Baymix
Balan
Halfin
Banafin
Bansanite
digestion, reduction, titration
sulfuric acid digestion-iodine
titration
Arsenic Compounds EPA-1 & 2
Coumaphos EPA-1, 2, & 3
Amitrole EPA-1
Sodium Chlorate EPA-1
Atrazine EPA-1 & 2
Sodium Chlorate EPA-1
Chloro-Triazine Herbicides EPA-1
IR
GLC-FID-IS
Aminopyridine EPA-1
IR
Terbutol EPA-1 & 2
Monocrotophos EPA-1 & 2
Coumaphos EPA-1, 2, & 3
Anilazine EPA 1 & 2
Azinphos-methyl EPA-1
Methiocarb EPA-1
Carbofuran EPA-1
Disulfoton EPA-1 & 2
Coumaphos EPA-1, 2, & 3
Benefin EPA-1 & 2
Benefin EPA-1 & 2
Benefin EPA-1 & 2
Dinoseb EPA-1 & 2
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Barbacco (Spanish-speaking
Sc. Am. Countries)
Baron
Basfapon
Basudin
BBC 12
Benalin
Benefin EPA-1
Benefin EPA-2 (tentative)
benfluralin
Benlate
Benomyl EPA-1
Benomyl EPA-2 (tentative)
Bensulide EPA-1
Beosit
Benzahex
benzenehexachloride
Benzex
benzofuraline
o-benzyl-p-chlorophenol
2-benzyl-4-chlorophenol
(5-benzyl-3-furyl)methyl-2,2-
dimethyl-3-(2-methylpropenyl)
cyclopropanecarboxylate
Benzytol
Betasan
BHC, gamma isomer EPA-1
Binapacryl EPA-1 (tentative)
Rotenone
erbon
Dalapon EPA-1
Diazinon EPA-1, 2, 3, & 4
Dibromochloropropane EPA-1 & 2
Benefin EPA-1 & 2
IR
GLC-FID-IS
Benefin EPA-1 & 2
Benomyl EPA-1 & 2
IR
UV
IR
Endosulfan EPA-1, 2, 3, & 4
BHC, gamma isomer EPA-1
BHC, gamma isomer EPA-1
BHC, gamma isomer EPA-1
Resmethrin EPA-1, 2, 3, 4, & 5
Phenols & Chlorophenols
EPA-1, 3, 6, 7, & 8
o-benzyl-p-chlorophenol
Resmethrin EPA-1, 2, 3, 4, & 5
4-chloro-3,5-xylenol
Bensulide EPA-1
IR
IR
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Binnell
Benefin EPA-1 & 2
bls[2-(2,A-dlchlorophenoxy)
ethyl]phosphite
Bis(dimethylthiocarbamoyl)
disulphide
2,4-bis(isopropylamino)-6-
methoxy-s-triazine
bis(tributyltin) compounds
Bladan
Bladex
Blulan
Bonalan
boraclc acid
borax
Bordermaster
Borea
boric acid
Borocil
Borolin
Boron Compounds EPA-1
Botran
Bravo
Brimstone
Bromacil EPA-1 (tentative)
Bromex
Brominated Salicylanilides EPA-1
5-bromo-3-sec-butyl-6-roethyluracil
3-(4-bromo-3-chlorophenyl)-1-
methoxy-1-methylurea
2,A-DEP
Thiram EPA-1 & 2
Prometone EPA-1 & 2
Organotin Compounds EPA-1
Parathion EPA-1 & 2
Cyanazine EPA-1
Benefin EPA-1 & 2
Benefin EPA-1 & 2
Boron Compounds EPA-1
Boron Compounds EPA-1
MCPA
Bromacil EPA-1
Boron Compounds EPA-1
Bromacil EPA-1
Picloram EPA-1
ignition & titration
Dicloran EPA-1
Chlorothalonil EPA-1
Sulfur EPA-1, 2, & 3
GLC-FID-IS
Chlorbromuron EPA-1
UV
Bromacil EPA-1
Chlorbromuron EPA-1
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3-(p-bromophenyl)-l-inethoxy-l-
methylurea
Brush-Rhop
Butacide
Butoxone
a-[2-(2-n-butoxyethoxy)-ethoxy]-
4,5-methylenedioxy-2-propyltoluene
Butylate EPA-1 (tentative)
Butylate EPA-2 (tentative)
Butylate EPA-3 (tentative)
Butylate EPA-4
Butylate EPA-5 (tentative)
tert-butyl carbamic acid, ester with
3- (m-hydroxyphenyl) -1 , 1-d imethylurea
(butyl carbityl) (6-propylpiperonyl)
ether 80% and related compounds 20%
4-tert-butyl-2-chlorophenyl methyl
methylphosphoramidate
1-n-buty 1-3- ( 3 , 4-d ichlorophenyl ) -1-
methylurea
2-sec-butyl-4 , 6-dinitrophenol
2-sec-butyl-4 ,6-dinitrophenyl-3-
methyl-2-butenoate
2 ,6-dinitro-p-toluidine
Butylphen
p-tert-butylphenol
2-(p-tert-butylphenoxy)cyclohexyl
2-propynyl sulfite
2,6-di-tert-butyl-p-tolyl methyl-
carbamate
Metobrorauron EPA-1, 2, & 3
2,4,5-T
Piperonyl Butoxide EPA-1 & 2
2,4-DB
Piperonyl Butoxide EPA-1 & 2
GLC-TCD
HPLC
GLC-FID
GLC-FID-IS
GLC-TCD-IS
Karbutilate EPA-1
Piperonyl Butoxide EPA-1 & 2
Crufomate EPA-1 & 2
Neburon EPA-1
Dinoseb EPA-1 & 2
Binapacryl EPA-1
Benefin EPA-1 & 2
p-tert-butylphenol
Phenols & Chlorophenols
EPA-1, 6, & 8
Propargite EPA-1 & 2
Terbutol EPA-1 & 2
Butyrac
2,4-DB
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C-1983
C-2059
C-3126
cacodylic acid
cadmium carbonate
cadmium chloride
Cadmium Compounds EPA-1
cadmium oxide
cadmium sebacate
cadmium succinate
cadmium sulfate
calcium arsenate
calcium arsenite
Can-Trol
Captafol EPA-1 (tentative)
Captan EPA-1
Captan EPA-2
Carbaryl EPA-1
Carbaryl EPA-2 (tentative)
Carbofos
Carbofuran EPA-1
Carboxin EPA-1 (tentative)
Carfene
Carpidor
Casoron
Chemox
Chipco Turf Herbicide D
Chloroxuron EPA-1 & 2
Fluometuron EPA-1
Metobromuron EPA-1, 2, & 3
Arsenic Compounds EPA-3 & A
Cadmium Compounds EPA-1
Cadmium Compounds EPA-1
AA
Cadmium Compounds EPA-1
Cadmium Compounds EPA-1
Cadmium Compounds EPA-1
Cadmium Compounds EPA-1
Arsenic Compounds EPA-1 & 2
Arsenic Compounds EPA-1 & 2
MCPB
IR
hydrolyzable chlorine
D*
UV
HPLC
Malathion EPA-1 & 2
IR
IR
Azinphos-raethyl EPA-1
Benefin EPA-1 & 2
Dichlobenil EPA-1
Dinoseb EPA-1 & 2
2,4-D
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Chipco Turf Herbicide MCPP
Chiptox
2-chloro-4,6-bis(ethylamino)-s-
triazine
chlorobromuron (France)
Chlorbromuron EPA-1 (tentative)
4-chloro-2-cyclopentylphenol
2-chloro-2',6'-diethyl-N-
(methoxymethyl)acetanilide
4-chloro-3,5-dimethylphenol
2-chloro-4-ethylamino-6-isopropyl-
amino-1,3,5-triazine
2-(4-chloro-6~ethylamino-s-triazin-
2-ylamino)-2-methylpropionitrile
Chlorfenidim
Chlorfenizon
Chlorfension
2-chloro-5-hydroxy-l,3-dimethyl-
benzene
Chlorophene
Chlorophenothane
Chlorophenoxy Herbicides E_PA-1
Chlorophenoxy Herbicides EPA-2
Chlorophenoxy Herbicides EPA-3
(jentative)
Chlorophenoxy Herbicides EPA-4
(tentative)
Chlorophenoxy Herbicides EPA-5
(tentative)
3-[p-(p-chlorophenoxy)phenyl]-!,!-
dimethylurea
mecoprop
MCPA
Simazine EPA-1
Chlorbromuron EPA-1
GLC-FID
Phenols & Chlorophenols
EPA-1, 3, & 8
Alachlor EPA-1 & 2
4-chloro-3,5-xylenol
Atrazine EPA-1 & 2
Cyanazine EPA-1
Monuron EPA-1, 2, & 3
Ovex EPA-1
Ovex EPA-1
4-chloro-3,5-xylenol
o-benzyl-p-chlorophenol
DDT EPA-1
Definition, Structure, and
Technical Data
UV
HPLC
GLC-FID-IS
GLC-FID-IS (on column derivati-
zation)
Chloroxuron EPA-1 & 2
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p-chlorophenyl-p-chlorobenzene-
sulfonate
3-(p-chlorophenyl)-l,l-dimethylurea
4-chloro-2-phenylphenol
6-chloro-2-phenylphenol
Chlorothalontl EPA-1
Chlorothiepin
Chlpro^Triazine_Herbicides EPA-1
Chloroxifenidim
Chloroxone
Chloroxuron EPA-1 (tentative)
Chloroxuron EPA-2 (tentative)
p-chloro-m-xylenol
4-chloro-3,5-xylenol
CIBA-2059
cinerins
citral
CMPP
Comite
copper acetoarsenate
Co-Ral
Cornox M
Cornox RK
Corothion
Corotran
Cotnion-Methyl
Ovex EPA-1
Monuron EPA-1, 2, & 3
Phenols & Chlorophenols
EPA-1, 3, & 8
Phenols & Chlorophenols
EPA-1, 3, & 8
IR
Endosulfan EPA-1, 2, 3, & 4
chlorine potenttometrie titration
Chloroxuron EPA-1 & 2
2,4-D
IR
GLC-TCP-IS
4-chloro-3,5-xylenol
Phenols & Chlorophenols
EPA-1, 3, & 7
Fluometuron EPA-1
Pyrethrln EPA-1
Oil of Lemongrass EPA-1
mecoprop
Propargite EPA-1 & 2
Arsenic Compounds EPA-1 & 2
Coumaphos EPA-1, 2, & 3
MCPA
dichlorprop
Parathion EPA-1 & 2
Ovex EPA-1
Azinphos-methyl EPA-1
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10
Cotoran
coumafene (France)
Coumafuryl EPA-1
Coumafuryl EPA-2
Coumaphos EPA-1 (tentative)
Coumaphos EPA-2 (tentative)
Coumaphos EPA-3
CP 50144
CPCBS
Crop Rider
Crufomate EPA-1 (tentative)
Crufomate EPA-2 (tentative)
Crysan
Cube (Peru)
Curaterr
cyanazine
Cyanazine EPA-1
Cycloate EPA-1 (tentative)
Cycloate EPA-2 (tentative)
Cycloate EPA-3
Cyclodan
Cythion
Cytrol
2,4-D
D 735
Daconil 2787
Fluometuron EPA-1
Warfarin EPA-1, 2, & 3
UV (in baits)
IR (in concentrates)
IR
HPLC
GLC-FID-IS
Alachlor EPA 1 & 2
Ovex EPA-1
2,4-D
IR
GLC-TCD-IS
Resmethrin EPA-1, 2, 3, 4, & 5
Rotenone EPA-1
Carbofuran EPA-1
Chloro-Triazine Herbicides EPA-1
IR
GLC-TCD
GLC-FID
GLC-FID-IS
Endosulfan EPA-1, 2, 3, & 4
Malathion EPA-1 & 2
Amitrole EPA-1
Chlorophenoxy Herbicides
EPA-1, 2, 3, 4, & 5
Carboxin EPA-1
Chlorothalonil EPA-1
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11
Dalapon EPA-1
Dalf
Dazzel
2,4-DB
DBCP
2,6-DBN
DCMO
DCNA
DDT EPA-1
Decamine
De-Cut
Dedelo
Ded-Weed
Ded-Weed Brush Killer
Deet EPA-1 (tentative)
Deet EPA-2 (tentative)
Deet EPA-3 (tentative)
De-Fol-Ate
Delphene
2,4-DEP
Derris
Des-i-cate
De-Sprout
Detamide
Diazajet
IR
Methyl Parathion
EPA-1, 2, 3, 4, & 5
Diazinon EPA-1, 2, 3, & 4
Chlorophenoxy Herbicides
EPA-1, 2, & 3
Dibromochloropropane EPA-1 & 2
Dichlobenil EPA-1
Carboxin EPA-1
Dicloran EPA-1
IR
2,4-D & 2,4,5-T
MH EPA-1
DDT DPA-1
see 2,4-D, silvex, or dalapon
2,4,5-T
IR
GLC-TCD-IS
GLC-FID-IS
Sodium Chlorate EPA-1
Deet EPA-1, 2, & 3
Chlorophenoxy Herbicides
EPA-1, 2, & 3
Rotenone EPA-1
Endothall EPA-1 & 2
MH EPA-1
Deet EPA-1, 2, & 3
Diazinon EPA-1, 2, 3, & 4
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12
Diazide
Diazol
Diazinon EPA-1
Diazinon EPA-2 (tentatlye)
Diazinon EPA-3
Diazinon EPA-4
Dibromochloropropane EPA-1
Dibromochloropropane EPA-2 (tentative)
1,2-dibrotno-3-chloropropane
4',5-dibromosalicylanilides
Dibutyl Succinate EPA-1
Dichlobenil EPA-1
Dichlone EPA-1
dichlorfenidim
p-Dichlorobenzene EPA-1 (tentative)
p-Dichlprgbenzene EPA-2 (tentative)
1,4-dichlorobenzene
2,6-dichlorobenzonitrile
2,4-dichloro-6-(o-chloroanilino)-s-
triazine
4,6-dichloro-N-(2-chlorophenyl)-l,3,5-
triazin-2-amine
dichlorodiphenyltrichloroethane
2,3-dichloro-l,4-naphthoquinone
2,6-dichloro-4-nitroaniline
2,4-dichlorophenoxyacetic acid
4-(2,4-dichlorophenoxy)butyric acid
2-(2,4-dichlorophenoxy)propionic acid
Diazinon EPA-1, 2, 3, & 4
Diazinon EPA-1, 2, 3, & 4
GLC-TCD
HPLC
IE
GLC-FID-IS
IR
GLC-TCD
Dibroraochloropropane EPA-1 & 2
Brominated Salicylanilides EPA-1
saponification & titration
IR
IR
Diuron EPA-1, 2, 3, & 4
_m
GLC-TCD-IS
p-Dichlorobenzene EPA-1 & 2
Dichlobenil EPA-1
Anilazine EPA-1 & 2
Anilazine EPA 1 & 2
DDT EPA-1
Dichlone EPA-1
Dicloran EPA-1
2,4-D
2,4-DB
dichlorprop
-------
13
3-(3,4-dichlorophenyl)-l,l-
dimethylurea
3-(3,4-dichlorophenyl)-l-raethoxy-
1-methylurea
2,2-dichloropropionlc acid
dichlorprop
Dicloran EPA-1
Dicophane
Didimac
diethion
0,0-diethyl-0-(3-chloro-4-methyl-
2-OXO-2H-1-benzopyran-7-yl)
phosphorothioate
0,0-diethyl S [2-(ethylthio)ethyl]
phosphorodithioate
0,0-dlethyl S-(ethylthiomethyl)
phosphorodithioate
0,0-diethyl 0-(2-isopropyl-6-methyl-
4-pyrimidinyl)phosphorothioate
0,0-diethyl-O-p-nitrophenyl
phosphorothioate *
N ,N-diethyl-tn-toluamide
difenson
Difolatan
2,3-dihydro-2,2-dimethyl-7-
benzofuranyl methyl carbamate
5,6-dihydro-2-methyl-l,4-
oxathiin-3-carboxanilide
1,2 -d ihydro-pyr idaz ined ione
diisopropyl S-(2-phenylsul£onyl-
aminoethyl)phosphorothiolothionate
Diuron EPA-1, 2, 3, & 4
Linuron EPA-1 & 2
Dalapon EPA-1
Chlorophenoxy Herbicides
EPA-1, 2, & 3
IR
DDT EPA-1
DDT EPA-1
Ethion EPA-1 & 2
Coumaphos EPA-1, 2, & 3
Disulfoton EPA-1 & 2
Phorate EPA-1
Diazinon EPA-1, 2, 3, & 4
Parathion EPA-1 & 2
Deet EPA-1, 2, & 3
Ovex EPA-1
Captafol EPA-1
Carbofuran EPA-1
Carboxin EPA-1
MH EPA-1
Bensulide EPA-1
-------
14
S-(0,0-diisopropyl phosphorodithioate)
ester of N-(2-mercaptoethyl
benzenesulfonamide
N-2-(Q>, 0-diisopropyl-phosphorothiolo-
thionyl)ethyl benzenesulfonamide
0, 0-dimethyl dithiophosphate of
diethyl mercaptosuccinate
0, 0-dimethyl-0-(2-methylcarbamoyl-
1 - methyIvinyl)-phosphate
dimethyl-l-raethyl-2-methyl-carbamoyl-
vinyl phosphate
0, 0-dimethyl 0-p-nitrophenol
phosphorothioate
0, o-dimethyl S-(4-oxo-l,2,3-benzo-
tr iaz in-3(4H)-ylme thy1)
phosphorodithioate
cis-3-(dimethoxyphosphinyloxy)-N-
methylcrotonamide
dimethyl parathion
dimethyl phosphate of 3-hydroxy-N-
methyl-cis-crotonamide
l,l-dimethyl-3-(o',a)a'-trifluoro-m-
tolyl)urea
•
m-(3,3-dimethylureido)phenyl tert-
butylcarbamate
Dinitro
4,6-dinitro-o-cresol
2,4-dinitro-6-octylphenyl crotonate
2,6-dinitro-4-octylphenyl crotonate
Dinocap EPA-1
Dlnocap_EPA-2(tentative)
Dinoseb EPA-1 (tentative)
Bensulide EPA-1
Bensulide EPA-1
Malathion EPA-1 & 2
Monocrotophos EPA-1 & 2
Monocrotophos EPA-1 & 2
Methyl Parathion
EPA-1, 2, 3, 4, & 5
Azinphos-methyl EPA-1
Monocrotophos EPA-1 & 2
Methyl Parathion
EPA-1, 2, 3, 4, & 5
Monocrotophos EPA-1 & 2
Fluometuron EPA-1
Karbutilate EPA-1
Dinoseb EPA-1 & 2
Nitrophenols EPA-1 & 2
Dinocap EPA-1 & 2
Dinocap EPA-1 & 2
total nitrogen
IR
IR
-------
15
Dinoseb EPA-2 (tentative)
dinoseb methacrylate
dinosebe (France)
Di-on
Diphacin
diphacin (Turkey)
Diphacinone EPA-1
diphenadione
2-(d iphenylacetyl)-1,3-indand ione
Direz
disodium znethanearsonate
Disulfoton EPA-1 (tentative)
Disulfoton EPA-2 (tentative)
Disyston
Di-Syston (US)
ditranil
Dithio-systox
Diurex
Diuron EPA-1
Diuron EPA-2 (tentative)
Diuron EPA-3(tentative)
Diuron EPA-4 (tentative)
Dolmix
DN 289
DNBP
DNOC
D014
GLC-TCD
Binapacryl EPA-1
Dinoseb EPA-1 & 2
Diuron EPA-1, 2, 3, & 4
Diphacinone EPA-1
Diphacinone EPA-1
UV
Diphacinone EPA-1
Diphacinone EPA-1
Anilazine EPA-1 & 2
Arsenic Compounds EPA-3 & 4
IR
GLC-FID-IS
Disulfoton EPA-1 & 2
Disulfoton EPA-1 & 2
Dicloran EPA-1
Disulfoton EPA-1 & 2
Diuron EPA-1, 2, 3, & 4
alkaline hydrolysis & titration
HPLC
UV
IR
BHC, gamma isomer EPA-1
Dinoseb EPA-1 & 2
Dinoseb EPA-1 & 2
Nitrophenols EPA-1 & 2
Propargite EPA-1 & 2
-------
16
2,4-DP
Dormone
Dowcide 1
Dowco 132
Dowpon
Draza
Drinox
Drop-Leaf
DSMA
Dyrene
E 601
E 605
E 3314
Ectoral
El 4049
Emmatos
Embutox
Endosan
Endosulfan EPA-1
Endosulfan EPA-2 (tentative)
Endosulfan EPA-3 (tentative)
Endosulfan EPA-4 (tentative)
Endothal
endothal (Europe except Italy)
Endothall EPA-1
dichlorprop
2,4-D
o-phenylphenol
Crufomate EPA-1 & 2
Dalapon EPA-1
Methiocarb EPA-1
Heptachlor EPA-1
Sodium Chlorate EPA-1
Arsenic Compounds EPA-3 & 4
Anilazine EPA-1 & 2
Methyl Parathion
EPA-1, 2, 3, 4, & 5
Parathion EPA-1 & 2
Heptachlor EPA-1
Ronnel EPA-1 & 2
Malathion EPA-1 & 2
Malathion EPA-1 & 2
2,4-DB
Binapacryl EPA-1
alkaline hydrolysis
IR
GLC-TCD-IS
GLC-FID-IS
Endothall EPA-1 & 2
Endothall EPA-1 & 2
oxidation & titration
Endothall EPA-2 (tentative)
GLC-FID
-------
17
ephirsulfonate
Eptam
EPIC EPA-1 (tentative)
EPIC EPA-2 (tentative)
EPIC EPA-3
EPTC EPA-4 (tentative)
EPIC EPA-5 (tentative)
Eradicane
erbon
Erbon (Dow)
Esteron
Estone
Estonmite
Estron 245
Ethion EPA-1
Ethion EPA-2 (tentative)
Ethodan
ethohexadiol
Ethoprop EPA-1 (tentative)
Ethoprop EPA-2 (tentative)
Ethoprop EPA-3 (tentative)
S-ethyl cyclohexylethylthio-
carbamate
S-ethyl diisobutylthiocarbamate
0-ethyl-S,S-dipropyl phosphoro-
dithioate
S-ethyl dipropylthiocarbamate
Ovex EPA-1
EPTC EPA-1, 2, 3, 4, & 5
GLC-TCD-IS
HPLC
GLC-FID-IS
CLC-FID-IS
GLC-TCD-IS
EPTC EPA-1, 2, 3, 4, & 5
Chlorophenoxy Herbicides
EPA-1, 2, & 3
erbon
2,4-D
2,4-D
Ovex EPA-1
2,4,5-T
IR
GLC-TCD
Ethion EPA-1 & 2
Ethyl Hexanediol EPA-1 & 2
IR
GLC-TCD-IS
GLC-FID-IS
Cycloate EPA-1
Butylate EPA-1, 2, 3, 4, & 5
Ethoprop EPA-1, 2, & 3
EPTC EPA-1, 2, 3, 4, & 5
-------
18
Ethyl Hexanediol EPA-1
Ethyl Hexanediol EPA-2 ^tentative)
2-ethyl-l,3-hexanediol
ethylhexylene glycol
Ethyl Parathion
Etilon
Etrolene
Eurex
Fall
Falone
FBHC
Fence Rider
fenchlorphos (ISO and BSI)
Fermide
Femes ta
Fernimine
Fernoxone
ferroprop
Ferxone
flour sulfur
flowers of sulfur
Fluometuron EPA-1
FMC 5273
FMC 5462
FMC 9044
FMC 10242
acetylation & titration
GLC-TCD-IS
Ethyl Hexanediol EPA-1 & 2
Ethyl Hexanediol EPA-1 & 2
Parathion EPA-1 & 2
Parathion EPA-1 & 2
Ronnel EPA-1 & 2
Cycloate EPA-1, 2, & 3
Sodium Chlorate EPA-1
2,4-DEP
BHC, gamma isomer EPA-1
2,4,5-T
Ronnel EPA-1 & 2
Thiram EPA-1 & 2
2,4-D
2,4-D
2,4-D
silvex
2,4-D
Sulfur EPA-1, 2, & 3
Sulfur EPA-1, 2, & 3
IR
Piperonyl Butoxide EPA-1 & 2
Endosulfan EPA-1, 2, 3, & 4
Binapacryl EPA-1
Carbofuran EPA-1
-------
19
FMC 17370
Folcid
Folidol
Folidol M
Folpan
Folpet EPA-1
For-mal
Forron
Fortrol
Fosferno
Fosferno M50
Fosfono
Fruitone A
Fruitone T
Frumin AL
Fumarin
futnarin (Great Britain, New Zealand)
Fumazone
Furadan
Fylanon
G-24480
G-30027
G-41435
Gammexane
Gardentox
Resmethrin EPA-1, 2, 3, 4, & 5
Captafol EPA-1
Parathion EPA-1 & 2
Methyl Parathion
EPA-1, 2, 3, 4, & 5
Folpet EPA-1
IR
Malathion EPA-1 & 2
2,4,5-T
Cyanazine EPA-1
Parathion EPA-1 & 2
Methyl Parathion
EPA-1, 2, 3, 4, & 5
Parathion EPA-1 & 2
2,4,5-T
silvex
Disulfoton EPA-1 & 2
Coumafuryl EPA-1 & 2
Coumafuryl EPA-1 & 2
Dibromochloropropane EPA-1 & 2
Carbofuran EPA-1
Malathion EPA-1 & 2
Diazinon EPA-1, 2, 3, & 4
Atrazine EPA-1 & 2
Prometone EPA-1 & 2
BHC, gamma isomer EPA-1
Diazinon EPA-1, 2, 3, & 4
-------
20
Garlon
Gearphos
Gebutox
Genitox
Gesafram
Gesapon
Gesaprim
Gesarex
Gesarol
Gesatop
Gramevin
Guesarol
Gusathion
Guthion
Gyron
H 119
H 133
H 321
Haiari (British Guiana)
HCCH
HCH
Hedonal
Hedonal MCPP
Heptachlor EPA-1
1,4,5,6,7,8,8-heptachloro-3a,4,7,7a-
tetrahydro-4,7-methanoindene
silvex
Methyl Parathlon
EPA-1, 2, 3, 4, & 5
Dinoseb EPA-1 & 2
DDT EPA-1
Prometone EPA-1 & 2
DDT EPA-1
Atrazlne EPA-1 & 2
DDT EPA-1
DDT EPA-1
Slraazine EPA-1
Dalapon EPA-1
DDT EPA-1
Azinphos-methyl EPA-1
Azinphos-methyl EPA-1
DDT EPA-1
Pyrazon EPA-1
Dlchlobenll EPA-1
Methiocarb EPA-1
Rotenone EPA-3
BHC, gamma isomer EPA-1
BHC, gamma Isomer EPA-1
2,4-D
mecoprop
IR
Heptachlor EPA-1
-------
21
Heptachlorotetrahydro-4,7-methano-
indene (and related compounds)
Heptamul
Herbicide 273
Herbicide 283
Herbizole
Hercules 9573
hexachlor
hexachloran
1,2,3,4,5,6-hexachlorocyclohexane
Hexachlorohexahydromethano-2,3,4-
benzodioxathiepin-3-oxide
Hexafor
Hexathir
Hexavin
Hexyclan
Hibor
HOE 2671
HOE 2784
HOE 2810
Horraodin
Hortnotuho
Hydout
o-hydrod ipheny1
Hydrothol
6-hydroxy-3-(2H)-pyridazinone
5-(alpha-hydroxy-alpha-2-pyridyl-
benzyl) -7-(alpha-2-pyridylbenzyl-
idene-5-norborene-2,3-dicarboximide
Hyvar
Heptachlor EPA-1
Heptachlor EPA-1
Endothall EPA-1 & 2
Endothall EPA-1 & 2
Amitrole EPA-1
Terbutol EPA-1 & 2
BHC, gamma isoraer EPA-1
BHC, gamma isomer EPA-1
BHC, gamma isomer EPA-1
Endosulfan EPA-1, 2, 3, & 4
BHC, gamma isomer EPA-1
Thiram EPA-1 & 2
Carbaryl EPA-1 & 2
BHC, gamma isomer EPA-1
Bromacil EPA-1
Endosulfan EPA-1, 2, 3, & 4
Binapacryl EPA-1
Linuron EPA-1 & 2
Indolebutyric acid EPA-1
MCPA
Endothall EPA-1 & 2
o-phenylphenol
Endothall EPA-1 & 2
MH EPA-1
Norbormide EPA-1
Bromacil EPA-1
-------
22
Indolebutyric acidEPA-1
indole-3-butyric acid
3-indolebutyric acid
4-(3-indolyl)-butyric acid
Inorganic phosphorus compounds
6-12 Insect Repellent
Insectophene
Inverton 245
Iso-Comox
Isocothan
2-isovaleryl-l,3-indandione
Ixodex
jasmolins
Karathane
Karbaspray
Karbofos
Karbutilate EPA-1
Karmex
Kemate
Kiloseb
Kilprop
Kilrat
Kilsem
Kloben
KMH
Knoxweed
UV
Indolebutyric acid EPA-1
Indolebutyric acid EPA-1
Indolebutyric acid EPA-1
Phosphorus Compounds EPA-1
Ethyl Hexanediol EPA-1 & 2
Endosulfan EPA-1, 2, 3, & 4
2,4,5-T
mecoprop
Dinocap EPA-1 & 2
PMP EPA-1, 2, & 3
DDT EPA-1
Pyrethrins EPA-1
Dinocap EPA-1 & 2
Carbaryl EPA-1 & 2
Malathion EPA-1 & 2
!£
Diuron EPA-1, 2, 3, & 4
Anilazine EPA-1 & 2
Dinoseb EPA-1 & 2
mecoprop
Zinc Phosphide EPA-1 & 2
MCPA
Neburon EPA-1
MH EPA-1
EPTC EPA-1, 2, 3, 4, & 5
-------
23
Kopsol
Kop-Thiodan
Kop-thion
Korlan
Klorex
Krovar
Kuron
Kurosal
Kwik-kil
Kypfarin
Kyphos
Lanex
Lasso
Lazo
lead arsenate
lemongrass oil
Line Rider
Linurpn EJPA-1 (tentat ive )
Linuron EPA-2
Lonchocarpus
Lorox
M-74 (USSR)
MAA
Maintain 3
Malamar
Malaspray
DDT EPA-1
Endosulfan EPA-1, 2, 3, & 4
Malathion EPA-1 & 2
Ronnel EPA-1 & 2
Sodium Chlorate EPA-1
Bromacil EPA-1
silvex
silvex
Strychnine EPA-1 & 2
Warfarin EPA-1, 2, & 3
Malathion EPA-1 & 2
Fluometuron EPA-1
Alachlor EPA-1 & 2
Alachlor EPA-1 & 2
Arsenic Compounds EPA-1 & 2
Oil of Lemongrass EPA-1
2,4,5-T
HPLC
IR
Rotenone EPA-1
Linuron EPA-1 & 2
Disulfoton EPA-1 & 2
Arsenic Compounds EPA-3 & 4
MH EPA-1
Malathion EPA-1 & 2
Malathion EPA-1 & 2
-------
24
Malathion EPA-1 (tentative)
Malathion EPA-2
maleic hydrazide
Malix
Maloran
MAMA
Marlate
MB 2878
MB 3046
MCP
MCPA
MCPB
2,4-MCPB
MCPP
mecoprop
Meldane
Mephanac
Mepro
mercaptodimethur
mercaptothion
Mercuram
Merpan
Mesurol
Meta
metacetaldehyde
Metaclde
HPLC
IR
MH EPA-1
Endosulfan EPA-1, 2, 3, & 4
Chlorbromuron EPA-1
Arsenic Compounds EPA-3 & 4
Methoxychlor EPA-1 & 2
2,4-DB
MCPB
MCPA
Chlorophenoxy Herbicides
EPA-1, 2, & 3
Chlorophenoxy Herbicides
EPA-1, 2, & 3
MCPB
mecoprop
Chlorophenoxy Herbicides
EPA-1, 2, & 3
Coumaphos EPA-1, 2, & 3
MCPA
mecoprop
Methiocarb EPA-1
Malathion EPA-1 & 2
Thiram EPA-1 & 2
Captan EPA-1 & 2
Methiocarb EPA-1
Metaldehyde EPA-1, 2, 3, & 4
Metaldehyde EPA-1, 2, 3, & 4
Methyl Parathion
EPA-1, 2, 3, 4, & 5
-------
25
Metadelphene
Metaldehyde EPA-1
Metaldehydje EPA-2 (tenta tive)
Metaldehyde EPA-3 (tentative)
Metaldehyde EPA-4 (tentative)
Metaphos
metaxon
methanearsonic acid
Methiocarb EPA-1 (tentative)
Methoxone
Methoxychlor EPA-1 (tentative)
Methoxychlor EPA-2 (tentative)
2,2-bis(p-methoxyphenyl)-1,1,1-
trichloroethane 88% and related
compounds 12%
methyl-l-(butylcarbamoyl)-2-
benzimidazolecarbamate
O-methyl 0-2-chloro-4-tert-butyl-
phenol N-methylamidophosphate
A-(2-methyl-4-chlorophenoxy)
acetic acid
4-(2-methyl-4-chlorophenoxy)
butyric acid
2-(2-methyl-4-chlorophenoxy)
propionic acid
l-(2-methylcyclohexyl)-3-phenylurea
2,2'-methylenebis(4-chlorophenol)
2,2'-methylenebis(3,4,6-trichloro-
phenol)
Deet EPA-1, 2, & 3
iodinetrie titration
GLC-TCD-IS
IR
GLC-TCD
Methyl Parathion
EPA-1, 2, 3, 4, & 5
MCPA
Arsenic Compounds EPA-3 & 4
IR
mecoprop
IR
GLC-FID-IS
Methoxychlor EPA-1 & 2
Benomyl EPA-1 & 2
Crufomate EPA-1 & 2
MCPA
MCPB
mecoprop
Siduron EPA-1
Phenols & Chlorophenols
EPA-1, 3, & 8
Phenols & Chlorophenols
EPA-1, 3, & 8
Methyl Parathion EPA-1 (tentative)
HPLC
-------
26
Methyl Parathion EPA-2
Methyl Parathion EPA-3
Methyl Parathion EPA-4
Methyl Parathion EPA-5
4-(methylthio)-3,5-xylyl
N-methylcarbamate
Metiltriazotion
metmercapturon
Metobromuron EPA-1 (tentative)
Metobr^OTOuron EPA-2 (tentative)
Metobromuron EPA-3 (tentative)
Metron
MH EPA-1
MH-30
Mi Idex
2M-4KH-M
MLT
Mocap
monoammonium methanearsonate
Monocron
Monocrotophos EPA-1
Monocrotophos EPA-2
monosodium methanearsonate
Monurex
Monuron EPA-1
Monuron EPA-2
Monuron EPA-3
IR
colorimetric (visible) spectroscopy
GLC-FID-IS
GLC-FID-IS
Methiocarb EPA-1
Azinphos-methyl EPA-1
Methiocarb EPA-1
IR
GLC-FID
GLC-TCD-IS
Methyl Parathion
EPA-1, 2, 3, 4, & 5
UV
MH EPA-1
Dinocap EPA-1 & 2
MCPB
Malathion EPA-1 & 2
Ethoprop EPA-1, 2, & 3
Arsenic Compounds EPA-3 & 4
Monocrotophos EPA-1 & 2
IR
GLC-FID-IS
Arsenic Compounds EPA-3 & 4
Monuron EPA-1, 2, & 3
alkaline hydrolysis &^ jjltration
UV
IR
-------
27
Morocide
Mous-con
Mouse-tox
Moxie
MSMA
Muscatox
Nankor
1-naphthyl methylcarbatnate
neburea
Neburex
Neburon EPA-1 (tentative)
Nekos (Dutch Guiana)
Nemafume
Nemagon
Neocid
Neocidol
NIA 1240
NIA 5273
NIA 5462
NIA 9044
NIA 10242
NIA 11092
NIA 17370
Niagaratran
Nialate
Nicouline
Nivan
Binapacryl EPA-1
Zinc Phosphide
Strychnine EPA-1 & 2
Methoxychlor EPA-1 & 2
Arsenic Compounds EPA-3 & 4
Coumaphos EPA-1, 2, & 3
Ronnel EPA-1 & 2
Carbaryl EPA-1
Neburon EPA-1
Neburon EPA-1
IR
Rotenone EPA-1
Dibromochloropropane EPA-1 & 2
Dibromochloropropane EPA-1 & 2
DDT EPA-1
Diazinon EPA-1, 2, 3, & 4
Ethion EPA-1 & 2
Piperonyl Butoxide EPA-1 & 2
Endosulfan EPA-1, 2, 3, & 4
Binapacryl EPA-1
Carbofuran EPA-1
Karbutilate EPA-1
Resmethrin EPA-1, 2, 3, 4, & 5
Ovex EPA-1
Ethion EPA-1 & 2
Rotenone EPA-1
Parathion EPA-1 & 2
-------
28
Nitrophenols EPA—1
Nitrophenols EPA-2
Nitropone
Nitrox 80
Nomersan
Nor-Am
Norbormide EPA-1
Norex
NRDC 104
Nucidol
Nuvacron
Off
Oi^^f Lemongrass EPA-1 (tentative)
oil of verbena (Indian)
Omite
Organophosphorus compounds
Organqtin Compounds EPA-1
orthoboric acid
Orthocide
Orthophos
orthoxenol
OS 1898
ovatran (Argentina)
Ovex EPA-1
ovochlor
Ovotran
stannous chloride reduction
total nitrogen
Dinoseb EPA-1 & 2
Methyl Parathion
EPA-1, 2, 3, 4, & 5
Thiram EPA-1 & 2
Chloroxuron EPA-1 & 2
UV
Chloroxuron EPA-1 & 2
Resmethrin EPA-1, 2, 3, 4, & 5
Diazinon EPA-1, 2, 3, & 4
Monocrotophos EPA-1 & 2
Deet EPA-1, 2, & 3
GLC-TCD
Oil of Lemongrass EPA-1
Propargite EPA-1 & 2
Phosphorus Compounds EPA-1
oxidation, reduction, titration
Boron Compounds EPA-1
Captan EPA-1 & 2
Parathion EPA-1 & 2
o-phenylphenol
Dibromochloroprooane EPA-1 & 2
Ovex EPA-1
IR
Ovex EPA-1
Ovex EPA-1
-------
29
Outrack
7-oxabicyclo(2,2,1)heptane-
2,3-dicarboxylic acid
PCA
PCP
PDB
PDQ
Panthion
Paracide
paradichlorobenzene
Paradow
Paramar
Paraphos
Parathene
Z?Z2jyii°Jl EPA-1 (tentative)
Parathion EPA-2 (tentative)
parathion-methyl (ISO and BSI)
Parawet
Paris green
Partron M
Patoran
Pebulate EPA-1 (tentative)
Pebulate EPA-2^ ^tentative)
Peb^1 ate_ EPA-3 (tentative)
Pennamine D
Penta
Prometone EPA-1 & 2
Endothall EPA-1 & 2
Pyrazon EPA-1
pentachlorophenol
p-Dichlorobenzene EPA-1 & 2
MCPB
Parathion EPA-1 & 2
p-Dichlorobenzene EPA-1 & 2
p-Dichlorobenzene EPA-1 & 2
p-Dichlorobenzene EPA-1 & 2
Parathion EPA-1 & 2
Parathion EPA-1 & 2
Parathion EPA-1 & 2
HPLC
GLC-FID-IS
Methyl Parathion
EPA-1, 2, 3, A, & 5
Parathion EPA-1 & 2
Arsenic Compounds EPA-1 & 2
Methyl Parathion
EPA-1, 2, 3, 4, & 5
Metobromuron EPA-1, 2, & 3
GLC-TCD
GLC-FID-IS
GLC-FID-IS
2,4-D
pentachlorophenol
-------
30
Pentachlorin
pentachlorophenol
Pentaphen
p-tert-pentylphenol
Phaltan
Phenols & Chlorophenpls EPA-1
Phenols & Chlorophenols EPA-2
Phenols & Chlorophenols EPA-3
Phenol s_ &. Chlorophenols EPA-A
Phenols & Chlorophenols EPA-5
(tentative)
Phenols & Chlorophenols EPA-6
(tentative)
Phenols & Chlorophenols EPA-J
(tentative)
Phenols & Chlorophenols EPA-8
(tentative)
Phenothiazine EPA-1 (tentative)
o-phenylphenol
Phorate EPA-1
Phoskil
Phosphorus Compounds EPA-1
Phygon
PicloramEPA-1 (tentatiye)
Pindone EPA-1
Pindone EPA-2
Pindone EPA-3
DDT EPA-1
Phenols & Chlorophenols
EPA-1, 3, & 5
p-tert-amylphenol
p-tert-amylphenol
Folpet EPA-1
Definition, Structure, and
Technical Data
UV
lime fusion
bromination & titration
HPLC
GLC-TCD
GLC-TCD-FID
GLC-TCD-IS
IR
Phenols & Chlorophenols
EPA-1, 2, 4, 6, & 8
IR
Parathion EPA-1 & 2
acid digestioii and gravimetric
procedure
Dichlone EPA-1
HPLC
UV (ether extraction)
UV (pyrophosphate extraction)
UV (water-soluble Jpjrm^lati^oris
-------
31
Piperonyl Butgxide EPA-1
Piperonyl Butoxide EPA-2
Pival
pival (Portugal, Turkey)
pivaldione
2-pivalyl-l,3-indandione
Pivalyl valone
Pivalyn
PMP EPA-1
PMP EPA-2
PMP EPA-3
Poraarsol
Pramitol
precipitated sulfur
Prefar
Premerge
Primatol
Primatol A
Primatol S
Princep
Printop
prometon (ISO)
Prometone EPA-1 (tentative)
Prometone^ EPA-2 (tentative)
Propargite EPA-1 (tentative)
—»-.— . •*•- — J"T— • '•• —.— •- — •,—. . -. — - -M_ - - -
Propargite_EPA^2_ (tentatjLve)
propazine
qualitative test
GLC-FID-IS
Pindone EPA-1, 2, & 3
Pindone EPA-1, 2, & 3
Pindone EPA-1, 2, & 3
Pindone EPA-1, 2, & 3
Pindone EPA-1, 2, & 3
Pindone EPA-1, 2, & 3
UV (ether extraction)
UV (pyrophosphate extraction)
UV (water-soluble formulation)
Thiram EPA-1 & 2
Prometone EPA-1 & 2
Sulfur EPA-1, 2, & 3
Bensulide EPA-1
Dinoseb EPA-1 & 2
Prometone EPA-1 & 2
Atrazine EPA-1 & 2
Simazine EPA-1
Simazine EPA-1
Simazine EPA-1
Prometone EPA-1 & 2
GLC-TCD-IS
GLC-FID-IS
IR
GLC-TCD-IS
Chloro-Triazine Herbicides EPA-1
-------
32
prophos
S-propyl butylethylthiocarbamate
S-propyl dipropylthiocarbamate
S-propyl N,N-dipropyl thiocarbamate
Protex
Pyramin
Pyrazon EPA-1 (tentative)
Pyrethrins EPA-1
Pyrethrins EPA-2
Pyrethrins EPA-3
Quaternary Ammonium Compounds EPA-1
QuaternaryAmmonium Compounds EPA-2
Quaternary Ammonium Compounds EPA-3
Quaternary Ammonium Compounds EPA-4
Quaternary Ammonium Compounds EPA-5
Quilan
R 1582
R 1607
R 1910
R 2061
R 2063
Radapon
Ramik
Rampart
Ethoprop EPA-1, 2, & 3
Pebulate EPA-1, 2, & 3
Vernolate EPA-1, 2, & 3
Vernolate EPA-1, 2, & 3
Rotenone EPA-1
Pyrazon EPA-1
IR
Description, Structure, and
Technical Data
GLC-FID
steam distillation & titration
(Sell method)
Definition, Structure, Technical-
Data
Halogen and Nitrogen Conversion
Factors
qualitative (Auerbach) tests
ferricyanide method
Epton titration method
potentiometric titration
Benefin EPA-1 & 2
Azinphos-methyl EPA-1
Vernolate EPA-1, 2, & 3
Butylate EPA-1, 2, 3, 4, & 5
Pebulate EPA-1, 2, & 3
Cycloate EPA-1, 2, & 3
Dalapon EPA-1
Diphacinone EPA-1
Phorate EPA-1
-------
33
Ranyon
Rasikal
Raticate
RD 406
RD 4593
Reddon
Regulox
Resitox
Resmethrin EPA-1 (tentative)
Resmethrin EPA-2 (tentative)
Resmethrin EPA-3 (tentative)
Resmethrin EPA-4 (tentative)
Resmethrin EPA-5 (tentative)
Retard
Rhodiatox
Rhomene
Rhonox
rock sulfur, ground
Ro-Dec
Ro-Neet
Ronnel EPA-1
Ronnel EPA-2
Rotenone EPA-1
Royal MH-30
Ruelene
Rukseam
Ruraetan
Rutgers 6-12
Carbaryl EPA-1 & 2
Sodium Chlorate EPA-1
Norbormide EPA-1
dichlorprop
mecoprop
2,4,5-T
MH EPA-1
Coumaphos EPA-1, 2, & 3
IR
GLC-TCD
GLC-TCD-IS
HPLC
GLC-FID-IS
MH EPA-1
Parathion EPA-1 & 2
MCPA
MCPA
Sulfur EPA-1, 2, & 3
Strychnine EPA-1 & 2
Cycloate EPA-1, 2, & 3
IR
GLC-FID-IS
qualitative tests
MH EPA-1
Crufomate EPA-1 & 2
DDT EPA-1
Zinc Phosphide EPA-1 & 2
Ethyl Hexanediol EPA-1 & 2
-------
S-276
666
Salicylatiilide EPA-1
Salvo
Santochlor
Santophen 1
Santophen 20
Sappiran
Sarclex
Sarolex
SBP 1382
SD 15418
Septene
Septiphene
Seradix
Sevin
sevin (USSR)
Shed-a-Leaf
Shirlan
Shoxin
Siduron EPA-1 (tentative)
silvex
Simanex
simazine
Simazine EPA-1 (tentative)
Sinox
Disulfoton EPA-1 & 2
BHC, gamma isomer EPA-1
UV
2,4-D
p-Dichlorobenzene EPA-1 & 2
o-benzyl-p-chlorophenol
pentachlorophenol
Ovex EPA-1
Linuron EPA-1 & 2
Diazinon EPA-1, 2, 3, & 4
Resmethrin EPA-1, 2, 3, 4, & 5
Cyanazine EPA-1
Carbaryl EPA-1 & 2
o-benzyl-p-chlorophenol
Indolebutyric acid EPA-1
Carbaryl EPA-1 & 2
Carbaryl EPA-1 & 2
Sodium Chlorate EPA-1
Salicylanilide EPA-1
Norbormide EPA-1
UV
Chlorophenoxy Herbicides
EPA-1, 2, 3, & 5
Simazine EPA-1
Chloro-Triazine Herbicides EPA-1
UV
Dinoseb EPA-1 & 2
-------
35
Slo-Gro
sodium arsenate
sodium arsenite
sodium biborate
Sodium Chlorate EPA-1
sodium pyroborate
sodium tetraborate decahydrate
Solvirex
Soprathion
Soprocide
Spectracide
Spotrete
Sprout-Stop
Strathion
Str ep t omyc in _EPA_-_1.
streptomycine (France)
streptomycin hydrochloride
streptomycin nitrate
streptomycin sulfate
S trychn ine EPA-1
Strychnine EPA-2
Stuntman
sublimed sulfur
Suckerstuff
Sulfur EPA-1
Sulfur EPA-2^
Sulfur EPA-3
MH EPA-1
Arsenic Compounds EPA-1 & 2
Arsenic Compounds EPA-1 & 2
Boron Compounds EPA-1
reduction and^ ti.t_ratio_n
Boron Compounds EPA-1
Boron Compounds EPA-1
Disulfoton EPA-1 & 2
Parathion EPA-1 & 2
BHC, gamma isomer EPA-1
Diazinon EPA-1, 2, 3, & 4
Thiram EPA-1 & 2
MH EPA-1
Parathion EPA-1 & 2
UV or colorimetric spectroscope.
Streptomycin EPA-1
Streptomycin EPA-1
Streptomycin EPA-1
Streptomycin EPA-1
picric acid precipitation
UV
MH EPA-1
Sulfur EPA-1, 2, & 3
MH EPA-1
CS? extraction
barium sulfa_te_ precijpitatiori
CS^ extrac_tipn__(p_res_ence_ ace tone-
sojLubl.ejpe_st ic id es )
-------
36
Sulfur Dioxide EPA-1
sulfurous acid anhydride
sulfurous oxide
Su Sciguro Carpidor
Sutan
Synthrin
2,4,5-T
2,4,5-TB
4-2,4,5-TB
Tabatrex
Tabutrex
landex
TCC
Tekwaisa
Telvar
Temasept
Tenoran
Terbucarb
Terbutol EPA-1 (tent a tive)
Terbutol EPA-2 (tentative)
Tennil
Tersan
Tersan 1991
2,4,5,6-tetrachloro-3-
cyanobenzonitrile
iodine titration
Sulfur Dioxide EPA-1
Sulfur Dioxide EPA-1
Trifluralin EPA-1 & 2
Butylate EPA-1, 2, 3, 4, & 5
Resmethrin EPA-1, 2, 3, 4, & 5
Chlorophenoxy Herbicides
EPA-1, 2, 3, 4, & 5
Chlorophenoxy Herbicides
EPA-1, 2, & 3
2,4,5-TB
Dibutyl Succinate EPA-1
Dibutyl Succinate EPA-1
Karbutilate EPA-1
Trichlorocarbanilide EPA-1
Methyl Parathion
EPA-1, 2, 3, 4, & 5
Monuron EPA-1, 2, & 3
Brominated Salicylanilides EPA-1
Chloroxuron EPA-1 & 2
Terbutol EPA-1 6. 2
TR
GLC-FID-IS
Chlorothalonil EPA-1
Thiram EPA-1 & 2
Benomyl EPA-1 & 2
Chlorothalonil EPA-1
-------
37
2,4,5, 6-tetrachloro-l , 3-
d icyanobenzene
cis-N- [(1,1,2, 2-tetrachloroethyl) thio ]
4-cyclohexene-l , 2-dicarboximide
tetrachloroisophthalonitrile
0, 0, O1 , 0' -tetraethyl S , S ' -methylene
bisphosphorodithioate
1,2,3, 6-tetrahydro-3 , 6-dioxo-
pyridazine
tetramethylthiuram disulfide
Thifor
Thiraar
Thimet
Thimul
Thiodan
thiodemeton
thiodiphenylamine
Thionex
thiophal
thiophos
Thiram_ EPA-1
Thistrol
Thylate
Tillam
Timbo (Brazil)
timet
tin, organic compounds
TMTD
Chlorothalonil EPA-1
Captafol EPA-1
Chlorothalonil EPA-1
Ethion EPA-1 & 2
MH EPA-1
Thiratn EPA-1 & 2
Endosulfan EPA-1, 2, 3, & 4
Thirara EPA-1 & 2
Phorate EPA-1
Endosulfan EPA-1, 2, 3, & 4
Endosulfan EPA-1, 2, 3, & 4
Dlsulfoton EPA-1 & 2
Phenothiazine EPA-1
Endosulfan EPA-1, 2, 3, & 4
Folpet EPA-1
Parathion EPA-1 & 2
UV
IR
MCPB
Thiram EPA-1 & 2
Pebulate EPA-1, 2, & 3
Rotenone EPA-1
Phorate EPA-1
Organotin Compounds EPA-1
Thirara EPA-1 & 2
-------
38
tomarin
Tordon
Tormona
2,4,5-TP
Trefanocide
Treficon
Treflan
Triasyn
3,4',5-tribromosalicylanilides
Tributon
tributyltin compounds
Tricarnam
trichlorfension
1,1,1-trichloro-2,2-bis
(p-chlorophenyl)ethane
1,1,l-trichloro-2,2-bis
(p-methoxyphenyl)ethane
Trichlorocarbanilide EPA-1
3,4,4'-trichlorocarbanilide
N-trichloromethylthio-4-cyclo-
hexene-1,2-dicarboximide
N-(trichloromethylthio)phthalimide
2,4,5-trichlorophenoxy acetic acid
4-(2,4,5-trichlorophenoxy)butyrlc acid
2-(2,4,5-trichlorophenoxy)ethyl-2,2-
dichloropropionate
2-(2,4,5-trichlorophenoxy)propionic
acid
Tri-Endothal
a,a,a-trifluoro-2,6-dinitro-
N,N-dipropyl-p-toluidine
Coumafuryl EPA-1 & 2
Picloram EPA-1
2,4,5-T
silvex
Trifluralin EPA-1 & 2
Trifluralin EPA-1 & 2
Trifluralin EPA-1 & 2
Anilazine EPA-1 & 2
Brominated Salicylanilides EPA-1
2,4-D or 2,4,5-T
Organotin Compounds EPA-1
Carbaryl EPA-1 & 2
Ovex EPA-1
DDT EPA-1
Methoxychlor EPA-1 & 2
UV
Trichlorocarbanilide EPA-1
Captan EPA-1
Folpet EPA-1
2,4,5-T
2,4,5-TB
erbon
silvex
Endothall EPA-1 & 2
Trifluralin EPA-1 & 2
-------
39
Trifluralin EPA-1
Trifluralin EPA-2.
Triflurex
Trioxone
triphenyltin compounds
tris[2-(2,4-dichlorophenoxy)ethyl]
phosphite
Trolene
Tropotox
Tuads
tubatoxin
Tupersan
UC 7744
Unipon
Uniroyal
Ureabor
USR 604
Valone
Vancide
VC 9-104
Velsicol 104
Vergeraaster
Vernam
Vernojiate EPA-1
Vernola te EPA-2
Vernolate EPA-3 (tentative)
Vertron 2D
GLC-FID-IS
IR
Trifluralin EPA-1 & 2
2,4,5-T
Organotin Compounds EPA-1
2,4-DEP
Ronnel EPA-1 & 2
MCPB
Thiram EPA-1 & 2
Rotenone EPA-1
Siduron EPA-1
Carbaryl EPA-1 & 2
Dalapon EPA-1
Diclone EPA-1
Bromacil EPA-1
Dichlone 604
PMP EPA-1, 2, & 3
Thiram EPA-1 & 2
Ethoprop EPA-1, 2, & 3
Heptachlor EPA-1
2,4-D
Vernolate EPA-1, 2, & 3
IR
GLC-FID-IS
GLC-TCp-IS
2,4-D
-------
40
Viozene
Visko-Rhop
Vitavax
Vonaldehyde
Voncaptan
Vondrax
Vonduron
WARF
Warfarin EPA-1 (tentative)
Warfarin EPA-2
Warfarin EPA-3 (tentative)
Weedar
Weedazole
Weedone
Weedone 170
Weedone 2,4-DP
Weedone 2,4,5-T
Weedone 2,4,5-TP
WL 19805
Wolfatox
3Y9
Zelan
Zerdane
Zinc Phosphide EPA-1
Zinc Phosphide EPA-2
Zithiol
Zoocoumarin
Ronnel EPA-1 & 2
2,4-D
Carboxin EPA-1
MH EPA-1
Captan EPA-1 & 2
MH EPA-1
Diuron EPA-1, 2, 3, & 4
Warfarin EPA-1, 2, & 3
HPLC
UV
HPLC (sod. ium^ salt)
2,4-D or 2,4,5-T
Amitrole EPA-1
2,4-D
dichlorprop
dichlorprop
2,4,5-T
silvex
Cyanazine EPA-1
Methyl Parathion
EPA-1, 2, 3, 4, & 5
2,4-DEP
MCPA
DDT EPA-1
phosphine evolution
GLC-FPD
Malathion EPA-1 & 2
Warfarin EPA-1, 2, & 3
-------
December 1975 Alachlor EPA-1
(Tentative)
Determination of Alachlor by
Gas-Liquid Chromatography
(TCD - Internal Standard)
Alachlor is the common name for 2-chloro-2',6'-diethyl-N-(methoxy-
methyl) acetanilide, a registered herbicide having the chemical structure:
CH2— CH3
CH2—0—CH3
C— CH2—CI
II
— CH3
Molecular formula: C, .H-rtClN00
14 20 2
Molecular weight: 269.8
Melting point: 39.5 to 41.5°C
Physical state, color, and odor: odorless, cream-colored crystalline
solid (at RT)
Solubility: 242 ppm in water at 25°C; soluble in acetone, benzene,
chloroform, ethanol, ethyl acetate; slightly soluble in
heptane
Stability: hydrolyzed under strongly acid or alkaline conditions;
good resistance to decomposition by UV irradiation
Other names: Lasso (Monsanto), CP 50144, Lazo
-------
2 Alachlor EPA-1
(Tentative)
Reagents:
1. Alachlor standard of known % purity
2. Benzyl benzoate standard of known % purity
3. Chloroform, pesticide or spectro grade
4. Internal Standard solution - weigh 0.625 gram benzyl benzoate
into a 50 ml volumetric flask, dissolve in, and make to volume
with chloroform. (cone 12.5 mg benzyl benzoate/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 6' x 1/8" stainless steel, packed with 10% SE-30 on
80/100 Diatoport S (or equivalent column)
3. Precision liquid syringe: 10 pi
4. Usual laboratory glassware
Operating Conditions for TCP;
Column temperature: 225°C
Injection temperature: 235°C
Detector temperature: 235°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas flow: 25 ml/min
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Alachlor EPA-1
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.2 gram alachlor standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 10 ml of the internal
standard solution and shake to dissolve, (final cone 20 rag
alachlor and 12.5 mg benzyl benzoate/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.2 gram alachlor
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 10 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the alachlor. For
coarse or granular materials, shake mechanically for 30 minutes
or shake by hand intermittently for one hour, (final cone 20 mg
alachlor and 12.5 mg benzyl benzoate/ml)
Determination;
Inject 2 jil of standard and, if necessary, adjust the instrument
parameters and the volume injected to give a complete separation
within a reasonable time and peak heights of from 1/2 to 3/4 full
scale. The elution order is benzyl benzoate, then alachlor.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculat ion:
Measure the peak heights or areas of alachlor and benzyl benzoate
from both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-internal
standard solution as follows and calculate the average:
-------
Alachlor EPA-1
(Tentative)
I.S. - internal standard » benzyl benzoate
RF . (wt. I.S.)(% purity I^S.)(p_k._ ht. or area alachlor)
(wt. alachlor)(% purity alachlor)(pk. ht. or area I.S.)
Determine the percent alachlor for each injection of the
sample-internal standard solution as follows and calculate the
average:
% , (wt. I.S.)(% purity I.S.)(pk. ht. or area alachlor) (3b9Q)
(wt. sample) (pk. ht. or area I.S.)(RF) 'U-\\
Method submitted by Stelios Gerazounis, EPA, Region II, New York, N.Y.
Note! It has been suggested to cut down on the concentration of
internal standard, standard, and sample solutions by a factor
of 5 and increase the amount injected by a factor of 5. This
would use less standards.
-------
December 1975 Alachlor EPA-2
(Tentative)
Determination of Alachlor by
Gas-Liquid Chromatography
(FID - Internal Standard)
Alachlor is the common name for 2-chloro-2',6'-diethyl-N-(methoxy-
methyl) acetanilide, a registered herbicide having the chemical structure:
CH2
CH2 —0—CH3
Molecular formula: C,.H-C1NO-
14 20 2
Molecular weight: 269.8
Melting point: 39.5 to 41.5°C
Physical state, color, and odor: odorless, cream-colored crystalline
solid (at RT)
Solubility: 242 ppm in water at 25°C; soluble in acetone, benzene,
chloroform, ethanol, ethyl acetate; slightly soluble in
heptane
Stability: hydrolyzed under strongly acid or alkaline conditions;
good resistance to decomposition by UV irradiation
Other names: Lasso (Monsanto), CP 50144, Lazo
-------
2 Alachlor EPA-2
(Tentative)
Reagents:
1. Alachlor standard of known % purity
2. Triphenylmethane standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.15 gram triphenylmethane
into a 50 ml volumetric flask, dissolve in, and make to volume
with acetone, (cone 3 mg triphenylmethane/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm glass column packed with 5% SE-30 on 80/100
Chromosorb W HP (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration eauipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 190°C
Injection temperature: 240°C
Detector temperature: 240°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi (adjusted for specific GC)
Hydrogen pressure: 20 psi (adjusted for specific GC)
Air pressure: 30 psi (adjusted for specific GC)
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Alachlor EPA-2
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.08 gram alachlor standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 10 ml of the internal
standard solution and shake to dissolve, (final cone 8 mg alachlor
and 3 mg triphenylmethane/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.08 gram alachlor into
a small glass-stoppered flask or screw-cap bottle. Add by pipette
10 ml of the internal standard solution. Close tightly and shake
thoroughly to dissolve and extract the alachlor. For coarse or
granular materials, shake mechanically for 30 minutes or shake by
hand intermittently for one hour, (final cone 8 mg alachlor and
3 mg triphenylmethane/ml)
Determination:
Inject 1-2 fii of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is alachlor, tten triphenyl-
methane.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of alachlor and triphenyl-
methane from both the standard-internal standard solution and the
sample-internal standard solution.
-------
Alachlor EPA-2
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
I.S. » internal standard = triphenylmethane
RF = (wt' I.S._)(_% purity I.S.) (pk. ht. or area alachlor)
(wt. alachlor)(% purity alachlor)(pk. ht. or area I.S.)
Determine the percent alachlor for each injection of the sample-
internal standard solution as follows and calculate the average:
% . (wt. I.S.)(% purity I.S.)(pk. ht. or area alachlor
(wt. sample)(pk. ht. or area I.S.)(RF)
This method was submitted by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond, Virginia
23219.
Note! This method has been designated as tentative since it is a Va.
Exp. method and because some of the data has been suggested by
EPA's Beltsville Chemistry Lab. Any comments, criticisms,
suggestions, data, etc. concerning this method will be appreciated.
-------
September 1975 4-Aminopyridine EPA-1
(Tentative)
Determination of 4-Aminopyridine in
Solid Formulations by Ultraviolet Spectroscopy
4-Aminopyridine is a registered avicide and repellant having
the chemical structure:
NH2
Molecular formula: C,H,N.
.) O L
Molecular weight: 94.11
Melting point: 158°C
Physical state and color: white crystalline solid
Solubility: soluble in water, alcohol, and ether
Stability:
Other names: Avitrol (Avitrol Corp.), 4-AP
Reagents:
1. 4-aminopyridine of known % purity
2. Distilled water
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Filtration apparatus
4. Usual laboratory glassware
-------
4-Aminopyridine EPA-1
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.06 gram 4-aminopyridine standard into a 100 ml
volumetric flask. Dissolve, make to volume with distilled water,
and mix thoroughly. Pipette a 10 ml aliquot into a 200 ml
volumetric flask and make to volume with wateui Mix thoroughly
and pipette a 10 ml aliquot into a 100 ml volumetric flask. Make
to volume and again mix thoroughly, (final cone 3
Preparation of Sample;
Weigh a portion of sample equivalent to 0.003 gram of
4-aminopyridine into a 300 ml Erlenmeyer glass-stoppered flask.
Add 100 ml distilled water by pipette and shake on a mechanical
shaker for one hour. Filter and pipette 10 ml of the clear
filtrate into a 100 ml volumetric flask. Make to volume with
distilled water and mix thoroughly, (final cone 3 tig 4-amino-
pyridine /ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
363
balance the pen for 0 and 100% transmission at 30£ nm with dis-
tilled water in each cell. Scan both the standard and sample
from 300 nm to 210 nm with distilled water in the reference cell.
Measure the absorbance of both standard and sample at 302 nm.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent 4-aminopyridine as follows:
2 _ (abs. sample)(cone. std in ug/ml)(% purity std)
(abs. std)(cone, sample in ug/ml)
Method submitted by Stellos Gerazounis, EPA Product Analysis Lab,
Region II, New York, N. Y.
-------
October 1975
Amitrole EPA-1
Determination of Amitrole by
Visible (Colorimetric) Spectroscopy
Amitrole is the accepted common name for 3-araino-lH-l,2,4-triazole,
a registered herbicide having the chemical structure:
H
N-
•C—NH2
Molecular formula: C.H.N.
244
Molecular weight:
Melting point:
84.1
159°C
Physical state, color, and odor: white crystalline powder; odorless
when pure; bitter taste
Solubility: soluble in water (28 g/100 ml at 25°C); insoluble in
non-polar solvents, acetone, ethyl ether, oils, carbon
tetrachloride
Stability: reacts with most acids and bases to form salts, oxidizes
to azotriazole; forms derivatives with aldehydes and
ketones; strong chelating agent; somewhat corrosive to
iron, aluminum, and copper
Other names: aminotriazole (France, Great Britain, New Zealand, USSR),
3-amino-s-triazole, Amerol, Amizol, ATA, Cytrol, Herbizole,
Weedazol
-------
2 Amitrole EPA-1
Reagents;
1. Amitrole standard of known % purity
2. Sodium nitroferricyanide solution - weigh 5.96 grams
Na2Fe(CN)5N0.2H 0 into a 100 ml volumetric flask;
dissolve and make to volume with water.
3. Potassium ferrocyanide solution - weigh 8.44 grams
K4Fe(CN)6.3H20 into a 100 ml volumetric flask;
dissolve and make to volume with water.
4. Sodium hydroxide solution, 10% w/v - dissolve 10 grams
of NaOH in water and make to 100 ml.
5. Hydrogen peroxide solution 3% - dilute 30% solution 1:10.
6. Glacial acetic acid
7. Color reagent - mix 20 ml sodium nitroferricyanide solution,
20 ml potassium ferrocyanide solution, 10 ml sodium
hydroxide solution, 50 ml hydrogen peroxide solution,
and add 1.2 ml glacial acetic acid. This solution should
not be mixed until needed because it is not stable for
more than one hour.
Equipment;
1. UV-visible spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Usual laboratory glassware
Procedure;
Preparation of Standard:
Weigh 0.1 gram amitrole standard into a one liter volumetric
flask; dissolve and make to volume with water, (cone 100
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3 Amitrole EPA-1
Preparation of Sample:
Weigh a portion of sample equivalent to 0.1 gram amitrole
into a one liter volumetric flask; dissolve and make to volume
with water. (cone 100 ug amitrole/ml)
Color Formation;
Pipette 25 ml of standard into a 100 ml volumetric flask
and dilute to about 70 ml with water. Pipette 25 ml of sample
in a second flask, and,for a reagent blank, add 70 ml water to
a third flask.
To each of the three flasks, add 0.15 ml of 10% sodium
hydroxide solution and 10 ml of the color reagent. Make to
volume with water, mix well, and allow to stand at room temp-
erature for two hours. Filter if necessary to obtain clear
solutions.
Spectrophotometric Determination;
With the spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 634 nm with the
reagent blank in each cell. Scan both the standard and sample
from 750 nm to 550 nm with the reagent blank in the reference
cell.
Measure the absorbance of both standard and sample at 634 nm.
(Amitrole gives a deep green color which appears gradually.
The blank is yellow, but absorbs very little at 634 nm. Beer's
law is followed.)
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent amitrole as follows:
7 = (abs. sample)(cone, std in >ig/ml) (% purity std)
(abs. std)(cone, sample in ug/ml)
-------
October 1975
AMS EPA-1
Determination of AMS
by Sodium Nitrite Titration
AMS is the common name for ammonium sulfamate, a registered
herbicide having the chemical structure:
0
H2N S — 0 —NH4
Molecular formula: H,N00,S
b / J
Molecular weight:
Melting point:
114.1
130°C, decomposing at 160°C; the technical
product is at least 97% pure and has a m.p. of
131 to 132°C.
Physical state, color, and odor: Colorless, odorless, crystalline
solid (forms plates)
Solubility: 216 g/100 g water at 25°C; soluble in glycerol, glycols,
and formamide; hygroscopic
Stability: decomposed by heat to non-flammable gases and hence
has flame retardant properties; readily oxidized by
bromine and chlorine; forms additional products with
aldehydes; somewhat corrosive to mild steel and some
other metals
Other names: Animate (DuPont), Amcide (Albright and Wilson Ltd),
Ammonium sulfamate
-------
2 AMS EPA-1
Reagents:
1. AMS of known % purity
2. Sodium nitrite, 0.2 N solution - dissolve 2.3 grams reagent
grade sodium nitrite in water and dilute to 500 ml.
Standardize against ammonium sulfamate using the same
procedure as for the sample determination.
3. Starch iodide paper - impregnate strip of filter paper with
a freshly prepared solution of 10 grams starch and 1 gram
potassium iodide in 200 ml boiling water. Dry and store
in airtight jars or bottles.
4. Sulfuric acid, 10% solution
Equipment:
1. Titration apparatus
2. Usual laboratory glassware
Procedure:
Weigh a portion of sample equivalent to 0.2 gram of AMS into a
300 ml glass-stoppered Erlenmeyer flask; add 100 ml distilled water
and 10 ml 10% sulfuric acid. Titrate slowly with standard 0.2 N
sodium nitrite solution. Shake flask vigorously after each addition
of nitrite solution to aid in the removal of the nitrogen which is
evolved. Near the end point, the titration must be done drop by drop
with shaking after each addition.
The end point is determined by dipping a glass rod into the
solution being titrated and touching it quickly to a piece of starch-
iodide paper. An intense blue-black color must appear immediately
and must be obtained repeatedly during a 1-minute period without
further addition of nitrite solution.
-------
AMS EPA-1
Calculation:
(ml NaNO )(N NaNO )(.03803)(100)
% AMS - — =-?
(grams sample)
The milliequivalent weight of sodium nitrite for this determina-
tion is 0.0230.
(69.01) _ r
(3)(1000)
Reactions;
NH,SO,NH^ + NaNO,
2NaN02 -t- 2KI + AHC1 ^ I2 + 2KC1 + 2NaCl -f 2NO +
-f starch blue-black color
This method was adopted for use from "Comparison of Methods for
Determination of Sulfamates," W.W. Bowler and E.A. Arnold, Anal.
Chem. 19, 336 (1947) by Stelious Gerazounis, Chemist, PAL Region
II, New York
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October 1975
Anilazine EPA-1
(Tentative)
Determination of Anilazine
by Infrared Spectroscopy
Anilazine is the common name for 2,4-dichloro-6-(o-chloroanilino)-
s-triazine, a registered fungicide having the chemical structure:
Molecular formula: C.H.C1-N.
95 34
Molecular weight: 275.5
Melting point: 159 to 160°C
Physical state and color: white to tan crystalline solid
Solubility: practically insoluble in water; soluble in hydrocarbons
and most organic solvents
Stability: stable in neutral or slightly acid media; hydrolyzed by
alkali on heating; compatible with most other pesticides
Other names: Dyrene (Chemagro); B-622 (Ethyl Corp.); Direz; Kemate;
Triasyn; 4,6-dichloro-N-(2-chlorophenyl)-l,3,5-triazin-
2-amine
Reagents;
1. Anilazine standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
Anilazine EPA-1
(Tentative)
Equipment;
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.5 mm NaCl or KBr cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.05 gram anilazine standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml chloroform by
pipette, close tightly, and shake to dissolve. Add a small
amount of anhydrous sodium sulfate to insure dryness. (final
cone 5 mg/ml)
Preparation of Sample:
Weigh an amount of sample equivalent to 0.5 gram anilazine
into a glass-stoppered flask or screw-cap tube. Add 100 ml
chloroform by pipette and 1-2 grams anhydrous sodium sulfate.
Close tightly and shake for one hour. Allow to settle; centrifuge
or filter if necessary, taking precautions to prevent evaporation.
(final cone 5 mg anilazine/ml)
Determination:
With chloroform in the reference cell, and using the optimum
quantitative analytical settings, scan both the standard and
sample from 1480 cm~ to 1275 cm" (6.75 ji to 7.85 ji) .
Determine the absorbance of standard and sample using the peak
at 1375 cm (7.27 ji) and basepoint 1333 cm"1 (7.5 /a).
-------
Anilazine EPA-1
(Tentative)
Calculation;
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent anilazine as
follows:
7 - (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method submitted by Eva Santos, EPA, Region IX, San Francisco,
California.
-------
November 1975
Determination of Anilazine
by Gas—Liquid Chromatography
(TCD - Internal Standard)
Anilazine EPA-2
(Tentative)
Anilazine is the common name for 2,4-dichloro-6-(o-chloroanilino)-
s-triazine, a registered fungicide having the chemical structure:
Molecular formula: CftH_Cl0N.
95 34
Molecular weight: .275.5
Melting point: 159 to 160°C
Physical state and color: white to tan crystalline solid
Solubility: practically insoluble in water; soluble in hydrocarbons
and most organic solvents
Stability: stable in neutral or slightly acid media; hydrolyzed by
alkali on heating; compatible with most other pesticides
Other names: Dyrene (Chemagro); B-622 (Ethyl Corp.); Direz; Kemate;
Triasyn; 4,6-dichloro-N-(2-chlorophenyl)-l,3,5-triazin-
2-amine
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2 Anilazine EPA-2
(Tentative)
Reagents:
1. Anilazine standard of known % purity
2. Dieldrin standard of known HEOD content
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh an amount of dieldrin
equivalent to 0.25 gram HEOD into a 25 ml volumetric flask,
dissolve in, and make to volume with acetone, (cone 10 mg
HEOD/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. 6' x 1/8" SS column packed with 10% SE-30 on 80/100 mesh
Diatoport S (or equivalent column)
3. Precision liquid syringe - 50 ul
4. Usual laboratory glassware
Operating Conditions for TCD;
Column temperature: 215°C
Injection temperature: 230°C
Detector temperature: 230°C
Filament current: 200 ma
Carrier gas: Helium
Flow rate: 30 ml/min
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Anilazine EPA-2
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.08 gram anilazine standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 10 ml of the internal
standard solution and shake to dissolve, (final cone 8 rag
anilazine and 10 mg HEOD/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.08 gram anilazine
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 10 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the anilazine. For
coarse or granular materials, shake mechanically for 10-15
minutes or shake by hand intermittently for 25-30 minutes. Filter
if needed, (final cone 8 mg anilazine and 10 mg HEOD/ml)
Determination:
Inject 25-50 pi of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is anilazine, then HEOD.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of anilazine and HEOD from
both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
-------
Anilazine EPA-2
(Tentative)
m (wt. HEOD)(% purity HEOD)(pk. ht. or area anilazine)
(wt. anilazine)(% purity anilazine)(pk. ht. or area HEOD)
Determine the percent anilazine for each injection of the
sample-internal standard solution as follows and calculate the
average :
v , . HEOD)(% purity HEOD)(pk. ht. or area anilazine) (-tee)
* (wt. sample) (pk. ht . or area HEOD) (RF) c<~ /)
Method contributed by Arthur 0. Schlosser, EPA, Region II, New York, N. Y.
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January 1976 Arsenic Compounds EPA-1
Determination of Sodium Arsenite and
Sodium Arsenate in Aqueous Formulations
Sodium arsenite and sodium ar"senate have been registered for
pesticide use both as insecticides and herbicides. These uses have
been superseded or discontinued because of the hazard to man and
animals.
Sodium arsenate;
Sodium arsenate, dibasic or disodium hydrogen arsenate -
molecular formula Na?H AsO,; molecular weight 185.91; very soluble in
water, slightly soluble in alcohol; forms heptahydrate (7H20)t
an odorless, crystalline solid that effloresces in warm air, loses
water and becomes anhydrous at 100°C, forms pyroarsenate at 150°C or
higher. POISONOUS!
Sodium arsenite:
Molecular formula approx. NaAsCL; molecular weight 129.90; white
or grayish-white powder; somewhat hygroscopic; absorbs CO- from air;
freely aoluble in water, slightly in alcohol; VERY POISONOUS!
Principle of the Method;
4
Arsenic in aqueous formulations containing no other oxidizable or
reducible substances may be titrated directly with iodine (for arsenite)
or indirectly with thiosulfate (for arsenate) without any special sample
treatment.
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2 Arsenic Compounds EPA-1
The reaction:
As03~ + I2 + H20 ^=^t AsO^ + 2I~ + 2H+
may be made to go to completion in either direction; therefore, either
arsenate, arsenite, or both in the same solution can be determined.
Sodium arsenite may be titrated directly with iodine in a neutral
solution (acid solution plus excess sodium bicarbonate) as in the
reaction:
+ I2 + 3NaHCO — > Na2H AsC>4 + 2NaI
Sodium arsenate may be titrated indirectly using thiosulfate to
titrate the equivalent iodine liberated from KI in acid solution
(fairly concentrated hydrochloric acid) as in the reactions:
+ 2HI
203 - $ Na2S4°6
Reagents :
1. Iodine, 0.05N standard solution
2. Sodium thiosulfate, 0.05N standard solution
3. Concentrated hydrochloric acid, ACS
4. Dilute hydrochloric acid
5. Sodium bicarbonate, ACS
6. Starch indicator solution
7. Potassium iodide, crystals, ACS
8. Distilled water, boiled and cooled to remove dissolved oxygen
9. Concentrated sulfuric acid, ACS
-------
Arsenic Compounds EPA-1
Equipment;
1. Titration apparatus
2. Hot plate
3. Usual laboratory glassware
Procedure:
Determination of Sodium Arsenite;
Weigh a portion of sample equivalent to 0.05 gram arsenic
(0.087 gram NaAsO ) into a 500 ml iodine flask, dilute with water
to about 200 ml, add a few drops phenolphthalein, and acidify with
dilute hydrochloric acid, adding an excess of 2-3 drops.
Neutralize with sodium bicarbonate (in small amounts to prevent
excessive foaming) and add 4-5 grams in excess. Add 5 ml starch
indicator solution and titrate with standard iodine solution to
the first permanent blue color.
Correct for the quantity of iodine solution necessary to
produce the same color using the same reagents in the same quan-
tities as above. From the ml iodine used, calculate the percent
sodium arsenite in the sample as follows:
arsenic
(ml iodine)(N iodine)(.03746)(100)
(grams sample)
(milliequivalent weight arsenic - 0.03746)
% sodium arsenite = % arsenic X 1.734
If the arsenite results are lower than expected, another portion
of sample should be checked using the reduction procedure as under
"Determination of total arsenic: Method B" below.
-------
Arsenic Compounds EPA-1
Determination of Sodium Arsenate:
Weigh a portion of sample equivalent to 0.05 gram arsenic
(0.124 gram Na H AsO ) into a 500 ml iodine flask, dilute with
water to about 200 ml, add 5 grams potassium iodide, and shake
until dissolved. Add 2 grams sodium carbonate, shake to dissolve,
and add 7-8 ml concentrated hydrochloric acid. Cover and set in
the dark for 5-10 minutes to allow completion of the reaction.
Titrate with 0.05N sodium thiosulfate solution. When the
iodine color becomes faint, add 5 ml starch indicator solution
and titrate until the blue starch-iodine color just disappears.
Calculate the percent sodium arsenate as follows:
„ . (ml thiosulfate)(N thiosulfate)(.03746)(100)
7o arsenic = -y :—r
(grams sample)
(milliequivalent weight arsenic = 0.03746)
% sodium arsenate = % arsenic X 2.481
Determination of Total Arsenic (Arsenate + Arsenite);
Method A - Using a portion of sample equivalent to 0.05 gram
arsenic, titrate the arsenite arsenic as above under determination
of sodium arsenite. Calculate as percent arsenic and as percent
sodium arsenite.
Adjust conditions and titrate the arsenate arsenic as above
under determination of sodium arsenate. Calculate as percent
total arsenic.
Subtract the percent arsenic obtained in the arsenite procedure
from the percent total arsenic to get the percent arsenate arsenic.
Calculate this as percent sodium arsenate.
Method B - Using a portion of sample equivalent to 0.05 gram of
arsenic, reduce all the arsenic to arsenite as follows: make to
about 100 ml volume with water, add 3 ml sulfuric acid and one gram
-------
5 Arsenic Compounds EPA-1
potassium iodide, and boil until volume is approximately 40 ml.
Cool, dilute to 200 ml, and add sodium thiosulfate solution drop-
wise until the iodine color just disappears (do not use starch
indicator at this point). Neutralize with sodium bicarbonate
and add 4-5 grams excess. Add 5 ml starch indicator solution and
titrate with standard iodine solution to the first permanent blue
color. Calculate the percent total arsenic as follows:
% total arsenic = (ml iodine)
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January 1976 Arsenic Compounds EPA-2
Determination of Inorganic Arsenic Compounds in Formulations
by Digestion, Reduction, and Titration
Inorganic arsenic compounds have been registered for pesticide
use. Examples include the following:
insecticides - copper acetoarsenate (Paris green), lead arsenate,
calcium arsenate and arsenite, sodium arsenate
(ant syrups)
herbicides - sodium arsenite, arsenic acid
rodenticides - arsenic trioxide
Some of these uses have been superseded or discontinued because of
the hazard to man and animals.
Arsenic is a silver gray or tin-white brittle, crystalline metal
that turns black in air: atomic symbol, As; atomic weight, 74.92;
m.p. 818°C at 36 atm.; sublimes at 760 mm at 615° without melting;
insoluble in water; not attacked by cold H^SO, or HC1; converted by
HNO. or hot H.SO, into arsenous or arsenic acid; forms inorganic and
organic compounds, valence numbers: -3, +3, and +5
This method is primarily for sodium arsenite or sodium arsenate in
ant bait syrups. For inorganic arsenicals containing calcium, copper,
lead, etc., refer to the methods of the AOAC.
Principle of the Method;
A portion of sample is digested with concentrated nitric and
sulfuric acids; the resulting arsenate is reduced to arsenite and titrated
with standard iodine in neutral solution. Other compounds reducible or
oxidizable by iodine will interfere.
-------
2 Arsenic Compounds EPA-2
Reagents;
1. Concentrated sulfuric acid, ACS
2. Concentrated nitric acid, ACS
3. Fuming nitric acid, ACS
4. Potassium iodide, crystals, ACS
5. Sodium thiosulfate solution, 0.05N (approx.)
6. Sodium bicarbonate, powder, ACS
7. Iodine, 0.05N standard solution
8. Starch indicator solution
Equipment:
1. 500 ml Kjeldahl flask
2. Digestion apparatus: Meker burner, asbestos board with a
1.5-2 inch diameter hole, fume hood
3. Hot plate
4. Titration apparatus
5. Usual laboratory glassware
Procedure;
Digestion:
Weigh a portion of sample equivalent to 0.05 gram arsenic and
transfer to a 500 ml Kjeldahl flask (avoid getting any sample on
the neck of the flask). Cautiously add 6-8 ml concentrated sulfuric
acid and 2 ml concentrated nitric acid. Heat over a low flame until
the mixture begins to darken; then add a few drops of fuming nitric
acid (or a few ml of concentrated nitric acid). Continue heating
(adding a little nitric acid when mixture darkens) until all the
organic matter is destroyed (solution no longer darkens). Continue
heating to dense white fumes of sulfur trioxide.
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3 Arsenic Compounds EPA-2
Cool, add 15-20 ml water, pouring down the side of the flask, and
heat to fumes of sulfur trioxide (to decompose any nitrosylsulfuric
acid). Repeat with two more additions of 10-15 ml of water until
all the nitric oxide fumes are expelled. Cool.
Reduction:
Transfer the contents of the Kjeldahl flask to a 500 ml Erlen-
meyer flask and dilute with water to about 100 ml. Add one gram
potassium iodide, heat to boiling, and boil until a pale straw
color develops, but do not go below 40 ml. If heating is continued
too long after the proper color is reached, the solution will
darken and the analysis is ruined. Cool, dilute to 150-200 ml, and
remove excess free iodine by adding approx. 0.05N thiosulfate solu-
tion dropwise until the iodine color is gone. Starch indicator
should be avoided; however, if the solution is slightly colored
from organic matter or other cause than free iodine, it may be
necessary to use a few drops at this time.
Titration;
Neutralize the solution with sodium bicarbonate added in small
portions to prevent excessive foaming, and then add 4-5 grams excess.
Add 5 ml starch indicator solution and titrate with 0.05N standard
iodine solution to the first permanent blue color.
Calculation;
Calculate the percent arsenic as follows:
„ . (ml iodine)(N iodine)(0.03746)(100)
/> arsenic = -, -.—*
(grams sample)
milliequivalent arsenic » 0.03746
% sodium arsenite = % arsenic X 1.734
% sodium arsenate « % arsenic X 2.481
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January 1976 Arsenic Compounds EPA-3
(Tentative)
Determination of Organic Arsenic Compounds in Formulations
by Digestion, Reduction, and Titration
Organic arsenic compounds have been registered for pesticide use.
Examples include the following: ammonium methanearsonate (Ansar) ,
cacodylic acid, disodium methanearsonate (DSMA) , monoammonium methane-
arsonate (MAMA) , monosodium methanearsonate (MSMA) . All of these
compounds are herbicides.
The following data on methanearsonic acid (IIAA) will give an
idea of the general characteristics of this group of compounds:
n ^
Structural formula: II S
CH-7 — As<
\
XOH
Molecular formula: CH AsO,,
Molecular weight: 140.0
Physical state, color, and odor: odorless, white, crystalline solid
Melting point: 161°C
Solubility: very soluble in water and alcohol
Stability: nonflammable; mildly corrosive; stable on storage,
although solid formulations are somewhat hygroscopic;
calcium, magnesium, and iron tend to precipitate the
water-insoluble methanearsonate salts of these ions
This method is intended particularly for formulations of disodium
methanearsonate. It is not applicable in the presence of iron, copper,
chromium, manganese, tin, etc. Also, it should not be used on entirely
inorganic compounds such as arsenates or arsenites, although the addition
of sucrose (0.1 gram) is supposed to make the method reliable.
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2 Arsenic Compounds EPA-3
(Tentative)
Principle of the Method:
A portion of sample is digested for a definite period of time with
concentrated sulfuric acid and fuming nitric acid in a Kjeldahl flask
fitted with a cold finger. The arsenic is then reduced to arsenite by
potassium iodide and titrated by standard iodine solution in the neutra-
lized sample solution.
Reagents:
1. Concentrated sulfuric acid, ACS
2. Fuming nitric acid, ACS
3. Ammonium sulfamate, ACS
4. Potassium iodide, ACS, 10% solution
5. Sodium thiosulfate solution, approx. 0.1N
6. Sodium carbonate, ACS, approx. 4N (212 g/1)
7. Sodium bicarbonate, ACS, powder
8. Iodine, 0.1N standard solution
Equipment:
1. 500 ml Kjeldahl flask, fitted with a cold finger
2. Digestion apparatus: Meker burner, asbestos board with 1.5-2
inch diameter hole, stand to hold flask one inch above burner
surface, fume hood
3. Titration apparatus
4. Usual laboratory glassware
-------
Arsenic Compounds EPA-3
(Tentative)
Procedure:
Digestion;
Weigh a portion of sample equivalent to 0.08-0.10 gram arsenic
and transfer to a 500 ml Kjeldahl flask, taking care that none
adheres to the neck of the flask. Add 5.5 ml concentrated sulfuric
acid and swirl gently to dissolve or to thoroughly wet the sample.
Add 1-2 ml fuming nitric acid and place the flask on the digestion
rack with the cold finger in place. Adjust so that the flask is
one inch above the surface of the burner and digest for 55 minutes.
There will be copious evolution of nitrogen oxide fumes which will
escape past the cold finger. If evolution of these fumes ceases
before the end of the digestion period, cautiously add a few more
drops of nitric acid. After the 55 minute digestion, remove the
cold finger and continue digestion to white fumes.
Remove the flask from the burner and let cool about 5 minutes.
(The amount of cooling is best determined by experience. The flask
and contents should cool just to the extent that the contents do
not spatter badly when additional reagents are added.) Add 1.5
grams ammonium sulfate through a funnel so that it drops directly
into the bottom of the flask. Mix vigorously for one minute, then
cool under cold tap water.
Reduction:
Add 60 ml water and 10 ml potassium iodide solution and replace
the flask on the burner with the cold finger in place. Boil until
the solution is straw-colored from the iodine vapor which is evolved.
(Do not boil after the proper color is reached or the solution will
darken and the experiment is ruined because of the decomposition
products formed.)
Remove the flask from the heat and add approx. 0.1N thiosulfate
solution dropwise until the excess free iodine is gone as shown by
the solution becoming colorless. Immediately add 70 ml water, mix
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4 Arsenic Compounds EPA-3
(Tentative)
well, and carefully pour the solution into 50 ml of the sodium
carbonate solution contained in a 500 ml Erlenmeyer flask. This
should be done slowly to avoid loss of solutions caused by too
vigorous an evolution of carbon dioxide. Rinse the Kjeldahl flask
thoroughly, adding the washings to the Erlenmeyer flask.
Titration:
Complete the neutralization of the acid sample solution with
sodium bicarbonate and add a slight excess. Add 5 ml starch
solution and titrate with the 0.1N standard iodine solution to
the first permanent blue color.
Calculation:
Calculate the percent arsenic as follows:
„ . _ (ml iodine)(N iodine)(0.03746)(100)
7o arsenic — ^ i %
(grams sample)
milliequivalent weight arsenic = 0.03746
Calculate the percent organic compound by multiplying the per-
cent arsenic by the factor arsenic to compound.
Example: for disodium methanearsonate (40.74% arsenic)
% = % arsenic X 2.455
This method is essentially that of the Vineland Chemical Co., Vineland,
New Jersey.
The "tentative" designation has been placed on this method because
reports from State and EPA chemists show: (1) some never use it,
(2) some found it unsatisfactory, (3) some use it and find it satisfactory.
Also, the method is originally for formulation of disodium methanearsonate,
but it is suggested for all similar organic compounds. Any criticisms,
suggestions, additions, deletions, or data are welcome.
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February 1976 Arsenic Compounds EPA-4
Determination of Arsenic in Organic Compounds
by Sulfuric Acid Digestion and Iodine Titration
For information on organic arsenic compounds of the type for
which this method is suitable, see Arsenic Compounds EPA-3.
Principle of the Method:
A portion of sample is digested with sulfuric acid in the presence
of some organic material either inherent in the sample or added (e.g.,
starch). The arsenic from the digested material is present in reduced
form, and is titrated with standard iodine in the neutralized solution.
Reagents;
1. Concentrated sulfuric acid, ACS
2. Potassium sulfate, crystals, ACS
3. Starch powder
4. Sodium hydroxide, 25% solution
5. Phenolphthalein indicator solution
6. Sodium bicarbonate, powder, ACS
7. Starch indicator solution
8. Iodine, 0.1N standard solution
Equipment:
1. Kjeldahl flask, 500 or 800 ml
2. Digestion apparatus: Maker burner, asbestos board with
1.5-2 inch diameter hole, stand, fume hood
3. Titration apparatus
4. Usual laboratory glassware
-------
2 Arsenic Compounds EPA-4
Procedure:
Digestion:
Weigh a portion of sample equivalent to 0.08-0.10 gram arsenic
and transfer to an 800 ml Kjeldahl flask, taking care that none
adheres to the neck of the flask. Add 15 grams potassium sulfate,
20 ml sulfuric acid , and about 0.3 gram of starch. Heat gently
over a low flame until the initial frothing action subsides.
Increase flame and digest at full heat for 3-4 hours or until the
solution is colorless. The flask may be lifted from the digestion
rack and swirled to dissolve any spatters of carbon adhering to
the sides not in contact with the acid.
*
There must be enough sulfuric acid in the
flask to keep the sample wet during the
digestion. In the case of large samples
or particularly those containing vermiculite,
more acid must be added at the beginning of
the digestion. Additional acid may occasion-
ally be needed during the digestion; if so,
cautiously pour 5-10 ml down the neck of the
flask and swirl gently to mix.
Neutralization and Titration;
Transfer the cooled contents of the Kjeldahl flask to a 500 ml
Erlenmeyer or iodine flask, washing the Kjeldahl flask several times
with water and adding the washings to the Erlenmeyer flask.
Add a few drops of phenolphthalein solution and neutralize the
digest mixture to just alkaline. Cool to room temperature, make
slightly acid, then neutralize with sodium bicarbonate,adding 4-5
grams excess.
Add 5 ml starch indicator solution and titrate with 0.1N
standard iodine solution to the first permanent blue color.
-------
3 Arsenic Compounds EPA-4
For most accurate results, run a blank using the same procedure
and same amounts of reagents. Subtract the blank titration from
the sample titration.
Calculation:
Calculate the percent of arsenic as follows:
„ . (ml iodine)(N iodine)(0.03746)(100)
h arsenic * ~. :—c
(grams sample;
milliequivalent weight arsenic = 0.03746
Calculate the percent organic arsenic compound by multiplying
the percent arsenic by the factor arsenic to compound.
Example: for disodium methanearsonate (40.74% arsenic)
% • % arsenic X 2.455
This method is similar to the "Arsenic in Sodium Cacodylate" method,
AOAC 12th Ed., 1975,36.044. It is a method used by the State of Florida
Pesticide Laboratory. Also it has been used for many years in the
Beltsville Chemistry Laboratory, EPA, Beltsville, Maryland.
-------
August 1975 Atrazine EPA-1
Determination of Atrazine by Infrared Spectroscopy
Atrazine is the common name for 2-chloro-4-ethylamino-6-isopropyl-
amino-l,3,5-triazine, a registered herbicide having the chemical
structure:
Cl
VI II V
CH3-CH2-N - C C - N— CH
Molecular formula: C0H, ,C1 Nc
o 14 J
Molecular weight: 215.7
Melting point: 173 to 175°C
Physical state and color: white crystalline solid
Solubility: at 27°C solubility is 33 ppm in water, 360 ppm in
n-pentane, 12,000 ppm in diethyl ether, 18,000 ppm
in methanol, 28,000 ppm in ethyl acetate, and 52,000
ppm in chloroform
Stability: stable in neutral or slightly acidic or basic media,
hydrolyzed by alkali or mineral acid at higher
temperatures
Other names: Aatrex (Ciba-Geigy Corp.), G-30027, Atranex, Gesaprim,
Primatol A
Reagents;
1. Atrazine standard of known % purity
2. Methylene chloride, pesticide or spectro grade
3. Anhydrous sodium sulfate, granular
-------
Atrazine EPA-1
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.1 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard;
Weigh 0.05 gram atrazine standard into a small glass-
stoppered flask or screw-cap bottle, add 20 ml methylene chloride
by pipette, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 2.5 mg/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.125 gram atrazine
into a glass-stoppered flask or screw-cap tube. Add 50 ml
methylene chloride by pipette and 1-2 grams anhydrous sodium
sulfate. Close tightly and shake for one hour. Allow to settle;
centrifuge or filter if necessary, taking precaution to prevent
evaporation, (final cone 2.5 rag atrazine/ml)
-------
3 Atrazine EPA-1
Determination;
With methylene chloride in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan both the standard and sample from
1755 cm"1 to 1410 cm"1 (5.7 p to 7.1 p).
Determine the absorbance of standard and sample using the
peak at 1585 cm (6.31 ji) and basepoint at 1675 cm (5.97 ji).
Calculation:
From the above absorbances and using the standard and
sample solution concentrations, calculate the percent atrazine
as follows:
„ m (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg atrazine/ml methylene chloride gives
an absorbance of approx. 0.149 in a 0.1 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
-------
October 1975 Atrazine EPA-2
(Tentative)
Determination of Atrazine by
Gas-Liquid Chromatography
(FID - Internal Standard)
Atrazine is the common name for 2-chloro-4-ethylaraino-6-
isopropylaraino-l,3,5-triazine, a registered herbicide having the
chemical structure:
Cl
? I II V
CH3—CH2—N C^ ^C N
NCH
3
Molecular formula: C-lL.Cl N
o 14 5
Molecular weight: 215.7
Melting point: 173 to 175°C
Physical state and color: white crystalline solid
Solubility: at 27°C solubility is 33 ppm in water, 360 ppm in
n-pentane, 12,000 ppm in diethyl ether, 18,000 ppm
in methanol, 28,000 ppm in ethyl acetate, and 52,000
ppm in chloroform
Stability: stable in neutral or slightly acidic or basic media,
hydrolyzed by alkali or mineral acid at higher
temperatures
Other names: Aatrex (Ciba-Geigy Corp.), G-30027, Atranex, Gesaprim,
Primatol A
-------
2 Atrazine EPA-2
(Tentative)
Reagents:
1. Atrazine standard of known % purity
2. Alachlor standard of known % purity
3. Chloroform, pesticide or spectro grade (acetone could be used)
4. Internal Standard solution - weigh 0.2 gram alachlor into a
100 ml volumetric flask, dissolve in, and make to volume
with chloroform, (cone 2 mg alachlor/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm ID glass column packed with 5% SE-30 on
80/100 mesh Chromosorb W HP (or equivalent column)
3. Precision liquid syringe: 5 or 10 jil
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 190°C
Injection temperature: 240°C
Detector temperature: 240°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi
Hydrogen pressure: 20 psi
Air pressure: 30 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response and
reproducibility.
-------
Atrazine EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.05 gram atrazine standard into a small glass-stoppered
•
flask or screw-cap tube. Add by pipette 25 ml of the internal
standard solution and shake to dissolve. (final cone 2 mg
atrazine and 2 mg alachlor/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.05 gram atrazine
into a small glass-stoppered flask or screw-cap tube. Add by
pipette 25 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the atrazine. For
coarse or granular materials, shake or tumble mechanically for
30 minutes or shake by hand intermittently for one hour.
(final cone 2 mg atrazine and 2 mg alachlor/ml)
Determination:
Inject 1-2 pi of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is atrazine, then alachlor.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of atrazine and alachlor
from both the standard-internal standard solution and the sample-
internal standard solution.
-------
Atrazine EPA-2
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. alachlor)(% purity alachlor)(pk. ht. or area atrazine)
(wt. atrazine)(% purity atrazine)(pk. ht. or area alachlor)
Determine the percent atrazine for each injection of the
sample-internal standard solution as follows and calculate the
average:
7 _ (wt. alachlor) (% purity alachlor) (pk. ht. or area atrazine)
(wt. sample) (pk. ht. or area alachlor) (RF)
This method was submitted by the Commonwealth of Virginia, Division
of Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
Note.' This method has been designated as tentative since it is a
Va. Exp. method and because some of the data has been suggested
by EPA's Beltsville Lab. Any comments, criticism, suggestion,
data, etc. concerning this method will be appreciated.
-------
September 1975
Azinphos-methyl EPA-1
Determination of Azinphos-methyl
by Infrared Spectroscopy
Azinphos-methyl is the common name for 0,0-dimethyl S-[4-oxo-l,2,3-
benzotriazin-3(4H)-y^methyl]phosphorodithioate, a registered insecti-
cide having the chemical structure:
CH3-0
\I
— S-CH2-N
CH3—<
Molecular formula: cinH12N3°3PS2
Molecular weight: 317.34
Melting point: 73 to 74°C
Physical state and color: white, crystalline solid
Solubility: about 29 ppm in water at 25°C; soluble in most organic
solvents
Stability: unstable at temperatures above 200°C; rapidly hydrolyzed
by cold alkali and acid
Other names: Guthion (Bayer), Gusathion M (Bayer), Metiltriazotion
(USSR), Carfene, Cotnion-Methyl, Bay 17147, R1582
Reagents;
1. Azinphos-methyl standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
2 Azinphos-methyl EPA-1
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.2 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
A. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, put in 1-2 grams anhydrous sodium sulfate, and
seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard;
Weigh 0.1 gram azinphos-methyl standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml carbon disulfide
by pipette, close tightly, and shake to dissolve. Add a small
amount of anhydrous sodium sulfate to insure dryness. (final
cone 10 mg/ml)
Preparation of Sample:
Weigh an amount of sample equivalent to 0.5 gram azinphos-
inethyl into a glass-stoppered flask or screw-cap tube. Add 50 ml
carbon disulfide by pipette and 1-2 grams anhydrous sodium sulfate.
Close tightly and shake for one hour. Allow to settle; centrifuge
or filter if necessary, taking precautions to prevent evaporation.
(final cone 10 mg azinphos-methyl/ml)
-------
Determination:
Azinphos-methyl EPA-1 ^^
^B^^'
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings, scan both the standard
and sample from 700 cm" to 600 cm" (14.2 u to 16.2 u).
Determine the absorbance of standard and sample using the
peak at 653.6 cm" (15.3 p) and basepoint 625 cm" (16.0/a).
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent azinphos-methyl
as follows:
2 a (abs. sample)(cone. std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 rag azinphos-methyl/ml carbon disulfide
gives an absorbance of approx. 0.033 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
Everett Greer, EPA Region IX, San Francisco, California, submitted a
similar method using:
scan range: 830 cm~ to 700 cm" (12.0 ji to 14.0 p)
analytical peak: 775.8 cm" (12.89 ji)
basepoint: 784.9 cm" (12.74 u)
cone: 12 mg/ml
-------
August 1975
Benefin EPA-1
Determination of Benefin
by Infrared Spectroscopy
Benefin is the common name for N-butyl-N-ethyl-o/,a,a-trifluoro-
2,6-dinitro-p-toluidine, a registered herbicide having the chemical
structure:
CH—CH—N—CH~CH-CH-
Molecular formula: C--Hn,F_N-0.
13 16 3 3 4
Molecular weight: 335.3
Melting point: 65 to 66.5°C
Physical state, color, and odor: Yellow-orange crystalline solid
with no appreciable odor
Solubility:
Stability:
Other names:
70 ppm in water at 25°C; readily soluble in most
organic solvents, though lower solubility in ethanol
stable, but susceptible to decomposition by ultra-
violet radiation; compatible with most pesticides
Balan (Eli Lilly), benfluralin (BS1), Balfin,
Banafin, Benalin, Binnell, Blulan, Bonalan,
Carpidor, Quilan
Reagents:
1. Benefin standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
2 Benefin EPA-1
Equipment;
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.1 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, put in 1-2 grams anhydrous sodium sulfate, and
seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.08 gram benefin standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml chloroform by pipette, and
shake to dissolve. Add a small amount of anhydrous sodium sulfate
to insure dryness. (final cone 8 mg/ml)
Preparation of Sample:
For emulsifiable concentrates (approx. 20%), weigh 2.0 grams
sample into a 50 ml volumetric flask, make to volume with chloro-
form and mix well. Add a small amount of anhydrous sodium sulfate
to insure dryness. (final cone approx. 8 mg benefin/ml)
For 2.5% granules, weigh 6.4 grams into a glass-stoppered
flask or screw-cap bottle. Add 50 ml chloroform by pipette and
1-2 grams anhydrous sodium sulfate. Close tightly and shake for
one hour. Allow to settle; centrifuge or filter if necessary,
-------
3 Benefin EPA-1
taking precaution to prevent evaporation. Evaporate a 25 ml
aliquot to less than 10 ml, transfer to a 10 ml volumetric
flask, and make to volume with chloroform. Add a small amount
of anhydrous sodium sulfate to insure dryness. (final cone
8 mg benefin/ml)
Determination:
With chloroform in the reference cell, and using the
optimum quantitative analytical settings for the particular
IR instrument being used, scan both the standard and sample
from 1400 cm"1 to 1240 cm"1 (7.1 ji to 8.1 ji) .
Determine the absorbance of standard and sample using
the peak at 1310 cm"1 (7.63 u) and a baseline from 1330 cm"1
to 1260 cm"1 (7.52 p to 7.94 /j) .
Calculation:
From the above absorbances and using the standard and
sample solution concentrations, calculate the percent benefin
as follows:
"/ = (abs. sample)(cone. std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
-------
October 1975
Benefin EPA-2
(Tentative)
Determination of Benefin by
Gas-Liquid Chromatography
(FID - Internal Standard)
Benefin is the common name for N-butyl-N-ethyl-a,a,a-trif luoro-
2,6-dinitro-p-toluidine, a registered herbicide having the chemical
structure:
CH3— CH2
Molecular formula: C10H.,FnN-0.
13 ID 3 3 4
Molecular weight: 335.3
Melting point: 65 to 66.5°C
Physical state, color, and odor: Yellow-orange crystalline solid
with no appreciable odor
Solubility: 70 ppm in water at 25°C; readily soluble in most organic
solvents, though lower solubility in ethanol
Stability: stable, but susceptible to decomposition by ultraviolet
radiation; compatible with most pesticides
Other names: Balan (Eli Lilly), benfluralin (BSI), Balfin, Banafin,
Benalin, Binnell, Blulan, Bonalan, Carpidor, Quilan
-------
2 Benefin EPA-2
(Tentative)
Reagents:
1. Benefin standard of known % purity
2. Diazinon standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.200 gram diazinon into
a 50 ml volumetric flask, dissolve in, and make to volume
with acetone. (cone 4 mg diazinon/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm ID glass column packed with 5% SE-30 on
80/100 mesh Chromosorb W HP (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 160°
Injection temperature: 210°
Detector temperature: 210°
Carrier gas: Nitrdgen
Carrier gas pressure: 60 psi
Hydrogen pressure: 20 psi
Air pressure: 30 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Benefin EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.05 gram benefin standard into a small glass-stoppered
flask or screw-cap tube. Add by pipette 20 ml of the internal
standard solution and shake to dissolve. (final cone 2.5 mg
benefin and 4 rag diazinon/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.05 gram benefin
into a small glass-stoppered flask or screw-cap tube. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the benefin. For
coarse or granular materials, shake or tumble mechanically for
30 minutes or shake by hand intermittently for one hour.
(final cone 2.5 mg benefin and 4 mg diazinon/ml)
Determination:
Inject 1-2 jil of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from
1/2 to 3/4 full scale. The elution order is benefin, then diazinon.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of benefin and diazinon from
both the standard-internal standard solution and the sample-
internal standard solution.
-------
Benefin EPA-2
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
= (wt. diazinon)(% purity diazinon)(pk. ht. or area benefin)
(wt. benefin)(% purity benefin)(pk. ht. or area diazinon)
Determine the percent benefin for each injection of the
sample-internal standard solution as follox^s and calculate the
average:
7 _ _(_wt_._ _diazinon)_(% purity diazinon) (pk. ht. or area benef in)
(wt. sample)(pk. ht. or area diazinon)(RF) /' \)
This method was submitted by the Commonwealth of Virginia, Division
of Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
Note! This method has been designated as tentative since it is a
Va. Exp. method and because some of the data has been suggested
by EPA's Beltsville Lab. Any comments, criticism, suggestion,
data, etc. concerning this method will be appreciated.
-------
August 1975
Benomyl EPA-1
Determination of Benomyl
by Infrared Spectroscopy
Benomyl is the common name for methyl l-(butylcarbamoyl)-2-
benzimidazolecarbamate, a registered fungicide having the chemical
structure: Q H
II I
C— N— C\-\2~ CH2~CH2~<
H 0
I I!
;C— N—C— 0—CH3
Molecular formula: C-.H, oN-O-
lA 18 4 3
Molecular weight: 290.3
Melting point:
decomposes without melting
Physical state, color, and odor: white crystalline solid with a
faint acrid odor
Solubility:
Stability:
Other names:
practically insoluble in water or oils, but
soluble in acetone, chloroform, or xylene
subject to decomposition in the presence of
moisture; non-corrosive to metals
Benlate (DuPont), Tersan 1991
Reagents:
1. Benomyl standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
Benomyl EPA-1
Equipment:
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.05 gram benomyl standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml chloroform by pipette, and
shake to dissolve. Add a small amount of anhydrous sodium sulfate
to insure dryness. (final cone 5 mg/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.25 gram benomyl into
a glass-stoppered flask or screw-cap tube. Add 50 ml chloroform
by pipette and 1-2 grams anhydrous sodium sulfate. Close tightly
and shake for one hour. Allow to settle; centrifuge or filter if
necessary, taking precaution to prevent evaporation. (If solution
is not clear, add a little celite, shake, and re-centrifuge or
re-filter.) (final cone 5 mg benomyl/ml)
-------
Benomyl EPA-1
Determination:
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1850 cm to
1640 cm"1 (5.4 u to 6.1 u).
Determine the absorbance of standard and sample using the
peak at 1720 cm (5.81 ji) and basepoint at 1810 cm" (5.52 p).
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent benomyl as follows:
„ = (abs. sample)(cone. std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg benomyl/ml chloroform gives
an absorbance of approx. 0.06 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Laboratory Services, 1 North 14th Street, Richmond, Virginia 23219.
-------
February 1976
Benomyl EPA-2
(Tentative)
Determination of Benomyl in Powder Formulations
by Ultraviolet Spectroscopy
Benomyl is the common name for methyl l-(butylcarbamoyl)-2-
benzimidazolecarbamate, a registered fungicide having the chemical
structure:
H
0
II
C—N— CH2-CH2-CH2-CH3
V ?
C—N—C-—0—CH3
Molecular formula: CL.H ftN 0
Molecular weight: 290.3
Melting point: decomposes without melting
Physical state, color, and odor: white crystalline solid with a faint
acrid odor
Solubility: practically insoluble in water or oils, but soluble in
acetone, chloroform, or xylene
Stability: subject to decomposition in the presence of moisture;
non-corrosive to metals
Other names: Benlate (DuPont), Tersan 1991
Reagentsi
1. Benomyl standard of known % purity
2. Dioxane, pesticide or spectro grade
-------
Benomyl EPA-2
(Tentative)
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.04 gram benomyl standard into a 50 ml volumetric
flask, dissolve, make to volume with dioxane, and mix thoroughly.
Pipette a 5 ml aliquot into a second 50 ml volumetric flask,
make to volume with dioxane, and mix thoroughly. Pipette a 5 ml
aliquot into a third 50 ml volumetric flask, make to volume with
dioxane, and again mix thoroughly. (final cone 8 ug/ml)
Allow the last solution to stand for three hours with occasional
shaking. (See note under procedure.)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.04 gram of benomyl
into a 125 ml Erlenmeyer glass-stoppered flask. Add 50 ml dioxane
by pipette and shake on a mechanical shaker for 30 minutes. Allow
to stand until a clear solution is obtained, or, if necessary,
centrifuge or filter a portion. Pipette 5 ml of the clear solution
into a 50 ml volumetric flask, make to volume with dioxane, and
mix thoroughly. Pipette 5 ml of this solution into another 50 ml
volumetric flask, make to volume with dioxane and mix thoroughly.
(final cone 8 jag benomyl/ml) Allow to stand for three hours with
occasional shaking.
-------
Benomyl EPA-2
(Tentative)
Note:
Benomyl absorbs strongly in the range of 260-310 nm.
There are three pronounced peaks when the dioxane
solution is examined immediately after the final dilution
(282 nm, 287 nm, and 294 nm). It was observed, hovever,
that the peak at 294 nm was diminishing gradually until
it practically disappeared. Since this affects the neak
at 287 nm, it is necessary to allow the solution to
stand until complete equilibrium is reached. After 3
hours of standing the absorbance at 287 nm shows no
further change. At this stage there is also a straight
relationship between concentration and absorbance.
UV Determination:
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 287 nm with
dioxane in each cell. Scan both the standard and samnle from
330 nm to 240 nm with distilled water in the reference cell.
Measure the absorbance of both standard and sample at 287 nm.
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent benomyl as follows:
y _ (abs..sample)(cone, std in pg/ml)(% purity std)
(abs. std)(cone, sample in jig/ml)
Method submitted by Stelios Gerazounis, EPA Region II, New York, N. Y.
-------
August 1975 Bensulide EPA-1
Determination of Bensulide
by Infrared Spectroscopy
Bensulide is the common name for S-(0,0-diisopropyl phosphoro-
dithioate) ester of N-(2-mercaptoethyl) benzenesulfonamide, a
registered herbicide having the chemical structure:
0 H
II i
S-N-ChUrCf-U-S-p
II 2 2
0
Molecular formula: C,/H,./NO/PS.,
14 24 4 3
Molecular weight: 397.5
Melting point: 34.4°C (supercools readily)
Physical state and color: colorless liquid or white crystalline solid
Solubility: 25 ppm in water at 20°C; slightly soluble in kerosene,
moderately soluble in xylene, and readily soluble in
acetone and raethanol
Stability: relatively stable and non-corrosive; decomposes at
elevated temperature over long periods of time (at 80°C
in 50 hr and at 200°C in 18-40 hr)
Other names: Betasan - for turf use and Prefar - for crop use (Stauffer);
N-2-(0,0-diisopropyl-phosphorothiolothionyl) ethyl benzene-
sulf onamide; diisopropyl S-(2-phenylsulfonylaminoethyl)
phosphorothiolothionate
-------
2 Bensulide EPA-1
Reagents:
1. Bensulide standard of known % purity
2. Acetone, pesticide or spectro grade
3. Carbon disulfide, pesticide or spectro grade
4. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.1 gram bensulide standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml chloroform by
pipette, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 10 mg/ml)
-------
3 Bensulide EPA-1
Preparation of Sample;
For emulsiftable concentrates, weigh a portion of sample
equivalent to 0.25 gram bensulide into a 25 ml volumetric
flask. Make to volume with chloroform and mix well. Add a
few grams anhydrous sodium sulfate to insure dryness. (final
cone 10 mg bensulide/ml)
For fertilizers, dusts, or granules, weigh a portion of
sample equivalent to 0.2 gram bensulide into a glass-stoppered
flask or screw-cap bottle. Add 50 ml chloroform and 1-2 grams
anhydrous sodium sulfate. Close tightly and shake for one hour.
Allow to settle; centrifuge or filter if necessary, taking pre-
* caution to prevent evaporation. (Virginia laboratories report
that a Soxhlet extracts too much filler from fertilizers.)
Evaporate a 25 ml aliquot of the clear solution to less than
10 ml and transfer to a 10 ml volumetric flask. Make to volume
with chloroform, mix well, and add a little anhydrous sodium
sulfate to insure dryness. (final cone 10 mg bensulide/ml)
Determination;
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and the sample from 714 cm
to 600 cm'1 (14 u to 16.5 u).
Determine the absorbance of standard and sample using the peak
at 645.2 cm (15.5 p) and a basepoint at 613.5 cm~ (16.3 p).
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent bensulide as follows;
7 = (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg bensulide/ml chloroform gives an
absorbance of approx. 0.02 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services.
-------
February 1976
BHC, gamma isomer EPA-1
Determination of BHC, gamma isomer
in Lindane Dusts by Infrared Spectroscopy
BHC is the common name for 1,2»,3,4,5,6-hexachlorocyclohexane, a
registered insecticide having the chemical structure:
Cl
.CH
CI-CH
CI-CH
SCH-CI
.CH-CI
Cl
The technical product is a mixture of five or more isomers
(65-70% alpha, 5-6% beta, 13% gamma, 6% delta — Ramsey and Patterson
(JAOAC 1946) . The insecticidal activity is due mainly to the gamma
isomer.
Lindane is the official name for a product containing not less
than 99% gamma isomer and having a melting point of not less than 112°C,
BHC, gamma isomer
Molecular formula: C,H,C1,
DO D
Molecular weight: 290.8
Melting point: 112.9°C
Physical state, color, and odor: colorless, odorless, crystals
-------
2 BHC, gamma isomer EPA-1
Solubility: 10 ppm in water at RT; slightly soluble in petroleum oils;
soluble in acetone, aromatic and chlorinated hydrocarbons
Stability: stable to air, light, heat, and carbon dioxide; unattacked
by strong acids; dehydrochlorinated by alkali
Other names: Gammexane (ICI Ltd), benzenehexachloride, HCH (Europe),
666 (Denmark), hexachlor (Sweden), hexachloran (USSR),
Benzahex, Benzex, Dolmix, FBHC, HCCH, Hexafor, Hexyclan,
Soprocide
Reagents;
1. BHC, gamma isomer of known % purity
2. Carbon disulfide, pesticide or spectre grade
3. Sodium sulfate, anhydrous, granular
Equipment;
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.5 mm cells
2. Usual laboratory glassware
Procedure:
Preparation of Standard;
Weigh 0.25 gram BHC, gamma isomer into a 50 ml glass-stoppered
flask or screw-capped bottle. Add 25 ml carbon disulfide by pipette,
shake to dissolve, and add a small amount of anhydrous sodium sulfate
to insure dryness. (cone 10 mg/ral)
-------
3 BHC, gamma isomer EPA-1
Preparation of Sample:
Weigh a portion of sample equivalent to 0.25 gram of BHC gamma
isomer into a 50 ml glass-stoppered flask or screw-capped bottle.
Add 25 ml carbon disulfide by pipette and a small amount of anhydrous
sodium sulfate; let stand for at least 30 minutes with occasional
shaking. (cone 10 mg BHC/ml)
IR Determination:
With carbon disulfide in the reference cell and the spectre-
photometer at the optimum quantitative analytical settings, scan
both the standard and sample from 770 cm to 650 cm (13 ^i to
15.4 ji). Measure the absorbance of the peak at 687 cm (14.55 p)
using a baseline from 720 cm~ to 673 cm" (13.9 p to 14.85 ji).
Calculation;
Calculate the percent of BHC, gamma isomer as follows:
j = (abs. sample)(cone, standard in mg^/ml) (% purity standard)
(abs. standard)(cone. sample in mg/ml)
-------
November 1975
Binapacryl EPA-1
(Tentative)
Determination of Binapacryl
by Infrared Spectroscopy
Binapacryl is the accepted common name for 2-sec-butyl-4,6-dinitro-
phenyl 3-methyl-2-butenoate, a registered fungicide and miticide having
the chemical structure :
CH=C— CH-*
CH-CH2-CH3
CH-
Molecular formula: C, CH N 0
lj lo 2 b
Molecular weight: 322
Melting point: 68 to 69°C
Physical state, color, and odor: white crystalline solid with faint
aromatic odor
Solubility: practically insoluble in water, but soluble in most organic
solvents
Stability: unstable in concentrated alkalis and dilute acids; slight
hydrolysis on long contact with water; slowly decomposed
by ultraviolet light: non-corrosive; compatible with W.P.
formulation of insecticides and non-alkaline fungicides
Other names: Acricid, Endosan, Morocide, HOE 2784 (Farbwerke Hoechst);
NIA 9044 (Niagara); FMC 9044; Ambox, dinoseb methacrylate
-------
2 Binapacryl EPA-1
(Tentative)
Reagents:
1. Binapacryl standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment;
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.2 mm KBr or NaCl cells
2. Mechanical shaker
3. Soxhlet extraction apparatus
4. Centrifuge or filtration apparatus
5. Rotary evaporator
6. Cotton or glass wool
7. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.10 gram binapacryl standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml chloroform by pipette, and shake
to dissolve. Add a small amount of anhydrous sodium sulfate to
insure dryness. (final cone 10 mg/ml)
Preparation of Sample:
For wettable powder or dust formulations, weigh a portion of
sample equivalent to 0.5 gram binapacryl into a glass-stoppered
flask or screw-cap bottle. Add 50 ml chloroform by pipette and 1-2
grams anhydrous sodium sulfate. Close tightly and shake for one hour,
Allow to settle; centrifuge or filter if necessary, taking precaution
to prevent evaporation. (final cone 10 mg binapacryl/ml)
-------
3 Binapacryl EPA-1
(Tentative)
If the results obtained by the above shake-out procedure on a
4% dust are low, another portion of sample should be checked using
a Soxhlet extraction as follows: Weigh an amount of sample equiv-
alent to 0.5 gram binapacryl into a Soxhlet thimble, plug with
cotton or glass wool, and extract with chloroform for 2-3 hours.
Evaporate to 30-40 ml, transfer quantitatively to a 50 ml volu-
metric flask, and make to volume with chloroform. Add a small
amount anhydrous sodium sulfate to insure dryness. (final cone
10 mg binapacryl/ml)
eraulsifiable concentrates and aqueous dispersions, weigh a
portion of sample equivalent to 0.5 gram binapacryl into a glass-
stoppered flask or screw-cap bottle. Add 50 ml chloroform by
pipette, a few boiling chips to aid agitation, and sufficient anhy-
drous sodium sulfate to absorb all the water. Close tightly and
shake vigorously on a shaking machine for one hour. Allow to settle.
Filter or centrifuge if necessary to get a clear chloroform solution,
taking precaution to prevent evaporation. (final cone 10 mg
binapacryl/ml)
Determination :
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular TR instrument
being used, scan both the standard and sample from 1540 cm to
1220 cm"1 (6.5 to 8.2 i) .
Determine the absorbance of standard and sample using the peak
346 cm"1 (7
(7.10 i to 7.85
at 1346 cm"1 (7.43 ) and baseline from 1408 cm"1 to 1273 cm"1
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent binapacryl as follows:
7 _ (abs. sample) (cone. std in ing /ml) (% purity std)
(abs. std) (cone, sample in mg/ml)
-------
February 1976 Boron Compounds EPA-1
Determination of Inorganic Boron Compounds
in Formulations by Ignition and Titration
Borax is the trivial name for sodium tetraborate decahydrate, a
registered herbicide and fungicide having the empirical formula:
Na2P^07 - 10 H20
Boric acid is a registered fungicide and insecticide having the
empirical formula:
H3B03
Borax:
Molecular formula: Na~B,07 (anhydrous)
Molecular weight: 201.3 (anhydrous)
381.4 (decahydrate)
Melting point: approx. 740°C (anhydrous)
approx. (enclosed space) 62°C (decahydrate)
Physical state, color, and odor: light, gray odorless solid (anhydrous)
white crystalline odorless solid (decahydrate)
Solubility: in 100 ml water at 20°C, approx. 2.5 g anhydrous and approx. 5 g
decahydrate; soluble in glycerol and ethylene glycol but
insoluble in ethanol
Stability: the decahydrate loses 5 molecules of water of crystallization
at 100°C, 4 more at 160°C, and becomes anhydrous at 320°C;
its aqueous solution is alkaline, but it is hydrolyzed by
mild alkali; not compatible with certain herbicides; also
used as a flame retardant and a corrosive inhibitor for
ferrous metals
Other names: sodium pyroborate, sodium biborate
-------
2 Boron Compounds EPA-1
Boric acid;
Molecular formula: H.BO
Molecular weight: 61.84
Melting point: approx. 160°C
Physical state, color, and odor: odorless, colorless crystals or white
granules or powder
Solubility: soluble in cold water, more soluble in boiling water;
soluble in alcohol or glycerol
Stability: loses one molecule of water, forming metaboric acid HBCL
when heated at 100-105°C; on long heating pyroboric acid
H«B,07 is formed, and at higher temperatures the anhydride
boric oxide BO is formed; stable in air; incompatible
with alkali carbonates and hydroxides
Other names: boracic acid, orthoboric acid
Principle of the Method:
The inorganic boron compound is extracted from the sample with
warm water. Fluorine is removed by precipitation and filtration.
Organic matter is destroyed by ignition. The boron (as boric acid) is
titrated with sodium hydroxide using mannitol as a titration aid.
Reagents:
1. Acetic acid, ACS
2. Calcium acetate, 20% solution
3. Calcium hydroxide, saturated solution
4. Hydrochloric acid, dilute
5. Methyl red indicator solution
6. Sodium hydroxide, dilute
7. Sodium hydroxide, 0.02N standard solution
8. Mannitol (see note 1)
-------
3 Boron Compounds EPA-1
Equipment:
1. Platinum dish, 150 ml
2. Muffle furnace or Meker burner
3. Filtration apparatus
4. Titration apparatus
5. Usual laboratory glassware
Procedure:
Preparation of Sample:
Weigh and transfer to a 200 ml volumetric flask a portion of
sample equivalent to 1 gram of boric acid, 1.5 grams of borax,
or 0.5 gram of boric oxide. Digest with 150 ml warm water for
15-20 minutes, shaking frequently. Cool to room temperature,
make to volume, and filter through a dry filter.
Removal of Fluorine Compounds^
Transfer a 100 ml aliquot of the filtrate to a 200 ml volumetric
flask, acidify slightly with acetic acid, and precipitate the fluorine
with an excess of calcium acetate solution. Check for complete
precipitation by allowing a few milliliters of calcium acetate solution
to run down the neck of the flask. Continue the addition of calcium
acetate until there is no evidence of additional precipitation. Make
to volume, mix thoroughly, and filter through a dry filter.
Ignition:
Pipette 100 ml of the clear filtrate into a platinum dish, add
an excess of calcium hydroxide solution, evaporate to dryness, and
ignite to destroy acetates and char other organic matter that may
be present. Avoid an intense red heat. Cool, digest with about 50 ml
hot water, and add HC1, drop by drop, until the reaction is distinctly
acid to methyl red. Filter into a 500 ml Erlenmeyer flask, washing
well with hot water.
-------
4 Boron Compounds EPA-1
Neutralization and Titration:
Exactly neutralize with sodium hydroxide; then make acid with
hydrochloric acid using an excess equivalent to 1 ml 0.2N solution.
Boil for about 5-10 minutes to expel carbon dioxide. Cool to room
temperature and neutralize with 0.2N sodium hydroxide until the
color of the solution changes from pink to yellow. If this neutral
point has been passed or if there is anv doubt, restore the pink
color with acid and bring back to yellow with the very minimum
amount of standard 0.02N sodium hydroxide.
Add 2-3 grams mannitol (note 1) and a few drops of phenol-
phthalein solution. Note the burette reading and titrate the solu-
tion with the 0.2N sodium hydroxide solution until a phenolphthalein
pink color is obtained. The addition of the mannitol causes a red
color to develop due to the presence of the methyl red indicator.
During titration of the boric acid, this color will fade. As the
titration continues, the red color due to phenolphthalein will
develop. Add a little more mannitol and if the color disappears,
continue the addition of the standard sodium hydroxide until it
again appears. Repeat until the addition of mannitol has no further
action on the end point. (note 2)
A blank should be run using the same reagents in the same
quantities as used for the sample.
Calculation:
From the volume of 0.02N sodium hydroxide solution used after
the addition of mannitol, corrected for the blank, calculate the
percent of inorganic boron compound as follows:
(ml N_aOH)(_N NaOH)(milliequivalent weight compound) (100)
= (grams sample)(100/200)(100/200)
milliequivalent weights are:
0.03482 for boric oxide B-0-
0.06184 for boric acid H-B03
0.05032 for sodium tetraborate, anhydrous Na?B,0
0.09536 for sodium tetraborate, decahydrate Na2B,07 . 10HLO
-------
5 Boron Compounds EPA-1
Notes:
(1) If mannitol is unavailable, neutral glycerol may be substituted,
using a quantity equal to one-third the volume of the solution
to be titrated, adding more if necessary.
(2) Boric acid is a weak acid in aqueous solutions and cannot be
neutralized by alkali in stoichiometric proportions. Poly-
valent alcohols such as mannitol and glycerol form complex
acids with boric acid which are much stronger than boric acid
alone and are capable of reaction with alkali.
-------
January 1976
Bromacil EPA-1
(Tentative)
Determination of Bromacil
by Gas-Liquid Chromatography
(FID - Internal Standard)
Bromacil is the accepted common name for 5-bromo-3-sec-butyl-6-
methyluracil, a registered herbicide having the chemical structure:
V
.N-
Br—C
N—CH—CH2—CH3
'C
II
0
Molecular formula: CgH-
Molecular weight: 261.1
Melting point: 158 to 159°C
Physical state, color, and odor: odorless, white crystalline solid
Solubility: 815 ppm in water at 25°C; moderately soluble in strong
aqueous bases, acetone, acetonitrile, ethanol; sparingly
soluble in hydrocarbons
Stability: temperature stable up to m.p. (gradually sublimes just
below m.p.); stable in water, aqueous bases, and organic
solvents; decomposes slowly in strong acids
Other names: Hyvar, Krovar (Du Pont); Borea, Ureabor, Borocil, Hibor
-------
2 Bromacil EPA-1
(Tentative)
Reagents:
1. Bromacil standard of known % purity
2. Dieldrin standard of known HEOD content
3. Toluene, pesticide or spectro grade
4. Internal Standard solution - weigh 0.4 gram HEOD into a 100 ml
volumetric flask; dissolve in and make to volume with toluene.
(cone 4 mg HEOD/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 4 mm ID glass, packed with a 1+1 mixture of
10% DC-200 and 15% QF-1 on 60/80 Gas Chrom Q (or
equivalent column)
3. Precision liquid syringe: 10 jal
4. Mechanical shaker
5. Centrifuge
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 200°C
Injection temperature: 220°C
Detector temperature: 300°C
Carrier gas: Nitrogen
Carrier gas pressure: 20 psi (adjusted for specific GC)
Hydrogen flow rate: adjust for specific GC
Air flow rate: adjust for specific GC
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Bromacil EPA-1
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.135 gram bromacil standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 25 ml of the internal
standard solution and shake to dissolve. (final cone 5.4 mg bromacil
and 4 mg HEOD/ml)
Preparation of Sample;
For dry formulations and oil solutions, weigh a portion of
sample equivalent to 0.135 gram bromacil into a small glass-
stoppered or screw-capped flask. Add by pipette 25 ml internal
standard solution and shake to dissolve the bromacil - at least
30 minutes for dry formulations. (final cone - see below)
For water-soluble salts and liquid formulations, weigh a
portion of sample equivalent to 0.135 gram bromacil into a 50 ml
centrifuge tube, add 0.75 ml 1+1 H?SO., and mix by swirling. Add
by pipette 25 ml internal standard solution and shake vigorously.
Centrifuge until the organic layer is clear. (final cone 5.4 mg
bromacil and 4 mg HEOD/ml)
Determination;
Inject 1-2 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2 to
3/4 full scale.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of bromacil and HEOD from both
the standard-internal standard solution and the sample-internal
standard solution.
-------
4 Bromacil EPA-1
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. HEOD)(% purity HEOD)(pk. ht. or area bromacil)
* (wt. bromacil)(% purity bromacil)(pk. ht. or area HEOD)
Determine the percent bromacil for each injection of the sample-
internal standard solution as follows and calculate the average:
./ = (wt. HEOD)(% purity HEOD)(pk. ht. or area bromacil) (
° (wt. sample) (pk. ht. or area HEOD) (RF) 'tA-
Note: The elution order was not given in the submitted method and will
have to be determined the first time this method is used.
Method submitted by Mississippi State Chemical Laboratory, Box CR,
Mississippi State, Mississippi 39762.
-------
January 1976
Brominated Salicylanilides EPA-1
Determination of Polybrominated Salicylanilides
by Ultraviolet Spectroscopy
Polybrominated salicylanilides are registered bacteriostats and
fungistats. The commercial product commonly used in formulations
contains 80% 3,4",5-tribromosalicylanilide and 20% 4',5-dibromosalicyl-
anilide and is designated as polybrominated salicylanilide. The
structure and chemical characteristics of these compounds are as follows:
3,4',5-tribromosalicylanilide
Br OH
0 H
II
C—N
Molecular formula: C,,H0Br0N00
lj O J i
Molecular weight: 449.96
Melting point: 227-228°C
Physical state, color, odor, and taste: odorless, tasteless, white powder
Solubility: insoluble in water; soluble in acetone, benzene, ethyl
acetate, ethanol, isopropanol; slightly soluble in carbon
tetrachloride
Stability: under normal temperature conditions, stable when dry and in
neutral solutions or in organic solvents; good light sensi-
-------
Brominated Salicvlanilides EPA-1
4',5-dibromosalicylanilide
0 H
C N
Molecular formula: C H Br NO
Molecular weight: 371.06
Melting point:
Physical state, color, and odor: odorless, tasteless, white powder
Solubility: (see 3,4',5-tribromosalicylanilide above)
Stability: (see 3,4',5-tribromosalicylanilide above)
Other names: Temasept
Reagents:
1. 3,4',5-tribromosalicylanilide of known % purity
2. 4',5-dibromosalicylanilide of known % purity
3. Ethanol, spectro grade
4. Sodium hydroxide, 0.1N solution
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Filtration apparatus or centrifuge
3. Usual laboratory glassware
-------
3 Brominated Salicylanilldes EPA-1
Procedure:
Preparation of Standard:
Weigh a portion of 3,4',5-tribromosalicylanilide and A',5-
dibromosalicylanilide in the ratio declared in the sample so that
the total weight is 0.1 gram (e.g., 0.08 gram tribromo- and 0.02
gram dibromo-, total 0.1 gram for 80% tribromo- and 20% dibromo-
as in the commercial polybrominated salicylanilide).
Place the weighed standard in a glass-stoppered or screw-
capped flask, add 100 ml ethanol by pipette,and shake to dissolve.
Mix thoroughly, pipette 10 ml to a 50 ml volumetric flask, and
make to volume with ethanol. Again mix thoroughly and pipette
5 ml into another 50 ml volumetric flask. Add 5 ml 0.1N sodium
hydroxide solution and 20 ml water; make to volume with ethanol
and mix thoroughly. (final cone 20 ug polybrominated salicylan-
ilide/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.02 gram polybrominated
salicylanilide into a glass-stoppered or screw-capped flask, add
100 ml ethanol by pipette, and shake to dissolve. Allow any solid
matter to settle and pipette 5 ml into a 50 ml volumetric flask.
Add 5 ml 0.1N sodium hydroxide solution and 20 ml water; make to
volume with ethanol and mix thoroughly. The solution must be clear;
if not, centrifuge or filter, taking care to prevent loss by evapora-
tion, (final cone 20 ug polybrominated salicylanilide/ml)
Prepare a blank solution using 5 ml 0.1N sodium hydroxide
solution, 20 ml water, and 25 ml ethanol. Mix thoroughly.
Determination:
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used, balance
the pen for 0 and 100% transmission at 360 nm with the blank solution
-------
Brominated Salicylanilides EPA-1
in each cell. Scan both the standard and sample from 420 nm to
320 nm with the blank solution in the reference cell. Measure
the absorbance of both standard and sample at 360 nm.
From the above absorbances and using the standard and sample
concentrations, calculate the percent polybrominated salicylanilide
as follows:
°i - ( a b s • s amp 1 e ) (cone, std in pg/ml) (% purity std)
(abs. std) (cone, sample in jig/ml)
-------
September 1975 Butylate EPA-1
(Tentative)
Determination of Butylate
by Gas-Liquid Chromatography (TCD)
Butylate is the common name for S-ethyl diisobutylthiocarbamate,
a registered herbicide having the chemical structure:
CH3
CH2 — CH — CH3
CH3— CH2 - S — C - N/
CH2 — CH - CH3
Molecular formula: C H2 ONS
Molecular weight: 217.4
Boiling point: 71°C at 10 mm
Physical state and color: Amber liquid
Solubility: 45 ppm in water at room temperature; miscible with
kerosene, acetone, methyl isobutyl ketone, ethanol, xylene
Stability: stable under ordinary conditions; non-corrosive
Other names: Sutan (Stauf fer) , R1910
Reagents:
1. Butylate standard of known % purity
2. Chloroform, pesticide or spectro grade
-------
2 Butylate EPA-1
(Tentative)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 5' x 1/4" glass column packed with 20% SE-30 on
•
Chromosorb W, AW, DMCS (or equivalent column)
3. Precision liquid syringe: 50 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for TCD;
Column temperature: 180°C
Injection temperature: 240°C
Detector temperature: 270°C
Carrier gas: Helium
Flow rate: 100 ml/min
Operating conditions for filament current, column temperature,
or gas flow should be adjusted by the analyst to obtain optimum
response and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.20 gram butylate standard into a 10 ml volumetric
flask; dissolve and make to volume with chloroform.(final
cone 20 mg/ml)
-------
Butylate EPA-1
(Tentative)
Preparation of Sample:
For technical material and liquid formulations, weigh a
portion of sample equivalent to 0.20 gram butylate into a 10 ml
volumetric flask, make to volume with chloroform,and mix
thoroughly. (final cone 20 mg butylate/ml)
For dry formulations, weigh a portion of sample equivalent
to 1.0 gram butylate into a 125 ml screw-cap flask, add by
pipette 50 ml chloroform, and shake for one hour. Allow to
settle; filter or centrifuge if necessary, taking precautions
to prevent evaporation. (final cone 20 mg butylate/ml)
Determination:
Using a precision liquid syringe, alternately inject three
20-40 ul portions each of standard and sample solutions. Measure
the peak height or peak area for each peak and calculate the
average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent butylate as follows:
7 - (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Eva Santos, EPA Region IX, San Francisco,
California.
-------
July 1975 Butylate EPA-2
(Tentative)
Determination of Butylate by
High Pressure Liquid Chromatography
Butylate is the common name for S-ethyl diisobutylthiocarbamate,
a registered herbicide having the chemical structure:
CH2— CH - CH3
Molecular formula: CiiH23ONS
Molecular weight: 217.4
Boiling point: 71°C at 10 mm
Physical state and color: Amber liquid
Solubility: 45 ppm in water at room temperature; miscible with kerosene,
acetone, methyl isobutyl ketone, ethanol, xylene
Stability: stable under ordinary conditions; non-corrosive
Other names: Sutan (Stauf fer) , R1910
Reagents;
1. Butylate standard of known % purity
2. Chloroform
3. Dichlorome thane
4 . Hexane
5 . Methanol
All solvents should be pesticide or spectro grade.
-------
2 Butylate EPA-2
(Tentative)
Equipment:
1. High Pressure Liquid Chromatograph
2. High pressure liquid syringe or sample injection loop
3. Liquid chromatographic column, 4 mm I.D. x 25 cm packed
with LiChrosorb Si 60 - 10 u (or equivalent column)
Operating conditions for Hewlett-Packard Model 1010B LC:
Mobile phase: 80% Hexane + 19% Dichloromethane + 1% Methanol
Column temperature: ambient
2
Observed column pressure: 30 Kg/cm (425 PSI)
Flow rate: 3 ml/min
Detector: UV at 240 nm
Chart speed: 0.5 in/min
Injection: 10 pi
Conditions may have to be varied by the analyst for other instru-
ments, column variations, sample composition, etc. to obtain optimum
response and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.04 gram butylate standard into a 50 ml volumetric
flask; dissolve and make to volume with chloroform (final cone
0.8 mg/ml).
Preparation of Sample:
Weigh an amount of sample equivalent to 0.08 gram butylate
into a 100 ml volumetric flask; dissolve and make to volume
with chloroform (final cone 0.8 mg butylate/ml).
-------
3 Butylate EPA-2
(Tentative)
De terminat ion:
Using a high pressure liquid syringe, alternately inject
three 10 ul portions each of standard and sample solutions.
Measure the peak height or peak area for each peak and calculate
the average for both standard and sample.
Adjustments in attenuation or amount injected may have to
be made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the
percent butylate as follows:
„ _ (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method developed by Joseph B. Audino, Supervisor, Pesticide Formulation
Laboratory, California Department of Food and Agriculture; and by
Yoshihiko Kawano, Associate Chemist on sabbatical leave from the
University of Hawaii.
-------
September 1975 Butylate EPA-3
(Tentative)
Determination of Butylate
by Gas-Liquid Chroraatography (FID)
Butylate is the common name for S-ethyl diisobutylthiocarbamate,
a registered herbicide having the chemical structure:
CH2 CH—CH3
CH3—CH2—S—C N
CH2 CH CH3
Molecular formula: C .H ONS
Molecular weight: 217.4
Boiling point: 71°C at 10 mm
Physical state and color: Amber liquid
Solubility: 45 ppm in water at room temperature; miscible with
kerosene, acetone, methyl isobutyl ketone, ethanol, xylene
Stability: stable under ordinary conditions; non-corrosive
Other names: Sutan (Stauffer), R1910
Reagents:
1. Butylate standard of known % purity
2. Acetone, pesticide or spectro grade
-------
2 Butylate EPA-3
(Tentative)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 1/4" glass column packed with 5% QF-1 on
60/80 Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 10 jil
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 140°
Injection temperature: 215°
Detector temperature: 225°
Carrier gas: Helium or Nitrogen
Flow rate: 55 ml/min
Operating conditions for column temperature, carrier gas flow,
or hydrogen/air flow rates should be adjusted by the analyst to
obtain optimum response and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.05 gram butylate standard into a 25 ml volumetric
flask; dissolve and make to volume with acetone. (final cone
2 mg butylate/ml)
-------
Butylate EPA-3
(Tentative)
Preparation of Sample;
For technical material and liquid formulations, weigh a
portion of sample equivalent to 0.05 gram butylate into a 25 ml
volumetric flask, make to volume with acetone,and mix thoroughly.
(final cone 2 mg butylate/ml)
For dry formulations, weigh a portion of sample equivalent
to 0.5 gram of butylate into a 125 ml screw-cap flask, add by
pipette 50 ml acetone, and shake for one hour. Allow to settle;
filter or centrifuge if necessary, taking precautions to prevent
evaporation. Pipette 5 ml of the clear solution into a 25 ml
volumetric flask and make to volume with acetone and mix
thoroughly. (final cone 2 mg butylate/ml)
Determination;
Using a precision liquid syringe, alternately inject three
2-4 ul portions each of standard and sample solutions. Measure
the peak height or peak area for each peak and calculate the
average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent butylate as follows:
_, _ (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Eva Santos, EPA Region IX, San Francisco,
California.
-------
October 1975
Butylate EPA-4
Determination of Butylate by
Gas-Liquid Chromatography
(FID - Internal Standard)
Butylate is the common name for S-ethyl diisobutylthiocarbamate,
a registered herbicide having the chemical structure:
CH3—
0
C
Molecular formula: C H ONS
CH2— CH — CH3
CH2 — CH - CH3
CH3
Molecular weight: 217.4
Boiling point: 71°C at 10 mm
Physical state and color: Amber liquid
Solubility: 45 ppm in water at room temperature; miscible with kerosene,
acetone, methyl isobutyl ketone, ethanol, xylene
Stability: stable under ordinary conditions; non-corrosive
Other names: Sutan (Stauf fer) , R1910
Reagents ;
1. Butylate standard of known % purity
2. S-Ethyl dipropylthiocarbamate (EPTC) standard of known 70 purity
3. Carbon disulfide, pesticide or spectro grade
4. Chloroform, pesticide or spectro grade
-------
2 Butylate EPA-4
Reagents (Cont.)'
5. Methanol, pesticide or spectro grade
6. Internal Standard solution - weigh 0.2 gram EPIC into a 50 ml
volumetric flask; dissolve in and make to volume with a
solvent mixture consisting of 80% carbon disulfide + 15%
chloroform -I- 5% methanol. (cone 4 mg EPIC/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 4 mm glass column packed with 3% OV-1 on
60/80 Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 130°C
Injection temperature: 225°C
Detector temperature: 250°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi
Hydrogen pressure: 20 psi
Air pressure: 30 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Butylate EPA-4
Procedure:
Preparation of Standard:
*
Weigh 0.08 gram butylate standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 20 ml of the internal
standard solution and shake to dissolve. (final cone 4 mg butylate
and 4 mg EPIC/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.08 gram butylate
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the butylate. For
coarse or granular materials, shake mechanically for 10-15
minutes or shake by hand intermittently for 25-30 minutes.
(final cone 4 mg butylate and 4 mg EPTC/ml)
Determination:
Inject 1-2 jil of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from
1/2 to 3/4 full scale. The elution order is EPIC, then butylate.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of butylate and EPTC from
both the standard-internal standard solution and the sample-
internal standard solution.
-------
4 Butylate EPA-4
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
RF m (wt. EPTC)(% purity EPTC)(pk. ht. or area butylate)
(wt. butylate) (% purity butylate) (pk. ht. or area EPTC)
Determine the percent butylate for each injection of the
sample-internal standard solution as follows and calculate the
average :
7 , (wt. EPTC)(% purity EPTC) (pk. ht. or area butylate) (
° " (wt. sample) (pk. ht. or area EPTC) (RF) • t4~l)
Method submitted by Division of Regulatory Services, Kentucky
Agricultural Experiment Station, University of Kentucky, Lexington,
Kentucky 40506.
-------
October 1975 Butylate EPA-5
(Tentative)
Determination of Butylate
by Gas— Liquid Chromatography
(TCD - Internal Standard)
Butylate is the common name for S-ethyl diisobutylthiocarbamate,
a registered herbicide having the chemical structure:
0 CH2— CH - CH3
CH3 — CH2 — S— C — N<^
CH2 — CH - CH3
Molecular formula: C H ONS
Molecular weight: 217.4
Boiling point: 71°C at 10 mm
Physical state and color: Amber liquid
Solubility: 45 ppm in water at room temperature; miscible with
kerosene, acetone, methyl isobutyl ketone, ethanol, xylene
Stability: stable under ordinary conditions; non-corrosive
Other names: Sutan (Stauf f er) , R1910
Reagents:
1. Butylate standard of known % purity
2. Vernolate standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.25 gram vernolate into a
25 ml volumetric flask, dissolve in, and make to volume with
acetone, (cone 10 mg vernolate/ml)
-------
2 Butylate EPA-5
(Tentative)
Equipment^
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 5' x 1/4" glass column packed with 5% PEG-1540
on 60/80 Chromosorb W AW DMCS
3. Precision liquid syringe: 25 or 50 pi
4. Usual laboratory glassware
Operating Conditions for TCD;
Column temperature: 150°C
Injection temperature: 200°C
Detector temperature: 200°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas flow rate: 30 ml/min
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response and
reproducibility.
Procedure;
Preparation of Standard:
Weigh 0.1 gram butylate standard into a small glass-stoppered
flask or screw-cap tube. Add by pipette 10 ml of the internal
standard solution and shake to dissolve, (final cone 10 mg
butylate and 10 mg vernolate/ml)
-------
Butylate EPA-5
(Tentative)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.1 gram butylate
into a small glass-stoppered flask or screw-cap tube. Add by
pipette 10 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the butylate. For
coarse or granular materials, shake mechanically for 30 minutes
or shake by hand intermittently for one hour, (final cone 10 mg
butylate and 10 mg vernolate/ml)
Determination;
Inject 10-20 ^il of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from
1/2 to 3/4 full scale. The elution order is butylate, then
vernolate.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of butylate and vernolate
from both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. vernolate)(% purity vernolate)(pk. ht. or area butylate)
(wt. butylate)(% purity butylate)(pk. ht. or area vernolate)
Determine the percent butylate for each injection of the
sample-internal standard solution as follows and calculate the average:
„ _ (wt. vernolate) (% purity vernolate) (pk. ht. or area butylate) (iW)
(wt. sample)(pk. ht. or area vernolate)(RF) t I"'
-------
Butylatc EPA-5
(Tentative)
This method was submitted by the Commonwealth of Virginia, Division
•
of Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
Note! This method has been designated as tentative since it is a
Va. Exp. method and because some of the data has been suggested
by EPA's Beltsville Chemistry Lab. Any comments, criticism,
suggestion, data, etc. concerning this method will be appreciated.
-------
January 1976 Cadmium Compounds EPA-1
Determination of Cadmium in Fungicide Formulations
by Atomic Absorption Spectroscopy
Cadmium compounds such as the carbonate, chloride, oxide, sebacate,
succinate, and sulfate are registered turf fungicides.
Cadmium is a silver-white, blue-tinged, lustrous metal: atomic
symbol, Cd; atomic weight, 112.40; m.p. 321°C, b.p. 767°C, and d. 8.65.
It is insoluble in water; readily soluble in dilute HNO,; slowly solu-
ble in hot HC1; almost unattacked by cold, but converted into the
sulfate by hot H SO . It is present to the extent of 49 to 87% in the
above compounds.
Principle and Applicability of the Method;
This method is applicable for the analysis of cadmium in the
presence of organic materials and in combination with dithiocarbamates,
potassium chromate, coloring materials, and diluents. Using only a
simple acid digestion and filtration with no need for any special
extraction or clean-up procedures, most samples require less than 1 hour
from weighing to analysis.
The secondary absorption at 326.1 run is used for macro amounts of
cadmium in formulations rather than the most sensitive absorption at
228.0 run which is normally used for micro amounts.
Reagents;
1. Cadmium carbonate of known % cadmium
2. Concentrated nitric acid, ACS
3. Distilled or de-ionized water, free from metals
-------
2 Cadmium Compounds EPA-1
Equipment I
1. Atomic absorption spectrophotometer
2. Hot plate
3. Filtration apparatus
4. Whatman No. 42 (or equivalent) filter paper
5. Usual laboratory glassware
Procedure:
Preparation of Standard Solutions;
Standard solutions in the range of 100-500 ppm cadmium can be
made from separate weighings of cadmium carbonate (0.1534 gram
for each 100 ppm cadmium when made to 1 liter volume); however,
it is more convenient to prepare a 1000 ppm stock solution and
make appropriate dilutions.
A stock solution of 1000 ppm cadmium is made as follows:
weigh 1.534 grams of cadmium carbonate into a 150 ml beaker, add
15 ml concentrated nitric acid, and cover with a watch glass.
Boil gently to expel excess acid, cool, transfer to a 1000 ml
volumetric flask and make to volume with water. Prepare solutions
of 100, 200, 300, 400, and 500 ppm by diluting 10, 20, 30, 40, and
50 ml aliquots to 100 ml.
Preparation of Sample;
Weigh a portion of sample equivalent to 0.02-0.03 gram cadmium
into a 150 ml beaker, add 15 ml concentrated nitric acid, and cover
with a watch glass. After the initial reaction subsides and the
vapor above the solution is pale yellow, carefully add 15 ml water
and heat on a hot plate until the volume is reduced to approximately
15-20 ml. While the solution is still hot, filter through a
Whatman No. 42 filter paper and wash with 50-60 ml water (equivalent
-------
3 Cadmium Compounds EPA-1
filter papers may be used if they retain the fine materials normally
associated with clay carriers). Cool the filtrate; transfer to a
100 ml volumetric flask and make to volume with water.
Determination;
Following the manufacturer's manual for cadmium determination
for the particular instrument being used, proceed as follows:
Allow atomic absorption spectrophotometer to warm up one-half
hour and adjust Boling burner head so that the top of the oxidizing
flame lies approximately 1" below the center of the hollow cathode
tube. Regulate flame (acetylene and air are approximately 9 psig);
determine the precise maximum for secondary absorption, using the
most dilute standard, while holding lamp current at 6 ma. The 6 ma
value is chosen to minimize any auto-absorption which has been
reported in cadmium analysis. Aspirate standards and plot their
absorbances against concentration in ppm.
Determine concentrations of samples from plotted values of
standards. On some instruments where ppm may be read directly from
a digital readout, the plotting of an absorbance-concentration curve
is not necessary.
Using the above procedure, Beer's law is obeved in the 100-500
ppm range.
Calculation:
- raAn*m = (PP* Cd)(10~6)(100)
7« Cadmium = _, ,. > ,, ,,,„,.
tgrains sample) (l/iuu;
Method developed by Paul D. Jung and David Clarke, Division of Inspection
and Regulation, Maryland Department of Agriculture, College Park, Md.
20742 (method published JAOAC Vol. 57, No. 2, 1974, pg. 379-381).
-------
September 1975 Captafol EPA-1
(Tentative)
Determination of Captafol
by Infrared Spectroscopy
Captafol is the common name for cis-N-((1,1,2,2-tetrachloroethyl-
thio)-4-cyclohexene-l,2-dicarboximide, a registered fungicide having
the chemical structure:
0
HC" XH—C Cl Cl
\ I I
N—S—C—C-H
HC^ ^~\ Cl Cl
Molecular formula: C -H Cl NO.S
Molecular weight: 349.1
Melting point: 160 to 161°C
Physical state and color: white crystalline solid; technical material
is a light tan powder with a characteristic
odor.
Solubility: practically insoluble in water, slightly soluble in most
organic solvents
Stability: stable except under strongly alkaline conditions, slowly
decomposes at its melting point
Other names: Difolatan (Chevron Chem. Co.), Folcid
Reagents;
1. Captafol standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
Captafol EPA-1
(Tentative)
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm KBr or NaCl cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure i
Preparation of Standard;
Weigh 0.08 gram captafol standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml chloroform by
pipette, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 8 rag/ml)
Preparation of Sample;
For granular or dust formulations, weigh a portion of sample
equivalent to 0.4 gram captafol into a glass-stoppered flask or
screw-cap bottle. Add 50 ml chloroform by pipette and 1-2 grams
anhydrous sodium sulfate. Close tightly and shake for one hour.
Allow to settle; centrifuge or filter if necessary, taking pre-
caution to prevent evaporation. (final cone 8 mg captafol/ml)
For flowable liquid (water) formulations, weigh a portion of
sample equivalent to 0.4 gram captafol into a glass-stoppered flask
or screw-cap bottle. Add 50 ml chloroform by pipette, a few boiling
chips to aid agitation, and sufficient anhydrous sodium sulfate to
absorb all the water. Close tightly and shake vigorously on a
shaking machine for one hour. Allow to settle. Filter or centri-
fuge if necessary to get a clear chloroform solution, taking pre-
caution to prevent evaporation, (final cone 8 mg captafol/ml)
-------
Captafol EPA-1
(Tentative)
Determination:
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular 1R instrument
being used, scan both the standard and sample from 2000 cm~ to
1540 cm"1 (5.0 u-6.5 ji).
Determine the absorbance of standard and sample using the
peak at 17.27 cm" (5.79 p) and baseline from 18.18 cm to
1639 cm"1 (5.5 p to 6.1 ft).
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent captafol as follows:
2 m (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method submitted by Eva Santos, EPA Product Analysis Laboratory,
Region IX, San Francisco, California.
-------
September 1975
Captan EPA-1
Determination of Captan by
the Hydrolyzable Chlorine Method
Captan is the common name for N-trichloromethylthio-4-cyclo-
hexene-l,2-dicarboximide, a registered fungicide having the chemical
structure:
HC
HC
'CH-
0
.CH-
i—s—c—a
I
Cl
H2
Molecular formula: C-H-C1.NO-S
7 O J f.
Molecular weight:
Melting point:
300.6
178°C (decomposes)
Physical state and color: white crystalline solid; technical material
is a yellow amorphous solid (with a pungent odor)
of 93-95% purity and m.p. 160-170°C
Solubility: less than 0.5 ppm in water at RT; insoluble in petroleum
oils; at 25°C the solubility w/w is 7% in xylene, 5% in
chloroform, 3% in acetone, 1% in isopropanol
Stability: stable except under alkaline conditions; decomposes at
its melting point; non-corrosive but decomposition
products are corrosive
Other names: Orthocide (Chevron Chem. Co.), Merpan, Vondcaptan
-------
2 Captan EPA-1
This method is based on measuring the hydrolyzable chlorine in
captan and is designed for 100% captan. It has been used successfully
for high concentration captan formulations when there are no inter-
fering substances present. Any material containing hydrolyzable
chlorine would interfere. The chloride is measured on the sample
before and after hydrolysis and the difference calculated to equivalent
captan.
Reagents;
1. Absolute methanol
2. Acetone
3. Hydrogen peroxide, 30%
4. Nitric acid, 1+1
5. Sodium hydroxide, 0.25N
Equipment:
1. Potentiometric titrimeter
2. Reflux apparatus
3. Usual laboratory glassware
Procedure;
Preparation of Sample;
Weigh a portion of sample equivalent to 0.25 gram captan into
a 250 ml Erlenmeyer flask. Add 125 ml absolute methanol and swirl.
(Do not allow to stand more than 45 minutes before proceeding since
captan may slowly react with methanol.) Add acetone to the 250 ml
mark and mix thoroughly to dissolve the captan. Adjust the volume
as necessary because of solvent shrinkage, temperature change, etc.
(With technical captan and formulations a flocculent precipitate or
undissolved residue may be present.)
-------
3 Captan EPA-1
Transfer one 100 ml aliquot to a 500 ml standard taper
Erlenmeyer (or other suitable) flask to be refluxed for the
hydrolyzed chlorine content and another 100 ml aliquot to a
400 ml beaker for immediate titration of the non-hydrolyzed
chlorine content.
Hydrolysis and Determination of Hydrolyzed Chlorine;
Add 50 ml of approx. 0.25N sodium hydroxide solution and a
few boiling chips to the Erlenmeyer flask, connect to an upright
condenser, and reflux for one hour.
(Titrate the non-hydrolyzed aliquot at this time.)
Turn off or remove the heat and cautiously add 5 ml 30Z
hydrogen peroxide thru the condenser. Cool somewhat and remove
the flask from the condenser. Boil for 10 minutes to decompose
the excess hydrogen peroxide and evaporate to about 60 ml. (If
the solution is not practically colorless, add 5 ml more hydrogen
peroxide and water if necessary to maintain the volume and boil
another 10 minutes.)
Cool, add 10 ml 1 + 1 nitric acid, and titrate the chlorine
potentiometrically.
Determination of Chlorine before Hydrolysis:
Add 10 ml 1 + 1 nitric acid to the 100 ml aliquot in the 400 ml
beaker and titrate the chlorine potentiometrically.
Calculation:
The percent captan is obtained by subtracting the milliequiv-
alents of chlorine found before hydrolysis (meq. Cln) from the
B
milliequivalents of total chlorine found after hydrolysis (meq. Cl_),
multiplying by the milliequivalent weight of captan X 100, and
dividing by the weight of sample X the aliquoting factor.
-------
Captan EPA-1
captan - (meq' C1T " meq* C1B
(weight sample)(0.4)
where: meq. C1T - N AgNO. X ml used for titration of
hydrolyzed aliquot
meq. Cln - N AgNO, X ml used for titration of
B J
non-hydrolyzed aliquot
meq. wt. captan - (3)(iSo) " °'1002
aliquoting factor - r - 0.4
-------
August 1975 Captan EPA-2
Determination of Captan
by Infrared Spectroscopy
Captan is the common name for.N-trichloromethylthio-4-cyclo-
hexene-l,2-dicarboximide, a registered fungicide having the
chemical structure:
Cl
— Cl
Cl
Molecular formula: C0H0C1,N00S
y o j 2.
Molecular weight: 300.6
Melting point: 178°C (decomposes)
Physical state and color: white crystalline solid; technical material
is a yellow amorphous solid(with a pungent odor)
of 93-95% purity and m.p. 160-170°C
Solubility: less than 0.5 ppm in water at RT; insoluble in petroleum
oils; at 25°C the solubility w/w is 7% in xylene, 5% in
chloroform, 3% in acetone, 10% in isopropanol
Stability: stable except under alkaline conditions; decomposes at
its melting point; non-corrosive but decomposition
products are corrosive
Samples containing malathion and methoxychlor should be run by GLC.
Reagents:
1. Captan standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
2 Captan EPA-2
Equipment;
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.1 ram NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, put in 1-2 grams anhydrous sodium sulfate, and
seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.1 gram captan standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml chloroform by pipette, and
shake to dissolve. Add a small amount of anhydrous sodium sulfate
to insure dryness. (final cone 10 mg/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.5 gram captan into a
glass-stoppered flask or screw-cap bottle. Add 50 ml chloroform
by pipette and 1-2 grams anhydrous sulfate. Close tightly and
shake for one hour. Allow to settle; centrifuge or filter if
necessary, taking precaution to prevent evaporation, (final cone
10 mg captan/ml) For very low percent formulations requiring
larger samples, use more solvent and evaporate an aliquot to a
smaller volume to give a final concentration close to 10 mg captan/ml,
-------
3 Captan EPA-2
De terminat ion;
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1885 cm to
1665 cm"1 (5.3 p to 6.0yu).
Determine the absorbance of standard and sample using the
peak at 1735 cm" (5.76 ^) and basepoint at 1855 cm" (5.39yu).
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent captan as follows:
-, _ (abs. sample) (cone, std in mg/ml) (_%_ jpurity std)
(abs. std)(cone, sample in mg/ral)
(A concentration of 1 mg captan/ral chloroform gives an absorbance
of approx. 0.04 in a 0.1 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Laboratory Services, 1 North 14th Street, Richmond, Virginia 23219.
-------
October 1975
Carbaryl EPA-1
Determination of Carbaryl
by Ultraviolet Spectroscopy
Carbaryl is the accepted common name for 1-naphthyl methylcar-
bamate, a registered insecticide having the chemical structure:
—C N — CH3
Molecular formula: C H NO
Molecular weight: 201.2
Melting point: 1428C
Physical state and color: white, crystalline solid
Solubility: 40 ppm in water at 30°C; soluble in most polar organic
solvents such as acetone, dimethylformamide
Stability: stable to light, heat, and hydrolysis under normal storage
conditions; non-corrosive to metals, packaging materials,
or application equipment; compatible with most pesticides
except those strongly alkaline which hydrolyze it to
1-naphthol
Other names: Sevin (Union Carbide), sevin (USSR), UC 7744, Hexavin
Karbaspray, Ranyon, Septene, Tricarnam
This method is recommended only when the preferred infrared method
(AOAC 12th Ed., 2nd Supplement, 6.B01-6.B04) cannot be used.
-------
2 Carbaryl EPA-1
Reagents;
1. Carbaryl standard of known % purity
2. Ethanol, pesticide or spectro grade
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Filtration apparatus
4. Usual laboratory glassware
Procedure;
Preparation of Standard;
Weigh 0.05 gram carbaryl standard into a 100 ml volumetric
flask. Dissolve, make to volume with ethanol, and mix thor-
oughly. Pipette a 10 ml aliquot into a 50 ml volumetric flask
and make to volume with ethanol. Mix thoroughly and pipette a
10 ml aliquot into a 50 ml volumetric flask. Make to volume
and again mix thoroughly. (final cone 20 jig/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.05 gram of carbaryl
into a 250 ml Erlenmeyer glass-stoppered flask. Add 100 ml ethanol
by pipette and shake on a mechanical shaker for one hour. Filter
if necessary and pipette 10 ml of the clear filtrate into a 50 ml
volumetric flask. Make to volume with ethanol, mix thoroughly,
and pipette 10 ml into a 50 ml volumetric flask. Make to volume
with ethanol and mix thoroughly, (final cone 20 ^ig carbaryl/ml)
-------
3 Carbaryl EPA-1
UV Determination:
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 280 nm with
ethanol in each cell. Scan both the standard and sample from
350 nm to 250 nm with ethanol in the reference cell. Measure
the absorbance of both standard and sample at 280 nm.
Calculation;
From the above absorbances and using the standard and
sample concentrations, calculate the percent carbaryl as follows:
„ m (abs. sample)(cone. std in ug/ml)(% purity std)
(abs. std) (cone, sample in jig/ml)
-------
October 1975
Carbaryl EPA-2
(Tentative)
Determination of Carbaryl by
High Pressure Liquid Chromatography
Carbaryl is the accepted common name for 1-naphthyl tnethylcar-
bamate, a registered insecticide having the chemical structure:
0 H
Molecular formula: Ci2HllN02
Molecular weight: 201.2
Melting point: 142°C
Physical state and color: white, crystalline solid
Solubility: 40 ppm in water at 30°C; soluble in most polar organic
solvents such as acetone, dimethylformamide
Stability: stable to light, heat, and hydrolysis under normal storage
conditions; non-corrosive to metals, packaging materials,
or application equipment; compatible with most pesticides
except those strongly alkaline which hydrolyze it to
1-naphthol
Other names: Sevin (Union Carbide), sevin (USSR), UC 7744, Hexavin
Karbaspray, Ranyon, Septene, Tricarnam
-------
2 Carbaryl EPA-2
(Tentative)
Reagents;
1. Carbaryl standard of known % purity
2. Methanol, pesticide or spectro grade
Equipment;
1. High pressure liquid chromatograph with UV detector at
254 nm. If a variable wavelength UV detector is available,
other wavelengths may be useful to increase sensitivity or
eliminate interference.
2. Suitable column such as:
a. DuPont ODS Perraaphase, 1 meter x 2.1 mm ID
b. Perkin Elmer Sil-X 11 RP, 1/2 meter x 2.6 mm ID
3. High pressure liquid syringe or sample injection loop
4. Usual laboratory glassware
Operating Conditions;
Mobile phase: 20% methanol + 80% water
Column temperature: 50-55°C
Chart speed: 5 min/inch or equivalent
Flow rate: 0.5 to 1.5 ml/min (Perkin Elmer instrument
with 1/2 meter column)
Pressure: 400 psi (DuPont instrument with 1 meter column)
Attenuation: adjusted
Conditions may have to be varied by the analyst for other
instruments, column variations, sample composition, etc. to obtain
optimum response and reproducibility.
-------
Carbaryl EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.3 gram carbaryl standard into a glass-stoppered
flask or screw-cap bottle, add 100 ml methanol by pipette,
dissolve, and mix well (final cone 3 mg/ral).
Preparation of Sample;
Weigh an amount of sample equivalent to 0.3 gram carbaryl
into a glass-stoppered flask or screw-cap bottle, add 100 ml
methanol by pipette,and shake thoroughly to dissolve the
carbaryl. Allow any solid matter to settle; filter or centri-
fuge if necessary (final cone 3 mg carbaryl/ml).
Determination:
Alternately inject three 5 ul portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent carbaryl as follows:
7 - (pk. ht. or area sample)(wt. std injected)(% purity std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA, Beltsville, Md.
-------
September 1975
Carbofuran EPA-1
Determination of Carbofuran
by Infrared Spectroscopy
Carbofuran is the accepted common name for 2,3-dihydro-2,2-
dimethyl-7-benzofuranyl methyl carbamate, a registered insecticide,
acaricide, and nematocide having the chemical structure:
CH2
Molecular formula: ci2HisN03
Molecular weight: 221.3
Melting point: 150-152°C
Physical state, color, and odor: Odorless, white, crystalline solid
Solubility: solubility at 25°C is 700 ppm in water, 15% in acetone,
14% in acetonitrile, 4% in benzene, 9% in cyclohexanone,
27% in dimethylformamide
Stability: stable under neutral or acid conditions, unstable in
alkaline media
Other names: Furadan (Niagara), NIA 10242, Bay 70142, FMC 10242,
Curaterr
Reagents;
1. Carbofuran standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
2 Carbofuran EPA-1
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.2 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, put in 1-2 grams anhydrous sodium sulfate, and
seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.15 gram carbofuran standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml chloroform by
pipette, close tightly, and shake to dissolve. Add a small
amount of anhydrous sodium sulfate to insure dryness. (final
cone 15 mg/ml)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.75 gram carbofuran
into a glass-stoppered flask or screw-cap tube. Add 50 ml
chloroform by pipette and 1-2 grams anhydrous sodium sulfate.
Close tightly and shake for one hour. Allow to settle; centrifuge
or filter if necessary, taking precautions to prevent evaporation.
(final cone 15 mg carbofuran/ml)
-------
3 Carbofuran EPA-1
Determination ;
With chloroform In the reference cell, and using the optimum
quantitative analytical settings, scan both the standard and
le from 1000 cm" to 800 cm~ (10 ji to 12.5 ji) .
Determine the absorbance of standard and sample using the
at 875 cm
(11.11 u to 11.83
sample from 1000 cm" to 800 cm~ (10 ji to 12.5
the absorbance of standard and sa
peak at 875 cm (11.93 ) and baseline from 900 cm"1 to 845 cm
(The N-H band at 3460 cm" (2.89 ji) is also very good.)
Calculation;
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent carbofuran as
follows:
„ m (abs. sample) (cone, std in mg/ml) (% purity std)
(abs. std) (cone, sample in mg/ml)
(A concentration of 1 mg carbofuran/ml chloroform gives an
absorbance of approx. 0.02 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
-------
November 1975
Carboxin EPA-1
(Tentative)
Determination of Carboxin in Dusts
and Powders by Infrared Spectroscopy
Carboxin is the common name for 5,6-dihydro-2-methyl-l,4-oxathiin-
3-carboxanilide, a registered fungicide having the chemical structure:
Molecular formula: C H NO S
Molecular weight: 235
Melting point:
91.5 to 92.5°C; a dimorphic form has a m.p. of 98 to 100°C
Physical state, color, and odor: odorless, white, crystalline solid
(The technical product is at least 97% pure.)
Solubility: 170 ppm in water at 25°C; soluble in acetone, benzene,
dimethyl sulfoxide, ethanol, methanol
Stability: compatible with all except highly alkaline or acidic pesticides
Other names: Vitavax, D735 (Uniroyal); DCMO
Reagents:
1. Carboxin standard of known % purity
2. Benzene, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
Carboxin EPA-1
(Tentative)
Equipment:
1. Infrared spectrophotometer, double beam ratio recording,
with matched 0.2 mm NaCl or KBr cells
2. Mechanical shaker
3. Soxhlet extraction apparatus
4. Rotary evaporator or steam bath with short reflux column
5. Filtration apparatus or centrifuge
6. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.08 gram carboxin standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml benzene by pipette, and shake
to dissolve. Add a small amount of anhydrous sodium sulfate to
insure dryness. (final cone 8 mg/ml)
Preparation of Sample;
For Shake-out extraction, weigh a portion of sample equivalent
to 0.8 gram carboxin into a 250 ml glass-stoppered or screw-cap
Erlenmeyer flask. Add by pipette 100 ml benzene, stopper tightly,
and shake on a mechanical shaker for 2 hours. Allow to settle?
filter or centrifuge if necessary, taking precaution to prevent
evaporation. (final cone 8 mg carboxin/ml)
For Soxhlet extraction, weigh a portion of sample equivalent to
0.8 gram carboxin into a Soxhlet thimble, plug with cotton or glass
wool, and extract with benzene for 3 hours. Evaporate to a suitable
volume under vacuum on a rotary evaporator or on a steam bath using
a short reflux column. Quantitatively transfer to a 100 ml volu-
metric flask and make to volume with benzene. Add a small amount of
granular anhydrous sodium sulfate to insure dryness. (final cone
8 mg carboxin/ml)
-------
Carboxin EPA-1
(Tentative)
Determination:
With benzene in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1820 cm to
1110 cm'1 (5.5 ji to 9.0 ji) .
Determine the absorbance of standard and sample at either of
the following three bands:
Peak Basepoint
1675 cm"1 (5.97 p) 1630 cm"1 (6.13 ji)
1585 cm"1 (6.30p) 1630 cm"1 (6.13^i)
1290 cm"1 (7.75 ;i) 1265 cm"1 (7.90;i)
Either of these bands may be used with comparable results.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent carboxin as follows:
7 = (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
This method is based on an EPA experimental method using data from
Uniroyal. Any suggestions, data, criticisms, and information on its
use will be appreciated.
-------
October 1975
Chlorbromuron EPA-1
(Tentative)
Determination of Chlorbromuron by
Gas-Liquid Chromatography (FID)
Chlorbromuron is the accepted, common name for 3-(4-bromo-3-chloro-
phenyl)-l-methoxy-l-methylurea, a registered herbicide having the
chemical structure:
0—CH3
Molecular formula: C H BrClN-0
Molecular weight: 293.6
Melting point: 97°C (The technical grade has a purity of 95%
and melts at 90-95°C)
Physical state, color, and odor: off-white crystalline solid with a
mild odor
Solubility: 50 ppm in water at RT; soluble in acetone, chloroform,
methyl ethyl ketone, dimethylformamide; slightly soluble
in xylene
Stability: stable at RT; non-corrosive; compatible with other WP
formulations
Other names: Bromex (Nor-Am), Maloran (CIBA-GEIGY), chlorobromuron
(France)
-------
2 Chlorbromuron EPA-1
(Tentative)
Reagents:
1. Chlorbromuron standard of known % purity
2. Acetone, pesticide or spectro grade
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 2' x 1/4" glass column packed with 2% SE-52 on
70/80 Anakrotn ABS (or equivalent column)
3. Precision liquid syringe: 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 180°C
Injection temperature: 225°C
Detector temperature: 225°C
Carrier gas: Helium or Nitrogen
Flow rate: 55 ml/min
Operating parameters (above) as well as hydrogen/air flow rates,
attenuation, and chart speed should be adjusted by the analyst to
obtain optimum response and reproducibility.
Procedure:
Preparation of Standard;
Weigh 0.1 gram Chlorbromuron standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml acetone by pipette,
close tightly and shake to dissolve. (final cone 10 mg/ml)
-------
Chlorbromuron EPA-1
(Tentative)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.5 gram chlorbromuron
into a glass-stoppered flask or screw-cap bottle, add 50 ml
acetone by pipette, close tightly, and shake for one hour. Allow
to settle; filter or centrifuge if necessary,taking precautions
to prevent evaporation. (final cone 10 mg chlorbromuron/ml)
Determination:
Using a precision liquid syringe, alternately inject three
2-3 ul portions each of standard and sample solutions. Measure
the peak height or peak area for each peak and calculate the
average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent chlorbromuron as follows:
y - (pk. ht. or area sample)(wt. std injected)(% purity std)
(pk. ht. or area std)(wt. sample injected)
This method is based on one of EPA's Experimental Methods (No. 10)
which was adapted from another method from Ciba.
Comments, suggestions, data, results, etc. on this method are most
welcome.
-------
March 1976 Chlorophenoxy Herbicides EPA-1
Definition, Structure, and Technical Data
The chlorophenoxy herbicides are a group of chemical compounds
consisting mainly of mono-, di-, or tri- chlorinated phenoxy acetic,
propionic, or butyric acids. These compounds are registered herbicides;
however, some uses are restricted.
Formulations of these compounds may contain alkali metal salts
which are marketed in the solid state or as concentrated aqueous solu-
tions containing 10 to 40% active ingredient calculated as the acid.
Salts with amines are almost exclusively aqueous solutions containing
40 to 70% active ingredient. The esters are marketed in the form of
emulsifiable concentrates and as oil solutions for aerial spraying.
Formulations may contain various substances such as wetting agents,
emulsifiers, anti-precipitation agents, etc.
Structure and technical data for 11 of these compounds is given
below. For each compound the common name is followed by the chemical
name, structure, physical and chemical data, and other names.
2,4-U (ISO, BSI, WSSA), 2,4-dichlorophenoxyacetic acid
Cl
0—CH2-COOH
Molecular formula: CQH,C1_0
O D 2
Molecular weight: 221.0
-------
Chlorophenoxy Herbicides EPA-1
Physical state, color and odor: white crystalline solid, odorless when
pure, otherwise slight phenolic odor
Melting point: 140 to 141°C (pure), 135 to 138°C (technical)
Solubility: about 600 ppm in water at 25°C; soluble in aqueous
alkali and in alcohols, ether, acetone; insoluble in
petroleum oils
Stability:
Other names:
non-hygroscopic but corrosive; forms salts and esters
of varying properties and stabilities
Agrotect, Amoxone, Aqua-Kleen, Chipco Turf Herbicide D,
Chloroxone, Crop Rider, Decamine, Bed-Weed, Dormone,
Esteron, Estone, Fernesta, Fernimine, Fernoxone, Ferxone,
Hedonal, Pennamine D, Salvo, Tributon, Vergemaster,
Vertron 2D, Visko-Rhop, Weedar, Weedone
Dichlorprop (ISO, BSI, WSSA), 2-(2,4-dichlorophenoxy) propionic acid
Cl
-CH—COOH
Molecular formula: CnH0Cl_0_
y o / j
Molecular weight: 235.1
Physical state, color, and odor: white crystalline solid, odorless
when pure; technical, slight phenolic odor and tan
color
Melting point:
117.5 to 118.1°C (pure), 114 to 117°C (technical)
Solubility: about 350 ppra in water at 20°C; soluble in most organic
solvents
-------
Chlorophenoxy Herbicides EPA-1
Stability:
Other names:
acid is stable to heat and resistant to reduction, hydrolysis,
and atmospheric oxidation
Cornox RK (Boots Co. ltd), RD 406, 2,4-DP (USSR), Weedone
2,4-DP, Weedone 170, Envert 171
2,4-DB (BSI, WSSA), 4-(2,4-dichlorophenoxy) butyric acid
0-CH2-CH2-CH2-COOH
Molecular formula: C..QH.. Cl-O-
Molecular weight: 249.1
Physical state, color, and odor: white crystalline solid, odorless when
pure
Melting point: 117 to 121°C depending on purity
Solubility: practically insoluble in water; slightly soluble in
benzene, toluene, and kerosene; very soluble in acetone,
alcohol, and ether
Stability: acids, salts, and esters are stable
Other names: Embutox (May & Baker Ltd), Butoxone (Chipman), Butyrac
(Amchem), MB 2878
-------
Chlorophenoxy Herbicides EPA-1
2,4,5-T (ISO, BSI, WSSA), 2,4,5-trichlorophenoxy acetic acid
CI
0 —CH2—COOH
Molecular formula: C H Cl 0
O J J J
Molecular weight: 255.5
Physical state and color: white crystals
Melting point: 156.6°C (pure), 150-151°C (technical)
Solubility:
about 278 ppm in water at 25°C; soluble in acetone,
ethanol, and ether; salts with alkali metals and
amines are water-soluble but oil-insoluble; esters
are oil-soluble but water-insoluble
stable and non-corrosive
Weedone 2,4,5-T (Arachem), Brush-Rhop (Transvaal Inc.),
Estron 245 (Dow), Decamine, Ded-Weed Brush Killer,
Fence Rider, Forron, Fruitone A, Inverton 245, Line
Rider, Reddon, Tormona, Tributon, Trioxone, Weedar
Silvex (WSSA, ANSI), 2-(2,4,5-trichlorophenoxy) propionic acid
Stability:
Other names:
0—CH—COOH
CH3
-------
Chlorophenoxy Herbicides EPA-1
Molecular formula: CgH Cl 0
Molecular weight: 269.5
Physical state, color, and odor: white powder, low odor
Melting point: 179 to 181°C
Solubility:
Stability:
about 140 ppm in water at 25°C; soluble in acetone
and methanol
non-corrosive to spray equipment
Other names: ferroprop (common name ISO and BSI) , Kuron (Dow),
Weedone 2,4,5-TP (Amchem), Aqua-Vex, Ded-Weed,
Fruitone T, GarIon, Kurosal, 2,4,5-TP
2,4,5-TB (ISO), 4-(2,4,5-trichlorophenoxy) butyric acid
Cl
0-CH2-CH2-CH2-COOH
Molecular formula: C-0H_C1,0,
Molecular weight: 283.5
Physical state and color: white crystals
Melting point: 114 to 115°C
Solubility: similar to other compounds of this group
Stability: similar to other compounds of this group
Other names: 4-2,4,5-TB
-------
Chlorophenoxy Herbicides EPA-1
MCPA (BSI, WSSA),
(2-methyl-4-chlorophenoxy) acetic acid
~CH2—COOH
Molecular formula: C H CIO
Molecular weight: 200.6
Physical state and color: white crystalline solid (pure), light
brown solid (technical)
Melting point: 118 to 119°C (pure), 100 to 115°C (technical)
Solubility: about 825 ppm in water at RT; soluble in ethanol and
ether; forms water-soluble salts with alkali metals and
organic bases; oil-soluble esters may be prepared
Stability: solutions of alkali metals are alkaline and will corrode
aluminum and zinc; water-soluble salts may be precipitated
by hard water
Other names: Agroxone (Plant Protection Ltd); Agritox (May & Baker Ltd);
Cornox M (The Boots Co. Ltd); Chiptox, Rhomene, Rhonox
(Chipman Div. Rhodia Inc.); metaxon (USSR); Bordermaster;
Hormotuho; Kilsem; MCP; Mephanac; Zelan
Mecoprop (ISO, BSI, WSSA), 2-(2-methyl-4-chlorophenoxy) propionic acid
-------
Chlorophenoxy Herbicides EPA-1
Molecular formula: CioHllC103
Molecular weight: 214.6
Physical state, color, and odor: colorless, odorless, crystalline solid;
technical product may have a slight phenolic odor
Melting point: 94 to 95°C (technical 90°C or above)
Solubility:
about 620 ppm in water at 20°C; readily soluble in
most organic solvents; forms water-soluble salts
stable to heat; resistant to reduction, hydrolysis,
and atmospheric oxidation; corrosive to some metals
MCPP, CMPP, Iso-Comox (The Boots Co. Ltd), RD 4593,
Chipco Turf Herbicide MCPP, Hedonal MCPP, Kilprop,
Mepro, Methoxone
MCPB (WSSA), 4-(2-methyl-4-chlorophenoxy) butyric acid
Stability:
Other names:
0-CH2-CH2-CH2-COOH
Molecular formula: ciiHi3C103
Molecular weight: 228.5
Physical state and color: white solid
Melting point: 100 to 101°C (pure), 99 to 100°C (technical, about 90% purity)
Solubility: about 44 ppm in water at RT; slightly soluble in carbon
tetrachloride or benzene; soluble in acetone, alcohol,
and ether; forms water-soluble salts with alkali metals
-------
Chlorophenoxy Herbicides EPA-1
Stability:
Other names:
somewhat incompatible with hard water
Tropotox (May & Baker Ltd), MB 3046, Can-Trol (Chipman
Div. of Rhodia Inc.), Thistrol (Amchera), PDQ, 2,4-MCPB
(France), 2M-4Kh-M (USSR)
Erbon (ANSI, WSSA), 2-(2,4,5-trichlorophenoxy) ethyl-2,2-dichloropropionate
0 Cl
• 0-CH2-CH2-0-C—C-CH3
Cl
Molecular formula: C-jH Cl 0
Molecular weight: 366.5
Physical state and color: white solid (pure), dark brown solid (technical)
Melting point: 49 to 50°C; bp 161 to 164°C at 0.5 mm Hg
Solubility: practically insoluble in water; soluble in acetone,
ethanol, kerosene, xylene, and most oils
Stability: stable to UV light; non-flammable and non-corrosive
Other names: Baron, Erbon (Dow Chem. Co.)
-------
Chlorophenoxy Herbicides EPA-1
2,4-PEP (WSSA), a mixture of
tris [2-(2,4-dichlorophenoxy) ethyl] phosphite and
bis [2-(2,4-dichlorophenoxy) ethyl] phosphite
Cl
>-CH2-CH203P
0-CH2-CH202POH
tris form
bis form
Molecular formula: C0/H0.C1,0,P (tris), C.,H.1.C1/0,P (bis)
ZH 21 o o ID ij A 5
Molecular weight: 649.4 (tris), 460 (bis)
Physical state, color, and odor: dark amber viscous liquid with a
phenolic odor
Boiling point: above 200°C at 0.1 mm Hg
Solubility: practically insoluble in water; miscible with xylene
and aromatic hydrocarbons
Stability: stable when anhydrous; in presence of water or soil,
slowly hydrolyzed to 2,4-dichlorophenoxyethanol and
phosphoric acid; corrosive to iron and mild steel
Other names: Falone (Uniroyal), 3Y9
-------
March 1976 Chlorophenoxy Herbicides EPA-'
Determination of 2,4-D and 2,4,5-T in Formulations
by Ultraviolet Spectroscopy
For definition, structure, and technical data on these compounds,
see Chlorophenoxy Herbicides EPA-1. See note at end of method.
Principle of the Method:
A portion of sample is refluxed with sodium hydroxide whereby the
esters are saponified and the herbicide acids are converted into sodium
salts. The alkaline solution is extracted with ether to remove oils
and other organic solvent-soluble substances. The solution is then
acidified and the free herbicide acids are extracted with carbon tetra-
chloride which is evaporated. The herbicide acids are then dissolved
in sodium hydroxide solution and read in an ultraviolet spectrophotometer.
Reagents:
1. 2,4-D and/or 2,4,5-T standards of known % purity
2. Sodium hydroxide, 25% solution (freshly prepared)
3. Ethyl ether, ACS
4. Sulfuric acid, 1+1 solution
5. Carbon tetrachloride, ACS
6. Sodium hydroxide, IN solution
7. Sodium hydroxide, 0.1N solution (dilute above solution 1:10)
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Refluxing apparatus
-------
Chlorophenoxy Herbicides EPA-2
3. Filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.08 gram 2,4-D acid or 2,4,5-T acid (0.08 gram of each,
if both are present) into a 100 ml volumetric flask, dissolve in,
and make to volume with 0.1N sodium hydroxide solution. Mix
thoroughly and pipette 5 ml into a second 100 ml volumetric flask.
Make to volume with 0.1N sodium hydroxide solution and mix thor-
oughly, (final cone 40 ug 2,4-D and/or 2,4,5-T/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.04 gram 2,4-D acid
or 2,4,5-T acid (0.04 gram each, if both are present) into a
125 ml standard taper Erlenmeyer flask. Add 10 ml 25% sodium
hydroxide solution and several small glass beads. Attach to a
reflux condenser and reflux for at least one hour. Turn off the
heat, wash down the condenser with 10-15 ml water, remove from
apparatus, and cool to room temperature. Transfer the solution
quantitatively to a 125 ml separatory funnel, washing the Erlen-
meyer flask with 4-5 small portions of water.
Extract this solution with two 50 ml portions of ethyl ether.
Wash the ether extracts with two 10 ml portions of IN sodium
hydroxide solution and add the wash solutions to the alkaline
sample solution; discard the ether extracts. Neutralize the
alkaline sample solution carefully with 1+1 sulfuric acid and
add 1 ml in excess. The neutral point is indicated by precipitation
of the free organic acids.
Extract the acidified sample solution successively with 25, 15,
10, and 10 ml portions of carbon tetrachloride, shaking for 2-3
minutes each time. If the extracts are cloudy, combine in a 125 ml
-------
3 Chlorophenoxy Herbicides EPA-2
separatory funnel and clarify by washing with 10 ml water. Filter
the carbon tetrachloride extracts through a piece of cotton (wet
with carbon tetrachloride) into a 100 ml volumetric flask, make
to volume, and mix thoroughly.
Pipette 10 ml of the above solution into a 125 ml standard
taper Erlenmeyer flask and evaporate to dryness under vacuum,
warming in a water bath at about 40°C. Dissolve the residue in
10 ml of IN sodium hydroxide solution, transfer quantitatively to
a 100 ml volumetric flask,and make to volume with water. (final
cone 40 ug 2,4-D and/or 2,4,5-T/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 284 nm for 2,4-D
c3%>
or 2#£- nm for 2,4,5-T (296 nm when both are present) with 0.1N
sodium hydroxide solution in each cell. Scan both the standard
and sample from 350 nm to 250 nm with 0.1N sodium hydroxide
solution in the reference cell.
Measure the absorbance of standard and sample solutions at
284 nm for 2,4-D (secondary maximum 290 nm) and at 289 nm for
2,4,5-T (secondary maximum 296 nm).
Calculation:
Calculate the percent 2,4-D alone using the absorbance at
284 nm or the percent 2,4,5-T alone using the absorbance at 289 nm;
or, when both are present use the absorbance at 296 nm for 2,4,5-T.
7 = (abs. sample)(cone, standard injug/ml)(% purity standard)
(abs. standard)(cone, sample in ug/ml)
Note! Although this method is for 2,4-D and 2,4,5-T, it may be usable
for other chlorophenoxy herbicides. Data and comments on the use
of this method for other compounds, including linearity, accuracy,
and precision are most welcome by the Methods Editorial Committee.
-------
March 1976 Chlorophenoxy Herbicides EPA-3
(Tentative)
Determination of Chlorophenoxy Herbicide Acids and Esters
by High Pressure Liquid Chromatography
For definition, structure, and technical data on Chlorophenoxy
herbicide free acids, see Chlorophenoxy Herbicides EPA-1. (Data
and conversion factors for salts and esters of these compounds will
appear in supplements to this manual or in a later revised edition.)
Principle of the Method:
Formulations of Chlorophenoxy herbicides as esters or free acids
are dissolved in methanol and subjected to HPLC analysis using the
same column but different mobile phases. Esters are determined using
a 40% methanol-60% water mobile phase and the free acids are determined
using a 10% methanol-90% 0.0025M aqueous phosphoric acid mobile phase.
(Alkylamine salts have not been studied enough to include in this
method; however, a conversion into the free acid should allow HPLC
determination. Data and comments on analysis of these compounds would
be appreciated by the editorial committee.)
Reagents:
1. Chlorophenoxy herbicide acid or ester standards of known
% purity
2. Methanol - ACS
3. Phosphoric acid, 0.0025M aqueous solution
4. Ethyl ether - ACS
-------
2 Chlorophenoxy Herbicides EPA-3
(Tentative)
Equipment:
1. High pressure liquid chromatograph with UV detector at 254 nm.
If a variable wavelength UV detector is available, other
wavelengths may be useful to increase sensitivity or eliminate
interference. 235 nm has been found good for chlorophenoxy
herbicides.
2. Column: 1 meter x 2.1 mm ID stainless steel packed with
DuPont ODS Permaphase (or equivalent column such as
Perkin Elmer ODS Sil-X 11 RP)
3. High pressure liquid syringe or sample injection loop
4. Mechanical shaking apparatus
5. Usual laboratory glassware
Operating Conditions;
1. Mobile phase: esters - 40% methanol + 60% water
free acids - 10% methanol + 90% 0.0025M
aqueous phosphoric acid solution
2. Column temperature: 55°C
3. Pressure: 700-1000 psi (DuPont - constant pressure)
4. Flow rate: 0.5 to 1.5 ml/min (Perkin-Elmer - constant flow)
5. Chart speed: 5 minutes/inch or equivalent
6. Attenuation: adjust for 60-80% pen response for 5 ul injection
Conditions may have to be varied by the analyst for the specific
instrument being used, column variations, sample composition, etc. to
obtain optimum response and reproducibility.
-------
Chlorophenoxy Herbicides EPA-3
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.25 gram of chlorophenoxy herbicide acid or ester
standard into a 125 ml glass-stoppered flask or screw-cap bottle,
add 50 ml methanol by pipette, and shake to dissolve. (cone 5 mg/ml)
Preparation of Sample:
For liquid formulations, weigh a portion of sample equivalent
to 0.25 gram chlorophenoxy herbicide acid or ester into a 50 ml
volumetric flask; make to volume with methanol. (cone 5 mg/ml)
For solid formulations (powders or granules), weigh a portion
of sample equivalent to 0.5 gram chlorophenoxy herbicide acid or
ester into a 300 ml glass-stoppered flask or screw-cap bottle, add
200 ml ethyl ether, and shake on a mechanical shaker for one hour.
Allow to settle, pipette 20 ml into a small glass-stoppered flask,
and evaporate to a "moist" dryness. Add 10 ml methanol by pipette
and shake to dissolve residue. (final cone 5 mg/ml)
Determination:
Alternately inject three 5 ul portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent chlorophenoxy herbicide as follows:
7 - (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
-------
4 Chlorophenoxy Herbicides EPA-3
(Tentative)
Notes:
1. If a peak for a declared acid herbicide does not appear
using the 10% raethanol-90% 0.0025M phosphoric acid mobile
solvent, either it is not present or it is in the ester
form. This can be confirmed by changing the mobile phase
to 40% methanol-60% water to determine ester herbicides.
The reverse would be true if a declared ester herbicide
did not appear using the "ester mobile phase." A switch
to the "acid mobile phase" would then determine the acid
herbicide.
2. Due to the mixture of the branched heptyl radical with
methyl groups in the 3, 4, or 5 position, isooctyl esters
give peaks of varying patterns and cannot be analyzed. The
analysis will distinguish the isooctyl ester from the other
ester.
Method submitted by Elmer H. Hayes, EPA, Beltsville, Md,
-------
March 1976 Chlorophenoxy Herbicides EPA-4
(Tentative)
Determination of Butoxyethyl Esters of 2,4-D and 2,4,5-T
in Liquid Formulations by Gas-Liquid Chromatography (FID-IS)
For definition, structure, and technical data on 2,4-D and 2,4,5-T
acids, see Chlorophenoxy Herbicides EPA-1. (Data and conversion factors
for esters will appear in supplements or in a later revision of this
manual.)
Reagents;
1. 2,4-D butoxyethyl ester standard of known % purity
2. 2,4,5-T butoxyethyl ester standard of known % purity
3. Acetone, pesticide or spectro grade
4. Dibutyl phthalate, technical (or better)
5. Internal standard solution - weigh 0.2 gram dibutyl phthalate
into a 100 ml volumetric flask; dissolve in and make to
volume with acetone. (cone 2 mg dibutyl phthalate/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm ID glass, packed with 5% OV-210 on 80/100 mesh
Chromosorb W HP
3. Precision liquid syringe: 1 or 5 pi
4. Mechanical shaker or a Patterson-Kelley twin shell blender
that has been modified by replacing the blending shell with
a box to hold a 24-tube rubber-covered rack to hold 25 mm x
200 mm screw-top culture tubes
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
-------
Chlorophenoxy Herbicides EPA-4
(Tentative*
Operating Conditions for FID;
Column temperature: 175°C
•
Injection temperature: 225°C
Detector temperature: 225°C
Carrier gas: Nitrogen
Carrier gas flow rate: Adjusted for particular GC
Hydrogen flow rate: Adjusted for particular GC
Air flow rate: Adjusted for particular GC
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
Procedure;
Preparation of Standard:
Weigh 0.1 gram butoxyethyl ester of 2,4-D and/or 2,4,5-T
standard into a small glass-stoppered flask or screw-cap tube,
add 20 ml internal standard solution by pipette,and shake to
dissolve. (cone 5 mg 2,4-D and/or 2,4,5-T butoxyethyl esters
and 2 mg dibutyl phthalate/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.1 gram chlorophenoxy
herbicide (as above) into a glass-stoppered bottle or screw-cap
tube, add 20 ml internal standard solution by pipette, and shake
or tumble for one hour. Allow to settle; filter or centrifuge if
necessary. (cone 5 mg chlorophenoxy herbicide and 2 mg dibutyl
phthalate/ml)
-------
Chlorophenoxy Herbicides EPA-4
(Tentative)
Determination;
Inject 0.2-0.4 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from
1/2 to 3/4 full scale. The elution order is dibutyl phthalate,
2,4-D ester, and 2,4,5-T ester.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of the dibutyl phthalate
and the chlorophenoxy herbicides from both the standard-internal
standard solution and the sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
IS = Internal standard = dibutyl phthalate
CPH = Chlorophenoxy herbicide
„ (wt. IS)(% purity IS)(pk. ht. or area CPH)
= (wt. CPH)(% purity CPH)(pk. ht. or area IS)
Determine the percent CPH for each injection of the sample-
internal standard solution as follows and calculate the average:
% _ (wt. IS)(% purity IS)(pk. ht. or area CPH) (
(wt. sample) (pk. ht. or area IS)(RF)
This method was submitted by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond, Virginia 23219,
Note! This method has been designated as tentative since it is a Va. Exp.
method and because some of the data has been suggested by EPA's
Beltsville Chemistry Lab. Any comments, criticisms, suggestions,
data, etc. concerning this method and its use for other chloro-
phenoxy herbicides will be appreciated by the editorial committee.
-------
March 1976 Chlorophenoxy Herbicides EPA-5
(Tentative)
Determination of 2,4-D acid (1%) and Silvex acid (0.5%)
in Fertilizer Formulations by Gas-Liquid Chromatography
(FID-IS using on-column derivatization)
For definition, structure, and technical data on 2,4-D and
silvex, see Chlorophenoxy Herbicides EPA-1.
Principle of the Method;
The standard Chlorophenoxy herbicide and the extracted chloro-
phenoxy herbicide from the sample are made to a definite volume with
the internal standard solution, dibutyl phthalate in acetone. A
portion of either is injected along with a portion of the derivatizing
compound N-methyl-N-trimethylsilyl-2,2,2-trifluoroacetamide. The
formed derivative is detected and measured in a flame ionization
detector.
Reagents:
1. 2,4-D acid standard of known % purity
2. Silvex acid standard of known % purity
3. Acetone, pesticide or spectro grade
4. Ethyl ether, pesticide or spectro grade
5. Dibutyl phthalate, technical (or better)
6. Anhydrous sodium sulfate, ACS granular
7. N-methyl-N-trimethylsilyl-2,2,2-trifluoroacetamide (Eastman 11732):
referred to in this method as MSTFA
8. Internal standard solution - weigh 0.625 gram dibutyl phthalate
into a 500 ml volumetric flask; dissolve in and make to volume
with acetone, (cone 1.25 mg dibutyl phthalate/ml)
-------
Chlorophenoxy Herbicides EPA-5
(Tentative)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 4 ntm ID glass column packed with 10% OV-1 on
60/80 Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 10 ul
4. Soxhlet or Goldfisch extraction apparatus
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: Program from 210° to 245°C at 4°/minute,
hold 4 minutes at final temp, of 245°C
Injection temperature: 250°C
Detector temperature: 250°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi, adjust for particular GC
Hydrogen pressure: 20 psi, adjust for particular GC
Air pressure: 30 psi, adjust for particular GC
Chart speed: 0.25 inches/minute
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure;
Preparation of Standard:
Weigh 0.15 gram 2,4-D acid standard and 0.075 gram silvex
acid standard into a 125 ml glass-stoppered flask or screw-cap
-------
Chlorophenoxy Herbicides EPA-5
(Tentative)
bottle, add 100 ml internal standard solution, and shake to dissolve.
(cone 1.5 mg 2,4-D acid, 0.75 rag silvex acid, and 1.25 mg dibutyl
phthalate/ml)
Preparation of Sample;
Extract 3.75 grams of sample for 1% 2,4-D and 0.5% silvex (or
the equivalent for other % formulations) in a soxhlet or Goldfisch
apparatus for 4-5 hours with ethyl ether. Evaporate the ether on
a steam bath aided by a gentle stream of dry air. Dissolve the
residue in 25 ml internal standard solution and dry with a little
anhydrous sodium sulfate. (cone 1.5 mg 2,4-D acid, 0.75 mg silvex
acid, and 1.25 mg dibutyl phthalate/ml)
Determinat ion;
Injections are made with the syringe filled as follows: 0.5 pi
acetone, 0.5 pi air, 1,0 pi MSTFA, 0.5 pi air, 2 pi of either
standard or sample. Inject 2 pi of standard as above and, if
necessary, adjust the instrument parameters and the volume injected
to give a complete separation within a reasonable time and peak
heights of from 1/2 to 3/4 full scale. The elution order is 2,4-D,
silvex, dibutyl phthalate.
Proceed with the determination, making at least three injections
each of standard and sample solutions using the above mixture for
the injections.
Calculation;
Measure the peak heights or areas of 2,4-D, silvex, and dibutyl
phthalate for both the standard-internal standard solution and the
sample-internal standard solution.
-------
Chlorophenoxy Herbicides EPA-5
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
IS = internal standard = dibutyl phthalate
CPH = chlorophenoxy herbicide
(wt. IS)(% purity IS)(pk. ht. or area CPH)
(wt. CPH)(% purity CPH)(pk. ht. or area IS)
Determine the percent CPH for each injection of the sample-
internal standard solution as follows and calculate the average:
% « (wt. IS)(% purity IS)(pk. ht. or area CPH)
(wt. sample)(pk. ht. or area IS)(RF)
Method submitted by Division of Regulatory Services, Kentucky
Agricultural Experiment Station, University of Kentucky, Lexington,
Kentucky 40506. Data and information on this method and other chloro-
phenoxy herbicides will be appreciated by the editorial committee.
-------
August 1975 Chlorothalonil EPA-1
Determination of Chlorothalonil
by Infrared Spectroscopy
Chlorothalonil is the common name for tetrachloroisophthalonitrile,
a registered fungicide having the chemical structure:
C=N
C
Molecular formula: CgCl.N
Molecular weight: 266
Melting point: 250 to 251°C
Physical state, color, and odor: white crystalline solid, odorless
in pure form; the technical product (about 98%
pure) has a slightly pungent odor.
Solubility: Insoluble in water (0.6 ppm); slightly soluble in
acetone (2% w/w), cyclohexanone (3% w/w), methyl ethyl
ketone (2% w/w), xylene (8% w/w), and kerosene less
than 1%
Stability: stable to ultraviolet radiation and to moderately alkaline
and acid aqueous media; thermally stable under normal
storage conditions; non-corrosive
Other names: Daconil 2787 (Diamond Shamrock Chera. Co.); Bravo; Termil;
2,4,5,6-tetrachloro-l,3-dicyanobenzene; 2,4,5,6-tetra-
chloro-3-cyanobenzonitrile
-------
2 Chlorothalonil EPA-1
Reagents;
1. Chlorothalonil standard of known % purity
2. Methylene chloride, pesticide or spectre grade
3. Sodium sulfate, anhydrous, granular
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.5 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.1 gram Chlorothalonil standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml methylene chloride
by pipette, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 10 mg/ml)
Preparation of Sample:
For emulsifiable concentrates, weigh a portion of sample
equivalent to 0.5 gram Chlorothalonil into a glass-stoppered
flask or screw-cap bottle. Add 50 ml methylene chloride by
-------
3 Chlorothalonil EPA-1
pipette and 1-2 grams anhydrous sodium sulfate. Close tightly
and shake for one hour. Allow to settle; centrifuge or filter
if necessary, taking precaution to prevent evaporation, (final
cone 10 mg chlorothalonil/ml) For low percent formulations
requiring large samples, use more solvent and evaporate an
aliquot to a smaller volume to give a concentration close to
10 mg chlorothalonil/ml.
For flowable formulations, weigh a portion of sample
equivalent to 0.5 gram chlorothalonil into a glass-stoppered
flask or screw-cap bottle. Add 50 ml methylene chloride by
pipette and sufficient anhydrous sodium sulfate to dry and
clarify the methylene chloride solution, (final cone 10 mg
chlorothalonil/ml)
Determination:
With methylene chloride in the reference cell, and using
the optimum quantitative analytical settings for the particular
IR instrument being used, scan both the standard and sample from
1050 cm"1 to 900 cm'1 (9.5 p to 11.1 ji).
Determine the absorbance of standard and sample using the
peak near 980 cm" (10.2 p) and a baseline from 1000 cm to
940 cm"1 (10 fi to 10.64 ja) .
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent chlorothalonil as
follows:
'/ - (abs. sample) (cone, std in mg/ml) (% purity)
(abs. std)(cone, sample in mg/ml)
This method is based on an IR E.G. method submitted by the Commonwealth
of Virginia, Division of Laboratory Services. It should be considered
a tentative method. Any criticism, modification, or verification will
be appreciated.
-------
January 1976 Chloro-Triazine Herbicides EPA-1
Determination of Chloro-Triazine Herbicides
by Chlorine Potentiometric Titration
Several chlorine-containing triazine derivative compounds are
registered as herbicides. They are of the general chemical structure:
ci
R,
The R- and R_ groups are ethylamino, diethylamino, or isopropylamino.
As a group, these compounds generally are: white, crystalline solids;
practically insoluble in water, soluble in organic solvents; stable in
neutral or slightly acidic or basic media, but hydrolyzed by alkali or
mineral acid at higher temperatures; stable to light and heat; and
compatible with most other pesticides.
Principle of the Method;
A potentiometric titration with silver nitrate is used to determine
the total ionic chloride. This includes the chloride liberated from the
triazine by treatment with morpholine and any inorganic chloride present
in the sample. The inorganic chlorine is subtracted from the total
chlorine and the resulting organic chlorine is calculated as the chloro-
triazine herbicide using the appropriate factor for the particular herbi-
cide claimed.
-------
2 Chloro-Triazine Herbicides EPA-1
Reagents;
1. Morpholine
2. Sulfuric acid, 1+4 solution
3. Methyl red indicator
4. Silver nitrate, 0.1N standard solution
5. Ethanol, 95%
6. Sodium or potassium chloride, 0.1N standard solution (exact
normality need not be known if the same volume is titrated
as is added to sample)
Equipment:
1. Potentiometric titrimeter with a silver electrode and a
silver-silver chloride electrode
2. Steam bath
3. Usual laboratory glassware
Procedure:
Determination of Total Chlorine:
Weigh a portion of sample equivalent to 0.4-0.5 gram of the
chloro-triazine derivative into a 125 ml Erlenmeyer flask. Add
20 ml morpholine and heat on the steam bath at full heat for at
least 30 minutes with frequent shaking. Transfer to a 250 ml
beaker with water, acidify with 1+4 sulfuric acid solution using
methyl red as indicator, and cool to room temperature. Titrate
potentiometrically with 0.1N silver nitrate standard solution.
Calculate the total chloride as follows:
% Total chloride = 0*1 AgN03)(N AgN03)(.03545)(100)
(grams sample)
-------
3 Chloro-Triazine Herbicides EPA-1
Determination of Inorganic Chloride:
Weigh a portion of sample equivalent to 0.4-0.5 gram of the
chloro-triazine derivative into a 250 ml beaker. Add 20 ml
ethanol, 150 ml water, and exactly 10 ml of the standard chloride
solution. Acidify with 1+4 sulfuric acid solution using methyl
red as indicator. Titrate potentiometrically with the 0.1N silver
nitrate solution.
Titrate exactly 10 ml of the standard chloride solution as
above except for the sample. Subtract the volume of silver nitrate
used for the standard chloride solution alone from the volume of
silver nitrate used for the sample plus the added standard chloride
solution.
Calculate the inorganic chloride as follows:
% Inorganic chloride = (net ml AgNO^(N AgN03)(.03545)(100)
(grams sample)
Determination of Organic Chloride;
The percent organic chloride is found by subtracting the per-
cent inorganic chloride from the percent total chloride.
% Organic chloride ** % Total chloride - % Inorganic chloride
Calculation of the Chloro-Triazine Herbicide:
The percent chloro-triazine derivative herbicide in the sample
is determined by multiplying the % inorganic chloride by the
appropriate factor for converting chloride to compound.
% Triazine herbicide = % Organic chloride X factor (Cl to cmpd.)
The factors for several chloro-triazine herbicides are as follows:
Anilazine 6.784
Atrazine 6.085
Cyanazine (2 Cl) 3.886
Propazine 6.480
Simazine 5.690
-------
September 1975 Chloroxuron EPA-1
(Tentative)
Determination of Chloroxuron in Dust
by Infrared Spectroscopy
Chloroxuron is the accepted common name for 3-(p-(p-chlorophenoxy)
phenyl)-l,l-dimethylurea, a registered herbicide having the chemical
structure:
Molecular formula: ci5Hi5C1N2°2
Molecular weight: 290.7
Melting point: 149 to 150°C
Physical state, color, and odor: crystalline, white, odorless solid
Solubility: about 3 ppm in water; slightly soluble in ethanol or
benzene; very soluble in acetone or chloroform
Stability: stable; non-corrosive; subject to decomposition by UV
Other names: Tenoran (Ciba-Geigy), Norex, Nor-Am, C-1983,
Chloroxifenid im
Reagents;
1. Chloroxuron standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
Chloroxuron EPA-1
(Tentative)
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.5 mm KBr or NaCl cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.12 gram chloroxuron standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml chloroform by
pipette, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 12 mg/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 1.2 grams chloroxuron
into a glass-stoppered flask or screw-cap bottle. Add 100 ml
chloroform by pipette and 1-2 grams anhydrous sodium sulfate.
Close tightly and shake for one hour. Allow to settle; centri-
fuge or filter if necessary, taking precaution to prevent
evaporation. (final cone 12 mg chloroxuron/ml)
Determination;
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR being used,
scan both the standard and sample from 1430 cm" to 1250 cm"
(7.0 u to 8.0 ju).
Determine the absorbance of standard and sample using the
peak at 1351 cm" (7.40 ji) and basepoint at 1316 cm (7.60^).
-------
Chloroxuron EPA-1
(Tentative)
Calculation:
From the above absorbances and using the standard and
sample solution concentrations, calculate the percent
chloroxuron as follows:
sample>(conc. std in mg/ml)(% purity std)
(abs. std) (cone, sample in mg/ml)
Method submitted by Eva Santos, EPA Product Analysis Laboratory,
San Francisco, California.
-------
October 1975
Chloroxuron EPA-2
(Tentative)
Determination of Chloroxuron
by Gas-Liquid Chromatography
(TCD - Internal Standard)
Chloroxuron is the accepted common name for 3-(p-(p-chlorophenoxy)
phenyl)-l,l-dimethylurea, a registered herbicide having the chemical
structure:
Molecular formula: C H C1N 0
Molecular weight: 290.7
Melting point: 149 to 150°C
Physical state, color, and odor: crystalline, white, odorless solid
Solubility: about 3 ppm in water; slightly soluble in ethanol or
benzene; very soluble in acetone or chloroform
Stability: stable; non-corrosive; subject to decomposition by UV
Other names: Tenoran (Ciba-Geigy), Norex, Nor-Am, C-1983, Chloroxifenidim
Reagents:
1. Chloroxuron standard of known % purity
2. Dieldrin standard of known HEOD content
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.5 gram dieldrin into a
50 ml volumetric flask; dissolve in and make to volume with
acetone. (cone 10 mg dieldrin/ml)
-------
2 Chloroxuron EPA-2
(Tentative)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 4' x 1/8" stainless steel packed with 10% SE-30
on Chromosorb W AW DMCS (or equivalent column)
3. Precision liquid syringe: 25 ul
4. Usual laboratory glassware
Operating Conditions for TCD:
Column temperature: 170°C
Injection temperature: 200°C
Detector temperature: 200°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas pressure: 40 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.08 gram chloroxuron standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 20 ml of
the internal standard solution and shake to dissolve. (final
cone 4 mg chloroxuron and 10 mg dieldrin/ml)
-------
3 Chloroxuron EPA-2
(Tentative)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.08 gram chloroxuron
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the chloroxuron.
For coarse or granular materials, shake mechanically for 10-15
minutes or shake by hand intermittently for 25-30 minutes.
(final cone 4 mg chloroxuron and 10 mg dieldrin/ml)
Determination:
Inject 10-20 ill of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within approx. 15 minutes and peak heights of from
1/2 to 3/4 full scale. The elution time of chloroxuron is approx.
3.5 minutes and that of dieldrin approx. 9 minutes.
Proceed with the determination, making at least three
injections each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of chloroxuron and dieldrin
from both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
_ (wt. dieldrin)(% purity dieldrin)(pk. ht. or area chloroxuron)
(wt. chloroxuron)(% purity chloroxuron)(pk. ht. or area dieldrin)
Determine the percent chloroxuron for each injection of the
sample-internal standard solution as follows and calculate the average:
„ _ (wt. dieldrin)(% purity dieldrin)(pk. ht. or area chloroxuron)
(wt. sample)(pk. ht. or area dieldrin)(RF)
This method is based on EPA, Office of Pesticide Programs, Technical
Services Division, Experimental Method No. 15A. The original source
is unknown and some changes have been made in this write-up; therefore,
any comments, criticisms, suggestions, data, etc. concerning this method
will be appreciated.
-------
November 1975
Coumafuryl EPA-1
Determination of Coumafuryl in Baits
by Ultraviolet Spectroscopy
Coumafuryl is a common name, for 3-(alpha-acetonylfurfuryl)-4-
hydroxycoumarin, a registered rodenticide having the chemical structure:
OH
c=o
Molecular formula: C,,H,.0_
17 14 5
Molecular weight: 298.3
Melting point: pure 122-124°C; technical 119-120°C
Physical state and color: white to tan crystalline solid
Solubility: slightly soluble in cold water; soluble in benzene,
chloroform, methanol, ethanol, isopropanol, ethylene
dichloride, toluene; soluble in most inorganic and organic
bases to form salts
Stability: stable under normal conditions
Other names: Fumarin (Amchem), fumarin (Great Britain, New Zealand),
tomarin (Turkey)
This method is for determining Coumafuryl in most bait materials
and is especially useful for glaze-like coated baits and pellets con-
taining about 0.025% coumafuryl.
-------
2 Coumafuryl EPA-1
Reagents:
1. Coumafuryl standard of known % purity
2. Sodium pyrophosphate, 1% solution - dissolve 5 grams
Na.P-O-.lO H_0 in water and make to 500 ml.
3. Ethyl ether-petroleum ether (20-80) - extract 200 ml petroleum
ether three times with 20 ml portions of pyrophosphate solution
and add 50 ml ethyl ether.
4. Hydrochloric acid, 2.5N solution
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Centrifuge with 100 ml glass-stoppered centrifuge tubes
4. Aspirator or suction device with fine tip glass tube
5. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.1 gram coumafuryl standard into a 100 ml volumetric
flask; dissolve and make to volume with 1% sodium pyrophosphate
solution. Mix thoroughly and pipette 5 ml into a 50 ml volumetric
flask. Make to volume with pyrophosphate solution, mix well,
pipette 5 ml into a second 50 ml volumetric flask, and make to
volume with pyrophosphate solution. (final cone 10 tag/ml)
Preparation of Sample:
Weigh an amount of finely ground sample equivalent to 0.0005
gram coumafuryl (2 grams of 0.025% product) into a 125 ml glass-
stoppered flask, add by pipette 50 ml 1% sodium pyrophosphate
-------
3 Coumafuryl EPA-1
solution, and shake on a mechanical shaker for one hour. Transfer
30-40 ml to a glass-stoppered centrifuge tuhe and centrifuge for
at least 5 minutes. Pipette 25 ml of this solution into a clean
dry 100 ml centrifuge tube. Add 5 ml 2.5N hydrochloric acid and
by pipette 50 ml of the mixed ether solution. Shake for five
minutes. If an emulsion forms, centrifuge to break the emulsion.
Pipette 20 ml of the ether layer to a clean centrifuge tube and
add 10 ml pyrophosphate solution by pipette. Shake for 2 minutes
and remove the ether layer using an aspirator with a glass tube
drawn to a fine tip. If the aqueous layer is not clear, centrifuge
for a few minutes with the stopper off to remove any residual
ether. (final cone 10 jig coumafuryl/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
settings for the particular instrument being used, balance the
pen for 0 and 100% at 305 nm with 1% pyrophosphate solution in
each cell. Scan both standard and sample from 350 nm to 250 nm
with the pyrophosphate solution in the reference cell.
Calculation;
Measure the absorbance of standard and sample at 305 nm and
calculate the percent coumafuryl as follows:
„ = (abs. sample)(cone, std in jig/ml)(% purity std)
(abs. std) (cone, sample in jig/ml)
or using dilution factors, as follows:
(abs. sample)(wt. std)(purity std)(1/100)(5/50)(5/50)(100)
= (abs. std)(wt. sample) (1/50)<25/50)CO/10)
-------
November 1975
Coumafuryl EPA-2
Determination of Coumafuryl in Concentrates
by Ultraviolet Spectroscopy
Coumafuryl is a common name for 3-(alpha-acetonylfurfuryl)-4-
hydroxycoumarin, a registered rodenticide having the chemical structure:
OH
Molecular formula: C,-.H, .0.
17 14 5
c=o
CH3
Molecular weight: 298.3
Melting point: pure 122-124°C; technical 119-120°C
Physical state and color: white to tan crystalline solid
Solubility: slightly soluble in cold water; soluble in benzene, chloro-
form, methanol, ethanol, isopropanol, ethylene dichloride,
toluene; soluble in most inorganic and organic bases to
form salts
Stability: stable under normal conditions
Other names: Fumarin (Amchem), fumarin (Great Britain, New Zealand),
tomarin (Turkey)
This method is for determining coumafuryl in powders containing
about 0.5% coumafuryl.
-------
2 Coumafuryl EPA-2
Reagents;
1. Coumafuryl standard of known % purity
2. Sodium pyrophosphate, 1% solution - dissolve 5 grams
Na.P_0-.10 H00 in water and make to 500 ml.
427 L
3. Ethyl ether, ACS
4. Petroleum ether - extract 200 ml petroleum ether three times
with 20 ml of 1% sodium pyrophosphate solution.
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Centrifuge with 16 x 150 mm glass-stoppered tubes
4. Aspirator or suction device with fine tip glass tube
5. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.1 gram coumafuryl standard into a 100 ml volumetric
flask; dissolve and make to volume with 1% sodium pyrophosphate
solution. Mix thoroughly and pipette 5 ml into a 50 ml volumetric
flask. Make to volume with pyrophosphate solution, mix well,
pipette 5 ml into a second 50 ml volumetric flask, and make to
volume with pyrophosphate solution. (final cone 10 ug/ml)
Preparation of Sample:
Weigh an amount of sample equivalent to 0.0025 gram coumafuryl
(0.5 gram of 0.5% product) into a 125 ml glass-stoppered flask,
add 50 ml ethyl ether by pipette, and shake on a mechanical shaker
-------
3 Coumafuryl EPA-2
for at least 30 minutes. If necessary, centrifuge a portion to
clarify. Pipette 2 ml of the clear ether solution into a 16 x
150 mm glass-stoppered centrifuge tube. Add 10 ml of 1% sodium
pyrophosphate solution by pipette, shake vigorously for two
minutes, and centrifuge at high speed until the aqueous layer is
clear. Draw off the ether layer and any remaining emulsion using
an aspirator with a glass tube drawn into a fine tip. Add 2 ml
ethyl ether, shake, centrifuge, and draw off the ether. Repeat
twice more with 2 ml portions of petroleum ether. If the aqueous
layer is not clear, centrifuge for a few minutes with the stopper
off to remove any residual ether. (final cone 10 pg coumafuryl/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
settings for the particular instrument being used, balance the pen
for 0 and 100% at 305 nm with 1% pyrophosphate solution in each
cell. Scan both standard and sample from 350 nm to 250 nm with
the pyrophosphate solution in the reference cell.
Calculation:
Measure the absorbance of standard and sample at 305 nm and
calculate the percent couraafuryl as follows:
7 = (abs. sample)(cone, std in jug/ml)(% purity std)
(abs. std)(cone, sample in jig/ml)
or using dilution factors, as follows:
7 = (abs. sample)(wt. std)(purity std)(1/100)(5/50)(5/50)(100)
= (abs. std)(wt. sample)(1/50)(2/10)
-------
September 1975
Coumaphos EPA-1
(Tentative)
Determination of Coumaphos
by Infrared Spectroscopy
Coumaphos is the common name for 0,0-diethyl 0-(3-chloro-4-
methyl-2-oxo-2H-l-benzopyran-7-yl) phosphorothioate, a registered
insecticide having the chemical structure:
CH3—CH2—0
«2
\!
p—o
CH3 — CH2—0'
CH3
Molecular formula: C,,H,.C10rPS
14 16 5
Molecular weight: 362.8
Melting point: pure - 95°C; technical - 91 to 92°C
Physical state and color: pure - colorless crystalline solid;
technical - tan or brownish crystalline solid
Solubility: insoluble in water (1.5 ppm at RT); soluble in aromatic
solvents, less so in alcohols and ketones
Stability: hydrolyzes slowly under alkaline conditions; incompatible
with piperonyl butoxide
Other names: Co-Ral (Chemagro), Resitox (Bayer), Asuntol, Baymix,
Meldane, Muscatox, Bay 21/199
-------
2 Coumaphos EPA-1
(Tentative)
Reagents;
1. Coumaphos standard of known % purity
2. Acetone, pesticide or spectro grade
3. Carbon disulfide, pesticide or spectro grade
4. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm KBr or NaCl cells
2. Mechanical shaker
3. Soxhlet extraction apparatus
4. Centrifuge or filtration apparatus
5. Rotary evaporator
6. Cotton or glass wool
7. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.10 gram coumaphos standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml carbon disulfide
by pipette, and shake to dissolve. Add a small amount of
anhydrous sodium sulfate to insure dryness. (final cone 10 mg/ml)
Preparation of Sample;
For high percent formulations (more than 10%), weigh a portion
of sample equivalent to 0.5 gram coumaphos into a glass-stoppered
flask or screw-cap bottle. Add 50 ml carbon disulfide by pipette
-------
Coumaphos EPA-1
(Tentative)
and 1-2 grams anhydrous sodium sulfate. Close tightly and shake
for one hour. Allow to settle; centrifuge or filter if necessary,
taking precaution to prevent evaporation. (final cone 10 mg
coumaphos/ml)
For low percent (less than 10%) formulations, weigh a
portion of sample equivalent to 0.5 gram coumaphos into a
Soxhlet extraction thimble, plug with cotton or glass wool,
and extract with acetone for three hours. Evaporate the acetone
completely on a rotary evaporator. Dissolve the residue, trans-
fer to a 50 ml volumetric flask, and make to volume with carbon
disulfide. Add a small amount of anhydrous sodium sulfate to
clarify and dry the solution. (final cone 10 mg coumaphos/ml)
Determination;
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan both the standard and sample from
1430 cm"1 to 1110 cm"1 (7 p-9 >i) .
Determine the absorbance of standard and sample using the
peak at 1277 cm (7.83 ;j) and baseline from 1307 cm to
1227 cm"1 (7.65 ^ to 8.15 >a).
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent coumaphos as follows:
„ _ (abs. sample)(cone, std in mg/ml)(% purity std)
' __f, —' — ' >r*
(abs. std)(cone, sample in mg/ml)
-------
September 1975
Coumaphos EPA-2
(Tentative)
Determination of Coumaphos
by High Pressure Liquid Chromatography
Coumaphos is the common name for 0,0-diethyl O-(3-chloro-4-
methyl-2-oxo-2H-l-benzopyran-7-yl) phosphorothioate, a registered
insecticide having the chemical structure:
CH3—CH2—0
_j
\1
P—0
CH3—CH2—0'
Molecular formula: C ,H -CICLPS
Molecular weight: 362.8
Melting point: pure - 95°C; technical - 91 to 92°C
Physical state and color: Pure - colorless crystalline solid;
technical - tan or brownish crystalline solid
Solubility: insoluble in water (1.5 ppm at RT); soluble in aromatic
solvents, less so in alcohols and ketones
Stability: hydrolyzes slowly under alkaline conditions; incompatible
with piperonyl butoxide
Other names: Co-Ral (Chemagro), Resitox (Bayer), Asuntol, Baymix,
Meldane, Muscatox, Bay 21/199
Reagents:
1. Coumaphos standard of known % purity
2. Methanol, ACS
-------
Coumaphos EPA-2
(Tentative)
Equipment:
1. High pressure liquid chromatograph with UV detector at 254 nm.
If a variable wavelength UV detector is available, other
wavelengths may be useful to increase sensitivity or eliminate
interference.
2. Suitable column such as:
a. DuPont ODS Pennaphase, 1 meter x 2.1 mm ID
b. Perkin Elmer ODS Sil-X 11 RP, 1/2 meter x 2.6 mm ID
3. High pressure liquid syringe or sample injection loop
4. Usual laboratory glassware
Operating Conditions:
Mobile phase: 40% methanol + 60% water
Column temperature: 50-55°C
Chart speed: 5 rain/inch or equivalent
Flow rate: 0.5 to 1.5 ml/min (Perkin-Elmer 1/2 meter column)
Pressure: 700 psi (DuPont 1 meter column)
Attenuation: Adjusted
Conditions may have to be varied by the analyst for the specific
instrument being used, column variations, sample composition, etc.
to obtain optimum response and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.05 gram couraaphos standard into a small glass-stoppered
flask or vial, add 10 ml methanol by pipette, dissolve and mix
well. (final cone 5 mg/ml)
-------
Coumaphos EPA-2
(Tentative)
Preparation of Sample:
Weigh an amount of sample equivalent to 0.5 gram coumaphos
into a glass-stoppered flask or vial, add 100 ml methanol by
pipette, and shake thoroughly to dissolve the coumaphos. Allow
any solid matter to settle; filter or centrifuge if necessary.
(final cone 5 mg coumaphos/ml)
Determination:
Alternately inject three 10 ul portions each of standard
and sample solutions. Measure the peak height or peak area
for each peak and calculate the average for both standard and
sample.
Adjustments in attenuation or amount injected may have to
be made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent coumaphos as follows:
~, _ (pk. ht. or area sample)(wt. std injectedl(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA, Beltsville, Md.
-------
October 1975
Coumaphos EPA-3
Determination of Coumaphos by
Gas-Liquid Chromatography
(FID - Internal Standard)
Coumaphos is the common name for 0,0-diethyl 0-(3-chloro-4-
methyl-2-oxo-2H-l-benzopyran-7-yl) phosphorothioate, a registered
insecticide having the chemical structure:
CH3—CH2-
CH3—CH2 — 0'
;R—0
Molecular formula: C H ClO^PS
Molecular weight: 362.8
Melting point: pure - 95°C; technical - 91 to 92°C
Physical state and color: pure - colorless crystalline solid;
technical - tan or brownish crystalline solid
Solubility: insoluble in water (1.5 ppm at RT); soluble in aromatic
solvents, less so in alcohols and ketones
Stability: hydrolyzes slowly under alkaline conditions, incompatible
with piperonyl butoxide
Other names: Co-Ral (Chemagro), Resitox (Bayer), Asuntol, Baymix,
Meldane, Muscatox, Bay 21/199
-------
2 Coumaphos EPA-3
Reagents:
1. Coumaphos standard of known % purity
2. Tetradifon standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.09 gram tetradifon
into a 200 ml volumetric flask, dissolve in, and make to
volume with acetone. (cone 0.45 mg tetradifon/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 5' x 1/8" stainless steel packed with 3% SE-30
on 100/120 mesh Varaport 30 (or equivalent
column such as: 6' x 2 mm ID glass column
packed with 3% OV-1 on 60/80 mesh Gas Chrom Q)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 210-220°C
Injection temperature: 250°C
Detector temperature: 250°C
Carrier gas: Nitrogen
Carrier gas flow rate: 60 ml/min
Hydrogen flow rate: 30 ml/min
Air flow rate: 300 ml/min
-------
3 Coumaphos EPA-3
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.05 gram coumaohos standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 50 ml of
the internal standard solution and shake to dissolve. (final
cone 1 mg coumaphos and 0.45 mg tetradifon/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.05 gram coumaphos
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 50 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the coumaphos. For
coarse or granular materials, shake mechanically for 10-15 minutes
or shake by hand intermittently for 25-30 minutes. (final cone
1 mg coumaphos and 0.45 mg tetradifon/ml)
Determination:
Inject 1-3 jal of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and to give peak heights of
1/2 to 3/4 full scale. The elution order is tetradifon, then
coumaphos.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
-------
Coumaphos EPA-3
Calculation:
Measure the peak heights or areas of coumaphos and tetradifon
from both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. tetradifon)(% purity tetradifon)(pk. ht. or area coumaphos)
(wt. coumaphos)(% purity coumaphos)(pk. ht. or area tetradifon)
Determine the percent coumaphos for each injection of the
sample-internal standard solution as follows and calculate the
average:
<•/ _ (wt. tetradifon) (% purity tetradifon) (pk. ht. or area coumaphos)
(wt. sample) (pk. ht. or area tetradifon) (RF) <-{-I)
Method submitted by Division of Regulatory Services, Kentucky
Agricultural Experiment Station, University of Kentucky, Lexington,
Kentucky 40506.
-------
November 1975
Crufomate EPA-1
(Tentative)
Determination of Crufomate
by Infrared Spectroscopy
Crufomate is the accepted common name for 4-tert-butyl-2-chloro-
phenyl methyl methylphosphoramidate, a registered insecticide and
helminthicide having the chemical structure:
CH3-C-X/
Molecular formula: C H C1NCLP
Molecular weight: 292.1
Melting point: 60°C; technical product b.p. 117 to 118°C at 0.01 mm Hg
Physical state and color: white crystalline solid; technical product
is a yellow oil
Solubility: practically insoluble in water and light petroleum but is
readily soluble in acetone, acetonitrile, benzene, carbon
tetrachloride
Stability: stable at pH 7 or below; incompatible with alkaline pesticides
Other names: Ruelene, Dowco 132 (Dow Chemical Co.); 0-methyl 0-2-chloro-
4-tert-butylphenol N-methylamidophosphate
-------
2 Crufomate EPA-1
(Tentative)
Reagents:
1. Crufomate standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording,
with matched 0.2 mm NaCl or KBr cells
2. Mechanical shaker
3. Rotary evaporator with a 60°C water bath
4. Filtration apparatus or centrifuge
5. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.1 gram crufornate standard into a small flask or vial,
add by pipette 10 ml carbon disulfide, and shake to dissolve. Add
a small amount of granular anhydrous sodium sulfate to insure
dryness. (cone 10 mg crufornate/ml)
Preparation of Sample:
For dust, granules, and wettable powder, weigh a portion of
sample equivalent to 1 gram crufomate into a 250 ml glass-
stoppered Erlenmeyer flask, add by pipette 100 ml carbon disulfide,
stopper, and shake on a mechanical shaker for 1 hour. Allow to
settle; filter or centrifuge if necessary. Add a small amount of
granular anhydrous sodium sulfate to insure dryness. (cone 10 mg
crufomate/ml)
-------
3 Crufomate EPA-1
(Tentative)
For liquid formulations and emulsif iable concentrates, weigh
a portion of sample equivalent to 1 gram crufomate into a 100 ml
volumetric flask, make to volume with carbon disulfide, and mix
thoroughly. (Interference from solvents in the sample can some-
times be removed by evaporation on a rotary evaporator under
vacuum at about 60°C before making to volume.) Add a small
amount of granular anhydrous sodium sulfate to insure dryness and
clarify the solution. (cone 10 mg cruf ornate/ml)
An alternative extraction procedure for liquid formulations
and E.C.'s is to shake a 1 gram sample with 100 ml carbon disulfide
and 25-50 ml water in a sealed bottle or flask for 2 hours on a
shaker. Allow to stand for 15 minutes or longer to permit the
carbon disulfide and water layers to separate. With a syringe,
draw off 20-25 ml of carbon disulfide from the bottom of the
bottle and transfer to small vial. Add anhydrous sodium sulfate
to insure dryness and clarify the solution. (cone 10 mg cruf ornate/ml)
IR Determination :
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan both the standard and sample from
1430 cm" to 900 cm"1 (7.0 p to 11.0 p) .
Determine the absorbance of standard and sample using the peak
at 1042 cm" (9.60 p) and a baseline from 1333 cm" to 1000 cm"
(7.50 ^i to 10 ju).
Calculation:
From the above absorbances, calculate the percent crufomate as
follows :
7 = (abs. sample) (cone, std in mg/ml) (% purity std)
(abs. std) (cone, sample in mg/ml)
This method is adapted from Dow Chemical Company method no. 72733,
September 20, 1965.
-------
November 1975 Crufomate EPA-2
(Tentative)
Determination of Crufomate
by Gas-Liquid Chromatography
(TCD - Internal Standard)
Crufomate is the accepted common name for 4-tert-butyl-2-chloro-
phenyl methyl methylphosphoramidate, a registered insecticide and
helminthicide having the chemical structure:
Molecular formula: C^H
Molecular weight: 292.1
Melting point: 60°C; technical product b.p. 117 to 118°C at 0.01 mm Hg
Physical state and color: white crystalline solid; technical product
is a yellow oil
Solubility: practically insoluble in water and light petroleum but is
readily soluble in acetone, acetonitrile, benzene, carbon
tetrachloride
Stability: stable at pH 7 or below; incompatible with alkaline pesticides
Other names: Ruelene, Dowco 132 (Dow Chemical Co.); 0-methyl 0-2-chloro-
4-tert-butylphenol N-methylamidophosphate
-------
2 Crufomate EPA-2
(Tentative)
Reagents:
1. Crufomate standard of known % purity
2. Dieldrin standard of known HECD content
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.5 gram HEOD into a 25 ml
volumetric flask; dissolve in and make to volume with acetone.
(cone 20 mg HEOD/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 6' x 1/8" stainless steel, packed with 10% SE-30
on Chromosorb W AW DMCS (or equivalent column)
3. Precision liquid syringe: 10 ul
4. Usual laboratory glassware
Operating Conditions for TCP:
Column temperature: 170°C
Injection temperature: 200°C
Detector temperature: 200°C
Filament current: 225°C
Carrier gas: Helium
Carrier gas pressure: 40 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Crufomate EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.1 gram crufornate standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 10 ml of the internal
standard solution and shake to dissolve. (final cone 10 mg
crufornate and 20 mg HEOD/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.1 gram crufornate
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 10 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the crufornate. For
coarse or granular materials, shake mechanically for 30 minutes
or shake by hand intermittently for one hour. (final cone 10 mg
crufornate and 20 mg HEOD/ml)
Determination:
Inject 5 ^il of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is crufornate, then HEOD.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of crufomate and HEOD from
both the standard-internal standard solution and the sample-
internal standard solution.
-------
Cruforaate EPA-2
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. HEOD)(% purity HEOD)(pk. ht. or area crufornate)
(wt. crufornate)(% purity crufornate)(pk. ht. or area HEOD)
Determine the percent crufornate for each injection of the
sample-internal standard solution as follows and calculate the
average:
"/ = (wt. HEOD) (% jpurity HEOD) (pk. ht. or area cruf ornate)
= (wt. sample) (pk. ht. or area HEOD) (RF) >iA
Method submitted by David Persch and George Radan, EPA Region II,
New York, N. Y.
-------
September 1975
Cyanazine EPA-1
Determination of Cyanazine
by Infrared Spectroscopy
Cyanazine is the common name for 2-(4-chloro-6-ethylamino-s-
triazin-2-ylamino)-2-methylpropionitrile, a registered herbicide
having the chemical structure:
Cl
•C
N
N
H
CH3
-N
\,
N
C—C=N
Molecular formula: C_H C1N
Molecular weight: 240.7
Melting point: 166.5 to 167°C
Physical state and color: white crystalline solid
Solubility: at 25°C its solubility is 171 ppm in water, 19.5% in
acetone, 1.5% in benzene, 21% in chloroform, 4.5% in
ethanol, 1.5% in hexane, 21% in methylcyclohexanone
Stability: stable to heat and light, and to hydrolysis in neutral
or slightly acidic or basic media
Other names: Bladex (Shell), Fortrol, SD 15418, WL 19805
Reagents;
1. Cyanazine standard of known % purity
2. Methylene chloride, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
2 Cyanazine EPA-1
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.5 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure;
Preparation of Standard:
Weigh 0.25 gram cyanazine standard into a small glass-
stoppered flask or screw-cap tube, add 10 ml methylene chloride
by pipette, close tightly, and shake to dissolve. Add a small
amount of anhydrous sodium sulfate to insure dryness. (final
cone 25 mg/ml)
Preparation of Sample:
Weigh an amount of sample equivalent to 1.25 gram cyanazine
into a glass-stoppered flask or screw-cap bottle. Add 50 ml
methylene chloride by pipette and 1-2 grams anhydrous sodium
sulfate. Close tightly and shake for one hour. Allow to settle;
centrifuge or filter if necessary, taking precautions to prevent
evaporation. (final cone 25 mg cyanazine/ml)
-------
Cyanazine EPA-1
Determination:
With methylene chloride in the reference cell, and using the
optimum quantitative analytical settings, scan both the standard
and sample from 1090 cm"1 to 930 cm" (9.1^i to 10.8^1).
Determine the absorbance of standard and sample using the
peak at 1060 cm" (9.43 u) and basepoint 955 cm" (10.47 /i).
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent cyanazine as
follows:
7 = (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg cyanazine/ml methylene chloride gives
an absorbance of approx. 0.016 in a .5 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
-------
September 1975 Cycloate EPA-1
(Tentative)
Determination of Cycloate
by Gas-Liquid Chromatography (TCD)
Cycloate is the common name for S-ethyl cyclohexylethylthio-
carbamate, a registered herbicide having the chemical structure:
CH2-CH2
0 ,CH CH2
CH3 CH2 S C N\ CH2—CH2
. CHT
Molecular formula: C,.,H NOS
Molecular weight: 215.A
Boiling point: 145°C at 10 mm Hg
Physical state, color, and odor: colorless liquid with an aromatic odor
Solubility: about 100 ppm in water at RT; miscible with most organic
solvents
Stability: stable; non-corrosive
Other names: Ro-Neet (Stauffer Chem. Co.), Eurex, R-2063
Reagents;
1. Cycloate standard of known % purity
2. Chloroform, pesticide or spectro grade
-------
2 Cycloate EPA-1
(Tentative)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 5' x 1/4" glass column packed with 20% SE-30 on
60/80 Chromosorb W, AW, DMCS (or equivalent column)
3. Precision liquid syringe: 50 ^il
4. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating Conditions for TCD:
Column temperature: 210°C
Injection temperature: 240°C
Detector temperature: 270°C
Carrier gas: Helium
Flow rate: 100 ml/min
Operating conditions for filament current, column temperature,
or gas flow should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.20 gram of cycloate standard into a 10 ml volumetric
flask; dissolve and make to volume with chloroform. (final cone
20 mg/ml)
-------
Cycloate EPA-1
(Tentative)
Preparation of Sample:
For technical material and liquid formulations, weigh a
portion of sample equivalent to 0.20 gram cycloate into a
10 ml volumetric flask, make to volume with chloroform,and
mix thoroughly, (final cone 20 mg cycloate/ml)
For dry formulations, weigh a portion of sample equivalent
to 1.0 gram cycloate into a 125 ml screw-cap flask, add by
pipette 50 ml chloroform, and shake for one hour. Allow to
settle; filter or centrifuge if necessary, taking precautions
to prevent evaporation. (final cone 20 mg cycloate/ml)
Determination;
Using a precision liquid syringe, alternately inject
three 20-40 pi portions each of standard and sample solutions.
Measure the peak height or peak area for each peak and calcu-
late the average for both standard and sample.
Adjustments in attenuation or amount injected may have to
be made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the
percent cycloate as follows:
7 x (pk. ht. or area sample)(wt. std injected)(% purity of std)
" (pk. ht. or area standard)(wt. sample injected)
Method submitted by Evangelina Santos, EPA Region IX, San Francisco,
California.
-------
September 1975 Cycloate EPA-2
(Tentative)
Determination of Cycloate
by Gas-Liquid Chromatography (FID)
Cycloate is the common name for S-ethyl cyclohexylethylthio-
carbamate, a registered herbicide having the chemical structure:
CH2-CH2
0 /H CH2
II /\ /
CH3 CH2 S C — N<^ CH2-CH2
'CH2 CH3
Molecular formula: C H NOS
Molecular weight: 215.4
Boiling point: 145°C at 10 mm Hg
Physical state, color, and odor: colorless liquid with an aromatic odor
Solubility: about 100 ppm in water at RT; miscible with most organic
solvents
Stability: stable; non-corrosive
Other names: Ro-Neet (Stauffer Chem. Co.), Eurex, R-2063
Reagents:
1. Cycloate standard of known % purity
2. Acetone, pesticide or spectro grade
-------
2 Cycloate EPA-2
(Tentative)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x I/A" glass column packed with 3% OV-1 on
80/100 Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 10 ^il
4. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 175°C
Injection temperature: 225°C
Detector temperature: 220°C
Carrier gas: Helium or Nitrogen
Flow rate: 50 ml/min
Operating conditions for column temperature, carrier gas flow,
or hydrogen/air flow rates should be adjusted by the analyst to obtain
optimum response and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.10 gram cycloate standard into a 50 ml volumetric
flask; dissolve and make to volume with acetone. (final cone
2 mg/ral)
-------
Cycloate EPA-2
(Tentative)
Preparation of Sample:
For technical material and liquid formulations, weigh a
portion of sample equivalent to 0.10 gram cycloate into a
50 ml volumetric flask, make to volume with acetone,and mix
thoroughly, (final cone 2 mg cycloate/ml)
For dry formulations, weigh a portion of sample equivalent
to 0.2 gram of butylate into a 125 ml screw-cap flask, add by
pipette 50 ml acetone,and shake for one hour. Allow to settle;
filter or centrifuge if necessary,taking precautions to prevent
evaporation. Pipette 25 ml of the clear solution into a 50 ml
volumetric flask and make to volume with acetone and mix
thoroughly. (final cone 2 mg cycloate/ml)
Determination:
Using a precision liquid syringe, alternately inject three
2-4 fil portions each of standard and sample solutions. Measure
the peak height or peak area for each peak and calculate the
average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent cycloate as follows:
y _ (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method developed by Evangelina Santos, EPA Region IX, San Francisco,
California.
-------
October 1975 Cycloate EPA-3
Determination of Cycloate by
Gas-Liquid Chromatography
(FID - Internal Standard)
Cycloate is the common name for S-ethyl cyclohexylethylthio-
carbamate, a registered herbicide having the chemical structure:
CH2-CH2
0 CH2
0 /H
II /\
- C - N
CH3_CH2 - S - C - N CH2-CH2
- CH3
Molecular formula: C H NOS
Molecular weight: 215.4
Boiling point: 145°C at 10 mm Kg
Physical state, color, and odor: colorless liquid with an aromatic odor
Solubility: about 100 ppm in water at R.T; miscible with most organic
solvents
Stability: stable; non-corrosive
Other names: Ro-Neet (Stauffer Chem. Co.), Eurex, R-2063
Reagents ;
1. Cycloate standard of known % purity
2. Pebulate standard of known % purity
3. Carbon disulfide, pesticide or spectro grade
4. Chloroform, pesticide or spectro grade
-------
2 Cycloate EPA-3
Reagents (Cont.):
5. Methanol, pesticide or spectro grade
6. Internal Standard solution - weigh 0.2 gram pebulate into a
50 ml volumetric flask, dissolve in, and make to volume with
a solvent mixture consisting of 80% carbon disulfide + 15%
chloroform + 5% methanol. (cone 4 mg pebulate/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 4 mm ID glass column packed with 3% OV-1 on
60/80 mesh Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 140°C
Injection temperature: 225°C
Detector temperature: 250°C
Carrier gas: Nitrogen
Carrier gas flow rate: (not stated in the method)
Hydrogen flow rate: (not stated in the method)
Air flow rate: (not stated in the method)
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Cycloate EPA-3
Procedure:
Preparation of Standard:
Weigh 0.08 gram cycloate standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 20 ml of the internal
standard solution and shake to dissolve, (final cone 4 mg
cycloate and 4 mg pebulate/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.08 gram cycloate
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the cycloate. For
coarse or granular materials, shake mechanically for 10-15
minutes or shake by hand intermittently for 25-30 minutes.
(final cone 4 mg cycloate and 4 mg pebulate/ml)
Determination:
Inject 1-2 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is pebulate, then cycloate.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of cycloate and pebulate
from both the standard-internal standard solution and the sample-
internal standard solution.
-------
Cycloate EPA-3
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
_ (wt. pebulate)(% purity pebulate)(pk. ht. or area cycloate)
(wt. cycloate)(% purity cycloate)(pk. ht. or area pebulate)
Determine the percent cycloate for each injection of the sample-
internal standard solution as follows and calculate the average:
7 - (wt- pebulate)(% purity pebulate)(pk. ht. or area cycloate)
(wt. sample) (pk. ht. or area pebulate) (RF) / U - Ij
Method submitted by Division of Regulatory Services, Kentucky
Agricultural Experiment Station, University of Kentucky, Lexington,
Kentucky 40506.
-------
-------
December 1975 Dalapon EPA-1
Determination of Dalapon
by Infrared Spectroscopy
Dalapon is the accepted common name for 2,2-dichloropropionic
acid, a registered herbicide having the chemical structure:
Cl
I
CH3—C-COOH
Cl
Molecular formula: C.H.Cl-CL
34 22
Molecular weight: 143
Boiling point: 185 to 190°C
Physical state, color, and odor: colorless, odorless liquid
Solubility: very soluble in water, ethanol, alkali solvents; soluble
in ether, carbon disulfide
Stability: nonflammable; compatible with hard water and liquid
fertilizers; mildly corrosive; stable in dry form;
sodium and magnesium salts are hygroscopic
Other names: Dowpon, Radapon (Dow Chem. Co.); Basfapon, Ded-Weed,
Gramevin, Unipon
Reagents:
1. 2,2-Dichloropropionic acid (or sodium salt) of known % purity
2. Carbon disulfide, ACS (spectroscopic grade preferred)
3. Sulfuric acid, 1+3
4. Anhydrous sodium sulfate, ACS granular
-------
Dalapon EPA-1
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 1.0 mm NaCl or KBr cells
2. Mechanical shaker, wrist action
3. 4 oz. screw-cap bottles
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.6-0.7 gram 2,2-dichloropropionic acid, or 0.7-0.8 gram
2,2-dichloropropionic acid sodium salt into a 4 oz. screw-cap
bottle. Proceed as under Preparation of Sample, second paragraph,
"Add 2 ml ."
Preparation of Sample:
Weigh a portion of sample equivalent to 0.6-0.7 gram 2,2-
dichloropropionic acid or 0.7-0.8 gram 2,2-dichloropropionic acid
sodium salt into a 4 oz. screw-cap bottle.
Add 2 ml sulfuric acid solution and mix well. By pipette add
100 ml carbon disulfide and shake on a mechanical shaker for 20
minutes. Add sufficient granular anhydrous sodium sulfate to absorb
all the water and clarify the solution. Shake an additional 10
minutes and allow to settle.
Determination;
Using the optimum quantitative analytical settings for the
particular IR spectrophotometer being used, scan the standard and
sample solutions from 1333 cm~ to 910 cm (7.5^i to ll.Oyu) using
carbon disulfide in the reference cell. For qualitative comparison,
run a full scan.
-------
Dalapon EPA-1
Determine the absorbance of both the standard and sample using
the peak at 1130 cm (8.85 ji) and a base line from 1155 cm to
1015 cm"1 (8.65 to 9.85 ji) .
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent dalapon (or dalapon sodium
salt) as follows:
% „ (abs. sample)(wt. std)(purity std)(1/100)(100)
(abs. std)(grams sample)(1/100)
% dalapon sodium salt = (1.1537)(% dalapon)
This method was adapted from The Dow Chemical Company method for Dowpon C.
-------
January 1976
DDT EPA-1
Determination of DDT in Emulsifiable Concentrates
Containing a Volatile Solvent by Infrared Spectroscopy
DDT is a common name for dichlorodiphenyltrichoroethane, an
insecticide having the chemical structure:
p ,p'-isomer data:
Molecular formula: C.,H Cl
Molecular weight: 354.5
Melting point: 108.5eC
Physical state and color: colorless crystals
Solubility: practically insoluble in water; moderately soluble in
hydroxylic and polar solvents such as alcohol, and in
petroleum oils; soluble in most aromatic and chlorinated
solvents
Stability: dehydrochlorinated at temperatures above m.p., a reaction
catalyzed by ferric and aluminum chloride and by UV light;
readily dehydrochlorinated when in solution in organic
solvents by alkali or organic bases; otherwise stable and
inert, unattacked by acid and alkaline permanganate or by
aqueous acids and alkalis
-------
2 DDT EPA-1
Technical: The technical product (up to 30% o,p'-isomer) is a waxy
solid of indefinite m.p. and of similar solubility to the
p,p'-isomer.
Other names: Gesarol, Guesarol, Neocid (Ciba-Geigy); Dicophane (British
Pharmacopeia); chlorophenothane (U.S. Pharmacopoeia);
Zerdane (France); anofex, Dedelo, Didimac, Genitox, Gesapon,
Gesarex, Gyron, Ixodex, Kopsol, Pentachlorin, Rukseam,
1,1,l-trichloro-2,2-bis(p-chlorophenyl)ethane
Reagents:
1. Technical DDT standard
2. Carbon disulfide, ACS
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.5 mm NaCl or KBr cells
2. Rotary evaporator with 60° water bath
3. Usual laboratory glassware
Procedure;
Preparation of Standard;
Weigh 0.4 gram technical DDT into a 25 ml volumetric flask,
dissolve in, and make to volume with carbon disulfide. Add a
small amount of anhydrous sodium sulfate to insure dryness.
(cone 16 mg/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.4 gram technical DDT
into a 125 ml standard tapered Erlenmeyer flask and evaporate the
-------
3 DDT EPA-1
solvent on a rotary evaporator using a water bath at 60°C. The
solvent (e.g., xylene) can usually be evaporated in about 10 minutes,
but the DDT may not crystallize; however, the last traces of sol-
vent may be removed with a gentle stream of air.
Dissolve the residue, transfer quantitatively to a 25 ml volu-
metric flask, and make to volume with carbon disulfide. Add a
small amount of anhydrous sodium sulfate to insure dryness. (cone
16 mg tech. DDT/ml)
Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan the standard and sample from 1175 cm
to 950 cm" (8.5 p. to 10.5 p).
Determine the absorbance of standard and sample using the peak
at 1017 cm (9.83 p.) and baseline from 1064 cm"1 to 970 cm (9.4 p
to 10.3 ji) .
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent technical DDT as follows:
7 = (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
-------
December 1975
Deet EPA-1
(Tentative)
Determination of Deet
by Infrared Spectroscopy
Deet is the common name for N,N-diethyl-m-toluamide, a registered
insect repellent having the chemical formula:
—N'
,CH2-CH3
•CH2-CH3
Molecular formula: C -H NO
Molecular weight: 191.3
Boiling point: 111°C at 1 mm Hg
Physical state and color: colorless to amber liquid, nearly odorless;
the technical product contains 85-95% m isomer;
the o and p isomers are highly repellent but less
effective than the m isomer
Solubility: practically insoluble in water; miscible with ethanol,
isopropanol, propylene glycol, cottonseed oil, ether,
benzene
Stability: stable under normal conditions; non-corrosive to most metals
Other names: Metadelphene (Hercules), Delphene, Detamide, Off
This method is primarily for alcohol solutions.
-------
2 Deet EPA-1
(Tentative)
Reagents;
1. Deet standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotoraeter, double beam ratio recording with
matched 0.5 mm NaCl or KBr cells
2. Rotary evaporator
3. Water bath at 50°C
4. Usual laboratory glassware
Procedure:
Preparation of Standard;
Weigh 0.4 gram deet standard into a 10 ml volumetric flask and
make to volume with carbon disulfide. Add a small amount of
anhydrous sodium sulfate to insvire dryness. (cone 40 mg/ml)
Preparation of Sample:
Weigh a portion of sample (alcohol solution and aerosol non-
volatile) equivalent to 0.4 gram deet into a 125 ml Erlenmeyer
flask and evaporate the alcohol under vacuum on a rotary evaporator
at 50°C. (The alcohol may be removed by heating on a steam bath for
a few minutes with a slow current of air passing into the flask.)
Do not heat any longer than necessary to remove the alcohol. Trans-
fer the residue quantitatively to a 10 ml volumetric flask and make
to volume with carbon disulfide. Add a small amount of anhydrous
sodium sulfate and shake thoroughly to remove water and clarify the
solution. (final cone 40 mg deet/ml)
-------
Deet EPA-1
(Tentative)
De t ermina t ion:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan both the standard and sample from
770 cm"1 to 665 cm"1 (13 p to 15 /u) .
Determine the absorbance of standard and sample using the
peak at 706.7 cm" (14.15jj) and basepoint 692.5 cm" (14.44yu).
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent deet as follows:
7 = (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
The above method is based on the old USDA, PRD Methods
Clearing House Method No. 382.0 and on EPA's Exp. Method 26B and is
for alcohol solutions of the meta isomer.
The para isomer may be determined in a similar manner using
877.2 cm (11.4 u) analytical peak and 862.1 cm (11.6 u) basepoint.
Some success has been obtained by the Beltsville Chemistry Lab-
oratory on aerosols, creams, and sticks: sometimes by extraction from
aqueous mixtures using carbon disulfide, filtering, and drying with
anhydrous sodium sulfate; and sometimes by choosing another IR wave-
length where interferences from sample components (IR scanned) are not
present.
-------
December 1975
Deet EPA-2
(Tentative)
Determination of Deet by
Gas-Liquid Chromatography
(TCD - Internal Standard)
Deet is the common name for N,N-diethyl-m-toluamide, a registered
insect repellent having the chemical formula:
CH2-CH3
CH2-CH3
Molecular formula: C ?H _NO
Molecular weight: 191.3
Boiling point: 111°C at 1 mm Hg
Physical state and color: colorless to amber liquid, nearly odorless;
the technical product contains 85-95% m isomer;
the o and p isomers are highly repellent but less
effective than the m isomer
Solubility: practically insoluble in water; miscible with ethanol,
isopropanol, propylene glycol, cottonseed oil, ether,
benzene
Stability: stable under normal conditions; non-corrosive to most metals
Other names: Metadelphene (Hercules), Delphene, Detamide, Off
This method is for aerosols containing MGK 264, MGK 326, and
MGK Repellent II.
-------
2 Deet EPA-2
(Tentative)
Reagents:
1. Deet standard of known % purity
2. Heptachlor standard of known % purity
3. Benzene, pesticide or spectro grade
4. Internal Standard solution - weigh 1.2 grams heptachlor into
a 100 ml volumetric flask; dissolve in and make to volume
with benzene. (cone 12 mg heptachlor/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 6' x 1/8" O.D. stainless steel, packed with 10% SE-30
on 80/100 Diatoport S (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Usual laboratory glassware
Operating Conditions^ for TCD;
Column temperature: 190°C
Injection temperature: 215°C
Detector temperature: 215°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas flow rate: adjust for specific GC
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Deet EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.08 gram deet standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 20 ml of the internal
standard solution and shake thoroughly. (final cone 4 mg deet
and 12 mg heptachlor/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.08 gram deet into a
small glass-stoppered flask or screw-cap bottle. Add by pipette
20 ml of the internal standard solution. Close tightly and shake
thoroughly. (final cone 4 mg deet and 12 mg heptachlor/ml)
Determinat ion:
Inject 2-3 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is deet, then heptachlor.
Technical heptachlor gives a second small peak which should be
eluted before another injection.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of deet and heptachlor from
both the standard-internal standard solution and the sample-internal
standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
„ = ^wt. hep_t_achj.or) (%_ purity heptachlor)(pk. ht. or area deet)
(wt. deet)(% purity deet)(pk. ht. or area heptachlor)
-------
Deet EPA-2
(Tentative)
Determine the percent deet for each injection of the sample-
internal standard solution as follows and calculate the average:
«„ = (wt. heptachlor) (% purity heptachlor) (jik. ht_. or area deet) (JrQ^)
(wt. sample) (pk. ht. or area heptachlor) (RF) £f~//'
This method is based on EPA's Exp. Method No. 26 submitted by Stelios
Gerazounis, EPA, Region II, New York, N. Y.
Although specifically for aerosol samples, this method with modification
could be used for other deet formulations. Any suggestions, data,
comments, etc. will be appreciated.
-------
December 1975
Deet EPA-3
(Tentative)
Determination of Deet by
Gas-Liquid Chromatography
(FID - Internal Standard)
Deet is the common name for N,N-diethyl-m-toluamide, a registered
insect repellent having the chemical formula:
CH2- CH3
CH2-CH3
Molecular formula: C. _H _NO
Molecular weight: 191.3
Boiling point: 111°C at 1 mm Hg
Physical state and color: colorless to amber liquid, nearly odorless;
the technical product contains 85-95% m isomer;
the o and p isomers are highly repellent but less
effective than the m isomer
Solubility: practically insoluble in water; miscible with ethanol,
isopropanol, propylene glycol, cottonseed oil, ether,
benzene
Stability: stable under normal conditions; non-corrosive to most metals
Other names: Metadelphene (Hercules), Delphene, Detamide, Off
-------
2 Deet EPA-3
(Tentative)
Reagents:
1. Deet standard of known % purity
2. Vernolate standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.2 gram vernolate standard
into a 100 ml volumetric flask, dissolve in, and make to volume
with acetone, (cone 2 mg vernolate/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: A1 x 2 mm ID glass column packed with 3% OV-17 on
80/100 Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 150°C
Injection temperature: 200°C
Detector temperature: 200°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi (adjusted for specific GC)
Hydrogen pressure: 20 psi (adjusted for specific GC)
Air pressure: 30 psi (adjusted for specific GC)
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Deet EPA-3
(Tentative)
Procedure:
Prejjaratiqn of Standard:
Weigh 0.075 gram deet standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 25 ml of the internal
standard solution and shake thoroughly. (final cone 3 mg deet
and 2 mg vernolate/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.075 gram deet into
a small glass-stoppered flask or screw-cap bottle. Add by pipette
25 ml of the internal standard solution. Close tightly and shake
thoroughly. (final cone 3 mg deet and 2 mg vernolate/ml)
Determination:
Inject 1-2 jil of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is vernolate, then deet.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of deet and vernolate from
both the standard-internal standard solution and the sample-internal
standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
RF (wt. vernolate)(% purity vernolate)(pk. ht. or area deet)
(wt. deet)(% purity deet)(pk. ht. or area vernolate)
-------
Deet EPA-3
(Tentative)
Determine the percent deet for each injection of the sample-
internal standard solution as follows and calculate the average:
7 - (wt. vernoj-ate) (% purity vgrnplate) Cpjc^ ht . or area deet)
(wt. sample) (pk. ht. or area vernolate) (RF) LA- I'-
This method was submitted by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
Note! This method has been designated as tentative since it is a Va.
Exp. Tent, method and because some of the data has been suggested
by EPA's Beltsville Chemistry Lab. Any comments, criticisms,
suggestions, data, etc. concerning this method will be appreciated,
especially as related to analysis of different deet formulations.
-------
November 1975
Diazinon EPA-1
Determination of Diazinon
by Gas-Liquid Chromatography (TCD)
Diazinon is the common name (ISO and BSI, except U.S. where it
is a registered trademark) for 0,0-diethyl 0-(2-isopropyl-6-methyl-4-
pyrimidinyl) phosphorothioate; a registered insecticide, acaricide,
and nematocide having the chemical structure:
CH3 CH2 0
CH3 CH2 — 0'
P 0
•N
Molecular formula: Ci2H21N2°3PS
Molecular weight:
Boiling point:
304.3
83 to 8A°C under 0.002 mm Hg
Physical state and color: colorless liquid; the technical product
(about 95% pure) is light amber to dark brown.
Solubility: 0.004% (40 ppm) in water at RT; miscible with aliphatic and
aromatic solvents, alcohols, and ketones; soluble in
petroleum oils
Stability: decomposes about 120°C; susceptible to oxidation; stable
in alkaline media but slowly hydrolyzed in water and
dilute acids; compatible with most pesticides but should
not be combined with copper fungicides; the presence of
traces of water promotes hydrolysis on storage to the
highly poisonous tetraethyl monothiopyrophosphate
-------
2 Diazinon EPA-1
Other names: Spectracide, Diazinon, G-24480 (Ciba-Geigy); Basudin,
Diazajet, Diazide, Diazol, Dazzel, Gardentox, Neocidol,
Nucidol, Sarolex
Reagents;
1. Diazinon standard of known % purity
2. Acetone, pesticide or spectro grade
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 4' x 1/4" O.D. glass column packed with 5% SE-30 on
60/80 mesh Chromosorb W, AW, DMCS (or equivalent
column)
3. Precision liquid syringe: 50 pi
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for TCP;
Column temperature: 170°C
Injection temperature: 200°C
Detector temperature: 200°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas pressure: 30-40 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response and
reproducibility.
-------
3 Diazinon EPA-1
Procedure;
Preparation of Standard:
Weigh 0.1 gram diazinon standard into a 10 ml volumetric flask;
dissolve in and make to volume with acetone. (final cone 10 mg
diazinon/ml)
Preparation of Sample:
For emulsifiable concentrates and liquid formulations, weigh
a portion of sample equivalent to 0.1 gram diazinon into a 10 ml
volumetric flask, make to volume with acetone,and mix thoroughly.
(final cone 10 mg diazinon/ml)
For dry formulations, weigh a portion of sample equivalent
to 0.5 gram diazinon into a 125 ml screw-cap flask, add by
pipette 50 ml acetone, and shake for one hour. Allow to settle;
filter or centrifuge if necessary, taking precautions to prevent
evaporation, (final cone 10 mg diazinon/ml)
Determination:
Using a precision liquid syringe, alternately inject three
15-25 pi portions each of standard and sample solutions. Measure
the peak height or peak area for each peak and calculate the
average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the percent
diazinon as follows:
2 a (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
-------
November 1975
Diazinon EPA-2
(Tentative)
Determination of Diazinon by
High Pressure Liquid Chromatography
Diazinon is the common name (ISO and BSI, except U.S. where it
is a registered trademark) for 0,0-diethyl 0-(2-isopropyl-6-methyl-4-
pyrimidinyl) phosphorothioate; a registered insecticide, acaricide,
and nematocide having the chemical structure:
CH3
0,
CH3 CH2—0'
— 0
Molecular formula: C.-H N 0 PS
Molecular weight:
Boiling point:
304.3
83 to 84°C under 0.002 mm Hg
Physical state and color: colorless liquid; the technical product
(about 95% pure) is light amber to dark brown.
Solubility: 0.004% (40 ppm) in water at RT; miscible with aliphatic and
aromatic solvents, alcohols, and ketones; soluble in
petroleum oils
Stability: decomposes about 120°C; susceptible to oxidation; stable
in alkaline media but slowly hydrolyzed in water and
dilute acids; compatible with most pesticides but should
not be combined with copper fungicides; the presence of
traces of water promotes hydrolysis on storage to the
highly poisonous tetraethyl monothiopyrophosphate
-------
2 Diazinon EPA-2
(Tentative)
Other names: Spectracide, Diazinon, G-24480 (Ciba-Geigy); Basudin,
Diazajet, Diazide, Diazol, Dazzel, Gardentox, Neocidol,
Nucidol, Sarolex
Reagents;
1. Diazinon standard of known % purity
2. Methanol, pesticide or spectro grade
Equipment:
1. High pressure liquid chromatograph with UV detector at 254 nm.
If a variable wavelength UV detector is available, other
wavelengths may be useful to increase sensitivity or eliminate
interference. 235 nm has been found useful for methyl parathion.
2. Suitable column such as:
a. DuPont ODS Pennaphase, 1 meter x 2.1 mm ID
b. Perkin Elmer ODS Sil-X-II RP, 1/2 meter x 2.6 mm ID
3. High pressure liquid syringe or sample injection loop
4. Usual laboratory glassware
Operating Conditions:
Mobile phase: 20% methanol + 80% water
Column temperature: 50-55°C
Chart speed: 5 min/inch or equivalent
Flow rate: 0.5 to 1.5 ml/min (Perkin-Elmer 1/2 meter column)
Pressure: 900 psi (DuPont 1 meter column)
Attenuation: Adjusted
Conditions may have to be varied by the analyst for the specific
instrument being used, column variations, sample composition, etc.
to obtain optimum response and reproducibility.
-------
Diazinon EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.1 gram diazinon standard into a 50 ml volumetric
flask, add 50 ml methanol by pipette, and mix thoroughly.
(final cone 2 mg diazinon/ml)
Preparation of Sample:
Weigh an amount of sample equivalent to 0.1 gram diazinon
into a glass-stoppered flask or vial, add 50 ml methanol by
pipette,and shake thoroughly to dissolve the diazinon. Allow
any solid matter to settle; filter or centrifuge if necessary.
(final cone 2 mg diazinon/ml)
Determination:
Alternately inject three 5 ^il portions each of standard
and sample solutions. Measure the peak height or peak area for
each peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the percent
diazinon as follows:
(pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA, Beltsville, Md.
-------
November 1975
Diazinon EPA-3
Determination of Diazinon
by Infrared Spectroscopy
Diazinon is the common name (ISO and BSI, except U.S. where it
is a registered trademark) for 0,0-diethyl 0-(2-isopropyl-6-methyl-4-
pyrimidinyl) phosphorothioate; a registered insecticide, acaricide,
and nematocide having the chemical structure:
CH3 CH2—0
CH3 CH2—0'
P 0
Molecular formula: c12H2iN2°3PS
Molecular weight: 304.3
Boiling point: 83 to 84°C under 0.002 mm Hg
Physical state and color: colorless liquid; the technical product
(about 95% pure) is light amber to dark brown.
Solubility: 0.004% (40 ppm) in water at RT; miscible with aliphatic and
aromatic solvents, alcohols, and ketones; soluble in
petroleum oils
Stability: decomposes about 120°C; susceptible to oxidation; stable
in alkaline media but slowly hydrolyzed in water and
dilute acids; compatible with most pesticides but should
not be combined with copper fungicides; the presence of
traces of water promotes hydrolysis on storage to the
highly poisonous tetraethyl monothiopyrophosphate
-------
2 Diazinon EPA-3
Other names: Spectracide, Diazinon, G-24480 (Ciba-Geigy); Basudin,
Diazajet, Diazlde, Diazol, Dazzel, Gardentox, Neocidol,
Nucidol, Sarolex
Reagents;
1. Diazinon standard of known % purity
2. Acetone, pesticide or spectro grade
Equipment:
1. Infrared spectrophototneter, double beam ratio recording with
matched 0.2 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, put in 1-2 grams anhydrous sodium sulfate, and seal
tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50 RPM
on a standard Patterson-Kelley twin shell blender that has
been modified by replacing the blending shell with a box to
hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.1 gram diazinon standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml acetone by pipette, close
tightly, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 10 mg diazinon/ml)
-------
3 Diazinon EPA-3
Preparation of Sample:
Weigh an amount of sample equivalent to 0.5 gram diazinon into
a glass-stoppered flask or screw-cap tube. Add 50 ml acetone by
pipette and 1-2 grams anhydrous sodium sulfate. Close tightly and
shake for one hour. Allow to settle centrifuge or filter if
necessary, taking precautions to prevent evaporation, (final cone
10 mg diazinon/ml)
Determination:
With acetone in the reference cell, and using the optimum
quantitative analytical settings, scan both the standard and
—1 —1
sample from 925 cm to 715 cm (10.8 p to 14.0 ji) .
Determine the absorbance of standard and sample using the
peak at 833.3 cm" (12.0 u) and a baseline from 719.4 cm~ to
1123.6 cm"1 (13.9 p to 8.9 yu) .
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent diazinon as follows:
2 = (abs. sample)(cone. std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg diazinon/ml acetone gives an absorbance
of approx. 0.033 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond, Va. 23219.
-------
November 1975
Diazlnon EPA-4
Determination of Diazinon
by Gas-Liquid Chromatography
(FID - Internal Standard)
Diazinon is the common name (ISO and BSI, except U.S. where it
is a registered trademark) for 0,0-diethyl 0-(2-isopropyl-6-methyl-4-
pyrimidinyl) phosphorothioate; a registered insecticide, acaricide,
and nematocide having the chemical structure:
CH3 CH2 0
CH3 CH2 0'
P 0
CH3
Molecular formula: C12H21N2°3PS
Molecular weight: 304.3
Boiling point: 83 to 84°C under 0.002 mm Hg
Physical state and color: colorless liquid; the technical product
(about 95% pure) is light amber to dark brown.
Solubility: 0.004% (40 ppm) in water at RT; miscible with aliphatic and
aromatic solvents, alcohols, and ketones; soluble in
petroleum oils
Stability: decomposes about 120°C; susceptible to oxidation; stable
in alkaline media but slowly hydrolyzed in water and
dilute acids; compatible with most pesticides but should
not be combined with copper fungicides; the presence of
traces of water promotes hydrolysis on storage to the
highly poisonous tetraethyl monothiopyrophosphate
-------
2 Diazinon EPA-4
Other names: Spectracide, Diazinon, G-2448Q (Ciba-Geigy); Basudin,
Diazajet, Dlazlde, Diazol, Dazzel, Gardentox, Neocldol,
Nucidol, Sarolex
Reagents;
1. Diazinon standard of known % purity
2. Aldrin of known HHDN content
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.150 gram HHDN into a
50 ml volumetric flask, dissolve in, and make to volume with
acetone. (cone 3 mg HHDN/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 4 mm ID glass column packed with 3% OV-1 on
60/80 mesh Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 jil
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 175°C
Injection temperature: 260°C
Detector temperature: 255°C
Carrier gas: Nitrogen
Carrier gas pressure: (not stated in method) (40-60 psi)
Hydrogen pressure: 20 psi
Air pressure: 30 psi
-------
3 Diazinon EPA-4
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response and
reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.04 gram diazinon standard into a small glass~stoppered
flask or screw-cap bottle. Add by pipette 20 ml of the internal
standard solution and shake to dissolve. (final cone 2 mg diazinon
and 3 mg HHDN/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.04 gram diazinon into
a small glass-stoppered flask or screw-cap bottle. Add by pipette
20 ml of the internal standard solution. Close tightly and shake
thoroughly to dissolve and extract the diazinon. For coarse or
granular materials, shake mechanically for 30 minutes or shake by
hand intermittently for one hour. (final cone 2 mg diazinon and
3 mg HHDN/ml)
Determinat ion:
Inject 2-3 ul of standard and, if necessary, adjust the instru-
ment parameters and the volume injected to give a complete separation
within a reasonable time and peak heights of from 1/2 to 3/4 full
scale. The elution order is diazinon, then HHDN.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of diazinon and HHDN from both
the standard-internal standard solution and the sample-internal
standard solution.
-------
DIazinon EPA-4
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
m (wt. HHDN)(% purity HHDN)(pk. ht. or area diazinon)
(wt. diazinon)(% purity diazinon)(pk. ht. or area HHDN)
Determine the percent diazinon for each injection of the
sample-internal standard solution as follows and calculate the
average:
v = (wt. HHDN)(% purity HHDN)(pk. ht. or area diazinon)
(wt. sample) (pk. ht. or area HHDN) (RF)
Method submitted by Division of Regulatory Services, Kentucky Agricul-
tural Experiment Station, University of Kentucky, Lexington, Kentucky 40506.
-------
November 1975
Dibromochloropropane EPA-1
Determination of Dibromochloropropane
by Infrared Spectroscony
Dibromochloropropane is the trivial name for l,2-dibromo-3-chloro-
propane, a registered soil fumigant and nenatocide having the chemical
structure:
H-
H
H
H
Br Br Cl
H
Molecular formula: C H Br Cl
Molecular weight: 236.3
Boiling point: 196°C
Physical state, color, and odor: amber to dark brown dense liquid
with a mildly pungent odor
Solubility: 1000 ppm in water; miscible with aliphatic and aromatic
solvents
Stability: stable to hydrolysis in neutral or acid media; hydrolyzed
by alkali to 2-bromoallyl alcohol; corrodes aluminum,
magnesium, and tin alloys
Other names: Fumazone (Dow Chemical Co.), Nemagon (Shell Development Co.),
DBCP, Nemafume, BBC 12, OS 1897
-------
2 Dibromochloropropane EPA-1
Reagents:
1. Dibromochloropropane standard of known % purity
2. Acetone, pesticide or spectro grade
3. Carbon disulfide, pesticide or spectro grade
4. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.2 mm KBr cells
A
2. Mechanical shaker
3. Rotary evaporator
4. Centrifuge or filtration apparatus
*
5. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, put in 1-2 grams anhydrous sodium sulfate, and
seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50 RPM
on a standard Patterson-Kelley twin shell blender that has been
modified by replacing the blending shell with a box to hold a
24-tube rubber-covered rack.
Procedure:
Preparation of Standard^:
Weigh 0.5 gram dibromochloropropane into a 10 ml volumetric
flask, dissolve in, and make to volume with carbon disulfide. Add
a small amount of anhydrous sodium sulfate to insure dryness.
(final cone 50 mg/ml)
-------
3 Dibromochloropropane EPA-1
Preparation of Sample:
For emulsifiable concentrates, weigh a portion of sample equiv-
alent to 0.5 gram dibromochloropropane into a 10 ml volumetric flask,
make to volume with carbon disulfide, and mix well. Add a small
amount of anhydrous sodium sulfate to insure dryness. (final cone
50 mg dibromochloropropane/ml)
For granular formulations, weigh a portion of sample equivalent
to 1.0 gram dibromochloropropane into a glass-stoppered flask or
screw-cap tube. Add 50 ml acetone by pipette and 1-2 grams anhydrous
sulfate. Close tightly and shake for one hour. Allow to settle;
centrifuge or filter if necessary, taking precaution to prevent
evaporation. Evaporate a 25 ml aliquot to dryness on a water bath
using a gentle stream of dry air; evaporate the last one or two ml
with air only. Dissolve in about 4-5 ml carbon disulfide, transfer
to a 10 ml volumetric flask, and make to volume with carbon disulfide.
Add a small amount of anhydrous sodium sulfate to insure drvness.
(final cone 50 mg dibromochloropropane/ml)
Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
_i
instrument being used, scan the standard and sample from 800 cm
to 500 cm"1 (12.5 p to 20 p).
Determine the absorbance of standard and sample using the peak
-1 -1 -1
at 572 cm (17.48 p) and baseline from 610 cm to 520 cm (16.4 p
to 19.2 p).
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent dibromochloropropanc as
follows:
-------
Dibromochloropropane EPA-1
_ _(_abs_._ _sample_)_(cgnc. std in mg/ml) (% purity stdl
(abs. std)(cone, sample in
(A concentration of 1 mg dibromochloropropane/ml carbon
distilfide chives an absorbance of apnrox. 0.007 in a 0.2 mm
cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
-------
November 1975
Dibromochloropropane EPA-2
(Tentative)
Determination of Dibromochloropropane
by Gas-Liquid Chromatography (TCD)
Dibromochloropropane is the trivial name for 1, 2-dibromo-3-chloro-
propane, a registered soil fumigant and nematocide having the chemical
structure:
H
H
H
•C C
H
C
Br Br Cl
H
Molecular formula: C.H Br-Cl
Molecular weight: 236.3
Boiling point: 196°C
Physical state, color, and odor: amber to dark brown dense liquid
with a mildly pungent odor
Solubility: 1000 ppm in water; miscible with aliphatic and aromatic
solvents
Stability: stable to hydrolysis in neutral or acid media; hydrolyzed
by alkali to 2-bromoallyl alcohol; corrodes aluminum,
magnesium, and tin alloys
Other names: Fumazone (Dow Chemical Co.), Nemagon (Shell Development Co.),
DBCP, Nemafume, BBC 12, OS 1897
-------
2 Dibromochloropropane EPA-2
(Tentative)
Reagents:
1. Dibromochloropropane standard of known % purity
2. Chloroform, pesticide or spectro grade
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 5' x 1/4" O.D. glass column packed with 20% SE-30 on
Chromosorb W AW DMCS (or equivalent column)
3. Precision liquid syringe: 50 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for TCP-
Column temperature• 140°C
Injection temperature: 175°C
Detector temperature: 175°C
Carrier gas: Helium
Flow rate: 40 ml/min
Operating conditions for filament current, column temperature, or
gas flow should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure:
Preparation of Standard:
Weifjh 0.20 gram dibromochloropropane standard into a 10 ml
volumetric flask and make to volume with chloroform. (final cone
20 mg/ml)
-------
Dibromochloropropane E
(Tentative)
Preparation of Sample:
For technical material and liquid formulations, weigh a portion
of sample equivalent to 0.20 gram dibronochloropropane into a 10 ml
volumetric flask, make to volume with chloroform, and mix thoroughly.
(final cone 20 mg dibromochloropropane/ml)
For dry formulations, weigh a portion of sample equivalent to
1.0 gram dibromochloropropane into a 125 ml screw-cap flask, add by
pipette 50 ml chloroform, and shake for one hour. Allow to settle;
filter or centrifuge if necessary, taking precautions to prevent
evaporation. (final cone 20 mg dibromochloropropane/ml)
Determination:
Using a precision liquid svringe, alternately inject three
30-40 pi portions each of standard and sample solutions. Measure
the peak height or peak area for each peak and calculate the
average for both standard and sample.
Adjustments in attenuation or amount injected mav have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the percent
dibromochloropropane as follows:
ht_. or area sample) (wt. std injected) (% purity of std)
(pk. ht. or area standard) (wt . sample injected)
Method submitted by Eva Santos, EPA Region IX, San Francisco, California
-------
January 1976 Dibutyl Succinate EPA-1
Determination of Dibutyl Succinate
by Saponification and Titration
Dibutyl succinate is a registered insect repellent with the following
chemical structure:
0
CH2 C—0—CH2 CH2— CH2 CH3
0
II
CH2 C—0—CH2 CH2 — CH2—CH3
Molecular formula: C,0H^/10.
12 22 4
Molecular weight: 230.3
Melting/boiling point: m.p.-29°C; b.p. 108°C at A mm Hg
Physical state and color: colorless liquid
Solubility: practically insoluble in water but miscible with most
organic solvents including petroleum oils
Stability: non-corrosive; hydrolyzed by alkalis
Other names: Tabutrex, renamed Tabatrex (Glen Chemical Co.)
Dibutyl succinate is normally formulated with oleic acid (cis-9-
octadecenoic acid) to prolong activity. Oleic acid has the chemical
structure:
CH3 (CH2)7—CH=CH—(CH2)?—COOH
-------
2 Dibutyl Succinate EPA-1
tfclecular formula: C,_H_.00 Molecular weight: 282.45
lo J4 /
Colorless liquid; solidifies at 4°C to crystalline mass;
soluble in alcohol, benzene, chloroform, ether, fixed & volatile oils
Principle of the Method:
The total acidity in the sample is determined by titration with
standard acid after saponification of the dibutyl succinate with an
excess of standard alkali. Any free acid is determined by direct titra-
tion with standard alkali. The difference (as milliequivalents) is equal
to the dibutyl succinate. The free acid is calculated as oleic acid.
Reagents:
1. Sodium (or potassium) hydroxide, 0.5N standard solution
2. Hydrochloric acid, 0.5N standard solution
3. Ethanolic potassium hydroxide, 0.5N standard solution in
ethanol
4. Ethyl alcohol, neutralized to phenolphthalein
5. Phenolphthalein indicator solution
Equipment:
1. Alkali-resistant Erlenmeyer flask, 250-300 ml standard taper
joint
2. Refluxing apparatus
3. Titrating apparatus
4. Usual laboratory glassware
-------
3 Dibutyl Succinate EPA-1
Procedure:
Total acidity after hydrolysis:
Weigh a portion of sample equivalent to 0.3-0.5 gram dibutyl
succinate into a 250-300 ml alkali-resistant Erlenmeyer standard
taper flask and add 50,0 ml 0.5N alcoholic potassium hydroxide
solution. To a second identical flask, add 50.0 ml of the same
solution for a blank. Connect each flask to a reflux condenser
and reflux 2 hours. Cool, add several drops of phenolphthalein
indicator solution, and titrate each flask with 0.5N standard
hydrochloric acid. The difference between the two titrations
represents the total acidity after hydrolysis.
Free acidity before hydrolysis:
Weigh a portion of sample equivalent to 0.3-0.5 gram dibutyl
succinate into a 250-300 ml Erlenmeyer flask. Add 50 ml neutra-
lized alcohol, several drops of phenolphthalein solution, and
titrate with 0.5N standard sodium (or potassium) hydroxide. The
titration represents any free acid and is calculated as oleic acid.
Calculations:
A = milliequivalents of total acid after hydrolvsis
A = (ml HC1 for Blank-mi HC1 for Sample)(N HC1)
B = milliequivalents of free acid before hydrolysis
B = (ml NaOH)(N NaOH)
v n-u . i 4 » (A - B)(0.11515)(100)
% Dibutyl succinate = ; r^r
(grams sample)
(milliequivalent weight of dibutyl succinate = 0.11515)
% Oleic acid = IB) (0.282^) (100)
(grams sample)
(milliequivalent weight of oleic acid = 0.28245)
-------
October 1975
Dichlobenil EPA-1
Determination of Dichlobenil
by Infrared Spectroscopy
Dichlobenil is the accepted common name for 2,6-dichlorobenzo-
nitrile, a registered herbicide having the chemical structure:
Molecular formula: C H Cl N
Molecular weight: 171.9
Melting point:
145-146°C for pure compound; the technical product
is about 95% pure and has a m.p. 139 to 146°C
Physical state, color, and odor: white crystalline solid with an
aromatic odor; technical is gray-white to
yellow-brown
Solubility: very slightly soluble in water (18 ppm at 20°C); slightly
soluble in most organic solvents
Stability: stable to heat and acids but is hydrolyzed by alkalis to
2,6-dichlorobenzamide; non-corrosive; compatible with
other herbicides
Other names: Casoron (N.V. Phillips Duphar), 2,6-DBN, H133
Reagents;
1. Dichlobenil standard of known % purity
2. Carbon disulfide
3. Sodium sulfate, anhydrous, granular
-------
2 Dichlobenil EPA-1
Equipment;
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.2 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, put in 1-2 grams anhydrous sodium sulfate, and
seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.1 gram dichlobenil standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml carbon disulfide
by pipette, close tightly, and shake to dissolve. Add a small
amount of anhydrous sodium sulfate to insure dryness. (final
cone 10 mg/ml)
Preparation of Sample:
Weigh an amount of sample equivalent to 0.5 gram dichlobenil
into a glass-stoppered flask or screw-cap tube. Add 50 ml carbon
disulfide by pipette and 1-2 grams anhydrous sodium sulfate.
Close tightly and shake for one hour. Allow to settle; centrifuge
or filter if necessary, taking precautions to prevent evaporation.
(final cone 10 dichlobenil/ml)
-------
Dichlobenil EPA-1
Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings, scan both the standard
and sample from 835 cm~ to 725 cm (12.0 u to 13.8 ji).
Determine the absorbance of standard and sample using the
peak at 806.5 cm (12.4 ;a) and baseline from 819.7 cm to
787.4 cm"1 (12.2 p. to 12.7 ji) .
Calculation;
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent dichlobenil as
follows:
y - (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg dichlobenil/ml carbon disulfide gives
an absorbance of approx. 0.017 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services.
Eva Santos, EPA Region IX, San Francisco, California, submitted a
similar method using:
solvent: chloroform
cone: 10 mg/ml
scan range: 870 cm" to 720 cm" (11.5 p to 13.9 ji)
analytical peak: 780.0 cm (12.82 ji)
baseline: 819.7 cnT1 to 740.7 cm"1 (12.2 p to 13.5 ;i)
Any criticism, comparison, or suggestions as to preference,
accuracy, or precision of using either of these peaks or using CS
or CHC1- will be appreciated.
-------
August 1975
Dichlone EPA-1
Determination of Dichlone
by Infrared Spectroscopy
Dichlone is the official common name for 2,3-dichloro-l,4-
naphthoquinone, a registered fungicide having the chemical
structure:
Cl
Molecular formula: C...H.C1-0.
10 4 22
Molecular weight: 227.1
Melting point: 193°C (slowly sublimes above 32°C)
Physical state and color: yellow crystals or leaflets
Solubility: practically insoluble in water (0.1 ppm at 25°C);
moderately soluble (about 4%) in xylene and 0-dichloro-
benzene; slightly soluble in acetone, benzene, ether,
dioxane.
Stability: stable to light and acids but hydrolyzed by alkali;
incompatible with petroleum oils, DNOC, and lime sulfur;
non-corrosive
Other names: Phygon and Uniroyal (Uniroyal Inc.), USR 604
-------
2 Dichlone EPA-1
Reagents:
1. Dichlone standard of known % purity
2. Acetone, pesticide or spectro grade
3. Chloroform, pesticide or spectro grade
4. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm NaCl or KBr cells
A
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Water bath 40°C
*
5. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Note! This method is applicable in presence of sulfur but
not in presence of Ferbam.
Preparation of Standard:
Weigh 0.075 gram dichlone into a glass-stoppered flask or
screw-cap bottle, add 50 ml chloroform by pipette, and shake to
dissolve. Add a small amount of anhydrous sodium sulfate to
insure dryness. (final cone. 1.5 mg/ml)
-------
3 Dichlone EPA-1
Preparation of Sample:
For 50% wettable powders or other high % formulations, weigh
a portion of sample equivalent to 0.075 gram dichlone into a
glass-stoppered flask or screw-cap bottle; add 50 ml chloroform
by pipette and 1-2 grams anhydrous sodium sulfate. Close tightly
and shake for one hour. Allow to settle; centrifuge or filter
if necessary, taking precaution to prevent evaporation. (final
cone 1.5 mg dichlone/ml)
For 1-4% dusts or other low % formulations, weigh a portion
of sample equivalent to 0.03 gram dichlone into a glass-
stoppered flask or screw-cap bottle; add 50 ml acetone by
pipette and 1-2 grams anhydrous sodium sulfate. Close tightly
and shake for one hour. Allow to settle; centrifuge or filter
if necessary, taking precaution to prevent evaporation. Evap-
orate a 25 ml aliquot over a water bath at 40°C using a gentle
stream of air. Evaporate the last few ml at RT using air only.
Dissolve in about 5 ml chloroform, transfer to a 10 ml volumetric
flask, and make to volume with chloroform. Mix thoroughly and
add a small amount of anhydrous sodium sulfate to insure dryness.
(final cone 1.5 mg dichlone/ml)
Determination:
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1330 cm to
1225 cm"1 (7.52 p to 8.16 u).
Determine the absorbance of standard and sample using the
peak at 1275 cm" (7.84 ji) and basepoint at 1300 cm" (7.69 /i) .
-------
Dichlone EPA-1
Calculation;
From the above absorbances and using the standard and
sample concentrations, calculate the percent dichlone as
follows:
(abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg dichlone/ml chloroform gives
an absorbance of approx. 0.080 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia, 23219.
-------
September 1975 p-Dichlorobenzene EPA-1
(Tentative)
Determination of p-Dichlorobenzene
by Infrared Spectroscopy
p-Dichlorobenzene (or paradichlorobenzene) is the common name
for 1,4-dichlorobenzene, a fumigant having the chemical structure:
Molecular formula: C H.C1
Molecular weight: 147.01
Melting point: 53°C
Boiling point: 173.4°C
Physical state, color, and odor: colorless crystals with a character-
istic penetrating odor
Solubility: about 80 ppm in water at 25°C; slightly soluble in cold
alcohol; readily soluble in organic solvents
Stability: stable; sublimes at ordinary temperatures; non-corrosive
and non-staining
Other names: Paradow, Paracide, PDB, Santochlor
Reagents:
1. p-Dichlorobenzene standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
p-Dichlorobenzene EPA-1
(Tentative)
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.5 mm KBr or NaCl cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.125 gram p-dichlorobenzene standard into a small
glass-stoppered flask or screw-capped bottle. Add 50 ml
chloroform by pipette and shake to dissolve. Add a small
amount of anhydrous sodium sulfate to insure dryness. (final
cone 2.5 rag/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.125 gram p-dichloro-
benzene into a small glass-stoppered flask or screw-cap bottle.
Add 50 ml chloroform by pipette and 1-2 grams anhydrous sodium
sulfate. Close tightly and shake for one hour. Allow to settle;
centrifuge or filter if necessary, taking precaution to prevent
evaporation. (final cone 2.5 mg p-dichlorobenzene/ml)
Determination:
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 870 cm to
740 cm'1 (11.5 u to 13.5 ;»).
Determine the absorbance of standard and sample using the peak
at 816 cm"1 (12.25 p) and baseline from 855 cm"1 to 794 cm'1 (11.7
to 12.6 i).
-------
p-Dichlorobenzene EPA-1
(Tentative)
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent p-dichlorobenzene as
follows:
(abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method contributed by Nancy Frost, EPA Region IX, San Francisco,
California.
-------
October 1975
p-Dichlorobenzene EPA-2
(Tentative)
Determination of p-Dichlorobenzene
by Gas-Liquid Chromatography
(TCD - Internal Standard)
p-Dichlorobenzene (or paradichlorobenzene) is the common name
for 1,4-dichlorobenzene, a fumigant having the chemical structure:
Molecular formula: C,H.C1~
64 2
Molecular weight: 147.01
Melting point: 53°C
Boiling point: 173.4°C
Physical state, color, and odor: colorless crystals with a character-
istic penetrating odor
Solubility: about 80 ppm in water at 25°C; slightly soluble in cold
alcohol; readily soluble in organic solvents
Stability: stable; sublimes at ordinary temperatures; non-corrosive
and non-staining
Other names: Paradow, Paracide, PDB, Santochlor
-------
2 p-Dichlorobenzene EPA-2
(Tentative)
Reagents:
1. p-Dichlorobenzene standard of known % purity
2. DDVP standard of known % purity
3. Benzene, pesticide or spectro grade
A. Internal Standard solution - weigh 1.8 grams DDVP into a 50 ml
volumetric flask; dissolve in and make to volume with benzene.
(cone 36 mg DDVP/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 6' x 1/8" stainless steel column packed with 10%
SE-30 on Diatoport S (or equivalent or suitable
column)
3. Precision liquid syringe: 5 jil
4. Usual laboratory glassware
Operating Conditions for TCD;
Column temperature: 117°C
Injection temperature: 140°C
Detector temperature: 140eC
Filament current: 190 ma
Carrier gas: Helium
Carrier gas pressure: (not stated in method)
Carrier gas flow rate: (not stated in method)
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
p-Dichlorobenzene EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.088 gram p-dichlorobenzene standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 10 ml of the
internal standard solution and shake to dissolve. (final cone
8.8 mg p-dichlorobenzene and 36 mg DDVP/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.088 gram p-dichloro-
benzene into a small glass-stoppered flask or screw-cap bottle.
Add by pipette 10 ml of the internal standard solution. Close
tightly and shake thoroughly to dissolve and extract the p-dichloro-
benzene. For coarse or granular materials, shake mechanically for
10-15 minutes or shake by hand intermittently for 25-30 minutes.
(final cone 8.8 mg p-dichlorobenzene and 36 mg DDVP/ml)
Determination:
Inject 1-2 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within approx. 10 minutes and peak heights of from 1/2
to 3/4 full scale. The elution time of p-dichlorobenzene is
approx. 1.3 minutes and that of DDVP approx. 4.5 minutes.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of p-dichlorobenzene and
DDVP from both the standard-internal standard solution and the
sample-internal standard solution.
-------
p-Dichlorobenzene EPA-2
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
* PDB = p-dichlorobenzene in following calculation formulas
RF = (wt. DDVP)(% purity DDVP)(pk. ht. or area PDB)
(wt. PDB)(% purity PDB)(pk. ht. or area DDVP)
Determine the percent p-dichlorobenzene for each injection of
the sample-internal standard solution as follows and calculate
the average:
= (wt. DDVP)(% purity DDVP)(pk. ht. or area PDB)
(wt. sample) (pk. ht. or area DDVP) (RF) / LJ-/J
Method submitted by Stelios Gerazounis, EPA Region II, New York, N. Y.
-------
October 1975
Dicloran EPA-1
Determination of Dicloran in Dusts and
Wettable Powder by Infrared Spectroscopy
Dicloran is the common name for 2,6-dichloro-4-nitroaniline, a
registered fungicide having the chemical structure:
Molecular formula: C H.C1-N 0
Molecular weight: 207
Melting point: 192 to 194°C
Physical state, color, and odor: odorless, yellow crystalline solid;
the technical product is brownish-yellow and is
at least 90% pure.
Solubility: practically insoluble in water; slightly soluble in
non-polar solvents; moderately soluble in polar solvents,
e.g., acetone, 3.4 g/100 g at 20°C
Stability: stable to hydrolysis and to oxidation; non-corrosive;
non-flammable; compatible with other pesticides
Other names: Allisan (Boots Company Ltd.), Botran (Upjohn Co.), DCNA,
ditranil
Reagents;
1. Dicloran standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Ethyl ether, pesticide or spectro grade
4. Sodium sulfate, anhydrous, granular
-------
2 Dicloran EPA-1
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.5 mm KBr or NaCl cells
2. Mechanical shaker
3. Soxhlet extraction apparatus
4. Centrifuge or filtration apparatus
5. Rotary evaporator
6. Cotton or glass wool
7. Usual laboratory glassware
Procedure:
Preparation of Standard;
Weigh 0.1 gram dicloran standard into a small glass-
stoppered flask or screw-cap bottle, add 25 ml chloroform
by pipette, and shake to dissolve. Add a small amount of
anhydrous sodium sulfate to insure dryness. (final cone 4 mg/ml)
Preparation of Sample;
For high percent formulations (more than 10%), weigh a
portion of sample equivalent to 0.2 gram dicloran into a glass-
stoppered flask or screw-cap bottle. Add 50 ml chloroform by
pipette and 1-2 grams anhydrous sodium sulfate. Close tightly
and shake for one hour. Allow to settle; centrifuge or filter
if necessary, taking precaution to prevent evaporation. (final
cone 4 mg dicloran/ml)
For low percent (less than 10%) formulations, weigh a
portion of sample equivalent to 0.2 gram dicloran into a Soxhlet
extraction thimble, plug with cotton or glass wool, and extract
with ethyl ether for 1-2 hours. Evaporate the ethyl ether
-------
3 Dicloran EPA-1
completely on a rotary evaporator. Dissolve the residue, transfer
to a 50 ml volumetric flask, and make to volume with chloroform.
Add a small amount of anhydrous sodium sulfate to clarify and
dry the solution, (final cone 4 mg dicloran/ml)
De terminat ion:
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1250 cm to
1042 cm'1 (8 p. to 9.6 ji).
Determine the absorbance of standard and sample using the
peak at 1147 cm~ (8.72 ji) and baseline from 1183 cm" to 1100 cm"
(8.45 u to 9.09 u).
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent dicloran as follows:
-/ m (abs. sample) (cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
-------
January 1976
Dinocap EPA-1
Determination of Dinocap
by Total Nitrogen Analysis
Dinocap is a common name for an isomeric mixture of 2,4-dinitro-
6-octylphenyl crotonate (I) and 2,6-dinitro-4-octylphenyl crotonate (II),
octyl being a mixture of 1-methylheptyl, 1-ethylhexyl, and 1-propyl-
pentyl isomers. The chemical structures are:
0
0-C-CH=CH-CH3
CH3-(CH2)S—CH-(CH2)rCH3
(II)
Dinocap is a registered acaricide and fungicide.
Molecular formula: C1QH_.N 0
IB ZH 2. o
Molecular weight: 364
Boiling point: 138 to 140°C at 0.05 mm Hg
Physical state and color: dark brown liquid
Solubility: practically insoluble in water, soluble in most organic
solvents
Stability: compatible with most other fungicides and insecticides but
should not be used with oil-base sprays or with lime-sulfur
-------
2 Dinocap EPA-1
Other names: Karathane, Arathane (Rohm & Haas); Isocothan, Mildex
Principle of the Method;
Since the nitrogen is present in the nitro (oxidized) form, it must
be converted to the amino (reduced) form before being determined by the
regular Kjeldahl procedure. This is done by reacting the sample with
salicylic acid and concentrated sulfuric acid to form nitro salicylic
acid. The addition of a reducing agent such as zinc then reduces the
nitro group to an amine group, forming amino salicylic acid. This
compound is digested with boiling concentrated sulfuric acid in the
presence of an oxidizing catalyst and forms ammonium sulfate from the
amino-nitrogen. The solution is then made strongly alkaline and the
released ammonia is distilled and absorbed in standard acid.
Reagents;
1. Concentrated sulfuric acid, reagent grade
2. Salicylic acid, reagent grade
3. Zinc dust, reagent grade
4. Mercuric oxide, red, reagent grade
(Commercial packages called "Kel-pacs" are available containing
various oxidizing catalysts and various amounts of potassium
sulfate in small oxidizable plastic packets. One packet can be
dropped into the flask, saving the weighing and transfer of the
HgO and K2S04.)
5. Potassium sulfate, reagent grade (see above)
6. Sodium or potassium sulfide, reagent grade
7. Granulated zinc, reagent grade
-------
3 Dinocap EPA-1
8. Kjeldahl sodium hydroxide solution (450 grams NaOH free from
nitrates in one liter of water)
9. Phenolphthalein indicator solution
10. Sulfuric acid, 0.1N standard solution
(An alternate procedure is to use 50 ml of a saturated boric
acid solution that simply holds the ammonia which is titrated
with standard acid. The procedure eliminates the need for
standard alkali solution.)
11. Sodium hydroxide, 0.1N standard solution (see above)
12. Mixed methyl red indicator solution - dissolve 1.25 grams
methyl red and 0.825 gram methylene blue in one liter of 90%
ethyl alcohol. The color change is from purple in acid to
green in basic solution.
Equipment;
1. 800 ml Kjeldahl flask
2. Kjeldahl digestion and distillation apparatus
(Although a commercial Kjeldahl digestion and distillation
apparatus is convenient, it is not essential. The digestion
may be conducted over a flame in a hood while the distillation
may utilize only a trap and condenser.)
3. Titration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Sample:
Most well mixed or homogeneous samples may be used directly for
analysis; however, low percent formulations such as a 1% dust, or
formulations containing any nitrogenous plant material should be
extracted on a Soxhlet or by shaking with chloroform.
-------
Dinocap EPA-1
Reduction of NO Group:
Weigh a portion of sample equivalent to 0.3-0.A gram technical
dinocap into an 800 ml Kjeldahl flask. Add 35 ml concentrated
sulfuric acid containing 2 grams salicylic acid and allow to stand
at least 30 minutes with frequent shaking.
Add 2 grams zinc dust slowly, shaking the contents of the flask
during the addition. Heat over a low flame until frothing ceases,
then heat until the acid boils briskly for about 5 minutes.
Digestion:
Add 0.7 gram mercuric oxide and 10 grams potassium sulfate (or
one Kel-pac) and continue boiling until the liquid in the flask has
been colorless for one hour. If the contents of the flask tend to
become solid before this point is reached, add 10 ml more of sul-
furic acid. To avoid decomposition of ammonium sulfate and sub-
sequent loss of ammonia, do not allow the flame to reach any part
of the flask not in contact with liquid. The flask may be lifted
from the digestion rack and the acid swirled around the inside of
the flask to wash undigested particles back into the acid. When
digestion is complete, cool and add 200-300 ml water, making sure
that the digestion mixture is completely dissolved.
Distillation;
Measure 50.00 ml of standard 0.1N sulfuric acid into a 500 ml
Erlenmeyer wide-mouth flask, add several drops of mixed methyl red
indicator solution, and place under the condenser of the distilling
apparatus, making sure that the condenser tube extends beneath the
surface of the acid in the flask. A glass tube attached by inert
tubing to the condenser outlet tube is very convenient when later
removing the receiving flask. If the indicator changes from acidic
(purple) to basic (green), the determination must be repeated using
less sample or more acid in the receiving flask.
-------
5 Dinocap EPA-1
Add 25 ml sodium or potassium sulfide solution and mix thor-
oughly; then add several pieces of granulated zinc.
(When using mercury as a catalyst, it must be
precipitated with K or Na sulfide before the
distillation process since it forms a complex
substance with ammonia which is not readily
decomposed by alkali.)
(Zinc in an alkaline solution slowly reacts to
form a zincate and hydrogen: Zn + 2NaOH > Na2ZnO_ + H T
This slow evolution of hydrogen keeps the solution
stirred, thereby preventing superheating.
Pour about 110 ml of the Kjeldahl sodium hydroxide solution (or
if extra acid was added, use 25 ml more alkali for each 10 ml acid
added) slowly down the inclined neck of the flask so that it layers
under the acid solution without mixing. A few drops of phenol-
phthalein may be added to be sure sufficient alkali is added to
neutralize all the acid, remembering that a considerable excess of
alkali will destroy the pink color.
Connect the flask to the condenser by means of a Kjeldahl
connecting bulb, ignite the burner, and quickly mix the contents of
the flask thoroughly with a rotary motion. It is advisable to begin
the distillation with a small flame until the solution begins to
boil; then increase the heat until the solution boils briskly. Dis-
till 150-200 ml of the liquid (the first 150 ml usually contains
all of the ammonia) into the receiving flask. Move the flask so
that the tip of the delivery tube is above the level of the liquid
and distill another 10 ml or so to wash the inside of the tube.
Shut off heat, wash the outside of the delivery tube, and remove
flask from apparatus.
-------
6 Dinocap EPA-1
Titration and Calculation:
Titrate the excess standard acid with standard 0.1N sodium
hydroxide using mixed methyl red indicator. Reagents for this
determination should be acid-free or a reagent blank should be
run. Calculate the percent nitrogen as follows:
Using a blank;
% = (ml NaOH for blank - ml NaOH for sample)(N of NaOH)(.01401)(100)
(grams of sample)
Not using a blank;
[(ml H SO,)(N of H S0.)-(ml NaOH)(N of NaOH)](.01401)(100)
°t — t H
(grams of sample)
The % dinocap is found by dividing the percent nitrogen by the
percent nitrogen in dinocap.
» ,. % nitrogen in sample
/<, dinocap = 7—7 *—
b. b
Technical dinocap contains from 6.6% to 7.2% nitrogen according
to information received from the Rohm and Haas Company, March 1974.
-------
January 1976
Dinocap EPA-2
(Tentative)
Determination of Dinocap
by Infrared Spectroscopy
Dinocap is a common name for an isomeric mixture of 2,4-dinitro-
6-octylphenyl crotonate (I) and 2,6-dinitro-4-octylphenyl crotonate
(II), octyl being a mixture of 1-methylheptyl, 1-ethylhexyl, and
1-propylpentyl isomers. The chemical structures are:
0
0-C-CH=CH-CH3
0
II
0-C-CH=CH-CH3
(H)
Dinocap is a registered acaricide and fungicide.
Molecular formula: C,QH-.N-0,
18 24 2 6
Molecular weight: 364
Boiling point: 138 to 140°C at 0.05 mm Hg
Physical state and color: dark brown liquid
Solubility: practically insoluble in water, soluble in most organic
solvents
Stability: compatible with most other fungicides and insecticides but
should not be used with oil-base sprays or with lime-sulfur
Other names: Karathane, Arathane (Rohm & Haas); Isocothan, Mildex
-------
2 Dinocap EPA-2
(Tentative)
Reagents;
1. Dinocap standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.1 ram NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, put in 1-2 grams anhydrous sodium sulfate, and
seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure;
Preparation of Standard:
Weigh 0.1 gram dinocap standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml chloroform by pipette, close
tightly, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 10 mg/ml)
-------
3 Dinocap EPA-2
(Tentative)
Preparation of Sample;
For dusts, granules, and wettable powders, weigh a portion of
sample equivalent to 0.5 gram dinocap into a 125 ml glass-stoppered
or screw-cap Erlenmeyer flask. Add 50 ml chloroform by pipette and
1-2 grams anhydrous sodium sulfate. Close tightly, shake on a
mechanical shaker for 1 hour, allow to settle; filter or centrifuge
if necessary,taking precaution to avoid evaporation, (final cone
10 mg dinocap/ml)
For emulsifiable concentrates and liquid formulations, weigh a
portion of sample equivalent to 0.5 gram dinocap into a 125 ml
glass-stoppered flask or screw-cap bottle. Add 50 ml chloroform by
pipette and sufficient anhydrous sodium sulfate to clarify and dry
the solution. Close tightly, shake a few minutes, add more sodium
sulfate if needed, and shake vigorously on a mechanical shaker for
one hour. Allow to settle; filter or centrifuge if necessary to
get a clear chloroform solution, taking precaution to prevent
evaporation. (final cone 10 mg dinocap/ml)
(There may be interference from the emulsifier in the
sample; if so, another procedure must be used.)
Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings, scan both the standard
and sample from 1430 cm" to 1250 cm" (7.0 p to 8.0 ji).
Determine the absorbance of standard and sample using the peak
at 1340 cm"1 (7.46 p) and baseline from 1385 cm to 1310 cm"
(7.22 u to 7.63 ;i) .
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent dinocap as follows:
-------
Dinocap EPA-2
(Tentative)
(abs. sample)(cone. std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg dinocap/ml chloroform gives an
absorbance of approx. 0.029 in a 0.1 mm cell.)
This method was submitted by the Commonwealth of Virginia, Division
of Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
Note! This method has been designated as tentative since it is a
Va. Exp. method and because some of the data has been suggested
by EPA's Beltsville Chemistry Lab. Any comments, criticisms,
suggestions, data, etc. concerning this method will be appreciated,
-------
February 1976
Dinoseb EPA-1
(Tentative)
Determination of Dinoseb in Formulations
by Infrared Spectroscopy
Dinoseb is the accepted common name for 2-sec-butyl-4,6-dinitro-
phenol, a registered herbicide having the chemical structure:
OH
— CH-CH2-CH3
Molecular formula: C nH N,,0
Molecular weight: 240.2
Melting point: see below
Physical state, color, and odor: pure compound - yellow crystals
mp 38 to 42°C; technical compound - orange-brown
liquid of 95 to 98% purity and mp 30 to 40°C;
pungent odor
Solubility: about 50 ppm in water; soluble in petroleum oils and most
organic solvents; forms salts with inorganic and organic
bases, some of which are water-soluble
Stability: corrosive to mild steel in the presence of water; combusti-
ble, flash point 177°C
Other names: Premerge (Dow), dinosebe (France), Bansanite, Chemox,
Gebutox, DNBP, Dinitro, DN289, Kiloseb, Nitropone, Sinox
-------
2 Dinoseb EPA-1
(Tentative)
Reagents:
1. Dinoseb standard of known % purity
2. Carbon disulfide, ACS grade (or better)
3. Sulfuric acid, concentrated, ACS
4. Sodium hydroxide, 1% aqueous solution
5. Hydrochloric acid, concentrated, ACS
6. Ethyl ether, ACS (or better)
7. Sodium sulfate, anhydrous granular, ACS
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.1 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Water bath, 40°C, and a stream of dry air
*
5. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, put in 1-2 grams anhydrous sodium sulfate, and
seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50 RPM
on a standard Patterson-Kelley twin shell blender that has
been modified by replacing the blending shell with a box to
hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.1 gram dinoseb standard into a small glass-stoppered
flask or screw-cap tube, add 10 ml carbon disulfide by pipette,
-------
Dinoseb EPA-1
(Tentative)
close tightly, and shake to dissolve. Add a small amount of
anhydrous sodium sulfate to insure dryness. (final cone 10 mg/ml)
Preparation of Sample:
For oil solutions and emulsifiable concentrates, weigh a
portion of sample equivalent to 0.5 gram dinoseb into a small
glass-stoppered flask or screw-cap tube; add a few drops of con-
centrated sulfuric acid so that the sample is definitely acidic.
Add 50 ml carbon disulfide by pipette, 1 gram anhydrous sodium
sulfate, and shake on a mechanical shaker for several hours.
Allow to settle; centrifuge or filter if necessary to get a
clear solution. (cone 10 mg dinoseb/ml)
For liquid (water) formulations, weigh a portion of sample
equivalent to 0.5 gram dinoseb (free phenol) into a small glass-
stoppered flask or screw-cap tube, add by pipette 50 ml of 1%
sodium hydroxide solution, and shake for one hour. Transfer a
25 ml aliquot (filter before aliquoting if necessary) to a 125 ml
separatory funnel, dilute to 50 ml, and acidify with hydrochloric
acid, adding several ml in excess. Extract with three 10 ml portions
*
of carbon disulfide , filtering each through a small cotton plug
(moistened with carbon disulfide) into a 100 ml beaker. Evaporate
to less than 25 ml and transfer quantitatively into a 25 ml volu-
metric flask. Make to volume and add a little anhydrous sodium
sulfate to insure dryness. (final cone 10 mg dinoseb/ml)
* Ethyl ether is the recommended extraction
solvent; however, it must be evaporated completely.
The use of carbon disulfide has been suggested as
an alternative procedure and if satisfactory is
more convenient.
-------
Dinoseb EPA-1
(Tentative)
Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings, scan both the standard
and sample from 1430 cm to 1280 cm (7.0 u to 7.8 u).
Determine the absorbance of standard and sample using the
peak at 1340 cm (7.46 ji) and baseline 1390 cm to 1290 cm
(7.19 ji to 7.75 ;i).
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent dinoseb as follows:
„ _ (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
This method has been given a tentative designation because of the al-
ternative use of carbon disulfide instead of ethyl ether as extraction
solvent.
-------
February 1976
Dinoseb EPA-2
(Tentative)
Determination of Dinoseb in Formulations
by Gas-Liquid Chromatography - TCD
Dinoseb is the accepted common name for 2-sec-butyl-4,6-dinitro-
phenol, a registered herbicide having the chemical structure:
OH
N02—1
CH—CH2-CH3
Molecular formula: Clf)H N 0
Molecular weight: 240.2
Melting point: see below
Physical state, color, and odor: pure compound - yellow crystals
mp 38 to 42°C; technical compound - orange-brown
liquid of 95 to 98% purity and mp 30 to 40°C;
pungent odor
Solubility: about 50 ppm in water; soluble in petroleum oils and most
organic solvents; forms salts with inorganic and organic
bases, some of which are water-soluble
Stability: corrosive to mild steel in the presence of water; combusti-
ble, flash point 177°C
Other names: Premerge (Dow), dinosebe (France), Bansanite, Chemox,
Gebutox, DNBP, Dinitro, DN289, Kiloseb, Nitropone, Sinox
-------
2 Dinoseb EPA-2
(Tentative)
Reagents:
1. Dinoseb standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sulfuric acid, concentrated, ACS
4. Sodium hydroxide, 1% aqueous solution
5. Hydrochloric acid, concentrated, ACS
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 5' x 1/4" glass column packed with 20% SE-30 on
Chromosorb W, AW, DMCS (or equivalent column)
3. Precision liquid syringe: 50 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for TCD;
Column temperature: 220°C
Injection temperature: 250°C
Detector temperature: 250°C
Carrier gas: Helium
Flow rate: adjusted
Operating conditions for filament current, column temperature, or
gas flow should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Dinoseb EPA-2
(Tentative)
Procedure :
Preparation of Standard:
Weigh 0.15 gram dinoseb standard into a 10 ml volumetric
flask; dissolve and make to volume with chloroform. (final cone
15 rag/ml)
Preparation of Sample;
For oil solutions and emulsifiable concentrates, weigh a
portion of sample equivalent to 0.75 gram dinoseb into a small
glass-stoppered flask or screw-cap tube; add a few drops of con-
centrated sulfuric acid so that the sample is definitely acidic.
Add 50 ml chloroform by pipette, and shake on a mechanical shaker
for several hours. Allow to settle; centrifuge or filter if
necessary to get a clear solution. (cone 15 mg dinoseb/ml)
For liquid (water) formulations, weigh a portion of sample
equivalent to 0.75 gram dinoseb (free phenol) into a small glass-
stoppered flask or screw-cap tube, add by pipette 50 ml of 1%
sodium hydroxide solution, and shake for one hour. Transfer a
25 ml aliquot (filter before aliquoting if necessary) to a 125 ml
separatory funnel, dilute to 50 ml, and acidify with hydrochloric
acid, adding several ml in excess. Extract with three 10 ml portions
of chloroform, filtering each through a small cotton plug (moistened
with chloroform) into a 100 ml beaker. Evaporate to less than 25 ml,
transfer quantitatively into a 25 ml volumetric flask, and make to
volume. (final cone 15 mg dinoseb/ml)
Determination;
Using a precision liquid syringe, alternately inject three
30-40 ul portions each of standard and sample solutions. Measure
the peak height or peak area for each peak and calculate the average
for both standard and sample.
Adjustments in attenuation or amount injected may have to be made
to give convenient size peaks.
-------
Dinoseb EPA-2
(Tentative)
Calculation:
From the average peak height or peak area calculate the percent
butylate as follows:
7 - (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
This method is based on a GLC method submitted by Eva Santos, EPA
Region IX, San Francisco, California. The sample preparation is
basically like that of Dinoseb EPA-1 IR method. Any suggestions,
data, criticism, or comments about this method are most welcome.
-------
Diphacinone EPA-1
•November
Determination of Diphacinone in Baits
by Ultraviolet Spectroscopy
Diphacinone is the accepted common name for 2-(diphenylacetyl)-
1,3-indandione, a registered rodenticide having the chemical structure:
^
w/
0
11
\H
,-/
;v_
II
0
Molecular formula: C H,,0
23 16 3
Molecular weight: 340.4
Melting point: 145°C
Physical state, color, and odor: yellow, odorless crystals
Solubility: slightly soluble in water and benzene; soluble in acetone
and acetic acid. Forms a sodium salt which is sparingly
soluble in water.
Stability: resists hydrolysis; stable toward mild oxidants; non-corrosive
Other names: Diphacin (Velsicol Chem. Corp.), diphacin (Turkey), Ramik,
diphenadione
This method is suitable for products containing about 0.005%
diphacinone. Although the absorption curves for diphacinone and pindone
are similar, in the absence of strong interference, diphacinone can be
identified by a maximum at 286 nm and pindone by a maximum at 283 nm.
-------
Diphaclnone EPA-1
(tentative)
Reagents;
1. Diphacinone standard of known % purity
2. Sodium pyrophosphate solutions, 1% and 2% - weigh 5.0
grains for 1% solution and 10.0 grams for 2% solution
into a 500 ml volumetric flask; dissolve in and make
to volume with distilled water (heating on a steam bath
may be required for complete solution).
3. Ethyl ether, pesticide grade
4. Hexane, pesticide grade
5. Ethyl ether-hexane mixture - extract 200 ml hexane with
three 20 ml portions of 1% sodium pyrophosphate solution.
Prepare mixture by adding 20 ml ethyl ether to each 80 ml
extracted hexane.
6. Hydrochloric acid, 50% solution - add 50 ml hydrochloric
acid (specific gravity 1.19) to 50 ml distilled water.
7. Methanol, pesticide grade
8. Acidification Solution - mix equal volumes of methanol
and 50% hydrochloric acid.
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm silica cells
2. Micro-mill or any suitable device for grinding or pulver-
izing sample
3. Bottles with teflon-lined or polyethylene screw caps in
2 oz, 4 oz, and 8 oz sizes
4. Mechanical shaker (wrist action preferred)
-------
Diphacinone EPA-1
(tentative)
5. Centrifuge for bottles and 15-20 ml tubes
6. Syringe, 5 ml capacity with 4-inch needle
7. Usual laboratory glassware
Procedure:
Preparation of Standard;
Prepare a stock standard solution by weighing 0.04 gram
diphacinone into a 100 ml volumetric flask; dissolve in and
make to volume with 1% sodium pyrophosphate solution; mix
well.
For a working standard solution, pipette 1 ml of the
stock standard solution into a 100 ml volumetric flask and
make to volume with 1% sodium pyrophosphate solution; mix
well, (final cone 4 ug diphacinone/ml)
Pipette 25 ml of this working solution into a 4 oz
screw-cap bottle and add 10 ml acidification solution.
By pipette, add 50 ml ether-hexane solution and close
tightly. Proceed as under Determination.
Preparation of Sample;
Weigh a portion of well-ground and mixed sample equiv-
alent to 0.2 mg diphacinone into an 8 oz screw-cap bottle.
(Sufficient sample should be weighed to yield 4 ug/ml in the
final test solution. This is equivalent to 4 grams of 0.005%
product or 0.8 gram of 0.025% product.)
Add 20 ml acidification solution; swirl and mix thoroughly
for 2-3 minutes. Pipette 100 ml of the ether-hexane solution
over the acidified sample and close tightly. Proceed as under
Determination.
-------
Diphacinone EPA-1
(tentative)
Determination:
Place standard and sample on a mechanical shaker (wrist
action preferred) and shake vigorously for one hour. Allow
to settle; transfer a 30 ml aliquot by pipette into a 2 oz
screw-cap bottle. Add by pipette 15 ml 2% sodium pyro-
phosphate solution, close tightly, and shake on shaker for
three minutes. Transfer to a 125 ml separatory funnel and
separate the aqueous (bottom) layer into a 15-20 ml centri-
fuge tube. Centrifuge unstoppered (approx. 3400 RPM) for
about 15 minutes, checking intermittently. Solution must be
clear.
Sample solutions will have a narrow suspended emulsion
layer. This layer may be drawn off using an aspirator fitted
with a glass tube drawn into a fine tip; or, the clear solu-
tion below may be drawn into a 5 ml syringe through the
emulsion layer with a four-inch needle.
UV Determination;
With the UV spectrophotometer at the optimum quantitative
settings for the particular instrument being used, balance the
pen at 0 and 100% transmission at 286 nm with 1% sodium pyro-
phosphate in each cell. Scan the standard and sample solutions
from 360 nm to 200 nm with 1% sodium pyrophosphate solution in
the reference cell. (Distilled water may be used as reference
if desired.)
Calculations;
Measure the absorbance of standard and sample at 286 nm
and calculate the percent diphacinone as follows:
% „ (abs. sample) (cone, standard injjg/ml)(% purity standard)
(abs. standard)(cone, sample in jig/ml)
-------
Diphacinone EPA-1
(tentative)
This method is basically method AM 0556, Velsicol, Analytical
Research Division, Chicago, Illinois 60611, and is used with
their permission.
This method has been used successfully by EPA's New York
and Beltsville Chemical Laboratories. A fev changes in volume
of aliquots were made for more convenience, and the basic format
was changed to conform with the standard format of methods in
this manual.
Some commercial products may present problems with this
method because of Interfering substances, but for most products
this method has been found satisfactory.
Comments, criticisms, suggestions, etc. will be appreciated.
-------
November 1975 Disulfoton EPA-1
(Tentative)
Determination of Disulfoton
by Infrared Spectroscopy
Disulfoton is the common name for 0,0-diethyl S-[2-(ethylthio)
ethyl] phosphorodithioate, a registered insecticide and acaricide
having the chemical structure:
CH3-CH2-C> S
J \ II
J>P—S-CH2-CH2-S-CH2-CH3
CH3-CH2-CT
Molecular formula: C-H.-0_PS_
o iy i j
Molecular weight: 274.2
Boiling point: 62°C at 0.01 mm Hg
Physical state, color, and odor: colorless, oily liquid with a
characteristic odor of sulfur compounds; the
technical product is a dark yellowish oil.
Solubility: 25 ppm in water at RT; readily soluble in most organic
liquids
Stability: relatively stable to hydrolysis below pH 8.0
Other names: Disyston (Di-Syston in US), Dithio-systox, S-276, Bayer 19639,
(Bayer AG); thiodemeton; M-74 (USSR); Frumin AL; Solvirex
-------
2 Disulfoton EPA-1
(Tentative)
Reagents:
1. Disulfoton standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Acetone, pesticide or spectro grade (dried over sodium sulfate)
4. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.5 mm KBr cells (NaCl useful transmission up to 16 u)
2. Mechanical shaker
3. Rotary evaporator
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.125 gram disulfoton standard into a 25 ml volumetric
flask, dissolve in, and make to volume with carbon disulfide. Add
a small amount of granular anhydrous sodium sulfate and shake.
(final cone 5 mg/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.5 gram disulfoton
into a glass-stoppered or screw-capped 250 ml Erlenmeyer flask.
Add, by pipette, 100 ml of mixed solvent (9+1 carbon disulfide +
dry acetone), and shake on a mechanical shaker for one hour. (Be
careful to avoid any loss of solvent around ground glass joint or
screw cap.) Allow to settle; centrifuge or filter if necessary,
taking precaution to prevent evaporation. Pipette 25 ml into a
standard taper 125 ml Erlenmeyer flask and evaporate on a rotary
-------
Disulfoton EPA-1
(Tentative)
evaporator to just dryness. Add 5 ml carbon disulfide and again
evaporate to dryness. Dissolve in, quantitatively transfer to a
25 ml volumetric flask, and make to volume with carbon disulfide.
Add a small amount of granular anhydrous sodium sulfate and shake.
(final cone 5 mg disulfoton/ml)
IR Determination:
With carbon disulfide in the reference cell and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan both the standard and sample solutions
from 740 cm" to 590 cm" (13.5 ji to 17.0 ^i) . For a qualitative
comparison,run a full scan.
Calculation:
Measure the absorbance of standard and sample at 667 cm
(15.0 u) using a baseline from 730 cm" to 633 cm" (13.7 p to
15.8 ji).
„. _ . ., - (abs. sample) (cone, std in mg/ml) (% purity std)
/<, DlSUilOtOn = -;—. rt-; ; •J~.—rr ' *
(abs. std)(cone, sample in mg/ml)
Method submitted by Dean Hill, EPA Region IX, San Francisco, Calif.
-------
December 1975 Disulfoton EPA-2
(Tentative.)
Determination of Disulfoton by
Gas-Liquid Chromatography
(FID - Internal Standard)
Disulfoton is the .common name for 0,0-diethyl S-[2-(ethylthio)
ethyl] phosphorodithioate, a registered insecticide and acaricide
having the chemical structure:
s
II
P-S-CH2-CH2—S-CH2-CH3
CH3-CH2-0
Molecular formula: C_H,_00PS0
o iy i j
Molecular weight: 27A.2
Boiling point: 62°C at 0.01 mm Hg
Physical state, color, and odor: colorless, oily liquid with a
characteristic odor of sulfur compounds; the
technical product is a dark yellowish oil.
Solubility: 25 ppm in water at RT; readily soluble in most organic
liquids
Stability: relatively stable to hydrolysis below pH 8.0
Other names: Disyston (Di-Syston in US), Dithio-systox, S-276, Bayer 19639,
(Bayer AG); thiodemeton; M-74 (USSR); Frumin AL; Solvirex
-------
Disulfoton EPA-2
(Tentative)
Reagents:
1. Disulfoton standard of known % purity
2. Alachlor standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.25 gram alachlor standard
into a 100 ml volumetric flask, dissolve in, and make to volume
with acetone, (cone 2.5 mg alachlor/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: A' x 2 mm I.D. glass, packed with 5% SE-30 80/100 mesh
Chromosorb W HP (or equivalent column)
3. Precision liquid syringe: 10 11!
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 190°C
Injection temperature: 240°C
Detector temperature: 240°C
Carrier gas: Nitrogen
Carrier gas flow rate: adjusted for specific GC
Hydrogen flow rate: adjusted for specific GC
Air flow rate: adjusted for specific GC
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Disulfoton EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.05 gram disulfoton standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 20 ml of the internal
standard solution and shake to dissolve. (final cone 2.5 rag
disulfoton and 2.5 rag alachlor/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.05 gram disulfoton
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the disulfoton. For
coarse or granular materials, shake mechanically for 30 minutes or
shake by hand intermittently for one hour. (final cone 2.5 mg
disulfoton and 2.5 mg alachlor/ml)
Determination:
Inject 1-2 pi of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is disulfoton, then alachlor.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of disulfoton and alachlor from
both the standard-internal standard solution and the sample-internal
standard solution.
-------
Disulfoton EPA-2
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(_wt_._ alachlor) (% purity alachlor)(pk. ht._or area disulfoton)
(wt. disulfoton)(% purity disulfoton)(pk. ht. or area alachlor)
Determine the percent disulfoton for each injection of the
sample-internal standard solution as follows and calculate the
average:
7 m (wt. alachlor)(% purity alachlor)(pk. ht. or area disulfotxm)
(wt. sample) (pk. ht. or area alachlor) (RF)
Method submitted by the Commonwealth of Virginia, Division of Consol-
idated Laboratory Services, 1 North 14th Street, Richmond, Virginia 23219,
Note! This method has been designated as tentative since it is a Va.
Exp. method and because some of the data has been suggested by
EPA's Beltsville Chemistry Laboratory. Any comments, criticisms,
suggestions, data, etc. concerning this method will be appreciated,
-------
September 1975
Diuron EPA-1
Determination of Diuron by
Alkaline Hydrolysis and Titration
Diuron is the common name for 3-(3,4-dichlorophenyl)-l,l-dimethyl-
urea, a registered herbicide having the chemical structure:
Molecular formula: C-H 0C12N 0
Molecular weight: 233.1
Melting point: 158 to 159°C
Physical state, color, and odor: Odorless, white, crystalline solid
Solubility: 42 ppm in water at 25°C; slightly soluble in hydrocarbons,
about 5.3% in acetone at 27°C
Stability: decomposes at 180-190°C; non-corrosive; stable at ordinary
temp, to oxidation and moisture, hydrolyzes at higher temp.
and more acid or alkaline conditions
Other names: Karmex (DuPont), Di-on, Diurex, Vonduron, dichlorfenidim
Principle of the Method;
The diuron is hydrolyzed to 3,4-dichloroaniline, carbon dioxide
(as carbonate), and dimethylamine. The dimethylamine is distilled and
titrated. Volatile, moderately strong bases, or substances that
hydrolyze to give them, interfere.
-------
2 Diuron EPA-1
Reagents:
1. Potassium hydroxide, 20% solution
2. Hydrochloric acid, 0.1N standard solution
3. Sodium hydroxide, 0.1N standard solution
4. Ethyl alcohol, ACS
5. Glycerol, ACS
Equipment;
1. Distilling apparatus consisting of a 500 ml round-bottom
flask with a thermometer well in the side and a 24/40
standard taper (ST) joint at the top. The flask is
connected to the bottom of a vertical condenser which has
its top connected to the top of a second vertical con-
denser by a horizontal tube with a right angle 24/40 ST
joint on each end. The bottom of the second condenser is
connected by 24/40 ST joint to the top of a delivery tube
which has a narrow plain end extending almost to the
bottom of a receiving beaker.
2. 500 ml size heating mantle with variable transformer control
3. Thermometer to 200°C
4. Potentiometric titrimeter
5. Usual laboratory glassware
Procedure;
Weigh a portion of sample equivalent to 0.4-0.5 gram diuron
into the reaction flask, dissolve in 25 ml ethyl alcohol, and add
100 ml glycerol and 100 ml 20% potassium hydroxide solution. Attach
immediately to the first condenser.
-------
3 Diuron EPA-1
Pipette 50 ml of the 0.1N standard hydrochloric acid into the
receiving beaker. Reflux at a moderate rate for 2-1/2 hours with
water flowing through both condensers. Remove the water from the
first condenser and distill until the temperature at the thermom-
eter well reaches 175°C — usually about 50 minutes. (The
temperature rises rapidly at the end.)
Titration;
Remove the delivery tube and receiving beaker and rinse the
delivery tube into the beaker. Titrate the excess standard acid
with the 0.1N standard sodium hydroxide potentiometrically, using
a glass electrode and a calomel electrode. The inflection point,
which occurs at about pH 7.6, is taken as the endpoint.
With less accuracy, bromthymol blue may be used as an internal
indicator.
Calculation:
Calculate the percentage of diuron as follows:
% „ (ml)(N)(.2331)(100)
(g sample)
where: .2331 is the milliequivalent weight of diuron
(1 ml 0.1N HC1 - 0.02331 g diuron)
This method is based on Lowen and Baker, Anal. Chem. 24, 1475 (1952).
-------
July 1975
Diuron EPA-2
(Tentative)
Determination of Diuron by
High Pressure Liquid Chrotnatography
Diuron is the common name for 3-(3,4-dichlorophenyl)-l,l-dimethyl-
urea, a registered herbicide having the chemical structure:
CH
Molecular formula: C H Cl N 0
Molecular weight: 233.1
Melting point: 158 to 159°C
Physical state, color, and odor: Odorless, white, crystalline solid
Solubility: 42 ppm in water at 25°C; slightly soluble in hydrocarbons,
about 5.3% in acetone at 27 °C
Stability: decomposes at 180-190°C; non-corrosive; stable at ordinary
temp, to oxidation and moisture, hydrolyzes at higher temp.
and more acid or alkaline conditions
Other names: Karmex (DuPont), Di-on, Diurex, Vonduron, dichlorfenidim
Reagents:
1. Diuron standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Methanol, pesticide or spectro grade
-------
2 Diuron EPA-2
(Tentative)
Equipment;
1. High pressure liquid chromatograph
2. High pressure liquid syringe or sample injection loop
3. Liquid chromatographic column 4 mm x 25 cm packed with
Vydac Reverse Phase Hydrocarbon
4. Usual laboratory glassware
Operating conditions for Hewlett-Packard Model 1010B LC;
Mobile phase: 80% methanol + 20% water
Column temperature: ambient
2
Observed column pressure: 30-40 kg/cm (425-570 PSI)
Flow rate: 3 ml/min
Detector: UV at 254 nm
Chart speed: 0.5 in/min
Injection: 10 jil
Conditions may have to be varied by the analyst for other instru-
ments, column variations, sample composition, etc. to obtain optimum
response and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.02 gram diuron standard into a 100 ml volumetric
flask; dissolve and make to volume with chloroform (final
cone 0.2 mg/ml).
-------
3 Diuron EPA-2
(Tentative)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.2 gram diuron
in a 125 ml screw-cap Erlenmeyer flask, add 50 ml chloroform
by pipette, close tightly, and shake for one hour. Let stand
for 30 minutes or until clear (filter or centrifuge if
necessary). Pipette 5 ml of the clear supernatant liquid
into a 100 ml volumetric flask. Make to volume with chloro-
form and mix thoroughly (final cone 0.2 mg diuron/ml).
Determination:
Alternately inject three 10 ul portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent diuron as follows:
•y - (pk. ht. or area sample) (wt. std injected) (% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method developed by Joseph B. Audino, Supervisor, Pesticide Formulation
Laboratory, California Department of Food and Agriculture; and by
Hoshihiko Kawano, Associate Chemist on sabbatical leave from the
University of Hawaii.
-------
November 1975 Diuron EPA-3
(Tentative)
Determination of Diuron
by Ultraviolet Spectroscopy
Diuron is the common name for 3-(3,4-dichlorophenyl)-l,l-dimethyl-
urea, a registered herbicide having the chemical structure:
Molecular formula: C H Cl N 0
Molecular weight: 233.1
Melting point: 158 to 159°C
Physical state, color, and odor: Odorless, white, crystalline solid
Solubility: 42 ppm in water at 25°C; slightly soluble in hydrocarbons,
about 5.3% in acetone at 27°C
Stability: decomposes at 180-190°C; non-corrosive; stable at ordinary
temp, to oxidation and moisture, hydrolyzes at higher temp,
and more acid or alkaline conditions
Other names: Karmex (DuPont), Di-on, Diurex, Vonduron, dichlorfenidim
Reagents:
1. Diuron standard of known % purity
2. Methanol, pesticide or spectro grade
-------
Diuron EPA-3
(Tentative)
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm silica cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard;
Weigh 0.1 gram diuron standard into a 100 ml volumetric flask,
add 100 ml methanol by pipette, and mix thoroughly. Pipette 10 ml
into a second 100 ml volumetric flask, make to volume with methanol,
and mix thoroughly. Pipette 5 ml into a third 100 ml volumetric
flask, make to volume with methanol, and mix thoroughly. (final
cone 5 ug/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.1 gram diuron into a
250 ml glass-stoppered or screw-cap flask, add 100 ml methanol by
pipette, and shake on a mechanical shaker for 30 minutes. Allow
to settle; centrifuge or filter if necessary, taking precautions
to prevent evaporation. Pipette 10 ml into a 100 ml volumetric
flask, make to volume with methanol, and mix thoroughly. Pipette
5 ml of this solution into another 100 ml volumetric flask, make to
volume with methanol, and mix thoroughly. (final cone 5 ug diuron/ml)
UV Determination:
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen at 0 and 100% transmission at 248 nm with
-------
Diuron EPA-3
(Tentative)
methanol in each cell. Scan both the standard and sample from
300 nm to 200 nra with methanol in the reference cell.
Measure the absorbance of standard and sample using the peak
at 248 nm and a basepoint at 280 nm.
The absorbance is linear from 1 to 8 iig/ml.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent diuron as follows:
„ m (abs. sample)(cone, std in jig/ml)(% purity std)
(abs. std) (cone, sample in
Method submitted by Mark Law, EPA, Beltsville Chemistry Laboratory,
Beltsville, Md.
(This method is based on Monuron EPA-2.)
-------
November 1975 Diuron EPA-4
(Tentative)
Determination of Diuron
by Infrared Spectroscopy
Diuron is the common name for 3-(3,4-dichlorophenyl)-l,l-dimethyl-
urea, a registered herbicide having the chemical structure:
Molecular formula: C H Cl N 0
Molecular weight: 233.1
Melting point: 158 to 159°C
Physical state, color, and odor: odorless, white, crystalline solid
Solubility: 42 ppm in water at 25°C; slightly soluble in hydrocarbons,
about 5.3% in acetone at 27°C
Stability: decomposes at 180-190°C; non-corrosive; stable at ordinary
temp, to oxidation and moisture, hydrolyzes at higher temp,
and more acid or alkaline conditions
Other names: Karmex (DuPont), Di-on, Diurex, Vonduron, dichlorfenidim
Reagents:
1. Diuron standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
Diuron EPA-4
(Tentative)
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.5 mm KBr or NaCl cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.06 gram diuron standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml chloroform by pipette, close
tightly, and shake to dissolve. Add a snail amount of anhydrous
sodium sulfate to insure dryness. (final cone 6 mg/ml)
Preparation of Sample:
Weigh an amount of sample equivalent to 0.3 gram diuron into
a glass-stoppered flask or screw-cap bottle. Add 50 ml chloroform
by pipette and 1-2 grams anhydrous sodium sulfate. Close tightly
and shake for one hour. Allow to settle; centrifuge or filter if
necessary, taking precautions to prevent evaporation. (final cone
6 mg diuron/ml)
Determination:
With chloroform in the reference cell, and using the optimum
quantitative analytical settings, scan both the standard and
sample from 1500 cm" to 1300 cm 1 (6.67 ji to 7.7 p).
Determine the absorbance o,f standard and sample using the peak
at 1353 cm~ (7.39 p) and baseline from 1399 cm" to 1316 cm"
(7.15 p to 7.60 p).
The absorbance is linear from 1 to 10 mg/ml.
-------
Diuron EPA-4
(Tentative)
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent diuron as follows:
7 - (abs. sample)^conc.gtd in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method submitted by Mark Law, EPA, Beltsville Chemistry Laboratory,
Beltsville, Md.
(This method is based on Monuron EPA-3.)
-------
-------
November 1975
Endosulfan EPA-1
Determination of Endosulfan
by Alkaline Hydrolysis
Endosulfan is the accepted common name for hexachlorohexahydro-
methano-2,4,3-benzodioxathiepin-3-oxide, a registered pesticide
having the chemical structure:
Cl
s=o
Molecular formula: CnH,Cl,0,S
" D O J
Molecular weight:
Melting point:
406.9
(see below)
Physical state, color, and odor: endosulfan is an odorless white
crystalline solid mixture of two isomers with
mp's of 106°C and 212°C; the technical product
is a brownish crystalline solid, mp 70-100°C,
with a 4:1 ratio of the above isomers. Both
isomers are insecticidally active.
Solubility: practically insoluble in water, but soluble in most
organic solvents
Stability: generally quite stable; decomposition catalyzed by iron;
slowly hydrolyzed by water; sensitive to acid and bases;
compatible with non-alkaline pesticides
-------
2 Endosulfan EPA-1
Other names: Thiodan (Farwerke Hoechst), Beosit, Chlorthiepln, Cyclodan,
Insectophene, Kop-Thiodan, Malix, Thifor, Thiraul, Thionex,
HOE 2671, NIA 5462, FMC 5462
Principle of the Method;
This determination is based on the alkaline hydrolysis of endosulfan
to give sodium sulfite, which is reacted with an excess of acidified
standard iodine solution. The excess iodine solution is titrated with
standard sodium thiosulfate solution and the amount of endosulfan is
calculated from the amount of iodine used by the sodium sulfite.
Reagents:
1. Methanol, ACS
2. n-Hexane, ACS
3. Sodium hydroxide pellets, ACS
4. Sulfuric acid solution, 1+4
5. Sodium hydroxide solution, 1+9
6. Phenolphthalein solution, 1% in alcohol
7. Standard 0.1N iodine solution
8. Standard 0.1N sodium thiosulfate solution
9. Starch solution, 0.2%
Equipment:
1. Iodine titration flasks
2. Refluxing apparatus
3. Mechanical shaker
4. Usual laboratory glassware
-------
3 Endosulfan EPA-1
Procedure:
Preparation of Sample;
For liquid formulations and technical endosulfan, weigh a
portion of sample equivalent to 0.2-0.3 gram of endosulfan into
a standard taper 250 ml Erlenmeyer flask. Add 100 ml methanol
and proceed directly with the hydrolysis.
For dusts and granules, weigh a portion of sample equivalent
to 0.4-0.6 gram of endosulfan into a screw-capped or glass-
stoppered flask, add 100 ml methanol, and shake for 15 minutes.
Pipette 50 ml of clear liquid into a 250 ml standard taper
Erlenmeyer flask, add an additional 50 ml methanol, and proceed
with the hydrolysis.
If the methanol extract is highly colored, repeat the
extraction on another portion of sample using hexane. Pipette
50 ml of the clear extract into a 250 ml standard taper Erlen-
meyer flask, evaporate the hexane to near dryness over a hot
water bath in a hood, cool, and add 100 ml methanol and proceed
with the hydrolysis.
Hydrolysis:
Add 2-3 grams (15 pellets) of sodium hydroxide to the
methanol solution of the sample and reflux gently for two hours.
Wash down the condenser with 20 ml methanol and then with 50 ml
distilled water. Remove from condenser, add a few drops of
phenolphthalein solution, neutralize with 1 + 4 sulfuric acid
solution to just colorless, and restore color with 1+9 sodium
hydroxide to prevent loss of sulfite as S0~.
-------
Endosulfan EPA-1
Titration;
Add 40 ml of standard 0.1N iodine solution to a 500 ml glass-
stoppered iodine flask using a pipette or burette, acidify with
1 ml 1 + 4 sulfuric acid, and while stirring with a magnetic
stirrer, add the sulfite solution slowly. Rinse the flask with
several small portions of distilled water until all the sulfite
is transferred; the washing is complete when there is insufficient
sulfite left in the flask to bleach one drop of 0.1N iodine
solution. The final volume in the flask should be about 225-250 ml.
Titrate the excess iodine with standard 0.1N sodium thiosulfate
solution using 10 ml 0.2% starch solution as indicator and titrating
to the disappearance of the blue color.
Run a blank tltration on 40 ml of standard 0.1N iodine solution
using 175 ml distilled water and 1 ml 1 4- 4 sulfuric acid.
Calculation:
The molecular weight of endosulfan is 406.95 and the milli-
equivalent weight is 0.2305.
Net ml Na2S20 used - ml Na2S2°3 for blank - ml Na^O- for sample
(net ml NaO (N of NaO (.2035) (100)
% endosulfan
(grams of sample)(50/100 see note)
Note: The factor (50/100) is not used for liquid formulations
or technical endosulfan.
-------
November 1975
Endosulfan EPA-2
(Tentative)
Determination of Endosulfan
by Infrared Spectroscopy
Endosulfan is the accepted common name for hexachlorohexahydro-
methano-2,4,3-benzodioxathiepin-3-oxide, a registered pesticide
having the chemical structure:
Cl
Cl
Molecular formula: C H Cl 0 S
Molecular weight:
Melting point:
406.9
(see below)
Physical state, color, and odor: endosulfan is an odorless white
crystalline solid mixture of two isomers with
mp's of 106°C and 212°C; the technical product
is a brownish crystalline solid, mp 70-100°C,
with a 4:1 ratio of the above isomers. Both
isomers are insecticidally active.
Solubility: practically insoluble in water, but soluble in most
organic solvents
Stability: generally quite stable; decomposition catalyzed by iron;
slowly hydrolyzed by water; sensitive to acid and bases;
compatible with non-alkaline pesticides
-------
2 Endosulfan EPA-2
(Tentative)
Other names: Thiodan (Farwerke Hoechst), Beosit, Chlorthiepin, Cyclodan,
Insectophene, Kop-Thiodan, Malix, Thifor, Thimul, Thionex,
HOE 2671, NIA 5462, FMC 5462
Reagents:
1. Endosulfan standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.5 mm NaCl or KBr cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure;
Preparation of Standard:
Weigh 0.06 gram endosulfan into a small glass-stoppered flask
or screw-cap bottle, add 10 ml carbon disulfide by pipette, and
shake to dissolve. Add a small amount of anhydrous sodium sulfate
to insure dryness. (final cone 6 mg/ml)
Preparation of Sample;
For emulsifiable concentrates, weigh a portion of sample
equivalent to 0.06 gram endosulfan into a 10 ml volumetric flask,
make to volume with carbon disulfide, and mix well- Add a small
amount of anhydrous sodium sulfa*te to insure dryness. (final cone
6 mg endosulfan/ml)
-------
3 Endosulfan EPA-2
(Tentative)
For granular formulations, weigh a portion of sample equivalent
to 0.3 gram endosulfan into a. glass-stoppered flask or screw-cap
bottle. Add 50 ml carbon disulfide by pipette and 1-2 grams
anhydrous sulfate. Close tightly and shake for one hour. Allow
to settle; centrifuge or filter if necessary, taking precaution
to prevent evaporation. Add a small amount of anhydrous sodium
sulfate to insure dryness. (final cone 6 mg endosulf an/ml)
Determination;
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan the standard and sample from 1250 cm
to 1110 cm (8 p to 9 jj) .
Determine the absorbance of standard and sample using the
at 1192
(8.3 to 8.5
peak at 1192 cm" (8.39 p) and baseline from 1205 cm~ to 1176 cm~
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent endosulfan as follows:
•y B (abs. sample) (cone, std in mg/ml) (% purity std)
(abs. std) (cone, sample in mg/ml)
Method submitted by M. Frost and M. Conti, EPA Region IX, San Francisco,
California.
-------
November 1975
Endosulfan EPA-3
(Tentative)
Determination of Endosulfan
by Gas-Liquid Chromatography
(TCD - Internal Standard)
Endosulfan is the accepted common name for hexachlorohexahydro
raethano-2,4,3-benzodioxathiepin-3-oxide, a registered pesticide
having the chemical structure:
Cl
CH2—Os
CH2—0'
• S=0
H
Molecular formula: CftH_Cl,0_S
y D D j
Molecular weight: 406.9
Melting point: (see below)
Physical state, color, and odor: endosulfan is an odorless white
crystalline solid mixture of two isomers with
mp's of 106°C and 212°C; the technical product
is a brownish crystalline solid, mp 70-100°C,
with a 4:1 ratio of the above isomers. Both
isomers are insecticidally active.
Solubility: practically insoluble in water, but soluble in most
organic solvents
Stability: generally quite stable; decomposition catalyzed by iron;
slowly hydrolyzed by water; sensitive to acid and bases;
compatible with non-alkaline pesticides
-------
2 Endosulfan EPA-3
(Tentative)
Other names: Thiodan (Farwerke Hoechst), Beosit, Chlorthiepin, Cyclodan,
Insectophene, Kop-Thiodan, Malix, Thifor, Thiraul, Thionex,
HOE 2671, NIA 5462, FMC 5462
Reagents:
1. Endosulfan standard of known % purity
2. Aldrin standard of known HHDN content
3. Acetone, pesticide or spectro grade
(chloroform could also be used)
4. Internal Standard solution - weigh 0.1 gram HHDN into a 50 ml
volumetric flask; dissolve in and make to volume with acetone.
(cone 2 mg HHDN/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 6' x 1/8" ID SS packed with 10% SE-30 on 60/80 mesh
Diatoport S (or equivalent column)
3. Precision liquid syringe - 10 or 25 pi
4. Usual laboratory glassware
Operating Conditions for TCP:
Column temperature: 230°C
Injection temperature: 260°C
Detector temperature: 260°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas pressure: 30-40 psi
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Endosulfan EPA-3
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.08 gram endosulfan standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 20 ml of
the internal standard solution and shake to dissolve, (final
cone A mg endosulfan and 2 rag HHDN/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.08 gram endosulfan
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the endosulfan.
For coarse or granular materials, shake mechanically for 30 minutes
or shake by hand intermittently for one hour. (final cone 4 mg
endosulfan and 2 mg HHDN/ml)
Determination;
Inject 10-15 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is HHDN, then endosulfan.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
11 Li
Measure the peak heights or areas of endosulfan and 4ieWr±n- n
from both the standard-internal standard solution and the sample-
internal standard solution.
-------
Endosulfan EPA-3
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
RF . (wt. HHDN)(% purity HHDN)(pk. ht. or area endosulfan)
(wt. endosulfan)(% purity endosulfan)(pk. ht. or area HHDN)
Determine the percent endosulfan for each injection of the
sample-internal standard solution as follows and calculate the
average:
7 (wt. HHDN)(% purity HHDN)(pk. ht. or area endosulfan)
" (wt. sample) (pk. ht. or area HHDN) (RF) '
This method is based on EPA Experimental Method 62A submitted by
G. Radan, EPA, Region II, New York, N. Y. Some changes and additions
have been made in the write-up; therefore, any comments, criticisms,
suggestions, data, etc. concerning this method will be appreciated.
-------
November 1975
Determination of Endosulfan
by Gas-Liquid Chromatography
(FID - Internal Standard)
Endosulfan EPA-4
(Tentative)
Endosulfan is the accepted common name for hexachlorohexahydro-
methano-2,4,3-benzodioxathiepin-3-oxide, a registered pesticide having
the chemical structure: Q
CH2—0,
CH2—
S=0
Molecular formula: C.H,C1,0,S
y b b J
Molecular weight: 406.9
Melting point: (see below)
Physical state, color, and odor: endosulfan is an odorless white
crystalline solid mixture of two isomers with rap's
of 106°C and 212°C; the technical product is a
brownish crystalline solid, mp 70-100°C, with a
4:1 ratio of the above isomers. Both isomers are
insecticidally active.
Solubility: practically insoluble in water, but soluble in most organic
solvents
Stability: generally quite stable; decomposition catalyzed by iron;
slowly hydrolyzed by water; sensitive to acid and bases;
compatible with non-alkaline pesticides
-------
2 Endosulfan EPA-4
(Tentative)
Other names: Thiodan (Farwerke Hoechst), Beosit, Chlorthiepin, Cyclodan,
Insectophene, Kop-Thiodan, Malix, Thifor, Thimul, Thionex,
HOE 2671, NIA 5A62, FMC 5462
Reagents:
1. Endosulfan standard of known % purity
2. Dieldrin standard of known HEOD content
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.15 gram HEOD into a 50 ml
volumetric flask, dissolve in, and make to volume with acetone.
(cone 3 mg HEOD/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm ID glass column packed with 5% OV-210 on
80/100 mesh Chromosorb W HP (or equivalent column)
3. Precision liquid syringe: 5 or 10 jul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 180°
Injection temperature: 230°
Detector temperature: 230°
Carrier gas: Nitrogen
Carrier gas pressure: 40-60 psi
Hydrogen pressure: 20 psi
Air pressure: 30 psi
-------
3 Endosulfan EPA-4
(Tentative)
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response and
reproducibility.
Procedure:
Preparation of Standard;
Weigh 0.12 gram endosulfan standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 20 ml of the internal
standard solution and shake to dissolve. (final cone 6 mg endo-
sulfan and 3 mg HEOD/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.12 gram endosulfan
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the endosulfan. For
coarse or granular materials, shake mechanically for 30 minutes
or shake by hand intermittently for one hour. (final cone 6 mg
endosulfan and 3 mg HEOD/ml)
Determination:
Inject 2-4 jjl of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is endosulfan, then HEOD
(see note).
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
-------
Endosulfan EPA-4
(Tentative)
Calculation:
Measure the peak heights or areas of endosulfan and HEOD from
both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. HEOD)(% purj.ty HEOD) (pk. ht. or^ area endosulfan)
K r "•" / . t i «- \ / tv * _ _ • , _ t i i- \ / i i. .
(wt. endosulfan) (% purity endosulfan) (ok. ht . or area HEOD)
Determine the percent endosulfan for each injection of the
sample-internal standard solution as follows and calculate the
average:
(wt. HEOD) (% purity HEOD) (pk. ht_._ or area end
__
° " (wt. sample) (pk. ht. or area HEOD) (RF) f(A~l)
Note! Endosulfan consists of two isomers (I and II) which elute
before and after the HEOD. Calculate results using isomer
I (1st peak); however, if results are low, calculate using
the total of isomers I and II (both peaks) . The ratio of
isomers I and II varies considerably among various samples
and standards. Endosulfan II and parathion are not completely
separated on this column, but this does not seem to affect
either the endosulfan II or parathion results significantly.
This method was submitted by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond, Virginia 23219,
This method has been designated as tentative since it is a Va. Exp.
method and because some of the data has been suggested by EPA's
Beltsville Chemistry Lab. Any comments, criticisms, suggestions,
data, etc. concerning this method will be appreciated.
-------
December 1975
Endothall EPA-1
Determination of Endothall in Formulations
(Oxidation and Acid-Base Titration)
Endothall is the accepted common name for 7-oxabicyclo (2.2.1)
heptane-2,3-dicarboxylic acid, a registered herbicide having the
chemical structure:
H2-C
CH—COOH
CH—COOH
Molecular formula: C0H,nOc
o 1U 5
Molecular weight: 186.2
Melting point: 14A°C (some decomposition, see below)
Physical state, color, and odor: white, odorless, crystalline solid
Solubility: solubility in grams per 100 ml at 25°C is: 10 in water,
7 in acetone, 0.1 in benzene, 7.6 in dioxane, 28 in methanol
Stability: stable to light; stable to about 90°C, after which it under-
goes a slow conversion to the anhydride; stable in acid,
non-flammable; non-corrosive to metals
Other names: Endothal (Pennwalt), endothal (Europe except Italy), Accel-
erate, Aquathol, Des-i-cate, Herbicide 273, Herbicide 283,
Hydout, Hydrothol, Tri-Endothal
-------
2 Endothall EPA-1
Principle of the Method:
The sample is neutralized with sulfuric acid (because of residual
sodium hydroxide from manufacturing). It is then evaporated and ashed
to convert the carboxylic acid to carbonate which is determined acidi-
metrically. Salts of carboxylic acids other than endothall interfere.
If ammonium sulfate is present, it must be volatilized.
Reagents:
1. Sodium hydroxide, 0.1N standardized solution
2. Sulfuric acid, 0.1N standardized solution
3. Phenolphthalein indicator solution
4. Sodium hydroxide pellets, ACS
Equipment:
1. Platinum evaporating dish
2. Steam bath and/or drying oven
3. Muffle furnace
4. Filtration apparatus
5. Titration apparatus
6. Usual laboratory glassware
Procedure:
Weigh a portion of sample equivalent to 0.25-0.30 gram endothall
acid (0.31-0.37 gram disodium salt) into a platinum evaporating dish.
Wet dry samples with a few ml water.
(If ammonium sulfate is present, add 1 gram sodium
hydroxide, mix well, and evaporate to dryness.)
-------
Endothall EPA-1
Neutralite carefully with 0.1N Bulfuric acid to just colorless
with phenolphthalein. Evaporate and ash at approx. 525°C. Cool,
extract with hot water, and filter through paper into a 500 ml Erlenmeyer
flask, washing with water. Return the paper to the platinum crucible,
dry, and ash completely. Cool, dissolve the residue in water, and add
to the extract in the Erlenmeyer flask.
Add 50 ml exactly 0.1N sulfurlc acid solution and boll 20 minutes
to remove carbon dioxide. Cool, and titrate with 0.1N sodium hydroxide
solution to the phenolphthalein endpoint.
Calculate the endothall as follows:
z m (ml H2S04)(N H2S04) - (ml NaOH)(N NaOH)(0.0931)(100)
(grams sample)
milliequivalent weight endothall acid » 0.0931
milliequivalent weight endothall, disodium salt « 0.1151
Z endothall acid X 1.236 - Z endothall disodium salt
-------
December 1975
Endothall EPA-2
(Tentative)
Determination of Endothall
by Gas-Liquid Chromatographv (FID)
Endothall is the accepted common name for 7-oxabicvclo (2.2.1)
heptane-2,3-dicarboxylic acid, a registered herbicide having the
chemical structure:
CH—COOH
Ho-C
CH—COOH
Molecular formula: C0H nOc
o 1U j
Molecular weight: 186.2
Melting point: 14A°C (some decomposition, see below)
Physical state, color, and odor: white, odorless, crystalline solid
Solubility: solubility in grams per 100 ml at 25°C is: 10 in water,
7 in acetone, 0.1 in benzene, 7.6 in dioxane, 28 in methanol
Stability: stable to light; stable to about 90°C, after which it under-
goes a slow conversion to the anhydride; stable in acid,
non-flammable; non-corrosive to metals
Other names: Endothal (Pennwalt), endothal (Europe except Italy), Accel-
erate, Aquathol, Des-i-cate, Herbicide 273, Herbicide 283,
Hydout, Hydrothol, Tri-Endothal
This method applies to the salts of endothall, e.g.,mono (N,N-dimethyl-
alklamine salt) as well as to the free acid.
-------
2 Endothall EPA-2
(Tentative)
Reagents:
1. Endothall standard of known % purity
2. Acetonitrile, pesticide or spectro grade
3. 3M Sulfuric acid, ACS
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 5' x 1/4" O.D. glass, packed with 3% SE-30 on
60/80 Chromosorb W AW DMCS (or equivalent column)
3. Precision liquid syringe: 10 ^il
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 130°C
Injection temperature: 180°C
Detector temperature: 180°C
Carrier gas: Nitrogen
Carrier gas flow rate: 40 ml/min
Hydrogen flow rate: adjusted for specific GC
Air flow rate: adjusted for specific GC
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Endothall EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.075 gram endothall standard into a small glass-stoppered
flask or screw-cap bottle, add 8 drops 3M sulfuric acid, 25 ml
acetonitrile by pipette, and shake to dissolve. (cone 3 ug/iil)
Preparation of Sample:
For technical material and liquid formulations, weigh a portion
of sample equivalent to 0.075 gram endothall into a 25 ml volumetric
flask, add 8 drops 3M sulfuric acid, make to volume with aceto-
nitrile, and mix thoroughly. (final cone 3 jig endothall/jal)
For dry formulations, weigh a portion of sample equivalent to
0.150 gram of endothall into a 125 ml screw-cap flask, add 8 drops
3M sulfuric acid, 50 ml acetonitrile by pipette, and shake for one
hour. Allow to settle; filter or centrifuge if necessary taking
precautions to prevent evaporation. (final cone 3 ug endothall/jul)
Determination:
Using a precision liquid syringe, alternately inject three
5 /il portions each of standard and sample solutions. Measure the
peak height or peak area for each peak and calculate the average
for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
-------
Endothall EPA-2
(Tentative)
Calculation:
From the average peak height or peak area calculate the
percent endothall as follows:
«, ^ (pk. ht. or area sample)(wt. std injected) (% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Florida Department of Agriculture and Consumer
Services, Mayo Building, Tallahassee, Florida 32304.
This method has been designated as tentative since some data has been
suggested by EPA's Beltsville Chemistry Laboratory. Any comments,
criticisms, suggestions, data, etc. concerning this method will be
appreciated.
-------
October 1975 EPIC EPA-1
(Tentative)
Determination of EPTC by
Gas-Liquid Chromatography
(TCD - Internal Standard)
EPTC is the common name for S-ethyl dipropylthiocarbamate, a
registered herbicide having the chemical structure:
0
|| ,CH2—CH2—CH3
CH3 CH2 S C N <^
— CH2—CH3
Molecular formula: C H NOS
Molecular weight: 189.3
Boiling point: 127°C at 20 mm Hg (235°C by extrapolation)
Physical state, color, and odor: Light yellow-colored liquid with
an amine odor
Solubility: 365 ppm in water at 20°C; miscible with acetone,
benzene, ethanol, isopropanol, kerosene, methanol,
methyl isobutyl ketone, toluene, and xylene
Stability: stable, non-corrosive
Other names: Eptam (Stauffer), Eradicane, Knoxweed
Reagents;
1. EPTC standard of known % purity
2. 2-ethyl-l,3-hexanediol standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.5 gram ethyl hexanediol
into a 50 ml volumetric flask, dissolve in, and make to volume
with acetone. (cone 10 mg/ml)
-------
EPIC EPA-1
(Tentative)
Equipment;
1. Gas chromatograph with thermal conductivity detector (TCD)
2. 6' x 1/4" glass column packed with 10% SE-30 on 100/120
mesh Diatoport S (or equivalent column)
3. Precision liquid syringe - 5 or 10 ul
4. Usual laboratory glassware
Operating Conditions for TCP:
Column temperature: 130°C
Injection temperature: 225°C
Detector temperature: 150°C
Filament current: 200 ma
Carrier gas: Helium
Flow rate: 30 ml/min
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.1 gram EPIC standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 10 ml of the internal
standard solution and shake to dissolve. (final cone 10 mg EPIC
and 10 mg ethyl hexanediol/ml)
-------
3 EPIC EPA-1
(Tentative)
Preparation of Sample^
Weigh a portion of sample equivalent to 0.25 gram EPTC into
a small glass-stoppered flask or screw-cap bottle. Add by
pipette 25 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the EPTC. For
coarse or granular materials, shake mechanically for 10-15
minutes or shake by hand intermittently for 25-30 minutes.
(final cone 10 rag EPTC and 10 mg ethyl hexanediol/ml)
Determination:
Inject 2-3 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within approx. 10 minutes and peak heights of from
1/2 to 3/4 full scale. The elution time of ethyl hexanediol is
approx. 2 minutes and that of EPTC approx. 4 minutes.
Proceed with the determination, making at least three
injections each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of EPTC and ethyl hexanediol
from both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
IS = internal standard = ethyl hexanediol
= (wt. IS)(% purity IS)(pk. ht. or area EPTC)
(wt. EPTC)(% purity EPTC)(pk. ht. or area IS)
Determine the percent EPTC for each injection of the sample-
internal standard solution as follows and calculate the average:
% = (wt. x % purity IS)(pk. ht. or area EPTC)
(wt. sample) (pk. ht. or area IS) (RF)
Method submitted by George Radan, EPA Region II, New York, N. Y.
-------
July 1975 EPIC EPA-2
(Tentative)
Determination of EPTC by
High Pressure Liquid Chromatography
'' >' '1'
EPTC is the common name for S-ethyl dipropylthiocarbamate, a
registered herbicide having the chemical structure:
0
|| CH2 —CH2—CH3
CH3—CH2—S—C N<^
XH2— CH2—CH3
Molecular formula: C-H ^NOS
Molecular weight: 189.3
Boiling point: 127°C at 20 mm Hg (235°C by extrapolation)
Physical state, color, and odor: Light yellow-colored liquid with
an amine odor
Solubility: 365 ppm in water at 20°C; miscible with acetone,
benzene, ethanol, isopropanol, kerosene, methanol,
methyl isobutyl ketone, toluene, and xylene
Stability: stable, non-corrosive
Other names: Eptam (Stauffer), Eradicane, Knoxweed
Reagents;
1. EPTC standard of known % purity
2. Chloroform
3. Dichloromethane
A. Hexane
5. Methanol
All solvents should be pesticide or spectro grade.
-------
2 EPIC EPA-2
(Tentative)
Equipment:
1. High pressure liquid chromatograph
2. High pressure liquid syringe or sample injection loop
3. Liquid chromatographic column, 4 mm I.D. x 25 cm packed
with LiChrosorb Si 60 - 10 ji (or equivalent column)
4. Usual laboratory glassware
Operating conditions for Hewlett-Packard Model 1010B LC;
Mobile phase: 40 ml methanol in 2000 ml of a mixture containing
80% dichloromethane and 20% hexane
Column temperature: ambient
2
Observed column pressure: 30 kg/cm (425 PSI)
Flow rate: 3 ml/min
Detector: UV at 240 nm
Chart speed: 0.5 in/min
Injection: 10 ul
Conditions may have to be varied by the analyst for other instru-
ments, column variations, sample composition, etc. to obtain optimum
response and reproducibility.
Procedure;
Preparation of Standard:
Weigh 0.02 gram EPIC standard into a 10 ml volumetric
flask; dissolve and make to volume with chloroform (final
cone 2 mg/ml).
-------
3 EPIC EPA-2
(Tentative)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.2 gram EPIC
into a 100 ml volumetric flask, make to volume with chloro-
form, and mix thoroughly (final cone 2 mg EPIC/ml).
Determination;
Alternately inject three 10 jil portions each of standard
and sample solutions. Measure the peak height or peak area for
each peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent EPIC as follows:
7 „ (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method developed by Joseph B. Audino, Supervisor, Pesticide Formulation
Laboratory, California Department of Food and Agriculture; and by
Yoshihiko Kawano, Associate Chemist on sabbatical leave from the
University of Hawaii.
-------
October 1975 EPIC EPA-3
Determination of EPIC by
Gas-Liquid Chromatography
(FID - Internal Standard)
EPTC is the common name for S-ethyl dipropylthiocarbamate, a
registered herbicide having the chemical structure:
0
|| .xCH2—CH2—CH3
CH^— CH2— S— C N
^ -CH2—CH2—CH3
Molecular formula: C H NOS
Molecular weight: 189.3
Boiling point: 127°C at 20 mm Hg (235°C by extrapolation)
Physical state, color, and odor: Light yellow-colored liquid with
an amine odor
Solubility: 365 ppm in water at 20°C; miscible with acetone, benzene,
ethanol, isopropanol, kerosene, methanol, methyl isobutyl
ketone, toluene, and xylene
Stability: stable, non-corrosive
Other names: Eptam (Stauffer), Eradicane, Knoxweed
Reagents:
1. EPTC standard of known % purity
2. Butylate standard of known % purity
3. Carbon disulfide, pesticide or spectro grade
-------
2 EPIC EPA-3
Reagents (Cont.)
4. Chloroform, pesticide or spectro grade
5. Hethanol, pesticide or spectro grade
6. Internal Standard solution - weigh 0.25 gram butylate into
a 50 ml volumetric flask, dissolve in, and make to volume
with a solvent mixture consisting of 80% carbon disulfide
+ 15% chloroform + 5% methanol. (cone 5 mg butylate/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6" x 2 mm glass column packed with 3% OV-1 on
60/80 Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 pi
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 120°C
Injection temperature: 225°C
Detector temperature: 250°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi
Hydrogen pressure: 20 psi
Air pressure: 30 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
3 EPIC EPA-3
Procedure:
Preparation of Standard:
Weigh 0.08 gram EPTC standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 20 ml of the internal
standard solution and shake to dissolve. (final cone 4 mg EPTC
and 5 mg butylate/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.08 gram EPTC into
a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the EPTC. For
coarse or granular materials, shake mechanically for 10-15
minutes or shake by hand intermittently for 25-30 minutes.
(final cone 4 mg EPTC and 5 mg butylate/ml)
Pet ermina t ion:
Inject 2-3 pi of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from
1/2 to 3/4 full scale. The elution order is EPTC 1st and butylate
2nd.
Proceed with the determination, making at least three
injections each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of EPTC and butylate from
both the standard-internal standard solution and the sample-
internal standard solution.
-------
4 EPIC EPA-3
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
_ (wt. butylate)(% purity butylate) (pk. ht . or area EPIC)
(wt. EPTC)(% purity EPTC)(pk. ht. or area butylate)
Determine the percent EPTC for each injection of the sample
internal standard solution as follows and calculate the average
(wt. butylate) (% purity butylate) (pk. ht. or area EPIC) (
(wt. sample) (pk. ht. or area butylate) (RF)
Method submitted by Division of Regulatory Services, Kentucky
Agricultural Experiment Station, University of Kentucky, Lexington,
Kentucky 40506.
(See EPA-4 for a similar method submitted by Virginia State Laboratories.)
-------
October 1975 EPIC EPA-A
(Tentative)
Determination of EPIC by
Gas-Liquid Chromatography
(FID - Internal Standard)
EPIC is the common name for S-ethyl dipropylthiocarbamate, a
registered herbicide having the chemical structure:
|| CH2—CH2—CH3
CH3 —CH2—S—C N <^
XH2—CH2—CH3
Molecular formula: C H._NOS
Molecular weight: 189.3
Boiling point: 127°C at 20 mm Hg (235°C by extrapolation)
Physical state, color, and odor: Light yellow-colored liquid with
an amine odor
Solubility: 365 ppm in water at 20°C; miscible with acetone, benzene,
ethanol, isopropanol, kerosene, methanol, methyl isobutyl
ketone, toluene, and xylene
Stability: stable, non-corrosive
Other names: Eptam (Stauffer), Eradicane, Knoxweed
-------
2 EPIC EPA-A
(Tentative)
Reagents:
1. EPIC standard of known % purity
2. Vernolate standard of known % purity
3. Carbon disulfide, pesticide or spectro grade
4. Chloroform, pesticide or spectro grade
5. Methanol, pesticide or spectro grade
6. Internal Standard solution - weigh 0.2 gram vernolate into
a 50 ml volumetric flask, dissolve in, and make to volume
with a solvent mixture consisting of 80% carbon disulfide
•f 15% chloroform + 5% methanol. (cone 4 mg vernolate/ml)
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm ID glass column packed with 5% SE-30
on 80/100 Chromosorb W HP (or equivalent column)
3. Precision liquid syringe: 5 or 10 pi
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 130°
Injection temperature: 200°
Detector temperature: 200°
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi
Hydrogen pressure: 20 psi
Air pressure: 30 psi
-------
3 EPIC EPA-4
(Tentative)
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.05 gram EPTC standard into a small glass-stoppered
flask or screw-cap tube. Add by pipette 20 ml of the internal
standard solution and shake to dissolve. (final cone 2.5 rag
EPTC and 4 mg vernolate/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.05 gram EPTC
into a small glass-stoppered flask or screw-cap tube. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the EPTC. For
coarse or granular materials, shake or tumble mechanically for
30 minutes or shake by hand intermittently for one hour.
(final cone 2.5 mg EPTC and 4 mg vernolate/ml)
Determination:
Inject 1-2 p.1 of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is EPTC, then vernolate.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
-------
EPIC EPA-4
(Tentative)
Calculation:
Measure the peak heights or areas of EPIC and vernolate
from both the standard-internal standard solution and the
sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
RF _ (wt. vernolate) (% purity vernolate) (pk. ht. or area EPTC)
(wt. EPTC)(% purity EPTC)(pk. ht. or area vernolate)
Determine the percent EPTC for each injection of the sample-
internal standard solution as follows and calculate the average:
(wt. vernolate) (% purity vernolate) (pk. ht. or area EPTC) (j
(wt. sample) (pk. ht. or area vernolate) (RF) ' L4- \
This method was submitted by the Commonwealth of Virginia, Division
of Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
Note! This method has been designated as tentative since it is a
Va. Exp. method and because some of the data has been suggested
by EPA's Beltsville Chemistry Lab. Any comments, criticism,
suggestion, data, etc. concerning this method will be appreciated,
-------
October 1975 EPIC EPA-5
(Tentative)
Determination of EPTC by
Gas—Liquid Chromatography
(TCD - Internal Standard)
EPTC is the common name for S-ethyl dipropylthiocarbamate, a
registered herbicide having the chemical structure:
0
|| /CH2—CH2—CH3
CH3 CH2 S—C N <^
XIH2—-CH2
Molecular formula: C H NOS
Molecular weight: 189.3
Boiling point: 127°C at 20 mm Hg (2358C by extrapolation)
Physical state, color, and odor: Light yellow-colored liquid with
an amine odor
Solubility: 365 ppm in water at 20°C; miscible with acetone, benzene,
ethanol, isopropanol, kerosene, methanol, methyl isobutyl
ketone, toluene, and xylene
Stability: stable, non-corrosive
Other names: Eptam (Stauffer), Eradicane, Knoxweed
-------
2 EPIC EPA-5
(Tentative)
Reagents:
1. EPIC standard of known % purity
2. Vernolate standard of known Z purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.25 gram vernolate into
a 25 ml volumetric flask, dissolve in, and make to volume
with acetone. (cone 10 mg vernolate/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 5' x 1/4" glass column packed with 5% PEG-1540 on
60/80 Chromosorb W AW DMCS (or equivalent column)
3. Precision liquid syringe: 25 or 50 pi
4. Usual laboratory glassware
Operating Conditions for TCD:
Column temperature: 160°C
Injection temperature: 200°C
Detector temperature: 200°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas flow rate: 30 ral/min
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
EPIC EPA-5
(Tentative)
Procedure:
Preparation of Standard^
Weigh 0.08 gram EPIC standard into a small glass-stoppered
flask or screw-cap tube. Add by pipette 10 ml of the internal
standard solution and shake to dissolve. (final cone 8 mg
EPIC and 10 mg vernolate/ml)
of Sample:
Weigh a portion of sample equivalent to 0.08 gram EPIC
into a small glass-stoppered flask or screw-cap tube. Add by
pipette 10 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the EPTC. For
coarse or granular materials, shake or tumble mechanically for
30 minutes or shake by hand intermittently for one hour.
(final cone 8 mg EPTC and 10 mg vernolate/ml)
Determination:
Inject 10-15 pi of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from
1/2 to 3/4 full scale. The elution order is EPTC, then vernolate.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of EPTC and vernolate from
both the standard-internal standard solution and the sample-
internal standard solution.
-------
EPIC EPA-5
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. vernolate)(% purity vernolate)(pk. ht. or area EPIC)
x (wt. EPTC)(% purity EPTC)(pk. ht. or area vernolate)
Determine the percent EPTC for each injection of the
sample-internal standard solution as follows and calculate the
average:
_ (wt. vernolate) (% purity vernolate) (pk. ht. or area EPTC)f3r6»)-
(wt. sample) (pk. ht. or area vernolate) (RF) ;LA~\]
This method was submitted by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
Note! This method has been designated as tentative since it is a
Va. Exp. method and because some of the data has been suggested
by EPA's Beltsville Lab. Any comments, criticism, suggestion,
data, etc. concerning this method will be appreciated.
-------
September 1975 Ethion EPA-1
Determination of Ethion in Solid
Formulations by Infrared Spectroscopy
Ethion is the accepted common name for 0,0,0',O'-tetraethyl
S,S'-methylene bisphosphorodithioate, a registered insecticide
having the chemical structure:
-CH2-0 S s o-CH2-CH3
V—S—CH2—S P/
CH3—CH2—0 \__CH2 — CH3
Molecular formula: CJH 0 P2S,
Molecular weight: 384.48
Boiling point: 164 to 165°C at 0.3 mm Hg; solidifies at -12 to
-15°C
Physical state, color, and odor: pure form is an odorless, colorless
liquid; technical product is a yellow to amber
liquid
Solubility: very slightly soluble in water; poorly soluble in
aliphatic solvents; highly soluble in aromatic solvents
Stability: slowly oxidizes in air; subject to hydrolysis by both
acids and alkalis
Other names: NIA 1240 and Nialate (FMC Corp.), diethion, Ethodan
-------
2 Ethion EPA-1
Reagents;
1. Ethion standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm NaCl or KBr cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure;
Preparation of Standard;
Weigh 0.10 gram ethion standard into a 10 ml volumetric
flask; make to volume with carbon disulfide. Add a small amount
of anhydrous sodium sulfate to insure dryness. (final cone
10 mg/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.40 gram ethion
into a glass-stoppered flask or screw-cap tube. Add 100 ml
carbon disulfide by pipette and 1-2 grams anhydrous sodium sulfate.
-------
3 Ethion EPA-1
Close tightly and shake for one hour. Allow to settle; centrifuge
of filter if necessary, taking precaution to prevent evaporation.
Evaporate a 25 ml aliquot to about 5 ml, transfer to a 10 ml
volumetric flask, and make to volume with carbon disulfide. Add
a small amount of anhydrous sodium sulfate to insure dryness.
(final cone 10 mg ethion/ml)
*
Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan both the standard and sample from
714 cm"1 to 595 cm"1 (14.0 ;i to 16.8 p).
Determine the absorbance of standard and sample using the
peak at 647 cm (15.45 ji) and baseline from 701 cm to 615 cm
(14.25 fi to 16.25 fi).
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent ethion as follows:
% . (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in rag/ml)
(A concentration of 1 mg ethion/ml carbon disulfide gives an
absorbance of approx. 0.04 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services.
* Eva Santos, EPA Region IX, San Francisco, California, has contributed
a similar method using:
958 cm (10.44 ji) analytical absorption band
981 cm" (10.19 ji) basepoint
-------
October 1975 Ethion EPA-2
(Tentative)
Determination of Ethion by
Gas-Liquid Chromatography (TCD)
Ethion is the accepted common name for 0,0,0',O'-tetraethyl
S,S'-methylene bisphosphorodithioate, a registered insecticide having
the chemical structure:
CH3—CH2—0 S S o—CH2—CH3
V—S — CH2 — S P<^
CH3—CH2—0 \__CH2 —CH3
Molecular formula: CHOPS,
Molecular weight: 384.48
Boiling point: 164 to 165°C at 0.3 mm Hg; solidifies at -12 to
-15°C
Physical state, color, and odor: pure form is an odorless, colorless
liquid; technical product is a yellow to amber
liquid
Solubility: very slightly soluble in water; poorly soluble in
aliphatic solvents; highly soluble in aromatic solvents
Stability: slowly oxidizes in air; subject to hydrolysis by both
acids and alkalis
Other names: NIA 1240 and Nialate (FMC Corp.), diethion, Ethodan
Reagents:
1. Ethion standard of known % purity
2. Chloroform, pesticide or spectro grade
-------
2 Ethion EPA-2
(Tentative)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 5' x 1/4" glass column packed with 10% QF-1 on
Chromosorb W, AW, DMCS (or equivalent column)
3. Precision liquid syringe: 50 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for TCP;
Column temperature: 210°C
Injection temperature: 240°C
Detector temperature: 240°C
Filament current: 200 ma
Carrier gas: Helium
Flow rate: 100 ml/min
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response and
reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.2 gram ethion standard into a 10 ml volumetric flask;
dissolve and make to volume with chloroform, (final cone 20 mg/ml)
-------
3 Ethion EPA-2
(Tentative)
Preparation of Sample:
For technical material and liquid formulations, weigh a
portion of sample equivalent to 0.20 gram ethion into a 10 ml
volumetric flask, make to volume with chloroform,and mix
thoroughly. (final cone 20 mg ethion/ml)
For dry formulations, weigh a portion of sample equivalent
to 1.0 gram ethion into a glass-stoppered flask or screw-cap
bottle, add by pipette 50 ml chloroform, close tightly, and
shake for one hour. Allow to settle; filter or centrifuge if
necessary, taking precautions to prevent evaporation. (final
cone 20 mg ethion/ml)
Determination;
Using a precision liquid syringe, alternately inject three
30-40 ul portions each of standard and sample solutions. Measure
the peak height or peak area for each peak and calculate the
average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent ethion as follows:
7 _ (pk. ht. or area sample)(wt. std injected)(% purity std)
(pk. ht. or area std)(wt. sample injected)
Method submitted by Eva Santos, EPA Region IX, San Francisco,
California.
-------
December 1975 Ethoprop EPA-1
(Tentative)
Determination of Ethoprop
by Infrared Spectroscopy
Ethoprop is a common name for 0-ethyl-S,S-dipropyl phosphoro-
dithioate, a registered nematocide and soil insecticide having the
chemical structure:
CH2—CH2—CH3
II ^
CH3—CH2-
\
CH2—CH2—CH3
Molecular formula: CQHiQ°2PS2
Molecular weight: 242.3
Melting point: 86 to 91°C at 0.2 mm Hg
Physical state, color, and odor: clear yellowish liquid with a strong
mercaptan odor
Solubility: insoluble in water; soluble in most organic solvents
Stability: very stable in acid aqueous media from 25 to 100°C;
hydrolyzed in basic media moderately fast at 25°C and
rapidly at 100°C; thermal stability is good for 8 hours
at 150°C
Other names: Mocap (Mobil), prophos (discontinued because of conflict),
VC 9-104
-------
2 Ethoprop EPA-1
(Tentative)
Reagents:
1. Ethoprop standard of known % purity
2. Chloroform, pesticide or spectre grade
3. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording,
with matched 0.2 mm NaCl or KBr cells
2. Mechanical shaker
3. Filtration apparatus or centrifuge
A. Usual laboratory glassware
Procedure;
Preparation of Standard;
Weigh 0.1 gram ethoprop into a small glass-stoppered flask
or screw-cap bottle, add 10 ml chloroform by pipette, and shake
to dissolve. Add a small amount of anhydrous sodium sulfate to
insure dryness. (final cone 10 mg/ml)
Preparation of Sample;
For dusts, granules, and wettable powders, weigh a portion of
sample equivalent to 0.5 gram ethoprop into a 125 ml glass-
stoppered or screw-cap Erlenmeyer flask. Add 50 ml chloroform by
pipette and 1-2 grams anhydrous sodium sulfate. Close tightly,
shake on a mechanical shaker for 1 hour, allow to settle, filter
or centrifuge if necessary, taking precaution to avoid evaporation.
(final cone 10 mg ethoprop/ml)
-------
3 Ethoprop EPA-0
(Tentative)
For emulsiftable concentrates, weigh a portion of sample
equivalent to 0.5 gram ethoprop into a 125 ml glass-stoppered
flask or screw-cap bottle. Add 50 ml chloroform by pipette and
sufficient anhydrous sodium sulfate to clarify the solution (after
shaking). Close tightly and shake vigorously on a mechanical
shaker for one hour. Allow to settle; filter or centrifuge if
necessary to get a clear chloroform solution, taking precaution
to prevent evaporation, (final cone 10 mg ethoprop/ml)
(There may be interference from the emulsifier in the
sample; if so, another procedure must be used.)
Determinat ion r
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1100 cm to
900 cm"1 (9.1 ju to 11.1 ^) .
Determine the absorbance of standard and sample using the peak
at 1012 cm (9.9 p) and baseline from 1070 cm to 970 cm
(9.35 p to 10.3 ja).
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent ethoprop as follows:
„ m (abs. sample)(cone, std in mg/ml)(Z purity std)
(abs. std)(cone, sample in mg/ml)
Method submitted by Mark W. Law, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
Any criticisms, suggestions, data, etc. on the use of this method will
be appreciated.
-------
December 1975 Ethoprop EPA-2
(Tentative)
Determination of Ethoprop
by Gas-Liquid Chromatography
(TCD - Internal Standard)
Ethoprop is a common name for 0-ethyl-S,S-dipropyl phosphoro-
dithioate, a registered neraatocide and soil insecticide having the
chemical structure:
S—CH2 — CH2—CH3
CH3 — CH2 — 0 - P
CH2— CH2-CH3
Molecular formula: C0H100,PS.
o iy / 2.
Molecular weight: 242.3
Melting point: 86 to 91°C at 0.2 mm Hg
Physical state, color, and odor: clear yellowish liquid with a strong
mercaptan odor
Solubility: insoluble in water; soluble in most organic solvents
Stability: • very stable in acid aqueous media from 25 to 100°C;
hydrolyzed in basic media moderately fast at 25°C and
rapidly at 100°C; thermal stability is good for 8 hours
at 150°C
Other names: Mocap (Mobil), prophos (discontinued because of conflict),
VC 9-104
-------
2 Ethoprop EPA-2
(Tentative)
Reagents;
1. Ethoprop standard of known % purity
2. Diazinon standard of known % purity
3. Chloroform, pesticide or spectro grade
4. Internal Standard solution - weigh 0.5 gram diazinon into a
50 ml volumetric flask and make to volume with chloroform.
(cone 10 mg diazinon/ml)
Equipment;
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 6' x 1/8" I.D. SS packed with 10% SE 30 on 60/80
Diatoport S (or equivalent column)
3. Precision liquid syringe: 10 ul
4. Usual laboratory glassware
Operating Conditions for TCD;
Column temperature: 200°C
Injection temperature: 225°C
Detector temperature: 225°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas flow: adjusted for particular GC
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response and
reproducibility.
-------
Ethoprop EPA-2
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.16 gran ethoprop standard Into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 20 •! of
the internal standard solution and shake to dissolve, (final
cone 8 ng ethoprop and 10 ng diazinon/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.16 gram ethoprop
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the ethoprop. For
coarse or granular materials, shake mechanically for 30 minutes
or shake by hand intermittently for one hour, (final cone 8 mg
ethoprop and 10 mg diazinon/ml)
Determination;
Inject 1-2 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is ethoprop, then diazinon.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of ethoprop and diazinon
from both the standard-internal standard solution and the sample-
internal standard solution.
-------
Ethoprop EPA-2
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. diazinon)(% purity diazinon)(pk. ht. or area ethoprop)
(wt. ethoprop)(% purity ethoprop)(pk. ht. or area diazonon)
Determine the percent ethoprop for each injection of the
sample-internal standard solution as follows and calculate the
average:
_ m (wt. diazinon)(% purity diazinon)(pk. ht. or area ethoprop)
" * (wt. sample)(pk. ht. or area diazinon)(RF)
Method submitted by Stelios Gerazounis, EPA Region II, New York, N. Y.
This method was designated as EPA Experimental Method No. 34 and was
based on data from the Virginia Department of Agriculture. Some changes
have been made and data added in this write-up; therefore, any comments,
criticisms, suggestions, data, etc. concerning this method will be
appreciated.
-------
December 1975 Ethoprop EPA-3
(Tentative)
Determination of Ethoprop
by Gas-Liquid Chromatography
(FID - Internal Standard)
Ethoprop is a common name for 0-ethyl-S,S-dipropyl phosphoro-
dithioate, a registered nematocide and soil insecticide having the
chemical structure:
S — CH2 —CH2 —CH3
CH3—CH2—0 P:
— CH2—CH2 — CH3
Molecular formula: C0H,00_PS.
o iy 2. 2.
Molecular weight: 242.3
Melting point: 86 to 91°C at 0.2 mm Hg
Physical state, color, and odor: clear yellowish liquid with a strong
tnercaptan odor
Solubility: insoluble in water; soluble in most organic solvents
Stability: very stable in acid aqueous media from 25 to 100°C;
hydrolyzed in basic media moderately fast at 25°C and
rapidly at 100°C; thermal stability is good for 8 hours
at 150°C
Other names: Mocap (Mobil), prophos (discontinued because of conflict),
VC 9-104
-------
2 Ethoprop EPA-3
(Tentative)
Reagents;
1. Ethoprop standard of known % purity
2. Diazinon standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.25 gram diazinon into a
50 ml volumetric flask and make to volume with acetone.
(cone 5 mg diazinon/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm ID glass column packed with 5% SE-30 on
80/100 Chromosorb W HP (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 180°C
Injection temperature: 230°C
Detector temperature: 230°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi (adjust for specific GC)
Hydrogen pressure: 20 psi (adjust for specific GC)
Air pressure: 30 psi (adjust for specific GC)
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response and
reproducibility.
-------
Ethoprop EPA-3
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.06 gram ethoprop standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 20 ml of the internal
standard solution and shake to dissolve, (final cone 3 mg ethoprop
and 5 mg diazinon/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.06 gram ethoprop
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the ethoprop. For
coarse or granular materials, shake mechanically for 30 minutes
or shake by hand intermittently for one hour, (final cone 3 mg
ethoprop and 5 mg diazinon/ml)
Determination;
Inject 1-2 jil of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is ethoprop, then diazinon.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of ethoprop and diazinon from
both the standard-internal standard solution and the sample-internal
standard solution.
-------
Ethoprop EPA-3
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. diazinon)(% purity diazinon)(pk. ht. or area ethoprop)
(wt. ethoprop)(% purity ethoprop)(pk. ht. or area diazinon)
Determine the percent ethoprop for each injection of the
sample-internal standard solution as follows and calculate the
average:
„ m (wt. diazinon) (% purity diazinonHpk. ht. or area ethoprop)
(wt. sample) (pk. ht. or area diazinon) (RF) ((4-1)
Method submitted by Commonwealth of Virginia, Division of Consolidated
Laboratory Services, 1 North 14th Street, Richmond, Virginia 23219.
This method has been designated as tentative since it is based on an
experimental method from Virginia.
Variable results are sometimes obtained on duplicate runs of 10% granular
formulations, probably due to the small sample size used and the non-
uniform size of the granules. A larger sample with corresponding increase
in internal standard solution solvent or a Soxhlet extraction may be
necessary.
-------
January 1976 Ethyl Hexanediol EPA-1
Determination of Ethyl Hexanediol
by Acetylation and Titration
Ethyl hexanediol is a common name (Ent. Soc. Am.) for 2-ethyl-
1,3-hexanediol, a registered insect repellent having the chemical
structure:
CH2—CH3
CH2—CH CH CH2—CH2—CH3
OH OH
Molecular formula: C_H,000
o lo Z
Molecular weight: 146.2
Boiling point: 244°C; the technical product has a distillation
range of 240 to 250°C
Physical state, color, and odor: colorless liquid; the technical product
has a faint odor of witch hazel
Solubility: slightly soluble in water; miscible with alcohol, chloroform,
ether; will not dissolve nylon, rayon
Stability: stable under normal conditions; both hydroxyl groups can be
esterified, the secondary group with difficulty; it is
without chemical or solvent action on clothing and most
plastics
Other names: ethohexadiol (USP), Rutgers 6-12, 6-12 Insect Repellent,
ethyhexylene glycol
-------
2 Ethyl Hexanediol EPA-1
Principle of the Method:
A known amount of acetic anhydride is reacted with the hydroxyl
groups of ethyl hexanediol and the excess is titrated with sodium
hydroxide.
This method will determine the hydroxyl groups in alcohols, glycols,
and phenols and the amino groups in primary and secondary amines. If
any of these substances are present, they must be removed prior to
analysis. Water, except in very small amounts, interferes by reacting
with the acetylating reagent.
Reagents:
1. Acetic anhydride, ACS
2. Pyridine, ACS, preferably freshly redistilled
3. Acetylating reagent - mix 25 ml acetic anhydride with 75 ml
pyridine
4. Mixed indicator - mix one part 0.1% neutral (to NaOH) cresol
red with 3 parts 0.1% neutral (to NaOH) thymol blue
5. Alcohol sodium hydroxide, 0.5N standardized solution - prepare
from 50% sodium hydroxide solution and aldehyde-free ethanol
(or methanol)
Equipment:
1. Iodine flasks, 300 ml
2. Steam bath
3. Titration apparatus
4. Usual laboratory glassware
-------
3 Ethyl Hexanediol EPA-1
Procedure:
Weigh a portion of sample equivalent to 0.7 gram ethyl hexanediol
into a 300 ml iodine flask, add exactly 10 ml acetylating reagent by
pipette, stopper the flask, and add 1-2 ml pyridine to the well around
the stopper. Add 10 ml acetylating reagent to a second flask for a
blank, and treat exactly as the sample.
Heat the flasks on a steam bath for at least one hour, using the
maximum heat that is practical. Cool; add 10 ml water to the well of
the flask,allowing it to wash down the sides of the loosened stopper
and flask. Mix thoroughly to bring the water into contact with all of
the acetylating reagent.
Add a few drops of the mixed indicator and titrate with 0.5N
alcohol sodium hydroxide solution to a blue endpoint.
Calculation;
Calculate the percent ethyl hexanediol as follows:
% _ (ml NaOH for blank - ml NaOH for sample)(N NaOH)(0.07311)(100)
(grams sample)
0.07311 » milliequivalent weight ethyl hexanediol
-------
January 1976 Ethyl Hexanediol EPA-2
(Tentative)
Determination of Ethyl Hexanediol
by Gas-Liquid Chromatography
(TCD - Internal Standard)
Ethyl hexanediol is a common name (Ent. Soc. Am.) for 2-ethyl-
1,3-hexanediol, a registered insect repellent having the chemical
structure:
CH2—CH3
CH2 —CH CH CH2—CHz—CHj
OH OH
Molecular formula: C0H 00.
o lo /
Molecular weight: 146.2
Boiling point: 244°C; the technical product has a distillation
range of 240 to 250°C
Physical state, color, and odor: colorless liquid; the technical product
has a faint odor of witch hazel
Solubility: slightlv soluble in water; miscible with alcohol, chloroform,
ether; will not dissolve nylon, rayon
Stability: stable under normal conditions; both hydroxyl groups can be
esterified, the secondary group with difficulty; it is
without chemical or solvent action on clothing and most
plastics
Other names: ethohexadiol (USP), Rutgers 6-12, 6-12 Insect Repellent,
ethyhexylene glycol
-------
2 Ethyl Hexanediol EPA-2
(Tentative)
Reagents:
1. 2-Ethyl-l,3-hexanediol of known % purity
2. o-Dichlorobenzene, commercial grade or better
3. Isopropanol, pesticide or spectro grade
4. Internal Standard solution - weigh 0.5 gram o-dichlorobenzene
into a 50 ml volumetric flask; dissolve in and make to volume
with isopropanol. (cone 10 mg o-dichlorobenzene/ml)
Equipment;
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 6' x 1/8" stainless steel, packed with 10% SE-30 on
Diatoport S (or equivalent column)
3. Precision liquid syringe: 10 pi
4. Usual laboratory glassware
Operating Conditions for TCP:
Column temperature: 120°C
Injection temperature: 150°C
Detector temperature: 150°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas pressure: 40 psi (adjust for specific GC)
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Ethyl Hexanediol EPA-2
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.25 gram ethyl hexanediol standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 10 ml of the
internal standard solution and shake to dissolve. (final cone
25 mg ethyl hexanediol and 10 mg o-dichlorobenzene/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.25 gram ethyl hexane-
diol into a small glass-stoppered flask or screw-cap bottle. Add
by pipette 10 ml of the internal standard solution. Close tightly
and shake thoroughly. (final cone 25 mg ethyl hexanediol and 10 mg
o-dichlorobenzene/ml)
De t ermina t ion;
Inject 1 ul of standard and, if necessary, adjust the instrument
parameters and the volume injected to give a complete separation
within a reasonable time and peak heights of from 1/2 to 3/4 full
scale. The elution order is o-dichlorobenzene, then ethyl hexane-
diol.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of ethyl hexanediol and o-
dichlorobenzene from both the standard-internal standard solution
and the sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
-------
Ethyl Hexanediol EPA-2
(Tentative)
I.S. - Internal Standard = o-dichlorobenzene
op « (wt. I. S.) (% purity I.S.) (pk. ht_._ or area ethyj^_hexanediol)
(wt. ethyl hexanediol)(% purity ethyl hexanediol)(pk. ht. or area I.S.)
Determine the percent ethyl hexanediol for each injection of the
sample-internal standard solution as follows and calculate the average:
% = (wt. I.S.)(% purity I.S.)(pk. ht. or area ethyl hexanediol)
(wt. sample)(pk. ht. or area l.S.)(RF) A-/-/\
Note: a 1/4" column can be used at 130°C with very similar results.
Method submitted by George Radan, EPA, Region II, New York, N. Y.
This method is designated as tentative since it is based on EPA's
Experimental Method No. 35 and some of the data has been suggested by
EPA's Beltsville, Md. Chemical Laboratory.
-------
November 1975
Fluometuron EPA-1
Determination of Fluometuron
by Infrared Spectroscopy
Fluometuron ±3 the accepted common name for l,l-dimethyl-3-
(a',Q',a'-trifluoro-ffl-tolyl) urea, a registered herbicide having the
chemical structure:
0
II
C-N
Molecular formula: C H F N.O
Molecular weight: 232.2
Melting point: 163 to 164.5°C (The technical product is about
96% pure and has a m.p. of about 155°C)
Physical state, color, and odor: odorless, white, crystalline solid
Solubility: 90 ppm in water at 25°C; soluble in acetone, ethanol,
isopropanol
Stability: stable, non-corrosive, compatible with other herbicides
Other names: Cotoran (CIBA-GEIGY), Lanex (Nor-Am), C-2059, CIBA-2059
Reagents;
1. Fluometuron standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous granular
-------
2 Fluometuron EPA-1
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.5 mm NaCl or KBr cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.1 gram fluometuron standard into a small glass-
stoppered flask or screw-cap bottle, add 50 ml chloroform by
pipette, close tightly, and shake to dissolve. Add a small
amount of anhydrous sodium sulfate to insure dryness. (final
cone 2 mg/ml)
Preparation of Sample;
For wettable powders and dusts: Weigh an amount of sample
equivalent to 0.1 gram fluometuron into a glass-stoppered flask
or screw-cap tube. Add 50 ml chloroform by pipette and 1-2 grams
anhydrous sodium sulfate. Close tightly and shake for one hour.
Allow to settle, centrifuge or filter if necessary, taking pre-
cautions to prevent evaporation. (final cone 2 mg fluometuron/ml)
For suspensions (MSMA-fluometuron suspensions containing
about 13.7% fluometuron): Weigh an amount of sample equivalent
to 0.1 gram of fluometuron (0.7 gm for 13.7% fluometuron) into a
125 ml Erlenraeyer flask that contains 5 g Na2SO . Pipette 50 ml
chloroform into the flask. Shake the sample on a mechanical shaker
for one hour. Transfer a portion of the CHC1- extract to a centri-
fuge tube and centrifuge for five minutes or until the solution is
clear. If the chloroform layer has a small insoluble layer on top,
-------
3 Fluometuron EPA-1
remove the insoluble layer with a medicine dropper. Perform the
same procedure on the standard as on the sample if this extraction
procedure is used, (cone 2 rag fluometuron/ml)
Determination:
With chloroform in the reference cell, and using the optimum
quantitative analytical settings, scan both the standard and
sample from 1410 cm to 1300 cm" (7.1 p. to 7.7 p).
Determine the absorbance of standard and sample using the
—1 —1 1
peak at 1335 cm (7.A9 p) and baseline from 1355 cm to 1300 cm
(7.38 p to 7.69 ji).
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent fluometuron as
follows:
y „ (abs. sample)(cone. std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method submitted by Mississippi State Chemical Laboratory, Box CR,
Mississippi State, Mississippi 39762.
-------
August 1975
Folpet EPA-1
Determination of Folpet
by Infrared Spectroscopy
Folpet is the acceptable common name for N-(trichloromethylthio)
phthalimide, a registered fungicide having the chemical formula:
Cl
N—S—C—Cl
Cl
Molecular formula: CgH,Cl NO S
Molecular weight: 296.6
Melting point: 177°C
Physical state and color: white crystals
Solubility: insoluble in water (1 ppm at RT); slightly soluble in
organic solvents
Stability: stable when dry; slowly hydrolyzes in water at ordinary
temperatures, rapidly at high temperatures or under
alkaline conditions; not compatible with alkaline
pesticides; non-corrosive, but decomposition products
are.
Other names: Phaltan (Chevron), Folpan, thiophal
Reagents:
1. Folpet standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
2 Folpet EPA-1
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.1 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure;
Preparation of Standard;
Weigh 0.06 gram folpet standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml chloroform by pipette, and
shake to dissolve. Add a small amount of anhydrous sodium
sulfate to insure dryness. (final cone 6 mg/ral)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.3 gram folpet into
a glass-stoppered flask or screw-cap bottle. Add 50 ml chloro-
form by pipette and 1-2 grams anhydrous sodium sulfate. Close
tightly and shake for one hour. Allow to settle; centrifuge or
filter if necessary, taking precaution to prevent evaporation.
(final cone 6 mg folpet/ml)
-------
3 Folpet EPA-1
Determination:
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1900 cm to
1650 cm'1 (5.26 p to 6.1;u).
Determine the absorbance of standard and sample using the
peak at 1755 cm (5.70 jj) and basepoint at 1850 cm (5.41 /a).
Calculation:
From the above absorbances and using the standard and
sample solution concentrations, calculate the percent of folpet
as follows:
- (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg folpet/ml chloroform gives an
absorbance of approx. 0.06 in a 0.1 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services.
-------
November 1975
Heptachlor EPA-1
Determination of Heptachlor
by Infrared Spectroscopy
Heptachlor is the accepted common name for heptachlorotetrahydro-
4,7-methanoindene (and related compounds), a registered insecticide
having the chemical structure:
Cl
Cl
H
Molecular formula: C..H-C1-
Molecular weight:
Melting point:
373.5
95 to 96°C
Physical state, color, and odor: white crystalline solid with a mild
camphor odor; the technical product contains about
72% heptachlor and 28% related compounds and is a
soft waxy solid with a melting range of 46 to 74°C
Solubility: practically insoluble in water; soluble in most organic
solvents
Stability: stable to light, moisture, air, and to moderate heat; not
readily dehydrochlorinated, but susceptible to oxidation
to heptachlor epoxide; compatible with most commonly used
insecticides and fertilizers
-------
2 Heptachlor EPA-1
Other names: Velsicol 104, E3314 (Velsicol Chem. Corp.); Drinox;
Heptamul; H-34; l,4,5,6,7,8,8-heptachloro-3a,4,7,7a-
tetrahydro-4,7-raethanoindene
Reagents:
1. Heptachlor standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Acetone, pesticide or spectro grade (dried over sodium sulfate)
4. Pentane (b.p. 20-40°C), pesticide or spectro grade
5. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.5 mm KBr or NaCl cells
2. Mechanical shaker
3. Soxhlet extraction apparatus
4. Centrifuge or filtration apparatus
5. Rotary evaporator
6. Cotton or glass wool
7. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.25 gram heptachlor standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml carbon disulfide by pipette,
and shake to dissolve. Add a small amount of anhydrous sodium
sulfate to insure dryness. (final cone 25 mg/ml)
-------
3 Heptachlor EPA-1
Preparation of Sample:
For extraction by shaking (formulations over 10%), weigh a
portion of sample equivalent to 0.5 gram heptachlor into a 125 ml
glass-stoppered or screw-cap Erlenmeyer flask, add by pipette 50 ml
of mixed solvent (9+1, carbon disulfide + dry acetone), close
tightly, and shake for 30 minutes. Allow to settle; centrifuge or
filter if necessary, taking precaution to prevent evaporation.
Pipette 25 ml into a 125 ml standard tapered flask, and evaporate
to just dryness under vacuum on a rotary evaporator. Add 5 ml
carbon disulfide and evaporate to dryness (to remove the last
traces of acetone). Dissolve in, quantitatively transfer to a
10 ml volumetric flask, and make to volume with carbon disulfide.
Add a small amount of anhydrous sodium sulfate to insure dryness.
(final cone 25 mg heptachlor/ml)
For Soxhlet extraction, weigh a portion of sample equivalent
to 0.25 gram heptachlor into a Soxhlet thimble, plug with cotton
or glass wool, and extract with pentane for two hours. Evaporate
to just dryness on a rotary evaporator. Add 5 ml carbon disulfide
and again evaporate to dryness. Dissolve in, quantitatively transfer
to a 10 ml volumetric flask, and make to volume with carbon disulfide
Add a small amount of anhydrous sodium sulfate to insure dryness.
(final cone 25 mg heptachlor/ml)
Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
_1
instrument being used, scan both the standard and sample from 700 cm
to 625 cm"1 (14.3 p to 16.0 ji) .
Determine the absorbance of standard and sample using the peak
at 658 cm (15.2 p) and baseline from 673 cm to 637 cm (14.85 jj
to 15.7 ).
-------
Heptachlor EPA-1
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent heptachlor as follows:
7 - (abs. sample)(cone, std in mg/ml)(% purity _std)
(abs. std)(cone, sample in mg/ml)
-------
November 1975
Indolebutyric acid EPA-1
Determination of Indolebutyric acid
by Ultraviolet Spectroscopy
Indolebutyric acid is 4-(3-indolyl)-butyric acid, a registered
plant growth regulator having the chemical structure:
0
CH2-CH2-CH2-C—OH
Molecular formula: C H NO
Molecular weight: 203.2
Melting point: 124°C
Physical state, color, and odor: white crystalline solid; slight
characteristic odor
Solubility: practically insoluble in water and chloroform; soluble
in alcohol, ether, acetone, and other organic solvents;
forms water-soluble alkaline salts
Stability: stable in alkaline medium
Other names: Hormodin, Seradix, 3-indolebutyric acid, indole-3-butyric acid
Reagents:
1. Indolebutyric acid standard of known % purity
2. Ethanol, pesticide or spectro grade
3. Sodium hydroxide solution, 0.5% in ethanol
-------
2 Indolebutyric acid EPA-1
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording with
matched 1 en silica cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.1 gram indolebutyric acid into a 100 ml volumetric
flask, dissolve in, and make to volume with 0.52 NaOH in ethanol
solution. Mix thoroughly and pipette 10 ml into a second 100 ml
volumetric flask. Make to volume with 0.5% NaOH in ethanol solution
and mix thoroughly. Pipette 20 ml of this second solution into a
third 100 ml volumetric flask, make to volume with water, and mix
thoroughly. (final cone 20 jug indolebutyric acid/ml and 20 ml 0.5%
NaOH in ethanol/100 ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.01 gram indolebutyric
acid into a 250 ml glass-stoppered or screw-cap Erlenmeyer flask,
add by pipette 100 ml 0.5% NaOH in ethanol solution, and shake for
3 hours. Allow to settle; centrifuge or filter if necessary,taking
precaution to prevent evaporation. Pipette 20 ml of the clear
solution into a 100 ml volumetric flask and make to volume with
water. (final cone 20 jig indolebutyric acid/ml and 20 ml 0.5% NaOH
in ethanol/100 ml)
Preparation of blank solution for reference cell:
Pipette 20 ml 0.5% NaOH in ethanol solution into a 100 ml volu-
metric flask, make to volume with water, and mix thoroughly.
-------
Indolebutyric acid EPA-1
UV jetejrjnination:
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 280 nm with the
blank solution in each cell. Scan both the standard and sample
from 360 nm to 250 nm with the blank solution in the reference
cell. Measure the absorbance of both standard and sample at
280 nm.
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent indolebutyric acid as
follows:
(abs. sample)(cone, std in pg/ml)(% purity std)
(abs. std) (cone, sample in
-------
-------
October 1975 Karbutilate EPA-1
Determination of Karbutilate in Solid
Formulations by Infrared Spectroscopy
Karbutilate is the accepted common name for m-(3,3-dimethylureido)
phenyl tert-butylcarbamate, a registered herbicide having the chemical
structure: 0 H Chta
— C — N — C— CH3
Molecular formula: C ^H^N 0
Molecular weight: 279.4
Melting point: 176 to 176.5°C
Physical state and color: white crystalline solid
Solubility: at RT — 325 ppm in water; less than 3% in isopropanol
or xylene; 20 to 25% in dimethylformamide or dimethyl
sulfoxide
Stability: stable and non-corrosive
Other names: Tandex (Niagara - FMC Corp.); NIA 11092; tert-butyl-
carbamic acid, ester with 3-(m-hydroxyphenyl)-l,l-
dimethylurea
Reagents:
1. Karbutilate standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
2 Karbutilate EPA-1
Equipment;
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.2 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, put in 1-2 grams anhydrous sodium sulfate, and
seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard;
Weigh 0.08 gram karbutilate standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml chloroform by
pipette, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 8 mg/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.4 gram karbutilate
into a glass-stoppered flask or screw-cap tube. Add 50 ml
chloroform by pipette and 1-2 grams anhydrous sodium sulfate.
Close tightly and shake for one hour. Allow to settle; centrifuge
or filter if necessary, taking precaution to prevent evaporation.
(final cone 8 rag karbutilate/ml)
-------
3 Karbutilate EPA-1
Determination;
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1925 cm to
_i
1650 cm (5.2 /i to 6.0 p) .
Determine the absorbance of standard and sample using the
peak at 1745 cm (5.73 jj) and basepoint at 1840 cm (5.43 ju) .
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent karbutilate as
follows:
7 _ (abs. sample)(cone. std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg karbutilate/ml chloroform gives an
absorbance of approx. 0.05 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services.
* based on Niagara test method #10 7/69
Beltsville Chemistry Lab, EPA, Technical Services Division, Beltsville, Md,
suggests the following:
scan range: 2000 cm" to 1600 cm" (5 ju to 6.25 ;i)
analytical peak: as above
baseline: along shoulder from about 2000 cm to 1800 cm
(5 i to 5.56 p)
-------
-------
July 1975
Determination of Linuron by
High Pressure Liquid Chromatography
Linuron EPA-1
(Tentative)
Linuron is the common name for 3-(3,4-dichlorophenyl)-l-methoxy
-1-methylurea, a registered herbicide having the chemical structure:
Molecular formula: CQHiocl2N202
Molecular weight: 249.1
Melting point: 93 to 94°C
Physical state, color, and odor: odorless, white, crystalline solid
Solubility: 75 ppm in water at 25°C; slightly soluble in aliphatic
hydrocarbons, moderately soluble in ethanol and common
aromatic solvents, soluble in acetone
Stability: stable at its m.p. and in solution; slowly decomposed by
acids and bases in moist soil; non-corrosive
Other names: Lorox (DuPont), Afalon, Sarclex, HOE 2810
Reagents:
1. Linuron standard of known % purity
2. Chloroform
3. Hexane
4. Methanol
5. Methylene chloride
All solvents should be pesticide or spectro grade.
-------
2 Linuron EPA-1
(Tentative)
Equipment;
1. High pressure liquid chromatograph
2. High pressure liquid syringe or sample injection loop
3. Liquid chromatographic column 4 mm I.D, x 25 cm packed
with LiChrosorb Si 60 - 10 ji (or equivalent column)
4. Usual laboratory glassware
Operating conditions for Hewlett-Packard Model 1010B LC;
Mobile phase: 40 ml methanol in 2000 ml of a solution containing
80% methylene chloride and 20% hexane
Column temperature: ambient
2
Observed column pressure: 3 kg/cm (425 PSI)
Flow rate: 3 ml/rain
Detector: UV at 254 nm
Chart speed: 0.5 in/min
Injection: 10 ^il
Conditions may have to be varied by the analyst for other instru-
ments, column variations, sample composition, etc. to obtain optimum
response and reproducibility.
Procedure;
Preparation of Standard:
Weigh 0.01 gram linuron standard into a 50 ml volumetric
flask; dissolve and make to volume with chloroform (final
cone 0.2 mg/ml).
-------
3 Linuron EPA-1
(Tentative)
Preparation of Sample:
Weigh an amount of sample equivalent to 0.02 gram linuron
into a 100 ml volumetric flask, make to volume with chloroform
and mix thoroughly (final cone 0.2 mg linuron/ml).
Determination:
Alternately inject three 10 jil portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent linuron as follows:
,, m (pk. ht. or area sample) (wt. std injected) (% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method developed by Joseph B. Audino, Supervisor, Pesticide Formulation
Laboratory, California Department of Food and Agriculture; and by
Yoshihiko Kawano, Associate Chemist on sabbatical leave from the
University of Hawaii.
-------
September 1975 Linuron EPA-2
Determination of Linuron
by Infrared Spectroscopy
Linuron is the common name for 3-(3,4-dichlorophenyl)-l-methoxy
-1-methylurea, a registered herbicide having the chemical structure:
CH3 Q H
CH3 0 N
Molecular formula: caHioC12N2°2
Molecular weight: 249.1
Melting point: 93 to 94°C
Physical state, color, and odor: odorless, white, crystalline solid
Solubility: 75 ppm in water at 25°C; slightly soluble in aliphatic
hydrocarbons, moderately soluble in ethanol and common
aromatic solvents, soluble in acetone
Stability: stable at its m.p. and in solution; slowly decomposed by
acids and bases in moist soil; non-corrosive
Other names: Lorox (DuPont), Afalon, Sarclex, HOE 2810
Reagents;
1. Linuron standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
2 Linuron EPA-2
Equipment:
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard;
Weigh 0.20 gram linuron standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml chloroform by pipette, and
shake to dissolve. Add a small amount of anhydrous sodium sulfate
to insure dryness. (final cone 20 mg/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 1.0 gram linuron into
a glass-stoppered flask or screw-cap tube. Add 50 ml chloroform
by pipette and 1-2 grams anhydrous sodium sulfate. Close tightly
and shake for one hour. Allow to settle; centrifuge or filter if
necessary, taking precaution to prevent evaporation, (final cone
20 mg linuron/ml)
-------
Linuron EPA-2
Pet erminat ion:
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1370 cm to
1250 cm'1 (7.3 ju to 8.0 p).
Determine the absorbance of standard and sample using the
peak at 1290 cm" (7.75 ji) and basepoint at 1258 cm" (7.95 ;u) .
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent linuron as follows:
2 « (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg linuron/ml chloroform gives an absorbance
of approx. 0.01 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services.
-------
December 1975
Malathion EPA-1
(Tentative)
Determination of Malathion by
High Pressure Liquid Chromatography
Malathion is the official common name for 0,0-dimethyl dithio-
phosphate of diethyl mercaptosuccinate, a registered insecticide
having the chemical structure:
CH3
0
II
-f~LJ f
^n~ v.
0 CH2—CH3
0
CH2—C 0—CH2—CH3
Molecular formula: C-AH._0,PS_
iu iy o /
Molecular weight: 330.A
Melting/boiling point: m.p. 2.85°C, b.p. 156 to 157°C at 0.7 mm Hg
with slight decomposition
Physical state, color, and odor: clear colorless to amber liquid,
technical grade 95% with a garlic-like odor
Solubility: 145 ppm in water; limited solubility in petroleum oils but
miscible with most organic solvents; light petroleum oil
(30-60°C) is soluble in malathion to the extent of 35%
Stability: rapidly hydrolyzed at pH above 7.0 or below 5.0 but is
stable in aqueous solutions buffered at pH 5.26; incompat-
ible with alkaline pesticides and is corrosive to iron,
hence lined containers must be used.
-------
2 Malathion EPA-1
(Tentative)
Other trames: El 4049 and Cythion (American Cyanamid), mercaptothion
(So. Africa), carbofos (USSR), Emmatos, For-Mal, Fyfanon,
Karbofos, Kop-Thion, Kypfos, Malaspray, Malamar, MLT,
Zithiol
Reagents:
1. Malathion standard of known % purity
2. Methanol, ACS
Equipment:
1. High pressure liquid chromatograph with UV detector at 254 nm.
If a variable wavelength UV detector is available, other wave-
lengths may be useful to increase sensitivity or eliminate
interference. 235 nm has been found very good for raalathion.
2. Suitable column such as:
a. DuPont ODS Permaphase, 1 meter x 2.1 mm ID
b. Perkin-Elmer ODS Sil-X II-RP, 1/2 meter x 2.6 mm ID
3. High pressure liquid syringe or sample injection loop
4. Usual laboratory glassware
Operating Conditions;
Mobile phase: 30% methanol + 70% water
Column temperature: 55°C
Chart speed: 5 min/inch or equivalent
Flow rate: 0.5 to 1.5 ml/min (Perkin-Elmer 1/2 meter column)
Pressure: 700 psi (DuPont 1 meter column)
Attenuation: Adjusted
Conditions may have to be varied by the analyst for the specific
instrument being used, column variations, sample composition, etc. to
obtain optimuK response and reproducibility.
-------
Malathlon EPA-1
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.05 gram malathion standard into a small glass-stoppered
flask or vial, add 10 ml methanol by pipette, dissolve and mix
well, (final cone 5
Preparation of Sample;
Weigh an amount of sample equivalent to 0.5 gram malathion into
a glass-stoppered flask or vial, add 100 ml metHanoi by pipette,
and shake thoroughly to dissolve the malathion. With granules or
dust, shake for 30 minutes on a mechanical shaker or shake by hand
intermittently for one hour. Allow any solid matter to settle;
filter or centrifuge if necessary, (final cone 5 ug malathion/ul)
Determination:
Using a high pressure liquid syringe or sample injection loop,
alternately inject three 5 ill portions each of standard and sample
solutions. Measure the peak height or peak area for each peak and
calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the percent
malathion as follows:
. m (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA, Beltsville, Md.
-------
December 1975 Malathion EPA-2
Determination of Malathion
by Infrared Spectroscopy
Malathion is the official common name for 0,0-dimethyl dithio-
phosphate of diethyl mercaptosuccinate, a registered insecticide
having the chemical structure:
-a ! J
P—S—CH C—0 CH2—CH3
CH3— 0'
0
II
CH2 C 0 CH2 ~ CH3
Molecular formula: C1AH. _O.PS,,
j.u iy D L
Molecular weight: 330.A
Melting/boiling point: m.p. 2.85°C, b.p. 156 to 157°C at 0.7 mm Hg
with slight decomposition
Physical state, color, and odor: clear colorless to amber liquid,
technical grade 95% with a garlic-like odor
Solubility: 145 ppm in water; limited solubility in petroleum oils but
miscible with most organic solvents; light petroleum oil
(30-60°C) is soluble in malathion to the extent of 35%
Stability: rapidly hydrolyzed at pH above 7.0 or below 5.0 but is
stable in aqueous solutions buffered at pH 5.26; incompat-
ible with alkaline pesticides and is corrosive to iron,
hence lined containers must be used.
Other names: El 4049 and Cythion (American Cyanamid), mercaptothion
(So. Africa), carbofos (USSR), Emmatos, For-Mal, Fyfanon,
Karbofos, Kop-Thion, Kypfos, Malaspray, Malamar, MLT,
Zithiol
-------
2 Malathion EPA-2
Reagents:
1. Malathion standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.2 mm KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
A. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, put in 1-2 grams anhydrous sodium sulfate, and seal
tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50 RPM
on a standard Patterson-Kelley twin shell blender that has
been modified by replacing the blending shell with a box to
hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.1 gram malathion standard into a 10 ml volumetric flask
and make to volume with carbon disulfide. Add a small amount of
anhydrous sodium sulfate to insure dryness and shake thoroughly.
(cone 10 mg/ml)
-------
Malathion EPA-2
Preparation of Sample:
For dusts, granules, and wettable powders, weigh a portion of
sample equivalent to 0.5 gram malathion into a 125 ml glass-
stoppered or screw-cap Erlenmeyer flasK. Add 50 ml carbon disul-
fide by pipette and 1-2 grams anhydrous sodium sulfate. Close
tightly, shake on a mechanical shaker for 1 hour, allow to settle and
filter or centrifuge if necessary, taking precaution to avoid
evaporation. (final cone 10 mg malathion/ml)
For emulsifiable concentrates, weigh a portion of sample
equivalent to 0.5 gram malathion into a small beaker, place on a
steam bath, and evaporate the solvent with a current of air. Add
about 5 ml of carbon disulfide and evaporate again. Extract the
cooled residue with carbon disulfide, transfer to a 50 ml volu-
metric flask, and make to volume. Add a small amount of anhydrous
sodium sulfate to insure dryness and shake thoroughly. (final
cone 10 mg malathion/ml)
An alternative procedure, especially where interfering com-
ponents cannot be removed by evaporation, is to prepare a compen-
sating solution containing approximately the same concentration
of interfering materials as is expected in the sample. This
solution is used in the reference cell of double beam instruments.
Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings, scan both the standard
and sample from 685 cm to 550 cm" (14.5 p to 18.0^0.
Determine the absorbance of standard and sample using the
peak at 657.9 cm~ (15.2 ^i) and basepoint 625 cm (16.0yu).
-------
Malathion EPA-2
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent malathion as
follows :
. sample) (cone, std in mg/ml)(% purity std)
(abs. std) (cone, sample in rag/ml)
(A concentration of 1 rag malathion/ml carbon disulfide gives an
absorbance of approx. 0.025 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
-------
November 1975
Ketaldehvde EPA-1
Determination of lletaldehyde
by lodimetric Titration
Metaldehyde is a registered molluscicide (attractant and toxicant
for slugs and snails). Chemically, it is a polymerization of acetalde-
hyde; it is thought to be a stereoisomer of the eight-membered ring:
r
0 C 0
CH-T-C—H
H—C—CH3
CH3
Molecular formula: (CK CHO)
Molecular weight:
Melting point:
(AA.l)
in sealed tube, 246°C; sublimes at 110 to 120°C
with partial depolymerization
Physical state, color, and odor: white crystalline flammable material
with a powdery appearance and mild characteristic odor
Solubility: practically insoluble in water (200 ppm at 17°C) ; low
solubility in ethanol (1.8% at 70°C) and ether; soluble
in benzene and chloroform
Stability: combustible (burns with a non-smoky flame, thus it is used
as a solid fuel); subject to depolymerization and sublima-
tion: avoid soldered tinplate containers and high
temperature
Other names: Antinilace, Meta, metacetaldehyde
-------
2 Metaldehyde EPA-1
Reagents:
1. Sulfuric acid, IN solution
2. Sodium metabisulfite, 2.5% solution - dissolve 25 grams
Na_S-0 in water and make to one liter.
^» Z. J
3. Iodine, 0.1N standard solution - dissolve 12.7 grams iodine
and 25.4 grams potassium iodide in water and make to one
liter. Standardize against an arsenic primary standard.
4. Iodine, IN solution - dissolve 63.5 grams iodine and 127
grams potassium iodide in water and make to 500 ml. (This
solution need not be standardized.)
5. Starch indicator, 1% solution - boil 1 gram soluble starch
in 100 ml water a few minutes; cool; store in bottle with
1 drop of mercury as preservative.
6. Sodium bicarbonate, powder
(All reagents should be ACS grade.)
Equipment:
1. 150 ml round-bottom distilling flask (with side arm bent
vertically downward - see below)
2. Spiral condenser fitted with a 1 mm delivery tube long enough
to reach the bottom of a 100 ml graduated cylinder
3. Thermometer 0 —100°C
4. Heating mantle or water bath for 60-70°C
5. Compressed air
6. Steam generator
7. Titration apparatus
8. Usual laboratory glassware
-------
3 Metaldehyde EPA-1
Procedure:
Apparatus assembly:
Bend the side outlet tube of a 150 ml distilling flask verti-
cally downward so that it can be attached to the top of a vertical
spiral condenser. To the bottom of the condenser, attach a
delivery tube long enough to reach just to the bottom of a 100 ml
graduated cylinder - the tip should be about 1 mm internal
diameter. The bulb of the distilling flask should be placed in
either a water bath or heating mantle so that a temperature of
60-70°C can be maintained for one hour. Fit a thermometer and an
air inlet tube through a two-hole stopper in the neck of the flask
so that both reach nearly to the bottom. The air inlet tube should
have a fitting that can be changed from compressed air to steam.
Distillation:
Weigh a portion of sample equivalent to 0.1 gram metaldehyde,
transfer to the distilling flask, add 50 ml of IN sulfuric acid
solution, and shake or swirl thoroughly so that all the sample is
wet by the acid solution. Place 40 ml of 2.5% sodium bisulfite
solution into a 100 ml graduated cylinder and place under the
condenser with the delivery tube extending almost to the bottom.
Attach the distillation flask to the assembled apparatus and heat
at 60-70°C with an air flow of about four bubbles per second.
After one hour, disconnect the air supply and immediately attach a
steam generator and distill 50 ml into the bisulfite solution.
Transfer the distillate and bisulfite solution to a 200 ml volu-
metric flask, make to volume with water, and mix well.
Titration:
Transfer 100 ml of the distillate-bisulfite solution to a 500 ml
Erlenmeyer flask, add a few ml starch indicator, titrate the excess
bisulfite solution by adding about 5 ml of the IN iodine solution,
-------
Metaldehyde EPA-1
and complete titration with 0.1N iodine solution to the exact
blue-violet endpoint. If exact endpoint is passed, add a little
bisulfite solution and re-titrate with 0.1N iodine to the exact
endpoint. Neutralize the solution with sodium bicarbonate powder
and then add 5-10 grams in excess. When the solution becomes
colorless, immediately titrate with the 0.1N iodine solution to
a blue-violet color which remains for one minute after the
addition of 1 drop of the iodine solution.
The amount of 0.1N iodine solution used between the two end-
points is used to calculate the amount of metaldehyde in the
sample.
Calculation:
(ml I2)(N I2)(0.02203)(200/100)(100)
% metaldehyde = — — -. :—.-
(grams sample)
milliequivalent weight of metaldehyde = 0.02203
1 ml 0.1000N iodine solution = 0.0022 gram metaldehyde
-------
November 1975
Metaldehyde EPA-2
(Tentative)
Determination of Metaldehyde
by Gas-Liquid Chromatography
(TCD - Internal Standard)
Metaldehyde is a registered molluscicide (attractant and toxicant
for slugs and snails). Chemically, it is a polymerization of acetalde-
hyde; it is thought to be a stereoisomer of the eight-membered ring:
0
Molecular formula: (CH.CHO)
J n
Molecular weight: (44.1)
n
Melting point: in sealed tube, 246°C: sublimes at 110 to 120°C
with partial depolymerization
Physical state, color, and odor: white crystalline flammable, material
with a powdery appearance and mild characteristic odor
Solubility: practically insoluble in water (200 ppm at 17°C); low
solubility in ethanol (1.8% at 70°C) and ether; soluble
in benzene and chloroform
Stability: combustible (burns with a non-smoky flame, thus it is used
as a solid fuel); subject to depolymcrization and sublima-
tion: avoid soldered tinplatc containers and high
temperature
Other names: Antimilace, Meta, metacetaldehyde
-------
2 Metaldehyde EPA-2
(Tentative)
Reagents:
1. Metaldehyde standard of known % purity
2. Octyl alcohol standard of known % purity
3. Chloroform, pesticide or snectro grade
4. Internal Standard solution - weigh 0.25 gram octyl alcohol
into a 100 ml volumetric flask, make to volume with chloroform,
and mix well. (cone 2.5 mg octyl alcohol/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 4' x 1/4" O.D. glass column packed with 3% XE-60
on Chromosorb G AW DMCS (or equivalent column)
3. Precision liquid syringe: 10 ul
4. Usual laboratory glassware
Operat±n^_Condit±ons_ for TCP:
Column temperature: 90°C
Injection temperature: 140°C
Detector temperature: 140°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas flow rate: 30 ml/rain
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response and
reproducibility.
-------
3 Metaldehyde EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.14 gram metaldehyde standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 10 ml of the internal
standard solution and shake to dissolve. (final cone 14 mg metalde-
hyde and 2.5 mg octyl alcohol/ml)
.^r.e£-a£a-L:'-2.n °JL _§aiT!P!.e_:
Weigh a portion of sample eauivalent to 0.35 gram metaldehyde
into a small glass-stoppered flask or screw-can bottle. Add by
pipette 25 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the metaldehyde.
For coarse or granular materials, shake mechanically for 30
minutes or shake by hand intermittently for one hour. (final cone
14 mg metaldehyde and 2.5 mg octyl alcohol/ml)
Determination:
Inject 4-6 til of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is octyl alcohol, then
metaldehyde.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of metaldehyde and octyl
alcohol from both the standard-internal standard solution and the
sample-internal standard solution.
-------
Metaldehvde
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
ity octyl alcohol) (pk. ht. or area metaldehyde)
- -- ----
(wt . metaldehyde) (% purity metaldehyde) (pk. ht . or area octyl alcohol)
Determine the percent metaldehyde for each injection of the
sample-internal standard solution as follows and calculate the
average:
_ (wt . octyl alcohol) (% purity octyl alcohol) (pk. ht. or area metaldehyde) QQQ±
(wt. sample) (pk. ht. or area octyl alcohol) (RF) \fjt~0
Method submitted by Stelios Gerazounis, EPA, Region II, New York, K.Y.
Some additional information and a few changes have been made in this
write-up; therefore, any comments, criticisms, suggestions, data, etc.
concerning this method will be appreciated.
-------
November 1975
Metaldehyde EPA-3
(Tentative)
Determination of Metaldehyde
by Infrared Spectroscopy
Metaldehyde is a registered molluscicide (attractant and toxicant
for slugs and snails). Chemically, it is a polymerization of acetalde-
hyde; it is thought to be a stereoisomer of the eight-membered ring:
0
0
Molecular formula: (CH_CHO)
3 n
Molecular weight:
Melting point:
(44.1)
n
in sealed tube, 246°C; sublimes at 110 to 120°C
with partial depolymerization
Physical state, color, and odor: white crystalline flammable material
with a powdery appearance and mild characteristic odor
Solubility: practically insoluble in water (200 ppm at 17°C) ; low
solubility in ethanol (1.8% at 70°C) and ether; soluble
in benzene and chloroform
Stability: combustible (burns with a non-smoky flame, thus it is used
as a solid fuel); subject to depolymerization and sublima-
tion: avoid soldered tinplate containers and high
temperature
Other names: Antimilace, Meta, metacetaldehyde
-------
2 Metaldehyde EPA-3
(Tentative)
Reagents:
1. Metaldehyde standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.2 mm NaCl or KBr cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.06 gram metaldehyde standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml chloroform by pipette, close
tightly, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 6 mg/ml)
Preparat ion_ of Samp le_:
Weigh an amount of sample equivalent to 0.3 gram metaldehyde
into a glass-stoppered flask or screw-cap tube. Add 50 ml chloroform
by pipette and 1-2 grams anhydrous sodium sulfate. Close tightly and
shake for one hour. Allow to settle; centrifuge or filter if necessary,
taking precautions to prevent evaporation. (final cone 6 mg metalde-
hyde /ml)
Dejtejrm inatjLoii:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings, scan both the standard and
sample from 1250 cm~ to 1110 cm" (8.0 ji to 9.0ju).
-------
Metaldehyde EPA-3
(Tentative)
Determine the absorbance of standard and sample using the peak
at 1164 cm'1 (8.59 u) and basepoint 1140 cm~ (8.77yj).
Caleula tion:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent metaldehyde as
follows:
7 a. (abs. sample)(cone.std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method submitted by E. Greer, EPA, Region IX, San Francisco, California.
-------
November 1975
Metaldehyde EPA-4
(Tentative)
Determination of Metaldehyde
by Gas-Liquid Chromatography (TCD)
Metaldehyde is a registered molluscicide (attractant and toxicant
for slugs and snails). Chemically, it is a polymerization of acetalde
hyde; it is thought to be a stereoisomer of the eight-membered ring:
0
CH3
— c o
H
CH3—C—H H—C
V
0 C 0
-CH3
Molecular formula: (CH.CHO)
n
Molecular weight:
Melting point:
(44.1)
n
in sealed tube, 246°C; sublimes at 110 to 120°C
with partial depolymerization
Physical state, color, and odor: white crystalline flammable material
with a powdery appearance and mild characteristic odor
Solubility: practically insoluble in water (200 ppm at 17°C) ; low
solubility in ethanol (1.8% at 70°C) and ether; soluble
in benzene and chloroform
Stability: combustible (burns with a non-smoky flame, thus it is used
as a solid fuel); subject to depolymerization and sublima-
tion: avoid soldered tinplate containers and high
temperature
Other names: Antimilace, Meta, metacetaldehyde
-------
2 Metaldehyde EPA-4
(Tentative)
Reagents:
1. Metaldehyde standard of known % purity
2. Chloroform, pesticide or spectro grade
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 5' x 1/4" O.D. glass column packed with 20% SE-30 on
Chromosorb W AW DMCS (or equivalent column)
3. Precision liquid syringe: 50 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for TCD:
Column temperature: 120°C
Injection temperature: 160°C
Detector temperature: 160°C
Carrier gas: Helium
Flow rate: 30 ml/min
Operating conditions for filament current, column temperature, or
gas flow should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.06 gram metaldehyde standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml chloroform by
pipette, and shake to dissolve. (final cone 6 mg/ml)
-------
Metaldehyde EPA-4
(Tentative)
Weigh a portion of sample equivalent to 0.3 gram metaldehyde
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 50 ml chloroform, close tightly, and shake thoroughly to
dissolve and extract the metaldehyde. For coarse or granular
materials, shake mechanically for 30 minutes or shake by hand
intermittently for one hour. (final cone 6 mg metaldehyde/ml)
Using a precision liquid syringe, alternately inject three
5-10 ul portions each of standard and sample solutions. Measure
the peak height or peak area for each peak and calculate the
average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent metaldehyde as follows:
7 - (pk- ht. or area sample) (w^t^. std injected) (% purity of std)
(pk. ht . or area standard) (wt. sample injected)
Method submitted by Eva Santos, EPA Region IX, San Francisco, California.
-------
December 1975 Methiocarb EPA-1
(Tentative)
Determination of Methiocarb in Solid Formulations
by Infrared Spectroscopy
Methiocarb is a common name (BSI) for 4-(methylthio)-3,5-xylyl
N-methylcarbamate, a registered insecticide and acaricide having the
chemical structure:
Molecular formula:
Molecular weight:
Melting point:
CH-i
\
-ff ^
CH3
CUH15H02S
225.3
121'C
\ 0
\ II
\\ n r
/ C
H
N—CH
Physical state, color, and odor: white crystalline powder with a mild
milk-like odor
Solubility: insoluble in water; soluble in acetone and alcohol;
soluble in most organic solvents
Stability: unstable in highly alkaline media (hydrolyzed by alkalis)
Other names: Mesurol, Bay 37344, H 321, (Bayer AG); mercaptodimethur,
metmercapturon, Draza
Reagents;
1. Methiocarb standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
Methiocarb EPA-1
(Tentative)
Equipment:
1. Infrared spectrophotometer, double beam ratio recording, with
matched 0.2 mm NaCl or KBr cells
2. Mechanical shaker
3. Soxhlet extraction apparatus
4. Cotton or glass wool
5. Centrifuge or filtration apparatus
6. Rotary evaporator
7. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.07 gram methiocarb standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml chloroform by
pipette, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 7 rag/ml)
Preparation of Sample:
For high percent formulations (more than 10%), weigh a portion
of sample equivalent to 0.35 gram methiocarb into a glass-stoppered
flask or screw-cap bottle. Add 50 ml chloroform by pipette and 1-2
grams anhydrous sodium sulfate. Close tightly and shake for one
hour. Allow to settle; centrifuge or filter if necessary,taking
precaution to prevent evaporation, (final cone 7 mg methiocarb/ml)
For low percent (less than 10%) formulations, weigh a portion
of sample equivalent to 0.35 gram methiocarb into a Soxhlet extrac-
tion thimble, plug with cotton or glass wool, and extract with
chloroform for three hours. Evaporate to about 25 ml on a rotary
evaporator, quantitatively transfer to a 50 ml volumetric flask,
?•
and make to volume with chloroform. Add a small amount of anhydrous
sodium sulfate to clarify and dry the solution, (final cone 7 mg
methiocarb/ml)
-------
Methiocarb EPA-1
(Tentative)
IR Determination;
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1880 cm to
1625 cm"1 (5.32 ji to 6.15 ji).
Determine the absorbance of standard and sample using the
peak at 1748 cm" (5.72 ji) and a baseline from 1835 cm"1 to
1667 cm"1 (5.45 u to 6.00 ji) .
Calculation:
From the above absorbances, calculate the percent methiocarb
as follows:
„ m (abs. sample)(cone. std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
-------
September 1975
Methoxychlor EPA-1
(Tentative)
Determination of Technical Methoxychlor
by Infrared Spectroscopy
Methoxychlor, technical is the official name for 2,2-bis
(p-methoxyphenyl)-l,l,l-trichloroethane 88% and related compounds
12%; it is a registered insecticide having the chemical structure:
Cl C—Cl
Cl
Molecular formula: C,,H,CC1_00
lO 13 J L
Molecular weight: 345.5
Physical state, color, and odor: pure p,p* isomer forms colorless crystals;
technical product is a gray flaky powder containing
88% p,p* isomer with the bulk of the remainder being
the o,p isomer
Melting point: pure p,p' isomer 89°C; technical 70 to 85°C
Solubility: practically insoluble in water; moderately soluble in
ethanol and petroleum oils; readily soluble in most aromatic
solvents
Stability: resistant to heat and oxidation; susceptible to dehydro-
chlorination by alcoholic alkali and heavy metal catalyst
Other names: Marlate (DuPont), Moxie, l,l,l-trichloro-2,2-bis(p-methoxy-
phenyl)ethane
-------
2 Methoxychlor EPA-1
(Tentative)
Reagents;
1. Methoxychlor, technical standard (minimum 88% p,p' isomer)
2. Carbon disulfide, pesticide or spectre grade
3. Sodium sulfate, anhydrous, granular
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm KBr or NaCl cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure;
Preparation of Standard:
Weigh 0.16 gram technical methoxychlor standard into a small
glass-stoppered flask or screw-cap bottle, add 10 ml carbon
disulfide by pipette, close tightly, and shake to dissolve. Add
a small amount of anhydrous sodium sulfate to insure dryness.
(final cone 16 mg/ml)
Preparation of Sample;
Weigh an amount of sample (dusts and wettable powders) equiv-
alent to 1.6 grams technical methoxychlor into a glass-stoppered
flask or screw-cap bottle. Add 100 ml carbon disulfide by pipette
and 1-2 grams anhydrous sodium sulfate. Close tightly and shake
for one hour. Allow to settle; centrifuge or filter if necessary,
taking precaution to prevent evaporation, (final cone 16 mg tech.
me thoxychlor/ml)
-------
3 Methoxychlor EPA-1
(Tentative)
(Aerosols, emulsiftable concentrates, and oil solutions may
be tried by this method; however, interfering substances are
most likely to be present.)
Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical setting for the particular 1R
instrument being used, scan both the standard and sample from
870 cm'1 to 740 cm"1 (11.5 p to 13.5 ;i).*
Determine the absorbance of standard and sample using the
peak at 795.5 cm (12.57 ji) and basepoint at 772.2 cm (12.95
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent technical methoxy-
chlor as follows:
2 m (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method contributed by M. Conti and N. Frost, EPA Region IX, San Francisco,
California.
* Absorption bands at 1250 cm~ (8.0 ji), 1179 cm (8.48 /i) , 1042 cm~
(9.6 ju), or 752 cm (13.3 p) may also be used when interference from
other ingredients is present; however, the linearity should be checked.
-------
November 1975
Methoxychlor EPA-2
(Tentative)
Determination of Methoxychlor
by Gas—Liquid Chromatography
(FID - Internal Standard)
Methoxychlor, technical is the official name for 2,2-bis
(p~methoxyphenyl)-l,l,l-trichloroethane 88% and related compounds 12%;
it is a registered insecticide having the chemical structure:
CH3—0 ('
0—CH3
Molecular formula: C,,H, ,C1,.CL
lo Ij 3 i
Molecular weight: 345.5
Physical state, color, and odor: pure p,p' isomer forms colorless crystals;
technical product is a gray flaky powder containing
88% p,p' isomer with the bulk of the remainder being
the o,p isomer
Melting point:
pure p,p' isomer 89°C; technical 70 to 85°C
Solubility: practically insoluble in water; moderately soluble in ethanol
and petroleum oils; readily soluble in most aromatic solvents
Stability: resistant to heat and oxidation; susceptible to dehydrochlor-
ination by alcoholic alkali and heavy metal catalyst
Other names: Marlate (DuPont), Moxie, l,l,l-trichloro-2,2-bis(p-methoxy-
phenyl) ethane
-------
2 Methoxychlor EPA-2
(Tentative)
Reagents:
1. Methoxychlor standard of known % purity
2. Dieldrin standard of known HEOD content
3. Acetone, pesticide or spectro grade
A. Internal Standard solution - weigh 0.2 gram HEOD into a 50 ml
volumetric flask; dissolve in and make to volume with acetone.
(cone A mg HEOD/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: A' x 2 mm ID glass column packed with 5% OV-210 on
80/100 mesh Chromosorb W HP (or equivalent column)
3. Precision liquid syringe: 5 or 10 fil
A. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 190°C
Injection temperature: 2AO°C
Detector temperature: 2AO°C
Carrier gas: Nitrogen
Carrier gas pressure: AO-60 psi
Hydrogen pressure: 20 psi
Air pressure: 30 psi
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Methoxychlor EPA-2
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.1 gram methoxychlor standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 20 ml of the
internal standard solution and shake to dissolve, (final cone
5 mg methoxychlor and A mg HEOD/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.1 gram methoxychlor
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the methoxychlor.
For coarse or granular materials, shake mechanically for 30
minutes or shake by hand intermittently for one hour, (final
cone 5 mg methoxychlor and 4 mg HEOD/ml)
Determination:
Inject 1-2 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is HEOD, then methoxychlor.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of methoxychlor and HEOD
from both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
-------
RF
Methoxychlor EPA-2
(wt. HEOD)(% purity HEOD)(pk. ht. or area methoxychlorX
(wt. methoxychlor)(% purity methoxychlor)(pk. ht. or area HEOD)
Determine the percent methoxychlor for each injection of the
sample-internal standard solution as follows and calculate the
average:
% » (wt. HEOD)(% purity HEOD)(pk. ht. or area methoxychlor)
(wt. sample)(pk. ht. or area HEOD)(RF) f
Note! MG-264 interferes with dieldrin under these conditions.
This method was submitted by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond, Virginia
23219.
This method has been designated as tentative since it is a Va.
Exp. method and because some of the data has been suggested by
EPA's Beltsville Chemistry Lab. Any comments, criticisms,
suggestions, data, etc. concerning this method will be appreciated,
-------
November 1975
Methyl Parathion EPA-1
(Tentative)
Determination of Methyl Parathion
by High Pressure Liquid Chroraatography
Methyl parathion is the common (US) name (parathion-methyl, ISO
and BSI) for 0,0-dimethyl 0-p-nitrophenyl phosphorothioate, a regis-
tered insecticide having the chemical structure:
CH
N02
Molecular formula: C H NO PS
o 1U j
Molecular weight:
Melting point:
263.2
35-36°C
Physical state and color: white crystalline solid; the technical
product is a light to dark tan liquid of about
80% purity, crystallizing at about 29°C.
Solubility: 55-60 ppra in water at 25°C; slightly soluble in light
petroleum and mineral oils; soluble in most other
organic solvents
Stability: hydrolyzed by alkalis; compatible with most non-alkaline
pesticides; isomerizes on heating; it is a good methylating
agent.
Other names: Dalf (Bayer); Metacide, Nitrox 80 (Chemagro); parathion-
methyl (ISO and BSI); Metaphos (USSR); dimethyl parathion;
E601; Folidol M; Fosferno M50; Gearphos; Metron; Partron M;
Tekwaisa; Wofatox
-------
2 Methyl Parathion EPA-1
(Tentative)
Reagents:
1. Methyl parathion standard of known % purity
2. Methanol, pesticide or spectro grade
Equipment;
1. High pressure liquid chrotnatograph with UV detector at 254 nm.
If a variable wavelength UV detector is available, other
wavelengths may be useful to increase sensitivity or eliminate
interference.
2. Suitable column such as:
a. DuPont ODS Permaphase, 1 meter x 2.1 mm ID
b. Perkin-Elmer ODS Sil-X 11 RP, 1/2 meter x 2.6 mm ID
3. High pressure liquid syringe or sample injection loop
4. Usual laboratory glassware
Operating Conditions:
Mobile phase: 20% methanol + 80% water
Column temperature: 50-55°C
Chart speed: 5 mln/inch or equivalent
Flow rate: 0.5 to 1.5 ml/min (Perkin—Elmer 1/2 meter column)
Pressure: 700 psi (DuPont 1 meter column)
Attenuation: Adjusted
Conditions may have to be varied by the analyst for the specific
instrument being used, column variations, sample composition, etc.
to obtain optimum response and reproducibility.
-------
Methyl Parathion EPA-1
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.1 gram methyl parathion standard into a small glass-
stoppered flask or vial, add 100 ml methanol by pipette, dissolve
and mix well, (final cone 1 mg methyl parathion/ml)
Preparation of Sample:
Weigh an amount of sample equivalent to 0.1 gram methyl
parathion into a glass-stoppered flask or vial, add 100 ml
methanol by pipette,and shake thoroughly to dissolve the methyl
parathion. Allow any solid matter to settle; filter or centrifuge
if necessary. (final cone 1 mg methyl parathion/ml)
Determination:
Alternately inject three 10 ul portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the percent
methyl parathion as follows:
« m (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA, Beltsville, Md.
-------
November 1975 Methyl Parathion EPA-2
Determination of Methyl Parathion
by Infrared Spectroscopy
Methyl parathion is the common (US) name (parathion-methyl, ISO
and BSI) for 0,0-dimethyl Op-nitrophenyl phosphorothioate, a regis-
tered insecticide having the chemical structure:
Molecular formula: C0H,-NO-PS
o 1U _>
Molecular weight: 263.2
Melting point: 35-36°C
Physical state and color: white crystalline solid; the technical
product is a light to dark tan liquid of about
80% purity, crystallizing at about 29°C.
Solubility: 55-60 ppm in water at 25°C; slightly soluble in light
petroleum and mineral oils; soluble in most other
organic solvents
Stability: hydrolyzed by alkalis; compatible with most non-alkaline
pesticides; isomerizes on heating; it is a good methylating
agent.
Other names: Dalf (Bayer); Metacide, Nitrox 80 (Chemagro); parathion-
methyl (ISO and BSI); Metaphos (USSR); dimethyl parathion;
E601; Folidol M; Fosferno M50; Gearphos; Metron; Partron M;
Tekwaisa; Wofatox
-------
2 Methyl Parathion EPA-2
Reagents; ^^
1. Methyl parathion standard of known % purity
2. Acetone, pesticide or spectro grade
3. Carbon disulfide, pesticide or spectro grade
4. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm NaCl or KBr cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.1 gram methyl parathion into a small glass-stoppered
flask or screw-cap bottle, add 10 ml carbon disulfide by pipette,
and shake to dissolve. Add a small amount of anhydrous sodium
sulfate to insure dryness. (final cone 10 mg methyl parathion/ml)
Preparation of Sample;
For emulsiftable concentrates, weigh a portion of sample
equivalent to 0.1 gram methyl parathion into a. 10 ml volumetric
flask, make to volume with carbon disulfide, and mix well. Add a
small amount of anhydrous sodium sulfate to insure dryness. (final
cone 10 mg methyl parathion/ml)
-------
3 Methyl Parathion EPA-2
For granular formulations, weigh a portion of sample equivalent
to 0.2 gram methyl parathion into a glass-stoppered flask or screw-
cap bottle. Add 100 ml acetone by pipette and 1-2 grams anhydrous
sulfate. Close tightly and shake for one hour. Allow to settle;
centrifuge or filter if necessary, taking precaution to prevent
evaporation. Evaporate a 50 ml aliquot to dryness on a water
bath using a gentle stream of dry air; evaporate the last one or
two ml with air only. Add 5 ml carbon disulfide and evaporate
again to remove all traces of acetone. Dissolve in about 4-5 ml
carbon disulfide, transfer to a 10 ml volumetric flask, and make
to volume with carbon disulfide. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 10 mg methyl para-
thion/ml)
Determination :
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan the standard and sample from 1350 cm
to 1110 cm"1 (7.4 to 9.0 i).
Determine the absorbance of standard and sample using the peak
at 1234.6 cm" (8.10 u) and baseline from 1274 cm to 1198 cm"1
(7.85 u to 8.35 u) .
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent methyl parathion as follows:
„ ^ (abs. sample) (cone . std in mg/ml)(% purity std)
(abs. std) (cone, sample in ing /ml)
-------
November 1975
Methyl Parathion EPA-3
Determination of Methyl Parathion
in Dusts and Wettable Powder
by Colorimetric (Visible) Spectroscopy
Methyl parathion is the common (US) name (parathion-tnethyl, ISO
and BSI) for 0,0-ditnethyl 0-p-nitrophenyl phosphorothioate, a regis-
tered insecticide having the chemical structure:
Molecular formula: CQHinNO_PS
o 1U j
Molecular weight: 263.2
Melting point: 35-36°C
Physical state and color: white crystalline solid; the technical
product is a light to dark tan liquid of about
80% purity, crystallizing at about 29°C.
Solubility: 55-60 ppm in water at 25°C; slightly soluble in light
petroleum and mineral oils; soluble in most other
organic solvents
Stability: hydrolyzed by alkalis; compatible with most non-alkaline
pesticides; isomerizes on heating; it is a good methylating
agent.
Other names: Dalf (Bayer); Metacide, Nitrox 80 (Chemagro); parathion-
methyl (ISO and BSI); Metaphos (USSR); dimethyl parathion;
E601; Folidol M; Fosferno M50; Gearphos; Metron; Partron M;
Tekwaisa; Uofatox
-------
2 Methyl Parathion EPA-3
Principle of the Method:
The methyl parathion is extracted with alcohol and hydrolyzed with
potassium hydroxide in the presence of hydrogen peroxide (this prevents
reduction of the nitro group) to potassium p-nitrophenate, which is
determined colorimetrically. Any free p-nitrophenol present is deter-
mined on a portion of the extract before hydrolysis. A high free
p-nitrophenol content may indicate product decomposition,especially if
the methyl parathion assay is low.
Reagents:
1. p-Nitrophenol of known % purity
2. 95% Ethanol, ACS
3. Ethanol, 50% in water
4. Potassium hydroxide, IN solution in ethanol
5. Hydrogen peroxide, 30%
Equipment;
1. UV-VIS spectrophotometer, double beam ratio recording with
matched 1 cm cells (a photoelectric colorimeter with a filter
giving maximum transmission between 400-450 nm may be used)
2. Reflux apparatus
3. Usual laboratory glassware
Procedure;
Preparation of Standard:
Weigh 0.06 gram p-nitrophenol into a 100 ml volumetric flask;
dissolve and make to volume with 95% ethanol. Pipette 10 ml into
a second 100 ml volumetric flask and make to volume with 95%
-------
3 Methyl Parathion EPA-3
ethanol. Pipette 5 ml into a third 100 ml volumetric flask, add
by pipette 5 ml IN potassium hydroxide solution, and make to
volume with 95% ethanol. The final concentration will be 3 ug/ml.
preparation of Sample;
Weigh a portion of sample equivalent to 0.012 gram methyl
parathion into a 250 ml glass-stoppered flask. Add by pipette
100 ml 95% ethanol and shake periodically for about ten minutes.
Filter 25-50 ml into a glass-stoppered flask or bottle. If
necessary, extract a larger sample and aliquot, using 95%
ethanol as the solvent.
Determination:
Standard:
With the UV-VIS spectrophotometer at the optimum quanti-
tative settings, balance the pen for 0 and 100% at 405 nm
with 50% ethanol in both cells. Set the instrument to scan
from 500 nm to 350 nm. Scan the standard p-nitrophenol
solution between these wavelengths using 50% ethanol in the
reference cell.
Free p-nitrophenol:
To measure the free p-nitrophenol, pipette 10 ml of the
filtered sample solution into a 100 ml volumetric flask and
make to volume with 50% ethanol. Add 5 drops of IN potassium
hydroxide solution, mix, and immediately (within 2 minutes of
adding the alkali) scan from 500 nm to 350 nm. This is the
absorbance due to the free p-nitrophenol in the sample.
Methyl parathion (as p-nitrophenol):
To determine the methyl parathion (as p-nitrophenol),
pipette 5 ml of the filtered sample solution into a 125 ml
standard taper Erlenmeyer flask. Add 5 ml IN potassium
-------
4 Methyl Parathion EPA-3
hydroxide solution by pipette, 2 ml of 30% hydrogen peroxide,
a few glass beads, and reflux for at least 30 minutes. Cool,
transfer to a 100 ml volumetric flask with 50% ethanol, and
make to volume with the 50% ethanol. Scan between 500 nm
and 350 nm. This is the uncorrected total absorbance due to
the free p-nitrophenol and to the p-nitrophenol from the
methyl parathion. The concentration of this solution is 6 tig
methyl parathion/ml or approx. 3 ^ig p-nitrophenol/ml.
Calculation:
Using the absorbance due to the free p-nitrophenol (FPNP) ,
calculate the percent present as follows:
j « (abs. FPNP)(wt. std) (1/100) (10/100) (5/100) (100)
* (abs. std)(wt. sample) (1/100) (10/100)
Using the absorbance from the uncorrected total p-nitrophenol
(UTPNP) , calculate the percent as follows :
„ (abs. UTPNP) (wt. std) (1/100) (10/100) (5/100) (100)
(abs. std)(wt. sample) (1/100) (5/100)
The percent p-nitrophenol due to the methyl parathion is found
by subtracting the free p-nitrophenol from the uncorrected total
p-nitrophenol .
% p-nitrophenol » % uncorrected total p-nitrophenol - Z free p-nitrophenol
The % methyl parathion is then found by dividing this %
p-nitrophenol by .5285 or multiplying by 1.892.
% Methyl Parathion = ""1*"0 °r (1-892><% p-nitrophenol)
Methyl parathion - 52.85% p-nitrophenol'
% methyl parathion - % p-nitrophenol X 1.892
-------
November 1975
Methyl Parathion EPA-4
Determination of Methyl Parathion in Emulsifiable
Concentrates by Gas-Liquid Chromatography
(FID - Internal Standard)
Methyl parathion is the common (US) name (parathion-methyl, ISO
and BSI) for 0,0-dimethyl 0-p-nitrophenyl phosphorothioate, a regis-
tered insecticide having the chemical structure:
Molecular formula: C0H.-NOCPS
o 1U j
Molecular weight: 263.2
Melting point: 35-36°C
Physical state and color: white crystalline solid; the technical
product is a light to dark tan liquid of about
80% purity, crystallizing at about 29°C.
Solubility: 55-60 ppm in water at 25°C; slightly soluble in light
petroleum and mineral oils; soluble in most other
organic solvents
Stability: hydrolyzed by alkalis; compatible with most non-alkaline
pesticides; isomerizes on heating; it is a good methylating
agent.
Other names: Dalf (Bayer); Metacide, Nitrox 80 (Chemagro); parathion-
methyl (ISO and BSI); Metaphos (USSR); dimethyl parathion;
E601; Folidol M; Fosferno M50; Gearphos; Metron; Partron M;
Tekwaisa; Wofatox
-------
2 Methyl Parathion EPA-4
Reagents:
1. Methyl parathion standard of known % purity
2. p,p'-DDE standard of known % purity
3. Carbon disulfide, pesticide or spectro grade
4. Internal Standard solution - weigh 0.125 gram p,p'-DDE into a
25 ml volumetric flask, dissolve in, and make to volume with
carbon disulfide. (cone 5 mg DDE/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 61 x 1/4" OD glass column packed with a 1:1 mixture
of 10% DC-200 and 15% QF-1 on 80/100 mesh Gas Chrom Q
(or equivalent column)
3. Precision liquid syringe: 10 or 50 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 190°C
Injection temperature: 215°C
Detector temperature: 260°C
Carrier gas: Nitrogen
Carrier gas flow rate: 90 ml/min
Hydrogen flow rate: Adjust for specific GC
Air flow rate: Adjust for specific GC
-------
3 Methyl Parathion EPA-4
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and
reproducibility.
Procedure:
Preparation of Standard;
Weigh 0.08 gram methyl parathion standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 10 ml of the
internal standard solution and shake to dissolve, (final cone
8 mg methyl parathion and 5 mg DDE/ml)
Preparation of Sampler
Weigh a portion of sample equivalent to 0.08 gram methyl
parathion into a small glass-stoppered flask or screw-cap bottle.
Add by pipette 10 ml of the internal standard solution. Close
tightly and shake thoroughly to dissolve and extract the methyl
parathion. For coarse or granular materials, shake mechanically
for 30 minutes or shake by hand intermittently for one hour.
(final cone 8 mg methyl parathion and 5 mg DDE/ml)
Determination:
Inject 2-3 jil of standard and adjust the instrument parameters
and the volume injected to give a complete separation within a
reasonable time and peak heights of from 1/2 to 3/4 full scale.
The elution order is methyl parathion, then DDE.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
-------
Methyl Parathion EPA-4
Calculation:
Measure the peak heights or areas of methyl parathion and DDE
from both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. DDE)(% purity DDE)(pk. ht. or area methyl parathion)
(wt. methyl parathion)(% purity methyl parathion)(pk. ht. or area DDE)
Determine the percent methyl parathion for each injection of
the sample-internal standard solution as follows and calculate the
average:
. (wt. DDE)(% purity DDE)(pk. ht. or area methyl parathion) (J=
(wt. sample)(pk. ht. or area DDE)(RF)
Method submitted by Mississippi State Chemical Laboratory, Box CR,
Mississippi State, Mississippi 39762.
-------
November 1975
Methyl Parathion EPA-5
Determination of Methyl Parathion
by Gas-Liquid Chromatography
(FID - Internal Standard)
Methyl parathion is the common (US) name (parathion-methyl, ISO
and BSI) for 0, Odimethyl O-p-nitrophenyl phosphorothioate, a regis-
tered insecticide having the chemical structure:
N02
Molecular formula: CQHir.NOcPS
o 1U D
Molecular weight: 263.2
Melting point: 35-36°C
Physical state and color: white crystalline solid; the technical
product is a light to dark tan liquid of about
80% purity, crystallizing at about 29°C.
Solubility: 55-60 ppm in water at 25°C; slightly soluble in light
petroleum and mineral oils; soluble in most other
organic solvents
Stability: hydrolyzed by alkalis; compatible with most non-alkaline
pesticides; isomerizes on heating; it is a good methylating
agent.
Other names: Dalf (Bayer); Metacide, Nitrox 80 (Chemagro); parathion-
methyl (ISO and BSI); Metaphos (USSR); dimethyl parathion;
E601; Folidol M; Fosferno M50; Gearphos; Metron; Partron M;
Tekwaisa; Wofatox
-------
2 Methyl Parathion EPA-5
Reagents;
1. Methyl parathion standard of known % purity
2. Dieldrin standard of known HEOD content
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.15 gram HEOD into a
25 ml volumetric flask; dissolve in and make to volume with
acetone, (cone 6 mg HEOD/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 4 mm ID glass column packed with 3% OV-1 on
60/80 mesh Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 pi
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 175°C
Injection temperature: 250°C
Detector temperature: 250°C
Carrier gas: Nitrogen
Carrier gas pressure: (not stated in method)(40-60 psi)
Hydrogen pressure: 32 psi
Air pressure: 29 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response and
reproducibility.
-------
3 Methyl Parathion EPA-5
Procedure:
Preparation of Standard:
Weigh 0.06 gram methyl parathion standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 10 ml of the
internal standard solution and shake to dissolve, (final cone
6 rag methyl parathion and 6 mg HEOD/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.06 gram methyl
parathion into a small glass-stoppered flask or screw-cap bottle.
Add by pipette 10 ml of the internal standard solution. Close
tightly and shake thoroughly to dissolve and extract the methyl
parathion. For coarse or granular materials, shake mechanically
for 30 minutes or shake by hand intermittently for one hour.
(final cone 6 mg methyl parathion and 6 mg HEOD/ml)
Determination;
Inject 1-2 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is methyl parathion, then
HEOD.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
(Calculation:
Measure the peak heights or areas of methyl parathion and HEOD
from both the standard-internal standard solution and the sample-
internal standard solution.
-------
Methyl Parathion EPA-5
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
= (wt. HEOD)(% purity HEOD)(pk. ht. or area methyl parathion)
(wt. methyl parathion)(% purity methyl parathion)(pk. ht. or area HEOD)
Determine the percent methyl parathion for each injection of
the sample-internal standard solution as follows and calculate the
average:
7 « (wt. HEOD)(% purity HEOD) (pk. ht. or area methyl parathion)-Q^)"
= (wt. sample) (pk. ht. or area HEOD) (RF) /6/-/J
The above method was submitted by Division of Regulatory Services,
Kentucky Agricultural Experiment Station, University of Kentucky,
Lexington, Kentucky 40506.
A similar method (data below) was submitted by the Commonwealth of
Virginia, Division of Consolidated Laboratory Services, 1 North 14th Street,
Richmond, Virginia 23219.
Column: 4' x 2 mm ID glass packed with 5% SE-30 on 80/100 mesh
Chromosorb W HP
Column temp: 180C
Internal standard: Alachlor 2 mg/ml
Methyl parathion cone: 2 mg/ml
Comments, criticisms, suggestions, data, etc. concerning this method
are invited and are welcome.
-------
September 1975
Metobromuron EPA-1
(Tentative)
Determination of Metobromuron
by Infrared Spectroscopy
Metobromuron is the accepted common name for 3-(p-bromophenyl)-l-
methoxy-1-methylurea, a registered herbicide having the chemical
structure:
0-CH3
Molecular formula: C H BrN 0
Molecular weight: 259
Melting point: 95.5 to 96°C
Physical state and color: white crystalline solid
Solubility: 330 ppm in water at RT; very soluble in acetone,
chloroform, ethanol
Stability: stable; non-corrosive; good compatibility
Other names: Patoran (CIBA), C-3126
Reagents;
1. Metobromuron standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
Metobromuron EPA-1
(Tentative)
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.5 mm KBr or NaCl cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard;
Weigh 0.1 gram metobromuron standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml chloroform by
pipette, close tightly, and shake to dissolve. Add a small
amount of anhydrous sodium sulfate to insure dryness. (final
cone 10 mg/ml)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.5 gram metobromuron
into a glass-stoppered flask or screw-cap bottle. Add 50 ml
chloroform by pipette and 1-2 grams anhydrous sodium sulfate.
Close tightly and shake for one hour. Allow to settle; centri-
fuge or filter if necessary, taking precautions to prevent
evaporation, (final cone 10 metobromuron/ml)
Determination;
With chloroform in the reference cell, and using the optimum
quantitative analytical settings, scan both the standard and
-1 -1
sample from 1430 cm to 1250 cm (7.0 ji to S.O^i).
Determine the absorbance of standard and sample using the
peak at 1387 cm" (7.21 u) and basepoint 1351 cm" (7.40 ji).
-------
Metobromuron EPA-1
(Tentative)
An alternate peak at 1305 cm" (7.66 yu) with the same
basepoint could be used. Both give a linear absorption curve
over the 3-13 mg/ml range.
Calculation;
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent metobromuron as
follows:
„ m (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method submitted by Eva Santos, EPA Region IX, San Francisco,
California.
David Persch, EPA Region II, New York, N. Y. submitted a similar
method using:
scan range: 2000 cm to 1430 cm (5.0 p to 7.0 yu)
analytical peak: 1683.5 cm (5.94 ji)
_i
basepoint: 1818 cm (5.5 yu)
The absorption curve is linear for 2-16 mg/ml.
Comments on these analytical bands (or others) are most welcome.
-------
November 1975 Metobromuron EPA-2
(Tentative)
Determination of Metobromuron
by Gas-Liquid Chromatography (FID)
Metobromuron is the accepted common name for 3-(p-bromophenyl)-l-
methoxy-1-methylurea, a registered herbicide having the chemical
structure:
0—CH3
Molecular formula: C H BrNLO
Molecular weight: 259
Melting point: 95.5 to 96°C
Physical state and color: white crystalline solid
Solubility: 330 ppm in water at RT; very soluble in acetone,
chloroform, ethanol
Stability: stable; non-corrosive; good compatibility
Other names: Patoran (CIBA), C-3126
Reagents:
1. Metobromuron standard of known % purity
2. Acetone, pesticide or spectro grade
-------
Metobromuron EPA-2
(Tentative)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 2' x 4 mm ID glass column packed with 2% SE-52
on 70/80 mesh Anakrom ABS (or equivalent column)
3. Precision liquid syringe: 5 or 10 jil
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 165°C
Injection temperature: 200°C
Detector temperature: 200°C
Carrier gas: Nitrogen
Carrier gas pressure: 40 psi
Hydrogen pressure: 20 psi
Air pressure: 30 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.1 gram metobromuron standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml acetone by
pipette, close tightly, and shake to dissolve, (final cone
10 mg metobromuron/ml)
-------
Metobromuron EPA-2
(Tentative)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.5 gram metobromuron
into a glass-stoppered flask or screw-cap bottle, add 50 ml acetone
by pipette, close tightly, and shake for one hour. Allow to settle;
filter or centrifuge if necessary, taking precautions to prevent
evaporation, (final cone 10 mg metobromuron/ral)
Determination;
Using a precision liquid syringe, alternately inject three 3-4 ji
portions each of standard and sample solutions. Measure the peak
height or peak area for each peak and calculate the average for both
standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the percent
metobromuron as follows:
» m (pk. ht. or area sample)(wt. std injected)(% purity std)
(pk. ht. or area std)(wt. sample injected)
This method is based on a modification of EPA's Experimental Method
(No. 47) which was adapted from a method from Ciba. Comments, sug-
gestions, data, results, etc. on this method are most welcome.
-------
ovember 1975 Metobromuron EPA-3
(Tentative)
Determination of Metobromuron
by Gas-Liquid Chromatography
(TCD - Internal Standard)
Metobromuron is the accepted common name for 3-(p-bromophenyl)-l-
methoxy-1-methylurea, a registered herbicide having the chemical
structure:
Molecular formula: C_H BrN-0-
Molecular weight: 259
Melting point: 95.5 to 96°C
Physical state and color: white crystalline solid
Solubility: 330 ppm in water at RT; very soluble in acetone,
chloroform, ethanol
Stability: stable; non-corrosive; good compatibility
Other names: Patoran (CIBA), C-3126
-------
2 Metobromuron EPA-3
(Tentative)
Reagents:
1. Metobromuron standard of known % purity
2. Aldrin standard of known HHDN content
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.2 gram HHDN into a
25 ml volumetric flask, dissolve in, and make to volume
with acetone, (cone 8 mg HHDN/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 4' x 1/4" O.D. glass column packed with 4% SE-30
on 60/80 mesh Diatoport S (or equivalent column)
3. Precision liquid syringe: 10 or 25 fil
4. Usual laboratory glassware
Operating Conditions for TCP;
Column temperature: 165°C
Injection temperature: 200°C
Detector temperature: 200°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas pressure: 30-40 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Metobromuron EPA-3
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.05 gram metobrorauron standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 10 ml of
the internal standard solution and shake to dissolve, (final
cone 5 mg metobromuron and 8 mg HHDN/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.05 gram metobromuron
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 10 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the metobromuron.
For coarse or granular materials, shake mechanically for 30
minutes or shake by hand intermittently for one hour, (final
cone 5 mg metobromuron and 8 mg HHDN/ml)
Determination;
Inject 5-15 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is metobromuron, then HHDN.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of metobromuron and HHDN
from both the standard-internal standard solution and the sample-
internal standard solution.
-------
Metobromuron EPA-3
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. HHDN)(% purity HHDN)(pk. ht. or area metobromuron)
(wt. metobromuron)(% purity metobromuron)(pk. ht. or area HHDN)
Determine the percent metobromuron for each injection of the
sample-internal standard solution as follows and calculate the
average:
„ m (wt. HHDNH% purity HHDN)(jpk. ht. or area metobromuron)
" (wt. sample)(pk. ht. or area HHDN)(RF)
This method is based on EPA Experimental Method No. 47B submitted by
G. Radan, EPA, Region II, New York, N. Y. Some changes have been
made in this write-up; therefore, any comments, criticisms, suggestions,
data, etc. concerning this method will be appreciated.
-------
October 1975
MH EPA-1
Determination of MH in Water-Soluble
Formulations by Ultraviolet Spectroscopy
MH is the common name for 1,2-dihydro-pyridazinedione, a
registered growth retardant and selective herbicide having the
chemical structure:
H—C
H—C
Molecular formula: C.H.N.O.
4422
Molecular weight: 112.1
Melting point: 296 to 298°C; the technical product is at least
97% pure and has a m.p. of at least 292°C.
Physical state, color, and odor: odorless, white crystalline powder
Solubility: at 25°C is 0.6% in water, 0.1% in ethanol or acetone,
2.4% in dimethylfonnamide
Stability: stable to hydrolysis; decomposed by strong acids with
release of nitrogen. Behaves as a mono-basic acid and
forms salts with alkali metals and amines; these salts
are water-soluble but are precipitated by hard water.
Other names: Maleic hydrazide; MH-30 (Uniroyal); Retard (Ansul); De-Cut;
De-Sprout; Regulox; Royal MH-30; Slo-Gro; Sprout-Stop;
Stuntman; Suckerstuff; Vonaldehyde; Vondrax; KMH, Maintain 3;
1,2,3,6-tetrahydro-3,6-dioxo-pyridazine; 6-hydroxy-3-(2H)-
pyridazinone
-------
2 MH EPA-1
Reagents;
1. MH standard of known % purity
2. Sodium hydroxide, approx. 0.1N (freshly prepared)
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Usual laboratory glassware
Procedure;
Preparation of Standard:
Weigh 0.1 gram MH into a 250 ml volumetric flask; dissolve
in and make to volume with 0.1N sodium hydroxide solution. Mix
thoroughly and pipette 5 ml into a 100 ml volumetric flask, make
to volume with 0.1N sodium hydroxide solution, and mix thoroughly.
(final cone 20 ug/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.1 gram MH into a
250 ml volumetric flask, make to volume with 0.1N sodium hydroxide
solution,and mix thoroughly. Pipette a 5 ml aliquot into a 100 ml
volumetric flask and make to volume with 0.1N sodium hydroxide
solution. (final cone 20 jig MH/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 330 nm with 0.1N
sodium hydroxide solution in each cell. Scan both the standard
and sample from 360 nm to 280 nm with 0.1N sodium hydroxide
solution in the reference cell. Measure the absorbance of both
standard and sample at 330 nm.
-------
MH EPA-1
Calculation;
From the above absorbances and using the standard and
sample concentrations, calculate the percent MH as follows:
-/ = (abs. sample) (cone, std in ug/ml) (% purity std)
(abs. std) (cone, sample in jig/ml)
-------
August 1975 Monocrotophos EPA-1
Determination of Monocrotophos
by Infrared Spectroscopy
Monocrotophos is the common name for dimethyl phosphate of
3-hydroxy-N-methyl-cis-crotonamide, a registered insecticide having
the chemical structure:
CH3-0
P— 0— C=C — C — N— CH-
II
0
Molecular formula: C_H ,NO P
Molecular weight: 223
Melting point: 54 to 55°C (technical material 25 to 30°C)
Physical state, color, and odor: colorless to white crystalline
material with a mild ester odor. The technical
product is a reddish brown semi-solid.
Solubility: miscible with water; soluble in acetone and ethanol;
sparingly soluble in xylene but almost insoluble in
diesel oils and kerosene
Stability: unstable in lower but stable in higher alcohols and
glycols, stable in ketones; hydrolyzes slowly at
pH 1 to 7, rapidly above pH 7; corrosive to black iron,
drum steel, brass, SS 304, but does not attack glass,
aluminum, or SS 316; incompatible with alkaline pesticides
Other names: Azodrin (Shell); Nuvacron (Ciba); Monocron; dimethyl-1-
methyl-2-methyl-carbamoyl-vinyl phosphate; cis-3-
(dimethoxyphosphinyloxy)-N-methylcrotonamide; 0, 0-
dimethyl-0-(2 methylcarbamoyl-1-^nethyl-vinyl)-phosphate
-------
2 Monocrotophos EPA-1
Reagents;
1. Monocrotophos standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Anhydrous sodium sulfate, granular
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.1 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard;
Weigh 0.2 gram monocrotophos standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml chloroform by
pipette, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 20 mg/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.2 gram monocrotophos
into a glass-stoppered flask or screw-cap tube. Add 10 ml chloro-
form by pipette and 1-2 grams anhydrous sodium sulfate. Close
-------
3 Monocrotophos EPA-1
tightly and shake for one hour. Allow to settle; centrifuge or
filter if necessary, taking precaution to prevent evaporation.
(final cone 20 mg monocrotophos/ml)
Determination;
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 945 cm to
870 cm"1 (10.6 p to 11.5 ju).
Determine the absorbance of standard and sample using the
peak at 900 cm (11.1 ji) and basepoint at 920 cm" (10.86 p).
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent monocrotophos as
follows:
(abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg monocrotophos/ml chloroform gives an
absorbance of approx. 0.009 in a 0.1 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
-------
October 1975
Monocrotophos EPA-2
Determination of Monocrotophos
by Gas-Liquid Chromatography
(FID - Internal Standard)
Monocrotophos is the common name for dimethyl phosphate of
3-hydroxy-N-methyl-cis-crotonamide, a registered insecticide having
the chemical structure:
CH3—
J
P— 0— C
CH3
C— C— N— CH3
II
0
Molecular formula: C-.H,.NO,.P
7 14 5
Molecular weight:
Melting point:
223
54 to 55°C (technical material 25 to 30°C)
Physical state, color, and odor: colorless to white crystalline
material with a mild ester odor. The technical
product is a reddish brown semi-solid.
Solubility: miscible with water; soluble in acetone and ethanol;
sparingly soluble in xylene but almost insoluble in
diesel oils and kerosene
Stability: unstable in lower but stable in higher alcohols and
glycols, stable in ketones; hydrolyzes slowly at pH 1
to 7, rapidly above pH 7; corrosive to black iron, drum
steel, brass, SS 304, but does not attack glass, aluminum,
or SS 316; incompatible with alkaline pesticides
-------
2 Monocrotophos EPA-2
Other names: Azodrin (Shell); Nuvacron (Ciba); Monocron; dimethyl-1-
methyl-2-methyl-carbamoyl-vinyl phosphate; cis-3-
(dimethoxyphosphinyloxy)-N-methylcrotonaraide; 0,0-
dimethyl-0-(2 methylcarbamoyl-1-methyl-vinyl)-phosphate
Reagents:
1. Monocrotophos standard of known % purity
2. Methyl parathion standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.75 gram methyl parathion
into a 50 ml volumetric flask, dissolve in, and make to volume
with acetone, (cone 15 mg methyl parathion/ml)
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 5' x 1/8" stainless steel column packed with 3% SE-30
on 100/120 Varaport 30 (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 175°C
Injection temperature: 225°C
Detector temperature: 240°C
Carrier gas: Nitrogen
Carrier gas flow rate: 50 ral/min
Hydrogen flow rate: 30 ml/min
Air flow rate: 300 ml/min
-------
3 Monocrotophos EPA-2
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.1 gram monocrotophos standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 10 ml of
the internal standard solution and shake to dissolve, (final
cone 10 mg monocrotophos and 15 mg methyl parathion/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.1 gram monocrotophos
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the monocrotophos.
For coarse or granular materials, shake mechanically for 10-15
minutes or shake by hand intermittently for 25-30 minutes.
(final cone 10 mg monocrotophos and 15 mg methyl parathion/ml)
Detenninat ion:
Inject 2-3 pi of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is monocrotophos, then methyl
parathion.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of monocrotophos and methyl
parathion from both the standard-internal standard solution and
the sample-internal standard solution.
-------
4 Monocrotophos EPA-2
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
IS = internal standard = methyl parathion
(y-t-_ . IS) (% purity IS)(pk. ht. or area monocrotophos^ _
(wt. monocrotophos) (% purity monocrotophos) (pk. ht. or area IS)
Determine the percent monocrotophos for each injection of the
sample-internal standard solution as follows and calculate the
average :
7 _ (vt- IS)(% purity IS)(pk. ht. or area monocrotophos)
(wt. sample) (pk. ht. or area IS) (RF)
Method submitted by Division of Regulatory Services, Kentucky
Agricultural Experiment Station, University of Kentucky, Lexington,
Kentucky 40506.
-------
September 1975
Monuron EPA-1
Determination of Monuron by
Alkaline Hydrolysis and Titration
Monuron is the common name for 3-(p-chlorophenyl)-l,l-dimethyl-
urea, a registered herbicide having the chemical structure:
Molecular formula: C_H ..C1N-0
Molecular weight: 198.6
Melting point: 174-175'C
Physical state, color, and odor: odorless, white crystalline solid
Solubility: 230 ppm in water at 25°C; sparingly soluble in petroleum
oils and in polar organic solvents; 5.2% in acetone at
27°C
Stability: stable toward moisture and oxidation at RT but is
decomposed at 185-200°C; rate of hydrolysis at RT or
neutrality is negligible but is increased at elevated
temp, or more acid or alkaline conditions; non-corrosive
and non-flammable
Other names: Telvar (DuPont), chlorfenidim (USSR), Monurex
Principle of the Method;
The monuron is hydrolyzed to p-chloroaniline, carbon dioxide
(as carbonate), and dimethylamine. The dimethylamine is distilled and
titrated. Volatile, moderately strong bases, or substances that
hydrolyze to give them, interfere.
-------
2 Monuron EPA-1
Reagents;
1. Potassium hydroxide, 20% solution
2. Hydrochloric acid, 0.1N standard solution
3. Sodium hydroxide, O.lN standard solution
4. Ethyl alcohol, ACS
5. Glycerol, ACS
Equipment;
1. Distilling apparatus consisting of a 500 ml round-bottom
flask with a thermometer well in the side and a 24/40
standard taper (ST) joint at the top. The flask is con-
nected to the bottom of a vertical condenser which has its
top connected to the top of a second vertical condenser by
a horizontal tube with a right angle 24/40 ST joint on
each end. The bottom of the second condenser is connected
by 24/40 ST joint to the top of a delivery tube which has
a narrow plain end extending almost to the bottom of a
receiving beaker.
2. 500 ml size heating mantle with variable transformer control
3. Thermometer to 200°C
4. Potentiometric titrimeter
5. Usual laboratory glassware
Procedure:
Weigh a portion of sample equivalent to 0.4-0.5 gram inonuron into
the reaction flask, dissolve in 25 ml ethyl alcohol, and add 100 ml
glycerol and 100 ml 20% potassium hydroxide solution. Attach
immediately to the first condenser.
-------
Monuron EPA-1
Pipette SO ml of the 0.1N standard hydrochloric acid into the
receiving beaker. Reflux at a moderate rate for 2-1/2 hours with
water flowing through both condensers. Remove the water from the
first condenser and distill until the temperature at the thermometer
well reaches 175°C — usually about 50 minutes. (The temperature
rises rapidly at the end.)
Titration;
Remove the delivery tube and receiving beaker and rinse the
delivery tube into the beaker. Titrate the excess standard acid
with the 0.1N standard sodium hydroxide potentiometrically, using
a glass electrode and a calomel electrode. The inflection point,
which occurs at about pH 7.6, is taken as the endpoint.
With less accuracy, bromthymol blue may be used as an internal
indicator.
Calculation;
Calculate the percentage of monuron as follows:
z m (ml)(N)(0.1986)(100)
(g sample)
where: 0.1986 is the milliequivalent weight of monuron
(1 ml 0.1N HC1 - 0.01986 g monuron)
This method is based on Lowen and Baker, Anal. Chem. 2A, 1475 (1952).
-------
September 1975
Monuron EPA-2
Determination of Monuron
by Ultraviolet Spectroscopy
Monuron is the common name for 3-(p-chlorophenyl)-l,l-dimethyl-
urea, a registered herbicide having the chemical structure:
Molecular formula: C_H -C1N 0
Molecular weight: 198.6
Melting point: 174-175°C
Physical state, color, and odor: odorless, white crystalline solid
Solubility: 230 ppm in water at 25°C; sparingly soluble in petroleum
oils and in polar organic solvents; 5.2% in acetone at
27°C
Stability: stable toward moisture and oxidation at RT but is
decomposed at 185-200°C; rate of hydrolysis at RT or
neutrality is negligible but is increased at elevated
temp, or more acid or alkaline conditions; non-corrosive
and non-flammable
Other names: Telvar (DuPont), chlorfenidim (USSR), Monurex
Reagents:
1. Monuron standard of known % purity
2. Methanol, pesticide or spectro grade
-------
2 Monuron EPA-2
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm silica cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard;
Weigh 0.1 gram monuron standard into a 100 ml volumetric
flask, add 100 ml methanol by pipette, and mix thoroughly.
Pipette 10 ml into a second 100 ml volumetric flask, make to
volume with methanol, and mix thoroughly. Pipette 5 ml into
a third 100 ml volumetric flask, make to volume with methanol,
and mix thoroughly. (final cone 5 ug/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.1 gram monuron
into a 250 ml glass-stoppered or screw-cap flask, add 100 ml
methanol by pipette, and shake on a mechanical shaker for 30
minutes. Allow to settle; centrifuge or filter if necessary,
taking precautions to prevent evaporation. Pipette 10 ml into
a 100 ml volumetric flask, make to volume with methanol, and
mix thoroughly. Pipette 5 ml of this solution into another
100 ml volumetric flask, make to volume with methanol, and mix
thoroughly, (final cone 5 fig monuron/ml)
UV Determination:
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen at 0 and 100% transmission at 245 nm with
-------
3 Monuron EPA-2
methanol in each cell. Scan both the standard and sample from
300 run to 200 nm with methanol in the reference cell.
Measure the absorbance of standard and sample at 245 nm.
Calculat ion:
From the above absorbances and using the standard and sample
concentrations, calculate the percent monuron as follows:
«. (abs. sample) (cone, std in ug/ml)(% purity std)
(abs. std)(cone, sample in jug/ml)
-------
September 1975 Monuron EPA-3
Determination of Monuron
by Infrared Spectroscopy
Monuron is the common name for 3-(p-chlorophenyl)-l,l-dimethyl-
urea, a registered herbicide having the chemical structure:
Molecular formula: C.H C1N.O
Molecular weight: 198.6
Melting point: 174-175°C
Physical state, color, and odor: odorless,white crystalline solid
Solubility: 230 ppm in water at 25°C; sparingly soluble in petroleum
oils and in polar organic solvents; 5.2% in acetone at
27°C
Stability: stable toward moisture and oxidation at RT but is
decomposed at 185-200°C; rate of hydrolysis at RT or
neutrality is negligible but is increased at elevated
temp, or more acid or alkaline conditions; non-corrosive
and non-flammable
Other names: Telvar (DuPont), chlorfenidim (USSR), Monurex
Reagents;
1. Monuron standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
2 Monuron EPA-3
Equipment:
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.5 mm KBr or NaCl cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure;
Preparation of Standard;
Weigh 0.1 gram monuron standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml chloroform by pipette,
close tightly, and shake to dissolve. Add a small amount of
anhydrous sodium sulfate to insure dryness. (final cone 10 mg/ml)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.5 gram monuron
into a glass-stoppered flask or screw-cap bottle. Add 50 ml
chloroform by pipette and 1-2 grams anhydrous sodium sulfate.
Close tightly and shake for one hour. Allow to settle; centri-
fuge or filter if necessary, taking precautions to prevent
evaporation, (final cone 10 mg monuron/ml)
Determination;
With chloroform in the reference cell, and using the optimum
quantitative analytical settings, scan both the standard and
sample from 1400 cm to 1300 cm (7.1 p to 7.7 ja) .
Determine the absorbance of standard and sample using the
peak at 1360 cm" (7.35 ji) and baseline from 1380 cm to 1325 cm"
(7.25 u to 7.55
-------
Monuron EPA-3
Calculation;
From the above absorbances and using the standard and
sample solution concentrations, calculate the percent monuron
as follows:
v m (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
-------
September 1975
Neburon EPA-1
(Tentative)
Determination of Neburon
by Infrared Spectroscopy
Neburon is the accepted common name for l-n-butyl-3-(3,4-
dichlorophenyl)-l-methylurea, a registered herbicide having the
chemical structure:
Cl
0 CH3
N—CH2—CH2—CH2 CH3
Molecular formula: C10H.,C10N00
J./ lo / /
Molecular weight: 275.18
Melting point: 102 to 103°C
Physical state, color, and odor: odorless, white crystalline solid
Solubility: 4.8 ppm in water at 24°C; very low in common hydro-
carbon solvents
Stability: stable toward oxidation and moisture under normal
storage conditions
Other names: Kloben (DuPont), Neburex, neburea (So. Africa)
Reagents;
1. Neburon standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
Neburon EPA-1
(Tentative)
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.5 mm KBr or NaCl cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.08 gram neburon standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml carbon disulfide by pipette,
close tightly, and shake to dissolve. Add a small amount of
anhydrous sodium sulfate to insure dryness. (final cone 8 mg/ml)
Preparation of Sample:
Weigh an amount of sample equivalent to 0.4 gram neburon
into a glass-stoppered flask or screw-cap bottle. Add 50 ml
carbon disulfide by pipette and 1-2 grams anhydrous sodium sulfate.
Close tightly and shake for one hour. Allow to settle; centrifuge
or filter, taking precaution to prevent evaporation, (final cone
8 mg neburon/ml)
Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan both the standard and sample from
1430 cm'1 to 1175 cm'1 (7.0 ^t to 8.5 /u).
Determine the absorbance of the standard and sample using the
peak at 1289 cm (7.76 ji) and basepoint 1319 cm" (7.58/u).
-------
Neburon EPA-1
(Tentative)
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent neburon as
follows:
2 - (abs. sample)(cone. std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method contributed by Eva Santos, EPA Region IX, San Francisco,
California.
-------
February 1976
Nitrophenols EPA-1
Determination of Nitrophenols in Formulations
by Stannous Chloride Reduction
Nitrophenols are those compounds having one or more nitro groups
on a phenol. These compounds may be registered as acaricides, fungi-
cides, herbicides, or insecticides. The chemical structure is similar
to that of 4,6-dinitro-o-cresol which is:
OH
CH3
4,6-dinitro-o-cresol has the common name DNOC and the following
characteristics:
Molecular formula: C_H,N-0_
/ D 2. D
Molecular weight: 198.1
Melting point: 86°C
Physical state, color, and odor: yellowish, odorless, crystals
Solubility: 130 ppm in water at 15°C; soluble in most organic solvents
and in acetic acid; alkali salts are water-soluble; technical
grade is 95-98% pure and has a mp 83 to 85°C
Stability: explosive, therefore it is usually moistened with up to 10%
water to reduce the hazard; corrosive to mild steel in the
presence of moisture
-------
2 Nitrophenols EPA-1
Principle of the Method:
A volume of stannous chloride solution in excess of that needed by
a weighed portion of sample is titrated with standard potassium dichromate
solution without reacting it with the sample. A second identical portion
is reacted with the sample and the excess titrated. The difference in
titrations represents the amount of potassium dichromate equivalent to
the sample. Other oxidizing compounds, reducible by stannous chloride,
are titrated with standard sodium thiosulfate and are subtracted as
milliequivalents from the dichromate milliequivalents of sample. The
net milliequivalents are equal to the nitro phenolic compound in the
sample.
Reagents:
1. Potassium dichromate, 0.3N standard solution - weigh 14.71 grams
pure potassium dichromate (previously dried 2 hr at 100°C)
into a one liter volumetric flask, dissolve in,and make to volume
with distilled water.
2. Stannous chloride solution - weigh 17 grams stannous chloride
dihydrate into a 500 ml volumetric flask, dissolve in,and make
to volume with 18-19% hydrochloric acid (1+1 by weight). The
strength of this solution is approximately equivalent to the
dichromate solution but weakens gradually upon oxidation.
3. Glacial acetic acid, reagent grade
A. Concentrated hydrochloric acid
5. Potassium iodide, 15% solution in water
6. Starch indicator solution
7. Sodium thiosulfate, 0.1N (or 0.3N) standard solution
-------
Nitrophenols EPA-1
Equipment :
1, 300 ml Erlenmeyer flask with rubber stopper fitted with a
Bunsen valve (described below)
-A
-D
The Bunsen valve is a short 2-4"
length of rubber tubing (A) stoppered
at one end (B) and fitted over a piece
of glass tubing (C) at the other end.
A 1/2-3/4" slit (D) is made with a
razor blade along the length of the
tubing. This slit allows internal
pressure to be relieved by allowing
gases to escape, but is sealed as out-
side pressure pushes in since the sides
of the slit are pressed together.
2. Water bath, 95— 100°C
3. Usual laboratory glassware and titration apparatus
Procedure : (written for dinitrocresol)
Weigh a portion of sample equivalent to 1.3-1.7 grams dinitrocresol
into a 250 ml volumetric flask, dissolve in, and make to volume with dis-
tilled water.
Pipet a 10 ml aliquot of sample solution into a 250 ml Erlenmeyer
flask, add 5 ml glacial acetic acid, 8 ml concentrated hydrochloric acid,
and, by pipet, 25.0 ml stannous chloride solution. Close flask with stopper
fitted with a Bunsen valve and heat on a water bath at 95 — 100°C for 30
minutes. Cool by immersing in cold water and dilute to about 200 ml with
distilled water. Add 3 ml of 15% potassium iodide solution and 1 ml
starch indicator solution. Titrate with 0.3N potassium dichromate solution
with constant agitation to a blue end point. (If the end point is passed,
-------
4 Nitrophenols EPA-1
the slight excess of dichromate may be back-titrated with sodium thio-
sulfate.)
Determine the dichromate equivalent of 25.0 ml stannous chloride by
repeating the above procedure,omitting the sample. Heating is not nec-
essary, but would more closely match the sample determination conditions.
The difference in the two dichromate titrations is equal to the dinitro-
cresol in the sample aliquot and any other oxidizing compounds, reducible
by stannous chloride.
To determine the amount of other oxidizing compounds: take a 10 ml
aliquot of sample solution, add 3 ml 15% potassium iodide solution, 5 ml
glacial acetic acid, 1 ml starch indicator, 200 ml water, and titrate
with 0.1N sodium thiosulfate solution to the disappearance of the blue
color.
Calculations:
The ml dichromate used for 25 ml SnCl (blank) minus the ml dichromate
used for 25 ml SnCl. plus 10 ml sample solution (sample) multiplied by the
normality of the dichromate (N) equals the milliequivalents (meqs.) of
dinitrocresol (DNOC) and other oxidizing compounds (Ox cmpds.).
(Blank - sample)(N) = meqs. DNOC + Ox cmpds.
The ml thiosulfate multiplied by the normality equals the milliequiv-
alents of other oxidizing compounds which is subtracted from the above to
give the milliequivalentsof DNOC in 10 ml of sample aliquot.
„ ,, . . . n (meqs. of DNOC in 10 ml)(100)
/, dinitrocresol in sample = -,—3 :—\ /-,'n /"o'cn\
v (grams sample)(10/250)
-------
Nitrophenols EPA-1
Chemical Reactions:
Bichromate equivalent of stannous chloride:
6SnCl
f\ ' *• *\ *^ *• O T
+ 6KI + 14HC1
28HC1
6SnCl4 + 4KC1 + ACrCl + 14H 0
8KC1 + 2CrCl + 31 +
Sample reaction with stannous chloride:
HO
•N02
-h6S-nCI2 -I-
N02
NH-HCI
6S>»CI4 -h 4H20
NH-HCI
Oxidizing compounds with sodium thiosulfate
(oxidizing compounds) + KI
2NaI + Na-S.O,
-------
February 1976
Nitrophenols EPA-2
Determination of Nitrophenols in Formulations
by Total Nitrogen
Nitrophenols are those compounds having one or more nitro groups
on a phenol. These compounds may be registered as acaricides, fungi-
cides, herbicides, or insecticides. The chemical structure is similar
to that of 4,6-dinitro-o-cresol which is:
OH
4,6-dinitro-o-cresol has the common name DNOC and the following
characteristics:
Molecular formula: C,H,N_0C
/ D 2. J
Molecular weight: 198.1
Melting point: 86°C
Physical state, color, and odor: yellowish, odorless, crystals
Solubility: 130 ppm in water at 15°C; soluble in most organic solvents
and in acetic acid; alkali salts are water-soluble; tech-
nical grade is 95-98% pure and has a mp 83 to 85°C
Stability: explosive, therefore it is usually moistened with up to
10% water to reduce the hazard; corrosive to mild steel
in the presence of moisture
-------
2 Nitrophenols EPA-2
Principle of the Method:
These compounds may be in dusts, wettable powders, emulsifiable
concentrates, oil sprays, or as 98-100% free acid flakes. If there
are no interfering nitrogen-containing constituents present, they may
be determined directly from total nitrogen; otherwise, an extraction
clean-up procedure is necessary.
Since the nitrogen is present in the nitro (oxidized) form, it
must be converted to the amino (reduced) form before being determined
by the regular Kjeldahl procedure. This is done by reacting the sample
with acetic acid-zinc dust and salicylic acid-sodium thiosulfate. These
are the agents which reduce the nitro (-NCL) to amino (-NH ) so that it
may be reduced to ammonium sulfate by the sulfuric acid regular Kjeldahl
procedure.
Reagents;
1. Acetone
2. Concentrated hydrochloric acid
3. 50% ethyl alcohol-water (1+1)
4. Potassium hydroxide solution (1+1)
5. Ethyl ether
6. Petroleum ether
7. Acetic acid, glacial
8. Zinc dust
9. Sulfuric acid (1+4)
10. Sodium thiosulfate
11. Concentrated sulfuric acid, reagent grade
-------
3 Nitrophenols EPA-2
12. Salicylic acid, reagent grade
13. Zinc dust, reagent grade
14. Mercuric oxide, red, reagent grade
(Commercial packages called "Kel-pacs" are available containing
various oxidizing catalysts and various amounts of potassium
sulfate in small oxidizable plastic packets. One packet can be
dropped into the flask,saving the weighing and transfer of the
HgO and K^SO^)
15. Potassium sulfate, reagent grade (see above)
16. Sodium or potassium sulfide, reagent grade
17. Granulated zinc, reagent grade
18. Kjeldahl sodium hydroxide solution (450 grams NaOH,free from
nitrates,in one liter of water)
19. Phenolphthalein indicator solution
20. Sulfuric acid, 0.1N standard solution
(An alternative procedure is to use 50 ml of a saturated boric
acid solution that simply holds the ammonia which is titrated
with standard acid. The procedure eliminates the need for
standard alkali solution.)
21. Sodium hydroxide, 0.1N standard solution (see above)
22. Mixed methyl red indicator solution - dissolve 1.25 grams
methyl red and 0.825 gram methylene blue in one liter of 90%
ethyl alcohol. The color change is from purple in acid to
green in basic solution.
Equipment:
1. Filtration equipment
2. Steam bath
3. 800 ml Kjeldahl flask
-------
Nitrophenols EPA-2
4. Kjeldahl digestion and distillation apparatus
5. Titration apparatus
6. Usual laboratory glassware
Procedure:
Extraction-cleanup procedure:
If it is known that no interfering nitrogen-containing con-
stituents are present, omit the following extraction cleanup
procedure and begin directly with the nitrogen determination.
Weigh an amount of sample equivalent to 0.025-0.30 gram of
nitrogen into a 200 ml volumetric flask. Add approximately 100 ml
acetone and sufficient concentrated hydrochloric acid to make
distinctly acid. Make to volume and shake intermittently over
several hours. (If the amount of dust or powder is large, correct
for its volume by adding the same weight to 200 ml acetone in an
identical volumetric flask and note the increase above the line—
adjust the sample flask to the same amount.)
Filter if necessary and pipette 100.0 ml of the clear liquid
into a beaker or flask. Evaporate on a steam bath to remove the
acetone. Add 50 ml of 50% ethyl alcohol and make alkaline to
phenolphthalein with aqueous potassium hydroxide solution (1+1).
Digest on a steam bath 10-15 minutes and cool. If oils are present,
extract with petroleum ether. Filter and wash filter paper thor-
oughly with 50% alcohol. Evaporate most of the filtrate on a steam
bath to remove the alcohol. Cool, transfer to a separatory funnel
with water, and acidify with hydrochloric acid. Extract with ethyl
ether three times, using each time a volume of ether eoual to the
volume of aqueous solution in the separatory funnel. Combine the
ether extracts into a second separatory funnel and wash once with
water acidified with HC1.
-------
5 Nitrophenols EPA-2
Reduction of NO Group:
Transfer the. ether into an 800 ml Kjeldahl flask and evaporate
on a steam bath to just dryness. Dissolve the residue in 5 ml
acetic acid, add 1 gram zinc dust, mix, and heat on a steam hath
for 15 minutes. Add 1 ml sulfuric acid (1+4) and let stand over-
night at room temperature. In the morning, add another 1 ml sul-
furic acid (1+4) and heat on a steam bath for 15 minutes. Cool,
add 35 ml concentrated sulfuric acid containing 2 grams salicylic
acid, allow to stand a few minutes, add 5 grams sodium thiosulfate,
and heat over a low flame until most of the sulfur dioxide is
expelled.
Digestion:
Add 0.7 gram mercuric oxide and 10 grams potassium sulfate (or
one Kel-pac) and continue boiling until the liquid in the flask has
been colorless for one hour. If the contents of the flask tend to
become solid before this point is reached, add 10 ml more of sul-
furic acid. To avoid decomposition of ammonium sulfate and subse-
quent loss of ammonia,do not allow the flame to reach any part of
the flask not in contact with liquid. The flask may be lifted from
the digestion rack and the acid swirled around the inside of the
flask to wash undigested particles back into the acid. When digestion
is complete, cool; add 200 ml-300 ml water, making sure that the
digestion mixture is completely dissolved.
Distillation;
Measure 50.00 ml of standard 0.1N sulfuric acid into a 500 ml
Erlenmeyer wide-mouth flask, add several drops of mixed methyl red
indicator solution, and place under the condenser of the distilling
apparatus, making sure that the condenser tube extends beneath the
surface of the acid in the flask. A glass tube attached by inert
tubing to the condenser outlet tube is very convenient when later
-------
6 Nitrophenols EPA-2
removing the receiving flask. If the indicator changes from acidic
(purple) to basic (green), the determination must be repeated using
less sample or more acid in the receiving flask.
Add 25 ml sodium or potassium sulfide solution and mix thor-
oughly; then add several pieces of granulated zinc.
(When using mercury as a catalyst, it must be
precipitated with K or Na sulfide before the
distillation process since it forms a complex
substance with ammonia which is not readily
decomposed by alkali.)
(Zinc in an alkaline solution slowly reacts to
form a zincate and hydrogen: Zn + 2NaOH — ) Na ZnO? + H_
This slow evolution of hydrogen keeps the solution
stirred, thereby preventing superheating.
Pour about 110 ml of the Kjeldahl sodium hydroxide solution (or
if extra acid was added, use 25 ml more alkali for each 10 ml acid
added) slowly down the inclined neck of the flask so that it layers
under the acid solution without mixing. A few drops of phenol-
phthalein may be added to be sure sufficient alkali is added to
neutralize all the acid, remembering that a considerable excess of
alkali will destroy the pink color.
Connect the flask to the condenser by means of a Kjeldahl con-
necting bulb, ignite the burner, and quickly mix the contents of
the flask thoroughly with a rotary motion. It is advisable to begin
the distillation with a small flame until the solution begins to
boil, then increase the heat until the solution boils briskly. Dis-
till 150-200 ml of the liquid (the first 150 ml usually contains all
of the ammonia) into the receiving flask. Move the flask so that the
tip of the delivery tube is above the level of the liquid and distill
another 10 ml or so to wash the inside of the tube. Shut off heat,
wash the outside of the delivery tube, and remove flask from apparatus.
-------
Nitrophenols EPA-2
Titration and Calculation:
Titrate the excess standard acid with standard 0.1N sodium
hydroxide using mixed methyl red indicator. Reagents for this
determination should be acid-free or a reagent blank should be
run. Calculate the percent nitrogen as follows:
Using a blank:
= (ml NaOH for blank—ml NaOH for sample) (N of NaOH)(.01401)(100)
(grams of sample)*
Not using a blank:
[(ml H SO.)(N of H SO )~(ml NaOH) (N of NaOH) ]( .01401) (100)
_ *y 4 _ 4-, - _
(grams of sample)*
» ,,. , , . , % nitrogen in sample
% Nitrophenolic compound = ~-—rr— : 7—' :
1 l / »^ T r- Tf r\.r*n.Tf\ ~\ v\ Y^^^-W**"»TN
% nitrogen in nitrophenolic compound
* If extraction-cleanup procedure was used, a dilution factor of
100/200 must be added here.
-------
November 1975
Norbormide EPA-1
Determination of Norbormide in Baits
by Ultraviolet Spectroscopy
Norbormide is the accepted common name for 5-(alpha-hydroxy-alpha-
2-pyridylbenzyl)-7-(alpha-2-pyridylbenzylidene)-5-norbornene-2,3-
dlcarboximide, a registered rodenticide having the chemical structure:
N
N—H
Molecular formula: C33H25N3°3
Molecular weight: 511.6
Melting point: 180 to 190°C (190 to 198° on crystals from methylene
chloride 4- ether)
Physical state and color: white to off-white crystalline powder
(mixture of isomers)
Solubility: 60 ppm in water at RT; at 30°C solubility in 100 ml is
1.4 mg in ethanol, 15 mg in chloroform, 0.1 mg in ether,
2.9 mg in 0.1N HC1; soluble in dilute acids
Stability: stable at RT when dry, and to boiling water; hydrolyzed by
alkali; non-corrosive
Other names: Shoxin, Raticate (McNeil Laboratories)
-------
2 Norbormide EPA-1
Reagents:
1. Norbormide standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
4. Decolorizing carbon (Norit A or equivalent)
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm silica cells
2. Mechanical shaker
3. Filtration apparatus
A. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.1 gram norbormide standard into a 100 ml volumetric
flask, dissolve, make to volume with chloroform, and mix thor-
oughly. Pipette 2 ml into a second 100 ml volumetric flask and
make to volume with chloroform. (final cone 20 ug/ml)
Preparation of Sample:
Weigh an amount of sample equivalent to 0.02 gram norbormide
into a 250-300 ml glass-stoppered flask, add 2 grams anhydrous
sodium sulfate and 2 grams decolorizing carbon (Norit A or equiv-
alent) , pipette in 100 ml chloroform, and shake on a mechanical
shaker for one hour. Filter a portion of the chloroform extract
through a coarse, soft, rapid filter paper, taking precautions
against solvent loss by evaporation. Pipette 10 ml of clear
filtrate (discard the first few ml coming through the paper) into
a 100 ml volumetric flask and make to volume with chloroform.
(final cone 20 ig norbormide/ml)
-------
3 Norbormide EPA-1
UV Determination:
Using the optimum quantitative settings for the particular
UV instrument being used, adjust the 0 and 100% settings at
253 nm with chloroform in both cells. Scan both standard and
sample from 300 nm to 200 nm.
Calculation:
Measure the absorbance of standard and sample at 253 nm and
calculate the percent norbormide as follows:
7 - (abs. sample)(cone, std in jug/ml)(% purity std)
(abs. std) (cone, sample in jig/ml)
or using dilution factors, as follows:
_ (abs. sample)(wt. std)(purity std)(1/100)(2/100)(100)
" (abs. std)(wt. sample)(1/100)(10/100)
-------
November 1975
Oil of Lemongrass EPA-1
(Tentative)
Determination of Oil of Lemongrass by
Gas-Liquid Chromatography (TCD)
Oil of Lemongrass is a registered animal repellent consisting of
75-85% citral as the active constituent. Citral is 3,7-dimethyl-2,6-
octadienal which occurs in two geometric isomers with chemical
structures as follows:
geranial (citral a)
•CHO
neral (citral b)
OHO
Molecular formula: C,rtH,,0
1U ID
Molecular weight: 152.23
geranial is a light oily liquid with a strong lemon odor; b.p._ , 92-93*C;
** / .O
2Q 20
d7 0.8888; n' 1.48982; practically insoluble in water; miscible
with alcohol, ether, benzyl benzoate, diethyl phthalate, glycerol,
propylene glycol, mineral oil, essential oils
neral is a light oily liquid; lemon odor not as intense but sweeter
?n ?n
than geranial; b.p.2 6 91-92°; dj 0.8869; n^ 1.48690; solu-
bilities same as geranial
-------
Oil of Lemongrass EPA-1
(Tentative)
Stability: unstable to alkalis and strong acids; will cause discolora-
tion of white soaps and alkaline cosmetics
Other names: Lemongrass oil, oil of verbena (Indian)
Note - oil of lemongrass is also used in the synthesis of vitamin A;
as a flavor in fortifying lemon oil; in perfumery for citrus
effect in lemon and verbena scents, in cologne odors, in
perfumes for colored soaps.
This method is based on the thermal conductivity detection of both
isomers of citral using a 20% SE-30 column. See note at end of method
for alternative procedures.
Reagents:
1. Oil of Lemongrass standard of known citral content
2. Acetone, pesticide or spectro grade
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 5' x 1/4" O.D. aluminum column packed with 20% SE-30
on 60/80 mesh Chromosorb W AW DMCS (or equivalent
column)(SS or glass is preferred to Al)
3. Precision liquid syringe: 25 or 50 fil
A. Usual laboratory glassware
-------
3 Oil of Lemongrass EPA-1
(Tentative)
Operating Conditions for TCP;
Column temperature: 150°C
Injection temperature: 250°C
Detector temperature: 250°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas flow rate: 100 ml/min
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
Procedure;
Preparation of Standard;
Weigh 0.6 gram oil of lemongrass standard into a 10 ml volu-
metric flask and make to volume with acetone, (cone 60 ug/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.6 gram oil of lemon-
grass into a 10 ml volumetric flask and make to volume with acetone.
(cone 60 pg oil lemongrass/ml)
For the analysis of aerosols some care must be used in removing
the freons. The chilled sample should be allowed to warm to room
temperature and then heated gently to about 40-50°C just until the
freons are removed. This will minimize loss of any volatile con-
stituents from the oil of lemongrass.
-------
Oil of Lemongrass EPA-1
(Tentative)
Determination:
Using a precision liquid syringe, alternately inject three
20-30 jil portions each of standard and sample solutions, allowing
sufficient time between injections for all sample constituents
to clear the column.
Calculation:
Measure and combine the area of both citral peaks (citral a
and citral b) for both the standard and sample. Using the average
of several injections, calculate the % oil lemongrass as follows:
„ m (pk. area sample)(wt. standard injected)(% purity standard)
(pk. area standard)(wt. sample injected)
(If sample was an aerosol, multiply above result by the %
nonvolatile.)
The above method is basically that developed by Margaret Frost and
Mario V. Conti, EPA, Region IX, San Francisco, Calif. A few changes
were made and some additional information obtained at EPA's Beltsville
Chemistry Laboratories was added in this write-up; therefore, any sug-
gestions, data, or criticisms are most welcome.
Note on alternative procedures;
Frost and Conti have also successfully used a 10% Carbowax 20 M
column and a 10% QF-1 column both at 155* using a thermal conductivity
detector.
Ronald F. Thomas, EPA, Beltsville, Md., has used a 1/8" x 5' pyrex
10% Carbowax 20 M 60/80 Chromosorb W AW column at 105°C with a flame
ionization detector and nitrogen for carrier gas.
-------
January 1976 Organotin Compounds EPA-1
Determination of Tin in Organotin Compounds
by Oxidation, Reduction, and Titration
Several tin-based organic compounds are registered fungicides,
bactericides, algicides, and molluscicides. These compounds are of
two main types:
(a) tributyltin or triphenyltin compounds:
example: tributyltin acetate
0 CH2-CH2-CH2-CH3
CH3—C—0—Sr>A-CH2-CH2-CH2—CH3
\H2-CH2-CH2—CH3
(b) bis (tributyltin) compounds:
example: bis (tributyltin) oxide
CH3-CH2 —CH2-CH2 CH2-CH2-CH2-CH3
CH3-CH2—CH2-CH2ASn—0—Sti/CH2 —Ote—CH2-CH-?
/ \
CH3-CH2-CH2-CH2/ CH2—CH2—CH2-CH3
In general, these compounds are practically insoluble in water
but are miscible with organic solvents. Some are solids and some are
liquids. The stability of these compounds varies but most are stable
when dry and stored in dark, closed containers. Most are compatible
with common pesticides, but not with liquids or oil emulsions.
-------
2 Organotin Compounds EPA-1
Principle of the Method;
The organotin compound is digested with sulfuric and nitric acids,
reduced with nickel and iron, and titrated with potassium iodate and
starch as elemental tin. This is then calculated to the specific
organotin compound.
Reagents;
1. Tin standard, pure foil
2. Sulfuric acid, concentrated, ACS
3. Nitric acid, concentrated, ACS
4. Distilled water, boiled and cooled to remove oxygen
5. Hydrochloric acid, (1+2) in water
6. Nickel coil - roll a 6" x 3" x 0.15 (or 0.25)" sheet of pure
nickel into a 3" long roll. Clean before each
use by boiling in (1+2) hydrochloric acid.
7. Iron powder
8. Sodium bicarbonate, saturated solution
9. Dry ice
10. Starch indicator solution, 1% prepared fresh
11. Potassium iodide, 10% solution
12. Potassium iodate, 0.1N standard solution - Prepare and
standardize as described under procedure.
Equipment;
1. Kjeldahl flask and digestion set-up
2. Hot plate
-------
Organotin Compounds EPA-1
3. 500 ml Erlenmeyer with a rubber stopper into which a 7 mm piece
of glass tubing is fitted; the glass tubing is bent to extend
from just below the stopper on the inside, up, over, and down
on the outside to a level just above the bottom of the flask.
(The drawing below shows the shape of the tubing and how it is
extended into a beaker of saturated sodium carbonate during the
cooling operation.)
RUBBER
STOPPER
NICKEL COIL
GLASS TUBE
SAT. NaHCO,
WATER BATH
4. Water bath (or ice bath)
5. Titration apparatus
6. Usual laboratory glassware
Procedure:
Preparation of 0.1N Potassium lodate Solution;
Weigh 3.567 grams potassium iodate and 10 grams potassium iodide,
place in a one-liter volumetric flask, add one pellet of potassium
hydroxide, dissolve in, and make to volume with oxygen-free water.
Place 0.25 gram pure tin foil (accurately weighed) into a 500 ml
Erlenmeyer flask and dissolve in 100 ml concentrated hydrochloric
acid. Add 180 ml oxygen-free water and 10 ml concentrated sulfuric
acid.
Proceed following the same reduction and titration procedure as
for sample. Calculate the normality as shown under calculations.
-------
A Organotin Compounds EPA-1
Preparation of Sample:
Weigh a portion of sample equivalent to about 0.2 gram tin into
a 500 ml (or 800 ml) Kjeldahl flask, add 10 ml concentrated sulfuric
acid, and,cautiously, 20 ml concentrated nitric acid. Place flask on
an asbestos mat with a 2" hole and heat with a small flame at first
until any vigorous reaction subsides. Increase the heat and digest
until white fumes of sulfuric acid are evolved. If the solution
darkens or chars, add more concentrated nitric acid until the
solution remains colorless or a pale yellow. Cool, add 25 ml
water,and heat again to white fumes to expel any oxides of nitrogen.
Cool, add 80 ml water, and transfer to a 500 ml Erlenmeyer flask.
Rinse the Kjeldahl flask with 100 ml water and add to the 500 ml
Erlenmeyer flask. Add 100 ml concentrated hydrochloric acid and
proceed under reduction.
Reduction;
Treat both the standard tin solution and the digested sample
solution as follows:
Add a nickel coil (previously washed) and 5 grams iron powder.
Place the rubber stopper fitted with the glass tubing (as described
under equipment) tightly into the flask, heat to boiling on a hot
plate,and boil gently for about 20 minutes - the iron powder should
dissolve completely.
Remove from the hot plate and immediately immerse the outlet
end of the glass tubing in saturated sodium bicarbonate solution
contained in a beaker. Cool to room temperature in a water bath
(or ice bath).
Titration;
Remove the stopper, quickly add a few pieces of dry ice, 5 ml
10% potassium iodide solution, and a few ml starch indicator.
Titrate with 0.1N standard potassium iodate solution to a permanent
blue endpoint.
-------
5 Organotin Compounds EPA-1
Calculation:
Calculate the normality of the potassium iodate solution as
follows:
(grams tin standard)
(ml KI03)(.05935)
milliequivalent weight of tin = 0.05935
Calculate the percent tin and organotin compound in the
sample as follows:
Ztln-
-------
October 1975 Ovex EPA-1
Determination of Ovex
by Infrared Spectroscopy
Ovex is the accepted common name for p-chlorophenyl-p-chloro-
benzenesulfonate, a registered acaricide having the chemical
structure:
Molecular formula: C.0H0C1_0_S
Lf. o £3
Molecular weight: 303.2
Melting point: 86.5°C (pure); about 80°C (tech.)
Physical state and color: white crystalline solid (pure), white to
tan flaky solid (technical - about 80 to 90%)
Solubility: practically insoluble in water; moderately soluble in
alcohol; readily soluble in acetone, dichloroethane,
carbon tetrachloride, and aromatic solvents
Stability: chemically stable; hydrolyzed by caustic alkalis; com-
patible with all commonly used spray materials.
(Sometimes imparts an unpleasant taste to fruits because
of chlorophenol which forms on hydrolysis.)
Other names: Ovotran (Dow Chemical), chlorfension (ISO), ovatran
(Argentina), difenson (Denmark), chlorfenizon (France),
ephirsulphonate (USSR), CPCBS, Corotran, Estonmite,
Niagaratran, ovochlor, Sappiran, trichlorfension
-------
2 Ovex EPA-1
This method is primarily for dusts and wettable powders; however,
there is a suggested procedure at the end for emulsifiable concentrates.
Reagents:
1. Ovex standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.2 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, put in 1-2 grams anhydrous sodium sulfate, and
seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50 RPM
on a standard Patterson-Kelley twin shell blender that has
been modified by replacing the blending shell with a box to
hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.08 gram ovex standard into a small glass-stoppered
flask or screw-cap tube, add 10 ml carbon disulfide by pipette,
close tightly,and shake to dissolve. Add a small amount of
anhydrous sodium sulfate to insure dryness. (final cone 8 mg/ml)
-------
3 Ovex EPA-1
Preparation of Sample:
Weigh a portion of sample equivalent to 0.4 gram ovex into
a glass-stoppered flask or screw-cap tube. Add 50 ml carbon
disulfide and 1-2 grams anhydrous sodium sulfate. Close tightly
and shake for one hour. Allow to settle; centrifuge or filter if
necessary, taking precaution to prevent evaporation. (final cone
8 mg ovex/ml)
Determination;
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan the standard and sample from 800 cm
to 740 cm"1 (12.5 p to 13.5 ;j).
Determine the absorbance of standard and sample using the
peak at 770.4 cm (12.98 u) and baseline from 794 cm to
755 cm"1 (12.6 p to 13.25 >i) .
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent ovex as follows:
„ _ (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
The above method is essentially that method (No. 632.0 Nov. 1963)
used by the Pesticide Regulation Division, USDA, now Technical Services
Division, Office of Pesticide Programs, EPA.
A modification of the extraction procedure and a refinement of the
scanning, analytical peak, and baseline wavelengths has been submitted
by the Commonwealth of Virginia, Division of Consolidated Laboratory
Services.
-------
4 Ovex EPA-1
Beltsville Chemical Laboratory suggests the following sample prep-
aration procedure for emulsifiable concentrates:
Weigh a portion of sample equivalent to 0.4 gram ovex
into a small glass-stoppered flask or screw-cap tube, add
50 ml carbon disulfide by pipette, and 1-2 grams anhydrous
sodium sulfate. Close tightly and shake for 10-15 minutes.
Allow to settle. If the carbon disulfide solution is not
clear, add more sodium sulfate and shake again. When the
carbon disulfide solution is sufficiently clear and dry,
proceed with the IR determination. Interfering substances
may or may not be present as shown by a normal or distorted
IR curve.
-------
-------
December 1975 Parathion EPA-1
(Tentative)
Determination of Parathion by
High Pressure Liquid Chromatography
Parathion is the official name for 0,0-diethyl-O-p-nitrophenyl
phosphorothioate, a registered insecticide having the chemical structure:
CH3 CH2 0
Molecular formula: C,_H..NO.PS
10 14 5
Molecular weight: 291.3
Melting/boiling point: ra.p. 6.08C, b.p. 157 to 162°C at 6 mm Hg
Physical state, color, and odor: pale yellow liquid; the technical
product is a brown liquid with a garlic-like odor
Solubility: 24 ppm in water at 25°C; slightly soluble in petroleum oils;
miscible with most organic solvents
Stability: rapidly hydrolyzed in alkaline media (at pH 5 to 6, 1% in
62 days at 25°C); isomerizes on heating to the OS-diethyl
isomer
Other names: ACC 3422, Thiophos (American Cyanamid); E-605, Folidol
Bladan (Bayer); Niran (Monsanto); Fosferno (Plant Protection
Ltd.); thiophos (USSR); Alkron, Aileron, Aphamite, Corothion,
Ethyl Parathion, Etilon, Fosfono, Orthophos, Panthion, Para-
mar, Paraphos, Parathene, Paravet, Phoskil, Rhodiatox,
Soprathion, Strathion
-------
2 Parathion EPA-1
(Tentative)
Reagents;
1. Parathion standard of known % purity
2. Methanol, ACS
Equipment:
1. High pressure liquid chromatograph with UV detector at 254 nm.
If a variable wavelength UV detector is available, other wave-
lengths may be useful to increase sensitivity or eliminate
interference. 235 nm has been found useful for parathion.
2. Suitable column such as:
a. DuPont ODS Pennaphase, 1 meter x 2.1 mm ID
b. Perkin-Elmer ODS Sil-X II-RP, 1/2 meter x 2.6 mm ID
3. High pressure liquid syringe or sample injection loop
4. Usual laboratory glassware
Operat ing Cond it ions;
Mobile phase: 20% methanol + 80% water
Column temperature: 50-558C
Chart speed: 5 min/inch or equivalent
Flow rate: 0.5 to 1.5 ml/min (Perkin-Elmer 1/2 meter column)
Pressure: 700 psi (DuPont 1 meter column)
Attenuation: Adjusted
Conditions may have to be varied by the analyst for the specific
instrument being used, column variations, sample composition, etc. to
obtain optimum response and reproducibility.
-------
3 Parathion EPA-1
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.06 gram parathion standard into a small glass-stoppered
flask or vial, add 20 ml methanol by pipette, dissolve,and mix
well. (final cone 3
Preparation of Sample:
Weigh an amount of sample equivalent to 0.3 gram parathion into
a glass-stoppered flask or vial, add 100 ml methanol by pipette, and
shake thoroughly to dissolve the parathion. With granules or dusts,
shake for 30 minutes on a mechanical shaker or shake by hand inter-
mittently for one hour. Allow any solid matter to settle; filter
or centrifuge if necessary, (final cone 3 ug parathion/ul)
Determination;
Using a high pressure liquid syringe or sample injection loop,
alternately inject three 5 pi portions each of standard and sample
solutions. Measure the peak height or peak area for each peak and
calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the percent
parathion as follows:
„ „ (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA, Beltsville, Md.
-------
December 1975 Parathion EPA-2
(Tentative)
Determination of Parathion by
Gas-Liquid Chromatography
(FID - Internal Standard)
Parathion is the official name for 0,0-diethyl-O-p-nitrophenyl
phosphorothioate, a registered insecticide having the chemical structure;
CH3—CH2—
CH3 CH2 0
Molecular formula: C,_H,,NOCPS
10 14 5
Molecular weight: 291.3
Melting/boiling point: m.p. 6.0°C, b.p. 157 to 162°C at 6 mm Hg
Physical state, color, and odor: pale yellow liquid; the technical
product is a brown liquid with a garlic-like odor
Solubility: 24 ppm in water at 25°C; slightly soluble in petroleum oils;
miscible with most organic solvents
Stability: rapidly hydrolyzed in alkaline media (at pH 5 to 6, 1% in
62 days at 25°C); isomerizes on heating to the OS-diethyl
isomer
Other names: ACC 3422, Thiophos (American Cyanamid); E-605, Folidol
Bladan (Bayer); Niran (Monsanto); Fosferno (Plant Protection
Ltd.); thiophos (USSR); Alkron, Aileron, Aphamite, Corothion,
Ethyl Parathion, Etilon, Fosfono, Orthophos, Panthion, Para-
mar, Paraphos, Parathene, Parawet, Phoskil, Rhodiatox,
Soprathion, Strathion
-------
2 Parathion EPA-2
(Tentative)
Reagents:
1. Parathion standard of known % purity
2. Alachlor standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.2 gram alachlor into a
100 ml volumetric flask and make to volume with acetone.
(cone 2 mg alachlor/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm I.D. glass column packed with 3% QV-17 on
60/80 Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 190°C
Injection temperature: 240°C
Detector temperature: 240°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi (adjusted for specific GC)
Hydrogen pressure: 20 psi (adjusted for specific GC)
Air pressure: 30 psi (adjusted for specific GC)
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Parathion EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.05 gram parathion standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 25 ml of the internal
standard solution and shake to dissolve, (final cone 2 mg parathion
and 2 mg alachlor/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.05 gram parathion
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 25 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the parathion. For
coarse or granular materials, shake mechanically for 30 minutes
or shake by hand intermittently for one hour, (final cone 2 mg
parathion and 2 mg alachlor/ml)
Determination:
Inject 1-2 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2 to
3/4 full scale. The elution order is alachlor, then parathion.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of parathion and alachlor from
both the standard-internal standard solution and the sample-internal
standard solution.
-------
Parathion EPA-2
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
p-p (wt. alachlor)(% purity alachlor)(pk. ht. or area parathipn)
(wt. parathion)(% purity parathion)(pk. ht. or area alachlor)
Determine the percent parathion for each injection of the
sample-internal standard solution as follows and calculate the
average:
y s (wt- alachlor) (% purity alachlor) (pk. ht. or area parathion)-ft007
(wt. sample) (pk. ht. or area alachlor) (RF) (£4~~
*
The following columns also seem satisfactory:
(1) 4' x 2 mm I.D. glass, packed with 5% SE-30 on 80/100 mesh
Chromosorb W HP at 170°C
(2) A1 x 2 mm I.D. glass, packed with 5% OV-210 on 80/100 mesh
Chromosorb W HP at 180°C
This method was submitted by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
Note! This method has been designated as tentative since it is a Va.
Exp. method and because some of the data has been suggested by
EPA's Beltsville Chemistry Lab. Any comments, criticisms,
suggestions, data, etc. concerning this method will be appreciated,
-------
December 1975 Pebulate EPA-1
(Tentative)
Determination of Pebulate by
Gas-Liquid Chromatography (TCD)
Pebulate is the common name for S-propyl butylethylthiocarbamate,
a registered herbicide having the chemical structure*.
—CH3
CH3—CH2—CH2 —S
CH2—CH2—CH2—CH3
Molecular formula: C.JH NOS
Molecular weight: 203.4
Boiling point: 142°C at 21 mm
Physical state, color, and odor: clear yellow liquid with an amine-
like odor
Solubility: 60 ppm in water at 20°C; miscible with acetone, benzene,
ethanol, isopropanol, kerosene, toluene, xylene
Stability: stable; non-corrosive
Other names: Tillam (Stauffer), R-2061
Reagents:
1. Pebulate standard of known £ purity
2. Chloroform, pesticide or spectro grade
-------
2 Pebulate EPA-1
(Tentative)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 6' x 1/4" O.D. aluminum, packed with 20% Dow Silicone
High Vacuum Grease on 60/80 Chromosorb G AW (or
equivalent column - SS or glass would be better)
3. Precision liquid syringe: 10 jjl
4. Usual laboratory glassware
Operating Conditions for TCD:
Column temperature: 160°C
Injection temperature: 185°C
Detector temperature: 185°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas flow rate: adjusted for specific GC .
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
Procedure:
Preparation of Standard:
Weigh 0.5 gram pebulate standard into a 10 ml volumetric flask;
dissolve and make to volume with chloroform, (cone 50 mg/ml)
Preparation of Sample:
For technical material and liquid formulations, weigh a portion
of sample equivalent to 0.5 gram pebulate into a 10 ml volumetric
flask, make to volume with chloroform,and mix thoroughly. (final
cone 50 mg pebulate/ml)
-------
3 Pebulate EPA-1
(Tentative)
For dry formulations, weigh a portion of sample equivalent to
2.5 grams pebulate into a 125 ml screw-cap flask, add by pipette
50 ml chloroform, and shake for one hour. Allow to settle; filter
or centrifuge if necessary, taking precautions to prevent evapora-
tion, (final cone 50 mg pebulate/ml)
Determination:
Using a precision liquid syringe, alternately inject three 5 ^il
portions each of standard and sample solutions. Measure the peak
height or peak area for each peak and calculate the average for both
standard and sample.
Adjustments in attenuation or amount injected may have to be made
to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the percent
pebulate as follows:
7 a (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
This method is based on EPA's Exp. Method No. 50, which was based on
information from Stauffer Chemical Co., Richmond Research Center. Some
of the data has been supplied by EPA's Beltsville Chemistry Laboratory.
Any comments, criticisms, suggestions, data, etc. concerning this method
are welcome.
Note! When operating conditions are such that the retention time of
pebulate is 13.8 minutes, the retention times of known impurities are:
Iso-tillam (iso-pp.bulate) 9.5 min.
N,N'-ethylbutyl n-propyl carbamate 6.8 "
di-n-propyl dithiocarbamate 5.7 "
di-n-propyl disulfide 3.1 "
isopropyl propyl disulfide 2.5 "
ethyl butylamine less than - 1.0 "
n-propyl mercaptan less than - 1.0 "
phosgene less than - 1.0 "
-------
December 1975 Pebulate EPA-2
(Tentative)
Determination of Pebulate by
Gas-Liquid Chromatography
(FID - Internal Standard)
Pebulate is the common name for S-propyl butylethylthiocarbamate,
a registered herbicide having the chemical structure:
0 /CH2—CH3
11 /
CH3—CH2—CH2— S — C—N<^
CH2—CH2—CH2—CH3
Molecular formula: C H NOS
Molecular weight: 203.4
Boiling point: 142°C at 21 mm
Physical state, color, and odor: clear yellow liquid with an amine-like
odor
Solubility: 60 ppm in water at 20°C; iniscible with acetone, benzene,
ethanol, isopropanol, kerosene, toluene, xylene
Stability: stable; non-corrosive
Other names: Tillam (Stauffer), R-2061
Reagents:
1. Pebulate standard of known % purity
2. Cycloate standard of known % purity
-------
2 Pebulate EPA-2
(Tentative)
3. Carbon disulfide, pesticide or spectro grade
4. Chloroform, pesticide or spectro grade
5. Methanol, pesticide or spectro grade
6. Internal Standard solution - weigh 0.2 gram cycloate into a
50 ml volumetric flask, dissolve in, and make to volume with
a solvent mixture consisting of 80% carbon disulfide + 15%
chloroform + 5% methanol. (cone 4 mg cycloate/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 4 mm glass column packed with 3% OV-1 on 60/80 mesh
Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 pi
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 150°C
Injection temperature: 225°C
Detector temperature: 250°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi (adjusted for specific GC)
Hydrogen pressure: 34 psi (adjusted for specific GC)
Air pressure: 28 psi (adjusted for specific GC)
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Pebulate EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.08 gram pebulate standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 20 ml of the internal
standard solution and shake to dissolve. (final cone 4 mg pebulate
and 4 mg cycloate/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.08 gram pebulate into
a small glass-stoppered flask or screw-cap bottle. Add by pipette
20 ml of the internal standard solution. Close tightly and shake
thoroughly to dissolve and extract the pebulate. For coarse or
granular materials, shake mechanically for 30 minutes or shake by
hand intermittently for one hour. (final cone 4 mg pebulate and
4 mg cycloate/ml)
Determination:
Inject 1-2 fil of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2 to
3/4 full scale. The elution order is pebulate, then cycloate.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of pebulate and cycloate from
both the standard-internal standard solution and the sample-internal
standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
-------
Pebulate EPA-2
(Tentative)
(wt. cycloate)(% purity cycloate)(pk. ht. or area pebulate)
(wt. pebulate)(% purity pebulate)(pk. ht. or area cycloate)
Determine the percent pebulate for each injection of the sample
internal standard solution as follows and calculate the average:
_ (wt. cycloate) (% purity cycloate) (pk. ht . or area pebulate)
(wt. sample) (pk. ht. or area cycloate) (RF) //7
<'
Method submitted by Division of Regulatory Services, Kentucky
Agricultural Experiment Station, University of Kentucky, Lexington,
Kentucky 40506.
-------
December 1975 Pebulate EPA-3
(Tentative)
Determination of Pebulate by
Gas-Liquid Chromatography
(FID - Internal Standard)
Pebulate is the common name for S-propyl butylethylthiocarbamate,
a registered herbicide having the chemical structure:
0 XH2 —CH3
" /
CH3—CH2—CH2 — S— C — N<"
CH2—CH2 —CH2 —CH3
Molecular formula: C H NOS
Molecular weight: 203.4
Boiling point: 1A2°C at 21 mm
Physical state, color, and odor: clear yellow liquid with an amine-like
odor
Solubility: 60 ppm in water at 20°C; miscible with acetone, benzene,
ethanol, isopropanol, kerosene, toluene, xylene
Stability: stable; non-corrosive
Other names: Tillatn (Stauffer) , R-2061
Reagents;
1. Pebulate standard of known % purity
2. S-Ethyl dipropylthiocarbamate (EPIC) standard of known % purity
-------
Pebulate EPA-3
(Tentative)
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.1 gram EPTC into a 50 ml
volumetric flask; dissolve in and make to volume with acetone.
(cone 2 mg EPTC/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm glass column packed with 5% SE-30 on 80/100 mesh
Chroraosorb W HP (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 130°C
Injection temperature: 180°C
Detector temperature: 180°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi (adjusted for specific GC)
Hydrogen pressure: 20 psi (adjusted for specific GC)
Air pressure: 30 psi (adjusted for specific GC)
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Pebulate EPA-3
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.06 gram pebulate standard into a small glass-stoppered
flask or screw-cap bottle. Add bv pipette 20 ml of the internal
standard solution and shake to dissolve. (final cone 3 mg pebulate
and 2 mg EPTC/ml)
Preparation of Sample:
Weigh a portion of sample eouivalent to 0.06 gram pebulate into
a small glass-stoppered flask or screw-cap bottle. Add by pipette
20 ml of the internal standard solution. Close tightly and shake
thoroughly to dissolve and extract the pebulate. For coarse or
granular materials, shake mechanically for 30 minutes or shake by
hand intermittently for one hour. (final cone 3 mg pebulate and
2 mg EPTC/ml)
Determination:
Inject 1-2 ^il of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2 to
3/4 full scale. The elution order is EPTC, then pebulate.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of pebulate and EPTC from both
the standard-internal standard solution and the sample-internal
standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
-------
Pebulate EPA-3
(Tentative)
(wt. EPTC) (% purity _EPTC)_(2k_1_ ht. or area pebulate)
RF — ~r ~ '/ -•
(wt. oebulate)(% purity pebulate)(pk. ht. or area EPTC)
Determine the percent pebulate for each injection of the
sample-internal standard solution as follows and calculate the
average:
_ Cwt • EPTC)_(_%_purity EPTC)jpk.__ht_. or area pebulate)J4j»^
0 ~ (wt. sample) (pk. ht. or area EPTC) (RF) / \J-\j
Method submitted by the Commonwealth of Virginia, Division of Laboratory
Services, 1 North 14th Street, Richmond, Virginia 23219.
Note! This method has been designated as tentative since it is a Va.
Exp. method and because some of the data has been suggested by
EPA's Beltsville Chemistry Laboratory. Any comments, criticisms,
suggestions, data, etc. concerning this method will be appreciated,
-------
March 1976
Phenols and Chlorophenols EPA-1
Definition, Structure, and Technical Data
This group of compounds consists of various aliphatic, aromatic,
and chlorine substituted phenols.
These compounds are registered as germicides and/or fungicides.
Many are used in the form of alkali salts or amine salts.
The following physical and chemical data are for the free phenols.
o-phenylphenol
C12H10°
mol. wt. 170.20
white flaky crystals; mild characteristic odor; mo 55.5-57.5°C:
bp 280-284°C; practically insoluble in water, soluble in alkali
hydroxide solutions and most organic solvents. Other names:
Dowcide 1, o-hydrodiphenyl, orthoxenol
g-tert-butylphenol C H ,0 mol. wt. 150.21
-------
2 Phenols and Chlorophenols EPA-1
white crystalline solid; distinctive odor; mp 98-100°C; bp 237-239°C;
practically insoluble in cold water, soluble in alcohol, ether.
Other names: Butylphen
p-tert-amylphenol C H 0 mol. wt. 164.24
OH
0 I
CH3
white crystals (irritating to skin); mp 94-95°C; bp 262.5°C;
practically insoluble in water, soluble in alcohol, ether, benzene,
chloroform. Other names: p-tert-pentylphenol, Pentaphen
pentachlorophenol C Cl OH mol. wt. 266.35
OH
Cl
Cl
white powder or crystals; very pungent odor when hot; mp 190-191°C;
bp about 309-310°C with decomposition; almost insoluble in water,
soluble in dilute alkali, alcohol, acetone, ether, pine oil, benzene;
slightly soluble in cold petroleum ether. Other names: PCP, Penta,
Santophen 20
-------
Phenols and Chlorophenols EPA-1
4-chloro-3,5-xylenol C0H0C10 mol. wt. 156.61
. . __ -
crystals with phenolic odor; mp 115.5°C; bp 246°C: volatile with steam;
one gram dissolves in 3 liters of water at 20°C; more stable in hot
water; soluble in 1 part of 95% alcohol; soluble in ether, benzene,
terpenes, fixed oils, and solutions of alkali hydroxides. Other names:
p-chloro-m-xylenol, Benzytol, 4-chloro-3,5-dimethylphenol, 2-chloro-
5—hydroxy-l,3-dimethylbenzene
o-benzyl-p-chlorophenol C H CIO mol. wt. 218.69
OH
rCH2
Cl
white to light tan or pink flakes; slight phenolic odor; mp 48.5°C;
insoluble in water; highly soluble in alcohol and other organic solvents;
dispersible in aqueous media with the aid of soaps or synthetic dispersing
agents; non-corrosive to most metals. Other names: Santophen 1, Septiphene,
Clorophene, 2-benzyl-4-chlorophenol
-------
Phenols and Chlorophenols EPA-1
4-chloro-2-phenylphenol
OH
r
Cl
6-chloro-2-phenylphenol
OH
raol. wt. 204.65
clear colorless to straw-colored viscous liquid
with faint characteristic odor
boiling range 5-95% 146-158.7°C (5 mm)
readily soluble in most organic liquids
composition 80% 4-chloro-2-phenylphenol
20% 6-chloro-2-phenylphenol
Other phenols and chlorinated phenols not listed here are also
used as germicides or fungicides and may be found in various commercial
formulations, such as:
4-Chloro-2-cyclopentylphenol
2,2'-Methylenebis (4-chlorophenol)
2,2'-Methylenebis (3,4,6-trichlorophenol)
-------
March 1976 Phenols and Chlorophenols F.PA-2
Determination of o-Phenylphenol in Disinfectant
Formulations by Ultraviolet Spectroscopv
For definition, structure, and technical data on o-phenylphenol ,
see Phenols and Chlorophenols EPA-1 .
This method is intended primarily for alcohol solutions of ahout
0.1% o-phenylphenol (tert-amylphenol) interferes very little). Data and
information on the use of this method for other phenols and chloro-
phenols will be appreciated by the editorial committee.
Reagents :
1. o-phenylphenol standard of known 7, purity
2. Sodium hydroxide, IN aqueous solution
3. Ethanol, ACS
4. Hexane, purified. Extract 250 ml n-hexane with two 20 ml
Dortions of IN sodium hydroxide solution and one 20 ml
portion of water; discard the extracts.
5. Sulfuric acid, 1+4 solution
Equipment :
1. Ultraviolet spectrophotometer , double beam ratio recording
with matched 1 cm cells
2. Rotary evaporator
3. Usual laboratory glassware
Procedure:
of Standard :
Weigh 0.04 gram o-phenylphenol standard into a 100 ml volumetric
flask, add 5 ml IK sodium hydroxide solution, dissolve and dilute
-------
2 Phenols and Chlorophenols EPA-2
to volume with water. Mix thoroughly and pipette 2 ml into a
50 ml volumetric flask. Add 5 ml IN sodium hydroxide solution,
25 ml ethanol, dilute to volume with water, and mix thoroughly.
(final cone 16 jig o-phenylphenol/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.008 gram o-phenvl-
phenol into a 125 ml Erlenmeyer flask, add 1 drop of IX sodium
hydroxide, and evaporate to dryness on a rotary evaporator.
Dissolve the residue in about 40 ml water and 20 ml hexane,
transfer quantitatively to a 250 ml separatory funnel, add 5 ml
IN sodium hydroxide solution, shake, and allow the layers to
separate.
Transfer the aqueous layer to a second 250 ml separatory
funnel. Wash the hexane layer in the first senaratory funnel
with two 20 ml portions of water and add the washings to the
second separatory funnel. Acidify with 3 ml of 1+4 sulfuric
acid solution and extract with 50 ml hexane. Repeat with 25 ml
hexane and combine the hexane extracts in a 125 ml separatory
funnel. Extract the combined hexane layers with 20 ml IN sodium
hydroxide solution; transfer the alkaline aqueous extract into
a 100 ml volumetric flask. Extract the hexane with 20 ml water
and add to the 100 ml volumetric flask. Dilute to volume with
water and mix thoroughly.
Pipette 10 ml of this solution into a 5n ml volumetric flask,
add 3 ml IN sodium hydroxide solution and 25 ml ethanol, dilute to
volume with water, and mix thoroughly. (final cone 16 ug o-phenyl-
phenol/ml)
-------
Phenols and Chlorophenols EPA-2
UV _D_e_termina_t_ion:
With the UV spectrophotomcter at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 312 nm with 0.1N
sodium hydroxide solution in each cell. Scan both the standard
and sample from 360 nm to 260 nm with O.IK sodium hydroxide
solution in the reference cell. Measure the absorbance of both
standard and sample at 312 nm.
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent o-phenylphenol as follows:
„ _ _(_absj._ s ample)(cone, std in_jug/ml) (_% purity std)
(abs. std)(conc. sample in
-------
March 1976 Phenols and Chlorophenols EPA-3
Determination of Chlorophenols by the
Total Chloride Lime Fusion Method
For definition, structure, and technical data on Chlorophenols,
see Phenols and Chlorophenols EPA-1.
Principle of the Method:
The method was developed primarily for pentachlorophenol in oil
solution';; however, it may be used for other chlorinated phenols. It
is not applicable to samples containing other halogens unless these
halogens can be determined and appropriate corrections made. This
method is based on destroying organic material by heating and absorb-
ing the liberated hydrochloric acid in calcium hydroxide. The
hydroxide is neutralized with nitric acid and the chloride determined
potentiometrically. Organic matter should be limited to 2 grams and
the chlorinated phenol to35mg in terms of chlorine.
Reagents;
1. Fusion mixture - 9 parts calcium hydroxide powder plus 1 part
potassium nitrate powder, thoroughly mixed. (see note below)
2. Nitric acid, concentrated, ACS
3. Silver nitrate, 0.1N standardized solution
Note: All reagents should be virtually chloride-free. ACS speci-
fications should meet this requirement; however, for greatest
accuracy a blank on all reagents should be run using the
same amounts and method as for the sample.
4. Phenolphthalein indicator solution
-------
Phenols and Chlorophenols EPA-3
Equipment:
1. Potentiometric titrimeter equipped with a glass reference
electrode and a silver electrode
2. Iron crucible, 100 ml capacity, with cover
3. Meker burner, adjustable from minimum air to maximum air
4. Tripod stand and metal triangle (to hold crucible)
5. Ice bath
6. Magnetic stirrer
7. Usual laboratory glassware
Determination:
Place 100 grams of the calcium hydroxide-potassium nitrate mixture
in a 100 ml iron crucible, tap gently to settle the contents, and form
a small depression with the round bottom of a test tube. From a weigh-
ing burette add a weight of sample equivalent to 0.035 gram chlorine.
(Solid samples may be mixed with a little fusion mixture and placed in
the depression.)
Place 20 grams of the fusion mixture over the sample in the crucible
and tap gently on a hard surface to settle and evenly distribute the
fusion mixture. It is essential that the fusion mixture be uniformly
packed (settled) so that no air pockets are present and thorough and
even heating results.
The crucible with cover (to suppress burning of volatile materials
on the surface of the fusion mixture) should be placed in a metal
triangle on a ring stand so that the bottom is one-half inch above the
top of a Meker burner. With the air supply almost completely shut off
-------
3 Phenols and Chlorophenols EPA-3
and using a very luminous flame, the crucible is heated for about 15
minutes, allowing the flame to completely engulf the crucible all
around. Gradually increase the flame temperature (increase the air)
to maximum over the next ten minutes until the bottom of the crucible
is red hot. Heat at full heat for at least 30 minutes. Samples
should be free of unburned carbon; however, a small amount usually
presents no errors. Surface should be free of large cracks.
Cool the crucible until it can be handled, then empty the contents
into a 600 ml beaker, scraping any adhering fusion mixture into the
beaker. Cautiously add about 100 ml water to the beaker and rinse the
crucible with small portions of water into the beaker. Place the
beaker in an ice bath in a glass dish on a magnetic stirrer. Put a
glass or teflon-coated stirring bar and a few drops of phenolphthalein
in the beaker and cover with a watchglass. While stirring, cautiously
and slowly pour sufficient cone, nitric acid (50 to 60 ml) slowly down
the side of the beaker to neutralize the sodium carbonate (keep the
beaker covered as much as possible with the watchglass). Cool, and
determine the chloride content potentiometrically, titrating with 0.1N
silver nitrate solution. A blank should be run on each new batch of
fusion mixture and with each change of reagent. Corrections in calcu-
lation should be made (usually about 0.05-0.06 ml silver nitrate
solution subtracted from the ml silver nitrate used for the sample
titration).
-------
Phenols and Chlorophenols EPA-3
Calculations:
Calculate the percent chlorine and chlorinated phenol as follows:
(net ml AgNO )(N AgNO )(0.03545)(100)
% chlorine = ,— —:
(gram sample)
(0.03545 = milliequivalent weight of chlorine)
% Chlorinated phenol = % chlorine X factor Cl to chlorinated phenol
-------
March 1976 Phenols and Chlorophenols EPA-4
Determination of o-Phenylphenol and Sodium Salt
of o-Phenylphenol by Bromination and Titration
For definition, structure, and technical data on o-phenylphenol,
see Phenols and Chlorophenols EPA-1.
Principle of the Method:
Sodium o-phenylphenol formulations are dissolved in water and
filtered. o-Phenylphenol in oil formulations is distilled from acid
solution, made alkaline, and evaporated to remove volatile organic
interfering substances. A known portion of prepared sample is reacted
with excess bromate-bromide solution and the excess determined iodo-
metrically using standard sodium thiosulfate. The o-phenylphenol is
calculated from the net difference in sodium thiosulfate used by a
blank and by the sample.
Reagents;
1. Sodium hydroxide solution, 10% aqueous solution
2. Hydrochloric acid, concentrated, ACS
3. Bromate-bromide 0.1N solution - dissolve 2.78 grams of potassium
bromate and 15 grams potassium bromide in water and make to one
liter. This solution need not be standardized if a blank using
the same quantity as the sample is run each time.
A. Potassium iodide, 40% aqueous solution
5. Sodium thiosulfate, 0.1N standardized solution
6. Starch indicator solution - 1 gram soluble starch boiled 2
minutes in 100 ml water
-------
2 Phenols and Chlorophenols EPA-4
Equipment:
1. One liter distilling flask with condenser
2. Filtration apparatus
3. Hot plate
4,. Air stream
5. 500 ml iodine flask
6. Titration apparatus
7. Usual laboratory glassware
Procedure;
Preparation of Sample:
(a) o-phenylphenol in oil solutions - weigh a portion of sample
equivalent to 0.04 gram o-phenylphenol into a one liter distilling
flask, add 10 ml of 10% sodium hydroxide solution, and dilute to
about 600 ml. Add 20 ml concentrated hydrochloric acid and a few
boiling chips, and distill about 400 ml into a 1000 ml Erlenmeyer
flask. Interrupt the distillation, add about 400 ml water to the
distilling flask, and distill an additional 300 ml into the same
1000 ml Erlenmeyer flask. Add 15 ml of 10% sodium hydroxide solution
to the distillate and boil down to about 50 ml using a stream of air
against the surface of the liquid to prevent frothing. Transfer
quantitatively to a 500 ml iodine flask.
(b) Sodium salt of o-phenylphenol - weigh a portion of sample
equivalent to 1 gram sodium salt of o-phenylphenol into a 200 ml
volumetric flask; dissolve in and make to volume with water. Filter,
discarding the first 50 ml, pipette a 10 ml portion of the clear
filtrate into a 500 ml iodine flask, and add 50 ml water.
-------
Phenols and Chlorophenols EPA-4
Titration:
Pipette 25 ml 0.1N bromate-bromide solution into the iodine
flask, add 15 ml concentrated hydrochloric acid, stopper, and
allow to stand 15 minutes in a dark place with occasional shaking.
Remove the stopper just sufficiently to quickly add 5 ml of 40%
potassium iodine solution, taking care that no bromine vapor
escapes. Restopper at once. ShaVe thoroughly. Remove the stopper,
rinsing it and the well of the flask with water so that the washings
flow into the flask. Wash down the inside walls of the flask with
5-10 ml water.
Titrate the liberated iodine with 0.1N standard thiosulfate
solution using starch indicator near the endooint.
Run a blank in the same way using the same quantity of reagents
beginning with 50 ml of water in an iodine flask.
Calculations ;
Subtract the ml used for the sample titration from the ml used
for the blank titration to obtain the net ml equivalent to the
o-phenylphenol in the sample.
Calculate the o-phenylphenol in oil solution as under sample
preparation (a) as follows:
(net ml Na S.O,)(N Na_S,0.) (0.04255) (100)
£. 4. J £ Z O
(grams sample)
0.04255 - milliequivalent weight of o-phenylphenol
Calculate the sodium salt of o-phenylphenol as under sample
preparation (b) as follows:
(net ml Na^OjMN Na2S203) (0.04805) (100)
(grams sample) (10/200)
0.04805 - milliequivalent weight of the sodium salt of
o-phenylphenol
10/200 - dilution factor in sample preparation
-------
Phenols and Chlorophenols EPA-4
Reactions:
1. Release of bromine from bromate-bromide solution:
KBrO + 6KBr + 6HC1
+ KBr + 6KC1 4- 3H 0
2. Bromination of the o-phenylphenol:
ONa
ONa
2HBr
3. Release of iodine from excess bromine:
(excess)Br + 2KI
+ 2KBr
4. Titration of iodine with sodium thiosulfate:
V
•2NaI + Na.S.O,
246
-------
March 1976 Phenols and Chlorophenols EPA-5
( Tentative)
Determination of Pentachlorophenol
by High Pressure Liquid Chromatography
For definition, structure, and technical data on pentachlorophenol,
see Phenols and Chlorophenols EPA-1.
Reagents;
1. Pentachlorophenol standard of known % purity
2. Benzyl benzoate standard of known % purity
3. Ethanol, ACS
4. Internal standard solution - weigh 5 grams benzyl benzoate
into a 50 ml volumetric flask; dissolve in and make to
volume with ethanol, (cone 100 mg/ml)
Equipment;
1. High pressure liquid chromatograph with UV detector at 254 nm.
If a variable wavelength UV detector is available, other
wavelengths may be useful to increase sensitivity or eliminate
interference.
2. Column: 0.5 meter x 2.1 mm ID packed with DuPont ODS Perma-
phase (or equivalent column)
3. High pressure liquid syringe or sample injection loop
4. Usual laboratory glassware
Operating Conditions;
Mobile phase: 30% methanol + 70% water
Column temperature: 60CC
Chart speed: 12 inches/minute
Flow rate: 1.0 ml/minute
-------
Phenols and Chlorophenols EPA-5
(Tentative)
Pressure: 800-1000 psi
Detector: UV at 254 nm
Attenuation: adjusted
Conditions may have to be varied by the analyst for the specific
instrument being used, column variations, sample composition, etc. to
obtain optimum response and reproducibility.
Procedure:
Preparation of Standard;
Weigh 0.05 gram pentachlorophenol standard into a 100 ml
volumetric flask, add 5 ml of the internal standard solution by
pipette, make to volume with ethanol, and mix thoroughly. (cone
0.5 rag pentachlorophenol and 5 mg benzyl benzoate/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.05 g pentachloro-
phenol into a 100 ml volumetric flask, add 5 ml internal standard
solution by pipette, make to volume with ethanol, and mix thor-
oughly, (cone 0.5 mg pentachlorophenol and 5 mg benzyl benzoate/ml)
Determination;
Inject 5 pi of standard solution and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is pentachlorophenol, then
benzyl benzoate. Proceed with the determination, making at least
three injections each of standard and sample solutions in random
order.
-------
Phenols and Chlorophenols EPA-5
(Tentative)
Calculation;
Measure the peak heights or areas of pentachlorophenol and
benzyl benzoate from both the standard-internal standard solution
and the sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
IS = internal standard = benzyl benzoate
PCP = pentachlorophenol
(wt. IS)(% purity IS)(pk. ht. or area PCP)
(wt. PCP)(% purity PCP)(pk. ht. or area IS)
Determine the percent PCP for each injection of the sample-
internal standard solution as follows and calculate the average:
7 = (wt. IS)(% purity IS)(pk. ht. or area PCP)(100)
(wt. sample)(pk. ht. or area 1S)(RF)
Method submitted by Elmer H. Hayes, EPA, Beltsville, Md.
-------
March 1976 Phenols and Chlorophenols EPA-6
(Tentative)
Determination of Phenols and Chlorophenols
by Gas-Liquid Chromatography (FID)
For definition, structure, and technical data on these compounds,
see Phenols and Chlorophenols EPA-1.
This method has been found suitable for o-phenylphenol, p-tert-
amyl-phenol, and o-benzyl-p-chlorophenol; however, with modification
it should be suitable for several other phenol and chlorophenol compounds.
Reagents;
1. Phenol or chlorophenol standard of known % purity
2. Acetone, ACS
3. Ethyl ether, ACS
4. Sulfuric acid, ACS, 1+9 solution
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 1/4" glass column packed with 5% XE-60 on
60/80 Chromosorb W DMCS (or equivalent column)
3. Precision liquid syringe: 10 ul
4. 125 ml separatory funnels
5. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: See under determination
Injection temperature: 250°C
Detector temperature: 250°C
-------
Phenols and Chlorophenols EPA-6
(Tentative)
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi, adjusted for particular GC
Hydrogen pressure: 20 psi, adjusted for particular GC
Air pressure: 30 psi, adjusted for particular GC
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
Procedure:
Preparation of Standard:
Weigh 0.04 gram o-phenylphenol or p-tert-amylphenol, or 0.08
gram o-benzyl-p-chlorophenol into a 25 ml volumetric flask,
dissolve in, and make to volume with acetone. Mix well. (cone
1.6 mg/ml each of o-phenylphenol and p-tert-amylphenol and 3.2
mg/ml of o-benzyl-p-chlorophenol)
(Other phenols may require slightly different concentrations.)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.04 gram o-phenyl-
phenol or p-tert-amylphenol, or 0.08 gram o-benzyl-p-chlorophenol
into a 125 ml separatory funnel. Make slightly acidic with 1+9
sulfuric acid; then add 10 ml in excess. Extract three times with
25-50 ml portions of ethyl ether,collecting the extracts in a
second separatory funnel. Wash once with a few ml of 1+9 sulfuric
acid. Drain the ether extracts into a beaker, rinsing the separa-
tory funnel with a few ml ether twice and adding the washings to
the beaker. Allow the ether to evaporate (overnight) at room
temperature using no heat or air jet. Dissolve the residue in a
-------
Phenols and Chlorophenols EPA-6
(Tentative)
small amount of acetone, quantitatively transfer to a 25 ml volu-
metric flask, and make to volume with acetone. (Samples in
aerosols usually do not require extraction and can be weighed
directly into a volumetric flask and made to volume.)
(If only one phenol is present, concentration after the above
procedure should be 1.6 rag/ml each for o-phenylphenol and p-tert-
amylphenol and 3.2 mg/ral for o-benzyl-p-chlorophenol.)
Determination:
A column temperature of 180°C is sufficient for o-phenylphenol
and p-tert-amylphenol, eluting in that order. However, a 220°C
temperature is needed for o-benzyl-p-chlorophenol to prevent an
excessively long retention time.
Using a precision liquid syringe, alternately inject three
5 ul portions each of standard and sample solutions. Measure the
peak height or peak area for each peak and calculate the average
for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
When several phenols or chlorophenols are present in the same
sample, a standard approximating the sample composition should be
made. In this case the column temperature may have to be programmed
from about 150°C to 250°C.
Calculation:
From the average peak height or peak area calculate the percent
phenol or chlorophenol compound as follows:
7 _ (pk. ht. or area sample)(wt. std injected)(% purity of std)
'° """ / 1 i . . * * \ s . t . • . i \
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA, OPP, TSD, Beltsville, Md.
-------
March 1976 Phenols and Chlorophenols EPA-7
(tentative)
Determination of 4-Chloro-3,5-Xylenol
by Gas-Liquid Chromatography (TCD and/or FID)
For definition, structure, and technical data on 4-chloro-3,5-
xylenol, see Phenols and Chlorophenols EPA-1.
Reagents:
1. 4-chloro-3,5-xylenol standard of known % purity
2. Acetone, ACS
3. Petroleum ether, ACS
4. Ethyl ether, ACS
5, Sodium hydroxide, IN aqueous solution
6. Sulfuric acid, 1+4 solution
Equipment:
1. Gas-Liquid Chromatograph with thermal conductivity detector
(TCD) or flame ionization detector (FID)
2. Column for TCD: 5' x 1/4" O.D. glass column packed with
20% SE-30 on 60/80 Chromosorb, AW, DMCS (or equivalent column)
3. Column for FID: 6' x 1/4" O.D. glass column packed with
3% OV-1 on 80/100 Gas Chrom Q (or equivalent column)
4. Precision liquid syringe: 10 jal or 50/il
5. Usual laboratory glassware
Procedure using Thermal Conductivity Detector:
Operating Conditions for TCD:
Column temperature: 210°C
Injection temperature: 240°C
-------
Phenols and Chlorophenols EPA-7
(Tentative)
Detector temperature: 270°C
Carrier gas: Helium
Flow rate: 100 ml/min
Operating conditions for filament current, column temperature,
or gas flow should be adjusted by the analyst to obtain optimum
response and reproducibility.
Preparation of Standard:
Weigh 0.2 grams 4-chloro-3,5-xylenol standard into a small
glass-stoppered flask or screw-cap bottle, add by pipette 10 ml
acetone, and shake to dissolve. (cone 20 mg/ml)
Preparation of Sample:
(a) For technical material, weigh a portion of sample equiv-
alent to 0.2 gram 4-chloro-3,5-xylenol into a small glass-stoppered
flask or screw-cap bottle, add by pipette 10 ml acetone, and shake
to dissolve. (cone 20 mg/ml)
(b) For low % formulations in oils, weigh a portion of sample
equivalent to 0.2 gram 4-chloro-3,5-xylenol into a 250 ml separa-
tory funnel. Add about 100 ml petroleum ether and extract three
times with 25 ml IN sodium hydroxide solution. Combine the extracts
into a second 250 ml separatory funnel, acidify with 1+4 sulfuric
acid solution, and add 5 ml in excess. Extract twice with 75 ml
ethyl ether. Filter the ether extracts through a cotton plug into
a 300 ml flask and evaporate almost to dryness on a steam bath,
allowing the last traces of ether to evaporate spontaneously from
the warm flask. Dissolve the residue, transfer quantitatively to
a 10 ml volumetric flask, and make to volume with acetone. Mix
well. (cone 20 mg/ml)
-------
Phenols and Chlorophenols EPA-7
(Tentative)
Determination:
Using a precision liquid syringe, alternately inject three
30-40 ul portions each of standard and sample solutions. Measure
the peak height or peak area for each peak and calculate the
average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the
percent 4-chloro-3,5-xylenol as follows:
7 a (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Procedure for Flame lonization Detector:
Operating Conditions for FID:
Column temperature: 145°C
Injection temperature: 225°C
Detector temperature: 220°C
Carrier gas: Nitrogen (30 ml/min)
Carrier gas pressure: 60 psi, adjusted for particular GC
Hydrogen pressure: 20 psi, adjusted for particular GC
Air pressure: 30 psi, adjusted for particular GC
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Phenols and Chlorophenols EPA-7
(Tentative)
Preparation of Standard;
Same as for TCD except use a 100 ml volumetric flask instead
of a 10 ml volumetric flask. (cone 2 rag/ml)
Preparation gf Sample:
Same as for TCD except use a 100 ml volumetric flask instead
of a 10 ml volumetric flask. (cone 2 mg/ml)
Determination:
Using a precision liquid syringe, alternately inject three
2-4 ^il portions each of standard and sample solutions. Measure
the peak height or peak area for each peak and calculate the
average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent 4-chloro-3,5-xylenol as follows:
7 - (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Eva Santos and Dean Hill, EPA Region IX, San
Francisco, California.
-------
April 1976 Phenols and Chlorophenols EPA-8
(Tentative)
Determination of Phenols and Chlorophenols by
Gas-Liquid Chromatography (TCD-IS-BSA derivatization)
For definition, structure, and technical data on these compounds,
see Phenols and Chlorophenols EPA-1.
Principle of the Method:
Trimethyl silyl derivatives of phenols and Chlorophenols yield
sharp, symmetrical peaks ideal for quantitative measurement. These
peaks are also stronger and thus increase the sensitivity of the
analysis. The BSA reagent produces no interference.
The precision of this method is very good — the same sample
analyzed several times was found to give almost identical results.
Also, the stability of the BSA derivative gave no detectable change
over six days. Germicide formulations containing such compounds as
soaps, triethanolamines, oils, and other active and inert ingredients
seemed to present no problems and the results obtained were satisfactory.
A portion of prepared sample solution in chloroform is evaporated
to dryness, treated with BSA reagent, has a portion of internal standard
solution added, and is chromatographed with good results.
Reagents:
1. Phenol or chlorophenol standards of known % purity (see table
of components and internal standards)
2. Internal standards, technical (or better) (see table of components
and internal standards)
3. Chloroform, ACS
-------
Phenols and Chlorophenols EPA-8
( Tentative)
4. N,0-bis(trimethylsilyl)acetamide (BSA)
5. Sodium hydroxide, IN solution
6. Sulfuric acid, 10% solution
7. Ethyl ether, ACS
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 4' x 1/4" O.D. glass packed with 4% SE-30 80/100 mesh
Diatoport S (or equivalent column)
3. Precision liquid syringe: 10 pi
4. Rotary evaporator
5. Steam bath with gentle stream of air
6. Usual laboratory glassware
Operating Conditions for TCD:
Column temperature: 165°C
Injection temperature: 215°C
Detector temperature: 230°C
Filament current: 200 ma
Carrier gas: Helium 25 ml/min
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Phenols and Chlorophenols EPA-8
(Tentative)
Procedure:
Preparation of Standards;
Weigh 0.25 gram of the phenol or chlorophenol standard into
a 25 ml volumetric flask; dissolve in and make to volume with
chloroform. (cone 10 rag/ml)
Preparation of Samples:
(a) For samples containing an appreciable amount of alcohol,
weigh a portion of sample equivalent to 0.5 gram phenol or
chlorophenol into a standard taper Erlenmeyer flask, make
alkaline with IN sodium hydroxide solution, and evaporate the
water and alcohol to about 3-4 ml. Transfer quantitatively with
50 ml water into a 250 ml separatory funnel, neutralize with 10%
sulfuric acid solution, and add 10 ml in excess. Extract two
times with 50 ml of ethyl ether. Combine the ether extracts and
wash with 10 ml water. Filter and dry the ether extracts by
passing thru a plug of cotton and anhydrous sodium sulfate into
a 250 ml standard taper Erlenmeyer flask and evaporate on a
rotary evaporator (the ether may also be evaporated with a stream
of dry air). Quantitatively transfer to a 50 ml volumetric flask
and make to volume with chloroform. Mix thoroughly.
(b) For samples containing slight amounts (3%) of alcohol,
weigh a portion of sample equivalent to 0.5 gram phenol or chloro-
phenol directly into the 250 ml separatory funnel and proceed as
above beginning "neutralize with 10% sulfuric acid ..."
Preparation of Internal Standard Solutions;
Prepare chloroform solutions of the internal standard solutions
as follows:
(1) n-tetradecane, 1 gram in 50 ml, cone 20 mg/ml
(2) lindane, 4 grams in 50 ml, cone 80 mg/ml
-------
Phenols and Chlorophenols EPA-8
(Tentative)
(3) n-hexadecane, 0.5 gram in 50 ml, cone 10 mg/ml
(4) benzylbenzoate, 0.625 gram in 50 ml, cone 12.5 mg/ml
(5) di-2-ethylhexylphthalate(A), 0.5 gram in 50 ml, cone 10 mg/ml
(6) di-2-ethylhexylphthalate(B), 2 grams in 50 ml, cone 40 mg/ml
These concentrations are suggested for a 1:1 peak height ratio
with 20 mg of phenol or chlorophenol.
De t ermina t ion:
Pipette a 2 ml aliquot of the standard and sample solutions
into separate 15 ml screw-cap vials and evaporate the chloroform
to near dryness with a gentle stream of dry air. Add 1 ml BSA
reagent, close tightly, shake to dissolve the residue, and allow
to stand 10 minutes with occasional shaking. Add 1 ml of the
appropriate internal standard as listed in the table below and
mix well.
2 ml phenol compound (20 mg)
p-tert-butylphenol
p-tert-amylphenol
o-phenylphenol
4-chloro-2-cyclopentylphenol
o-benzyl-p-chlorophenol
4 or 6-chloro-2-phenylphenol
2,2'-methylenebis(3,4,6-trichlorophenol)
2,2'-methylenebis(4-chlorophenol)
1 ml internal standard
(mg as listed)
n-tetradecane 20 mg
n-tetradecane 20 mg
lindane 80 mg
n-hexane 10 mg
benzyl benzoate 12.5 mg
benzyl benzoate 12.5 mg
di-2-ethylhexylphthalate(A) 10 mg
10 mg
di-2-ethylhexylphthalate(B) 40 mg
40 mg
-------
Phenols and Chlorophenols EPA-8
(Tentative)
Adjust the GC parameters and the size of the injection (3-4
so that an injection of ether solution shows complete separation
of the internal standard and the derivatized phenol compound within
10 minutes and so that there is no interference by other peaks.
The height of both peaks should be between 1/2 to 3/4 full scale.
Make at least three injections of the standard solution. The
ratio of the peak height of the derivatized standard to the peak
height of the internal standard should be within 3% of the ratio
of the previous injection. This will indicate that the instrument
has reached equilibrium and that the operator has standardized his
injection technique.
Proceed, making at least three injections of each solution,
alloxfing time for any accompanying peak in the sample to elute
before making the next injection.
Calculation:
Measure the peak heights of the internal standard and the
derivatized standard phenol. Determine the RF value for each
injection and average.
., ^ (wt. internal std.)(peak ht. of derivatized phenol)
(wt. phenol std.)(% purity phenol std.)(peak ht. internal std.)
Measure the peak heights of the internal standard and the
derivatized sample phenol and calculate the average. Determine the
percent phenol as follows:
(wt. internal std.) (peak ht . of derivatized
" ~ (wt. sample) (peak height of internal standard) (RF)
-------
December 1975
Phenothiazine EPA-1
(Tentative)
Determination of Phenothiazine
by Infrared Spectroscopy
Phenothiazine is a registered oral insecticide and anthelmintic
having the chemical structure:
H
Molecular formula: C H.NS
Molecular weight: 199.3
Melting point: 185°C, sublimes 130°C (1 mm); b.p. 371°C
Physical state, color, odor, and taste: tasteless, crystalline solid
with a slight odor; almost colorless when freshly
sublimed, darkens to deep olive-green on exposure
to light
Solubility: insoluble in water, chloroform; slightly soluble in alcohol,
ether; soluble in acetone, benzene
Stability: oxidized in the presence of air and light to phenothiazone
and thionol
Other names: thiodiphenylamine
Reagents;
1. Phenothiazine standard of known % purity
2. Benzene, pesticide or spectro grade
3. Carbon disulfide, pesticide or spectro grade
4. Sodium sulfate, anhydrous, granular
-------
Phenothiazine EPA-1
(Tentative)
Equipment;
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.5 mm NaCl or KBr cells
2. Soxhlet extraction apparatus
3. Steam bath
A. Compressed air source
5. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.15 gram phenothiazine standard into a small glass-
stoppered flask or screw-cap tube, add by pipette 25 ml carbon
disulfide, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 6 mg/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 1.5 grams phenothiazine
into a Soxhlet thimble, plug with cotton or glass wool, and extract
with benzene 3-4 hours. Cool, transfer to a 250 ml volumetric
flask, and make to volume with benzene. Evaporate a 25 ml aliquot
to just dryness using a gentle stream of air and a steam bath. Add
5 ml carbon disulfide and again evaporate to dryness to remove
residual benzene. Dissolve residue, transfer to a 25 ml volumetric
flask, and make to volume with carbon disulfide. Add a small amount
of anhydrous sodium sulfate to absorb any water and to clarify the
solution, (final cone 6 mg phenothiazine/ml)
-------
Phenothiazlne EPA-1
(Tentative)
IR Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan both the standard and sample solutions
from 1430 cm" to 1250 cm" (7.0 ^i to 8.0 ji). For a qualitative
comparison, run a full scan.
Determine the absorbance of standard and sample using the
minimum absorbance at 1333 cm (7.5 p) and the maximum absorbance
at 1299 cm"1 (7.7 ju) .
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent phenothiazine as follows:
2 m (a^3- sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
-------
August 1975 Phorate EPA-1
Determination of Phorate
by Infrared Spectroscopy
Phorate is the acceptable common name for 0,0-diethyl S-(ethyl-
thiomethyl) phosphorodithioate, a registered insecticide having the
chemical structure:
CH-^ CHo ^\ 5
3 ^ \ it
CH3—CH2—0'
•P S (-HpS CH-o—CH-7
Molecular formula: C H 0 PS
Molecular weight: 260.4
Melting and boiling point: mp -42.9°C (pure material)
bp 118-120°C at 0.8 mm Hg
Physical state and color: clear mobile liquid
Solubility: 50 ppm in water at RT; miscible with alcohols, esters,
ethers, carbon tetrachloride, dioxane, xylene, and
vegetable oils
Stability: stable at room temperature but is hydrolyzed in the
presence of moisture and by alkalis; incompatible with
alkaline pesticides
Other names: Thimet (American Cyanamid Co.); timet (common name in
USSR), Rampart
-------
2 Phorate EPA-1
Reagents:
1. Phorate standard of known % purity
2. Acetonitrile, pesticide or spectro grade
3. Carbon disulfide, pesticide or spectro grade
4. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Water bath
*
5. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.1 gram phorate standard into a 10 ml volumetric
flask, make to volume with chloroform, and mix well. Add a
small amount of anhydrous sodium sulfate to insure dryness.
(final cone 10 mg/ml)
-------
3 Phorate EPA-1
Preparation of Sample:
Weigh a portion of sample equivalent to 0.5 gram phorate
into a glass-stoppered flask or screw-cap bottle. Add 50 ml
acetonitrile by pipette and 1-2 grams anhydrous sodium sulfate.
Close tightly and shake one hour. Allow to settle; centrifuge
or filter if necessary, taking precaution to prevent evapora-
tion. Evaporate a 10 ml aliquot on a water bath at 40°C with
a stream of dry air blowing across the surface; remove immed-
iately after the last trace of acetonitrile has evaporated.
Dissolve in a small amount of carbon disulfide, transfer to a
10 ml volumetric flask, make to volume, and mix well. Add a
small amount of anhydrous sodium sulfate to insure dryness.
(final cone 10 mg phorate/ml)
Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative settings for the particular IR instrument
being used, scan both the standard and sample from 730 cm to
592 cnT1 (13.7 ji to 16.9 ju) .
Determine the absorbance of standard and sample using the
peak at 654 cm (15.3 u) and baseline from 709 cm to 599 cm
(14.1 ji to 16.7 ju).
Calculation:
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent of phorate as
follows:
7 - Cabs* sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg phorate/ml carbon disulfide gives
an absorbance of approx. 0.03 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
-------
February 1976 Phosphorus Compounds EPA-1
Determination of Total Phosphorus in Pastes
and Organophosphate Formulations
(Acid Digestion and Gravimetric Procedure)
Inorganic phosphorus has been used as an insecticide and rodenticide
in pastes made by grinding yellow phosphorus in the presence of water and
mixing with flour; sometimes glycerin is added.
Organophosphorus compounds of several types have been and are used
as pesticides. These compounds are anticholinesterase chemicals and may
involve danger for the applicator. Examples of the leading series are
as follows (where R represents an organic radical) :
CH3—Ck 0
Phosphate: (dicrotophos) yP O R
CH3—CT
CH3-CH2-0 S
Phosphorothioate : (parathion) /P—0—R
CH3-CH2—CT
CH3-CH2—0 S
Phosphorodithioate: (phorate) /P—S—R
CH3-CH2—0'
There are many analytical methods of different types available for
Organophosphorus compounds; however, there are times when a total phos-
phorus determination is the only immediate means of analysis: e.g., new
compounds, combinations difficult to separate, analytical standard not
available, etc. For data on these compounds, check other methods,
reference book, or company sale literature and data sheets.
-------
2 Phosphorus Compounds EPA-1
Phosphorus exists in three allotropic forms as follows:
(1) White phosphorus (also called yellow phosphorus):
Molecular (atomic) formula: P
Molecular (atomic) weight: 30.975
Melting/boiling point: mp 44.1°C, bp 280°C (volatile, sublimes in
vacuo at ordinary temperature when exposed
to light)
Physical state and color: white or yellow, soft waxy solid or
transparent crystals
Solubility: insoluble in water and alcohol; moderately soluble in
chloroform and benzene; very soluble in carbon disulfide
Stability: at RT it exhibits phosphorescence (slow, luminous
oxidation) in air; it ignites spontaneously in moist
air at about 30°C; stored and shipped beneath water
to avoid ignition. It is very poisonous and causes
severe burns.
(2) Red phosphorus: violet-red, amorphous powder, obtained from white
phosphorus by heating in the presence of a catalyst:
nonpoisonous and much less reactive than the white form;
ignites in air at about 260°C; insoluble in organic
solvents.
(3) Black phosphorus: black, lustrous crystals resembling graphite:
obtained by heating white phosphorus under high pressure;
does not catch fire spontaneously; insoluble in organic
solvents.
Principle of Me_thp_d_:_
Organic matter is destroyed and the phosphorus is oxidized to phos-
phoric acid by a wet acid digestion. The phosphorus is precipitated as
-------
3 Phosphorus Compounds EPA-1
ammonium phosphomolybdate, filtered, washed free from impurities, redis-
solved, and then precipitated as magnesium ammonium pyrophosphate, which
is filtered, washed, and ignited to magnesium pyrophosphate. From the
amount of magnesium pyrophosphate present, the percent phosphorus or
organophosphorus compound may be calculated.
Reagents:
1. Fuming nitric acid, ACS
2. Concentrated sulfuric acid, ACS
3. Concentrated nitric acid, ACS
4. Ammonium nitrate solution - dissolve 100 grams of phosphate-
free ammonium nitrate, ACS in water and make to 1 liter.
5. Concentrated ammonium hydroxide, ACS
6. Ammonium molybdate solution - dissolve 100 grams molybdic acid,
ACS in dilute ammonium hydroxide (144 ml cone, ammonium hydroxide
+ 271 ml water); pour this solution slowly and with constant
stirring into dilute nitric acid (489 ml concentrated nitric
+ 1148 ml water). Keep the mixture in a warm place for several
days or until a portion heated to 65°C deposits no yellow pre-
cipitate of ammonium phosphomolybdate. An alternative procedure
is to heat to 65°C for 1-2 hours and allox<7 to cool and stand
overnight. Decant the solution from any sediment into a clean
glass bottle with a glass stopper or a teflon-lined cap.
7. Ammonium hydroxide 1+1
8. Hydrochloric acid, dilute
9. Magnesia Mixture - dissolve 55 grams of crystallized magnesium
chloride hexahydrate ACS in water; add 140 grams ammonium
chloride ACS and 130.5 ml ammonium hydroxide, and dilute to 1
liter. This solution may form a precipitate if stored for a
long time.
-------
4 Phosphorus Compounds EPA-1
Equipment:
1. Kjeldahl flasks, 500 ml or 800 ml
2. Meker burner
3. Asbestos board with 1.5"-2.0" hole
4. Digestion rack or ring stand and flask support
5. Fume hood
6. Glass beads, small
7. Dropper
8. Filter paper, Whatman No. 7 (special for ammonium phosphoraolybdate
precipitate)
9. Platinum Gooch crucible
10. Asbestos, acid and alkali washed (preferably pre-ignited at
900°-1000°C before washing)
11. Muffle furnace
12. Usual laboratory glassware
Procedure:
Preparation of Sample - Phosphorus Pastes:
Weigh quickly an amount of well mixed sample equivalent to 0.02
gram phosphorus in a 500-800 ml Kjeldahl flask and immediately add
15 ml of water to prevent oxidation by air.
In phosphorus pastes, the phosphorus has a tendency to settle to
the bottom; therefore, it is very important to thoroughly mix the
entire sample before taking a portion for analysis.
A portion of the sample may conveniently be weighed in a No. 11
gelatin capsule and transferred to the digestion flask.
-------
5 Phosphorus Compounds EPA-1
Preparation of Sample - Organophosphate Formulations:
Transfer a weight of sample or an aliquot from a chloroform
extract equivalent to about 0.02 gram of phosphorus into a 500 ml
or 800 ml Kjeldahl flask.
This method is applicable to aerosols, liquid formulations,
emulsifiable concentrates, and chloroform extracts of granules,
dusts, and wettable powders. For the analysis of organophosphates
in granules, dusts, or wettable powders,it is recommended that the
sample be extracted with chloroform. This will simplifv digestion
and avoid detection of inorganic phosphates when the organophos-
phates only are of interest.
Samples may be extracted on a Soxhlet or shaken out with
solvent as follows: a portion of sample not to exceed 50 grams may
be shaken for 2 hours on a shaking machine with 200 ml of chloroform
in a 300 ml screw-cap bottle. After settling or filtering, an
aliquot of the chloroform solution is taken for analysis.
For large aliquots of chloroform extracts or large samples con-
taining petroJeum hydrocarbons, add 25 ml of water before adding
the sulfuric and nitric acids. Evaporate as much as possible of the
organic solvent on a steam bath before digesting over a flame.
Digestion:
For Phosphorus Pastes - place flask on a digestion rack equipped
with an asbestos board having an opening of 1.5-2 inches diameter.
Add 20 ml fuming nitric acid, a few ml at a time, mixing gently but
thoroughly after each addition of acid. A vigorous reaction will
take place. When this action has subsided, heat on a steam bath
until the dense nitric oxide fumes have been expelled. (Use of
nitric acid alone in the initial stages of digestion is desirable
since sulfuric acid will char hydrocarbons and increase the digestion
time and difficulty.) Add 6 small glass beads and 10ml sulfuric acid,
and continue the digestion as belox^, beginning "Continue the addition
-------
6 Phosphorus Compounds EPA-1
For Organoghosphorus Compounds - add 5 ml concentrated sulfuric
acid and mix by swirling; cautiously add concentrated nitric acid,
a few drops at a time, until any vigorous reaction is complete;
then add 5 ml in excess. Add 6 small glass beads and place flask
on a digestion rack equipped with an asbestos board having an opening
of 1.5"-2.0". Heat gently at first over a free flame until the
dense nitric oxide fumes have been expelled. Add a few drops of
nitric acid and heat more vigorously.
Continue the addition of nitric acid and heating until all organic
matter is destroyed, as evidenced by a colorless or light yellow
solution that no longer turns dark. White fumes of sulfur trioxide
will begin to show, and addition of a drop of nitric acid will cause
a sputtering and dense brown fumes. Boil a few minutes to expel any
nitric oxide fumes. Cool, add 10 ml of water, and heat to SO fumes.
Recool; add another 10 ml of water. If brown fumes appear, again
heat to SO fumes.
Allow to cool, add about 25 ml water, and recool. Transfer quan-
titatively to a 600 ml beaker, filtering if not clear. Add 50 ml of
ammonium nitrate solution (or 5 grams solid ammonium nitrate if
volume of solution is over 150 ml). Dilute to about 200 ml.
Pr ecipit at ion as Ammonium Molybdate:
Add ammonium hydroxide to slight alkalinity and then make dis-
tinctly acid with nitric acid. Heat to 65°C and add 70 ml of ammonium
molybdate solution. Stir and digest at 65°C for 30 minutes or longer
if necessary to obtain a clear supernatant liquid. Determine if the
phosphorus has been completely precipitated by adding more molybdate
reagent to the supernatant liquid as soon as it has cleared.
Filter and wash five times by decantation with the ammonium nitrate
solution. The ammonium phosphomolybdate precipitate may be left in
the beaker and washed by decantation or it may be all transferred to
the filter paper and washed there. Test the filtrate with more ammonium
molybdate solution to make certain that enough has been used to pre-
cipitate all of the phosphorus.
-------
7 Phosphorus Compounds EPA-1
Precipitation as Magnesium Ammonium Phosphate:
Dissolve the precipitate on the filter with ammonium hydroxide
(1 4- 1) into the beaker in which the precipitate was formed. Wash
the filter with hot water and rinse off the outside of the filter
funnel stem. Add sufficient ammonium hydroxide to dissolve all
the precipitate and dilute to about 100 ml with water. Neutralize
with hydrochloric acid. Phenolphthalein may be used as an internal
indicator. If the solution is made too acidic, a yellow precipitate
will begin to form. If this happens, add ammonium hydroxide until
precipitate redissolves. Cool and add 20 ml magnesia mixture very
slowly (one drop per second) with constant stirring. Allow to
stand 15 minutes, add 15 ml concentrated ammonium hydroxide, and
to stand overnight or two hours in an ice bath.
Filtration and Ignition:
Filter through a prepared and tared platinum Gooch crucible,
previously ignited for 30 minutes in a muffle furnace at a tempera-
ture of 900-1000°C. Wash with ammonium hydroxide (1 + 9) until free
from chlorides as shown by testing a portion of the acidified filtrate
with silver nitrate. Dry and ignite for 30 minutes at 900-1000°C
until the residue is white. Cool and weigh as magnesium pyrophosphate,
Calculation:
From the weight of magnesium pyrophosphate, MgP^O , calculate
the percent phosphorus in the sample as follows:
(grams MgP 0 )(0.2783)(100)
% phosphorus = -. ;—«
(grams sample)
0.2783 = factor Mg2P207 to phosphorus
-------
Phosphorus Compounds EPA-1
Calculate the percent organophosphate from the percent phosphorus
as follows:
% organophosphate =
% phosphorus
% phosphorus in the organophosphate
% organophosphate = % P X factor P to compound
Reactions involved in this method:
HNO + H SO
Organophosphate — --
Oxidati°n product
H.PO. + 12(NH.)_MoO. + 21HNO, -- -^ (NH. ) ,PO. .12MoO, + 21NH.NO. + 12H.O
34 424 3 434 3 43 2
P04.12Mo03 + 24NH4OH
H2°
-------
July 1975
Picloram EPA-1
(Tentative)
Determination of Picloram by
High Pressure Liquid Chromatography
Picloram is the common name for 4-amino-3,5,6-trichloropicolinic
acid, a registered herbicide having the chemical structure:
NH-
Cl
Molecular formula: C.H.Cl.N-O-
O J j L 2.
Molecular weight: 241.5
Physical state, color, and odor: white powder, chlorine-like odor
Melting point: decomposes before melting
Solubility: 430 ppm in water at 25°C; low in most organic solvents,
1.98 g/100 ml in acetone, 0.55 g/100 ml isopropanol,
less than 50 ppm in carbon disulfide; potassium salt
40% in water
Stability: decomposes approximately 215°C; subject to photo-
decomposition
Other names: Tordon (Dow Chem. Co.), Borolin
Reagents:
1. Picloram standard of known % purity
2. Methanol, pesticide or spectro grade
-------
2 Picloram EPA-1
(Tentative)
Equipment:
1. High pressure liquid chromatograph
2. High pressure liquid syringe or sample injection loop
3. Liquid chromatographic column,2.1 mm I.D. x 1 meter packed
with an anion exchange material such as DuPont's Permaphase
AXX - a quaternary amine bonded to the support by Si-O-Si
linkages
4. Usual laboratory glassware
Operating conditions for DuPont Model 830 LC;
Mobile phase: 90% water (containing 0.2 gram H-PO^ per liter -
approx. 0.003M) + 10% methanol
Column temperature: 65°C
Column pressure: 900 PSI
Flow rate: 8 ml/min
Chart speed: 10 min/inch
Detector: UV at 254 nm
-2
Attenuation: 4 x 10
Conditions may have to be varied by the analyst for other instru-
ments, column variations, sample composition, etc. to obtain optimum
response and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.1 gram picloram standard into a small glass-
stoppered flask or vial, add 10 ml methanol by pipette,
dissolve,and mix well (final cone 10 mg/ml).
-------
3 Picloram EPA-1
(Tentative)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.1 gram picloram
into a small glass-stoppered flask or vial, add 10 ml methanol
by pipette,and shake thoroughly to dissolve the picloram.
Allow any solid matter to settle; filter or centrifuge if
necessary (final cone 10 mg/ml).
Determination:
Alternately inject three 10 ul portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent picloram as follows:
"/ m (pk. ht. or area sample) (wt. std injected) (% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method developed by Elmer H. Hayes, EPA, Beltsville, Md.
-------
November 1975
Pindone EPA-1
Determination of Pindone in Baits and Concentrates
by Ultraviolet Spectroscopy (Ether Extraction)
Pindone is the common name for 2-pivalyl-l,3-indandione, a
registered rodenticide and insecticide having the chemical structure:
Molecular formula: Cn/H,.0.
14 14 3
Molecular weight: 230.3
Melting point: 108.5 to 110.5°C
Physical state and color: yellow crystalline solid
Solubility: 18 ppra in water at 25°C; soluble in most organic solvents;
soluble in aqueous alkali or ammonia to form bright yellow
salts
Stability: stable under normal conditions
Other names: Pivalyl Valone, Pival, Pivalyn (Kilgore Chem. Co.);
pivaldione (France), pival (Portugal, Turkey)
This method may be used for analyzing both bait materials and
concentrates containing about 0.025% and 0.5% active ingredient.
-------
2 Pindone EPA-1
The method does not distinguish between pindone (2-pivalyl-l,3-
indandione) and PMP (2-isovaleryl-l,3-indandione), both of which have
UV absorption maxima at 283, 312, and 324 nm. However, they may be
identified by extraction from the formulation with ether (ethyl or
petroleum), evaporation of the solvent, recrystallization from pentane,
and determination of the melting point. Pindone melts at 110-111°C
and PMP melts at 67-68°C.
Sodium or calcium salts cannot be determined by the ether extrac-
tion method (EPA-1) but can be determined by the pyrophosphate extrac-
tion method (EPA-2).
Reagents:
1. Pindone standard of known % purity
2. Sodium pyrophosphate, 1% solution - dissolve 5 grams
Na.P_0,.10H_0 in 500 ml water.
427 2
3. Ethyl ether, ACS
4. Petroleum ether - extract 200 ml petroleum ether three times
with 20 ml of 1% sodium pyrophosphate solution.
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Soxhlet extraction apparatus
3. Mechanical shaker
4. Centrifuge with 16 x 150 mm glass-stoppered tubes
5. Usual laboratory glassware
-------
3 Pindone EPA-1
Procedure:
Preparation of Standard:
Weigh 0.08 gram pindone standard into a 100 ml volumetric
flask, dissolve in, and make to volume with 1% sodium pyrophos-
phate solution. Mix thoroughly, pipette 10 ml into a 100 ml
volumetric flask, make to volume with pyrophosphate solution,
and mix thoroughly. Pipette 5 ml into a second 100 ml volu-
metric flask, make to volume with pyrophosphate solution, and
mix well. (final cone 4 ug pindone/ml)
Preparation of Sample;
For 0.025% Baits - weigh 16 grams ground sample (0.004 g
pindone) into a Soxhlet thimble, plug with cotton or glass wool,
and extract with ethyl ether on a Soxhlet apparatus for about
four hours. Cool, transfer the extract to a 200 ml volumetric
flask, and make to volume with ethyl ether. Mix thoroughly.
For 0.5% Concentrates - weigh 0.8 gram ground sample (0.004 g
pindone) into a 500 ml glass-stoppered flask, add 200 ml ethyl
ether by pipette, and shake on a mechanical shaker for 30 minutes.
Centrifuge a portion of the extract to clarify if necessary, taking
care to avoid evaporation of ether.
Pipette 2 ml of the clear ether solution into a 16 x 150 mm
glass-stoppered centrifuge tube. Add 10 ml of 1% sodium pyro-
phosphate solution by pipette, shake vigorously for two minutes,
and centrifuge at high speed until the aqueous layer is clear.
Draw off the ether layer and any remaining emulsion using an
aspirator with a glass tube drawn into a fine tip. Add 2 ml ethyl
ether, shake, centrifuge, and draw off the ether. Repeat twice more
with 2 ml portions of petroleum ether. If the aqueous layer is not
clear, centrifuge for a few minutes with the stopper off to remove
any residual ether. (final cone 4 pg pindone/ml)
-------
Pindone EPA-1
UV Determination:
With the UV spectrophotometer at the optimum quantitative
settings for the particular instrument being used, balance the
pen for 0 and 100% at 283 nm with 1% pyrophosphate solution in
each cell. Scan both standard and sample from 350 nm to 200 nm
with the pyrophosphate solution in the reference cell.
Calculation:
Measure the absorbance of standard and sample at 283 nm and
calculate the percent pindone as follows:
7 _ (abs. sample)(cone, std injjg/ml)(% purity std)
(abs. std)(cone, sample in pg/ml)
or using dilution factors, as follows:
2 (abs. sample)(wt. std)(purity std)(1/100)(10/100)(5/100)(100)
(abs. std)(1/200)(2/10)
-------
November 1975
Pindone EPA-2
Determination of Pindone in Baits and Concentrates
by Ultraviolet Spectroscopy (Pyrophosphate Extraction)
Pindone is the common name for 2-pivalyl-l,3-indandione, a
registered rodenticide and insecticide having the chemical structure:
Molecular formula: ^../H ,0
Molecular weight: 230.3
Melting point: 108.5 to 110.5°C
Physical state and color: yellow crystalline solid
Solubility: 18 ppm in water at 25°C; soluble in most organic solvents;
soluble in aqueous alkali or ammonia to form bright yellow
salts
Stability: stable under normal conditions
Other names: Pivalyl Valone, Pival, Pivalyn (Kilgore Chem. Co.),
pivaldione (France), pival (Portugal, Turkey)
This method may be used for analyzing both bait materials and
concentrates containing about 0.025% and 0.5% active ingredient.
-------
2 Pindone EPA-2
The method does not distinguish between pindone (2-pivalyl-l,3-
indandione) and PMP (2-isovaleryl-l,3-indandione), both of which have
UV absorption maxima at 283, 312, and 324 nm. However, they may be
identified by extraction from the formulation with ether (ethyl or
petroleum), evaporation of the solvent, recrystallization from pentane,
and determination of the melting point. Pindone melts at 110-111°C
and PMP melts at 67-68°C.
Sodium or calcium salts cannot be determined by the ether extrac-
tion method (EPA-1) but can be determined by the pyrophosphate extrac-
tion method (EPA-2).
Reagents:
1. Pindone standard of known % purity
2. Sodium pyrophosphate, 1% solution - dissolve 5 grams
Na P 0 .10H 0 in 500 ml water.
3. Ethyl ether-petroleum ether (20-80) - extract 200 ml petroleum
ether three times with 20 ml portions of pyrophosphate solution
and add 50 ml ethyl ether.
4. Hydrochloric acid, 2.5N solution
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording with
matched 1 cm cells
2. Mechanical shaker
3. Centrifuge with 100 ml glass-stoppered centrifuge tubes
4. Usual laboratory glassware
-------
3 Pindone EPA-2
Procedure:
Preparation of Standard;
Weigh 0.08 gram pindone standard into a 100 ml volumetric
flask, dissolve in, and make to volume with 1% sodium pyrophos-
phate solution. Mix thoroughly and pipette 10 ml into a 100 ml
volumetric flask, make to volume with pyrophosphate solution,
and mix thoroughly. Pipette 5 ml into a second 100 ml volumetric
flask, make to volume with pyrophosphate solution, and mix well.
(final cone 4 ug pindone/ml)
Preparation of Sample:
Weigh an amount of finely ground sample equivalent to 0.001
gram pindone (4 grams of 0.025% Bait or 0.2 gram 0.5% Concentrate)
into a 250 ml glass-stoppered flask, add by pipette 100 ml 1%
sodium pyrophosphate solution, and shake on a mechanical shaker
for one hour. Transfer 40-50 ml to a glass-stoppered centrifuge
tube and centrifuge for at least 5 minutes. Pipette 20 ml of
this solution into a glass-stoppered 100 ml centrifuge tube, add
5 ml 2.5N hydrochloric acid and 50 ml (by pipette) of the mixed
ether solution, and shake for five minutes. If an emulsion forms,
centrifuge to break the emulsion. Pipette 10 ml of the ether layer
to a clean centrifuge tube and add 10 ml pyrophosphate solution by
pipette. Shake for 2 minutes and remove the ether layer using an
aspirator with a glass tube drawn to a fine tip. If the aqueous
layer is not clear, centrifuge for a few minutes with the stopper
off to remove any residual ether. (final cone 4 ug pindone/ml)
UV Determination:
With the UV spectrophotometer at the optimum quantitative
settings for the particular instrument being used, balance the pen
for 0 and 100% at 283 nm with 1% pyrophosphate solution in each
cell. Scan both standard and sample from 350 nm to 200 nm with
the pyrophosphate solution in the reference cell.
-------
Pindonc EPA-2
Calculation:
Measure the absorbance of standard and sample at 283 nm and
calculate the percent pindone as follows:
7 a (abSy sample)(cone, std in ug/ml)(% purity std)
(abs. std) (cone, sample in jig/ml)
or using dilution factors, as follows:
(abs. sample)(wt. std)(purity std)(1/100)(10/100)(5/100)(100)
" (abs. std)(wt. sample)(1/100)(20/50)(10/10)
% Sodium pindone « % pindone X 1.096
-------
November 1975
Pindone EPA-3
Determination of Pindone in Water-Soluble Formulations
by Ultraviolet Spectroscopy (Pyrophosphate Extraction)
Pindone is the common name for 2-pivalyl-l,3-indandione, a
registered rodenticide and insecticide having the chemical structure:
Molecular formula: C,.H..O_
14 14 3
Molecular weight: 230.3
Melting point: 108.5 to 110.5°C
Physical state and color: yellow crystalline solid
Solubility: 18 ppm in water at 25°C; soluble in most organic solvents;
soluble in aqueous alkali or ammonia to form bright yellow
salts
Stability: stable under normal conditions
Other names: Pivalyl Valone, Pival, Pivalyn (Kilgore Chem. Co.);
pivaldione (France), pival (Portugal, Turkey)
Pindone (2-pivalyl-l,3-indandione) and PMP (2-isovaleryl-l,3-
indandione) are often formulated as water-soluble powders containing
the sodium salts of these two materials, along with sodium benzoate,
-------
2 Pindone EPA-3
sodium ethylenediamine tetraacetate (EDTA), and sugar. Sodium benzoate
and the sodium EDTA interfere moderately at the strongest absorption
maxima near 283 nm, decreasing to about 275 run and then increasing
again; however, a determination can be made at the secondary maxima
near 311 and 323 nm.
A solution of pindone containing 10 ug/ml in 1% pyrophosphate has
an approximate absorbance of 0.394 at 324 nm; a solution of PMP con-
taining 7.5 jag/ml in 1% pyrophosphate has an approximate absorbance of
0.398 at 323 nm. If there is no interference and the absorbances are
read at 283 nm, the concentrations of the standards and/or sample
solutions should each be about one-third as great.
This method does not distinguish between pindone and PMP; however,
they may be identified by extracting an acidified aqueous solution of
the formulation with ether (ethyl or petroleum), evaporating the solvent,
recrystallizing from pentane, and determining the melting point. Pindone
melts at 110-111°C and PMP melts at 67-68°C.
The presence of sodium benzoate or sodium EDTA may be confirmed by
the procedure at the end of this method.
Reagents:
1. Pindone standard of known % purity
2. Sodium pyrophosphate, 1% solution - dissolve 10 grams
Na,P20 .10H-0 in one liter of water.
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Usual laboratory glassware
-------
3 Pindone EPA-3
Procedure:
Preparation of Standard:
Weigh 0.1 gram pindone standard into a 100 ml volumetric
flask; dissolve and make to volume with 1% sodium pyrophosphate
solution. Mix thoroughly and pipette 5 ml into a 50 ml volu-
metric flask, and make to volume with pyrophosphate solution;
mix well, pipette 5 ml into a second 50 ml volumetric flask,
and make to volume with pyrophosphate solution. (final cone
10 jag pindone/ml)
If absorbances are to be read at 283 nm, pipette 2 ml
instead of 5 ml into the second 50 ml volumetric flask. (final
cone 4 jig pindone/ml)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.004 gram pindone
into a 100 ml volumetric flask; dissolve and make to volume
with 1% sodium pyrophosphate solution. Mix thoroughly, pipette
25 ml into a second 100 ml volumetric flask, and make to volume
with the 1% pyrophosphate solution. (final cone 10 pg pindone/ml)
If the absorbances are to be read at 283 nm, pipette 10 ml
instead of 25 ml into the second 100 ml volumetric flask, (final
cone 4 |j,g pindone/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
settings for the particular instrument being used, balance the
pen for 0 and 100% at 324 nm (or at 283 nm if no sodium benzoate
or sodium EDTA interference is present) with 1% pyrophosphate
solution in each cell. Scan both the standard and sample from
350 nm to 250 nm with pyrophosphate solution in the reference
cell.
-------
4 Pindone EPA-3
Calculation:
Measure the absorbance of standard and sample at 324 nm
or 283 nm, and calculate the percent pindone as follows:
% pindone at 324 nm:
(abs. sample)(wt. std)(purity std)(1/100)(5/50)(5/50)
= (abs. std)(wt. sample)(1/100)(25/100)
% pindone at 283 nm:
(abs. sample)(wt. std)(purity std)(1/100)(5/50)(2/50)
= (abs. std)(wt. sample)(1/100)(10/100)
Procedure for Confirming the Presence of Sodium Benzoate
and Sodium EDTA;
Sodium benzoate and sodium EDTA may be identified by the
following procedure: Make an aqueous extract of the sample,
acidify with hydrochloric acid, and filter. Save both filtrate
and residue. Use the filtrate for EDTA and the residue for
benzoate as follows:
For EDTA - place one drop of nickel sulfate solution (0.01%
in water) and one drop concentrated ammonium hydroxide into each
of two depressions on a spot plate. To one add a drop of water
and to another a drop of the sample extract filtrate. Add a drop
of dimethylglyoxime solution (saturated - approx. 0.1 g in 50 ml
water) to each. The blank becomes pink immediately, but if the
solution contains a sequestering agent—EDTA—it remains color-
less or becomes only very faintly pink.
-------
Pindone EPA-3
For Benzoate - wash the residue with hot water to remove the
benzoic acid (pindone and PMP are practically insoluble in water).
Make alkaline with a few drops of ammonium hydroxide, heat gently
to expel excess ammonia by evaporation, dissolve residue in a
few ml water (filter if necessary), and add a few drops of aqueous
0.5% ferric chloride solution. A salmon-color precipitate of
ferric benzoate indicates presence of benzoic acid. An alterna-
tive procedure is to evaporate the acidified filtrate and
determine the melting point. Benzoic acid melts at 122°C.
-------
January 1976
Piperonyl Butoxide EPA-1
Detection of Piperonyl Butoxide
in Pesticides - Qualitative Test
Piperonyl butoxide, technical is the official name for the commercial
product consisting of 80% (butyl carbityl)(6-propylpiperonyl) ether and
20% related compounds.
Piperonyl butoxide is a registered pesticide ingredient and, although
itself without marked insecticidal properties, enhances the toxicity,
paralytic effect, and persistent contact toxicity of the pyrethrins and
related compounds. It is also used with rotenone and tetramethrin.
The chemical structure is:
•CH2-CH2-CH3
Molecular formula: C -H.-O.
Molecular weight: 338.5
Boiling point: 180°C at 1 mm Hg
Physical state, color, and odor: odorless, pale yellow oily liquid
Solubility: soluble in most organic solvents including petroleum oils
and dichlorodifluoromethane; insoluble in water
Stability: stable to light; resistant to hydrolysis; non-corrosive
-------
2 Piperonyl Butoxide EPA-1
Other names: Butacide (FMC), NIA 5273 (Niagara), FMC 5273, Q"-[2-(2-n-
butoxyethoxy)-ethoxy]-4,5-methylenedioxy-2-propyltoluene
Reagents:
1. Tannir acid
2. Acetic acid, glacial
3. Phosphoric acid, 85%
4. Color development reagent - dissolve completely 0.05 gram tannic
acid in 15 ml glacial acetic acid, add 35 ml 85% phosphoric acid,
and mix thoroughly. Prepare fresh daily and keep in tightly
stoppered bottle since the solution is hygroscopic.
Stock solutions of tannic acid in acetic acid (solution A)
and phosphoric acid (solution B) may be kept separately and mixed
1.5mlA+3.5mlB just before use.
Equipment:
1. 18 x 150 mm test tube
2. Boiling water bath
3. Usual laboratory glassware
Preparation of Sample;
The sample to be tested should contain 1-2 mg per ml of solution.
Oil solutions may be diluted with ether or an odorless base oil
such as Deo Base.
Powders should be extracted with ethyl ether by shaking in a flask
on a shaking machine and evaporated or diluted to the desired concentration.
Qualitative Determination:
Place 0.1 ml of sample solution and 5 ml of color reagent in an
18 x 150 mm test tube. Shake the tube vigorously for 30 seconds and place
in a bath of boiling water for 5 minutes. A blue color indicates the
presence of piperonyl butoxide.
-------
January 1976
Piperonyl Butoxide EPA-2
Determination of Piperonyl Butoxide
by Gas-Liquid Chromatography
(FID - Internal Standard)
Piperonyl butoxide, technical is the official name for the commercial
product consisting of 80% (butyl carbityl)(6-propylpiperonyl) ether and
20% related compounds.
Piperonyl butoxide is a registered pesticide ingredient and, although
itself without marked insecticidal properties, enhances the toxicity,
paralytic effect, and persistent contact toxicity of the pyrethrins and
related compounds. It is also used with rotenone and tetramethrin.
The chemical structure is:
P—
'o—
•CH2-CH2-CH3
CH2-0-CH2-CH2-0- CH2-CH2-0- C H9
Molecular formula: C H 0
Molecular weight: 338.5
Boiling point: 180°C at 1 mm Hg
Physical state, color, and odor: odorless, pale yellow oily liquid
Solubility: soluble in most organic solvents including petroleum oils
and dichlorodifluoromethane; insoluble in water
Stability: stable to light; resistant to hydrolysis; non-corrosive
-------
2 Piperonyl Butoxide EPA-2
Other names: Butacide (FMC), NIA 5273 (Niagara), FMC 5273, or-[2-(2-n-
butoxyethoxy)-ethoxy]-4,5-methylenedioxy-2-propyltoluene
Reagents:
1. Piperonyl butoxide of known % purity
2. Dioctyl phthalate
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - veigh 0.18 gram dioctyl phthalate
into a 100 ml volumetric flask; dissolve in and make to volume
with acetone. (cone 1.8 mg dioctyl phthalate/ml)
Equipment:
1. Gas chroraatograph with flame ionization detector (FID)
2. Column: 6' x 4 mm glass column packed with 3% OV-1 on 60/80 mesh
Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 230°C
Injection temperature: 260°C
Detector temperature: 270°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi (adjusted for specific GC)
Hydrogen pressure: 20 psi (30 ml/min)
Air pressure: 30 psi (300 ml/min)
-------
3 Piperonyl Butoxide EPA-2
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
Procedure:
Preparat ion of Standard;
Weigh 0.03 gram piperonyl butoxide standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 25 ml of the
internal standard solution and shake to dissolve. (final cone 1.2 mg
piperonyl butoxide and 1.8 mg dioctyl phthalate/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.03 gram piperonyl
butoxide into a small glass-stoppered flask or screw-cap bottle.
Add by pipette 25 ml of the internal standard solution. Close
tightly and shake thoroughly to dissolve and extract the piperonyl
butoxide. For coarse or granular materials, shake mechanically for
30 minutes or shake by hand intermittently for one hour. (final
cone 1.2 mg piperonyl butoxide and 1.8 mg dioctyl phthalate/ml)
Determination:
Inject 2 ul of standard and, if necessary, adjust the instrument
parameters and the volume injected to give a complete separation
within a reasonable time and peak heights of from 1/2 to 3/4 full
scale. The elution order is piperonyl butoxide, then dioctyl
phthalate.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of piperonyl butoxide and
dioctyl phthalate from both the standard-internal standard solution
and the sample-internal standard solution.
-------
RF
Piperonyl Butoxide EPA-2
Determine the RF value for each injection of the standard-internal
standard solution as follows and calculate the average:
I.S. = Internal Standard = dioctyl nhthalate
(wt. I.S.)(% purity I.S.)(pk. ht. or area piperonyl butoxide)
(wt. piperonyl butoxide)(% purity piperonyl butoxide)(pk. ht. or area I.S.)
Determine the percent piperonyl butoxide for each injection of
the sample-internal standard solution as follows and calculate the
average:
7 - Cwt. I.S.)(% purity I.S.)(pk. ht. or area piperonyl butoxide)fM3>
(wt. sample) (pk. ht. or area I.S.)(RF) (Ll-l]
Method submitted by Division of Regulatory Services, Kentucky Agricultural
Experiment Station, University of Kentucky, Lexington, Kentucky 40506.
-------
November 1975
PMP EPA-1
Determination of PMP in Baits and Concentrates
by Ultraviolet Spectroscopy (Ether Extraction)
PMP is 2-isovaleryl-l,3-indandione, a registered rodenticide having
the chemical structure:
-CH3
CH2CH
Molecular formula: C..H-.0
Molecular weight: 230.3
Melting point: 67 to 68°C
Physical state and color: yellow crystalline solid
Solubility: practically insoluble in water; soluble in most organic
solvents; soluble in aqueous alkali or ammonia to form
bright yellow salts
Stability: stable under normal conditions
Other names: Valone (Kilgore Chem. Co.)
This method may be used for analyzing both bait materials and
concentrates containing about 0.025% and 0.5% active ingredient.
-------
2 PMP EPA-1
The method does not distinguish between pindone (2-pivalyl-l,3-
indandione) and PMP (2-isovaleryl-l,3-indandione), both of which have
UV absorption maxima at 283, 312, and 324 nm. However, they may be
identified by extraction from the formulation with ether (ethyl or
petroleum), evaporation of the solvent, recrystallization from pentane,
and determination of the melting point. Pindone melts at 110-111°C
and PKP melts at 67-68°C.
Sodium or calcium salts cannot be determined by the ether extrac-
tion method (EPA-1) but can be determined by the pyrophosphate extrac-
tion method (EPA-2).
Reagents:
1. PMP standard of known % purity
2. Sodium pyrophosphate, 1% solution - dissolve 5 grams
Na.P.O-.lOH.O in 500 ml water.
427 2
3. Ethyl ether, ACS
4. Petroleum ether - extract 200 ml petroleum ether three times
with 20 ml of 1% sodium pyrophosphate solution.
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Soxhlet extraction apparatus
3^ Mechanical shaker
4. Centrifuge with 16 x 150 mm glass-stoppered tubes
5. Usual laboratory glassware
-------
3 PMP EPA-1
Procedure:
Preparation of Standard:
Weigh 0.08 gram PMP standard into a 100 ml volumetric flask,
dissolve in, and make to volume with 1% sodium pyrophosphate
solution. Mix thoroughly, pipette 10 ml into a 100 ml volu-
metric flask, make to volume with pyrophosphate solution, and again
mix thoroughly. Pipette 5 ml into a second 100 ml volumetric
flask, make to volume with pyrophosphate solution, and mix well.
(final cone 4 jig PMP/ml)
Preparation of Sample:
For 0.025% Baits - weigh 16 grams ground sample (0.004 g PMP)
into a Soxhlet thimble, plug with cotton or glass wool, and
extract with ethyl ether on a Soxhlet apparatus for about four
hours. Cool, transfer the extract to a 200 ml volumetric flask,
and make to volume with ethyl ether. Mix thoroughly.
For 0.5% Concentrates - weigh 0.8 gram ground sample (0.004 g
PMP) into a 500 ml glass-stoppered flask, add 200 ml ethyl ether
by pipette, and shake on a mechanical shaker for 30 minutes.
Centrifuge a portion of the extract to clarify if necessary,
taking care to avoid evaporation of ether.
Pipette 2 ml of the clear ether solution into a 16 x 150 mm
glass-stoppered centrifuge tube. Add 10 ml of 1% sodium pyro-
phosphate solution by pipette, shake vigorously for two minutes,
and centrifuge at high speed until the aqueous layer is clear.
Draw off the ether layer and any remaining emulsion using an
aspirator with a glass tube drawn into a fine tip. Add 2 ml ethyl
ether, shake, centrifuge, and draw off the ether. Repeat twice
more with 2 ml portions of petroleum ether. If the aqueous layer
is not clear, centrifuge for a few minutes with the stopper off to
remove any residual ether, (final cone 4 ug PMP/ml)
-------
PMP EPA-1
UV Determination:
With the UV spectrophotometer at the optimum quantitative
settings for the particular instrument being used, balance the
pen for 0 and 100X at 283 nm with 1% pyrophosphate solution in
each cell. Scan both standard and sample from 350 nm to 200 nm
with the pyrophosphate solution in the reference cell.
Calculation;
Measure the absorbance of standard and sample at 283 nm and
calculate the percent PMP as follows:
(abs. sample) (cone, std in pg/ml)JJLpJurji^._std_I
" * (abs. std)(cone, sample in pg/ml)
or using dilution factors, as follows:
2 (abs. sample)(wt. std)(purity std)(1/100)(10/100)(5/100)(100)
(abs. std)(1/200)(2/10)
-------
November 1975
PMP EPA-2
Determination of PMP in Baits and Concentrates
by Ultraviolet Spectroscopy (Pyrophosphate Extraction)
PMP is 2-isovaleryl-l,3-indandione, a registered rodenticide
having the chemical structure:
Molecular formula: C .H ,0,
Molecular weight: 230.3
Melting point: 67 to 68*C
Physical state and color: yellow crystalline solid
Solubility: practically insoluble in water; soluble in most organic
solvents; soluble in aqueous alkali or ammonia to form
bright yellow salts
Stability: stable under normal conditions
Other names: Valone (Kilgore Chem. Co.)
This method may be used for analyzing both bait materials and
concentrates containing about 0.025% and 0.5% active ingredient.
-------
2 PMP EPA-2
The method does not distinguish between pindone (2-pivalyl-l,3-
indandione) and PMP (2-isovaleryl-l,3-indandione), both of which have
UV absorption maxima at 283, 312, and 324 nm. However, they may be
identified by extraction from the formulation with ether (ethyl or
petroleum), evaporation of the solvent, recrystallization from pentane,
and determination of the melting point. Pindone melts at 110-111°C
and PMP melts at 67-68°C.
Sodium or calcium salts cannot be determined by the ether extrac-
tion method (EPA-1) but can be determined by the pyrophosphate extrac-
tion method (EPA-2).
Reagents:
1. PMP standard of known % purity
2. Sodium pyrophosphate, 1% solution - dissolve 5 grams
Na.P 0-.10H 0 in 500 ml water.
3. Ethyl ether-petroleum ether (20-80) - extract 200 ml petroleum
ether three times with 20 ml portions of pyrophosphate solution
and add 50 ml ethyl ether.
4. Hydrochloric acid, 2.5N solution
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Centrifuge with 100 ml glass-stoppered centrifuge tubes
4. Usual laboratory glassware
-------
3 PMP EPA-2
Procedure:
Preparation of Standard:
Weigh 0.08 gram PMP standard into a 100 ml volumetric flask,
dissolve in, and make to volume with 1% sodium pyrophosphate
solution. Mix thoroughly and pipette 10 ml into a 100 ml volu-
metric flask, make to volume with pyrophosphate solution, and again
mix thoroughly. Pipette 5 ml into a second 100 ml volumetric
flask, make to volume with pyrophosphate solution, and mix well.
(final cone 4 iig PMP/ml)
Preparation of Sample;
Weigh an amount of finely ground sample equivalent to 0.001
gram PMP (4 grams of 0.025% Bait or 0.2 gram 0.5% Concentrate)
into a 250 ml glass-stoppered flask, add by pipette 100 ml 1%
sodium pyrophosphate solution, and shake on a mechanical shaker
for one hour. Transfer 40-50 ml to a glass-stoppered centrifuge
tube and centrifuge for at least 5 minutes. Pipette 20 ml of this
solution into a glass-stoppered 100 ml centrifuge tube, add 5 ml
2.5N hydrochloric acid and 50 ml (by pipette) of the mixed ether
solution, and shake for five minutes. If an emulsion forms,
centrifuge to break the emulsion. Pipette 10 ml of the ether layer
to a clean centrifuge tube and add 10 ml pyrophosphate solution by
pipette. Shake for 2 minutes and remove the ether layer using an
aspirator with a glass tube drawn to a fine tip. If the aqueous
layer is not clear, centrifuge for a few minutes with the stopper
off to remove any residual ether, (final cone 4 ug PMP/ml)
UV Determination:
With the UV spectrophotometer at the optimum quantitative
settings for the particular instrument being used, balance the pen
for 0 and 100% at 283 nm with 1% pyrophosphate solution in each
cell. Scan both standard and sample from 350 nm to 200 nm with
the pyrophosphate solution in the reference cell.
-------
PMP EPA-2
Calculation:
Measure the absorbance of standard and sample at 283 nm and
calculate the percent PMP as follows:
(abs. sample)(cone. std in ug/ml)(% purity std)
(abs. std)(cone, sample in ug/ml)
% Calcium PMP - % PMP X 1.083
% Sodium PMP « % PMP X 1.096
(anhydrous)
or using dilution factors, as follows:
y = (abs. sample)(wt. 8td)(purity std)(1/100)(10/100)(5/100)(100)
(abs. std)(wt. sample)(1/100)(20/50)(10/10)
-------
November 1975
PMP EPA-3
Determination of PMP in Water-Soluble Formulations
by Ultraviolet Spectroscopy (Pyrophosphate Extraction)
PMP is 2-isovaleryl-l,3-indandione, a registered rodenticide
having the chemical structure:
0 /CHj
II //
C—CH2CH
Molecular formula: C,,H,.0.
14 14 3
Molecular weight: 230.3
Melting point: 67 to 68°C
Physical state and color: yellow crystalline solid
Solubility: practically insoluble in water; soluble in most organic
solvents; soluble in aqueous alkali or ammonia to form
bright yellow salts
Stability: stable under normal conditions
Other names: Valone (Kilgore Chem. Co.)
Pindone (2-pivalyl-l,3-indandione) and PMP (2-isovaleryl-l,3-
indandione) are often formulated as water-soluble powders containing
the sodium salts of these two materials, along with sodium benzoate,
sodium ethylenediamine tetraacetate (EDTA), and sugar. Sodium benzoate
-------
2 PMP EPA-3
and the sodium EDTA interfere moderately at the strongest absorption
maxima near 283 nm, decreasing to about 275 nm and then increasing
again; however, a determination can be made at the secondary maxima
near 311 and 323 nm.
A solution of pindone! containing 10 ug/ml in 1% pyrophosphate
has an approximate absorbance of 0.394 at 324 nm; a solution of PMP
containing 7.5 jig/ml in 1% pyrophosphate has an approximate absorbance
of 0.398 at 323 nm. If there is no interference and the absorbances
are read at 283 nm, the concentrations of the standards and/or sample
solutions should each be about one-third as great.
This method does not distinguish between pindone and PMP; however,
they may be identified by extracting an acidified aqueous solution of
the formulation with ether (ethyl or petroleum), evaporating the solvent,
recrystallizing from pentane, and determining the melting point. Pindone
melts at 110-111'C and PMP melts at 67-68°C.
The presence of sodium benzoate or sodium EDTA may be confirmed by
the procedure at the end of this method.
Reagents:
1. PMP standard of known % purity
2. Sodium pyrophosphate, 1% solution - dissolve 10 grams
Na.P-O-.lOH-O in one liter of water.
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Usual laboratory glassware
-------
3 PMP EPA-3
Procedure;
Preparation j)f_ Standard:
Weigh 0.075 gram PMP standard into a 100 ml volumetric flask;
dissolve and make to volume with 1% sodium pyrophosphate solution.
Mix thoroughly and pipette 5 ml into a 50 ml volumetric flask,
and make to volume with pyrophosphate solution; mix well, pipette
5 ml into a second 50 ml volumetric flask, and make to volume
with pyrophosphate solution. (final cone 7.5 ug PMP/ml)
If absorbances are to be read at 283 nm, pipette 2 ml instead
of 5 ml into the second 50 ml volumetric flask, (final cone 3 ^ig
PMP/ml)
Preparation ofSample:
Weigh an amount of sample equivalent to 0.003 gram PMP into
a 100 ml volumetric flask; dissolve and make to volume with 1%
sodium pyrophosphate solution. Mix thoroughly, pipette 25 ml
into a second 100 ml volumetric flask, and make to volume with
the 1% pyrophosphate solution. (final cone 7.5 ^ig PMP/ml)
If the absorbances are to be read at 283 nm, pipette 10 ml
instead of 25 ml into the second 100 ml volumetric flask, (final
cone 3 jig PMP/ml)
UV Determination:
With the UV spectrophotometer at the optimum quantitative
settings for the particular instrument being used, balance the
pen for 0 and 100% at 324 nm (or at 283 nm if no sodium benzoate
or sodium EDTA interference is present) with 1% pyrophosphate
solution in each cell. Scan both the standard and sample from
350 nm to 250 nm with pyrophosphate solution in the reference
cell.
-------
4 PMP EPA-3
Calculation:
Measure the absorbance of standard and sample at 324 nm
or 283 nm, and calculate the percent PMP as follows:
% PMP at 324 nm:
y (abs. sample)(vt. std)(purity std)(1/100)(5/50)(5/50)
° (abs. std)(wt. sample)(1/100)(25/100)
% PMP at 283 nm:
(abs. sample)(wt. std)(purity std)(1/100)(5/50)(2/50)
(abs. std)(wt. sample)(1/100)(10/100)
Procedure for Confirming the Presence of Sodium Benzoate
and Sodium EDTA:
Sodium benzoate and sodium EDTA may be identified by the
following procedure: Make an aqueous extract of the sample,
acidify with hydrochloric acid, and filter. Save both filtrate
and residue. Use the filtrate for EDTA and the residue for
benzoate as follows:
For EDTA - place one drop of nickel sulfate solution (0.01%
in water) and one drop concentrated ammonium hydroxide into each
of two depressions on a spot plate. To one add a drop of water
and to another a drop of the sample extract filtrate. Add a drop
of dimethylglyoxime solution (saturated - approx. 0.1 g in 50 ml
water) to each. The blank becomes pink immediately, but if the
solution contains a sequestering agent—EDTA—it remains colorless
or becomes only very faintly pink.
-------
PMP EPA-3
For Benzoate - wash the residue with hot water to remove the
benzoic acid (pindone and PMP are practically insoluble in water).
Make alkaline with a few drops of ammonium hydroxide, heat gently
to expel excess ammonia by evaporation, dissolve residue in a few
ml water (filter if necessary), and add a few drops of aqueous
0.5% ferric chloride solution. A salmon—color precipitate of
ferric benzoate indicates presence of benzoic acid. An alterna-
tive procedure is to evaporate the acidified filtrate and
determine the melting point. Benzoic acid melts at 122°C.
-------
November 1975
Prometone EPA-1
(Tentative)
Determination of Prometone
by Gas-Liquid Chromatography
(TCD - Internal Standard)
Prometone is the accepted common name for 2,4-bis (isopropylamino)-
6-methoxy-s-triazine, a registered herbicide having the chemical struc-
ture:
H-,C
Molecular formula: C10H19N5°
Molecular weight:
Melting point:
225.3
91 to 92°C; the technical product is at least 97%
pure and has a ra.p. of 88-90°C
Physical state and color: white crystalline solid
Solubility: 750 ppm in water at 20°C; readily soluble in acetone,
benzene, chloroform, methanol
Stability: stable under neutral or slightly acidic or alkaline
conditions but is hydrolyzed by stronger acid or alkali;
compatible with most other pesticides when used at normal
rates; non-corrosive under normal use conditions
Other names: Primatol, Pramitol, G41435 (Ciba-Geigy); prometon (ISO),
Gesafram, Outrack
-------
2 Prometone EPA-1
(Tentative)
Reagents:
1. Prometone standard of known % purity
2. Technical heptachlor
3. Chloroform, pesticide or spectro grade
4. Internal Standard solution - weigh 0.25 gram technical
heptachlor into a 25 ml volumetric flask; dissolve in and
make to volume with chloroform. (cone 10 mg heptachlor/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 6' x 1/4" OD glass column packed with 4% SE-30 on
80/100 mesh Diatoport S (or equivalent column -
glass should be used because heptachlor degrades
on metal column)
3. Precision liquid syringe: 5 or 10 jil
4. Usual laboratory glassware
Operating Conditions for TCP:
Column temperature: 175°C
Injection temperature: 225°C
Detector temperature: 250°C
Filament current: 225 ma
Carrier gas: Helium
Carrier gas flow rate: 30 ml/min
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and
reproducibility.
-------
Prometone EPA-1
(Tentative)
Procedure:
Preparat ion o^f ™
Weigh 0.025 gram prometone standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 10 ml of
the internal standard solution and shake to dissolve. (final
cone 2.5 mg prometone and ]0 mg heptachlor/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.025 gram prometone
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 10 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the prometone. For
coarse or granular materials, shake mechanically for 30 minutes
or shake by hand intermittently for one hour. (final cone 2.5 mg
prometone and 10 mg heptachlor/ml)
Determinationt
Inject 2-4 pi of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is prometone, then heptachlor,
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Caleula t ion:
Measure the peak heights or areas of prometone and heptachlor
from both the standard--Jntemal standard solution and the sample-
internal standard solution.
-------
Prometone EPA-1
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. heptachlor)(% purityheptachlor)(pk. ht. or area prometone)
(wt. prometone)(% purity prometone)(pk. ht. or area heptachlor)
Determine the percent prometone for each injection of the
sample-internal standard solution as follows and calculate the
average:
„ = (wt. heptachlor)(% purity heptachlor)(pk. ht. or area prometone)
(wt. sample) (pk. ht. or area heptachlor) (RF) '(1.4-1)
This method was developed by Stelios Gerazounis, EPA, Region II, New
York, N. Y., and was collaborated (with slight modification) by Elmer
Hayes, EPA, Beltsville Chemistry Laboratory, Beltsville, Md.
-------
January 1976
Prometone EPA-2
(Tentative)
Determination of Prometone by
Gas—Liquid Chromatography
(FID - Internal Standard)
Prometone is the accepted common name for 2,4-bis (isopropylamino)-
6-methoxy-s-triazine, a registered herbicide having the chemical
structure:
0-CH3
Molecular formula: C ~H gN 0
Molecular weight: 225.3
Melting point: 91 to 92"C; the technical product is at least 97%
pure and has a m.p. of 88-90°C
Physical state and color: white crystalline solid
Solubility: 750 ppm in water at 20°C; readily soluble in acetone,
benzene, chloroform, methanol
Stability: stable under neutral or slightly acidic or alkaline
conditions but is hydrolyzed by stronger acid or alkali;
compatible with most other pesticides when used at normal
rates; non-corrosive under normal use conditions
Other names: Primatol, Pramitol, G 41435 (Ciba-Geigy); prometon (ISO)
Gesafram, Outrack
-------
2 Prometone EPA-2
(Tentative)
Reagents:
1. Prometone standard of known % purity
2. Alachlor standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.5 gram alachlor into a
100 ml volumetric flask; dissolve in and make to volume
with acetone, (cone 5 mg alachlor/ml)
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm glass column packed with 3% OV-17 on
80/100 Gas Chrom Q (or equivalent column)
3, Precision liquid syringe: 5 or 10 jil
4. Mechanical shaker
5. Centrifuge or filtration equipment
6, Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 180°C
Injection temperature: 230°C
Detector temperature: 230eC
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi (adjusted for specific GC)
Hydrogen pressure: 20 psi (adjusted for specific GC)
Air pressure: 30 psi (adjusted for specific GC)
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
3 Prometone EPA-2
(Tentative)
Procedure; (see note after calculations)
Preparation of Standard,:
Weigh 0.05 gram prometone standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 25 ml of the internal
standard solution and shake to dissolve, (final cone 2 mg prometone
and 5 mg alachlor/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.05 gram prometone
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 25 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the prometone. For
coarse or granular materials, shake mechanically for 30 minutes or
shake by hand intermittcmtly for one hour. (final cone 2 mg
prometone and 5 mg alachlor/ml)
Determination:
Inject 1-2 pi of standard and, if necessary, adjust the instru-
ment parameters and the volume injected to give a complete separation
within a reasonable time and peak heights of from 1/2 to 3/4 full
scale. The elution order is prometone, then alachlor.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of prometone and alachlor from
both the standard-internal standard solution and the sample-internal
standard solution.
Determine the RF value for each injection of the standard-internal
standard solution as follows and calculate the average:
-------
4 Prometone EPA-2
(Tentative)
= (wt. alachlor)(% purity alachlor)(pk. ht. or area prometone)
(wt. prometone)(% purity prometone)(pk. ht. or area alachlor)
Determine the percent for each injection of the sample-internal
standard solution as follows and calculate the average:
„ _ (wt. alachlor)(% purity alachlor)(pk. ht. or area prometone)43^ffi>
(wt. sample) (pk. ht. or area alachlor) (RF) U~ l
Note: For an alternative procedure to the above method, the following
changes can be made:
solvent: chloroform
sample concentration: 1.6 rag/ml
column: 4' x 2 mm ID glass, packed with 5% OV-210 on
80/100 Chromosorb W HP
column temperature: 160°
other parameters: adjusted as needed to give optimum
results with the changed conditions
This method was submitted by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond, Virginia
23219.
This method has been designated as tentative since it is a Va. Exp. method
and because some of the data has been suggested by EPA's Beltsville Chem-
istry Lab. Any comments, criticisms, suggestions, data, etc. concerning
this method will be appreciated.
-------
November 1975
Propargite EPA-1
(Tentative)
Determination of Propargite
by Infrared Spectroscopy
Propargite is a common name for 2-(p-tert-butylphenoxy)cyclohexyl-
2-propynyl sulfite, a registered acaricide having the chemical structure;
CH2-CH2
0—CH CH?
\ /
CH2-CH2
0
H C
Molecular formula: C1rtH0,0,S
19 26 4
•CH-
0 — o — O
Molecular weight: 350
Melting or boiling point: (not available)
25
Physical state and color: light to dark amber viscous liquid of d
1.085-1.115; the technical product is at
least 80%
Solubility: practically insoluble in water; soluble in most organic
solvents
Stability: (not available)
Other names: Omite, D014 (Uniroyal); Comite
Reagents:
1. Propargite standard of known % purity
2. Carbon disulfide, pesticide or spectre grade
3. Sodium sulfate, anhydrous, granular
-------
2 Propargite EPA-1
(Tentative)
Equipment :
1. Infrared spectrophotometer, double beam ratio recording, with
matched .5 mm NaCl or KBr cells
2 . Mechanical shaker
3. Rotary evaporator or steam bath
4. Filtration apparatus or centrifuge
5. Usual laboratory glassware
Procedure ;
Preparation of S tandard :
Weigh 0.1 gram propargite standard into a 10 ml volumetric
flask; dissolve in and make to volume with carbon disulfide.
Add a small amount of granular anhydrous sodium sulfate to insure
dryness. (cone 10 mg propargite/ml)
Preparation of Sample;
For dust, granules, and wettable powder , weigh a portion of
sample equivalent to 1 gram propargite into a 250 ml glass-
stoppered Erlenmeyer flask, add by pipette 100 ml carbon disulfide,
stopper, and shake on a mechanical shaker for 1 hour. Allow to
settle; filter or centrifuge if necessary. Add a small amount of
granular anhydrous sodium sulfate to insure dryness. (cone 10 mg
propargite/ml)
liquid formulations and emulsif iable concentrates , weigh
a portion of sample equivalent to 1 gram propargite into a 100 ml
volumetric flask, make to volume with carbon disulfide, and mix
thoroughly. (Interference from solvents in the sample can some-
times be removed by evaporation on a rotary evaporator under
vacuum at about 60°C before making to volume.) Add a small
amount of granular anhydrous sodium sulfate to insure dryness and
clarify the solution. (cone 10 mg propargite/ml)
-------
3 Propargite EPA-1
(Tentative)
An alternative extraction procedure for liquid formulations
and E.C.'s is to shake a 1 gram sample with 100 ml carbon disulfide
and 25-50 ml water in a sealed bottle or flask for 2 hours on a
shaker. Allow to stand for 15 minutes or longer to permit the
carbon disulfide and water layers to separate. With a syringe,
draw off 20-25 ml of carbon disulfide from the bottom of the bottle
and transfer to small vial. Add anhydrous sodium sulfate to insure
dryness and clarify the solution. (cone 10 mg propargite/ml)
IR Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan both the standard and sample from
4000 cm'1 to 3125 cm"1 (2.5 jj to 3.2 ji) .
Determine the a'bsorbance of standard and sample using the peak
at 3300 cm"1 (3.03 p) and a baseline from 3356 cm"1 to 3247 cm
(2.98 p to 3.08 ji).
Calculation:
From the above absorbances, calculate the percent propargite
as follows:
7 = (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
-------
November 1975
Propargite EPA-2
(Tentative)
Determination of |Propargite
by Gas-Liquid Chtlomatography
(TCD - Internal Standard)
Propargite is a common name for 2-(p-tert-butylphenoxy)cyclohexyl-
2-propynyl sulfite, a registered acaricide having the chemical structure:
CH3-
•o-c
HC = C—
CH2-
0
r
-CH2
CH2—0—S=0
Molecular formula: C _H-,0,S
Molecular weight: 350
Melting or boiling point: (not available)
Physical state and color: light t
dark amber viscous liquid of d
25
1.085-1L115; the technical product is at
least 8D%
Solubility: practically insoluble) in water; soluble in most organic
solvents
Stability: (not available)
Other names: Omite, D014 (Uniroyai); Comite
-------
2 Propargite EPA-2
(Tentative)
Reagents:
1. Propargite standard of known % purity
2. Dieldrin standard of known HEOD content
3. Chloroform, pesticide or spectro grade
4. Internal Standard solution - weigh 0.5 gram HEOD into a 25 ml
volumetric flask; dissolve in and make to volume with chloro-
form, (cone 20 mg HEOD/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 4' x 1/4" OD glass, packed with 3% XE-60 on 60/80 mesh
Chromosorb G DMCS (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Usual laboratory glassware
Operating Condj.tions for TCD:
Column temperature: 220°C
Injection temperature: 250°C
Detector temperature: 250°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas pressure: 40 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Propargite EPA-2
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.2 gram propargite standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 10 ml of the internal
standard solution and shake to dissolve, (final cone 20 mg prop-
argite and 20 mg HEOD/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.2 gram propargite
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 10 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the propargite. For
coarse or granular materials, shake mechanically for 30 minutes
or shake by hand intermittently for one hour. (final cone 20 mg
propargite and 20 mg HEOD/ml)
Determination:
Inject 5 ul of standard and, if necessary, adjust the instru-
ment parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is HEOD, then propargite.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order,
Calculation;
Measure the peak heights or areas of propargite and HEOD from
both the standard-internal standard solution and the sample-internal
standard solution.
-------
Propargite EPA-2
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
RF = (wt._HEOp)(% purity HEOD)(pk. ht. or area propargite)
(wt. propargite)(% purity propargite)(pk. ht. or area HEOD)
Determine the percent propargite for each injection of the
sample-internal standard solution as follows and calculate the
average:
7 = (wt. HEOD)(% purity HEOD)(pk. ht. or area propargite) (4-0£>-
' " (wt. sample)(pk. ht. or area HEOD)(RF) '~l
Method submitted by Stelios Gerazounis, EPA, Region II, New York, N. Y.
-------
September 1975 Pyrazon EPA-1
(Tentative)
Determination of Pyrazon in Wettable Powder
by Infrared Spectroscopy
Pyrazon is the accepted common name for 5-amino-4-chloro-2-
phenyl-3(2H)-pyridazinone, a registered herbicide having the
chemical structure:
Molecular formula: C..H.C1N 0
1U o j
Molecular weight: 221.6
Melting point: 207°C with decomposition
Physical state, color, and odor: yellowish-tan to brown powder,
odorless when pure
Solubility: 400 ppm in water at 20°C, 2.8% in acetone, 3.4% in
metHanoi, 0.07% in benzene and in ether, 0.21% in
chloroform, 0.6% in ethyl acetate
Stability: stable; non-corrosive; decomposes at mp
Other names: Pyramin (Badische Anilin-& Soda-Fabrik AG, West Germany)
PGA, H119
Reagents;
1. Pyrazon standard of known % purity
2. Acetonitrile, pesticide or spectro grade
3. Sodium Sulfate, anhydrous, granular
-------
Pyrazon EPA-1
(Tentative)
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.5 mm KBr x>r NaCl cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.08 gram pyrazon standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml acetonitrile by pipette,
close tightly, and shake to dissolve. Add a small amount of
anhydrous sodium sulfate to insure dryness. (final cone 8 mg/ml)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.4 gram pyrazon into
a glass-stoppered flask or screw-cap bottle. Add 50 ml aceto-
nitrile by pipette and 1-2 grams anhydrous sodium sulfate. Close
tightly and shake for one hour. Allow to settle; centrifuge or
filter,taking precaution, to prevent evaporation, (final cone
8 mg pyrazon/ml)
De terminat ion;
With acetonitrile in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 910 cm to
770 cm"1 (11.0 u to 13.0 u).
Determine the absorbance of the standard and sample using
the peak at 826 cm" (12.10 u) and baseline from 844 cm~ to
797 cm"1 (11.85 u to 12.55 u).
-------
Pyrazon EPA-1
(Tentative)
Calculation:
From the above absorbances and using the standard and
sample solution concentrations, calculate the percent pyrazon
as follows:
„ ^ (abs. sample)(cone, std inmg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method contributed by Eva Santos, EPA Region IX, San Francisco,
California.
-------
January 1976 Pyrethrins EPA-1
Description, Structure, Technical Data
The name pyrethrin refers to a registered insecticide consisting
of pyrethrin I and pyrethrin II.
Pyrethrins is the trivial name given to the botanical insecticide
obtained from Chrysanthemum cinerariaefolium. The flowers are the
source of the active principles which are pyrethrin I and II, cinerin
I and II, and jasmolin I and II. The pyrethrin content of flowers and
extracts is as follows: dried flowers 1-3%, crude extract or oleoresin
30-35%, and the most refined grade (dewaxed and decolored) about 60%.
The chemical structure of these compounds is as follows:
R R1
pyrethrin I (C01H.Q0.) -CH, -CH -CH=CH-CH=CH
/I 2.O J J i
pyrethrin II (C00H_00_) -CO-O-CH. -CH.-CH=CH-CH=CH
LL /O J J if
cinerin I (C0_HOQ0_) -CH_ -CH0-CH=CH-CH_
i.\) Zo J j 2. j
cinerin II (C,.,H,..Q0_) -CO-O-CH. -CH0-CH=CH-CH-
£1 lo 5 3 Z J
jasmolin I (C21H3Q03) -CH3 -C
jasmolin II (C22H3o(V -CO-0-CH3 -CH -
-------
2 Pyrethrin EPA-1
Pyrethrin I, cinerin I, and jasmolin I are esters of chrysanthemum
monocarboxylic acid and three different ketonic alcohols; pyrethrin II,
cinerin II, and jasmolin II are esters of chrysanthemum djicarboxylic
acid and the same three alcohols.
Since analysis is based on the isolation and quantitative estima-
tion of the chrysanthemum mono- and di- carboxylic acids, only the total
and not the individual pyrethrins, cinerins, and jasmolins are determined.
However, by convention the total "mono-" acids are reported as "pyrethrin
I" and the total "di-" acids as "pyrethrin II."
Pyrethrins are viscous liquids, practically insoluble in water, but
soluble in alcohol, petroleum ether, kerosene, carbon tetrachloride,
ethylene dichloride, nitromethane, and acetone. They are stable in
water-base aerosols where modern emulsifiers give neutral water systems.
Pyrethrins are oxidized rapidly and become inactive. Stored flowers may
lose 20% of their activity in a year. Impregnated and stabilized dusts
are less susceptible to oxidation than dusts made from ground flowers.
Oxidation is not a problem in stabilized oil concentrates. Antioxidants
such as hydroquinone, pyrogallol, etc. can be used to inhibit oxidation.
Pyrethrins are incompatible with lime and ordinary soaps because acids
and alkalis speed the process of hydrolysis.
Because of its low order of toxicity to warm-blooded animals,
pyrethrin extracts are used extensively in stock sprays, pet sprays,
household sprays and aerosols, industrial sanitation sprays, and to
protect stored food in warehouses.
-------
3 Pyrethrins EPA-1
The use of a synergist, such as piperonyl butoxide, increases the
effectiveness of pyrethrin formulations, enabling the user to maintain
rapid action against insects and to reduce costs.
Pyrethrin formulations available include: concentrated oil
extracts, impregnated and stabilized dusts, and dilute dusts made from
ground flowers. A low color 20% extract in oil has recently become
the "standard" item of the industry.
-------
February 1976 Pyrethrins EPA-2
Determination of Pyrethrins in Formulations
by Gas-Liquid Chromatography (FID)
For description, structure, and technical data on pyrethrins, see
Pyrethrins EPA-1.
Principle of the Method:
The active ingredients in some commercial mixtures of pyrethrins,
piperonyl butoxide (PBO) and n-octylbicycloheptenedicarboximide (NOBD),
especially when present in small amounts (in the range of 0.05-0.50
percent pyrethrin concentrations) can be measured simultaneously by
gas chromatography. A Florisil cleanup procedure is used with all
samples to remove oil-based materials and other substances that would
interfere with the GC analysis of the NOBD compound of the formulation.
Reagents:
1. Pyrethrin primary standard, or extract of known assay
2. Piperonyl butoxide standard of known assay
3. n-Octylbicycloheptenedicarboximide standard of known assay
4. Sodium sulfate, anhydrous
5. Florisil, 60-80 mesh heated for 16 hours at 130° prior to use
6. Hexane, ACS
7. Acetone, ACS
8. Carbon disulfide, ACS
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 51 x 1/8" ID borosilicate glass, packed with 5%
SE-30 on 60-80 mesh Chromosorb W AW DMCS
-------
2 Pyrethrins EPA-2
3. Chromatographic column for Florisil cleanup - 20 mm x 400 mm
borosilicate glass with Ultramax stopcock and 300 ml reservoir
4. Precision liquid syringe: 10 ul
5. liechanical shaker
6. Centrifuge or filtration equipment
7. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 190°C
Injection temperature: 205°C
Detector temperature: 205°C
Carrier gas: Nitrogen
Carrier gas flov? rate: 25 ml/min
Hydrogen flow rate: 25 ml/min
Air flow rate: 200 ml/min
Chart speed: 0.5 in/min
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
Procedure:
Preparation of Standard;
Prepare a mixed standard in carbon disulfide to contain 0.4
ug/jil for pyrethrin I and 1.1 pg/pl each for PBO and NOBD. This
mixed standard is used to quantitate these components in the sample,
Separate standards should be made to identify the individual peaks.
-------
3 Pyrethrins EPA-2
The linearity range for pyrethrin I is 0.2 to 2.2 pg, for PBO
0.6 to 5.6 jag, and for NOBD 0.3 to 1.7 ug, with a minimum detect-
ability of about 0.06 ug for each of the three components.
Preparation of Sample:
A chromatographic column is packed with 5 grams anhydrous
sodium sulfate, followed by 20 grams Florisil, and topped with 5
grams anhydrous sodium sulfate. The column is prewashed with
100 ml hexane, leaving enough solvent in the column to just cover
the packing.
An appropriate weight of sample or sample extract (10 mg
pyrethrin I for a final volume of 25 ml - 0.4 ug/ul) is trans-
ferred to the column with 5-10 ml hexane. The column is washed
with 75 ml hexane and the eluate discarded. The pyrethrin and
the synergistic compounds are then eluted from the column with
125 ml acetone.
The acetone eluate is evaporated nearly to dryness using a
stream of air and a warm steam bath. The residue is diluted to
about 10 ml with carbon disulfide and passed through a small
column of anhydrous sodium sulfate. The sodium sulfate is washed
with a small amount of carbon disulfide, and the combined eluates
are made to a definite volume for chromatographic analysis.
GC Determination:
Using the appropriate attenuation settings, 2 or 3 pi of
standard and sample are alternately injected for pyrethrin 1 and
PBO. Smaller amounts or an additional dilution is needed to keep
the NOBD within the linear range (0.3-1.7 pg for NOBD).
The pyrethrin I component of the pyrethrum fraction of the
formulation is the only predominant peak of the pyrethrum fraction
appearing in the chromatogram under the conditions of this method.
-------
Pvrethrins EPA-2
Other pyrethrum components do not interfere with the simultaneous
recording of the NOBD and PBO components of the mixture.
Calculations:
Use an average of at least three injections of standard and
sample to determine the peak height of each component.
„, (peak ht. sample) (cone. std)(ul std injected) (100)
/, component = -7^—,—: rr/—^ • ;•; 1 r^—:— ,N
(peak ht. std)(cone, sample)(pi sample injected)
The amount of pyrethrin I calculated is multiplied by a
factor of two since pyrethrin I and II usually occur in approx-
imately equal amounts in formulations.
This method is based on "Analytical Studies of Pyrethrin Formulations
ii
by Gas Chromatography by A. Bevenue, Y. Kawano, and F. DeLano, Journal of
Chromatography, 50 (1970), 49-58 and "Analytical Methods for Pesticides
and Plant Growth Regulators, edited by Gunter Zweig, Vol. 6 Gas Chromato-
graphic Analysis, pages 461-464.
-------
January 1976 Pyrethrins EPA-3
Determination of Pyrethrins I & II by Hydrolysis,
Steam Distillation, and Titration (Seil Method)
For definition, structure, and technical data on pyrethrins, see
Pyrethrins EPA-1.
Principle of the Method:
The pyrethrins are hydrolyzed with alcoholic sodium hydroxide to
release the mono- and di- carboxylic acids which together are extracted
with ether and steam-distilled for separation. The tnonocarboxylic acid
"pyrethrin I" is extracted from the distillate while the dicarboxylic
acid "pyrethrin II" is extracted from the residue. Both are titrated
with standard alkali.
Reagents:
1. Petroleum ether, ACS
2. Ethanolic sodium hydroxide solution, 0.5N in ethyl alcohol
3. Barium chloride solution, 10% w/v
4. Phenolphthalein indicator solution, 0.5% in 50% alcohol
5. Sulfuric acid solution, IN
6. Neutral petroleum ether - neutralize with 0.02N NaOH to
faint phenolphthalein pink
7. Standard sodium hydroxide solution, 0.02N
8. Concentrated hydrochloric acid
9. Sodium chloride, ACS
10. Ethyl ether, ACS
-------
2 Pyrethrins EPA-3
Equipment:
1. Soxhlet extraction apparatus
2. Extraction thimbles and cotton or glass wool
3. Dry ice chamber (for aerosols)
4. Water bath
5. Steam bath
6. Reflux apparatus
7. Steam distillation apparatus
Any standard steam distillation apparatus can be used if
the flow of steam and the amount of heat to the distilling
flask can be adjusted so that the volume in the flask remains
constant for most of the distillation but can be reduced to
about 20 ml at the end.
A picture and description of a steam distillation
apparatus is on pages 312-313 of the AOAC 12th Ed. 1975,18.046
and Fig. 18:02.
8. Filter-cell
9. Filtration apparatus
10. Gooch crucible
11. Titration apparatus
12. Usual laboratory glassware
Procedure;
Preparation of Sample:
For solutions, sprays, extracts, and concentrates - Weigh an
amount of sample equivalent to 0.2 gram total pyrethrins into a
250 ml Erlenmeyer flask.
-------
3 Pyrethrins EPA-3
For dusts, powders, flowers, and mosquito coils - Weigh an
amount of sample (finely ground or pulverized if necessary)
equivalent to 0.2 gram total pyrethrins into a Soxhlet thimble,
plug with cotton or glass wool,and place in the Soxhlet extractor.
Add 125 ml petroleum ether and a few boiling chips to a 250 ml
flask and connect to the Soxhlet. Reflux for 6-8 hours. Evaporate
the ether to about 40 ml, stopper the flask, and place in a refrig-
erator at 0-5°C for several hours, preferably overnight. Place a
piece of cotton in the stem of a glass funnel, wet the cotton with
cold petroleum ether, and filter the cold extract, collecting the
filtrate in a 250 ml Erlenmeyer flask. Wash flask several times
with cold ether using a rubber policeman to dislodge any resinous
material in the flask. Add several small glass beads and evap-
orate the ether on a water bath until just less than 1 ml remains.
Do not attempt to remove the last trace of solvent.
For aerosols - Place weighed aerosol can in a dry ice chamber
until well chilled (at least 30 minutes). Punch several holes in
the top of the can and allow the contents to warm slowly to room
temperature. Cut the can open and heat gently on steam bath until
the propellant and other volatile substances are removed so that
the sample can be handled at room temperature without further loss.
Cool, weigh, and transfer the "non-volatile" portion to a bottle.
Rinse the can with ether, dry, and weigh. Calculate percent non-
volatile. Weigh a portion of the non-volatile equivalent to 0.2
gram total pyrethrins into a 250 ml Erlenmeyer flask.
(.2 gram pyrethrins) (% non-volatile) ,. i .. • i j j
°-——~£— —^.—;—r\~—~~ = grams of non-volatile needed
(% claim on label)
„ .. ., (wt. can & contents after heating)-(wt. empty can)
/£ non-volatile = ~, ^—r~; \—T~~ c——
(wt. full can)-(wt. empty can)
-------
Pyrethrins EPA-3
Hydrolysis and Steam Distillation
Add 15 ml of 0.5N ethanolic sodium hydroxide solution to the
sample in the Erlenmeyer flask and reflux for 1 hour. It may be
necessary to add extra 0.5N ethanolic NaOH solution (up to 50 ml)
with samples containing much perfume or other saponifiable
ingredients. Transfer to a large beaker (600-800 ml); wash the
flask with two 25 ml portions of water, adding them to the contents
of the beaker. Add 1 ml deodorized kerosene and dilute to about
200 ml. Place a few glass beads or a boiling tube in the beaker
and boil until the volume is reduced to about 150 ml. If more
than 15 ml of ethanolic. NaOH solution has been used, sufficient
water must be added to insure that all the ethanol is removed
when the volume is reduced to 150 ml. Add 1 gram filter-eel and
transfer the mixture quantitatively to a 250 ml volumetric flask.
(It is more convenient to add the filter-eel to the dry flask first.)
Add 10 ml 10% barium chloride solution, make to volume with water,
and mix thoroughly. Filter through fluted filter paper.
Measure exactly 200 ml of the clear filtrate and transfer
quantitatively to the 500 ml distilling flask of a steam distilla-
tion apparatus. Add one drop of phenolphthalein solution, neutra-
lize with IN sulfuric acid solution,and add 1 ml in excess. Connect
to the steam distillation apparatus and, using a 500 ml separatory
funnel to collect the distillate, steam distill until the volume
remaining in the flask is about 20 ml. The volume of distillate
should be 250-350 ml.
Use the distillate for the determination of Pyrethrin I and the
residue for the determination of Pyrethrin II.
Determination of Pyrethrin I
Add 50 ml neutral petroleum ether to the separatory funnel
containing the distillate and shake thoroughly for one minute.
(If an emulsion forms, add a few crystals of sodium chloride and
-------
5 Pyrethrins EPA-3
shake again.) After the liquids have separated, draw off the
aqueous layer into a second 500 ml separatory funnel to which has
been added a second 50 ml of neutral petroleum ether. Shake for
1 minute and allow to separate, then discard the aqueous layer.
Wash the petroleum ether in the first separatory funnel by shaking
with 10 ral water; using the same 10 ml water, wash the petroleum
ether in the second separatory funnel. Repeat the washing pro-
cedure with a second 10 ml portion of water. Combine the petroleum
ether extracts. Neutralize 15 ml water containing one drop of
phenolphthalein indicator solution with 0.02N sodium hydroxide
solution and add it to the combined petroleum ether extracts.
Titrate with small portions of the 0.02N NaOH solution, shaking
thoroughly after each addition, until the aqueous layer obtains a
pale but permanent pink.
Calculation: The milliequivalent weight of pyrethrin I is
0.3284.
,3284)(100)
v *.u • (ml O.Q2N NaOH)(N 0.02N NaOH)(.:
/. pyrethrxn = (grams sample) (200/250)
Determination of Pyrethrin II
Cool the flask containing the residue from the steam distillation
and filter the solution through a Gooch crucible. Wash the flask
with three 10 ml portions of water using each successively to wash
the Gooch crucible. Transfer the filtrate to a 500 ml separatory
funnel, add 5 ml concentrated hydrochloric acid, and saturate with
sodium chloride. (Acidified aqueous layer must contain visible NaCl
crystals throughout the following extractions.)
Extract the mixture with 50 ml ethyl ether, shaking thoroughly
for one minute. Draw off the aqueous layer into a second separatory
funnel and extract again with 50 ml ethyl ether. Repeat for a third
and fourth extraction using 25 ml ethyl ether each time. Wash the
ether extracts successively with two 10 ml portions of distilled
-------
6 Pyrethrins EPA-3
water. Combine the ether solutions, draw off any water that sep-
arates, and filter through a plug of cotton (previously wetted
with ether) into a 300 ml Erlenmeyer flask. Wash the separatory
funnel and cotton with 10 ml ether. Evaporate the ether on a
water bath and dry the residue at 100°C for 10 minutes. Blow
gently into the flask several times to remove vapors.
Add 30 ml distilled water, boil to dissolve the residue, and
cool. Add a drop of phenolphthalein indicator and titrate with
0.02N NaOH solution to the first pale but permanent pink.
Calculation: The railliequivalent weight of pyrethrin II is
0.1862.
TT - (ml 0.02N NaOH) (N 0.02N NaOH) (.1862) (100)
n ij- - (grams sample) (200/250)
-------
January 1976
Quaternary Ammonium Compounds EPA-1
Definition, Structure, Technical Data,
Halogen and Nitrogen Conversion Factors
A quaternary ammonium compound is an organic nitrogen compound in
which the molecular structure consists of a central pentavalent nitrogen
atom joined to four organic groups and an acidic or basic radical. The
most usual or common of these compounds are salts of mineral acids.
Two examples of the chemical structure are:
(1) a relatively simple salt - alkyl trimethyl ammonium chloride
CH3
H fsj CH3
where "R" represents a long hydrocarbon chain of the length
found in the various fatty acids in which these "quaternaries"
have their origin.
(2) a relatively complex salt - di-isobutyl cresoxy ethoxy ethyl
dimethyl benzyl ammonium chloride, monohydrate
CH3
CH3-C-CH2-C
3 I I
CH3 CH3
The great number of quaternary compounds possible becomes apparent
when consideration is given to the many different organic radicals that
can be attached to the nitrogen, and to the many inorganic radicals that
can form salts.
-------
Quaternary Ammonium Compounds EPA-1
One popular type of quaternary is the water-soluble type which
contains a long carbon chain radical similar to the carbon chain found
in fatty acids. This long chain (alkyl) group imparts surface activity.
In addition to the usual quaternaries, some pentavalent nitrogen
ring compounds such as lauryl pyridinium chloride (structure below) are
also considered quaternary ammonium compounds.
-1+
CH-
-(CH2)
II
•N.
Cl
Most quaternary salts are water-soluble or water-dispersible, but
depending on structure, some are oil-soluble. Many are cationic in
character and are not compatible with soap, anionic wetting agents, or
synthetic detergents.
Quaternary ammonium compounds have many different uses. In the
general field of pesticides, such uses are as disinfectants, cleansers,
sterilizers, deodorants, emulsion stabilizers, fungicides, and algicides.
-------
3 Quaternary Ammonium Compounds EPA-1
CONVERSION FACTORS FOR VARIOUS QUATERNARY AMMONIUM COMPOUNDS
The tables of conversion factors (pages 4 to 9) are based on the
following atomic weights:
Carbon - 12.011 Hydrogen - 1.008 Oxygen - 16.000
Sulfur - 32.064 Nitrogen - 14.007 Chlorine - 35.453
Bromine - 79.909
Percent halogen in the table refers only to the ionic halogen;
where additional halogen is present in the molecule but not figured
in the factor, they are keyed with (*).
Under the percent halogen column there are several materials that
contain no halogen and another element is listed; these are keyed with
(°).
Percent nitrogen in the table refers only to quaternary nitrogen;
where additional nitrogen is present in the molecule but not figured
in the factor, they are keyed with (').
The list is not complete as to all known quaternary materials but
contains the most frequently occurring quaternaries. If specific com-
pounds are not listed, the class name should be checked; i.e., octa-
decyl dimethyl benzyl ammonium chloride will be found under alkyl
dimethyl benzyl ammonium chloride — 100%-C18.
Finally, group names have in some cases been inverted and should be
checked if a particular compound cannot be found; i.e., alkyl dimethyl
methylnaphthyl ammonium chloride will be found under alkyl dimethyl
naphthylmethyl ammonium chloride.
-------
Quaternary Ammonium Compounds EPA-1
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Quaternary Ammonium Compounds bPA-1
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January 1976 Quaternary Ammonium Compounds EPA-2
Determination of Quaternary Ammonium Compounds
*
Qualitative (Auerbach) Tests
For definition, structure, and technical data on these compounds -
see Quaternary Ammonium Compounds EPA-1.
Principle of the Method:
Bromophenol blue indicator forms a salt with quaternary ammonium
compounds. This salt is soluble in ethylene dichloride and colors it
blue.
Reagents;
1. Sodium carbonate, 10% solution
2. Bromophenol blue, 0.04% solution
3. Ethylene dichloride, reagent grade
Equipment;
1. Glass-stoppered test tube or cylinder
2. Pipettes -1,5, and 10 ml
Procedure:
Transfer a portion of sample equivalent to 1-2 rag quaternary
ammonium compound into a glass-stoppered tube or cylinder. Add 5 ml
10% sodium carbonate solution, 1 ml bromophenol blue solution, and
10 ml ethylene dichloride. Shake steadily for 1 to 2 minutes and allow
the layers to separate.
-------
2 Quaternary Ammonium Compounds EPA-2
A blue color in the ethylene dichloride layer indicates the presence
of a quaternary ammonium compound.
Soaps or anionic detergents, if present, may cause the test to fail.
*
This test is based on Auerbach, Industrial & Engineering Chemistry,
Analytical Edition, Vol. 15, Pg. 492 (1943) and Vol. 16, Pg. 739 (1944).
-------
January 1976 Quaternary Ammonium Compounds EPA-3
Determination of Quaternary Ammonium Compounds
by the Ferricyanide Method
For definition, structure, and technical data on these compounds -
see Quaternary Ammonium Compounds EPA-1.
Principle of the Method:
Excess ferricyanide solution is reacted with the quaternary ammonium
compound to form an insoluble precipitate which is filtered from the
sample solution. The excess ferricyanide in the filtrate is determined
by titration with standard thiosulfate,and the percent quaternary is
calculated from the amount of ferricyanide used.
Reagents:
1. Buffer solution - dissolve 130 grams sodium acetate in
about 400 ml water, add 42 ml acetic acid, and make to 500 ml.
2. Ferricyanide solution - dissolve 6.6 grams potassium ferri-
cyanide in water and make to one liter. (approx. 0.02N)
3. Zinc sulfate solution - dissolve 20 grams zinc sulfate hepta-
hydrate in 180 ml water.
4. Sodium thiosulfate, 0.02N standard solution - dilute 100 ml
0.1N standard sodium thiosulfate to 500 ml.
5. Hydrochloric acid, (1+1)
6. Potassium iodide, ACS, crystals
7. Starch indicator solution
Equipment:
1. Steam bath
2. Filtration apparatus
3. Titration apparatus
4. Usual laboratory glassware
-------
2 Quaternary Ammonium Compounds EPA-3
Procedure:
Weigh a portion of sample equivalent to 0.5 gram quaternary ammonium
compound into a 100 ml volumetric flask and dissolve in about 50 ml water.
If the sample is not readily soluble, warm on a steam bath for about 10
minutes with occasional mixing; cool, and add 5 ml of the buffer solution.
Add exactly, by pipette, 30 ml of the ferricyanide solution, swirling the
flask during the addition. Make to volume with water, mix thoroughly, and
let stand for one-half hour, with occasional mixing.
Filter, discarding the first 10 ml of the filtrate. Pipette 50 ml
of the filtrate into a 300 ml glass-stoppered Erlenmeyer flask, add 10 ml
water, 1-2 grams potassium iodide, and 10 ml (1+1) hydrochloric acid.
Mix well and let stand 2 minutes. Add 10 ml zinc sulfate solution, mix
well, and let stand 2-5 minutes longer.
Titrate with standard 0.02N sodium thiosulfate solution, adding
starch indicator solution near the end of the titration.
Repeat the above procedure exactly, using an identical portion (30 ml)
of ferricvanide solution as was used with the sample. This will serve as
a blank for the reagents and provide a basis for calculation.
Calculate the percent nitrogen and percent quaternary ammonium
compounds as follows:
„,„ _ (Blank ml - Sample ml)(N Na.S_O.J (.0140)(100)
/ori " 2. 2. J
(grams sample)
0.0140 = milliequivalent weight of nitrogen
% Quaternary = % nitrogen X nitrogen to quaternary factor
-------
Quaternary Ammonium Compounds EPA-3
The reactions involved in this method are:
1. Precipitation of quaternary with ferricyanide
3[R.R_R,R,N]X + K-Fe(CN), - ) 3KX + [R.R.R.R. ] -Fe(CN) ,
1234 J o 1234 3 o
2. Reaction of excess ferricyanide with potassium iodide
Excess 2K_Fe(CN), + 2KI - ) 2K.Fe(CN), -f I.
J o ' 4 b I.
3. Removal of K.Fe(CN), by zinc sulfate to speed oxidation of KI
4 o
2K4Fe(CN)6
4. Titration of released iodine by sodium thiosulfate
0 NaS0 + 2NaI
-------
January 1976 Quaternary Ammonium Compounds EPA-4
Determination of Quaternary Ammonium Compounds
by the Epton Titration Method
This method is most applicable to formulations containing 0.1% to
1.0% quaternary ammonium compounds.
For definition, structure, and technical data on these compounds -
see Quaternary Ammonium Compounds EPA-1.
Principle of the Method;
An aqueous solution containing a quaternary ammonium compound (QAC)
is reacted with an excess of anionic detergent (AD) in the presence of
methylene blue and chloroform. The excess AD reacts with methylene
blue to form a salt that is soluble in the chloroform (lower) layer and
colors it blue. Since a QAC and an AD react to form an undissociated
salt, any QAC in the sample reduces the AD by an equivalent amount.
The excess AD is titrated by a standard QAC solution. When all of the
AD has reacted with the QAC. the methylene blue is free to dissolve in
the aqueous (upper) layer. The endpoint is therefore the point of
equal color intensity in the two layers when viewed by diffused, reflected
light.
Reagents:
1. Standard QAC, 0.005M solution - dissolve 0.005 gram molecular
weight (usually 2-2.5 grams) of a pure QAC in water and make
to one liter.
2. Standard AD, 0.005M solution - dissolve 0.005 gram molecular
weight (usually 2-2.5 grams) of a pure AD in water and make
to one liter.
-------
2 Quaternary Ammonium Compounds EPA-4
3. Methylene blue indicator solution - dissolve 50 grams sodium
sulfate (anhydrous), 12 ml sulfuric acid, and 0.03 gram
methylene blue in water and make to one liter.
4. Chloroform
Equipment;
1. Glass-stoppered cylinders, 100 ml (plain without graduation
markings is preferred)
2. Burettes and pipettes
3. Source of diffused light
Procedure:
Preparation of Sample:
For best results, dissolve and/or dilute the sample so that a
10 ml aliquot will contain 0.02-0.04 gram of QAC. (Very low per-
cent products requiring extremely large sample amounts may require
a cylinder larger than 100 ml.)
Determination;
Place the sample aliquot in a 100 ml glass-stoppered cylinder,
add 25 ml methylene blue indicator solution, 15 ml chloroform, and
exactly, by pipette, 25 ml of AD solution. Shake thoroughly and
allow to settle; the blue color should be in the bottom layer,
indicating an excess of AD.
Titrate with standard QAC solution in small amounts, shaking
thoroughly after each addition, and allowing time for the layers to
separate. The rate of separation becomes slower as the endpoint is
approached. When color begins to appear in both layers, add the
standard QAC solutio.n in very small increments. The endpoint is
taken as equal color or equal intensity in both layers when viewed
by reflected diffused light. (Should the endpoint be passed,
-------
Quaternary Ammonium Compounds EPA-4
additional AD solution may be added, and the titration continued;
however, that extra amount must be accounted for in the calculations.)
Repeat the titration using 10 ml water as blank and the same
quantity of AD solution as was used for the sample.
Calculation:
The difference between the volume of QAC solution used for the
blank and that used for the sample is the amount equivalent to the
QAC present in the sample.
% QAC Nitrogen = (Blank ml .- Sample ml)(M)(.0140)(100)
° (grams sample)(any dilution factors)
M = molarity of QAC solution
0.0140 = milliequivalent weight of nitrogen
% QAC = % nitrogen X nitrogen to QAC factor
-------
January 1976 Quaternary Ammonium Compounds EPA-5
Determination of Quaternary Chlorides and Bromides
in Mixed Quaternary Formulations by Potentiometric Titration
For definition, structure, and technical data on these compounds -
see Quaternary Ammonium Compounds EPA-1.
Reagents:
1. Inorganic chloride and bromide salts of known halogen content
2. Nitric acid, (1+1)
3. Barium nitrate, crystals, ACS
4. Silver nitrate, 0.1N standard solution
Equipment:
1. Potentiometric titrimeter equipped with a glass reference
electrode and a silver electrode
2. 25 ml burette
3. Usual laboratory glassware
Procedure;
Standardization of Titrimeter:
Prepare a standard solution of chloride and bromide in the
same ratio as expected in the sample. This solution should contain
approximately one milliequivalent total halides (35 mg chloride or
80 mg bromine) in 10 ml solution.
Pipette 10 ml of the prepared standard halide solution into a
250 ml beaker, add 0.5 ml (1+1) hydrochloric acid, and 0.5 gram
barium nitrate (removes iodate in Volhard titration). Place the
electrodes in the solution and set the potential on the titrimeter
at 0.7 or 0.8 volt. Add O.LN silver nitrate solution in small
-------
Quaternary Ammonium Compounds EPA-5
increments and record the new potential after each addition. The
increments should be smallest when the change in potential is
greatest. Continue the addition of silver nitrate and recording
of potential until 3.5 volts is reached.
Plot a curve of each addition of 0.1N silver nitrate against
each potential reading. The plotted curve will indicate two
inflection points; the first will be the bromide end point, and
the second will be the chloride end point. Record the potential
where each end point occurs.
Sample Titration:
Into a 250 ml beaker weigh a portion of sample equal to 1
milliequivalent of total halides. Dilute to 150-200 ml with dis-
tilled water, add 0.5 ml (1+1) nitric acid, and 0.5 gram barium
nitrate. Test the pH of the solution with methyl red. Adjust the
pH by adding small amounts of nitric acid until the solution is red.
Titrate with 0.1N silver nitrate solution and record the volume
added when each potentiometric end point is reached.
Calculations:
Calculate the percent chloride and/or bromide as follows:
(A-B)(N of AgNO )(.03546)(100)
% Chloride = —
(grams of sample)
(B)(N of AgN03)(.07992)(100)
% Bromide = —
(grams of sample)
A = ml of AgNO_ for second (chloride) end point on titration curve
B = ml of AgNO. for first (bromide) end point on titration curve
-------
-------
September 1975
Resmethrin EPA-1
(Tentative)
Determination of Resmethrin in Aerosol
Formulations by Infrared Spectroscopy
Resmethrin is the common name for (5-benzyl-3-furyl)methyl 2,2-
dimethyl-3-(2-methylpropenyl)cyclopropanecarboxylate (approx. 70%
trans, 307, cis isomers), a registered insecticide having the
chemical structure:
0
•/
•CH2—0—C—CH
>-" VjH
-CH3
Molecular formula: ConH0,00
// zb J
Molecular weight: 338
Melting point: 43 to 48°C
Physical state, color, and odor: waxy off-white to tan solid with a
characteristic chrysanthemate odor
Solubility: insoluble in water; soluble in all common organic solvents
Stability: decomposes fairly rapidly on exposure to air and light;
somewhat more stable than pyrethrins
Other names: Synthrin, Crysan, benzofuraline, NIA 17370, FMC 17370,
NRDC 104, SBP 1382
This method is for 1-2% aerosol formulations in which resmethrin
is the only active ingredient.
F
-------
2 Resmethrin EPA-1
(Tentative)
Reagents:
1. Resmethrin standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm KBr or NaCl cells
2. Freezer or dry-ice chest
3. Warm water bath
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.06 gram resmethrin standard into a 10 ml volumetric
flask; dissolve in and make to volume with carbon disulfide.
Add a small amount anhydrous sodium sulfate to insure dryness.
(final cone 6 mg/ml)
Preparation of Sample;
Record the gross weight of a full aerosol can with cap.
Place the can in a freezer overnight or in a dry-ice chest for
at least 2 hours. Remove can and immediately punch several
holes in the top to relieve pressure. Open the can top with
a can opener and allow to warm to room temperature. Remove
any remaining propellants by placing can in a water bath at
about 35-40°C. Dissolve the residue, transfer to a 100 ml
volumetric flask, and make to volume with carbon disulfide.
-------
3 Resmethrin EPA-1
(Tentative)
Dry the can and weigh with cap. Subtract this weight from
the gross weight to obtain the net weight, which is both the
net contents of the sample and the sample weight. From the
declared percent of resmethrin and the sample weight, calculate
the apparent concentration of resmethrin in the 100 ml volu-
metric flask. Dilute an aliquot of this solution to obtain a
solution of approximately 6 mg/ml. Add a small amount of
anhydrous sodium sulfate to insure dryness.
Determination;
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular 1R
instrument being used, scan both the standard and sample from
1800 cm"1 to 1600 cm"1 (5.6 p to 6.25 ji) .
Determine the absorbance of standard and sample using the
peak at 1720 cm" (5.82 p) and baseline from 1765 cm" to
1660 cm"1 (5.67 ju to 6.02 jj) .
Calculation;
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent resmethrin as
follows:
„ m (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method submitted by Mark Law and Jack Looker, EPA Beltsville Chemistry
Laboratory, TSD, OPP, Beltsville, Maryland.
-------
September 1975
Resmethrin EPA-2
(Tentative)
Determination of Resmethrin in Aerosol
Formulations by Gas-Liquid Chromatography (TCD)
Resmethrin is the common name for (5-benzyl-3-furyl)methyl 2,2-
dimethyl-3-(2-methylpropenyl)cyclopropanecarboxylate (approx. 70%
trans, 30% cis isomers), a registered insecticide having the
chemical structure:
o
II
CH2—0—C—CH
^C—CH —C—CH3
Molecular formula: C22H26°3
Molecular weight: 338
Melting point: A3 to 48°C
Physical state, color, and odor: waxy off-white to tan solid with a
characteristic chrysanthemate odor
Solubility: insoluble in water; soluble in all common organic solvents
Stability: decomposes fairly rapidly on exposure to air and light;
somewhat more stable than pyrethrins
Other names: Synthrin, Crysan, benzofuraline, NIA 17370, FMC 17370,
NRDC 104, SBP 1382
This method is for 1-2% aerosol formulations in which resmethrin
is the only active ingredient.
-------
2 Resmethrin EPA-2
(Tentative)
Reagents;
1. Resmethrin standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
(Methanol could be substituted for the carbon disulfide
in this method if it is desired to use the same sample
solutions for High Pressure Liquid Chromatography —
see EPA-4.)
Equipment;
1. Gas chromatograph with thermal conductivity detector
2. 4' x 1/4" column packed with 10% SP-2100 on Chromosorb 750,
80/100 mesh (or equivalent column)
3. 25 jil precision syringe
4. Freezer or dry-ice chest
5. Warm water bath
6. Usual laboratory glassware
Determination using Thermal Conductivity Detector:
Operating Conditions;
Column temperature: 260°C
Injection temperature: 290°C
Detector temperature: 270°C
Filament current: 200 ma
Carrier gas: Helium
Flow rate: 55 ml/min
Operating conditions for filament current, column temperature,
or gas flow should be adjusted by the analyst to obtain optimum
response and reproducibility.
-------
Resmethrin EPA-2
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.06 gram resmethrin standard into a 10 ml volumetric
flask; dissolve in and make to volume with carbon disulfide.
Add a small amount anhydrous sodium sulfate to insure dryness.
(final cone 6 mg/ml)
Preparation of Sample:
Record the gross weight of a full aerosol can with cap.
Place the can in a freezer overnight or in a dry-ice chest for
at least 2 hours. Remove can and immediately punch several
holes in the top to relieve pressure. Open the can top with
a can opener and allow to warm to room temperature. Remove any
remaining propellants by placing can in a water bath at about
35-40°C. Dissolve the residue, transfer to a 100 ml volumetric
flask, and make to volume with carbon disulfide.
Dry the can and weigh with cap. Subtract this weight from
the gross weight to obtain the net weight, which is both the
net contents of the sample and the sample weight. From the
declared percent of resmethrin and the sample weight, calculate
the apparent concentration of resmethrin in the 100 ml volu-
metric flask. Dilute an aliquot of this solution to obtain a
solution of approximately 6 mg/ml.
Determination:
Using a precision liquid syringe, alternately inject three
10-20 fil portions of standard and sample solutions. Measure the
peak height or peak area for each peak and calculate the average
for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
-------
Resmethrin EPA-2
(Tentative)
Calculation:
From the average peak height or peak area, calculate
the percent resmethrin as follows:
2 . (pk. ht. or area sample)(wt. std injected)(Z purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Mark Law and Jack Looker, EPA-OPP-TSD Beltsville
Chemistry Laboratory, Beltsville, Maryland.
-------
October 1975
Resmethrin EPA-3
(Tentative)
Determination of Resmethrin in Aerosol
Formulations by Gas-Liquid Chromatography
(TCD - Internal Standard)
Resmethrin is the common name for (5-benzyl-3-furyl) methyl 2,2-
dimethyl-3-(2-methylpropenyl) cyclopropanecarboxylate (approx. 70%
trans, 30% cis isomers), a registered insecticide having the
chemical structure:
0
II
CH2—0—C—CH
0'
CH3
—CH=C—CH3
CH3
Molecular formula: C__H 0
2.2. 2o J
Molecular weight: 338
Melting point: 43 to 48°C
Physical state, color, and odor: waxy off-white to tan solid with a
characteristic chrysanthemate odor
Solubility: insoluble in water; soluble in all common organic solvents
Stability: decomposes fairly rapidly on exposure to air and light;
somewhat more stable than pyrethrins
Other names: Synthrin, Crysan, benzofuraline, NIA 17370, FMC 17370,
NRDC 104, SBP 1382
This method is for 1-2% aerosol formulations in which resmethrin
is the only active ingredient.
-------
2 Resmethrin EPA-3
(Tentative)
Reagents:
1. Resmethrin standard of known % purity
2. Dieldrin standard of known HEOD content
3. Benzene, pesticide or spectro grade
4. Internal Standard solution - weigh 0.2 gram dieldrin standard
into a 10 ml volumetric flask; dissolve in and make to volume
with benzene. (cone 20 mg dieldrin/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector
2. 6' x 1/8" stainless steel column packed with 10% SE 30
on 80/100 Diatoport S (or equivalent column)
3. Precision liquid syringe: 10 or 25 pi
4. Freezer or dry-ice chest
5. Warm water bath
6. Usual laboratory glassware
Operating Conditions for TCP:
Column temperature: 230°C
Injection temperature: 250°C
Detector temperature: 250°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas pressure: 25 ml/min
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Resmethrin EPA-3
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.16 gram resmethrin into a 10 ml volumetric flask;
dissolve in and make to volume with benzene. Pipette 5 ml of
this solution and 5 nl internal standard solution into a small
flask or vial and mix thoroughly. (cone 8 mg resmethrin and
10 mg dieldrin/ml)
Preparation of Sample;
Record the gross weight of a full aerosol can with cap.
Place the can in a freezer overnight or in a dry-ice chest for
at least 2 hours. Remove can and immediately punch several
holes in the top to relieve pressure. Open the can top with a
can opener and allow to warm to room temperature. Remove any
remaining propellants by placing can in a water bath at about
35-40°C. Dissolve the residue, transfer to a 100 ml volumetric
flask, and make to volume with benzene.
Dry the can and weigh with cap. Subtract this weight from
the gross weight to obtain the net weight, which is both the
net contents of the sample and the sample weight. From the
declared percent of resmethrin and the sample weight, calculate
the apparent concentration of resmethrin in the 100 ml volu-
metric flask. Dilute an aliquot of this solution to obtain a
concentration of 16 mg/ml. Pipette 5 ml of this diluted solution
and 5 ml internal standard solution into a small flask or vial
and mix thoroughly. (cone 8 mg resmethrin and 10 mg dieldrin/ml)
Determination:
Inject 5-10 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within 10 minutes and peak heights of from 1/2 to 3/4
full scale. The elution time of dieldrin is 3.5 minutes and
that of resmethrin 6.0 minutes.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions.
-------
Resmethrin EPA-3
(Tentative)
Calculation:
Measure the peak heights or areas of resmethrin and dieldrin
from both the standard-internal standard solution and the sample-
internal standard solutions.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. dieldrin)(% purity dieldrin)(pk. ht. or area resmethrin)
(wt. resmethrin)(% purity resmethrin)(pk. ht. or area dieldrin)
Determine the percent resmethrin for each injection of the
sample-internal standard solution as follows and calculate the
average:
% = (wt. dieldrin)(% purity dieldrin)(pk. ht. or area resmethrin)-Q^fr)
(wt. sample)(pk. ht. or area dieldrin)(RF) 6/-/J
Method submitted by Stelios Gerazounis, EPA Region II, New York, New York.
-------
September 1975
Resmethrin EPA-4
(Tentative)
Determination of Resmethrin in Aerosol Formulations
by High Pressure Liquid Chromatography
Resmethrin is the common name for (5-benzyl-3-furyl)methyl 2,2-
dimethyl-3-(2-methylpropenyl)cyclopropanecarboxylate (approx. 70%
trans, 30% cis isomers), a registered insecticide having the chemical
structure:
y—CH2-
-/
0
II
CH2—0—C-
•CH
X
CH3
'C— CH = C— CH3
Molecular formula: C0_H0,CL
ii 2b 3
Molecular weight: 338
Melting point: A3 to 48°C
Physical state, color, and odor: waxy off-white to tan solid with a
characteristic chrysanthemate odor
Solubility: insoluble in water; soluble in all common organic solvents
Stability: decomposes fairly rapidly on exposure to air and light;
somewhat more stable than pyrethrins
Other names: Synthrin, Crysan, benzofuraline, NIA 17370, FMC 17370,
NRDC 104, SBP 1382
This method is for 1-2Z aerosol formulations in which resmethrin
is the only active ingredient.
-------
Resmethrin EPA-4
(Tentative)
Reagents;
1. Resmethrin standard of known % purity
2. Methanol, pesticide or spectre grade
Equipment;
1. High pressure liquid chromatograph
2. High pressure liquid syringe
3. Liquid chromatographic column such as DuPont's ODS
Permaphase 1 meter x 2.1 m I.D. (or equivalent column)
4. Freezer or dry-ice chest
5. Warm water bath
6. Usual laboratory glassware
Operating Conditions for DuPont Model 830;
Mobile phase: 70% methanol + 30% water
Column temperature: 65°C
Column pressure: 1000 psl
Observed flow rate: 1-2 ml/min
Detector: UV at 254 nm
Chart speed: 5 min/in
Injection: 5 pi
Procedure:
Preparation of Standard;
Weigh 0.06 gram resmethrin standard into a 10 ml volumetric
flask; dissolve in and make to volume with methanol. (final
cone 6 mg/ml)
-------
Resmethrin EPA-4
(Tentative)
Preparation of Sample;
Record the gross weight of a full aerosol can with cap.
Place the can in a freezer overnight or in a dry-ice chest for
at least 2 hours. Remove can and immediately punch several
holes in the top to relieve pressure. Open the can top with
a can opener and allow to warm to room temperature. Remove
any remaining propellents by placing can in a water bath at
about 35-40°C. Dissolve the residue, transfer to a 100 ml
volumetric flask, and make to volume with methanol.
Dry the can and weigh with cap. Subtract this weight from
the gross weight to obtain the net weight, which is both the
net contents of the sample and the sample weight. From the
declared percent of resmethrin and the sample weight, calculate
the apparent concentration of resmethrin in the 100 ml volu-
metric flask. Dilute an aliquot of this solution to obtain a
solution of approximately 6 mg/ml.
Determination;
Using a high pressure liquid syringe, alternately inject
three 5 jil portions each of standard and sample solutions.
Measure the peak height or peak area for each peak and calculate
the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation;
From the average peak height or peak area,calculate the
percent resmethrin as follows:
2 . (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer Hayes, EPA-OPP-TSD Beltsville Chemistry
Laboratory, Beltsville, Maryland.
-------
October 1975
Resmethrin EPA-5
(Tentative)
Determination of Resmethrin by
Gas-Liquid Chromatography
(FID - Internal Standard)
Resmethrin is the common name for (5-benzyl-3-furyl) methyl 2,2-
dimethyl-3-(2-methylpropenyl)cyclopropanecarboxylate (approx. 70%
trans, 30% cis isomers), a registered insecticide having the chemical
structure:
CH2—0—
0
II
C—CH
•C—CH=C—CH3
CH3
Molecular formula: C00H0/.00
Z/ /D J
Molecular weight: 338
Melting point: 43 to 48°C
Physical state, color, and odor: waxy off-white to tan solid with a
characteristic chrysanthemate odor
Solubility: insoluble in water; soluble in all common organic solvents
Stability: decomposes fairly rapidly on exposure to air and light;
somewhat more stable than pyrethrins
Other names: Synthrin, Crysan, benzofuraline, NIA 17370, FMC 17370,
NRDC 104, SBP 1382
-------
Resmethrin EPA-5
(Tentative)
Reagents:
1. Resmethrin standard of known % purity
*
2. Dipentyl phthalate, practical
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.1 gram dipentyl
phthalate into a 100 ml volumetric flask; dissolve in
and make to volume with acetone. (cone 1 mg dipentyl
phthalate/ml)
* #P2473 Eastman Catalog #48, Eastman Organic Chemicals,
Rochester, N. Y. 14650
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm ID glass column packed with 5% SE-30
on 80/100 Chromosorb W HP (or equivalent column)
3. Precision liquid syringe: 5 or 10 jil
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 210°
Injection temperature: 260°
Detector temperature: 260°
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi
Hydrogen pressure: 20 psi
Air pressure: 30 psi
-------
3 Resmethrin EPA-5
(Tentative)
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure:
Preparation of Standard:
Weigh 0.05 gram resmethrin standard into a small glass-
stoppered flask or screw-cap tube. Add by pipette 25 ml of the
internal standard solution and shake to dissolve, (final cone
2 mg resmethrin and 1 mg dipentyl phthalate/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.05 gram resmethrin
into a small glass-stoppered flask or screw-cap tube. Add by
pipette 25 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the resmethrin.
For coarse or granular materials, shake or tumble mechanically
for 30 minutes or shake by hand intermittently for one hour.
(final cone 2 mg resmethrin and 1 mg dipentyl phthalate/ml)
Determination:
Inject 1-2 jil of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is dipentyl phthalate, then
resmethrin.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
-------
Resmethrin EPA-5
(Tentative)
Calculation:
Measure the peak heights or areas of resmethrin and dipentyl
phthalate from both the standard-internal standard solution and
the sample-internal standard solutions.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
DPP = dipentyl phthalate = internal standard
(wt. DPP)(% purity DPP)(pk. ht. or area resmethrin)
(wt. resmethrin)(% purity resmethrin)(pk. ht. or area DPP)
Determine the percent resmethrin for each injection of the
sample-internal standard solution as follows and calculate the
average:
(wt..DPP)(% purity DPP)(pk. ht. or area resmethrin)
(wt. sample)(pk. ht. or area DPP)(RF)
This method was submitted by the Commonwealth of Virginia, Division
of Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
Note! This method has been designated as tentative since it is a
Va. Exp. method and because some of the data has been suggested
by EPA's Beltsville Chemistry Lab. Any comments, criticism,
suggestion, data, etc. concerning this method will be appreciated,
-------
August 1975
Ronnel EPA-1
Determination of Ronnel
by Infrared Spectroscopy
Ronnel is the accepted common name for 0,0-dimethyl 0-(2,4,5-
trichlorophenyl) phosphorothioate, a registered insecticide having
the chemical structure:
Cl
Molecular formula: C0H0C100_PS
o o j J
Molecular weight: 321.5
Melting point: softens at 35 to 37°C with full melt at 40-42°C
Physical state and color: white crystalline powder
Solubility: 40 ppm in water at RT; readily soluble in most organic
solvents including refined kerosene
Stability: stable at temperatures to 60°C, and in neutral or acidic
media; hydrolyzed by alkali to the desmethyl compound;
not compatible with alkaline pesticides
Other names: fenchlorphos (common name accepted by ISO and BSI);
Trolene (drug grade) and Korlan (tech. grade)(Dow Chemical
Co.); Nankor, Ectoral, Etrolene, Vlozene
-------
2 Ronnel EPA-1
Reagents:
1. Ronnel standard of known % purity
2. Carbon disulfide, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.5 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.05 gram ronnel into a small glass-stoppered flask
or screw-cap bottle, add 10 ml carbon disulfide by pipette, and
shake to dissolve. Add a small amount of anhydrous sodium
sulfate to insure dryness. (final cone. 5 mg/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.125 gram ronnel
into a glass-stoppered flask or screw-cap bottle. Add 25 ml
carbon disulfide and 1-2 grams anhydrous sodium sulfate. Close
-------
3 Ronnel EPA-1
tightly and shake for one hour. Allow to settle; centrifuge or
filter if necessary, taking precaution to prevent evaporation.
(final cone 5 mg ronnel/ml)
Determination:
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan both the standard and sample from
1020 cm"1 to 890 cm'1 (9.8 p to 11.3 >i).
Determine the absorbances of the standard and sample using
—1 —1
the peak at 960 cm (10.42 u) and basepoint at 920 cm (10.87 ;i).
Calculation;
From the above absorbances and using the standard and sample
.concentrations, calculate the percent ronnel as follows:
<•/ = (abs. sample) (cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg ronnel/ml carbon disulfide gives
an absorbance of approx. 0.08 in a 0.5 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services.
-------
November 1975
Ronnel EPA-2
Determination of Ronnel
by Gas-Liquid Chromatography
(FID - Internal Standard)
Ronnel is the accepted common name for 0,0-dimethyl 0-(2,4,5-
trichlorophenyl) phosphorothioate, a registered insecticide having
the chemical structure:
Cl
CH3 0
CH3 0
Molecular formula: C0H_C1,0,PS
o o 33
Molecular weight: 321.5
Melting point: softens at 35 to 37°C with full melt at 40-42°C
Physical state and color: white crystalline powder
Solubility: 40 ppm in water at RT; readily soluble in most organic
solvents including refined kerosene
Stability: stable at temperatures to 60°C, and in neutral or acidic
media; hydrolyzed by alkali to the desmethyl compound;
not compatible with alkaline pesticides
Other names: fenchlorphos (common name accepted by ISO and BSI);
Trolene (drug grade) and Korlan (tech. grade)(Dow Chemical
Co.); Nankor, Ectoral, Etrolene, Viozene
-------
2 Ronnel EPA-2
Reagents;
1. Ronnel standard of known % purity
2. Diisobutylphthalate
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.07 gram diisobutyl-
phthalate into a 100 ml volumetric flask, dissolve in, and
make to volume with acetone, (cone 0.7 mg/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 4 mm ID glass, packed with 3% OV-1 on 60/80 mesh
Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 jil
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 180°C
Injection temperature: 250°C
Detector temperature: 250°C
Carrier gas: Nitrogen
Carrier gas pressure: (not stated in method)
Hydrogen pressure: 24 psi
Air pressure: 30 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response and
reproducibility.
-------
3 Ronnel EPA-2
Procedure;
Preparation of Standard;
Weigh 0.05 gram ronnel standard into a small glass-stoppered
flask or screw-cap bottle. Add by pipette 25 ml of the internal
standard solution and shake to dissolve, (final cone 2 mg ronnel
and 0.7 mg diisobutylphthalate/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.05 gram ronnel into
a small glass-stoppered flask or screw-cap bottle. Add by pipette
25 ml of the internal standard solution. Close tightly and shake
thoroughly to dissolve and extract the ronnel. For coarse or
granular materials, shake mechanically for 30 minutes or shake
by hand intermittently for one hour, (final cone 2 mg ronnel and
0.7 mg diisobutylphthalate/ml)
Determination;
Inject 3-4 j»l of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is diisobutylphthalate, then
ronnel.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of ronnel and diisobutyl-
phthalate from both the standard-internal standard solution and
the sample-internal standard solution.
-------
4 Ronnel EPA-2
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
I.S. « internal standard « diisobutylphthalate
op = (wt. I.S.)(% purity I.S.)(pk. ht. or area Ronnel)
(wt. Ronnel)(% purity Ronnel)(pk. ht. or area I.S.)
Determine the percent Ronnel for each injection of the sample-
internal standard solution as follows and calculate the average:
% m (wt. I.S.)(% purity I.S.)(pk. ht. or area Ronnel)
(wt. sample) (pk. ht. or area I.S.)(RP)
Method submitted by Division of Regulatory Services, Kentucky Agricul-
tural Experiment Station, University of Kentucky, Lexington, Kentucky 40506.
-------
January 1976
Rotenone EPA-1
Determination of Rotenone
in Pesticides - Qualitative tests
Rotenone is a registered insecticide having the chemical structure:
0-CH3
CH3-0
Molecular formula: C0_H_,,0,
23 a o
Molecular weight: 394.4
Melting point: 163°C (a dimorphoric form melts at 181°C)
Physical state and color: colorless crystals; crystallizes with solvent
of crystallization
Solubility: 15 ppm in water at 100°C; slightly soluble in petroleum
oils, carbon tetrachloride; soluble in polar organic
solvents
Stability: readily oxidized, especially in presence of light or alkali
Other names: Protex, Derris, Lonchocarpus, Barbasco (Spanish-speaking
countries of So. Am.), Cube (Peru), Haiari (British Guiana),
Nekos (Dutch Guiana), Timbo (Brazil), Nicouline, tubatoxin
Reagents;
1. Chloroform, ACS
2. Thymol solution - dissolve 10 grams of thymol in 100 ml of
chloroform.
3. Nitric acid-hydrochloric acid mixture - add 0.2 ml of concen-
trated nitric acid to 100 ml of concentrated hydrochloric acid.
-------
2 Rotenone EPA-1
Equipment;
1. Glass-stoppered test tubes or small flasks
2. Usual laboratory glassware
Preparation of Sample:
Dilute an amount of liquid sample, or extract an amount of dry
sample with chloroform to give 0.01-0.25 mg of rotenone per ml of
solution.
This method is sensitive to 0.01 mg of rotenone per ml, but if
too much rotenone is present the characteristic blue color will not
develop. If the test fails on a sample believed to contain rotenone,
repeat on a diluted portion of the sample.
Qualitative Determination:
Place 5 ml of sample solution, 5 ml of thymol solution, and 3 ml
mixed acid solution in a glass-stoppered test tube or small flask.
Agitate for about 30 seconds and allow to stand.
The presence of rotenone is indicated by the appearance of a bluish-
green to blue color. The color usually appears in from 30 seconds to 2
minutes and deepens on standing.
In the presence of the yellow coloring matter of pyrethrum flowers
and of derris extract, the developed color may be green at first but on
standing will become bluish-green and finally blue.
-------
-------
November 1975
Salicylanilide EPA-1
Determination of Salicylanilide
by Ultraviolet Spectroscopy
Salicylanilide is a registered fungicide having the chemical
structure:
0 H
— N
•OH
Molecular formula: C H NO
Molecular weight: 213.3
Melting point: 135°C
Physical state and color: cream-colored powder
Solubility: almost insoluble in water (55 ppm at 25°), slightly soluble
in organic solvents
Stability: slightly volatile in steam; forms water-soluble salts with
alkali metals, ammonia, amines, and forms insoluble salts
with copper and zinc
Other names: Shirlan (ICI Ltd)
Reagents:
1. Salicylanilide standard of known % purity
2. Sodium hydroxide, 0.1N solution (this need not be standardized)
-------
2 Salicylanilide EPA-1
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Soxhlet extraction apparatus
3. Rotary evaporator or steam bath and compressed air source
4. Usual laboratory glassware
Procedure:
Preparation of Standard:
Weigh 0.1 gram salicylanilide into a 100 ml volumetric flask?
dissolve in and make to volume with O.IK sodium hydroxide solution.
Mix thoroughly, pipette 10 ml into a second 100 ml volumetric flask,
and make to volume with the 0.1N NaOH solution. Again, mix thor-
oughly, and pipette 10 ml into a third 100 ml volumetric flask.
Make to volume with 0.1N NaOH solution and mix well. (final cone
10 pg/ml)
Preparation of Sample:
For salicylanilide formulations, weigh a portion of sample
equivalent to 0.01 gram salicylanilide into a 100 ml volumetric
flask, make to volume with 0.1N sodium hydroxide solution, and mix
thoroughly. Pipette 10 ml into a second 100 ml volumetric flask,
again make to volume with the 0.1N sodium hydroxide solution, and
mix well. (final cone 10 pg salicylanilide/ml)
For salicylanilide-treated products weigh a portion of sample
equivalent to 0.01 gram salicylanilide into a Soxhlet thimble, plug
with cotton or glass wool, and extract with ethanol for about two
hours. Evaporate to dryness using a rotary evaporator or a steam
Salicylanilide is used to prevent mildew on such things as rope,
canvass, upholstery and mattress filling, tiles, in rubber backing
(0.5%) for carpets and carpet underlays.
-------
3 Salicylanilide EPA-1
bath with a gentle stream of air. Dissolve residue, transfer to a 100
ml volumetric flask, and make to volume with O.lN sodium hydroxide
solution, and mix thoroughly. Pipette 10 ml into a second 101 mi
volumetric flask; make to volume with the O.lN sodium hydroxide
solution. (final cone 10 jig salicylanilide/ml)
UV Peterminatipn:
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 338 nm with O.lN
sodium hydroxide solution in each cell. Scan both the standard and
sample from 360 nm to 250 nm vith O.lN NaOH solution in the reference
cell. Measure the absorbance of both standard and sample at 338 nm.
If an untreated product is available, it can be carried through
the extraction procedure and used as a blanl:. The absorbance at
338 nm would then be subtracted from the sample absorbance at 338 nm-
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent Salicylanilide as follows:
a. _ (abs. sample) (cone, std in pg/ml)(% purity std)
(abs. std) (cone, sample in jjg/ml)
-------
September 1975 Siduron EPA-1
(Tentative)
Determination of Siduron
by Ultraviolet Spectroscopy
Siduron is the accepted common name for l-(2-methylcyclohexyl)-
3-phenylur ea , a registered herbicide having the chemical structure:
CH3
CH - CH2
H 0 H / \
I II I / \
N— C— N— CH CH2
— CH2
Molecular formula: CiA^n1*?0
Molecular weight: 232.3
Melting point: 133 to 138°C
Physical state, color, and odor: odorless, white, crystalline solid
Solubility: 18 ppm in water at 25°C; soluble to the extent of 10% or
more in cellosolve, dimethylacetamide, dime thy Iformamide,
ethanol, isophorone, methylene chloride
Stability: stable up to its m.p. in water; slowly decomposed by
acids and bases; non-corrosive
Other names: Tupersan (DuPont)
Reagents :
1. Siduron standard of known % purity
2. Methanol, pesticide or spectro grade
-------
Siduron EPA-1
(Tentative)
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm silica cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard;
Weigh 0.1 gram siduron standard into a 100 ml volumetric
flask, add 100 ml methanol by pipette, and mix thoroughly.
Pipette 10 ml into a second 100 ml volumetric flask, make to
volume with methanol, and mix thoroughly. Pipette 5 ml into
a third 100 ml volumetric flask, make to volume with methanol,
and mix thoroughly, (final cone 5
Preparation of Sample:
Weigh a portion of sample equivalent to 0.1 gram siduron
into a 250 ml glass-stoppered or screw-cap flask, add 100 ml
methanol by pipette, and shake on a mechanical shaker for 30
minutes. Allow to settle; centrifuge or filter if necessary,
taking precautions to prevent evaporation. Pipette 10 ml into
a 100 ml volumetric flask, make to volume with methanol, and
mix thoroughly. Pipette 5 ml of this solution into another
100 ml volumetric flask, make to volume with methanol, and mix
thoroughly, (final cone 5 jig siduron/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen at 0 and 100% transmission at 240 nm with
-------
Siduron EPA-1
(Tentative)
methanol in each cell. Scan both the standard and sample from
300 nm to 200 nm with methanol in the reference cell.
Measure the absorbance of standard and sample at 240 nm.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent siduron as follows:
7 . (abs. sample)(cone. std in pg/ml)(% purity std)
(abs. std) (cone, sample in ug/ml)
Method submitted by Stelios Gerazounis, EPA Region II, New York, N. Y.
-------
September 1975
Simazine EPA-1
(Tentative)
Determination of Simazine in 0.1% Aqueous
Suspension by Ultraviolet Spectroscopy
Simazine is the accepted common name for 2-chloro-4,6-bis
(ethylamino)-s-triazine, a registered herbicide having the
chemical structure:
Cl
N
—CH2 —N C
H
C —N —CH2—CH3
Molecular formula: C7H -C1N
Molecular weight: 201.7
Melting point: 225 to 227eC
Physical state and color: white, crystalline solid
*
Solubility: at 20°C, 2 ppm in petroleum ether, 5 ppm in water,
400 ppm in methanol, and 900 ppm in chloroform;
considered slightly soluble in chloroform, dioxane,
and ethylcellosolve
Stability: stable in neutral or slightly acidic or basic media;
hydrolyzed by stronger acids and bases, especially at
higher temperatures; non-corrosive
Other names: Princep, Gesatop, Primatol, and Printop (CIBA-GEIGY);
Simanex
-------
2 Slmazine EPA-1
(Tentative)
This method is designed specifically for 0.1% aqueous suspensions;
however, it may be used for other simazine formulations with appro-
priate modifications when there is no interference at the 263 rap.
maxima.
Reagents:
1. Simazine standard of known purity
<»
2. Methanol - ACS
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm silica cells
2. Steam bath
3. Flow of dry, clean air
4. Usual laboratory glassware
Procedurei
Preparation of Standard:
Weigh 0.05 gram of simazine standard into a 100 ml volumetric
flask; dissolve and make to volume with methanoL Mix thoroughly.
Pipette 5 ml into a second 100 ml volumetric flask, make to
volume, and mix well, (final cone 25 fig/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.0025 gram simazine
(2.5 g of 0.1% formulation) from a weighing buret into a 50 ml
beaker and take to dryness on a steam bath with a current of
clean, dry air. Transfer the residue to a 100 ml volumetric
-------
3 Simazine EPA-1
(Tentative)
flask with small portions of methanol, make to volume with
methanol, and mix thoroughly. Filter through Whatman No. 5 just
prior to UV determination, (final cone 25 ug simazine/ml)
UV Determination;
Using the optimum quantitative settings for the particular
UV instrument being used, adjust the 0 and 100% settings at
263 mp with methanol in both cells. Scan both standard and
sample from 360 mp to 230 mu.
Calculation;
Measure the absorbance (A) of both standard and sample at
263 mu (maxima) and 300 mu (base point) . Calculate the percent
of simazine as follows:
samPleMconc' std in ug/ml)(% purity std)
-, 3~A3QQ std) (cone, sample in ug/ml)
The absorbance is linear for the concentration range of
0-50 ^ig/ml in methanol.
Method submitted by Dean Hill, EPA Region IX, San Francisco, California.
-------
December 1975
Sodium Chlorate EPA-1
Determination of Sodium Chlorate in
Herbicides by Reduction and Titration
Sodium chlorate is a registered herbicide, having the chemical
structure:
0=01=0
0
Molecular formula:
Molecular weight:
Melting point:
NaCIO
106. A
248°C; decomposes about 300°C with evolution of
oxygen
Physical state, color, odor, and taste: white to pale yellow, odorless
crystals with a salty taste
Solubility: soluble in water 79 g/100 ml at 0°C and 230 g/100 ml at
100eC; somewhat soluble in alcohol and glycerol
Stability: DANGEROUSLY FLAMMABLE!; strong oxidizing agent, hence
serious fire hazard with organic matter, e.g., vegetation,
clothing, shoes (easily ignited by friction or heat as on
shoestrings or cloth apron strings); DO NOT BURN contam-
inated clothing or containers. Somewhat corrosive to zinc
and mild steel
Other names: Atlacide, Atratole, De-Fol-Ate, Drop-Leaf, Klorex, Fall,
Rasikal, Shed-a-Leaf
Principle of the Method;
The sodium chlorate in a portion of sample is reacted (reduced)
with a known amount (in excess) of ferrous sulfate solution. The
ferrous sulfate not used by the sodium chlorate is titrated with standard
potassium permanganate solution. An identical amount of ferrous sulfate
solution without sample is titrated and the difference used to calculate
the sodium chlorate in the sample.
-------
2 Sodium Chlorate EPA-1
Reagents;
1. Potassium permanganate, 0.1N standard solution
2. Ferrous sulfate solution - dissolve 30 grams of ferrous sulfate
heptahydrate (FeSO,.7H 0) in 900 ml water and make to one liter
with concentrated sulfuric acid.
3. Manganese sulfate solution - weigh 14 grams of manganous sulfate
tetrahydrate (MnSO,.4H20) into 200 ml volumetric flask, add 25 ml
sulfuric acid and 25 ml 857« phosphoric acid, and make to volume
with water.
Equipment;
1.
300 ml Erlenmeyer flask with rubber stopper fitted with a
Bunsen valve (described below)
-B
•A
D
•C
2.
3.
4.
5.
The Bunsen valve is a short 2-4"
length of rubber tubing (A) stoppered
at one end (B) and fitted over a piece
of glass tubing (C) at the other end.
A 1/2-3/4" slit (D) is made with a
razor blade along the length of the
tubing. This slit allows internal
pressure to be relieved by allowing
gases to escape, but is sealed as out-
side pressure pushes in since the sides
of the slit are pressed together.
Mechanical shaker
Filtration apparatus
Titration apparatus
Usual laboratory glassware
-------
3 Sodium Chlorate EPA-1
Procedure:
Weigh a portion of sample equivalent to 0.6 gram sodium chlorate into
a 500 ml glass-stoppered or screw-cap flask, add exactly 250 ml water,
shake on a mechanical shaker for two hours, and filter. Pipette a 25 ml
aliquot into a 300 ml Erlenmeyer flask, add by pipette 30 ml ferrous
sulfate solution, close tightly with a rubber stopper fitted with a
Bunsen valve (to prevent oxidation by air), and boil 10 minutes.
Cool, dilute to about 100 ml with water, add 10 ml of the manganese
sulfate solution, and mix well. Titrate with 0.1N potassium permanganate
solution to the first distinct pink color. The endpoint is not permanent
due to the oxidation of the chloride by the permanganate.
Repeat the same procedure using an identical 30 ml portion of the
ferrous sulfate solution but no sample solution. The difference between
these two titrations in ml of 0.1N potassium permanganate represents the
sodium chlorate in the aliquot of sample solution.
Calculation:
From the difference in titration, calculate the percent sodium
chlorate as follows:
net ml 0.1N KMnO, « ml used for FeSO, alone - ml used for FeSO, and sample
(net ml KMnO.)(N KMnO. ) (0.01774) (100)
_ « 4_ 4
(gram sample)(25/250)
0.01774 « milliequivalent weight of NaClO.
-------
January 1976
Streptomycin EPA-1
Determination of Streptomycin by
Ultraviolet or Colorimetric Spectroscopy
Streptomycin is a registered plant bactericide used for the control
of commercially important bacterial plant pathogens. It is usually
marketed as the sulfate, nitrate, or hydrochloride. The structure of
di-base tris-sulfate is:
H+
NH
H2N—C HN
<:HO
HN-CH3 OH
Streptomycin is a strongly basic compound with the empirical
formula C2,H gN70 „; molecular weight 581.6; it is triacidic and forms
salts with acids (as above where 2 molecules of the base combine with
3 molecules of sulfuric acid); it is not affected seriously by exposure
to light and air, but is hygroscopic and quite readily deliquesces;
its solutions are reasonably stable over the pH range 3 to 7; it is
stable when dry.
Streptomycin sulfate molecular formula: (C H N 0 ) 3H SO
(molecular weight: 1457.44) is a white or practically white powder; it
is odorless or has a very faint odor; it is hygroscopic, but stable
toward air and light; it is very slightly soluble in alcohol and
-------
2 Streptomycin EPA-1
practically insoluble in chloroform, but is freely soluble in water; its
solutions are acid to nearly neutral litmus.
Other names: Agrimycin, Agri-Strep, streptomycins (France), strepto-
mycin sulfate., streptomycin nitrate, streptomycin hydro-
chloride
Principle_of the Method;
Streptomycin compounds are subjected to an aqueous alkaline hydrolysis
to form maltol which is determined by UV at 324 nm in the aqueous alkaline
solution. Alternatively, the aqueous alkaline maltol solution can be
neutralized with acid, treated with ferric chloride to produce a purple-
red color, and determined by reading in the visible range at 530 nm.
Reagents:
1. Streptomycin (base or salt) standard of known % purity
2. Sodium hydroxide, IN solution
3. Hydrochloric acid, 1.2N solution
4. Hydrochloric acid, 0.1N solution
5. Ferric chloride, 10% solution
6. Ferric chloride, 0.25% solution - prepare fresh daily by
pipetting 2.5 ml 10% ferric chloride solution and 10 ml 0.1N
hydrochloric acid solution into a 100 ml volumetric flask and
make to volume with water.
Equipment;
1. Ultraviolet-visible spectrophotometer, double beam ratio
recording with matched 1 cm silica cells
2. Boiling water bath
3. Ice water bath
4. Usual laboratory glassware
-------
3 Streptomycin EPA-1
Procedure:
Preparation of Standard;
Weigh 0.12 gram streptomycin base or 0.15 gram streptomycin
sulfate into a 250 ml volumetric flask; dissolve in and make to
volume with water, and mix thoroughly. This solution must be
stored in a refrigerator and should be made fresh at least every
2 weeks, (cone O.A8 mg streptomycin base or 0.6 mg streptomycin
sulfate/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.12 gram strepto-
mycin base or 0.15 gram streptomycin sulfate into a 250 ml volu-
metric flask, dissolve in, make to volume with water, and mix
thoroughly, (cone 0.48 mg streptomycin base or 0.6 mg strepto-
mycin sulfate/ml)
Determination;
Pipette 10 ml of standard solution into a 25 ml volumetric
flask, 10 ml of sample solution into a second 25 ml volumetric
flask, and 10 ml water (for blank) into a third 25 ml volumetric
flask. Add, by pipette, 2 ml IN sodium hydroxide solution to each
of the 3 flasks and heat in a boiling water bath for 10 minutes.
Cool in an ice water bath for three minutes.
A determination in the ultraviolet region can be made at this
point by making each of the 3 flasks to volume with water, mixing
well, and diluting a 10 ml aliquot of each to 50 ml with water.
Standard and sample solutions are scanned from 360 nm to 260 run
using the blank solution as reference. Measure the analytical
peak at 324 nm.
For a colorimetric determination in the visible region, a
purple-red color is developed as follows: to each of the 3 flasks,
add 2 ml 1.2N hydrochloric acid to neutralize the sodium hydroxide,
-------
A Streptomycin EPA-1
add 5 ml 0.25% ferric chloride solution, make to volume with
water, and mix thoroughly. Scan the standard and sample solutions
from 650 nm to 450 nm using the blank solution as reference.
Measure the analytical peak at 530 nm.
Calculations:
From the absorbances and concentrations of standard and
sample, calculate the percent streptomycin base or streptomycin
sulfate as follows:
„ (abs. sample)(cone. standard)(% purity standard)
(abs. standard)(cone, sample)
% streptomycin sulfate * 1.253 x % streptomycin
% streptomycin = 0.7978 x % streptomycin sulfate
-------
December 1975
Strychnine EPA-1
Determination of Strychnine
in Poisoned Baits (Picric acid precipitation)
Strychnine is a registered rodenticide having the chemical
structure:
Molecular formula: C21H22N_02
Molecular weight: 334.4
Melting point: 268 to 290°C (depending on the speed of heating)
with decomposition; b.p. 270*C at 5 mm
Physical state, color, and odor: hard white crystals or powder, very
bitter taste; very poisonous!
Solubility: practically insoluble in water, alcohol, ether; slightly
soluble in benzene, chloroform
Stability: forms salts with acids; ppt by alkaloid precipitants
(e.g., picric acid as in this method)
Other names: Kwik-kil, Mouse-tox, Ro-Dec
-------
2 Strychnine EPA-1
'%
Strychnine generally Is used as the sulfate; poison baits usually
are colored grain containing 0.5 to 1Z strychnine sulfate.
Strychnine Sulfate is a white crystalline powder containing 5
moles of water of crystallization lost at 110°C; moderately soluble
in water and alcohol, insoluble in ether; mol. formula: (C..H..N.O ) _
H2SOA.5H20; mol. wt. 856.96; m.p. above 199°C.
Principle of the Method:
Strychnine is extracted from the poison bait formulations using
an ether-chloroform solvent mixture with some ammonium hydroxide
solution to convert salts to the free alkaloid. After lead acetate
and sodium oxalate treatments, the strychnine is precipitated with
picric acid and weighed as strychnine picrate.
Reagents:
g.-fhe.r
1. Ether-chloroform mixture (2 parts ethamrt + 1 part chloroform)
2. Ammonium hydroxide,, 10% solution
3. Corn syrup (such as white Karo)
4. Ethyl ether
5. Hydrochloric acid, 0.5% solution
6. Acetic acid
7. Neutral lead acetate, 10% aqueous solution
8. Sodium oxalate, 3% aqueous solution
9. Picric acid, saturated aqueous solution (1 g/100 ml)
All chemicals and solvents, ACS or reagent grade
-------
3 Strychnine EPA-1
Equipment:
1. Usual laboratory glassware
2. Filter paper (Whatman No. 1 and No. 30 or equivalent)
3. Gooch crucible, prepared with filter pad, dried, and weighed
Procedure;
Weigh a portion of finely ground sample equivalent to about 0.1
gram strychnine or 0.13 gram of strychnine sulfate into a 300 ml Erlen-
meyer flask. Add (conveniently at 3:00 p.m.) 150 ml of (2+1) ether-
chloroform mixture and stopper tightly. Allow to stand 30 minutes with
occasional agitation. Add 25 ml of 10% ammonium hydroxide solution,
shake one hour, and allow to stand overnight.
In the morning, shake for 15 minutes, add about 5 ml corn syrup
(such as white Karo) to clarify the solution, shake again for 15
minutes, and allow to settle. Four off 100 ml of the solvent layer
and transfer to a 250 ml separatory funnel. Add enough ether (approx.
50 ml) to cause the solvent layer to rise to the top in the subsequent
extractions. Extract with 0.5% hydrochloric acid, using a 50 ml portion
for the first extraction and a 25 ml portion for each of six additional
extractions. Collect the extracts in a 400 ml beaker. (A milky emulsion
will be formed on shaking, but this should be entirely drained off each
time.)
Evaporate the combined extracts to 50 ml, cool, and make alkaline
with ammonium hydroxide, avoiding an excess. Make slightly acid with
acetic acid and warm gently for a few minutes until a flocculation of
the suspended matter takes place. Cool, add 2 ml of 10% neutral lead
-------
4 Strychnine EPA-1
acetate solution, transfer to a 100 ml volumetric flask, make to volume,
and shake thoroughly. Filter through dry paper (Whatman #1 or equiv-
alent) into a dry 100 ml glass-stoppered graduated cylinder without
washing and note the volume obtained. Add 3.0 ml of 3% sodium oxalate
solution, shake thoroughly, and allow to stand for 15 minutes. Again
filter through a dry paper (Whatman No. 30 or equivalent) into a dry
100 ml glass-stoppered cylinder without washing and note the volume
obtained.
Transfer to a 250 ml beaker, evaporate to 70 ml, and cool. Add
25 ml of a recently filtered saturated picric acid solution and allow
to stand for 3 hours with occasional stirring during the first half
hour. Filter on a tared Gooch crucible and wash with 50-80 ml cold
water. Dry at 105°C and weigh.
Calculations:
? ci-r-^hn^o . (grams strychnine picrate) (0.5934) (100)
i strycnnine (go gample)(100/i50)(X/100)(Y/X + 3)
where: 0.5934 » factor for strychnine picrate to strychnine
X « ml collected from first filtration
(after lead acetate addition)
Y - ml collected from 2nd filtration
(after sodium oxalate addition)
% strychnine sulfate - % strychnine X 1.281
-------
December 1975
Strychnine EPA-2
Determination of Strychnine in Commercial
Bait Formulations by Ultraviolet Spectroscopy
Strychnine is a registered rodenticide having the chemical
structure:
Molecular formula: C2iH22N2°2
Molecular weight: 334.4
Melting point: 268 to 290°C (depending on the speed of heating)
with decomposition; b.p. 270°C at 5 mm
Physical state, color, and odor: hard white crystals or powder, very
bitter taste; very poisonous!
Solubility: practically insoluble in water, alcohol, ether; slightly
soluble in benzene, chloroform
Stability: forms salts with acids; ppt by alkaloid precipitants
(e.g., picric acid as in this method)
Other names: Kwik-kil, Mouse-tox, Ro-Dec
-------
2 Strychnine EPA-2
Principle of the Method;
Strychnine is extracted from the sample with a 0.5% sulfuric acid
solution. The extract is cleaned up and the strychnine determined by
the difference in absorbance at 254 and 287 ran using a concentration
of 10-20 ug/ml.
This method is not suitable for commercial strychnine sulfate
formulations. The rodenticide seems to be complexed or associated
with the carrier in these products, and the strychnine sulfate is not
quantitatively extracted by the sulfuric acid solution. (Use EPA-1
for the sulfate)
Reagents;
1. Strychnine standard of known % purity
2. Sulfuric acid solution, 0.5% V/V solution
3. Concentrated ammonium hydroxide
4. Chloroform, ACS
5. Ethyl ether, ACS
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm absorption cells
2. Mechanical shaker
3. Ultrasonic cleaner (useful for dissolving standard but not
essential)
4. Steam bath
5. Hot plate
6. Usual laboratory glassware
-------
3 Strychnine EPA-2
Procedure:
Preparation of Standard;
Weigh 0.1 gram strychnine standard into a 100 ml volumetric
flask, add about 90 ml 0.5% sulfuric acid solution, stopper
tightly, and shake to dissolve the strychnine. (Dissolution of
the strychnine may be hastened by placing the volumetric flask in
an ultrasonic bath for a few minutes.) Make to volume and mix
thoroughly. Pipette 15 ml into a second 100 ml volumetric flask,
make to volume vith the 0.5Z sulfuric acid solution, and mix
thoroughly.
Prepare three dilutions by pipetting 5, 10, and 15 ml into
separate 100 ml volumetric flasks and making each to volume with
0.52 sulfuric acid. Mix each flask thoroughly, (final cones
7.5, 15.0, and 22.5 ;ig/ml)
(If a direct standard - sample comparison is to be made,
use 15 ^ig/ml cone for the standard.)
Preparation of Sample;
Uniformly coated bait materials may be used directly but
non-uniform materials should be ground to a fine powder.
Weigh a portion of sample equivalent to 0.025 gram of strychnine
into a 250 ml glass-stoppered or screw-cap flask, add by pipette
100 ml 0.5% sulfuric acid solution, and shake on a mechanical shaker
for 6 hours. Let sample stand overnight. Shake an additional half-
hour the next day, allow to settle, and filter. Transfer a 25 ml
aliquot into a 100 ml volumetric flask, and make to volume with 0.5%
sulfuric acid solution. Mix thoroughly and pipette 25 ml into a
125 ml separatory funnel.
Add 2 ml concentrated ammonium hydroxide to the separatory
funnel and shake. The solution should be basic; if not, add more
ammonium hydroxide. Extract with four 25 ml portions of chloroform,
-------
A Strychnine EPA-2
draining each extract through plug of cotton (prevashed with chloro-
form) into AGO ml beaker. Transfer all emulsions which form during
the extraction onto the cotton. Extract the solution once more
with 50 ml chloroform and drain through the cotton. Wash the
cotton with 15 ml chloroform and squeeze out the excess.
Add three glass beads to the beaker and evaporate the chloro-
form extract to dryness on a steam bath. Heat until all the
chloroform vapor is dissipated. Cool, dry the exterior of the
beaker, and add AO ml 0.5% sulfuric acid solution. Weigh the
beaker (with a stirring rod) to two decimal places. Heat on steam
bath 20-30 min, bringing liquid into contact with the residue on the
side of the beaker, and re-weigh. Add an amount of water to the
beaker equal to the weight of that evaporated. (Note: It is
desirable to keep the environment of the sample a lose to, or
identical with, that of the reference standard In absorption spec-
troscopy. For this reason the evaporated water Is added twice in
handling the sample. The acid concentrations in the standard and
sample are, for all practical purposes, the same. However, no
appreciable analytical error would be expected if the acid concen-
tration in the sample was significantly weaker than that of the
standard.)
Transfer the solution to 250 ml separatory funnel, washing the
beaker with two 10 ml portions of 0.5% sulfuric acid solution and
adding the liquid to the separatory funnel. Add 50 ml ethyl ether
to the separatory funnel and shake for 1 min. (The extraction with
ether removes fatty acids or oils which may be present in the
strychnine sample.) Drain the aqueous layer into a 250 ml beaker.
Wash the ethyl ether layer with two 5 ml portions of 0.5% sulfuric
acid solution and add to the beaker. Add three glass beads to
beaker and weigh beaker to two decimal places. Heat the liquid to
boiling on a hot plate to remove dissolved ether and evaporate to
ca 40 ml. Cool to room temperature, dry exterior of beaker, and
weigh. Return an amount of water to the beaker equal to that
evaporated (see note above).
-------
5 Strychnine EPA-2
Transfer the solution to a 100 ml volumetric flask and make to
volume with 0.52 sulfuric acid solution; mix thoroughly, (final
cone 15.6 ug strychnine /ml)
UV Determination;
With the UV spectrophotoraeter at the optimum quantitative
settings for the particular instrument being used, balance the
pen for 0 and 100% transmission at 254 nm with 0.5% sulfuric acid
in each cell. Scan the standard and sample solutions from 350 nm
to 225 nm with 0.5% sulfuric acid solution in the reference cell.
Calculation :
Determine the difference in absorbance at 254 and 287 nm
(A - abs(254 nm) - abs(287 nm)) for standards and sample. Plot
an absorbance vs. concentration curve for the three standards
(Beer's law is obeyed), and calculate the percentage strychnine
in the sample from the standard curve as follows:
y „ (abs. sample )(conc. standard) (purity of standard) (100)
(abs. stand.) (cone, sample)
The percent strychnine may be determined using a direct standard -
sample comparison (without using a standard curve) as follows:
_spl)(conc. std in ug/ml) (% purity std)
-------
December 1975 Sulfur EPA-1
Determination of Free Sulfur In
Sulfur Formulations (CS Extraction)
Sulfur Is a registered fungicide and acaricide.
Molecular (atomic) formula: S
Molecular (atomic) weight: 32.06
Melting point: 115°C; b.p. 444.6°C
Physical state and color: yellow solid, melting at 115°C to a yellow
mobile liquid which darkens and becomes viscous about
1608C. It exists in two allotropic forms: rhombic,
m.p. 112.8°C, and monoclinic, m.p. 119'C.
Solubility: practically insoluble in water, slightly soluble in
ethanol and ether; the crystalline forms are soluble in
carbon disulfide whereas the amorphous forms are not.
Stability: compatible with most other pesticides, except petroleum
oils; slowly hydrolyzed by water (detectable when a
product of hydrolysis is removed, as in the tarnishing
of silver or its reaction with alkalis)
Other names: Brimstone; Flowers of sulfur (» sublimed sulfur); Flour
sulfur (- ground rock sulfur); precipitated sulfur
Reagents;
1. Carbon disulfide, ACS
Equipment:
1. Filtration apparatus
2. Exhaust hood
3. Steam bath
4. Drying oven (100-105'C)
5. Usual laboratory glassware
-------
Sulfur EPA-1
Procedure;
Weigh a portion of sample equivalent to about 0.1 gram of sulfur
and transfer to a funnel fitted with dry filter paper. Wash the sample
with small portions of dry carbon disulfide, catching the filtrate in a
dry weighed beaker. Continue washing until the sulfur is apparently all
extracted, (see note at end of procedure)
Evaporate the carbon disulfide in an exhaust hood either over a
steam bath or spontaneously at room temperature. (CAUTION - carbon
disulfide is extremely flammable!) When the carbon disulfide is com-
pletely evaporated, heat the beaker and residue for 15-20 minutes at
100-105°C and weigh. Subtract to determine the weight of elemental
sulfur.
Using the above weight, calculate the percent sulfur in the sample
as follows:
'/ if (wt. elemental sulfur) (100)
*9 O U JLI U t ._ i.-_ir- •--«. - j- r -M-. .- _ -.
(wt. sample)
Note;
A portion of the sulfur may be present as flowers of sulfur and is
not soluble in carbon disulfide. In such cases, the sulfur must be
determined by oxidation and precipitation as barium sulfate - see method
Sulfur EPA-2. The determined sulfur, calculated to elemental sulfur, is
added to the above result to obtain total free sulfur.
If there are any sulfates present in the sample, determine these on
a hydrochloric acid solution of the original sample and subtract from the
total sulfur determined on the carbon disulfide washed residue. The
difference, calculated to elemental sulfur, represents the sulfur from
the undissolved flowers of sulfur. This should be added to the carbon
disulfide soluble sulfur to give the total free sulfur in the sample.
-------
December 1975 Sulfur EPA-2
Determination of Sulfur by Oxidation
and Precipitation as Barium Sulfate
Sulfur is a registered fungicide and acaricide.
Molecular (atomic) formula: S
Molecular (atomic) weight: 32.06
Melting point: 115°C; b.p. 444.6°C
Physical state and color: yellow solid, melting at 115°C to a yellow
mobile liquid which darkens and becomes viscous about
160°C. It exists in two allotropic forms: rhombic,
m.p. 112.8"C, and monoclinic, m.p. 119°C.
Solubility: practically insoluble in water, slightly soluble in
ethanol and ether; the crystalline forms are soluble in
carbon disulfide whereas the amorphous forms are not.
Stability: compatible with most other pesticides, except petroleum
oils; slowly hydrolyzed by water (detectable when a
product of hydrolysis is removed, as in the tarnishing
of silver or its reaction with alkalis)
Other names: Brimstone; Flowers of sulfur (» sublimed sulfur); Flour
sulfur (= ground rock sulfur); precipitated sulfur
Reagents;
1. Fuming nitric acid (specific gravity 1.49-1.50)
2. Concentrated hydrochloric acid
3. 10% Barium chloride solution
-------
2 Sulfur EPA-2
Equipment:
1. 300 ml Erlenmeyer soil flask with an air condenser connected
by ground glass joints
2. Steam bath
3. Hot plate
4. Filtration apparatus
5. Platinum Gooch crucible, previously ignited and weighed
6. Muffle furnace
7. Usual laboratory glassware
Procedure:
Weigh a portion of sample equivalent to 0.025 to 0.035 gram sulfur
into a 300 ml Erlenmeyer soil flask fitted with an air condenser by
means of a ground glass joint. Add cautiously (through the condenser)
25 ml fuming nitric acid in small portions, taking about 15 minutes to
make the addition so that the reaction does not become violent. Let
stand for one-half hour, swirling gently from time to time to mix
thoroughly. Heat gently on a covered steam bath, and when the reaction
slows, heat in direct contact with steam for one hour.
Cool, wash down the inside of the condenser, and quantitatively
transfer the contents of the flask to a beaker. Evaporate to dryness,
add 3 ml hydrochloric acid, and again evaporate to dryness. Repeat the
addition of hydrochloric acid and the evaporation to dryness two more
times. Dissolve the residue in about 5 ml water and 5 ml hydrochloric
acid, quantitatively transfer to a 250 ml volumetric flask, make to
volume with water, and mix thoroughly.
-------
3 Sulfur EPA-2
Pipette a 50 ml aliquot into a 600 ml beaker, dilute to about 400 ml
with water, and add 10 ml hydrochloric acid. Heat nearly to boiling and
add slowly, dropwise with stirring, sufficient 10% barium chloride
solution to precipitate the sulfur as barium sulfate. Wash down the
sides of the beaker, and heat just under the boiling point for one hour.
Filter through a previously ignited and weighed Gooch crucible,
wash, dry, and ignite in a muffle furnace at 550-650°C. Weigh as
barium sulfate.
Calculate the percent sulfur in the sample as follows:
. „ . ,£ ^ (wt. of precit>itate)(100)
% Barium sulfate - - E ,;•,.. , l^\
(wt. sample)(50/250)
% Sulfur - (0.1374)(% barium sulfate)
-------
December 1975 Sulfur EPA-3
Determination of Sulfur in Dusting Mixtures
in the Presence of Acetone-Soluble Pesticides
Sulfur is a registered fungicide and acaricide.
Molecular (atomic) formula: S
Molecular (atomic) weight: 32.06
Melting point: 115°C; b.p. 444.6°C
Physical state and color: yellow solid, melting at 115°C to a yellow
mobile liquid which darkens and becomes viscous about
160°C. It exists in two allotropic forms: rhombic,
m.p. 112.8°C, and monoclinic, m.p. 119°C.
Solubility: practically insoluble in water, slightly soluble in
ethanol and ether; the crystalline forms are soluble in
carbon disulfide whereas the amorphous forms are not.
Stability: compatible with most other pesticides, except petroleum
oils; slowly hydrolyzed by water (detectable when a
product of hydrolysis is removed, as in the tarnishing
of silver or its reaction with alkalis)
Other names: Brimstone; Flowers of sulfur (» sublimed sulfur); Flour
sulfur (» ground rock sulfur); precipitated sulfur
Reagents;
1. Acetone, sulfur—saturated - prepare by adding an excess of
sulfur to acetone, warm gently to effect solution, then cool
to room temperature. Filter before using.
2. Carbon disulfide, ACS
-------
2 Sulfur EPA-3
Equipment;
1. 125 ml glass-stoppered flask, preferably with a pour-out lip
2. Filter paper equivalent to S&S No. 590 or Whatman No. 40
3. Short-stemmed funnel
4. Dry, weighed 150 ml beaker
Procedure;
Weigh a portion of sample equivalent to about 0.2-0.3 gram sulfur
into a glass-stoppered 125 rul Erlenmeyer flask (preferably with pour-
out lip), add 50 ml of the sulfur-saturated acetone, stoppered tightly,
and shake for several minute:s to dissolve all the acetone-soluble
pesticides and other acetone-soluble substances. Filter, transferring
the insoluble residue containing the sulfur to the paper with small
portions of sulfur-saturated acetone. Wash the residue several times
with small portions of the sulfur-saturated acetone to remove all traces
of acetone-soluble substances.
Allow the acetone to volatilize from the original flask and filter
paper, place a dry, weighed 150 ml beaker under the funnel, and wash the
flask and residue with carbon disulfide. Continue the washing of the
residue with carbon disulfida until all the sulfur has apparently been
removed. Evaporate the carbon disulfide gently on a steam bath. When
the odor of carbon disulfide is no longer present, dry in an oven at
105°C for 15 minutes. CAUTION - carbon disulfide is extremely flammable!
-------
3 Sulfur EPA-3
Cool, weigh, and calculate the percent carbon disulfide soluble
sulfur as follows:
% Sulfur residue)J100).
(wt . sample)
Note: The recovered sulfur should be free of plant extractives;
however, if it appears to contain small quantities, they may
be removed as follows:
Add 25 ml of the sulfur-saturated acetone and with the
aid of a rod flattened on one end, disintegrate the residue
in such a manner that acetone comes in contact with all the
sulfur crystals. Filter the dissolved plant extractives
through a weighed Gooch crucible that has been fitted with a
disk of filter paper. Rinse the sulfur from the beaker into
the paper and wash under suction with the sulfur-saturated
acetone. Allow the acetone to evaporate under suction for about
10 minutes; then dry the crucible in an oven at 105°C for 15
minutes. Cool, weigh, and re-calculate the percent sulfur as
above.
Should the sample contain flowers of sulfur or be below the
declared percentage, determine sulfur by EPA-2.
-------
December 1975 Sulfur Dioxide EPA-1
Determination of Sulfur Dioxide
in Fumigants by lodometry
Sulfur dioxide is a registered fumigant, having the chemical
structure:
0=S->0
Molecular formula: SO.
Molecular weight: 64.07
Boiling point: -10°C
Physical state, color, and odor: colorless gas with a strong suffocating
odor characteristic of burning sulfur; under pressure
condenses readily to a colorless liquid
Solubility: soluble in water, alcohol, ether, chloroform; forms sulfurous
acid, H SO with water
Stability: nonflammable; an outstanding oxidizing and reducing agent;
CAUTION - extremely irritating to eyes and respiratory tract
Other names: sulfurous acid anhydride, sulfurous oxide
Reagents:
1. Iodine solution, 0.1N standardized solution
2. Sodium thiosulfate solution, 0.1N standardized solution
3. Acetic acid, ACS
Equipment;
1. Titration apparatus
2. Usual laboratory glassware
-------
2 Sulfur Dioxide EPA-1
Principle _gf the Method;
Since sulfur dioxide is volatile, the product container should not
be opened until just before the sample portion is to be removed. Loss
of sulfur dioxide is minimized by weighing the sample, by difference,
directly in a known amount of acidified iodine solution. The excess is
titrated and the sulfur dioxide calculated from the iodine solution
used.
Procedure;
Pipette 50 ml 0.1N iodine solution into a 125 ml glass-stoppered
flask, add 5 ml acetic acid, stopper, and weigh accurately.
Transfer a portion of sample equivalent to 0.1 gram sulfur dioxide
into the flask with swirling, restopper, weigh, and obtain the sample
weight by difference.
Titrate the excess iodine solution with 0.1N sodium thiosulfate
solution. Starch indicator is usually not necessary, but may be used
close to the end of the titration.
Calculation:
Calculate the sulfur dioxide as follows:
% m [(ml I2)(NI2) - (ml Na2S203)(N Na^O^] (0.03203) (100)
(wt. sample in grams)
If an identical 50 ml portion of the 0.1N iodine solution is
titrated (without sample), then calculate the sulfur dioxide as
follows:
„ (ml difference Na,S90,)(N Na,S,0,)(0.03203)(100)
M £. jL jt £. JL j
(wt. sample in grams)
-------
-------
December 1975
Terbutol EPA-1
(Tentative)
Determination of Terbutol
by Infrared Spectroscopy
Terbutol is the common name (WSSA) for 2,6-di-tert-butyl-p-tolyl
methylcarbamate, a registered herbicide having the chemical structure:
0 H
C—N—CH3
Molecular formula: C ?H NO-
Molecular weight: 277.4
Melting point: 200 to 201°C; the technical product is 95% and has
a mp of 185 to 190°C
Physical state, color, and odor: white, odorless, crystalline solid
Solubility: 7 ppm in water at 25°C; insoluble in hexane and kerosene;
slightly soluble in benzene and toluene; soluble in acetone
and ethanol
Stability: decomposes at melting point; nonflammable; compatible with
hard water, other pesticides, and fertilizer; non-corrosive;
stable on storage
Other names: Azak (Hercules, Inc.), Hercules 9573, Terbucarb
-------
2 Terbutol EPA-1
(Tentative)
Reagents;
1. Terbutol standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm NaCl or KBr cells
2. Mechanical shaker
3. Usual laboratory glassware
Procedure;
Preparation of Standard;
Weigh 0.08 gram terbutol standard into a small glass-stoppered
flask or screw-cap bottle, add 10 ml chloroform by pipette, close
tightly, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 8 mg/ml)
Preparation of Sample:
Weigh an amount of sample equivalent to 0.8 gram terbutol into
a glass-stoppered flask or screw-cap tube. Add 100 ml chloroform
by pipette and 1-2 grams anhydrous sodium sulfate. Close tightly
and shake for one hour. Allow to settle; centrifuge or filter if
necessary, taking precautions to prevent evaporation, (final cone
8 mg terbutol/ml)
-------
Terbutol EPA-1
(Tentative)
De termina tion;
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1925 cm to
1580 cm"1 (5.2 p to 6.3 ;i).
Determine the absorbance of standard and sample using the peak
L754 cm"1 (
(5.45 ^ to 5.9
Calculation:
at 1754 cm" (5.7 ;i) and a baseline from 1835 cm" to 1695 cm~
From the above absorbances and using the standard and sample
solution concentrations, calculate the percent terbutol as follows
„ (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Beer's law is obeyed over the range 1-14 mg/ml.
Method submitted by Dean Hill, EPA Region IX, San Francisco, Calif.
-------
December 1975
Terbutol EPA-2
(Tentative)
Determination of Terbutol by
Gas-Liquid Chromatography
(FID - Internal Standard)
Terbutol is the common name (WSSA) for 2,6-di-tert-butyl-p-tolyl
methylcarbamate, a registered herbicide having the chemical structure:
CH3— C— CH3
Molecular formula: C _H NO
Molecular weight: 277.4
Melting point: 200 to 201°C; the technical product is 95% and has
a mp of 185 to 190°C
Physical state, color, and odor: white, odorless, crystalline solid
Solubility: 7 ppm in water at 25°C; insoluble in hexane and kerosene;
slightly soluble in benzene and toluene; soluble in acetone
and ethanol
Stability: decomposes at melting point; nonflammable; compatible with
hard water, other pesticides, and fertilizer; non-corrosive;
stable on storage
Other names: Azak (Hercules, Inc.), Hercules 9573, Terbucarb
-------
2 Terbutol EPA-2
(Tentative)
Reagents:
1. Terbutol standard of known % purity
2. Diazinon standard of known % purity
3. Acetone, pesticide or spectre grade
A. Internal Standard solution - weigh 0.2 gram diazinon into a
100 ml volumetric flask; dissolve in and make to volume with
acetone, (cone 2 mg diazinon/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm I.D. glass column packed with 5% SE-30 on
80/100 Chromosorb W HP (or equivalent column)
3. Precision liquid syringe: 5 or 10 pi
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 165°C
Injection temperature: 215°C
Detector temperature: 215°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi (adjusted for specific GC)
Hydrogen pressure: 20 psi (adjusted for specific GC)
Air pressure: 30 psi (adjusted for specific GC)
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and
reproducibility.
-------
Terbutol EPA-2
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.05 gram terbutol standard into a small glass-stoppered
flask or screw-cap bottle, add by pipette 25 ml of the internal
standard solution, and shake to dissolve, (final cone 2 mg
terbutol and 2 mg diazinon/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.05 gram terbutol
into a small glass-stoppered flask or screw-cap bottle; add by
pipette 25 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the terbutol. For
coarse or granular materials, shake mechanically for 30 minutes
or shake by hand intermittently for one hour (final cone 2 mg
terbutol and 2 mg diazinon/ul)
Determination:
Inject 1-2 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is diazinon, then terbutol.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of terbutol and diazinon from
both the standard-internal standard solution and the sample-
internal standard solution.
-------
Terbutol EPA-2
(Tentative)
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
= (wt. diazinon)(% purity diazinon)(pk. ht. or area terbutol)
(wt. terbutol)(% purity terbutol)(pk. ht. or area diazinon)
Determine the percent terbutol for each injection of the sample-
internal standard solution as follows and calculate the average:
„ = (wt. diazinon)(% purity diazinon)(pk. ht. or area terbutol)ft6fr);
(wt. sample) (pk. ht. or area diazinon) (RF) (M-~l)~
This method was submitted by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond, Virginia
23219.
Note; This method has been designated as tentative since it is a
Va. Exp. method and because some of the data has been suggested
by EPA's Beltsville Chemistry Lab. Any comments, criticisms,
suggestions, data, etc. concerning this method will be appreciated.
-------
September 1975 Thiram EPA-1
Determination of Thiram
by Ultraviolet Spectroscopy
Thiram is the official common name for tetramethylthiuram
disulfide, a registered fungicide having the chemical structure:
•C—N
XCH3
Molecular formula: C,H 2N S
Molecular weight: 240.44
Melting point: 155 to 156CC
Physical state and color: colorless crystals
Solubility: about 30 ppm in water at RT; slightly soluble in ethanol,
ether, carbon disulfide; soluble in acetone, chloroform
Stability: stable in storage; in the form of a fine dust it gives
explosive mixtures with air.
Other names: Arasan (DuPont), Nomersan (Plant Protection Ltd.),
Pomarsol (I. G. Farb.), Tersan, Thylate Spotrete,
Thimar, Mercuram, Tuads, Vancide, Hexathir, Fermide,
Bis(dimethylthiocarbamoyl)disulphide, TMTD
Reagents;
1. Thiram standard of known % purity
2. Chloroform, pesticide or spectro grade
-------
2 Thiram EPA-1
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm silica cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure;
Preparation of Standard:
Weigh 0.1 gram thiram standard into a 100 ml volumetric
flask, add 100 ml chloroform by pipette, and mix thoroughly.
Pipette 10 ml into a second 100 ml volumetric flask, make to
volume with chloroform, and mix thoroughly. Pipette 10 ml into
a third 100 ml volumetric flask, make to volume with chloroform,
and mix thoroughly, (final cone 10 jig/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.1 gram thiram
into a 250 ml glass-stoppered or screw-cap flask, add 100 ml
chloroform by pipette, and shake on a mechanical shaker for 30
minutes. Allow to settle; centrifuge or filter if necessary,
taking precautions to prevent evaporation. Pipette 10 ml into
a 100 ml volumetric flask, make to volume with chloroform, and
mix thoroughly. Pipette 10 ml of this solution into another
100 ml volumetric flask, make to volume with chloroform, and mix
thoroughly, (final cone 10 ^ig thiram/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen at 0 and 100% transmission at 280 nm with
-------
Thiram EPA-1
chloroform in each cell. Scan both the standard and sample from
350 tun to 250 nm with chloroform in the reference cell.
Measure the absorbance of standard and sample at 280 nm.
Calculation;
From the above abaorbances and using the standard and sample
concentrations, calculate the percent thiram as follows:
. m (abs. sample)(cone, std in jug/ml)(2 purity std)
(abs. std)(cone, sample in jug/ml)
-------
August 1975 Thiram EPA-2
Determination of Thiram
by Infrared Spectroscopy
Thiram is the official common name for tetramethylthiuram
disulfide, a registered fungicide having the chemical structure:
rw S S
CH3\ II II
\ - C - S - S — C
Molecular formula: C,H, -N.S.
6 12 2 4
Molecular weight: 240.44
Melting point: 155 to 156°C
Physical state and color: colorless crystals
Solubility: about 30 ppm in water at RT; slightly soluble in ethanol,
ether, carbon disulfide; soluble in acetone, chloroform
Stability: stable in storage; in the form of a fine dust it gives
explosive mixtures with air.
Other names: Arasan (DuPont) , Nomersan (Plant Protection Ltd.),
Pomarsol (I. G. Farb.), Tersan, Thylate, Spotrete,
Thimar, Mercuram, Tuads, Vancide, Hexathir, Fermide,
Bis(dimethylthiocarbamoyl)disulphide, TMTD
Reagents;
1. Thiram standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
2 Thiram EPA-2
Equipment;
1. Infrared spectrophotometer, double beam ratio recording with
matched 0.2 mm Nad or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.065 gram thiram standard into a small glass-
stoppered flask or screw-cap bottle, add 10 ml chloroform by
pipette, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 6.5 mg/ml)
Preparation of Sample;
For dusts, granules, and wettable powder, weigh a portion of
sample equivalent to 0.325 gram thiram into a glass-stoppered
flask or screw-cap bottle. Add 50 ml chloroform by pipette and
1-2 grams anhydrous sodium sulfate. Close tightly and shake for
one hour. Allow to settle; centrifuge or filter if necessary,
taking precaution to prevent evaporation, (final cone 6.5 mg
thiram/ml) For very low percent formulations requiring larger
samples,use more solvent and evaporate an aliquot to a smaller
volume to give a concentration close to 6.5 mg thiram/ml.
-------
3 Thiram EPA-2
For water suspensions a tentative procedure is as follows:
weigh a portion of sample equivalent to 0.325 gram thiram into
a glass-stoppered flask or screw-cap bottle. Add 50 ml chloro-
form by pipette and sufficient anhydrous sodium sulfate to
absorb the water and dry and clarify the chloroform solution;
shake thoroughly, (final cone 6.5 mg thiram/ml)
Determination:
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular 1R instrument
being used, scan both standard and sample from 1430 cm to
1300 cm"1 (7 p. to 7.7 ;u).
Determine the absorbance of standard and sample using the
peak at 1380 cm"1 (7.25 p) and baseline from 1400 cm"1 to 1350 cm~]
(7.14 ji to 7.41 ju).
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent thiram as follows:
7 - (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std) (cone, sample in tng/ml)
(A concentration of 1 mg thiram/ml chloroform gives an
absorbance of approx. 0.046 in a .2 mm cell.)
Method submitted by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services.
(The procedure for water suspensions has successfully been used by
EPA's Beltsville Chemistry Lab.)
-------
January 1976
Trlchlorocarbanillde EPA-1
Determination of Trichlorocarbanilide
in Detergents by Ultraviolet Spectroscopy
Trichlorocarbanilide is 3,4,4'-trichlorocarbanilide, a registered
bacteriostat and fungistat having the chemical structure:
Molecular formula: C _H Cl N 0
Molecular weight: 315.6
Melting point: 250°C (minimum)
Physical state, color, and odor: fine white powder; no odor or a
slight characteristic odor
Solubility: slightly soluble in dioxane, propylene glycol; soluble in
acetone, methyl isobutyl ketone, dimethyl formamide, alcohol
Stability: stable to light and heat; does not discolor by reaction with
other materials
Other names: TCC
Reagents;
1. 3,4,4'-trichlorocarbanilide standard of known % purity
2. Ethanol, pesticide or spectro grade
-------
2 Trichlorocarbanilide EPA-1
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording with
matched 1 cm cells
2. Steam bath
3. Filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard;
Weigh 0.07 gram trichlorocarbanilide standard into a 100 ml
volumetric flask? dissolve in (warming if necessary) and make to
volume with ethanol; mix thoroughly. Pipette 10 ml into a second
100 ml volumetric flask, make to volume with ethanol, and mix
thoroughly. Pipette 5 ml into a third 100 ml volumetric flask,
make to volume with ethanol, and again mix thoroughly. (final
cone 3.5 jig/ml)
Preparation of SampJLe:
Weigh a portion of sample equivalent to 0.0035 gram trichloro-
carbanilide (0.7 gram for a 0.5% formulation) into a 100 ml beaker,
add 40 ml ethanol, cover with a watch glass, and warm on a steam
bath. Filter, collecting the filtrate in a 100 ml volumetric flask.
Wash the residue in the beaker by adding another 40 ml ethanol,
warming, filtering, and adding the filtrate to the volumetric flask.
Transfer the residue from the beaker into the filter and wash with
warm alcohol. Cool the extracts and washing in the volumetric flask,
make to volume with ethanol, and mix thoroughly. Pipette 5 ml into a
50 ml volumetric, make to volume with alcohol, and mix thoroughly.
(final cone 3.5 jig trichlorocarbanilide/ml)
-------
Trichlorocarbanilide EPA-1
UV Pet ermination:
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 265 n» with ethanol
in each cell. Scan both the standard and sample from 300 nm to
210 nm with distilled water in the reference cell. Measure the
absorbance of both standard and sample at 265 nm. (A slight shift
to a lower wavelength may occur if moderate interference is present.)
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent trichlorocarbanilide as follows;
v K jabs- sample) (cone, std in jug/ml);_(_% purity std)
(abs. std) (cone, sample in/ig/ral)
-------
November 1975
Trifluralin EPA-1
Determination of Trifluralin by
Gas-Liquid Chromatography
(FID - Internal Standard)
Trifluralin is the accepted common name for 0',a?,a-trifluoro-2,6-
dinitro - N,N-dipropyl-p-toluidine, a registered herbicide having the
chemical structure:
CH3—CH2—CH2—N—CH2—CH2 —CH3
Molecular formula: C.,H,,F,N,0.
U lo 3 J <*
Molecular weight: 335.3
Melting point: 48.5 to 49.0°C (tech. product is at least 95% pure
and has a mp greater than 42°C)
Physical state, color, and odor: orange crystalline solid; no
appreciable odor
Solubility: less than 1 ppm in water at 27°C; 7% in ethanol, 40% in
acetone, 58% in xylene; soluble in other organic solvents
Stability: stable but susceptible to photochemical decomposition
Other names: Treflan (Eli Lilly), Trefanocide, Treficon, Triflurex,
Su Seguro Carpidor
-------
2 Trlfluralin EPA-1
Reagents;
1. Trifluralin standard of known % purity
2. Diisobutylphthalate
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.3 gram of diisobutyl-
phthalate into a 25 ml volumetric flask, dissolve in, and
make to volume with acetone, (cone 12 mg diisobutylphthalate/ml)
Equipment t
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 4 mm glass column packed with 5% SP-2401 on
80/100 mesh Supelcoport AW DMCS (or equivalent
column)
3. Precision liquid syringe: 5 or 10 jil
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FTP;
Column temperature: 200°C
Injection temperature: 210*C
Detector temperature: 275°C
Carrier gas: Nitrogen
Carrier gas pressure: (not stated in method)
Hydrogen pressure: 30 psi
Air pressure: 30 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
3 Trifluralin EPA-i
Procedure;
Preparation of Standard:
Weigh 0.13 gram trifluralin standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 10 ml of the
internal standard solution and shake to dissolve, (final cone
13 mg trifluralin and 12 rag diisobutylphthalate/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.13 gram trifluralin
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 10 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the trifluralin. For
coarse or granular materials, shake mechanically for 30 minutes or
shake by hand intermittently for one hour, (final cone 13 mg
trifluralin and 12 mg diisobutylphthalate/ml)
Determination:
Inject 2-3 pi of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is trifluralin, then diiso-
butylphthalate.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of trifluralin and diiso-
butylphthalate from both the standard-internal standard solution
and the sample-internal standard solution.
-------
Trifluralin EPA-1
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
I.S. « internal standard • diisobutylphthalate
Rp m (wt. I.S.)(% purity I.S.)(pk. ht. or area trifluralin)
(wt. trifluralin)(% purity trifluralin)(pk. ht. or area I.S.)
Determine the percent trifluralin for each injection of the
sample-internal standard solution as follows and calculate the
average :
. I.S.)(% purity I.S.)(pk. ht. or area trifluralin)
(wt. sample) (pk. ht. or area I.S.)(RF)
Method submitted by Division of Regulatory Services, Kentucky Agricul-
tural Experiment Station, University of Kentucky, Lexington, Kentucky 40506.
-------
August 1975
Trifluralin EPA-2
Determination of Trifluralin
by Infrared Spectroscopy
Trifluralin is the accepted common name for a,<*,o'-trifluoro-
2,6-dinitro-N,N-dipropyl-p-toluidine, a registered herbicide
having the chemical structure:
CH3— CH2—CH2— N—CH2
-CH-
Molecular formula: Ci3HiAF3N304
Molecular weight: 335.3
Melting point: 48.5 to 49.0°C (tech. product is at least 95% pure
and has a mp greater than 42°C)
Physical state, color, and odor: orange crystalline solid; no
appreciable odor
Solubility: less than 1 ppm in water at 27°C; 7% in ethanol, 40% in
acetone, 58% in xylene; soluble in other organic solvents
Stability: stable but susceptible to photochemical decomposition
Other names: Treflan (Eli Lilly), Trefanocide, Treficon, Triflurex,
Su Seguro Carpidor
-------
2 Trifluralin EPA-2
Reagents;
1. Trifluralin standard of known % purity
2. Acetone, pesticide or spectro grade
3. Carbon disulfide, pesticide or spectro grade
4. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.2 mm NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.08 gram trifluralin into a small glass-stoppered
flask or screw-cap bottle, add 20 ml carbon disulfide by
pipette, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (final cone 4 rag/ml)
-------
3 Trifluralin EPA-2
Preparation of Sample;
For emulsifiable concentrates, weigh a portion of sample
equivalent to 0.04 gram trifluralin into a 10 ml volumetric
flask, make to volume with carbon disulfide, and mix well.
Add a small amount of anhydrous sodium sulfate to insure
dryness. (final cone 4 mg trif luralin/ml)
For granular formulations, weigh a portion of sample
equivalent to 0.08 gram trifluralin into a glass-stoppered
flask or screw-cap bottle. Add 50 ml acetone by pipette and
1-2 grams anhydrous sulfate. Close tightly and shake for one
hour. Allow to settle; centrifuge or filter if necessary,
taking precaution to prevent evaporation. Evaporate a 25 ml
aliquot to dryness on a water bath using a gentle stream of
dry air; evaporate the last one or two ml with air only.
Dissolve in about 4-5 ml carbon disulfide, transfer to a 10 ml
volumetric flask, and make to volume with carbon disulfide.
Add a small amount of anhydrous sodium sulfate to insure dryness.
(final cone A mg trif luralin/ml)
De t ermina t ion :
With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings for the particular IR
instrument being used, scan the standard and sample from 1390 cm"
to 1212 cm"1 (7.2 i to 8.25 i) .
Determine the absorbance of standard and sample using the
at 1300 cm"
(7.6 ^i to 7.91 |i).
peak at 1300 cm" (7.69 ji) and baseline from 1315 cm"1 to 1264 cm"1
-------
4 Trifluralin EPA-2
Calculation;
From the above absorbances and using the standard and
sample concentrations, calculate the percent trifluralin as
follows:
„ _ (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg trifluralin/ml carbon disulfide
gives an absorbance of approx. 0.079 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services.
-------
August 1975 Vernolate EPA-1
Determination of Vernolate
by Infrared Spectroscopy
Vernolate is the common name for S-propyl dipropylthiocarbamate,
a registered herbicide having the chemical structure:
| /CH2—CH2—CH3
CH3 —CH2—CH2—S—C N<^
XCH2 —CH2—CH3
Molecular formula: C H NOS
Molecular weight: 203.4
Boiling point: 140°C at 20 mm Hg, 150°C at 30 mm Hg
Physical state, color, and odor: clear liquid with an aromatic odor
Solubility: about 100 ppm in water at 20-21°C; miscible with
common organic solvents
Stability: stable; non-corrosive
Other names: Vernam (Stauffer), R-1607, S-propyl N,N-dipropyl thio-
carbamate
The method described below is primarily that presently used by
the State of Virginia but written into our standard format; however,
it is followed by a different set of conditions from a tentative EPA
method.
Reagents;
1. Vernolate standard of known % purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
2 Vernolate EPA-1
Equipment:
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.1 ran NaCl or KBr cells
*
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*
4. Usual laboratory glassware
* Virginia laboratories place their weighed samples
in 25 mm x 200 mm screw-top culture tubes, add solvent
by pipette, put in 1-2 grams anhydrous sodium sulfate,
and seal tightly with teflon-faced rubber-lined screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Procedure:
Preparation of Standard:
Weigh 0.12 gram vernolate standard into a 10 ml volumetric
flask, make to volume with chloroform, and mix well. Add a
small amount of anhydrous sodium sulfate to insure dryness.
(final cone 12 mg/ml)
Preparation of Sample:
For emulsifiable concentrates, weigh a portion of sample
equivalent to 0.6 gram vernolate into a 50 ml volumetric flask,
make to volume with chloroform, and mix well. Add a few grams
of anhydrous sodium sulfate to insure dryness and clarify the
solution, (final cone 12 mg vernolate/ml)
For granular formulations, weigh a portion of sample equiv-
alent to 0.6 gram vernolate into a glass-stoppered flask or
-------
3 Vernolate EPA-1
screw-cap bottle. Add 50 ml chloroform by pipette and 1-2 grams
anhydrous sodium sulfate. Close tightly and shake for one hour.
Allow to settle; centrifuge or filter if necessary, taking pre-
caution to prevent evaporation, (final cone 12 mg vernolate/ml)
Determination:
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1850 cm to
1500 cm'1 (5.4 p to 6.7 ;u) .
Determine the absorbance of standard and sample using the
peak at 1630 cm~ (6.13 p) and basepoint at 1800 cm (5.56 ;a) .
Calculations;
From the above absorbances and using the standard and sample
concentrations, calculate the percent vernolate as follows:
y _ (abs. sample) (cone, std in mg/ml)_(% purity std)
(abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg vernolate/ml chloroform gives an
absorbance of approx. 0.024 in a 0.1 mm cell.)
The above method was contributed by the Commonwealth of Virginia,
Division of Consolidated Laboratory Services.
See EPA method on page 4.
-------
4 Vernolate EPA-1
The conditions below are those used in a tentative EPA method —
method developed by George Radan, EPA Region II, New York.
Procedure: same as described above
Solvent: carbon disulfide
Concentration of standard: 6 mg/ml
Concentration of sample: equivalent to 6 mg vernolate/ml
IR cell: 0.5 mm
Scan range: 1250 cm"1 to 950 cm"1 (8.0 p. to 10.5 p)
Analytical peak: 1105 cm"1 (9.05 Ju)
Baseline: 1163 cm"1 to 1047 cm"1 (8.6 p to 9.55 p)
Calculation: same
-------
October 1975 Vernolate EPA-2
Determination of Vernolate
by Gas-Liquid Chromatography
(FID - Internal Standard)
Vernolate is the common name for S-propyl dipropylthiocarbamate,
a registered herbicide having the chemical structure:
CH2—CH2 CH3
CH3—CH2—CH2—S—C N
— CH2—CH3
Molecular formula: C H NOS
Molecular weight: 203.4
Boiling point: 140°C at 20 mm Hg, 150°C at 30 mm Hg
Physical state, color, and odor: clear liquid with an aromatic odor
Solubility: about 100 ppm in water at 20-21°C; miscible with
common organic solvents
Stability: stable; non-corrosive
Other names: Vernam (Stauffer), R-1607, S-propyl N,N-dipropyl thio-
carbamate
Reagents:
1. Vernolate standard of known % purity
2. Cycloate standard of known % purity
3. Carbon disulfide, pesticide or spectro grade
-------
2 Vernolate EPA-2
Reagents (Cont.):
4. Chloroform, pesticide or spectro grade
5. Methanol, pesticide or spectro grade
6. Internal Standard solution - weigh 0.20 gram cycloate into
a 50 ml volumetric flask? dissolve in and make to volume
with a solvent mixture consisting of 80% carbon disulfide
+ 15% chloroform + 5% methanol. (cone 4 mg cycloate/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 4 mm glass column packed with 3% OV-1 on
60/80 Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 jal
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID:
Column temperature: 140°C
Injection temperature: 225°C
Detector temperature: 250°C
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi
Hydrogen pressure: 20 psi
Air pressure: 30 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
3 Vernolate EPA-2
Procedure:
Preparation of Standard:
Weigh 0.08 gram vernolate standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 20 ml of
the internal standard solution and shake to dissolve. (final
cone 4 mg vernolate and 4 mg cycloate/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.08 gram vernolate
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the vernolate. For
coarse or granular materials, shake mechanically for 30 minutes
or shake by hand intermittently for one hour. (final cone 4 mg
vernolate and 4 mg cycloate/ml)
Determination:
Inject 2-3 pi of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from
1/2 to 3/4 full scale. The elution order is vernolate, then
cycloate.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of vernolate and cycloate
from both the standard-internal standard solution and the sample-
internal standard solution.
-------
4 Vernolate EPA-2
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
VJT? _ (wt. cycloate)(% purity cycloate) (pk. ht. or area vernolate)^
(wt. vernolate)(% purity vernolate)(pk. ht. or area cycloate)
Determine the percent vernolate for each injection of the
sample-internal standard solution as follows and calculate the
average:
7 _ (wt. cycloate)(% purity cycloate)(pk. ht. or area vernolate)(100)
(wt. sample) (pk. ht. or area cycloate) (RF) (^~~l)
Method submitted by Division of Regulatory Services, Kentucky
Agricultural Experiment Station, University of Kentucky, Lexington,
Kentucky 40506.
-------
October 1975 Vernolate EPA-3
(Tentative)
Determination of Vernolate
by Gas-Liquid Chromatography
(TCD - Internal Standard)
Vernolate is the common name for S-propyl dipropylthiocarbamate,
a registered herbicide having the chemical structure:
0
CH2—CH2—CH3
CH3 — CH2—CH2 — S — C >.
CH2—CH3
Molecular formula: C H NOS
Molecular weight: 203.4
Boiling point: 140°C at 20 mm Hg, 150°C at 30 mm Hg
Physical state, color, and odor: clear liquid with an aromatic odor
Solubility: about 100 ppm in water at 20-21°C; miscible with common
organic solvents
Stability: stable; non-corrosive
Other names: Vernam (Stauffer), R-1607, S-propyl N,N-dipropyl thio-
carbamate
Reagents:
1. Vernolate standard of known % purity
2. Butylate standard of known % purity
3. Carbon disulfide, pesticide or spectro grade
-------
2 Vernolate EPA-3
(Tentative)
Reagents (Cont.):
4. Chloroform, pesticide or spectro grade
5. Acetone, pesticide or spectro grade
6. Internal Standard solution - weigh 0.25 gram butylate
standard into a 25 ml volumetric flask, dissolve in, and
make to volume with a solvent mixture consisting of 80%
carbon disulfide H- 15% chloroform + 5% acetone. (final
cone 10 mg butylate/ml)
Equipment:
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 5' x 1/4" glass column packed with 5% PEG-1540
on 60/80 mesh Chromosorb W AW DMCS (or equivalent
column)
3. Precision liquid syringe: 25 or 50 pi
4. Mechanical shaker
5. Usual laboratory glassware
Operating Conditions for TCI):
Column temperature: 150°
Injection temperature: 200°
Detector temperature: 175°
Filament current: 200 ma
Carrier gas: Helium
Carrier gas flow rate: 30 ml/min
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Vernolate EPA-3
(Tentative)
Procedure:
Preparation of Standard;
Weigh 0.1 gram vernolate standard into a small glass-stoppered
flask or screw-cap tube. Add by pipette 10 ml of the internal
standard solution and shake to dissolve, (final cone 10 mg verno-
late and 10 mg butylate/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.1 gram vernolate
into a small glass-stoppered flask or screw-cap tube. Add by
pipette 10 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the vernolate. For
coarse or granular materials, shake or tumble mechanically for
30 minutes or shake by hand intermittently for one hour, (final
cone 10 mg vernolate and 10 mg butylate/ml)
Determination:
Inject 10-15 jul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from
1/2 to 3/4 full scale. The elution order is butylate, then
vernolate.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation:
Measure the peak heights or areas of vernolate and butylate
from both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
-------
Vernolate EPA-3
(Tentative)
. butylate) (% purity butylate) (pk. ht . or area vernolate)
(wt. vernolate) (% purity vernolate) (pk. ht. or area butylate)
Determine the percent vernolate for each injection of the
sample-internal standard solution as follows and calculate the
average:
7 - lwtv butyJLate) (% purity butylate) (pk. ht. or area vernolate) Q&&)~~
(wt. sample) (pk. ht. or area butylate) (RF) ftf-lj
This method was submitted by the Commonwealth of Virginia, Division
of Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.
Note! This method has been designated as tentative since it is a
Va. Exp. method and because some of the data has been suggested
by EPA's Beltsville Chemistry Lab. Any comments, criticism,
suggestion, data, etc. concerning this method will be appreciated,
-------
November 1975
Warfarin EPA-1
(Tentative)
Determination of Warfarin by
High Pressure Liquid Chromatography
Warfarin is the official common name for 3-(alpha-acetonylbenzyl)-
4-hydroxycoumarin, a registered rodenticide having the chemical struc-
ture:
OH
C=0
Molecular formula: C,.H,,0. CH3
19 16 4
Molecular weight: 308.3
Melting point: (dl form) 159 to 161°C
Physical state, color, odor, taste: (dl form) colorless, tasteless,
odorless crystals
Solubility: practically insoluble in water and benzene, moderately
soluble in alcohols, readily soluble in acetone and
dioxane; forms water-soluble salts with sodium
Stability: stable under normal conditions
Other names: WARF (Wisconsin Alumni Research Foundation), coumafene
(France), zoocoumarin (Netherlands, USSR), Kypfarin
-------
2 Warfarin EPA-1
(Tentative)
Reagents;
1. Warfarin standard of known % purity
2. Methanol, pesticide or spectro grade
3. Phosphorous acid solution, 0.0025M in water
4. Dioxane, pesticide or spectro grade
Equipment;
1. High pressure liquid chromatograph with UV detector at 254 run.
If a variable wavelength UV detector is available, other wave-
lengths may be useful to increase sensitivity or eliminate
interference. Warfarin is more easily determined at 308 nm.
2. Suitable column such as:
a. DuPont ODS Permaphase, 1 meter x 2.1 mm ID
b. Perkin-Elmer ODS Sil-X 11 RP, 1/2 meter x 2.6 mm ID
3. High pressure liquid syringe or sample injection loop
4. Usual laboratory glassware
Operating Conditions;
Mobile phase: 10% methanol + 90% 0.0025M H^O^ in water
Column temperature: 50"C
Chart speed: 5 min/inch or equivalent
Flow rate: 0.5 to 1.5 ml/min (Perkin-Elraer 1/2 meter column)
Pressure: 500 psi (DuPont 1 meter column)
Attenuation: Adjusted
Conditions may have to be varied by the analyst for the specific
instrument being used, column variations, sample composition, etc. to
obtain optimum response and reproducibility.
-------
Warfarin EPA-1
(Tentative)
Procedure:
Preparation of Standard:
Weigh 0.05 gram warfarin standard into a 50 ml volumetric
flask; dissolve in and make to volume with dioxane. Mix
thoroughly, pipette 5 ml into a second 50 ml volumetric flask,
make to volume with dioxane, and mix well, (final cone 0.1 mg/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.005 gram warfarin
into a glass-stoppered or screw-cap 125 ml Erlenmeyer flask, add
50 ml dioxane by pipette, close tightly, and shake for one hour.
Allow to settle; centrifuge or filter if necessary, taking pre-
caution to avoid evaporation, (final cone 0.1 mg warfarin/ml)
Determination;
For a variable wavelength detector, use 308 nm rather than
254 nm. Warfarin is more easily detected at this wavelength and
many interferences are eliminated or reduced to a negligible
amount.
Alternately inject three 5 fii portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the percent
warfarin as follows:
(pk.
(Pk7
ht. or area sample)(wt. std injected)(% purity of std)
pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA, Beltsville, Md.
-------
November 1975
Warfarin EPA-2
Determination of Warfarin
by Ultraviolet Spectroscopy
Warfarin is the official common name for 3-(alpha-acetonylbenzyl)-
4--hydroxycoumarin, a registered rodenticide having the chemical struc-
ture:
OH
H
c=o
Molecular formula: C,_H,,0,
19 16 4
Molecular weight: 308.3
Melting point: (dl form) 159 to 161°C
Physical state, color, odor, taste: (dl form) colorless, tasteless,
odorless crystals
Solubility: practically insoluble in water and benzene, moderately
soluble in alcohols, readily soluble in acetone and
dioxane; forms water-soluble salts with sodium
Stability: stable under normal conditions
Other names: WARF (Wisconsin Alumni Research Foundation), coumafene
(France), zoocoumarin (Netherlands, USSR), Kypfarin
This method is applicable to most bait materials containing about
0.025% warfarin or its sodium salt. It is especially useful for bait
materials that have a glazed coating or that have been made into pellets.
-------
2 Warfarin EPA-2
In such cases the extraction of warfarin in organic solvents (AOAC 12th Ed,
6.140-6.141 ether extraction) is retarded.
Reagents :
1. Warfarin standard of known % purity
2. Sodium pyrophosphate, 1% solution - dissolve 5 grams
Na.P00_.10H00 in water and make to 500 ml.
427 2
3. Ether-hexane mixture - extract 200 ml n-hexane (bp 60 — 68°C)
with three 20 ml portions of 1% pyrophosphate solution and
add 50 ml ethyl ether, making a 20% ether-80% hexane mixture.
4. Hydrochloric acid, 2.5N solution - 20.6 ml hydrochloric acid
diluted to 100 ml.
Equipment:
1. Ultraviolet spectrophotometer , double beam ratio recording
with matched 1 cm quartz cells
2. Centrifuge with 50 ml and 100 ml glass-stoppered tubes
3. Mechanical shaker
4. Usual laboratory glassware
Procedure :
of Standard :
Weigh 0.1 gram warfarin standard into a 100 ml volumetric
flask; dissolve in and make to volume with 1% sodium pyrophosphate.
Mix thoroughly, pipette 10 ml into a second 100 ml volumetric flask,
and make to volume with 1% pyrophosphate solution. Again, mix
thoroughly, pipette 10 ml of this solution into a third 100 ml
volumetric flask, and make to volume with the 1% pyrophosphate
solution. (final cone 10 jug/ml)
-------
3 Warfarin EPA-2
Preparation of Sample:
Weigh a portion of sample equivalent to 0.0005 gram warfarin
(2 grams for a 0.025% product) into a 125 ml glass-stoppered
Erlenmeyer flask, add by pipette 50 ml 1% pyrophosphate solution,
close tightly, and shake on a mechanical shaker for one hour.
Transfer 30-35 ml to a glass-stoppered centrifuge tube and centri-
fuge for 5 minutes. Pipette 25 ml of clear solution into a second
centrifuge tube, add 5 ml 2.5N hydrochloric acid and 50 ml ether-
hexane solution, stopper tightly, and shake for 5 minutes. If an
emulsion forms, centrifuge a few minutes to break the emulsion.
Pipette 20 ml of the ether layer into another centrifuge tube
and add by pipette 10 ml 1% pyrophosphate solution. Shake for 2
minutes and remove the ether layer — this is conveniently done by
using a tube drawn into a fine tip and connected to a water
aspirator. If the aqueous phase is not clear, centrifuge for a
few minutes with the top off to remove any traces of the ether-
hexane phase. (final cone 10 jig warfarin/ml)
HY Petermination:
With the UV spectrophotometer at the optimum quantitative
settings, balance the 0 and 100% at 308 nm with the 1% pyrophos-
phate solution in each cell. Scan both standard and sample from
360 nm to 240 nm, using the 1% pyrophosphate solution in the
reference cell. I'easure the absorbance of both standard and
sample at 308 nm.
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent warfarin as follows:
_ J[abs_. sampj.e)_(cojic._ std in jug/ml) (% purity std)
(abs. std)(cone, sample in pg/ml)
-------
December 1975
Warfarin EPA-3
(Tentative)
Determination of Warfarin, Sodium Salt
by High Pressure Liquid Chromatography
Warfarin is the official common name for 3-(alpha-acetonylbenzyl)-
4-hydroxycoumarin, a registered rodenticide having the chemical struc-
ture:
c=o
Molecular formula: C,nH,,0.
19 16 4
Molecular weight: 308.3
Melting point: (dl form) 159 to 161°C
Physical state, color, odor, taste: (dl form) colorless, tasteless,
odorless crystals
Solubility: practically insoluble in water and benzene, moderately
soluble in alcohols, readily soluble in acetone and
dioxane; forms water-soluble salts with sodium
Stability: stable under normal conditions
Other names: WARF (Wisconsin Alumni Research Foundation), coumafene
(France), zoocoumarin (Netherlands, USSR), Kypfarin
-------
Warfarin EPA-3
(Tentative)
Reagents;
1. Warfarin standard oE known % purity
2. Methanol, pesticide or spectro grade
3. Phosphorous acid solution, 0.0025M in water
4. Sodium pyrophosphate, 1% solution - dissolve 10 grains
Na.P.O-.lOH.O in water and make to 1000 ml.
427 2
Equipment:
1. High pressure liquid chromatograph with UV detector at 254 nm.
If a variable wavelength UV detector is available, other wave-
lengths may be useful to increase sensitivity or eliminate
interference. Warfarin is more easily determined at 308 nm.
2. Suitable column such as:
a. DuPont ODS Permaphase, 1 meter x 2.1 mm ID
b. Perkin-Elmer ODS Sil-X 11 RP, 1/2 meter x 2.6 mm ID
3. High pressure liquid syringe or sample injection loop
4. Usual laboratory glassware
Operating Conditions;
Mobile phase:
Column temperature:
Chart speed:
Flow rate:
Pressure:
Attenuation:
10% methanol + 90% ,0.0025M H PO^, in water
50°C
5 min/inch or equivalent
0.5 to 1.5 ml/min (Perkin-Elmer 1/2 meter column)
500 psi (DuPont 1 meter column)
Adjusted
Conditions may have to be varied by the analyst for the specific
instrument being used, column variations, sample composition, etc. to
obtain optimum response and reproducibility.
-------
3 Warfarin EPA-3
(Tentative)
Procedure;
Preparation of Standard;
Weigh 0.05 gram warfarin standard into a 50 ml volumetric
flask; dissolve in and make to volume with sodium pyrophosphate
solution. Mix thoroughly, pipette 5 ml into a second 50 ml
volumetric flask, make to volume with sodium pyrophosphate solution,
and mix well, (final cone 0.1 mg/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.005 gram warfarin
into a glass-stoppered or screw-cap 125 ml Erlenmeyer flask, add
50 ml sodium pyrophosphate solution by pipette, close tightly, and
shake for one hour. Allow to settle; centrifuge or filter if
necessary, taking precaution to avoid evaporation, (final cone
0.1 mg warfarin/ml)
Determination:
For a variable wavelength detector, use 308 nm rather than
254 nm. Warfarin is more easily detected at this wavelength and
many interferences are eliminated or reduced to a negligible
amount.
Alternately inject three 5 pi portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the percent
warfarin as follows:
„ „ (pk. ht. or area sample) (wt. std injected) (% purity of std)
(pk. ht. or area standard)(wt. sample injected)
% Sodium salt of warfarin * 1.071 x % warfarin
Method submitted by Elmer H. Hayes, EPA, Beltsville, Md.
-------
-------
December 1975
Zinc Phosphide EPA-1
Determination of Zinc Phosphide
by the Phosphine Evolution Method
Zinc phosphide is a registered rodenticide having the chemical
structure:
Molecular formula: Zn_P?
Molecular weight: 258.1
Melting point: 420°C (sublimes when heated in the absence of oxygen)
Physical state, color, and odor: gray powder, disagreeable odor (not
offensive to rodents)
Solubility: practically insoluble in water and ethanol; soluble in
benzene and carbon disulfide
Stability: stable when dry but decomposes slowly in moist air; reacts
violently with acids with decomposition to the spontaneously
inflammable phosphine
Other names: Kilrat, Mous-con, Rumetan
Principle of the Method:
A weighed portion of sample is initially washed with distilled
water to remove any antimony potassium tartrate which would interfere
with the quantitative evolution of phosphine from zinc phosphide. The
X
-------
2 Zinc Phosphide EPA-1
washed sample is treated with sulfuric acid under an atmosphere of nitro-
gen to release phosphine gas which is swept by the nitrogen into several
absorption flasks containing standard potassium permanganate solution
with which it reacts. The excess permanganate is titrated with oxalic
acid solution and the zinc phosphide calculated from the amount of
permanganate used by the phosphine from the sample.
The antimony potassium tartrate in the wash solution may be deter-
mined by titration with iodine solution.
Reagents:
1. Potassium permanganate, 0.5N standard solution - 15.81 grams
KMn04 per liter
2. Sulfuric acid, 10% solution - 1 volume concentrated sulfuric
acid added to 9 volumes water
3. Oxalic acid, 0.5N standard solution - 31.52 grams (COOH) ,2H 0
per liter. This solution should contain 125 to 150 ml concen-
trated sulfuric acid.
4. Distilled water - freshly boiled and cooled to 15°C
5. Nitrogen gas
6. Sodium bicarbonate, saturated solution
7. Starch indicator solution
8. Iodine, 0.1N standard solution
Equipment:
Reaction train consisting of a 500 ml Erlenmeyer flask fitted
with a three-hole stopper for: (1) an inlet tube for nitrogen,
(2) a separatory funnel for adding acid, and (3) an outlet tube
leading to three absorption flasks, each with an inlet tube
extending to the bottom of the flask and an outlet tube leading
to the next flask. It is very convenient to have the flasks
connected with polyethylene tubing and polyethylene friction
connectors.
-------
Zinc Phosphide EPA-1
C${J3
2. Water bath maintained at 50"C
3. Titrating equipment
A. Usual laboratory glassware
Procedure:
Preparation of Sample;
Weigh a portion of ground sample equivalent to 0.005-0.010
gram zinc phosphide into a 250 ml beaker. Add 50-75 ml freshly
boiled and cooled distilled water, stir, and filter with gentle
suction through ashless, double acid washed filter paper. Trans-
fer all the sample into the paper and wash five times with 15 ml
portions of distilled water.
Use the residue for the determination of zinc phosphide and
the filtrate for the determination of antimony potassium tartrate.
Determination of Antimony Potassium Tartrate:
If antimony potassium tartrate is to be determined, add 10 ml
cold saturated solution of sodium bicarbonate and a few drops of
0.5% starch indicator solution to the combined filtrate and
-------
4 Zinc Phosphide EPA-1
titrate immediately with 0.1N iodine solution to a permanent blue
color. Calculate the % antimony potassium tartrate as follows:
% „ (ml iodine)(N iodine)(0.1625)(100)
(wt. sample)
Evolution and Absorption of Phosphine;
Transfer the filter paper and residue (from above) to the
500 ml reaction flask. Pipette 100 ml of 0.5N standard potassium
permanganate into the first absorption flask, and pipette 50 ml
into each of the other two. Add 100 ml of 10% sulfuric acid to
the separatory funnel, connect the apparatus to a source of
nitrogen, sweep the system with nitrogen for at least 10 minutes,
and adjust the flow of nitrogen to one or two bubbles per second.
Slowly add the acid to the reaction flask, regulating the rate so
that a steady stream of bubbles appears in the absorbers. After
all the acid has been added, place the reaction flask in the 50°C
water bath and allow the reaction to continue for at least one
hour, adjusting the flow of nitrogen to maintain a steady flow of
bubbles at all times.
Determination of Phosphine and Calculation of Zinc Phosphide;
At the end of the reaction period, quantitatively transfer the
potassium permanganate solution from the absorbers into a one-liter
beaker. Accurately measure 225 ml of the 0.5N oxalic acid standard
solution into a plastic squeeze wash bottle and rinse the absorbers
and connecting tubes into the liter beaker. Carefully dissolve all
the manganese dioxide and rinse with distilled water so as not to
lose any of the oxalic acid solution. Finally, rinse the oxalic
acid solution from the wash bottle into the same liter beaker.
-------
5 Zinc Phosphide EPA-1
Warm the oxalic-manganous solution to about 50°C and titrate
the excess oxalic acid with the 0.5N potassium permanganate
solution to the first permanent pink (persists for 60 seconds).
Calculate the zinc phosphide as follows:
meq's from KMnO, «• N x (ml KMnO, added 4- ml used in titration)
meq_'s_ from oxalic acid a N x ml oxalic acid used
meq's difference * net meq's used by sample
% zinc phosphide
r
(net meq's) (0.01613) (100)
(grams of sample)
milliequivalent weight of zinc phosphide is 0.01613 or
Reactions:
3PH 4- 8KMnO.
3
(COOH)
> 8MnO> + K0HPO. + 2K.PO. + 4H.O
224342
(COOH)0 -f 2KMnO. + 3H.SO.
2 424
MnSO, 4- 2C02t + 2H 0
K.SO. + 2MnSO. + 10 C00"t 4- 8H00
24 4 2 2
-------
December 1975
Zinc Phosphide EPA-2
Determination of Zinc Phosphide in Grain Baits
by Gas-Liquid Chromatography (FPD)
Zinc phosphide is a registered rodenticide having the chemical
structure:
Molecular formula: Zn-ipo
Molecular weight: 258.1
Melting point: A20°C (sublimes when heated in the absence of oxygen)
Physical state, color, and odor: gray powder, disagreeable odor (not
offensive to rodents)
Solubility: practically insoluble in water and ethanol; soluble in
benzene and carbon disulfide
Stability: stable when dry but decomposes slowly in moist air; reacts
violently with acids with decomposition to the spontaneously
inflammable phosphine
Other names: Kilrat, Mous-con, Rumetan
Reagents;
1. Zinc phosphide standard of known % purity
2. Glucose, 100 mesh, dry powder
3. Toluene, pesticide or spectre grade
4. Sulfuric acid, 10% solution
-------
2 Zinc Phosphide EPA-2
Equipment:
1. Gas chromatograph with flame photometric detector (FPD) and
phosphorus filter (526 nm emission band)
2. Column: 4' x 1/4" O.D. glass column packed with 5% QF-1 on
80/100 mesh Gas Chrom Q, conditioned isothermally
at 40-50°C (or equivalent column)
3. Precision liquid syringe: 10 fil
4. Electric sample mill or blender
5. Ultrasonic cleaner (aid to dispersion and dissolution of samples)
6. Usual laboratory glassware
Operating Conditions for FPD:
Column temperature: 40-50°C
Injection temperature: 200°C
Detector temperature: 140-150°C
Nitrogen carrier gas: 45-60 ml/rain
Hydrogen to Detector: 50-150 ml/min
Air to Detector: 0-35 ml/min
Oxygen to Detector: 10-25 ml/min
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
The linear detector response for phosphine should be determined at
the concentrations of interest. Reference standards must be prepared each
day as phosphine is not stable for prolonged periods. Analyses are
referred to the reference standards rather than to a prepared standard
curve.
-------
3 Zinc Phosphide EPA-2
Procedure;
Preparation of Standard:
Prepare a 1% mixture of zinc phosphide in glucose as follows:
weigh 1.00 gram zinc phosphide (correcting for less Chan 100%
purity) and make to 100 grams with dry, powdered glucose; mix
thoroughly to insure a homogenous mixture.
*
Weigh 0.38 gram of this diluted standard mixture into a 100 ml
volumetric flask and fill to the mark with toluene. Add sufficient
10% sulfuric acid solution to bring the liquid level within 1 cm
of the bottom of the glass stopper. Set aside for one hour, mixing
occasionally by inverting several times and shaking for one minute.
Seal the top with tape to prevent loss of toluene and place
in an ultrasonic bath for five minutes. Remove and let stand
another hour. Optimum hydrolysis and absorption into the toluene may
be a-chievedby allowing the hydrolyzed samples to stand overnight.
Standards and samples can be kept 24 hours if a toluene-to-
glass seal is made by tilting the flask to cover the stopper.
(final cone approx 10 ppm PH» or 10 pg PH_/ml)
(0.3794 g of the 1% mixture will hydrolyze to 1 mg phosphine,
which, dissolved in 100 ml toluene, gives a 10 jug/ml cone)
Preparation of Sample:
Chill a blender or an electric sample mill in a freezer until
well chilled, add 30-40 grams of grain bait sample, and grind to a
flour-like powder. Weigh 0.38 gram (for 1% formulation, 0.19 g for
2% formulation) into a 100 ml volumetric flask and follow the same
procedure as above under preparation of standard, (final cone same
as standard)
-------
Zinc Phosphide EPA-2
Determination;
Using a precision liquid syringe, inject 1 ul of standard
solution and adjust attenuation to a 30-50% full scale response.
Inject 1 ul of sample solution using the same conditions. When
the peak heights for both the sample and standard are reproducible
within + 5%, make alternate injections of sample and standard.
Measure the peak heights in mm of the standard and sample.
Calculation:
From the average peak heights of standard and sample,
calculate the percent zinc phosphide as follows:
°j „ (pk. ht. sample)(wt. std injected) (100)
(pk. ht. std)(wt. sample injected)
Any deficiencies found in formulations by this method should
be checked by method EPA-1 (phosphine evolution method).
Method submitted by the Hawaiian Sugar Planters' Association, 1527
Keeaumoku Street, Honolulu, Hawaii 96822.
-------
-------
UPDATE 1
EPA MANUAL OF CHEMICAL METHODS FOR PESTICIDES
AND DEVICES
JUH 2 5 1537
-------
-------
Dear Subscriber:
Enclosed is the first update of the EPA Manual of Chemical
Methods for Pesticides and Devices. This update includes:
1. Nineteen additional methods for the analysis of commercial
pesticide formulations
2. One analytical method for the degradation product "ethylene-
thiourea" in ethylenebisdithiocarbamate fungicide formula-
tions
3. Pesticide Name Cross Reference Index to the above 20 methods
4. "Pen and ink" corrections to 59 methods
5. Special major correction to the Diphacinone EPA-1 method
A second update for this manual is tentatively scheduled for
January 1978 and will include additional analytical methods,
revisions, and corrections.
The Editorial Committee would appreciate written comments in
relation to the following:
1. Corrections or modifications in data, analytical procedures,
or calculations in the methods now in the manual
2. New methods or data for inclusion in future updates or
revisions of this manual
3. Suggestions for additional methods, graphs, charts, data,
or information (general or specific) that would increase
the usefulness of this manual
Such comments may be made to members of the Editorial Committee
or the AAPCO-EPA Review Committee as listed in the Preface (page 4)
or sent to Jack B. Looker, Assistant Chairman, or Warren R. Bontoyan,
Chairman, Editorial Committee.
Address: EPA, TSD
Room 101, Bldg. 306, ARC-East
Beltsville, Md. 20705
Warren R. Bontoyan
Chairman, Editorial Committee
Jack B. Looker
Asst. Chairman, Editorial Committee
-------
-------
Diphacinone EPA-1 - Special Correction
The method "Diphacinone EPA-1, Determination of Diphacinone
in Baits by Ultraviolet Spectroscopy, November 1975" is no
longer satisfactory for the analysis of diphacinone. Commercial
bait formulations are more complex, including meat, fish, and
apple flavors. This necessitates a more thorough extraction
procedure such as in the following method.
Changes to be made are as follows;
Change November 1975 to: July 1977
(Revision of November 1975)
Add (tentative) to Diphacinone EPA-1
') (3) Replace pages 2 and 3 with the following method
-------
-------
Analytical Methods - First Supplement
July 1, 1977
Errors to be corrected;
U-l) 4-Aminopyridine EPA-1 (tentative^
Pg 2 under UV Determination, 3rd and last lines
"302" should be "262"
:j(--/; Chlorophenoxy Herbicides EPA-1
The "second page 7 (containing Erbon data)" should be
"page 8" and "page 8" should be "page 9" **
.'• Chlorophenoxy Herbicides EPA-2
Pg 3 under UV Determination, middle of line 4
"286" should be "296" **
i- * I) Norbormide EPA-1
Pg 2 and 3, top right corner
Change "EPA-2" to "EPA-1" **
u^ \] Qrganotin Compounds EPA-1
Pg 5, calculation of % tin should be
-, „. (ml KIOJ(N KIO,) (0.05935) (100)
/o cm ** j j
(grams sample)
M-l\ Pebulate EPA-1 (tentative)
Pg 3 under Note!
"iso-publate" should be "iso-pebulate" **
^- j ] Phenols and Chlorophenols EPA-1
Pg 3, bottom line
"Chlorophene" should be "Clorophene" **
{A~] Pyrethrins EPA-2
Pg 1 under Equipment; 2.
column packing is 60-80 mesh **
' Ronnel EPA-2
Pg 3 under Preparation of Standard;
"EPIC" should be "ronnel" **
'- ?' f)Strychnine EPA-1
Pg 2 under Reagents; 1.
"ethanol" should be "ether"
** Manuals distributed by AOAC were corrected before publication
-------
49 GLC internal standard methods listed below
Under Calculation, the factor "(100)" should be deleted
from the "% - " calculation.
This applies to the following methods:
chlor EPA-1 (tentative) //-/^esmethrin EPA-5 (tentative)
Alachlor EPA-2 (tentative) ^-/feonnel EPA-2
^Anilazine EPA-2 (tentative) ,^-^Terbutol EPA-2 (tentative)
^Atrazine EPA-2 (tentative) '(^Trifluralin EPA-1
JJenefin EPA-2 (tentative) ^-l)Vernolate EPA-2
^romacil EPA-1 (tentative) ^-;> Vernolate EPA-3 (tentative)
t^utylate EPA-4
^Butylate EPA-5 (tentative)
yChlorophenoxy Herbicides EPA-4 (tentative)
t Chlorophenoxy Herbicides EPA-5 (tentative)
^Chloroxuron EPA-2 (tentative)
,/Coumaphos EPA-3 (tentative)
i/Crufomate EPA-2 (tentative)
•/Cycloate EPA-3
,^Deet EPA-2 (tentative)
,Deet EPA-3 (tentative)
i/Diazinon EPA-4
^p-Dichlorobenzene EPA-2 (tentative)
^isulfoton EPA-2 (tentative)
^/Endosulfan EPA-3 (tentative)
v/Endosulfan EPA-4 (tentative)
^EPTC EPA-1 (tentative)
,/EPTC EPA-3
,/EPTC EPA-4 (tentative)
j^EPTC EPA-5 (tentative)
yEthoprop EPA-2 (tentative)
t^Ethoprop EPA-3 (tentative)
t/Ethyl Hexanediol EPA-2 (tentative)
^Metaldehyde EPA-2 (tentative)
j/Methoxychlor EPA-2 (tentative)
v/Methyl Parathion EPA-4
^Methyl Parathion EPA-5
t^ietobromuron EPA-3 (tentative)
4/Monocrotophos EPA-2
/Parathion EPA-2 (tentative)
i/Pebulate EPA-2 (tentative)
/Pebulate EPA-3 (tentative)
i/Phenols & Chlorophenols EPA-8 (tentative)
,/Piperonyl Butoxide EPA-2
^rometone EPA-1 (tentative)
^Prometone EPA-2 (tentative)
,/Propargite EPA-2 (tentative)
-i) Resmethrin EPA-3 (tentative)
The Editorial Staff of this manual would appreciate hearing of
other errors so that they may be corrected in future updates.
-------
ANALYTICAL METHODS - FIRST SUPPLEMENT
July 1, 1977
Antimycin A EPA-1 (tentative)
Asulam EPA-1 (tentative)
Bentazon EPA-1 (tentative)
Chlorobenzilate EPA-1 (tentative)
Chlorophacinone EPA-1 (tentative)
Crotoxyphos EPA-1 (tentative)
Diraethoate EPA-1 (tentative)
Dimethoate EPA-2 (tentative)
Ethylenethiourea EPA-1 (tentative)
Linuron EPA-3 (tentative)
Mercaptobenzothiazole EPA-1 (tentative)
Mercaptobenzothiazole EPA-2 (tentative)
Methidathion EPA-1 (tentative)
Monocrotophos EPA-3 (tentative)
Propylene Glycol EPA-1 (tentative)
Trichlorfon EPA-1 (tentative)
Trichlorfon EPA-2 (tentative)
Triethylene Glycol EPA-1 (tentative)
Vacor (trade name) EPA-1 (tentative)
Vacor (trade name) EPA-2 (tentative)
-------
-------
ANALYTICAL METHODS - FIRST SUPPLEMENT
July 1, 1977
Pesticide Name Cross Reference Index to the Methods
Acaraben
Afalon
Akar
Antimycin A EPA-1 (tentative)
Asulam EPA-1 (tentative)
Asulame
Asulox
Azodrin
Bantex (zinc salt)
Basagran
Bayer 15922
Bayer L 13/59
Bentazon EPA-1 (tentative)
Benzilan
C-23992
Caid
Captax
Chlorobenzilate EPA-1 (tentative)
Chlorofos
Chlorophacinone EPA-1 (tentative)
2-kp-chlorophenyl)phenylacety]J -
i^ 3-indandione
Chlorobenzilate EPA-1
Linuron EPA-3
Chlorobenzilate EPA-1
UV
uv
Asulam EPA-1
Asulam EPA-1
Monocrotophos EPA-3
Mercaptobenzothiazole EPA-1 & 2
Bentazon EPA-1
Trichlorfon EPA-1 & 2
Trichlorfon EPA-1 & 2
UV
Chlorobenzilate EPA-1
Chlorobenzilate EPA-1
Chlorophacinone EPA-1
Mercaptobenzothiazole EPA-1 & 2
GLC-FID-IS
Trichlorfon EPA-1 & 2
UV
Chlorophacinone EPA-1
-------
2-(2-p-chloropheny1-2-phenylac ety 1)
indane-1,3-dione
Ciodrin
Crotoxyphos EPA-1 (tentative)
Cygon
Chlorophacinone EPA-1
Crotoxyphos EPA-1
GLC-FID-IS
Dimethoate EPA-1 & 2
Daphene
De-Fend
Dermacid
3-(3,4-dichlorophenyl)-1-raethoxy-l-
methylurea
S-(2,3-dihydro-5-methoxy-2-oxo-l,3,4-
thiadiazol-3-yImethyl)dimethyl
phosphorothiolothionate
1,2-dihydroxypropane
Dimetate
Dimethoate EPA-1 (tentative)
Dimethoate EPA-2 (tentative)
Dimethogen
cis-3-(dimethoxyphosphinyloxy)-N-
methylcrotonamide
dimethyl-2-(alpha-methylbenzocarbonyl) •
1-methyl vinyl phosphate (E)
0, 0-dimethyl S- £(methylcarbamoyl)
methylj phosphorodithioate
0, O-dimethyl-O-(2-methylcarbamoyl-
1-methy1-viny1)-phosphate
dimethyl-l-methyl-2-methyl-
carbamoyl-vinyl phosphate
dimethyl cis-l-methyl-2-
(1-phenylethoxycarbonyl)vinyl
phosphate
Dimethoate EPA-1 & 2
Dimethoate EPA-1 & 2
Mercaptobenzothiazole EPA-1 & 2
Linuron EPA-3
Methidathion EPA-1
Propylene Glycol EPA-1
Dimethoate EPA-1 & 2
GLC-TCD-IS
GLC-FID-IS
Dimethoate EPA-1 & 2
Monocrotophos EPA-3
Crotoxyphos EPA-1
Dimethoate EPA-1 & 2
Monocrotophos EPA-3
Monocrotophos EPA-3
Crotoxyphos EPA-1
-------
dimethyl phosphate of alpha-
methylbenzyl 3-hydroxy-cis-crotonate
dimethyl phosphate of 3-hydroxy-N-
methyl-cis-crotonamide
0,0-dimethyl phosphorodithioate S-ester
with 4-(mercaptomethyl)-2-methoxy-
delta 2-l,3,4-thiadiazolin-5-one
dimethyl(2,2,2-trichloro-l-hydroxyethyl)
phosphonate
Dipterex
dipterex
Drat
Dylox
E. I. 12,880
ethyl 4,4*-dichlorodiphenylglycollate
ethyl 4,4'-dichlorobenzilate
2,2'-ethylenedioxybis (ethanol)
Ethylenethiourea EPA-1 (tentative)
ETU
Fintrol
Folbex
Postion MM
Crotoxyphos EPA-1
Monocrotophos EPA-3
Methidathion EPA-1
Trichlorfon EPA-1 & 2
Trichlorfon EPA-1 & 2
Trichlorfon EPA-1 & 2
Chlorophacinone EPA-1
Trichlorfon EPA-1 & 2
Dimethoate EPA-1 & 2
Chlorobenzilate EPA-1
Chlorobenzilate EPA-1
Triethylene Glycol EPA-1
GLC-FID & TCD
Ethylenethiourea EPA-1 (tentative)
Antimycin A EPA-1
Chlorobenzilate EPA-1
Dimethoate EPA-1 & 2
GS-13005
HOE 2810
Methidathion EPA-1
Linuron EPA-3
2-imidazolidinethione
3-isopropyl-lH-2,l,3-benzothiadiazin-
4(3H)-one 2,2-dioxide
Ethylenethiourea EPA-1
Bentazon EPA-1
-------
Kop-Mite
Chlorobenzilate EPA-1
L 395
Linuron EPA-3 (tentative)
Liphadione
Lorox
MB 9057
MET
Mercaptobenzothiazole EPA-1 (tentative)
Mercaptobenzothiazole EPA-2 (tentative)
2-mercaptobenzothiazole
Mertax
Methidathion EPA-1 (tentative)
S- [(5-methoxy-2-oxo-l,3,4-thiadiazol-
3(2H)-yl)methyl] 0,0-dimethyl
phosphorodithioate
methyl(4-aminobenzenesulphonyl)
carbamate
1-methylbenzyl 3-(dimethoxyphosphinyloxy)
-cis-crotonate
methylene glycol
methyl glycol
methyl sulfanilylcarbamate
metrifonate
Monocron
Monocrotophos EPA-3 (tentative)
Neguvon
Niacides
Dimethoate EPA-1 & 2
UV
Chlorophacinone EPA-1
Linuron EPA-3
Asulam EPA-1
Mercaptobenzothiazole EPA-1 & 2
UV
potentiometric titration
Mercaptobenzothiazole EPA-1 & 2
Mercaptobenzothiazole EPA-1 & 2
GLC-FID-IS
Methidathion EPA-1
Asulam EPA-1
Crotoxyphos EPA-1
Propylene Glycol EPA-1
Propylene Glycol EPA-1
Asulam EPA-1
Trichlorfon EPA-1 & 2
Monocrotophos EPA-3
GLC-FID-IS
Trichlorfon EPA-1 & 2
Mercaptobenzothiazole EPA-1 & 2
-------
Nuodex 84 (sodium salt)
Nuvacron
Partox
Perfekthion
Poast
1,2-propanediol
Propylene Glycol EPA-1 (tentative)
N-3-pyridylmethyl-N'-p-nitrophenylurea
Quick
Raviac
Rebelate
RH-787
Roger
Roxion
Rozol
Sarclex
SD 4294
Supracide
Thiotax
Trichlorfon EPA-1 (tentative)
Trichlorfon EPA-2 (tentative)
trichlorphon
Triethylene Glycol EPA-1 (tentative)
Mercaptobenzothiazole EPA-1 & 2
Monocrotophos EPA-3
Chlorophacinone EPA-1
Dinethoate EPA-1 & 2
Bentazon EPA-1
Propylene Glycol EPA-1
GLC-TCD-IS
Vacor (trade name) EPA-1 & 2
Chlorophacinone EPA-1
Chlorophacinone EPA-1
Dimethoate EPA-1 & 2
Vacor (trade name) EPA-1 & 2
Dimethoate EPA-1 & 2
Dimethoate EPA-1 & 2
Chlorophacinone EPA-1
Linuron EPA-3
Crotoxyphos EPA-1
Methidathion EPA-1
Mercaptobenzothiazole EPA-1 & 2
IR
GLC-FID-IS
Trichlorfon EPA-1 & 2
GLC-TCD-IS
-------
Trimetion
Tugon
Ultracide
Vacor
Vacor (trade name) EPA-1 (tentative)
Vacor (trade name) EPA-2 (tentative)
Diraethoate EPA-1 & 2
Trichlorfon EPA-1 & 2
Methidathion EPA-1
Vacor (trade name) EPA-1 & 2
UV
HPLC
Zetax (zinc salt)
Mercaptobenzothiazole EPA-1 & 2
-------
May 1977
Antimycin A
(tentative)
EPA-1
Determination of Antimycin A
by Ultraviolet Spectroscopy
Antimycin A is a registered piscicide, consisting of a mixture
of antimycin A- and antimycin A- which have the following chemical
names and structures:
antimycin A.; Butanoic acid, 3-methyl —3— ||3-(forraylamino)-2
-hydroxybenzoyljamlnoi-8-hexyl-2,6-dimethyl-4,9-dioxo-l,5
-dioxonan-7-yl ester
antimycin A.; 3-methylbutanoic acid 8-butyl-3- Jfo-^ormylanino)
t 1 ^
-2-hydroxybenzoyll amino -2,6-dimethy1-4,9-dioxo-l,5-dioxonan
-7-yl ester
0—C-CH2-CH
0
H-N-CHO
antimycin AI - R - hexyl -
antimycin A - R - butyl ~
-------
Antimycin A EPA-1
(tentative)
Molecular formula: A. « C?gH,-N_0
A3 ' C26E36N2°9
Molecular weight: A, - 548.62
A3 - 520.56
Melting point: ^ - 149-150°C
A3 - 170.5-171.5°C
Physical state, color, and odor: white solid
Solubility: practically insoluble in water; soluble in acetone, alcohol,
chloroform, benzene; A- is very slightly soluble in benzene
and carbon tetrachloride, but A. is freely soluble
Stability:
Other names: Fintrol
Reagents;
1. Antimycin A standard of known % purity
2. Methanol, spectro or pesticide grade
Equipment;
1. Ultraviolet spectrometer, double beam ratio recording with
matched 1 cm silica cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
-------
Antimycin A EPA-1
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.05 g antlraycin A standard Into a 100 ml volumetric
flask, dissolve In, and make to voltme with methanol. Plpet 2 ml
Into a second 100 ml volumetric flask, make to volume with methanol,
and mix thoroughly, (final cone 10 ug antimycln A/ml)
Preparation of Sample;
For liquids - weigh a portion of sample equivalent to 0.05 gram
antlmycin A into a small beaker. Heat on steam bath under a gentle
stream of air to remove solvents present in the formulation
(usually acetone). Cool, dissolve in 40-50 ml methanol,and
quantitatively transfer to 100 ml volumetric flask, rinsing beaker
several times with methanol. Make to volume with methanol and mix
thoroughly. Pipet 2 ml to a second 100 ml volumetric flask, make
to volume with methanol, and mix thoroughly, (final cone 10 jig
antimycin A/ml)
For wettable powders - weigh a portion of sample equivalent to
0.05 gram antimycin A into a 250 ml glass-stoppered flask or screw-
cap bottle. Add 100 ml methanol by pipette, close tightly, and
shake for one-half hour. Allow to settle; centrifuge or filter if
necessary, taking precaution to prevent evaporation. Pipette 2 ml
into a 100 ml volumetric flask, make to volume with raethanol, and
mix thoroughly, (final cone 10 ug antimycin A/ml)
UV Determination;
With the UV spectrometer at the optimum quantitative settings,
balance the pen for 0 and 1002 transmission at 223 nm with methanol
in each cell. Scan both sample and standard from 300 nm to 200 nm,
using methanol In the reference cell. Measure the absorbance of
both standard and sample at 223 nm, using the minimum at 280 nm as
baseooint.
-------
Antlmycin A EPA-1
(tentative)
Calculation:
Z antimvcin A • feb8* sample)(cone, std in jig/ml)(Z purity of std)
y (abs. std)(cone, sample in ^ig/ml)
Method submitted by Mark W. Law, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
Any criticisms, suggestions, data, etc. on the use of this method will
be appreciated*
-------
June 1977 Asulam EPA-1 (tentative)
Determination of Asulam by Ultraviolet Spectroscopy
Asulara is the accepted common name for methyl sulfanilylcarbamate,
a registered herbicide having the chemical structure:
S— N— C-0-CH3
^
Molecular formula: CgH.,0N20,S
Molecular weight: 230.0
Melting point: pure material - 143 to 144°C, technical material -
135°C, both with decomposition
Physical state, color, and odor: pure material - odorless white
crystals; technical material - cream to buff powder
Solubility: about 0.5% in water, 34% in acetone, 28% in methanol, less
than 2% in hydrocarbons and chlorinated hydrocarbons;
sodium salt more than 40% in water
Stability: Asulam and its dry salts are very stable for years under
ordinary storage conditions; aqueous solution of the
sodium salt at pH 8.5 is very stable.
Other names: Asulox (May & Baker Ltd); MB 9057; methyl(4-aminobenzene-
sulphonyl) carbamate; asulame (France)
Reagents;
1. Asulam standard of known % purity
2. Ethanol, 95%, ACS (or better grade)
-------
Asulara EPA-1 (tentative)
Equipmenti
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Filtration apparatus
A. Usual laboratory glassware
Procedure:
Preparation of Standard;
Weigh 0.06 gram asulam standard into a 100 ml volumetric
flask. Dissolve in and make to volume with 95% ethanol. Mix
thoroughly, pipette a 10 ml aliquot into a second 100 ml
volumetric flask, make to volume with 95% ethanol, and mix
thoroughly. Pipette a 10 ml aliquot into a third 100 ml
volumetric flask, make to volume with 95% ethanol,and mix
thoroughly, (final cone 6 jig asulam/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.06 gram asulam
into a 250 ml Erlenmeyer flask, add by pipette 100 ml 95% ethanol,
and shake on a mechanical shaker for 30 minutes. Allow to settle,
filter if necessary, and pipette 10 ml into a 100 ml volumetric
flask. Make to volume and mix thoroughly. Pipette 10 ml into
another 100 ml volumetric flask, make to volume with 95% alcohol,
and mix thoroughly, (final cone 6 jig asulam/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
analytical settings for thi; particular instrument being used,
balance the pen for 0 and 3.00% transmission at 263 nm with
-------
3 Asulam EPA-1 (tentative)
95% ethanol in each cell. Scan both the standard and sample from
350 run to 200 run with 95% ethanol in the reference cell. Measure
the absorbance of both standard and sample at 263 nm.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent asulam as follows:
7 « (abs. sample) (cone. std in ug/ml) (% purity s td)
(abs. std)(conc. sample in
Method submitted by David Persch, EPA Product Analysis Laboratory,
Region II, New York, N.Y.
Any comments, criticisms, suggestions, data, etc* concerning the use
of this method will be appreciated.
-------
May 1977
Bentazon EPA-1
(tentative)
Determination of Bentazon and Its Sodium
Salt by Ultraviolet Spectroscopy
Bentazon is the accepted common name for 3-isopropyl-lH-2,l,3
-benzothiadiazin-4(3H)-one 2,2-dioxide, a registered herbicide having
the chemical structure:
CH3
Molecular formula: cioHi2N2°3S
Molecular weight:
Melting point:
240.3
137 to 139 C
Physical state, color, and odor: odorless, white, crystalline solid
Solubility: solubility in grams per 100 grams solvent at 20°C:
acetone 150.7, ethanol 86.1, ethyl acetate 65.0,
ether 61.6, chloroform 18.0, benzene 3.3, water 0.05,
cyclohexane 0.02
Stability: stable under ordinary conditions; non-corrosive; no
degradation under visible light (400-600 nm) but 30Z
degradation under ultraviolet light (200-400 nm)
Other names: Basagran, Poast
-------
Bentazon EPA-1
Ctentative)
Reagents;
1. Bentazon standard of known % purity
2, Sodium pyrophosphate, 1Z aqueous solution - dissolve 5 grams
of Na,P.07.10H20 in water and make to 500 ml.
Equipment;
1. Ultraviolet spectrophotoraeter, double beam ratio recording
with matched 1 cm silica cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure;
Preparation of Standard;
Weigh 0.08 gram bentazon standard into a 100 ml volumetric
flask; dissolve in and make to volume with 1Z sodium pyrophos-
phate solution; mix thoroughly. Pipette 10 ml into a second
100 ml volumetric flask, make to volume with 1% sodium pyrophos-
phate solution, and mix thoroughly. Pipette 5 ml of this solution
into a third 100 ml volumetric flask and make to volume with the
pyrophosphate solution. Mix thoroughly, (final cone A jig
bentazon/ml)
Preparation of Sample;
For wettable powders - weigh a portion of sample equivalent to
0.8 gram bentazon into a 250 ml glass-stoppered flask or screw-cap
bottle. Add 100 ml of 1Z sodium pyrophosphate solution, close
tightly, and shake for 30 minutes. Allow to settle; centrifuge or
filter if necessary. Proceed as in the third paragraph.
-------
Bentazon EPA-1
(tentative)
For liquid formulations - weigh a portion of sample equiv-
alent to 0.8 gram bentazon into a 100 ml volumetric flask, make
to volume with 1Z sodium pyrophosphate solution, and mix
thoroughly. Proceed as below.
Pipette 10 ml of either of the above sample solutions into
a 100 ml volumetric flask, make to volume with 1% sodium pyro-
phosphate, and mix thoroughly. Pipette 10 ml of this solution
into a 100 ml volumetric flask, make to volume with the pyrophos-
phate solution, and mix thoroughly. Pipette 5 ml of this solution
into another 100 ml volumetric flask, make to volume with pyro-
phosphate solution, and mix thoroughly, (final cone 4 jig bentazon/ml)
UV Determination:
Balance pen for 0 and 100% transmission at 223 nm with 1Z
sodium pyrophosphate in each cell. Scan standard and sample from
280 nm to 150 nm with 1% sodium pyrophosphate solution in the
reference cell. Measure absorbance of standard and sample at
223 nm.
Calculation:
From the above absorbances and using the standard and sample
concentrations, calculate the percent bentazon as follows:
. m (aba, sample)(cone, std in ug/ml)(Z purity std)
(aba. std)(cone, sample in ug/ml)
Method submitted by Mark W. Law, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
Any comments, criticisms, suggestions, data, etc. on the use of this
method will be appreciated.
-------
May 1977
Chlorobenzilate EPA-1
(tentative)
Determination of Chlorobenzilate by
Gas Liquid Chromatography
(FID - Internal Standard)
Chlorobenzilate is the common name for ethyl 4,4'-dichlorobenzilate,
a registered acaricide having the chemical structure:
Molecular formula: Ci6Hi
Molecular weight: 325.2
Melting point:
Boiling point:
35-37 C for pure product
156-158°C at 0.07 mm Hg
Physical state, color, and odor: pale yellow solid when pure; the
technical product is a brownish liquid about 90%
pure; characteristic odor
Solubility: practically insoluble in water; soluble in most organic
solvents including petroleum oils
Stability: hydrolyzed by alkali and strong acids
Other names: C-23992, Akar, Folbex, Acaraben (Ciba-Geigy); ethyl 4,4'-
dichlorodiphenylglycollate; Benzilan; Kop-Mite
-------
Chlorobenzilate EPA-1
(tentative)
Reagents;
1. Chlorobenzilate standard of known % purity
2. Heptachlor epoxide standard of known % purity
3. Acetone, pesticide or spectro grade
Note: chloroform may also be used, but acetone is preferred.
4. Internal standard solution - weigh 1.0 gram heptachlor epoxide
into a 100 ml volumetric flask; dissolve in and make to volume
with acetone. Mix well, (cone 10 rag /ml or 10
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column; 4' x 1/4" glass column packed with 3.8% UC-V98 on
80/100 mesh diatoport S (or equivalent column)
3. Precision liquid syringe: 5 or 10 jil
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 230 C
Injection temperature: 260 C
Detector temperature: 260°C
Carrier gas: Helium
Carrier gas pressure: 40 psi - 30 ml/min
Hydrogen pressure: 15 psi - 30 ml/min
Air pressure: 40 psi - 60 ml/min
Chart speed: 0,,25"/min or 15"/hr
-------
Chlorobenzilate EPA-1
(tentative)
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
Procedure:
Preparation of Standard;
Weigh 0.1 gram Chlorobenzilate into a 100 ml volumetric flask,
add 10 ml internal standard solution by pipette, make to volume
with acetone, and mix well, (final cone 1 jig Chlorobenzilate and
1 ug heptachlor epoxide/jul)
Preparation of Sample;
For liquids and emulsifiable concentrates - weigh a portion of
sample equivalent to 0.1 gram Chlorobenzilate into a 100 ml volu-
metric flask, add 10 ml Internal standard solution, make to volume
with acetone, and mix well.
For dusts and wettable powders - weigh a portion of sample
equivalent to 0.2 gram Chlorobenzilate into a 250 ml glass-stoppered
flask or screw-cap bottle; add 100ml acetone by pipette. Close
tightly and shake thoroughly to dissolve and extract the Chloroben-
zilate. Shake mechanically for 10-15 minutes or shake by hand inter-
mittently for 25-30 minutes. Allow to settle; filter or centrifuge
if necessary, taking precaution to avoid loss by evaporation.
Pipette a 50 ml aliquot into a 100 ml volumetric flask, add 10 ml
internal standard by pipette, make to volume with acetone, and mix
thoroughly, (final cone 1 ug Chlorobenzilate and 1 jig heptachlor
epoxide/jil)
-------
Chlorobenzilate EPA-1
(tentative)
Determination:
Inject 3 jil of standard and, if necessary, adjust the instrument
parameters and the volume injected to give a complete separation
within a reasonable time and peak heights of from 1/2 to 3/4 full
scale. The elution order is heptachlor epoxide, then chlorobenzilate.
Repeated injections should give the same peak ratios*
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of chlorobenzilate and hepta-
chlor epoxide from both the standard-internal standard solution and
the sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
I.S. - internal standard - heptachlor epoxide
RF m (wt. I.S.) (2 purity I.S.Hpk. ht. or area chlorobenzilate)
(wt. chlorobenzilate)(Z purity chlorobenzilate)(pk. ht. or area I.S.)
Determine the percent chlorobenzilate for each injection of the
sample-internal standard solution as follows and calculate the average:
% » (wt. I.S.)(Z purity I.S.Kpk. ht. or area chlorobenzilate)
(wt. sample)(pk. ht. or area I.S.)(RF)
Method submitted by George B. Radan, EPA Product Analysis Laboratory,
Region II, New York, N.Y.
Any comments, criticism, suggestions, data, etc. concerning the use of
this method will be appreciated.
-------
May 1977
Chlorophacinone EPA-1
(tentative)
Determination of Chlorophacinone
by Ultraviolet Spectroscopy
Chlorophacinone is the coonon name for 2- l(p-chlorophenyl)
phenylacetyl -1,3-lndandione, a registered rodenticide having the
chemical structure:
Molecular formula:
Molecular weight: 374.6
Melting point:
140°C
Physical state, color, and odor: odorless, white crystalline solid
Solubility: sparingly soluble in water; soluble in organic solvents
such as acetone, ethanol, ethyl acetate
Stability: stable and resistant to weathering; non-corrosive;
compatible with cereals, fruits, roots, and other
rodenticide baits; oxidized in bait formulations
Other names: Rozol (Chempar Chemical Co.); Caid, Liphadione, and
Raviac (Lipha SA); Drat (May & Baker Ltd.); Quick (Rhone-
Poulenc); Partox; 2-(2-p-chlorophenyl-2-phenylacetyl)
indane-1,3-dione
-------
Chlorophacinone EPA-1
(tentative)
Reagents:
1. Chlorophacinone standard of known Z purity
2. Sodium pyrophosphate, 1Z solution - dissolve 5 grams
Na.P.O-.lOH.O in water and make to 500 ml.
427 2
3. Sodium pyrophosphate/dioxane mixture - add 10 ml dioxane
to 200 ml of the 1Z sodium pyrophosphate solution.
(Dioxane is added to help dissolve the Chlorophacinone.)
4. Ether-hexane mixture - extract 200 ml n-hexane (bp 60°-68°C)
with three 20 ml portions of 1Z pyrophosphate solution and
add 50 ml ethyl ether making a 20Z ether-80Z hexane mixture.
5. Hydrochloric acid, 2.5N solution - 20.6 ml hydrochloric
acid diluted to 100 ml
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm quartz cells
2. Centrifuge with 50 ml glass-stoppered tubes
3. Mechanical shaker
4. Usual laboratory glassware
Procedure:
Preparation of Standard;
Weigh 0.04 gram Chlorophacinone standard into a 100 ml volu-
metric flask, dissolve in and make to volume with dioxane/sodium
pyrophosphate mixture, mix thoroughly. Pipette 10 ml into a second
100 ml volumetric flask, make to volume with 1Z pyrophosphate
solution, and mix thoroughly. Pipette 10 ml of this solution into
a 50 ml volumetric flask, make to volume with the 1Z pyrophosphate
solution, and mix thoroughly, (final cone 8 pg/ml)
-------
Chlorophacinone EPA-1
(tentative)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.0008 gram chloro-
phacinone (0.4 gram for a 0.2% product) into a 250 ml glass-
stoppered Erlenmeyer flask, add by pipette 100 ml dioxane/
pyrophosphate mixture, close tightly, and shake on a mechanical
shaker for one hour. Transfer 30-35 ml to a glass-stoppered
centrifuge tube and centrifuge for 5 minutes. Pipette 25 ml of
clear solution into a 125 ml glass-stoppered Erlenmeyer flask,
add 5 ml 2.5N hydrochloric acid and 50 ml ether-hexane solution,
stopper tightly, and shake for 5 minutes.
Pipette 20 ml of the ether layer into another centrifuge
tube and add by pipette 10 ml 1% pyrophosphate solution. Shake
for 2 minutes and remove the ether layer — this is conveniently
done by using a tube drawn into a fine tip and connected to a
water aspirator. If the aqueous phase is not clear, centrifuge
for a few minutes with the top off to remove any traces of the
ether-hexane phase, (final cone 8 jig chlorophaclnone/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
settings, balance the pen for 0 and 100% transmission at 285 nm
with 1% pyrophosphate solution in each cell. Scan both standard
and sample from 360 nm to 240 nm, using 1% pyrophosphate solution
in the reference cell. Measure the absorbance of both standard
and sample at 285 nm.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent chlorophacinone as follows:
% » (abs. sample) (vt^ 8td,)(Z purity, std.)
(abs. std.)(wt. sample!
Method submitted by Mark W. Law, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
-------
April 1977
Crotoxyphos EPA-1
(tentative)
Determination of Crotoxyphos
by Gas Liquid Chromatography
FID - Internal Standard
Crotoxyphos is the common name (ISO and BSI) for dimethyl
phosphate of alpha-methylbenzyl 3-hydroxy-cis-crotonate, a
registered insecticide having the chemical structure:
CH3-0 J r „
>P"~~"0*~~C=rCH~~~C' 0 C
y
H
Molecular formula: C.-IL-O,?
Molecular weight: 314.3
Boiling point: 135°C at 0.03 mm Hg (technical grade)
Physical state, odor, and color: light straw-colored liquid
with a mild ester odor
Solubility: 0.1% in water at room temperature; slightly soluble in
kerosene and saturated hydrocarbons; soluble in acetone,
chloroform, ethanol, isopropanol, and highly chlorinated
hydrocarbons; miscible with xylene
Stability: stable in hydrocarbon solvents, but hydrolyzed by water;
decomposes in acidic or basic solution; slightly corrosive
to mild steel, copper, lead, zinc, and tin; non-corrosive
to stainless steel 316, monel, aluminum 3003; will not
attack rigid PVC, fiber glass, reinforced polyester or
the usual lacquers used for lining drums; formulations
made with common pesticide carriers are not stable for
-------
Crotoxyphos EPA-1
(tentative)
Other names: Ciodrin (Shell), SD 4294, dimethyl-2-(alpha-methyl-
benzocarbonyl)-l-methyl vinyl phosphate(E), 1-methyl-
benzyl 3-(dimethoxyphosphinyloxy)-cis-crotonate,
dimethyl cis-l-«ethyl-2-(l-phenylethoxycarbonyl)
vinyl phosphate
NoteI This method was developed and is used by the EPA Beltsville
Chemistry Laboratory. The Kentucky Division of Regulatory
Services uses a very similar method—data is given for
their method following this EPA method.
Reagents;
1. Crotoxyphos standard of known % purity
2. Dipentyl phthalate standard of known % purity
3. Acetone, pesticide or spectre grade
4. Internal Standard solution - weigh 1.0 gram dipentyl
phthalate standard into a 50 ml volumetric flask; dissolve
in and make to volume with acetone, (cone 20 mg dipentyl
phthalate/ml)
Equipment;
1. Gas chromatograph with flame ionizatlon detector (FID)
2. Column: 6* x 2 mm ID glass column packed with 5% SE-30 on
Chromosorb W DMCS AW (or equivalent column)
3. Precision liquid syringe: 5 or 10 jil
4. Usual laboratory glassware
-------
Crotoxyphos EPA-1
(tentative)
Operating Conditions for FID;
Column temperature: 205 C
Injection temperature: 250 C
Detector temperature: 250°C
Carrier gas: Nitrogen or helium
Carrier gas pressure: 30 psl (adjusted for specific GC)
Hydrogen pressure: 20 psi (adjusted for specific GC)
Air pressure: 30 psi (adjusted for specific GC)
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.1 gram crotoxyphos standard into a 25 ml volumetric
flask. Add by pipette 5 ml of the internal standard solution,
make to volume with acetone, and shake thoroughly, (final cone
4 mg crotoxyphos and 4 mg dipentyl phthalate/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.1 gram crotoxyphos
into a 25 ml volumetric flask. Add by pipette 5 ml of the
internal standard solution. Dissolve and make to volume with
acetone, (final cone 4 mg crotoxyphos and 4 mg dipentyl
phthalate/ml)
-------
Crotoxyphos EPA-1
(tentative)
Determination:
Inject 5 jil of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is crotoxyphos, then
dipentyl phthalate.
Proceed with the determination, making at least three
injections each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of crotoxyphos and dipentyl
phthalate from both the standard-internal standard solution and
the sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
I.S. « internal standard » dipentyl phthalate
..p m (wt. I.S.)(% purity I.S.)(pk. ht. or area crotoxyphos)
(wt. crotoxyphos)(% purity crotoxyphos)(pk. ht. or area I.S.)
Determine the percent crotoxyphos for each injection of the
sample-Internal standard solution as follows and calculate the
average:
. m (wt. I.S.)(Z purity I.S.)(pk. ht. or area crotoxyphos)
(wt. sample)(pk. ht. or area I.S.)(RF)
Method submitted by Mark W. Law, EPA Beltsville Chemistry Laboratory,
Beltsville, Maryland.
-------
Crotoxyphos EPA-1
(tentative)
A very similar method, differing as noted below, was submitted by
the Division of Regulatory Services, Kentucky Agricultural Exper-
iment Station, University of Kentucky, Lexington, Kentucky 40506.
Column: 6* x 2 mm ID glass column packed with 3% OV-1 on 60/80 mesh
Gas Chrom Q
Internal Standard: dibutyl phthalate
Cone, of crotoxyphos: 10 mg/ml in both standard and sample solutions
Cone, of internal standard: 2.5 mg/ml in both standard and sample
solutions
Volume injected: 1.5 ul
Column temperature: 230°C
Injection temperature: 300°C
Detector temperature: 300°C
Carrier gas: Nitrogen - 60 psi or 13.3 ml/rain
Hydrogen: 34 psi or 30 ml/min
Air: 28 psi or 300 ml/min
Instrument: Perkin Elmer 900 or Varian 2700
The above conditions should be adjusted to give optimum results
with the particular GC used.
Note' These methods have been designated as tentative. Any comments,
criticisms, suggestions, data, etc. concerning these methods
will be appreciated, especially as related to analysis of
different crotoxyphos formulations.
-------
June 1977 Dimethoate EPA-1
(tentative)
Determination of Dimethoate by
Gas Liquid Chromatography
(TCD - Internal Standard)
Dimethoate is the accepted common name for 0,0-dimethyl S-£(methyl-
carbamoyl) methyl] phosphorodithioate, a registered insecticide having
the chemical structure:
CH3-0 S o ^
—S—CH2— C— N—CH3
Molecular formula: C5H.2NO-PS2
Molecular weight: 229.1
Melting point: pure compound - 51 to 52°C; technical - 43 to 50°C
Physical state, color, and odor: pure compound forms colorless crystals
and has a camphor-like odor; technical compound has
a mercaptan odor
Solubility: 2-3% in water; most soluble in polar solvents such as
alcohols and ketones (acetone and cyclohexanone); lower
solubility in non-polar solvents such as xylene and hexane
Stability: stable in aqueous solutions; hydrolyzed by aqueous alkali;
heating converts dimethoate to the -SCH- isomer; incompat-
ible with alkaline pesticides
Other names: E.I. 12,880, Cygon, Dimetate (American Cyanamid); L395,
Fostion MM, Rogor (Montecatini); Roxion (Cela); Perfekthion
(BASF); Daphene, De-Fend, Dimethogen, Rebelate, Trimetion
-------
Dlmethoate EPA-1
(tentative)
Reagents;
1. Dlmethoate standard of known % purity
2. Heptachlor standard of known % purity
3. Chloroform, pesticide or spectro grade
4. Internal Standard solution - weigh 1 gram heptachlor standard
into a 25 ml volumetric flask; dissolve in and make to volume
with chloroform; mix well, (cone 40 mg heptachlor/ml)
Equipment;
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 4' x 1/4" I.D. glass, packed with 5% SE-30 on
Diatoport S or equivalent column (such as 4* x 1/4"
I.D. glass, packed with 5% SP-2100 on 80/100 Chromo-
sorb 750)
3. Precision liquid syringe: 10 ;il
4. Usual laboratory glassware
Operating Conditions for TCD;
Column temperature: 165 C
Injection temperature: 200 C
Detector temperature: 200°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas pressure: 40 psi
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Dimethoate EPA-1
(tentative)
Procedure:
Preparation of Standard;
For use with emulsifiable concentrates and liquid formu-
lations - weigh 0.1 gram dimethoate standard into a 10 ml
volumetric flask; add 5 ml internal standard solution by
pipette, make to volume with chloroform, and mix well.
(final cone 10 rag dimethoate and 20 mg heptachlor/ml)
For use with dusts, granules, and wettable powders -
weigh 0.1 gram dimethoate standard into a small glass-
stoppered flask or screw-cap bottle, add by pipette 5 ml
of internal standard solution and 5 ml chloroform, close
tightly and shake well to dissolve the dimethoate. (final
cone 10 mg dimethoate and 20 mg heptachlor/ml)
Preparation of Sample;
For emulsifiable concentrates and liquid formulations -
weigh a portion of sample equivalent to 0.1 gram dimethoate
into a 10 ml volumetric flask; add 5 ml of internal standard
solution by pipette, make to volume with chloroform, and mix
well, (final cone 10 rag dimethoate and 20 mg heptachlor/ml)
For dusts, granules, and wettable powders - weigh a
portion of sample equivalent to 0.1 gram dimethoate into a
small glass-stoppered flask or screw-cap bottle; add by
pipette 5 ml internal standard and 5 ml of chloroform, close
tightly, and shake on a mechanical shaker for 10-15 minutes or
shake by hand intermittently for 25-30 minutes. (final cone
10 mg dimethoate and 20 mg heptachlor/ml)
-------
Dlmethoate EPA-1
(tentative)
Determination:
Inject 2 jil of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a
complete separation within a reasonable time and peak heights
of from 1/2 to 3/4 full scale. The elution order is dimethoate,
then heptachlor.
Proceed with the determination, making at least three
injections each of standard and sample solutions in random
order.
Calculation:
Measure the peak heights or areas of dimethoate and
heptachlor from both the standard-internal standard solution
and the sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(wt. heptachlor)(% purity heptachlor)(pk. ht. or areti dimethoate)
(wt. dimethoate)(% purity dimethoate)(pk. ht. or area heptachlor)
Determine the percent dimethoate for each injection of the
sample-internal standard solution as follows and calculate the
average:
(wt. heptachlor)(% purity heptachlor)(pk. ht. or area dimethoate)
(wt. sample)(pk. ht. or area heptachlor)(RF)
Method submitted by Stelios Gerazounis, EPA Product Analysis Lab,
Region II, New York, N.Y. (experimental method May 1970)
Any comments, criticisms, suggestions, data, etc. concerning this
method or its use will be appreciated.
-------
June 1977 Dimethoate EPA-2
(tentative)
Determination of Dimethoate by
Gas Liquid Chromatography
(FID - Internal Standard)
Dimethoate is the accepted common name for 0,0-dimethyl S-L(methyl-
carbamoyl) methyllphosphorodithioate, a registered insecticide having
the chemical structure:
? V f
— CH2—C — N—CH3
Molecular formula: C H._NO-PS-
Molecular weight: 229.1
Melting point: pure compound - 51 to 52°C; technical - 43 to 50°C
Physical state, color, and odor: pure compound forms colorless crystals
and has a camphor-like odor; technical compound has
a mercaptan odor
Solubility: 2-3% in water; most soluble in polar solvents such as
alcohols and ketones (acetone and cyclohexanone); lower
solubility in non-polar solvents such as xylene and hexane
Stability: stable in aqueous solutions; hydrolyzed by aqueous alkali;
heating converts dimethoate to the -SCH_ isomer; incompat-
ible with alkaline pesticides
Other names: E.I. 12,880, Cygon, Dimetate (American Cyanamid); L395,
Fostion MM, Rogor (Montecatini); Roxion (Cela); Perfekthion
(BASF); Daphene, De-Fend, Dimethogen, Rebelate, Trlmetion
-------
Dlmethoate EPA-2
(tentative)
Reagents;
1. Dimethoate standard of known % purity
2. Dibutyl phthalate standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 1.5 gram dibutyl phthalate
into a 200 ml volumetric flask; dissolve in and make to volume
with acetone; mix well. (cone 7.5 mg dibutyl phthalate/ml)
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: A1 x 2 mm glass column packed with 5% SE-30 on
80/100 Chromosorb W HP (or equivalent column)
3. Precision liquid syringe: 5 or 10 pi
4. Mechanical shaker
*
5. Usual laboratory glassware
* Virginia laboratories place their weighed samples in
25 mm x 200 mm screw-top culture tubes, add solvent by
pipette, and seal tightly with teflon-faced rubber-lined
screw caps.
These tubes are rotated end over end at about 40-50
RPM on a standard Patterson-Kelley twin shell blender that
has been modified by replacing the blending shell with a
box to hold a 24-tube rubber-covered rack.
Operating Conditions for FID;
Column temperature: 170 *C
Injection temperature: 220°C
Detector temperature: 220 C
-------
3 Dimethoate EPA-2
(tentative)
Carrier gas: Nitrogen
Carrier gas pressure: 60 psi (adjusted as necessary)
Hydrogen pressure: 20 psi (adjusted as necessary)
Air pressure: 30 psi (adjusted as necessary)
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.1 gram dimethoate standard into a 50 ml volumetric
flask, add 10 ml internal standard solution by pipette, make
to volume with acetone, and shake to dissolve, (final cone
2 mg dimethoate and 1.5 mg dibutyl phthalate/ml)
Preparation of Sample;
For emulsifiable concentrates and liquid formulations -
weigh a portion of sample equivalent to 0.1 gram dimethoate
into a 50 ml volumetric flask; add 10 ml internal standard
solution, make to volume with acetone, and mix well, (final
cone 2 mg dimethoate and 1.5 mg dibutyl phthalate/ml)
For dusts, granules, and wettable powders - weigh a
portion of sample equivalent to 0.1 gram dimethoate into a
glass-stoppered flask or screw-cap bottle; add by pipette
10 ml internal standard solution and 40 ml acetone, close
tightly, and shake on a mechanical shaker for 10-15 minutes or
shake by hand intermittently for 25-30 minutes, (final cone
2 mg dimethoate and 1.5 mg dibutyl phthalate/ml)
-------
Dimethoate EPA-2
(tentative)
Determination:
Inject 1-2 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a com-
plete separation within a reasonable time and peak heights of
from 1/2 to 3/4 full scale. The elution order is dimethoate,
then dibutyl phthalate.
Proceed with the determination, making at least three
injections each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of dimethoate and dibutyl
phthalate from both the standard-internal standard solution and
the sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
DBF - dibutyl phthalate
RF = (wt. DBP)(% purity DBF)(pk. ht. or area dimethoate)
(wt. dimethoate)(% purity dimethoate)(pk. ht. or area DBF)
Determine the percent dimethoate for each injection of the
sample-internal standard solution as follows and calculate the
average:
"/ = (wt. DI?P)(% purity DBF) (pk. ht. or area dimethoate)
(wt. sample)(pk. ht. or area DBF)(RF)
This method was submitted by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond, Va. 23219.
This method has been designated as tentative since it is a Va. Exp. method
and because some of the data has been suggested by EPA's Beltsville Chem-
istry Lab. Any comments, criticism, suggestion, data, etc. concerning
this method will be appreciated.
-------
June 1977 Ethylenethiourea EPA-1
(tentative)
Determination of Ethylenethiourea in
Ethylenebisdithiocarbaraate Fungicides
by Gas Liquid Chromatography (FID and TCD)
Ethylenethiourea (ETU) is a degradation product of ethylene-
bisdithiocarbamates (EBDC). It may be formed during manufacture,
storage, or use of EBDC fungicide formulations.
The following methods are not residue methods in the sense
of measuring ETU in raw crops, processed foods, soil, or water.
However, these methods will show the presence of ETU in formulated
EBDC fungicide products.
Chemically, ETU is 2-imidazolidinethione and has the structure:
V
•N-H
Molecular formula: C0H,N0S
JO/
Molecular weight: 102.17
Melting point: 203-204°C (technical: 199-204°C)
Physical state, color, and odor: white to pale green crystals,
faint amine odor
Solubility: solubility in 100 ml water - 2 g at 30°, 9 g at 60°,
44 g at 90 ; moderately soluble at room temperature in
methanol, ethanol, ethylene glycol, pyridine, acetic
acid, and naphtha; insoluble in acetone, ether, chloro-
form, benzene, and ligroin
-------
Ethylenethiourea EPA-1
(tentative)
Stability: dry crystals are stable under usual laboratory storage
conditions; solutions in water or methanol are reason-
ably stable for 6 months to 1 year; somewhat affected
by ultraviolet light; reported to be unstable as residue
on crops or upon cooking
Other names: ETU
The method includes a procedure for using either a flame
ionization detector (FID) or a thermal conductivity detector (TCD).
The FID with its higher sensitivity is more useful for determining
small amounts of ETU. However, since the TCD is non-destructive,
it permits collection of ETU and other eluted components for
Infrared or Mass Spectrometry confirmation.
Reagepts;
1. ETU standard of known % purity
2. Methanol, pesticide or spectro grade — add about 10 grams
anhydrous sodium sulfate per 100 ml to minimize the effect
on any water present in the alcohol.
3. Sodium sulfate, anhydrous, granular
Equipment;
1. Gas chromatograph with flame ionization detector and/or
thermal conductivity detector
2. TCD column: 3' x 1/4" OD stainless steel packed with 2%
Carbowax 20M on Chromosorb W AW DMCS (or
equivalent column — see note below)
3. FID column: 6* x 2 mm ID glass packed with 2% SP-1000
on Chromosorb 750 (or equivalent column —
see note below)
-------
3 Ethylenethiourea EPA-1
(tentative)
4. Precision liquid syringe: 50 ;il (TCD) and/or 10 jil (FID)
5. Screw-cap test tubes: 16 mm x 150 mm
6. Centrifuge (for above tubes)
Note! Carbowax 20M and SP-1000 are equally effective (very
similar McReynolds Constants) in the determination of
ETU. OV-225 and XE-60 have also been used. OV-225
produces less tailing with methanol and ETU.
Supelcoport, Chromosorb W AW DMCS, Chromosorb W HP,
and Chromosorb 750 all have been used and are satis-
factory solid supports.
Any combination of these or other stationary phases
and solid supports may be used if the ETU peak is
reasonably symmetrical and well separated from the
solvent tail.
Determination using Thermal Conductivity Detector;
Operating Conditions;
Column temperature: 220 C
Injection temperature: 270 C
Detector temperature: 250 C
Filament current: 250 ma
Carrier gas: Helium
Carrier gas pressure: 40 psi (adjusted for specific GC)
Carrier gas flow: 100 ml/min (adjusted for specific GC)
Operating conditions for filament current, column temperature,
or gas flow should be adjusted by the analyst to obtain optimum
response and reproducibility.
-------
Ethylenethlourea EPA-1
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.2 gram ETU standard into a 100 ml volumetric
flask; dissolve in and make to volume with methanol (cone
2 >ig ETU/nl)
Preparation of Sample;
Weigh 1 gram of the ethylenebisdithiocarbamate fungi-
cide sample into a 16 mm x 150 nun screw-cap test tube and
add approximately 1 gram anhydrous sodium sulfate. Add
5 ml methanol by pipette, close tightly, and shake inter-
mittently over a period of one hour. Centrifuge until a
clear liquid layer is obtained. (The sides of the tube may
be washed down with the clear layer and the tube re-centri-
fuged for final clarification of the liquid layer.)
One gram of sample in 5 ml methanol gives a sample
concentration of 200 ug/ul which is equivalent to a concen-
tration of 2 ug ETU/ul at the 1% ETU level or of 0.2 jig
ETU/jil at the 0.1% ETU level. For higher concentrations,
10 or 15 ml of methanol may be used instead of only 5 ml
with 1 gram of sample, or a smaller sample size may be used.
Determination;
Using a precision liquid syringe, alternately inject
three 5-80 jil portions each of standard and sample solutions
depending on the amount needed to give a measurable size
peak. Measure the peak area for each peak and calculate the
average for both standard and sample.
Adjustments in attenuation or amount injected may have
to be made to give convenient size peaks, especially with
samples containing very small amounts of ETU. The amount
injected is limited by the size of the methanol peak.
-------
Ethylenethiourea EPA-1
(tentative)
Using a Beckman GC-2A gas chromatograph (no longer
used) on which the injection port, column, and detector
were all at the same temperature, the standardization
curve using peak area vs. jil of ETU standard was linear
from 10 to 160 ;ig. Other gas chromatographs where the
injection port and detector can be at a higher temperature
than the column will give a better response, as will a
more sensitive detector. Under these conditions the
linearity may be extended down to 5 ug or less.
Calculation:
From the average peak area calculate the percent ETU
as follows:
'/ «> (pk. area sample) (wt. std injected)(% purity of std)
(pk. area standard)(wt. sample injected)
Collection of ETU for Infrared and Mass Spectrometry;
The exhaust port of the thermal conductivity detector
is modified by attaching a piece of 1/8" stainless steel
tubing covered with Teflon tubing. The exact length depends
on the configuration needed for the particular detector.
The Teflon tubing should extend about one inch past the end
of the stainless steel tubing to allow insertion of about
1" of a 6-inch piece of 3 mm glass tubing. The entire
length of Teflon tubing is wound with a flexible heating
tape attached to a variac. It is convenient to include a
thermometer (preferably dial type with stainless steel shaft
also covered by Teflon) placed along the side of the heated
tube. The variac should be set so that the temperature is
the same as the detector or about 10 C higher.
-------
Ethylenethiourea EPA-1
(tentative)
The ETU is condensed in the glass tube, usually within
a one-inch length of the air-cooled tube just outside of
the heated portion. No special cooling is needed.
For IR identification, the ETU is washed from the
glass tube with 4 or 5 50 jil portions of methanol into a
small (1 or 2 ml) Mini-Vial (small vial with cone-shaped
interior) containing a few milligrams of potassium bromide.
The KBr and methanol solution is stirred with a small glass
rod drawn into a long fine tip. Gentle heat is applied
until the methanol evaporates completely. The KBr (with
ETU) is then placed into a micro-pellet press, formed into
a disk, and scanned on an IR spectrophotometer from 4000 cm
to 250 cm" (2.5 p to 40 p). A similar pellet is made from
a portion of ETU standard solution and the IR scans compared.
For mass spectrometer identification, the ETU is washed
from the glass tube into a 1 ml Kuderna-Danish concentrator
tube, evaporated to a convenient workable volume, and injected
into a GC-MS.
Determination using Flame lonization Detector;
Operating Conditions;
Column temperature: 180 C
Injection temperature: 270 C
Detector temperature: 270 C
Carrier gas: Helium
Operating conditions for column temperature, carrier gas flow,
or hydrogen/air flow rates should be adjusted by the analyst to
obtain optimum response and reproducibility.
-------
Ethylenethiourea EPA-1
(tentative)
Procedure:
preparation of Standard;
Weigh 0.2 gram ETU standard into a 100 ml volumetric
flask; dissolve and make to volume with methanol (final
cone 2 mg ETU/pl)
Preparation of Sample;
Weigh 1 gram of the ethylenebisdithiocarbamate
fungicide sample into a 16 x 150 mm screw-cap test tube
and add approximately 1 gram of anhydrous sodium sulfate.
Add 5 ml raethanol by pipette, close tightly, and shake
intermittently over a period of one hour. Centrifuge until
a clear liquid layer is obtained. (The sides of the tube
may be washed down with the clear layer and the tube re-
centrifuged for final clarification of the liquid layer.)
One gram of sample in 5 ml methanol gives a sample
concentration of 200 jig/jil which is equivalent to a concen-
tration of 2 ug ETU/pi at the 1% ETU level or of 0.2 jig
ETU/jil at the 0.1% ETU level. For higher concentrations of
ETU, 10 or 15 ml of methanol may be used instead of only
5 ml with 1 gram of sample, or a smaller sample size may be
used.
Determination:
Using a precision liquid syringe, alternately inject
three 2-3 pi portions each of standard and sample solutions.
Measure the peak height or peak area for each peak and cal-
culate the average for both standard and sample.
Adjustments in attenuation or amount injected may have
to be made to give convenient size peaks.
-------
Ethylenethiourea EPA-1
(tentative)
Calculation;
From the average peak height or peak area calculate
the percent ETU as follows:
y m (pk« ht. or area sample)(wt. std injected) (%_ jgurity of std)
(pk. ht. or area standard)(wt. sample injected)
This method (including both TCD and FID) has been used by EPA
Beltsville Chemistry Lab for the last six years. The collection
technique has been developed by Jack Looker.
The method is designated as tentative since several different
columns and gas chromatographs have been used throughout these six
years.
Any comments, criticism, suggestions, or data concerning the
use of this method will be greatly appreciated.
-------
April 1977
Llnuron EPA-3
(tentative)
Determination of Llnuron
by Ultraviolet Spectroscopy
Linuron is the common name for 3-(3,4-dichlorophenyl)-l-methoxy
•1-methylurea, a registered herbicide having the chemical structure:
CH3 0 H
I II I
CH3—0—N C—N
Molecular formula: C9H10C12N2°2
Molecular weight: 249.1
Melting point: 93 to 94°C
Physical state, color, and odor: odorless, white, crystalline solid
Solubility: 75 ppm in water at 25°C; slightly soluble in aliphatic
hydrocarbons, moderately soluble in ethanol and common
aromatic solvents, soluble in acetone
Stability: stable at its m.p. and in solution; slowly decomposed by
acids and bases in moist soil; non-corrosive
Other names: Lorox (DuPont), Afalon, Sarclex, HOE 2810
Reagents;
1. Linuron standard of known % purity
2. Methanol, pesticide or spectro grade
-------
Linuron EPA-3
(tentative)
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of Standard;
Weigh 0.1 gram linuron standard into a 100 ml volumetric
flask. Dissolve in and mike to volume with raethanol; mix
thoroughly. Pipette a 10 ml aliquot into a second 100 ml
volumetric flask, make to volume with methanol, and mix thor-
oughly. Pipette a 5 ml aliquot into a third 100 ml volumetric
flask, make to volume with methanol, and again mix thoroughly.
(final cone 5 ug linuron/ral)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.1 gram of linuron
into a 250 ml Erlenmeyer glass-stoppered flask. Add 100 ml
methanol by pipette and shake on a mechanical shaker for 30
minutes. Allow to settle; filter or centrifuge if necessary,
taking precautions to avoid evaporation. Pipette 10 ml into a
100 ml volumetric flask, make to volume with methanol, and mix
thoroughly. Pipette 5 ml into another 100 ml volumetric flask,
make to volume with methane], and again mix thoroughly. (final
cone 5 ug linuron/ml)
-------
Linuron EPA-3
(tentative)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 246 nm with
methanol in each cell. Scan both the standard and sample from
350 nm to 200 nm with methanol in the reference cell. Measure
the absorbance of both standard and sample at 246 nm.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent linuron as follows:
9 m (abs« sample)(cone, std in ug/ml)(% purity std)
(abs. std)(cone, sample in ug/ml)
Beer's Law is followed from 1 to 10 jig/ml.
Method submitted by David Persch, EPA Product Analysis Lab, Region II,
New York, N. Y.
-------
April 1977
Mercaptobenzothiazole EPA-1
(tentative)
Determination of 2--Mercaptobenzothiazole
(Sodium Salt) by Ultraviolet Spectroscopy
2-mercaptobenzothiazole is a registered fungicide having the
chemical structure:
Molecular formula: CjHJflS-
Molecular weight: 167.25
Melting point: 180.2 to 181.7°C (technical product - 170° to 175°C)
Physical state, color, and odor: pale yellow, monoclinic needles or
leaflets; disagreeable odor
Solubility: practically insoluble in water; solubility at 25°C - 2%
in alcohol, 1% in ether, 10% in acetone, 1% in benzene,
•^0.2% in carbon tetrachloride,<0.5% In naphtha; moderately
soluble in glacial acetic acid; soluble in alkalies and
alkali carbonate solutions
Stability:
Other names: 2-benzothiazolethiol, MBT, Captax, Dermacid, Mertax,
Thiotax, Nuodex 84 (sodium salt), Bantex and Zetax
(zinc salts), Niacides (mixtures with carbamate fungicides)
-------
Mercaptobenzothlazole EPA-1
(tentative)
Reagents;
1. 2-mercaptobenzothiazole standard of known % purity
2. Sodium hydroxide -0.5% solution in water
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Usual laboratory glassware
Procedure;
Preparation of Standard;
Weigh 0.09 gram 2-mercaptobenzothiazole standard into a 500 ml
volumetric flask, dissolve in and make to volume with 0.5% sodium
hydroxide solution, and mix thoroughly. Pipette a 25 ml aliquot
into a 100 ml volumetric flask, make to volume with 0.5% sodium
hydroxide solution, and mix thoroughly. Pipette 10 ml of this
solution into another 100 ml volumetric flask, again make to
volume with 0.5% sodium hydroxide solution, and mix thoroughly.
(final cone 4.5 ug 2-mercaptobenzothiazole/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.09 gram 2-mercapto-
benzothiazole into a 500 ml volumetric flask, dissolve in and make
to volume with 0.5% sodium hydroxide solution, and mix thoroughly.
Pipette a 25 ml aliquot into a 100 ml volumetric flask, make to
volume with 0.5% sodium hydroxide solution, and mix thoroughly.
Pipette 10 ml of this solution into another 100 ml volumetric
flask, again make to volume with 0.5% sodium hydroxide solution,
and mix thoroughly, (final cone 4.5 ug 2-mercaptobenzothiazole/ml)
-------
Mercaptobenzothiazole EPA-1
(tentative)
Note: Samples in paste form have been successfully extracted by
one hour shaking.
UV Determination;
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100Z transmission at 308 nm with 0.5%
sodium hydroxide solution in each cell. Scan both the standard
and sample from 360 nm to 250 nm with 0.52 sodium hydroxide
solution in the reference cell. Measure the absorbance of both
standard and sample at 308 nm.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent 2-mercaptobenzothiazole as
follows:
2 . (abs. sample)concA std in jig/ml) (V purity std)
(abs. std)(cone, sample in jug/ml)
% 2-mercaptobenzothiazole, sodium salt - 1.131 x 2-mercaptobenzothiazole
Method submitted by Edward Yager, EPA Product Analysis Laboratory,
Region II, New York, N.Y.
-------
May 1977
Mercaptobenzothlazole EPA-2
(tentative)
Determination of 2-Mercaptobenzothiazole
(40% Solution of Sodium Salt)
by Potentiometric Titration
2-raercaptobenzothiazole is a registered fungicide having the
chemical structure:
•N
Molecular formula: C_H NS«
Molecular weight: 167.25
Melting point:
180.2 to 181.7°C (technical product - 170° to 175°C)
Physical state, color, and odor: pale yellow, monoclinic needles or
leaflets; disagreeable odor
Solubility: practically insoluble in water; solubility at 25°C - 2%
in alcohol, 1% in ether, 10% in acetone, 1% in benzene,
-------
Mercaptobenzothiazole EPA-2
(tentative)
Reagents;
1. 2-mercaptobenzothiazole, sodium salt standard of known %
purity (see note under Procedure; Calibration of Instrument;)
2. Hydrochloric acid, 0.1 N solution
3. Buffer solutions, pH 4.00 and 7.00
Equipment;
1. pH meter or titrimeter
2. Magnetic stirrer with 1" stirring bar
3. Usual laboratory glassware
Procedure;
Calibration of Instrument;
Calibrate the pH meter or titrimeter at pH 4.00 and pH 7.00.
Note; Each time the instrumentation for this method is
changed (new pH meter, new electrode, repairs, etc.) or at
least once a year, it is desirable to titrate a known
standard to verify proper instrument response and to locate
the inflection point on the titration curve where the 2-mer-
captobenzothiazole will be determined.
Preparation of Sample;
Weigh a portion of sample equivalent to 0.4-0.5 gram 2-
mercaptobenzothiazole (for a 21-26 ml (net) titration) into a
250 ml beaker; add 100 ml distilled water and a 1" magnetic
stirring bar. Place the beaker on a magnetic stirrer, insert
the electrodes, and stir.
-------
Mercaptobenzothiazole EPA-2
(tentative)
Titration:
Titrate with 0.1 N hydrochloric acid solution in small
increments, recording the milliliters added and the corresponding
pH after each addition. On either side of pH 9.5 and pH 5, add
the hydrochloric acid in increasingly smaller amounts, finally
adding the acid drop by drop to obtain a detailed change in the
slope of the titration curve at both inflection points.
Calculation:
Plot the milliliters of hydrochloric acid on the abscissa
and the corresponding pH values on the ordlnate. Draw a smooth
curve through these points. Two inflection points indicate free
sodium hydroxide and the sodium salt of 2-mercaptobenzothiazole.
The first endpoint is taken as the mid-point of the inflection
near pH 9.5 and is the milliliters of 0.1 N hydrochloric acid
used to titrate any free sodium hydroxide according to the reaction:
NaOH + HC1 NaCl + H20
The second endpoint is taken as the mid-point of the inflection
near pH 5 and is the total milliliters of 0.1 N hydrochloric acid
used to titrate any free sodium hydroxide plus the sodium salt of
2-mercaptobenzothiazole (KSNa) according to the reaction:
RSNa + HC1 RSH + NaCl
The percent sodium salt of 2-mercaptobenzothiazole is determined
as follows:
% . (B-A) (N) (0.18924) (100)
(weight of sample)
-------
Mercaptobenzothiazole EPA-2
(.tentative)
where:
B « rals 0.1 N hydrochloric acid at second endpoint
A « mis 0.1 N hydrochloric acid at first endpoint
N • actual N of hydrochloric acid
0.18924 - raeq. wt. of sodium salt of 2-mercaptobenzothiazole
Method submitted by Stelios Gerazounis, EPA Product Analysis
Laboratory, Region II, New York, N.Y.
Any comments, criticisms, suggestions, data, etc. concerning this
method will be appreciated, especially as related to any other
formulation analysis.
-------
June 1977 Methidathion EPA-1
(tentative)
Determination of Methidathion by
Gas Liquid Chroraatography
(FID - Internal Standard)
Methidathion is the accepted common name for 0,0-dlmethyl phos-
phorodithioate S-ester with 4-(mercaptomethyl)-2-methoxy-delta 2-
l,3,4-thiadiazolin-5-one, a registered insecticide and acaricide
having the chemical structure:
CH3-CX S 0=C \—OCH3
II I II
'P—S—CH2—N N
/
CH3-0'
Molecular formula: C,H,,N0O.PS0
O 11 / 4 J
Molecular weight: 302.3
Melting point: 39 to 40°C
Physical state, color, and odor: colorless crystals, characteristic
odor of organophosphates
Solubility: 240 ppm in water at 25°C; readily soluble in acetone,
benzene, methanol
Stability: stable in neutral and weakly acid media but much less
stable in alkali. Compatible with captan, thiram, zineb,
and acaricides.
-------
Methidathion EPA-1
(tentative)
Other names: GS-13005, Supracide, Ultracide (Ciba-Geigy); S-(2,3-
dlhydro-5-methoxy-2-oxo-l,3,4-thiadiazol-3-ylmethyl)
dimethyl phosphorothiolothionate; S-£(5-methoxy-2-
oxo-l,3,4-thiadiazol-3(2H)-yl)methy^ 0,0-dimethyl
phosphorodithioate
Reagents;
1. Methidathion standard of known % purity
2. Aldrin standard of known HHDN content
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh a portion of aldrin
standard equivalent to 0.1 gram HHDN into a 100 ml volu-
metric flask; dissolve in and make to volume with acetone;
mix well. (cone 1 mg HHDN/ml)
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 4 mm glass column packed with 3% OV-1 on
60/80 Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 jil
4. Mechanical shaker
5. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 185°C
Injection temperature: 215 C
Detector temperature: 240 C
Carrier gas: Nitrogen
-------
3 Methidathion EPA-1
(tentative)
Carrier gas pressure: 60 psi (adjusted as necessary)
Hydrogen pressure: 20 psi (adjusted as necessary)
Air pressure: 20 psi (adjusted as necessary)
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.1 gram methidathion standard into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 25 ml of
the internal standard solution and shake to dissolve, (final
cone 4 mg methidathion and 1 mg HHDN/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.1 gram methidathion
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 25 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the methidathion.
For coarse or granular materials, shake mechanically for 10-15
minutes or shake by hand intermittently for 25-30 minutes. (final
cone 4 mg methidathion and 1 mg HHDN/ml)
Determination;
Inject 2-3 jil of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is HHDN, then methidathion.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
-------
Methidathion EPA-1
(tentative)
Calculation;
Measure the peak heights or areas of methidathion and HHDN
from both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
RF
* (wt» HHDN)(pk. ht. or area methidathion^
(wt. methidathion)(% purity methidathion)(pk. ht. or area HHDN)
Determine the percent methidathion for each injection of the
sample-internal standard solution as follows and calculate the
average:
. m (wt. HHDNHpk. ht. or area methidathion)
(wt. sample)(pk. ht. or area HHDN)(RF)
Method submitted by Division of Regulatory Services, Kentucky Agri-
cultural Experiment Station, University of Kentucky, Lexington, Ky. 40506,
Any comments, criticism, suggestions, data, etc. concerning this method
or its use will be appreciated.
-------
April 1977 Monocrotophos EPA-3
(tentative)
Determination of Monocrotophos In Liquid Formulations
by Gas Liquid Chromatography
(FID - Internal Standard)
Monocrotophos is the common name for dimethyl phosphate of
3-hydroxy-N-methyl-cis-crotonamide, a registered insecticide having
the chemical structure:
CH3
P—0—C=C
Molecular formula: C7H..,NOJP
Molecular weight: 223
Melting point: 54 to 55°C (technical material 25 to 30°C)
Physical state, color, and odor: colorless to white crystalline
material with a mild ester odor. The technical
product is a reddish brown semi-solid.
Solubility: miscible with water; soluble in acetone and ethanol;
sparingly soluble in xylene but almost insoluble in
diesel oils and kerosene
Stability: unstable in lower but stable in higher alcohols and
glycols, stable in ketones; hydrolyzes slowly at pH 1
to 7, rapidly above pH 7; corrosive to black iron, drum
steel, brass, SS 304, but does not attack glass, aluminum,
or SS 316; incompatible with alkaline pesticides
-------
2 Monocrotophos EPA-3
(tentative)
Other names: Azodrin (Shell); Nuvacron (Ciba); Monocron; dimethyl-1-
methyl-2-methyl carbamoyl vinyl phosphate; cis-3-
(dimethoxyphosphinyloxy)-N-methylcrotonamide; 0,0-
dimethyl-0-(2-methylcarbamoyl-l-methyl vinyl)-phosphate
Reagents;
1. Monocrotophos standard of known % purity
2. Benzyl benzoate standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 0.45 gram benzyl benzoate
into a 100 ml volumetric flask; dissolve in; and make to volume
with acetone. (cone 4.5 mg benzyl benzoate/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 1/4" glass column packed with 3.8% UC-W98
on 80/100 diatoport S (or equivalent column such as
SP-2100 on Chromosorb 750)
3. Precision liquid syringe: 5 or 10 ul
4. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 175 C
Injection temperature: 200 C (225 may be used)
Detector temperature: 240 C
Carrier gas: Helium (nitrogen may be used)
Carrier gas flow rate: 30 ml/rain - 40 PSI
Hydrogen flow rate: 30 ml/min - 12 PSI
Air flow rate: 55 ml./min - 40 PSI
-------
3 Monocrotophos EPA-3
(tentative)
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.13 gram monocrotophos standard into a 50 ml volumetric
flask. Add (by pipette) 10 ml of the internal standard solution,
swirl to dissolve, and make to volume with acetone. Mix thor-
oughly, (final cone 2.6 mg monocrotophos and 0.9 mg benzyl
benzoate/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.13 gram monocrotophos
into a 50 ml volumetric flask. Add (by pipette) 10 ml of the
internal standard solution; make to volume with acetone and mix
thoroughly. (final cone 2.6 mg monocrotophos and 0.9 mg benzyl
benzoate/ml)
Determination;
Inject 2-4 jil of standard solution and, if necessary, adjust
the instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is monocrotophos, then benzyl
benzoate.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
-------
Monocrotophos EPA-3
(tentative)
Calculation;
Measure the peak heights or areas of monocrotophos and
benzyl benzoate from both the standard-internal standard
solution and the sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
I.S. = internal standard = benzyl benzoate
R_ m (wt. I.S.)(% purity I.S.j^pk. ht. or area monocrotophos)
(wt. monocrotophos)(% purity monocrotophos)(pk. ht. or area I.S.)
Determine the percent monocrotophos for each injection of
the sample-internal standard solution as follows and calculate
the average:
% . (wt» I.S.)(% purity I.S.)(pk. ht. or area monocrotophos)
(wt. sample)(pk. ht. or area I.S.)(RF)
Method submitted by George B. Radan, EPA Product Analysis Laboratory,
Region II, New York, N. Y.
-------
May 1977 Propylene Glycol EPA-1
(tentative)
Determination of Propylene Glycol
by Gas Liquid Chromatography
(TCD - Internal Standard)
Propylene glycol is the common name for 1,2-dihydroxypropane, a
registered disinfectant having the chemical structure:
OH OH
CH3 CH CH2
Molecular formula: C-HgO-
Molecular weight: 76.09
Boiling point: 188.2° at 760 mm Hg; freezes at -59°C
Physical state, color, and odor: colorless, viscous, hygroscopic
liquid; slight odor; slightly acrid taste
Solubility: miscible with water, acetone, alcohols, chloroform, and
many organic solvents; will dissolve many essential oils
but is immiscible with fixed oils
Stability: stable under ordinary conditions; will oxidize at high
temperatures giving such products as propionaldehyde,
lactic acid, pyruvic acid, and acetic acid
f
Other names: 1,2-propanediol; methyl glycol; methylene glycol
Reagents;
1. Propylene glycol standard of known % purity
2. Octyl alcohol standard of known % purity
-------
Propylene Glycol EPA-1
(tentative)
Reagents (cont'd);
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 4 grains octyl alcohol
standard into a 100 ml volumetric flask and make to volume
with acetone; mix well, (cone 40 mg octyl alcohol/ml)
Equipment;
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 41 x 1/4" glass packed with 3% XE-60 on 60/80
Chromosorb G AW DMCS (or equivalent column)
3. Precision liquid syringe: 10 ul
4. Usual laboratory glassware
Operating Conditions for TCDt
Column temperature: 80 C
Injection temperature: 150°C
Detector temperature: 230°C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas flow: 30 ml/min
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.3 gram propylene glycol standard into a 25 ml
volumetric flask, add 10 ml internal standard solution by pipette,
-------
Propylene Glycol EPA-1
(tentative)
Preparation of Standard (cont'd):
and make to volume with acetone; mix well, (final cone 12 jig
propylene glycol and 16 ug octyl alcohol/pi)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.3 gram propylene
glycol Into a 25 ml volumetric flask, add 10 ml Internal standard
solution, and make to volume with acetone; mix well, (final cone
12 jig propylene glycol and 16 fig octyl alcohol/jil)
Determination;
Inject 2-3 ;il of standard-Internal standard solution and, If
necessary, adjust the Instrument parameters and the volume injected
to give a complete separation within a reasonable time and peak
heights from 1/2 to 3/4 full scale. The peak heights of propylene
glycol and octyl alcohol should be nearly the same (definitely
within 25% of each other); if not, concentrations should be
adjusted accordingly. The elution order is propylene glycol, then
octyl alcohol. If the sample contains triethylene glycol, time
should be allowed for this component to be eluted before the next
injection is made.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of propylene glycol and
octyl alcohol from both the standard-internal standard solution
and the sample-internal standard solution.
-------
Propylene Glycol EPA-1
(tentative)
Calculation (cont'd):
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
IS • internal standard - octyl alcohol
PG - propylene glycol
_ (wt. IS)(Z purity IS)j|pk. ht. or area PG)
" (wt. PG)(% purity PG)(pk. ht. or area IS)
Determine the percent propylene glycol for each injection
of sample-internal standard solution as follows and calculate
the average:
y m (wt. IS)(% purity IS)(pk^ht. or area PG)
° * (wt. sample)(pk. ht. or area IS)(RF)
Method submitted by Stelios Gerazounis, EPA Product Analysis Laboratory,
Region II, New York, N.Y.
Any comments, criticisms, suggestions, data, etc. concerning the use
of this method will be appreciated.
-------
April 1977 Trichlorfon EPA-1
(tentative)
Determination of Trichlorfon
by Infrared Spectroscopy
Trichlorfon is the common name (approved by ISO) for dimethyl
(2,2,2-trichloro-l-hydroxyethyl) phosphonate, a registered insecti-
cide having the chemical structure:
0 OH Cl
II I I
P-CH-C—Cl
cH3-cr ci
Molecular formula: C.HgCl-O.P
Molecular weight: 257.5
Melting point: 83-84°C
Physical state, color, and odor: white crystalline solid
Solubility: 15.4 g/100 ml water at 25°C; soluble in benzene, ether,
ethanol, and most chlorinated solvents; slightly soluble
in petroleum oils, and in carbon tetrachloride
Stability: stable at room temperature, but is decomposed by water
at higher temperatures and at pH 5.5 to form dichlorovos
Other names: trichlorphon (Great Britain), chlorofos (USSR), dipterex
(Turkey), metrifonate (WHO), Neguvon (veterinary use),
Dipterex, Tugon, Dylox, Bayer L 13/59, Bayer 15922
Reagents;
1. Trichlorfon standard of known % purity
2. Chloroform, pesticide or spectro grade preferred
3. Sodium sulfate, anhydrous, granular
-------
Trichlorfon EPA-1
(tentative)
Equipment;
1. Infrared spectrophotometer, double beam ratio recording,
with matched 0.2 mm NaCI or KBr cells
2. Mechanical shaker
3. Filtration apparatus or centrifuge
4. Usual laboratory glassware
Procedure:
Preparation of Standard;
Weigh 0,250 gram trichlorfon standard into a 25 ml volumetric
flask; dissolve and make to volume with chloroform. Add a small
amount of granular anhydrous sodium sulfate to insure dryness.
(final cone 10 mg trichlorfon/ml)
Preparation of Sample;
For dusts, granules, and wettable powders - weigh a portion of
sample equivalent to 0.5 gram trichlorfon into a 125 ml glass-
stoppered or screw-cap Erlenmeyer flask. Add 50 ml chloroform by
pipette and 1-2 grams anhydrous sodium sulfate. Close tightly,
shake on a mechanical shaker for 1 hour, allow to settle; filter
or centrifuge if necessary, taking precaution to avoid evaporation.
(final cone 10 mg trichlorfon/ml)
For liquids - weigh sample equivalent to 0.5 gram trichlorfon
into a 50 ml volumetric flask and make to volume with chloroform.
Add a small amount of anhydrous sodium sulfate to insure dryness.
IR Determination;
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument
being used, scan both the standard and sample from 1150 cm" to
900 cm"1 (8.7 p to 11.1 u).
-------
Trichlorfon EPA-1
(tentative)
Determine the absorbance of standard and sample using the peak
•I _T _ 1
at 1040 cm (9.6 u) and baseline from 1135 cm to 950 cm
(8.8 u to 10.53 p).
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent trichlorfon as follows:
. sample) (cone, std in mg/ml) (% purity std)
(abs. std) (cone, sample in mg/ml)
Method submitted by Mark W. Law, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
Any criticisms, suggestions, data, etc. on the use of this method will
be appreciated.
-------
April 1977 Trichlorfon EPA-2
(tentative)
Determination of Trichlorfon
by Gas Liquid Chromatography
(FID - Internal Standard)
Trichlorfon is the common name (approved by ISO) for dimethyl
(2,2,2-trichloro-l-hydroxyethyl) phosphonate, a registered insecti-
cide having the chemical structure:
CH-, O 0 OH Cl
CH3-0 (
—C—Cl
Molecular formula: C,HgCl30,P
Molecular weight: 257.5
Melting point: 83-84°C
Physical state, color, and odor: white crystalline solid
Solubility: 15.A g/100 ml water at 25°C; soluble in benzene, ether,
ethanol, and most chlorinated solvents; slightly soluble
in petroleum oils, and in carbon tetrachloride
Stability: stable at room temperature, but is decomposed by water
at higher temperatures and at pH 5.5 to form dichlorovos
Other names: trichlorphon (Great Britain), chlorofos (USSR), dipterex
(Turkey), metrifonate (WHO), Neguvon (veterinary use),
Dipterex, Tugon, Dylox, Bayer L 13/59, Bayer 15922
Reagents:
1. Trichlorfon standard of known % purity
2. Diethyl phthalate standard of known % purity
-------
Trichlorfon EPA-2
(tentative)
3. Acetone, pesticide or spectro grade
A. Internal Standard solution - weigh 0.5 gran diethyl phthalate
standard into a 50 ml volumetric flask; dissolve in and make
to volume with acetone, (cone 10 rag diethyl phthalate/ml)
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6* x 2 mm ID glass column packed with 5% SE-30 on
Chromosorb W DMCS 80/100 mesh (or equivalent column)
3. Precision liquid syringe: 5 or 10 pi
4. Mechanical shaker
5. Centrifuge
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 135 C
Injection temperature: 200°C
Detector temperature: 200°C
Carrier gas: Nitrogen or Helium
Carrier gas flow: 30 ml/rain (adjusted for specific GC)
Hydrogen pressure: 20 psl (adjusted for specific GC)
Air pressure: 30 psi (adjusted for specific GC)
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
-------
Trichlorfon EPA-2
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.2 gram trichlorfon standard into a 50 ml volumetric
flask. Add by pipette 5 ml of the internal standard solution,
make to volume with acetone, and shake thoroughly, (final cone
4 mg trichlorfon and 1 mg dlethyl phthalate/ml)
Preparation of Sample;
For dusts, granules, and wettable powders - weigh a portion
of sample equivalent to 0.2 gram trichlorfon into a small glass-
stoppered flask or screw-cap bottle. Add by pipette 5 ml of the
internal standard solution and 45 ml acetone. Close tightly
and shake 1 hr. on shaking machine, (final cone 4 mg trichlorfon
and 1 mg diethyl phthalate/ml)
For liquids - weigh a portion of sample equivalent to 0.2 gram
trichlorfon into a 50 ml volumetric flask. Add 5 ml internal
standard solution, and make to volume with acetone, (final cone
4 mg trichlorfon and 1 mg dlethyl phthalate/ml)
Determination;
Inject 5 ul of standard and, if necessary, adjust the instrument
parameters and the volume injected to give a complete separation
within a reasonable time and peak heights of from 1/2 to 3/4 full
scale. The elution order is trichlorfon, then diethyl phthalate.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of trichlorfon and diethyl
phthalate from both the standard-internal standard solution and
the sample-internal standard solution.
-------
4 Trichlorfon EPA-2
(tentative)
Determine the RF value for each Injection of the standard-
internal standard solution as follows and calculate the average:
I.S. « internal standard * diethy1 phthalate
m (wt. I.S.)(% purity I.S«)_(pk. ht. or area trichlorfon)
(wt. trichlorfon)(2 purity trichlorfon)(pk. ht. or area I.S.)
Determine the percent trichlorfon for each injection of the
sample-internal standard solution as follows and calculate the
average:
2 m (wt« I.S.)(% purity I.S.)(pk. ht. or area trichlorfon)
(wt. sample)(pk. ht. or area I.S.)(RF)
Method submitted by Mark W. Law, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
Any criticisms, suggestions, data, etc. on this method will be
appreciated.
-------
May 1977 Triethylene Glycol EPA-1
(tentative)
Determination of Triethylene Glycol
by Gas Liquid Chrotnatography
(TCD - Internal Standard)
Triethylene glycol is a registered disinfectant having the
chemical structure:
CH2—0— CH2~ CH2-OH
CH2 - 0 - CH2 ~ CH2~ OH
Molecular formula: C,Hn/0.
6 14 4
Molecular weight: 150.17
Boiling point: 285-287°C
Physical state, color, and odor: colorless, hygroscopic, practically
odorless liquid
Solubility: miscible with water, alcohol, benzene, toluene; sparingly
soluble in ether; practically insoluble in petroleum
ether
Stability: stable; hygroscopic
Other names: 2,2'-ethylenedioxybis(ethattol)
Reagents;
1. Triethylene glycol standard of known % purity
2. Ethyl hexanediol standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 3.5 grains ethyl hexanediol
standard into a 100 ml volumetric flask and make to volume
with acetone; mix well, (cone 35 mg ethyl hexanediol/ml)
-------
2 Triethylene Glycol EPA-1
(tentative)
Equipment;
1. Gas chroraatograph with thermal conductivity detector (TCD)
2. Column: A1 x I/A" glass packed with 3% XE-60 on 60/80
Chromosorb G AW DMCS (or equivalent column)
3. Precision liquid syringe: 10 ul
A. Usual laboratory glassware
Operating Conditions for TCP;
Column temperature: 1AO C
Injection temperature: 185 C
Detector temperature: 230 C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas flow: 30 ml/min
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.5 gram triethylene glycol standard into a 25 ml
volumetric flask, add 10 ml internal standard solution by pipette,
and make to volume with acetone; mix well. (final cone 20 ^ig
triethylene glycol and 1A jig ethyl hexanediol/jil)
-------
Triethylene Glycol EPA-1
(tentative)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.5 gram triethylene
glycol into a 25 ml volumetric flask, add 10 ml internal standard
solution, and make to volume with acetone; mix well, (final cone
20 ug propylene glycol and 14 fig ethyl hexanediol/jil)
Determination;
Inject 2-3 pi of standard-internal standard solution and, if
necessary, adjust the instrument parameters and the volume injected
to give a complete separation within a reasonable time and peak
heights from 1/2 to 3/4 full scale. The peak heights of triethylene
glycol and ethyl hexanediol should be nearly the same (definitely
within 25% of each other); if not, concentrations should be
adjusted accordingly. The elution order is ethyl hexanediol, then
triethylene glycol.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of triethylene glycol and
ethyl hexanediol from both the standard-internal standard solution
and the sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
IS « internal standard « ethyl hexanediol
TEG - triethylene glycol
RF (wt. IS)(% purity IS)(pk. ht. or area TEG)
(wt. TEG)(% purity TEG)(pk. ht. or area IS)
-------
Triethylene Glycol EPA-1
(tentative)
Determine the percent triethylene glycol for each injection of
sample-internal standard solution as follows and calculate the
average:
% . (wt» IS)(Z purity IS)(pk. ht. or area TEG)
(wt. sample)(pk. ht. or area IS)(RF)
Method submitted by Stelios Gerazounis, EPA Product Analysis Laboratory,
Region II, New York, N.Y.
Any comments, criticisms, suggestions, data, etc. concerning the use
of this method will be appreciated.
-------
May 1977 Vacor (trade name) EPA-1
(tentative)
Determination of Vacor (trade name)
by Ultraviolet Spectroscopy
Vacor is the trade name (a common name has not yet been approved)
for N-3-pyridylmethyl-N'-p-nitrophenylurea, a registered rodenticide
having the chemical structure:
H 0 H
CH2—- N— C—N (/ \V N02
Molecular formula: C .H._N,0,
Molecular weight: 272.27
Melting point: 223-225°C with decomposition
Physical state, color, and odor: odorless, light yellow powder
Solubility: extremely low in water; slightly soluble in pyridine and
dimethylformamide at 25°C; soluble in pyridine, dimethyl-
formamide, methyl cellosolve, and dimethyl sulfoxide at
80°C; insoluble in acetone, methanol, ethanol, isopropanol,
ethyl acetate, butyl acetate, butyl cellosolve, aceto-
nitrile, chlorobenzene, and toluene
Stability:
Other names: Vacor and RH-787 (Rohm & Haas)
-------
Vacor (trade name) EPA-1
(tentative)
Reagents;
1. Vacor standard of known % purity
2. Methanol, spectro or pesticide grade
(Note! 95% ethanol may also be used.)
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm silica cells
2. Mechanical shaker
3. Filtration apparatus
4. Usual laboratory glassware
Procedure;
Preparation of Standard;
Weigh 0.05 gram Vacor standard into a 50 ml volumetric flask,
make to volume with methanol, and shake to dissolve. If available,
an ultrasonic shaker will aid solution of the Vacor. (Note! Even
though under Solubility, Vacor is listed as insoluble in methanol,
it has been found that 0.1 gram will dissolve in 50 ml.) Pipette
10 ml into a 100 ml volumetric flask, make to volume, and mix
thoroughly. Pipette 10 ml of this solution into another 100 ml
volumetric flask, make to volume with methanol, and mix thoroughly.
(final cone 10 ug Vacor/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.1 gram Vacor into a
250 ml glass-stoppered flask or screw-cap bottle, add 100 ml
methanol by pipette, close tightly, and shake on a mechanical shaker
for one hour. (Note! For a 2% meal-type sample, one hour is more
-------
Vacor (trade name) EPA-1
(tentative)
than adequate.) Allow to settle; filter or centrifuge if necessary,
taking care to prevent loss due to evaporation. Pipette 10 ml into
a 100 ml volumetric flask, make to volume with methanol, and mix
thoroughly. Pipette 10 ml of this solution into another 100 ml
volumetric flask, make to volume with methanol, and mix thoroughly.
(final cone 10 jig Vacor/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 328 nm with methanol
in each cell. Scan both the standard and sample from 360 nm to
260 nm with methanol in the reference cell. Measure the absorbance
of both standard and sample at 328 nm.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent Vacor as follows:
„ z (abs. sample)(cone, std in pg/ml)(% purity std)
(abs. std) (cone, sample in jig/ml)
Method submitted by George B. Radan, EPA Product Analysis Laboratory,
Region II, New York, N.Y.
Any comments, criticism, suggestions, data, etc. concerning the use of
this method will be appreciated.
-------
May 1977 Vacor (trade name) EPA-2
(tentative)
Determination of Vacor by
High Pressure Liquid Chromatography
Vacor is the trade name (a common name has not yet been approved)
for N-S-pyridylmethyl-N'-p-nitrophenylurea, a registered rodenticide
having the chemical structure:
Molecular formula: ci3Hi2N4°3
Molecular weight: 272.27
Melting point: 223-225°C with decomposition
Physical state, color, and odor: odorless, light yellow powder
Solubility: extremely low in water; slightly soluble in pyridine and
dimethylformamide at 25 C; soluble in pyridine, dimethyl-
formaraide, methyl cellosolve, and dimethyl sulfoxide at
80 C; insoluble in acetone, methanol, ethanol, Isopropanol,
ethyl acetate, butyl acetate, butyl cellosolve, aceto-
nitrlle, chlorobenzene, and toluene
Stability:
Other names: Vacor and RH-787 (Rohm & Haas)
-------
Vacor (trade name) EPA-2
(tentative)
Reagents;
1. Vacor standard of known % purity
2. Dimethylformaraide, spectro or pesticide grade
Equipment;
1. High pressure liquid chromatograph with variable wavelength
UV detector (for 327 run)
2. Suitable column such as: DuPont Permaphase ETH, 0.5 meter
x 2.1 mm I.D.
(Permaphase ETH is an ether stationary phase chemically
bonded to the surface of "Zipax." Permaphase ETH is a polar
bonded packing by means of a Si-O-Si bond. The support
contains approx. 12 stationary phase by weight.)
3. 5 pi high pressure liquid syringe or sample injection loop
4. Mechanical shaker
5. Centrifuge
6. 5 micron mllllpore filter
7. Usual laboratory glassware
Operating Conditions;
Mobile phase: 15% msthanol + 85% water
Column temperature: ambient
Flow rate: 0.75 to 1.0 ml/min
Chart speed: 12'/hi:
Amount injected: 5 pi
Conditions may have to be adjusted for the specific instrument
being used, column variations, sample composition, etc. to obtain
optimum response and reproducibility.
-------
Vacor (trade name) EPA-2
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.05 gram Vacor into a 50 ml volumetric flask; dissolve
in and make to volume with dinethylformamide; mix thoroughly.
(final cone 1 mg Vacor/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.1 gram Vacor into
a 250 ml glass-stoppered flask or screw-cap bottle, add 100 ml
dimethylforraamide by pipette, close tightly, and shake for one
hour on a mechanical shaker. Allow to settle; if not crystal
clear, centrifuge a portion for 5 minutes. If still not crystal
clear, filter through a 5 micron mlllipore filter. Take pre-
caution to prevent evaporation, (final cone 1 mg Vacor/ml)
Determination;
Alternately inject three 5 ;il portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the percent
Vacor as follows:
. m (pk. ht. or area sample)(wt. std injected)(Z purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
Any comments, criticism, suggestions, data, etc. concerning the use of
this method will be appreciated.
-------
I
-------
Analytical Methods - Second Supplement
May 1, 1979
1. Bendiocarb EPA-1 (tentative)
2. Bendiocarb EPA-2 (tentative)
3, Butylate EPA-6 (tentative)
4. Carboxin EPA-2 (tentative)
5. Chlorothalonil EPA-2 (tentative)
6. Chlorpyrifos EPA-1 (tentative)
7. Chlorpyrifos EPA-2 (tentative)
8. Chlorpyrifos EPA-3 (tentative)
9. Chlorpyrifos EPA-4 (tentative)
10. Deet EPA-4 (tentative)
11. Diphacinone EPA-2 (tentative)
12. Diphenylamine EPA-1 (tentative)
13. Endosulfan EPA-5 (tentative)
14. EPTC EPA-6 (tentative)
15. Ethofumesate EPA-4 (tentative)
16. Flammability Test EPA-1 (Elame Projection)
17. Elammability Test EPA-2 (Drum Test)
18. Eluometuron EPA-2 (tentative)
19. Methomyl EPA-1 (tentative)
20. Methoxychlor EPA-3 (tentative)
21. Mexacarbate EPA-1 (tentative)
22. Mixed Pesticides EPA-1 (Warfarin & Sulfaquinoxaline)
23. Oryzalin EPA-1 (tentative)
24. Parathion EPA-3 (tentative)
25. Parathion EPA-4 (tentative)
26. Pirimicarb EPA-1 (tentative)
27. Pyrazon EPA-1 (tentative)
28. Strychnine EPA-3 (tentative)
29. Vernolate EPA-4 (tentative)
30. TLC Identification EPA-2
-------
-------
Analytical Methods - Second Supplement
May 1, 1979
Pesticide Name Cross Reference Index to the Methods
4~amino-N-2-quinoxalinylbenzene-
sulfonamide
Bendiocarb EPA-1 (tentative)
Bendiocarb EPA-2 (tentative)
Big Dipper
Butylate EPA-6 (tentative)
Carboxin EPA-2 (tentative)
Chlorothalonil EPA-2 (tentative)
Chlorpyrifos EPA-1 (tentative)
Chlorpyrifos EPA-2 (tentative)
Chlorpyrifos EPA-3 (tentative)
Chlorpyrifos EPA-4 (tentative)
Compound 3-120
Deet EPA-4 (tentative)
2-dimethylamino-5,6-dimethyl-
pyrimidin-4-yl-dimethylcarbamate
4-dimethylamino-3,5-xylyl
N-methylcarbamate
2,2-dimethyl-l,3-benzodioxol-
4-yl-N-methylcarbamate
5,6-dimethyl-2-dimethylamino-
4-pyrimidinyl dimethylcarbamate
Diphacinone EPA-2 (tentative)
Diphenylamine EPA-1 (tentative)
Dirimal
Mixed Pesticides EPA-1
(see sulfaquinoxaline)
IR
UV
Diphenylamine EPA-1 (tentative)
HPLC - reversed phase
UV
GLC-FID-IS
IR
UV
GLC-TCD-IS
HPLC
Mixed Pesticides EPA-1
(see sulfaquinoxaline)
HPLC - normal phase
Pirimicarb EPA-1 (tentative)
Mexacarbate EPA-1 (tentative)
Bendiocarb EPA-1 (tentative)
Pirimicarb EPA-1 (tentative)
HPLC-PIC
GLC-TCD
Oryzalin EPA-1 (tentative)
-------
Douco 139
Dou/co 179
Dursban (mosquito control)
EL-119
Endosulfan EPA-5 (tentative)
EPIC EPA-6 (tentative)
Ethofumesate EPA-1 (tentative)
Ficam
Flammability Test EPA-1
Flammability Test EPA-2
Fluometuron EPA-2 (tentative)
Garvox
H 119
2,3-isopropylidenedioxypheny1
methylcarbamate
Lannate
Lorsban (agricultural use)
Me thorny 1 EPA-1 (tentative)
Methoxychlor EPA-3 (tentative)
Methylcarbamic acid 4-
(dimethylamino)-3,5-xyly.l ester
Mexacarbate, EPA-1 (tentative)
Mixed Pesticides EPA-1
(Warfarin & sulfaquinoxaline)
Multimet
NC 6897
Nortron
Mexacarbate EPA-1 (tentative)
Chlorpyrifos EPA-1, 2, 3, 4
Chlorpyrifos EPA-1, 2, 3, 4
Oryzalin EPA-1 (tentative)
GLC-EID-IS
HPLC - reversed phase
GLC-FID-IS
Bendiocarb EPA-1 (tentative)
Elame Projection
Closed Drum
UV
Bendiocarb EPA-1 (tentative)
Pyrazon EPA-1 (tentative)
Bendiocarb EPA-1 (tentative)
Methomyl EPA-1 (tentative)
Chlorpyrifos EPA-1, 2, 3, 4
HPLC
HPLC - normal phase
Mexacarbate EPA-1 (tentative)
GLC-TCD-IS
HPLC-PIC
Bendiocarb EPA-1 (tentative)
Bendiocarb EPA-1 (tentative)
Ethofumesate EPA-1 (tentative)
-------
Nudrin
Oryzalin EPA-1 (tentative)
Parathion EPA-3 (tentative)
Parathion EPA-4 (tentative)
PCA
N-phenylbenzeneamine'
Pirimicarb EPA-I (tentative)
Pirimor
PP062
Pyramin
Pyrazon EPA-1 (tentative)
Ryzelan
Scaldip
Strychnine EPA-3 (tentative)
Sulfabenzpyrazine
Sulfacox
Sulfaline
2-sulfanilamidoquinoxaline
Sulfa-Q
Sulfaquinoxaline (with Warfarin)
Sulquin
Surflan
Methomyl EPA-1 (tentative)
Visible (colorimetric) spectroscopy
GLC-FID-I5
HPLC - reversed phase
Pyrazon EPA-1 (tentative)
Diphenylamine EPA-1 (tentative)
UV
Pirimicarb EPA-1 (tentative)
Pirimicarb EPA-1 (tentative)
Pyrazon EPA-1 (tentative)
UV
Oryzalin EPA-1 (tentative)
Diphenylamine EPA-2 (tentative)
HPLC - reversed phase
Mixed Pesticides EPA-1
(see sulfaquinoxaline)
Mixed Pesticides EPA-1
(see sulfaquinoxaline)
Mixed Pesticides EPA-1
(see sulfaquinoxaline)
Mixed Pesticides EPA-1
(see sulfaquinoxaline)
Mixed Pesticides EPA-1
(see sulfaquinoxaline)
Mixed Pesticides EPA-1
Mixed Pesticides EPA-1
(see sulfaquinoxaline)
Oryzalin EPA-1 (tentative)
-------
TIC Identification EPA-2
Trichloropyrphos
Vernolate EPA-4 (tentative)
Warfarin (with sulfaquinoxaline)
Zectran
TLC
Chlorpyrifos EPA-1, 2, 3, 4
HPLC - reversed phase
Mixed Pesticides EPA-1
Mexacarbate EPA-1 (tentative)
-------
March 1978
Bendiocarb EPA-1
(tentative)
The Determination of Bendiocarb in
Wettable Powder Formulations by Infrared Spectroscopy
Bendiocarb is the common name for 2,2-dimethyl-l,3-benzodioxol-
4-yl N-methylcarbamate, a registered insecticide having the chemical
structure:
,
/
CH3
0 C-
II
0
NH—CH3
Molecular formula:
Molecular weight: 223.23
Melting point: 129-130°C
Physical state, color, and odor:
white crystalline solid
Solubility:
Stability:
Other names:
at 25°C is: 0.004% in water, 0.03% in kerosene, 1.0% in
o-xylene, 4% in ethanol and benzene, and 20% in acetone,
dichloromethane, dioxan and chloroform
the hydrolysis (to the phenol) half-life in solution in
0.01M aqueous sodium phosphate buffer at pH 7 and 25 C
is 20 days.
Ficam (Fisons Ltd., Great Britain); NC 6897; Garvox;
Multimet; 2,3-isopropylidenedioxyphenyl methylcarbamate
Reagents;
1. Bendiocarb standard of known % purity
2. Chloroform, spectro or pesticide grade
3. Sodium sulfate, anhydrous, granular
Equipment;
1. Infrared spectrophotometer, double beam ratio recording
with matched 0.5 mm NaCl or KBr cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Hypodermic syringe (1-2 ml, for filling IR cells)
-------
Bendiocarb EPA-1
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.1 gram bendiocarb standard into a small glass-
stoppered flask or screw-cap bottle, add 25 ml chloroform by
pipette, and shake to dissolve. Add a small amount of anhydrous
sodium sulfate to insure dryness. (cone 4 mg/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.2 gram bendiocarb
into a 125 ml glass-stoppered flask or screw-cap bottle. Add
50 ml chloroform by pipette and 1-2 grams anhydrous sodium
sulfate. Close tightly and shake on a mechanical shaker for 30
minutes. Allow to settle, centrifuge or filter if necessary,
taking precaution to prevent evaporation. (final cone 4 mg/ml)
IR Determination;
With chloroform in the reference cell, and using the optimum
quantitative analytical settings for the particular infrared
spectrophotometer being used, scan the standard and sample from
2000 cm'1 to 1538 cm'1 (5.0 u to 6.5 u).
Determine the absorbance of standard and sample using the
peak at 1761 cm'1 (5.68 u) and a baseline from 1961 cm" to
1703 cm (5.10 u to 5.87 u).
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent bendiocarb as follows:
7 - (a>s. sample) (cone, std in mg/ml) (% purity, of std)
(abs. standard)(cone, sample in mg/ml)
Method submitted by Stelios Gerazounis, EPA Chemistry Laboratory,
Region II, New York.
Any comments, criticism, suggestion, data, etc. concerning the use
of this method will be appreciated.
-------
June 1978
Bendiocarb EPA-2
(tentative)
The Determination of Bendiocarb in
Wettable Powder Formulations by Ultraviolet Spectrometry
Bendiocarb is the common name for 2,2-dimethyl-l,3-benzodioxol-
4-yl N-methylcarbamate, a registered insecticide having the chemical
structure:
Molecular formula: C1;LH13N04
Molecular weight: 223.23
Melting point:
129-130°C
Physical state, color, and odor: white crystalline solid
Solubility: at 25°C is: 0.004% in water, 0.03% in kerosene, 1.0% in
o-xylene, 4% in ethanol and benzene, and 20% in acetone,
dichloromethane, dioxan and chloroform
Stability: the hydrolysis (to the phenol) half-life in solution in
0.01M aqueous sodium phosphate buffer at pH 7 and 25 C
is 20 days.
Other names: Ficam (Fisons Ltd., Great Britain); NC 6897; Garvox;
Multimet; 2,3-isopropylidenedioxyphenyl methylcarbamate
Reagents;
1. Bendiocarb standard of known % purity
2. Methanol, spectro or pesticide grade
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm silica cells
2. Usual laboratory glassware
-------
Bendlocarb EPA-2
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.1 gram bendiocarb standard into a 100 ml volumetric
flask. Dissolve in, make to volume with methanol, and mix
thoroughly. Pipette 5 ml into a second 100 ml volumetric flask,
make to volume with methanol, and mix thoroughly. (cone 50 ug/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.1 gram bendiocarb
into a 100 ml volumetric flask. Make to volume with methanol and
mix thoroughly. Pipette 5 ml into a second 100 ml volumetric
flask, make to volume with methanol, and mix thoroughly. (final
cone 50 ug/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 278 nm with
methanol in each cell. Scan both the standard and sample from
310 nm to 240 nm with methanol in the reference cell. Measure
the absorbance of both standard and sample at 278 nm.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent bendiocarb as follows:
(abs. sample)(cone, standard in ug/ml)(% purity standard)
(abs. standard)(cone, sample in ug/ml)
Note! This method is linear from 0 to 250 ug/ml final concentration.
Method submitted by George B. Radan, EPA Chemistry Laboratory,
Region II, New York.
Any criticism, data, or suggestions concerning this method will be
appreciated.
-------
March 1978 Butylate EPA-6
(tentative)
Determination of Butylate by
High Pressure Liquid Chromatography
(Reverse Phase)
Butylate is the common name for S-ethyl diisobutylthiocarbamate,
a registered herbicide having the chemical structure:
0
II
CH3—CH2 S—C N
CH2 CH CH3
CH2 CH CH3
Molecular formula: C -H__ONS CH3
'11 23
Molecular weight: 217.4
Boiling point: 71 at 10 mm
Physical state and color: Amber liquid
Solubility: 45 ppm in water at room temperature; miscible with
kerosene, acetone, methyl isobutyl ketone, ethanol, xylene
Stability: stable under ordinary conditions; non-corrosive
Other names: Sutan (Stauffer), R1910
Reagents;
1. Butylate standard of known % purity
2. Dioxane, pesticide or spectro grade
Equipment;
1. High pressure liquid chromatograph with UV detector at 254 nm.
If a variable wavelength UV detector is available, other wave-
lengths may be used to increase sensitivity or to eliminate
interference; 230 nm is very good for butylate.
2. Suitable column such as:
a. Dupont ODS Permaphase, 1 meter x 2.1 mm ID
b. Perkin-Elmer ODS Sil-X-II RP, (two) 0.5 meter x 2.6 mm ID
3. High pressure liquid syringe or sample injection loop
4. Usual laboratory glassware
-------
2 Butylate EPA-6
(tentative)
Operating Conditions;
Mobile phase: 25% acetonitrile 4- 75% water
Column temperature: 30°C
Chart speed: 12"/hr
Flow rate: 0.5 to 1.5 ml/min (Perkin-Elmer 1 meter column)
Pressure: 1800-2000 psi (Dupont 1 meter column)
Attenuation: Adjusted
Conditions may have to be varied by the analyst for the specific
instrument being used, column variations, sample composition, etc. to
obtain optimum response and reproducibility.
Procedure:
Preparation of Standard;
Weigh 0.1 gram butylate standard into a 50 ml volumetric flask,
make to volume with dioxane, and mix thoroughly. (cone 2 rag/ml)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.1 gram butylate
into a 50 ml volumetric flask, make to volume with dioxane, and
mix thoroughly. (cone 2 mg butylate/ml)
Determination;
Alternately, inject three 5 ul portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the
percent butylate as follows:
7 = (pk» ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Note; Generally, butylate can be easily separated on a reverse phase
HPLC system if the right solvent polarity is used. Acetonitrile
and methanol are good primary solvents. They have similar
dielectric constants but have different hydrogen bondings. Since
butylate's molecular weight is high, acetonitrile would be the
solvent of choice because of its low polarity. This would give
a short retention time with good resolution. If more separation
is desired, methanol should be used, but some loss of resolution
would be expected.
Method submitted by Elmer H. Hayes, EPA Beltsville Chemistry Laboratory,
Beltsville, Maryland.
Any criticism, data, or suggestions concerning the use of this method
will be appreciated.
-------
March 1978
Carboxin EPA-2
(tentative)
Determination of Carboxin in Dusts
and Powders by Ultraviolet Spectroscopy
Carboxin is the common name for 5,6-dihydro-2-methyl-l,4-
oxathiin-3-carboxanilide, a registered fungicide having the chemical
structure:
Molecular formula:
Molecular weight:
Melting point:
235
91.5 to 92.5 C; a dimorphic form has a m.p. of
98 to 100°C
Physical state, color, and odor: odorless, white, crystalline solid
(The technical product is at least 97% pure.)
Solubility: 170 ppm in water at 25 C; soluble in acetone, benzene,
dimethyl sulfoxide, ethanol, methanol
Stability: compatible with all except highly alkaline or acidic pesticides
Other names: Vitavax, D735 (Uniroyal); DCMO
Reagents;
1. Carboxin standard of known % purity
2. Methanol, pesticide or spectro grade
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Filtration apparatus
4. Usual laboratory glassware
Procedure;
Preparation of Standard;
Weigh 0.1 gram carboxin standard into a 100 ml volumetric
flask. Dissolve in, make to volume with methanol, and mix thor-
oughly. Pipette a 10 ml aliquot into a second 100 ml volumetric
flask and make to volume, with methanol. Mix thoroughly and pipette
a 10 ml aliquot into a third 100 ml volumetric flask. Make to
volume with methanol and again mix thoroughly. (cone 10 ug/ml)
-------
2 Carboxin EPA-2
(tentative)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.1 gram of carboxin
into a 250 ml Erlenmeyer glass-stoppered flask. Add 100 ml
methanol by pipette and shake on a mechanical shaker for one
hour. Allow to settle; filter if necessary. Pipette 10 ml of
the clear filtrate into a 100 ml volumetric flask, make to
volume with methanol, and mix thoroughly. Pipette 10 ml into
another 100 ml volumetric flask, make to volume with methanol,
and again mix thoroughly. (final cone 10 ug carboxin/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 248 nm with
methanol in each cell. Scan both the standard and sample from
300 nm to 200 nm with methanol in the reference cell. Measure
the absorbance of both standard and sample at 248 nm.
Calculation;
From the above absorbances and using the standard and
sample concentrations, calculate the percent carboxin as
follows;
7 = (abs. sample)(cone, stdin ug/ml)(% purity std)
(abs. std)(cone, sample in ug/ml)
Method submitted by Edward Zager, EPA Product Analysis Laboratory,
Region II, New York, NY.
Any criticism, data, or suggestions concerning the use of this method
will be appreciated.
-------
December 1978
Chlorothalonil EPA-2
(tentative)
Determination of Chlorothalonil
by Gas Liquid Chromatography
(FID - Internal Standard)
Chlorothalonil is the common name for tetrachloroisophthalonitrile,
a registered fungicide having the chemical structure:
0=N
Cl
Molecular formula:
Molecular weight:
Melting point:
266
250 to 251°C
Physical state, color, and odor: white crystalline solid, odorless in
pure form; the technical product (about 98% pure)
has a slightly pungent odor.
Solubility:
Stability:
Other names:
insoluble in water (0.6 ppm); slightly soluble in acetone
(2% w/w), cyclohexanone (3?o w/w), methyl ethyl ketone
(2% w/w), xylene (8?o w/u/) , and kerosene less than 1%
stable to ultraviolet radiation and to moderately alkaline
and acid aqueous media; thermally stable under normal
storage conditions; non-corrosive
Daconil 2787 (Diamond Shamrock Chem. Co.); Bravo; Termil;
2,4,5,6-tetrachloro-l,3-dicyanobenzene; 2,4,5,6-tetrachloro-
3-cyanobenzonitrile
Reagents;
1.
2.
3.
Chlorothalonil standard of known % purity
Aldrin standard of known HHDN content
Xylene, pesticide or spectre grade preferred, ACS ok
Note: large injections of xylene may dirty the detector.
Internal standard solution - weigh 0.15 gram aldrin into a
100 ml volumetric flask, dissolve in and make to volume u/ith
xylene. Mix well, (cone 1.5 mg/ml or 1.5 ug/ul)
-------
2 Chlorothalonil EPA-2
(tentative)
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm ID glass column packed with 1% XE-60 on
80/100 mesh Chromosorb G (or equivalent column)
3. Precision liquid syringe: 1 or 5 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 170 C
Injection temperature: 300 C
Detector temperature: 300°C
Carrier gas: Nitrogen 2
Carrier gas pressure: Adjust for optimum performance (0.9 Kg/cnu)
Hydrogen pressure: Adjust for optimum performance (0.7 Kg/crru)
Air pressure: Adjust for optimum performance (1.3 Kg/cm )
Chart speed: 0.25"/tnin or 15"/hr
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
Procedure;
Preparation of Standard;
Weigh 0.1 gram chlorothalonil standard into a small glass-
stoppered flask or screw-cap tube. Add by pipette 20 ml of the
internal standard solution and shake to dissolve, (final cone
5 mg chlorothalonil and 1.5 mg aldrin/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.1 gram chlorothalonil
into a small glass-stoppered flask or screw-cap tube. Add by pipette
20 ml of the internal standard solution. Close tightly and shake on a
mechanical shaker to dissolve. Filter if necessary, (final cone
5 mg chlorothalonil and 1.5 mg aldrin/ml)
Determination;
Inject 0.5-1 ul of standard and, if necessary, adjust the instru-
ment parameters and the volume injected to give a complete separation
within a reasonable time and peak heights of from 1/2 to 3/4 full
scale. The elution order is aldrin, then chlorothalonil.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of chlorothalonil and aldrin
from both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
-------
Chlorothalonil EPA-2
(tentative)
chlor. = chlorothalonil
(\i/t. aldrin)(?o purity aldrin)(pk. ht. or area chlor.)
ptr —
(wt. chlor .~5(?c purity chlor.)(pk. ht. or area aldrin)
Determine the percent chlorothalonil for each injection of
the sample-internal standard solution as follows and calculate
the average:
wt. aldrin) (?o purity aldrin)(pk. ht. or area chlor.)
u/t. sample)(pk. ht. or area aldrin) (RF)
Method submitted by Dr. Gabriele Tartari, Agrochemical Dept., Control
Laboratory, CIBA-GEIGY S.p.A., C.P. 88, 1-21047 Saronno (VA), ITALY.
Any criticism, data or suggestions concerning the use of this method
will be appreciated.
-------
January 1979
Chlorpyrifos EPA-1
(tentative)
Determination of Chlorpyrifos
by Infrared Spectroscopy
Chlorpyrifos is the accepted (ANSI, ISO, BSI) common name for
0,0-diethyl Q-(3,5,6-trichloro-2-pyridyl)-phosphorothioate, a
registered insecticide having the chemical structure:
• i
-o-p
\
0-CH2—CH3
0-CH2-CH3
Molecular formula: CgH,,C13NCLPS
Molecular weight: 350.5
Physical state, color, and odor:
white crystals with a mild mercaptan
odor
Melting point:
Solubility
41 to 43UC
2 ppm in water at 25 C; 79% in isooctane w/w, 43?o in methanol
w/w; readily soluble in most other organic solvents
Stability: stable under normal storage conditions; compatible with
non-alkaline pesticides but is corrosive to copper and brass;
half-life in agueous methanolic solution 1930 days at pH 6.0,
7.2 days at pH 9.96
Other names: Dursban (for mosguito control); Lorsban (for agricultural use);
Dowco 179 (Dow Chemical); trichlorpyrphos
Reagents;
1. Chlorpyrifos standard of known % purity
2. Carbon disulfide, pesticide or spectro grade preferred
3. Sodium sulfate, anhydrous, granular
Eguipment;
1. Infrared spectrophotometer, double beam ratio recording, with
matched 0.5 mm NaCl or KBr cells
2. Mechanical shaker
3. Filtration apparatus or centrifuge
4. Usual laboratory glassware
-------
Chlorpyrifos EPA-1
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.125 gram chlorpyrifos standard into a 25 ml volumetric
flask, dissolve in and make to volume with carbon disulfide; mix
thoroughly. Add a small amount of granular anhydrous sodium sulfate
to insure dryness. (final cone 5 mg chlorpyrifos/ml)
Preparation of Sample;
For dusts, 'granules, and wettable powders - weigh a portion of
sample equivalent to 0.25 gram chlorpyrifos into a 125 ml glass-
stoppered or screw-cap Erlenmeyer flask. Add 50 ml carbon disulfide
by pipette and 1-2 grams anhydrous sodium sulfate. Close tightly,
shake on a mechanical shaker for 1 hour, allow to settle, filter
or centrifuge if necessary taking precaution to avoid evaporation.
(final cone 5 mg chlorpyrifos/ml)
For liquids - weigh sample equivalent to 0.25 gram chlorpyrifos
into a 50 ml volumetric flask and make to volume with carbon disulfide.
Add a small amount cf anhydrous sodium sulfate to insure dryness.
(final cone 5 mg chlorpyrifos/ml)
IR Determination;
With carbon disulfide in the reference cell, and using the optimum
quantitative analytical settings for the particularlR instrument being
used, scan both the standard and sample from 990 cm" to 900 cm" (10.1 u
to 11.1 u).
Determine the absorbance of standard and sample using the peak
at 960 cm" (10.4 u) and a horizontal baseline from 930 cm (10.75 u).
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent chlorpyrifos as follows:
D' _ (abs. sample) (cone, std in mq/ml)(?o purity std)
Tabs.std)(cone.sample in mg/ml)
Method submitted by Mississippi State Chemistry Laboratory, Box CR,
Mississippi State, Mississippi 39762.
Any criticisms, suggestions, data, etc. on the use of this method will
be appreciated.
-------
January 1979
Chlorpyrifos EPA-2
(tentative)
Determination of Chlorpyrifos
by Ultraviolet Spectroscopy
Chlorpyrifos is the accepted (ANSI, ISO, BSI) common name for
0,0-diethyl 0-(3,5,6-trichloro-2-pyridyl)-phosphorothioate, a
registered insecticide having the chemical structure:
| ,0-CH2-CH3
•0-P
—CH3
Molecular formula: CgH1 ..Cl^NO^P!
Molecular weight: 350.5
Physical state, color, and odor:
Melting point: 41 to 43°C
white crystals with a mild mercaptan
odor
Solubility:
Stability:
2 ppm in water at 25 C; 19% in isooctane w/w, 43?o in methanol
w/w; readily soluble in most other organic solvents
stable under normal storage conditions; compatible with
non-alkaline pesticides but is corrosive to copper and brass;
half-life in aqueous methanolic solution 1930 days at pH 6.0,
7.2 days at pH 9.96
Other names: Dursban (for mosquito control); Lorsban (for agricultural use);
Dowco 179 (Dow Chemical); trichlorpyrphos
Reagents:
1. Chlorpyrifos standard of known % purity
2. Methanol, pesticide or spectro grade
Equipment:
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm silica cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
-------
Chlorpyrifos EPA-2
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.1 gram chlorpyrifos standard into a 100 ml volumetric
flask, dissolve in and make to volume with methanol; mix thoroughly.
Pipette 10 ml into a second 100 ml volumetric flask, make to volume
with methanol, and mix thoroughly. Pipette 20 ml of this solution
into a third 100 ml volumetric flask and make to volume with methanol,
Mix thoroughly, (final cone 20 ug chlorpyrifos/ml)
Preparation of Sample;
For wettable powders - weigh a portion of sample equivalent to
0.1 gram chlorpyrifos into a 250 ml glass-stoppered flask or screw-
cap bottle. Add 100 ml of methanol, close tightly, and shake for
30 minutes. Allow to settle, centrifuge or filter if necessary.
Proceed as in the third paragraph beginning "Pipette 10 ml - - -."
For liquid formulations - weigh a portion of sample equivalent
to 0.1 gram chlorpyrifos into a 100 ml volumetric flask, make to
volume with methanol, and mix thoroughly. Proceed as below.
Pipette 10 ml of either of the above sample solutions into a
100 ml volumetric flask, make to volume with methanol, and mix
thoroughly. Pipette 20 ml of this solution into a 100 ml volumetric
flask, make to volume with the methanol, and mix thoroughly, (final
cone 20 ug chlorpyrifos/ml)
UV Determination;
Balance pen for 0 and 100?o transmission at 285 nm with methanol
in each cell. Scan standard and sample from 330 nm to 230 nm with
methanol in the reference cell. Measure absorbance of standard and
sample at 285 nm.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent chlorpyrifos as follows:
0, _ (abs. sample) (cone, std in uq/ml)(/n purity std)
(abs. std)(cone, sample in ug/ml)
Method submitted by David Person, EPA Product Analysis Laboratory,
Region II, New York, NY.
Any criticisms, suggestions, or data concerning the use of this method
will be appreciated.
-------
April 1979
Chlorpyrifos EPA-3
(tentative)
Determination of Chlorpyrifos
by Gas Liquid Chromatography
(TCD - Internal Standard)
Chlorpyrifos is the accepted (ANSI, ISO, BSI) common name for
0,0-diethyl Q-(3,5,6-trichloro-2-pyridyl)-phosphorothioate, a
registered insecticide having the chemical structure:
-Cl
]|
0-P
C9H11C13N03PS
Molecular formula:
Molecular weight: 350.5
Physical state, color, and odor: white crystals with a mild mercaptan odor
Melting point:
41 to 43 C
Solubility: 2 ppm in water at 25 C; 79% in isooctane w/w, 43?o in methanol
w/w; readily soluble in most other organic solvents
Stability: stable under normal storage conditions; compatible with
non-alkaline pesticides but is corrosive to copper and brass;
half-life in aqueous methanolic solution 1930 days at pH 6.0,
7.2 days at pH 9.96
Other names: Dursban (for mosquito control); Lorsban (for agricultural use);
DOM/CO 179 (Dou/ Chemical); trichlorpyrphos
Reagents;
1. Chlorpyrifos standard of known % purity
2. Benzyl benzoate standard of known % purity
3. Chloroform, pesticide or spectro grade
4. Internal Standard solution - weigh 1.3 gram benzyl benzoate
standard into a 100 ml volumetric flask, dissolve in and make to
volume with chloroform; mix well, (cone 13 mg benzyl benzoate/ml)
Equipment;
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 4' x 1/4" ID glass, packed with b% SE-30 on 80/100
Diatoport S or equivalent column (such as 4' x 1/4" ID glass,
packed with 4?o SP-2100 on 80/100 Chromosorb 750)
3. Precision liquid syringe: 10 ul
4. Usual laboratory glassware
-------
Chlorpyrifos EPA-3
(tentative)
Operating Conditions for TCP;
Column temperature:
180°C
Injection temperature; 215 C
Detector temperature: 230 C
Filament current:
Carrier gas:
Carrier gas pressure:
200 ma
Helium
40 psi (20 ml/min)
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
Procedure;
Preparation of Standard;
For use with emulsifiable concentrates and liquid formulations -
weigh 0.15 gram chlorpyrifos standard into a 25 ml volumetric flask;
add 5 ml internal standard solution by pipette, make to volume with
chloroform, and mix well, (final cone 6 mg chlorpyrifos and 2.6 mg
benzyl benzoate/ml)
For use with dusts, granules, and wettable powders - weigh 0.15
gram chlorpyrifos standard into a small glass-stoppered flask or
screw-cap bottle, add by pipette 5 ml of internal standard solution
and 25 ml chloroform, close tightly and shake well to dissolve the
chlorpyrifos. (final cone 6 mg chlorpyrifos and 2.6 mg benzyl
benzoate/ml)
Preparation of Sample;
For gmulsifiable concentrates and liquid formulations - weigh a
portion of sample equivalent to 0.15 gram chlorpyrifos into a 25 ml
volumetric flask; add 5 ml of internal standard solution by pipette,
make to volume with chloroform, and mix well, (final cone 6 mg
chlorpyrifos and 2.6 mg benzyl benzoate/ml)
For dusts, granules, and wettable powders - weigh a portion of
sample equivalent to 0.15 gram chlorpyrifos into a small glass-
stoppered flask or screw-cap bottle; add by pipette 5 ml internal
standard and 25 ml of chloroform, close tightly and shake on a
mechanical shaker for 10-15 minutes or shake by hand intermittently
for 25-30 minutes, (final cone 6 mg chlorpyrifos and 2.6 mg benzyl
benzoate/ml)
Determination;
Inject 2 ul of standard and, if necessary, adjust the instrument
parameters and the volume injected to give a complete separation
within a reasonable time and peak heights of from 1/2 to 3/4 full
scale. The elution order is benzyl benzoate, then chlorpyrifos.
Proceed with the determination, making at least three injections
each of standard and sample solutions in random order.
-------
3 Chlorpyrifos EPA-3
(tentative)
Calculation;
Measure the peak heights or areas of chlorpyrifos and benzyl
benzoate from both the standard-internal standard solution and
the sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(I.S. = benzyl benzoate)
(wt. I.S.)(?o purity I.S.)(pk. ht. or area chlorpyrifos) _
(wt. chlorpyrifos) '(% purity chlorpyrifos) (pk. ht. or area I.S.)
Determine the percent chlorpyrifos for each injection of the
sample-internal standard solution as follows and calculate the
average:
. I»S.)(?o purity I.S.)(pk. ht. or area chlorpyrifos)
(u/t. sample)(pk. ht. or area I.S.)(RF)
Method submitted by Stelios Gerazounis, EPA Product Analysis Lab, Region II,
New York, NY. (also from experimental #17 method May 1970)
Any criticisms, suggestions, or data concerning this method or its use
will be appreciated.
-------
April 1979
Chlorpyrifos EPA-4
(tentative)
Determination of Chlorpyrifos by
High Pressure Liquid Chromatography
Chlorpyrifos is the accepted (ANSI, ISO, BSI) common name for
0,0-diethyl 0-(3,5,6-trichloro-2-pyridyl)-phosphorothioate, a
registered insecticide having the chemical structure:
•C!
S
II
•o—p.
Molecular formula: C-H.^Cl.jNO.j
Molecular weight: 350.5
Physical state, color, and odor
Melting point:
Solubility
white crystals with a mild mercaptan odor
to 43°C
Stability;
Other names:
2 ppm in water at 25 C; 79% in isooctane w/w, 43?o in methanol
w/w; readily soluble in most other organic solvents
stable under normal storage conditions; compatible with
non-alkaline pesticides but is corrosive to copper and brass;
half-life in aqueous methanolic solution 1930 days at pH 6.0,
7.2 days at pH 9.96
Dursban (for mosquito control); Lorsban (for agricultural use!
Dowco 179 (Dow Chemical); trichlorpyrphos
Reagents;
1. Chlorpyrifos standard of known % purity
2. Methanol, spectro or pesticide grade
Equipment;
1. High pressure liquid chromatograph with variable wavelength UV
detector (for 289 nm)
2. Suitable column such as: Partisil 10 ODS 25 cm x 4.6 mm ID
3. 10 ul high-pressure syringe or sample injection loop
4. Solvent and sample clarification kit (Millipore)
5. Usual laboratory apparatus
-------
Chlorpyrifos EPA-4
(tentative)
Operating Conditions;
Mobile phase: 75% methanol + 25% water
Column temperature: Ambient
Flow rate: 1.5 ml/min
Chart speed: 0.5 in/min (or adjusted)
Amount injected: 10 ul
Conditions may have to be adjusted for the specific instrument
being used, column variations, sample composition, etc. to obtain
optimum response and reproducibility.
Procedure;
Preparation of Standard::
Weigh 0.125 gram chlorpyrifos standard into a 50 ml glass-
stoppered Erlenmeyer flask or 2 oz screw-capped bottle, add by
pipette 25 ml methanol, close tightly and shake to dissolve.
(cone 5 mg chlorpyrifos/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.125 gram chlorpyrifos
into a 50 ml glass-stoppered Erlenmeyer flask or 2 oz screw-capped
bottle, add by pipette 25 ml methanol, shake to extract and dissolve
the chlorpyrifos. Filter through a millipore clarification filter.
(final cone 5 mg chlorpyrifos/ml)
Determination;
Alternately inject three 10 ul portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the percent
chlorpyrifos as follows:
0/ _ (pk. ht. or area sample)(wt. std injected)(?o purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by State of California, Department of Food and Agriculture,
Chemistry Laboratory Services, Sacramento, CA.
Any criticism, suggestions, or data concerning the use of this method
will be appreciated.
-------
March 1978
Deet EPA-4
(tentative)
Determination of Ueet by
High Pressure Liquid Chromatography
(Normal Phase)
Deet is the common name for N,N-diethyl-m-toluamide, a registered
insect repellent having the chemical formula:
CH2-CH3
-CH3
Molecular formula: C H NO
Molecular weight:
Boiling point:
191.3
111°C at 1 mm Hg
Physical state and color: colorless to anber liquid, nearly odorless;
the technical product contains 85-95% m isomer;
the o and p isomers are highly repellent but less
effective than the m isomer
Solubility: practically insoluble in water; miscible with ethanol,
isopropanol, propylene glycol, cottonseed oil, ether,
benzene
Stability: stable under normal conditions; non-corrosive to most metals
Other names: Metadelphene (Hercules), Delphene, Detamide, Off
Reagents;
1. Deet standard of known % purity
2. Chloroform, pesticide or spectro grade
Equipment;
1. High pressure liquid chromatograph with ultraviolet detector
at 254 nm
2. Liquid chromatography column: 30 cm x 4 mm ID, micro Porasil
or equivalent silica column
3. High pressure liquid syringe or sample injection loop
Operating Conditions;
Mobile phase: 60% chloroform + 40% iso-octane
Column temperature: Ambient
Column pressure: 1000 psi (observed)
Flow rate: 1 ml/min
-------
2 Deet EPA-4
(tentative)
Operating Conditions (contM):
Detector: 254 nm
Chart speed: Adjusted
Injection: 5 ul
Conditions may have to be adjusted for the specific instrument
being used, column variations, sample composition, etc. to obtain
optimum response and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.1 gram deel: standard into a 50 ml volumetric flask,
make to volume with chloroform, and mix thoroughly, (cone 2 ug/ul)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.1 gram deet into a
50 ml volumetric flask,, make to volume with chloroform, and mix
thoroughly, (cone 2 ug deet/ul)
Determination;
Using a high pressure liquid syringe or a sample injection
loop, alternately inject three 10 ul portions each of the
standard and sample solutions. Measure the peak height or area
for each peak and calculate the average for both standard and
sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient: size peaks.
Calculation;
From the average peak height or peak area, calculate the
percent deet as follows:
_ (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
Any criticism, data, or suggestions concerning the use of this method
will be appreciated.
-------
September 1978
Diphacinone EPA-2
(tentative)
Determination of Uiphacinone by
High Pressure Liquid Chrotnatography
Using Paired Ion Chromatography
Diphacinone is the accepted common name for 2-(diphenylacetyl)-
1,3-indandione, a registered rodenticide having the chemical structure:
Molecular formula: C^-H.. ,00
2.J ib J
Molecular weight: 340.4
Melting point: 145°C
Physical state, color, and odor: yellow, odorless crystals
Solubility: slightly soluble in water and benzene; soluble in acetone
and acetic acid. Forms a sodium salt which is sparingly
soluble in water.
Stability: resists hydrolysis; stable toward mild oxidants; non-corrosive
Other names: Diphacin (Velsicol Chem. Corp.), diphacin (Turkey), Ramik,
diphenadione
Reagents;
1.
2.
3.
Diphacinone standard of known % purity
Dioxane, ACS
Paired Ion Chromatography (PIC) Reagent A - Add one bottle
of PIC Reagent A to 1000 ml distilled water, stir for 5
minutes, and filter through a 0.45 micron filter.
Each bottle of PIC Reagent A (tetrabutylammonium
phosphate for separation of acids) contains sufficient PIC A
to make one liter of mobile solvent. This solution is fil-
tered through the 0.45 micron filter to remove any suspended
particulate material increasing the useful life of the PIC
solution, and to prevent clogging of the column.
-------
2 Dlphacinone EPA-2
(tentative)
Equipment;
1. High pressure liquid chromatograph with UV detector at 254 nra.
If a variable wavelength UV detector is available, other
wavelengths may be usesd to increase sensitivity or to eliminate
interference (280 nm is very good for diphacinone).
2. 30 cm x 2.0 mm ID Waters C..Q Bondapak or equivalent column
3. High pressure liquid syringe or sample injection loop
4. Usual laboratory glassware
Operating Conditions;
Mobile phase: 90% methanol + 10% aqueous PIC Reagent A
Column temperature: Ambient
Chart speed: 5 miri/inch or equivalent
Flow rate: 0.5 to 1.5 ml/min
Pressure: 1000-1200 psi
Attenuation: Adjusted
Conditions may have to be varied by the analyst for the specific
instrument being used, column variations, sample composition, etc.
to obtain optimum response and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.1 gram standard diphacinone into a 100 ml volumetric
flask, dissolve in, and make to volume with dioxane. Mix thor-
oughly. Pipet 10 ml into a second 100 ml volumetric flask, make
to volume with dioxane, and mix thoroughly. Pipet 10 ml into a
third 100 ml volumetric flask, make to volume with dioxane, and
again mix thoroughly. (cone 10 ug/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.00025 gram
diphacinone (5 grams for a 0.005% formulation) into a small
glass-stoppered flask or screw-cap bottle. Add 25 ml dioxane
and shake on a mechanical shaker for one hour. Allow any solid
matter to settle and filter through a 0.45 micron filter. (cone
10 ug diphacinone/ml)
Determination;
Alternately inject three 10 ul portions each of standard and
sample solutions. Adjustments in attenuation or amount injected
may have to be made to give convenient size peaks.
Measure the peak height or peak area for each peak and
calculate the average for both standard and sample.
-------
Diphacinone EPA-2
(tentative)
Calculation;
From the average peak height or peak area calculate the
percent diphacinone as follows:
7 - (pk« ht. or area sample)(cone, std in ug/ml)(% purity of std)
(pk. ht. or area standard)(cone, sample in ug/ml)
Method submitted by Elmer H. Hayes and Mark VJ. Law, EPA Beltsville
Chemistry Laboratory, Beltsville, Md.
Any criticism, data, or suggestions concerning this method will be
appreciated.
-------
March 1978
Diphenylamine EPA-1
(tentative)
Determination of Diphenylamine
by Gas Liquid Chromatography (TCD)
Diphenylamine is a registered insecticide having the chemical
structure:
Molecular formula: C H -N
Molecular weight: 169.2
Melting point: 53-54°C; boiling point: 302°C
Physical state, color, and odor: white crystalline solid, floral odor
Solubility: insoluble in water; one gram dissolves in 2.2 ml alcohol,
4.5 ml propyl alcohol; freely soluble in benzene, ether,
glacial acetic acid, and carbon disulfide
Stability: discolors in light; forms salts with strong acids
Other names: N-phenylbenzeneamine, Big Dipper, Scaldip
Reagents;
1. Diphenylamine standard of known % purity
2. Benzene, pesticide or spectro grade
Equipment;
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 4' x 1/4" glass column packed with 3.8% SE-30 on
Diatoport S 80/100 mesh (or equivalent column)
3. Precision liquid syringe: 10 ul
4. Usual laboratory glassware
Operating Conditions for TCD;
Column temperature:
Injection temperature:
Detector temperature:
Filament current:
Carrier gas:
Attenuation:
Flow rate:
155°C
200°C
200°C
200 ma
Helium
1
100 ml/min
Operating conditions for filament current, column temperature, or
gas flow should be adjusted by the analyst to obtain optimum response
and reproducibility.
-------
Diphenylamine EPA-1
(tentative)
Procedure:
Preparation of Standard;
Wei?>h 0.15 gram diphenylamine standard into a 25 ml volumetric
flask, make to volume with benzene, and mix thoroughly. (cone
6 ug/ul)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.15 gram diphenyl-
amine into a 25 ml volumetric flask, make to volume with benzene,
and mix thoroughly. (cone 6 ug diphenylamine/ul)
Determination;
Using a precision liquid syringe, alternately inject three
4 ul portions each of standard and sample solutions. Measure the
peak height or peak area for each peak and calculate the average
for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the
percent diphenylamine as follows:
„ _ (pk. ht. or area sample)(wt. std injected)(%_purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Stelios Gerazounis, EPA Pesticide Chemistry Lab-
oratory, Region II, New York, NY.
Any comments, data, or suggestions concerning the use of this method
will be appreciated.
-------
March 1978
Endosulfan EPA-5
(tentative)
Determination of Endosulfan by
Gas Liquid Chromatography
(FID - Internal Standard)
Endosulfan is the accepted common name for Hexachlorohexahydro-
methano-2,4,3-benzodioxathiepin-3-oxide, a registered pesticide
having the chemical structure:
Cl
Molecular formula:
Molecular weight:
Melting point:
CH2—0,
CH2—0'
;s=o
H
406.9
(see below)
Physical state, color, and odor: endosulfan is an odorless white
crystalline solid mixture of two isomers with
mp's of 106°C and 212°C; the technical product
is a brownish crystalline solid, mp 70-100 C,
with a 4:1 ratio of the above isomers.
isomers are insecticidally active.
Both
Solubility:
Stability:
practically insoluble in water, but soluble in most
organic solvents
generally quite stable; decomposition catalyzed by iron;
slowly hydrolyzed by water; sensitive to acid and bases;
compatible with non-alkaline pesticides
Other names: Thiodan (Farwerke Hoechst), Beosit, Chlorthiepin, Cyclodan,
Insectophene, Kop-Thiodan, Malix, Thifor, Thimul, Thionex,
HOE 2671, MIA 5462, FMC 5462
The following method (from N.C. Dept. of Agr.) determines the two
isomers of endosulfan. The ratio of endosulfan I isomer to endosulfan
II isomer in samples ranges from about 4:1 to 2:1. The procedure as
written matches a 2-2/3:1 ratio. Linearity and precision by area
(electronic integration) are very good. Peak height measurements were
not calculated; therefore, should not be used unless a linearity and
precision determination is made. This method is applicable to formula-
tions containing malathion and parathion. Both of these will elute
before endosulfan I and are completely resolved from it.
-------
2 Endosulfan EPA-5
(tentative)
Reagents:
1. Endosulfan I isoraer of known % purity
2. Endosulfan II isomer of known °< purity
3. p-terphenyl, reagent grade
4. Chloroform, pesticide or spectro grade
5. Internal Standard solution - weigh 0.15 gram p-terphenyl
into a 100 ml volumetric flask, make to volume with
chloroform, and mix thoroughly. (cone 1.5 ug/ul)
Equipment;
1. Gas chromatograph with flame ionization detector (FID) and
electronic integrator
2. Column: 6' x 4 mm ID glass column packed with 3% OV-17 on
100/120 mesh Gas Chrom Q (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating^ Conditions for FID;
Column temperature: 205
Injection temperature: 225
Detector temperature: 240
Carrier gas: Nitrogen, 80 cc/min
Carrier gas pressure: 40 psi
Hydrogen pressure: 20 psi
Air pressure: 40 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure;
Preparation of Standard^
Weigh 0.08 gram endosulfan I and 0.03 gram endosulfan II
standards (vary wt. of endosulfan II to match sample more
appropriately if necessary) into a small glass-stoppered flask
or screw-cap bottle. Add by pipet 20 ml of the internal
standard solution and shake to dissolve. (final cone 5.5 ug
total endosulfan and 1.5 ug p-terphenyl/ul)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.11 gram endosulfan
into a small glass-stoppered flask or screw-cap bottle. Add by
pipette 20 ml of the internal standard solution. Close tightly
and shake thoroughly to dissolve and extract the endosulfan.
For coarse or granular materials, shake mechanically for 30
minutes or shake by hand intermittently for one hour. (final
cone 5.5 ug total endosulfan and 1.5 ug p-terphenyl/ul)
-------
3 Endosulfan EPA-5
(tentative)
Determination;
Inject 2 ul of standard and, if necessary, adjust the
instrument parameters to give a complete separation and an
elution time of 6-9 minutes for endosulfan II. The elution
order is endosulfan I, p-terphenyl, then endosulfan II.
Proceed with the determination, making at least three
injections each of standard and sample solutions in random order.
Calculation:
Measure the peak areas of endosulfan I, endosulfan II and
p-terphenyl from both the standard-internal standard solution
and the sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follox^s and calculate the average:
__ , n,. T (peak area endosulfan I)
RF endosulfan I = -)*- -3 •=-= 7T7 1 t: TV
(wt. endosulfan I)(peak area p-terphanyl)
_„ , ,, T_ (peak area endosulfan II)
RF endosulfan 11= •¥-- -5 rr -T\'/ ; : r e rr
(wt. endosulfan II)(peak area p-terphanyl)
(Weights of endosulfan standards should be adjusted according
to % purity.)
Determine the percent endosulfan I and endosulfan II for
each injection of the sample-internal standard solution as
follows and calculate the average:
„, . , .. ,. (peak area endosulfan I) (100)
/, endosulfan I = -?—- ;:—77 ; ; I'\~/V.~T; •: r-z -r-
(wt. sample)(peak area p-terphenyl)(RF endosulfan I)
„ , ^ r TT (peak area endosulfan II)(100)
% endosulfan II = , .— , N > ; *-? r~
(wtTsample) (peak area p-terphenyl) (RF endosulfan II)
% total endosulfan = % endosulfan I -I- % endosulfan II
Method submitted by North Carolina Department of Agriculture,
Pesticide Section, Raleigh, N.C.
Any criticisms, data, or suggestions concerning the use of this method
will be appreciated.
-------
March 1978 EPIC EPA-6
(tentative)
Determination of EPTC
by High Pressure Liquid Chromatography
(Reverse Phase)
EPTC is the common name for S-ethyl dipropylthiocarbamate, a
registered herbicide having the chemical structure:
CH2— CH3
CH3 — CH2 - S — C
— CH2 — CH3
Molecular formula: C H
Molecular weight: 189.3
Boiling point: 127°C at 20 mm Hg (235°C by extrapolation)
Physical state, color, and odor: Light yellow-colored liquid with
an amine odor
Solubility: 365 ppm in water at 20 C; miscible with acetone, benzene,
ethanol, isopropanol, kerosene, methanol, methyl isobutyl
ketone, toluene, and xylene
Stability: stable, non-corrosive
Other names: Eptam (Stauffer), Eradicane, Knoxweed
Reagents;
1. EPTC standard of known % purity
2. Dioxane, pesticide or spectro grade
3. Methanol, pesticide or spectro grade
Equipment;
1. High pressure liquid chromatograph with variable ultraviolet
detector adjustable to 230 nm (254 nm may be used but
sensitivity is less)
2. Liquid chromatographic column, two 1/2 m x 2.0 mm I.D.
Permaphase ODS or equivalent silica column
3. High pressure liquid syringe or 5 ul sample injection loop
Operating Conditions for Perkin-Elmer HPLC;
Mobile phase: 25% methanol + 75% water
Column temperature: Ambient
Column pressure: 2600 psi (observed)
Flow rate: 0.75 ml/min
Detector: Variable wavelength 230 nm
Chart speed: Adjusted
Injection: 5 ul
-------
EPTC EPA-6
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.1 gram EPTC standard into a 50 ml volumetric flask,
make to volume with dioxane, and mix thoroughly, (cone 2 ug/ul)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.1 gram EPTC into a
50 ml volumetric flask. For emulsifiable concentrates make to
volume with dioxane;; for dusts or granules add 50 ml of dioxane
by pipette. Shake thoroughly to dissolve or extract the EPTC.
(cone 2 ug EPTC/ul)
Determination;
Using a high pressure liquid syringe or 5 ul injection loop,
alternately inject three 5 ul portions each of the standard and
sample solutions. Measure the peak height or area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to
be made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the
percent EPTC as follows:
"/ = (pk. ht. or area sample) (wt. std injected) (% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
Any criticism, data, or suggestions concerning the use of this method
will be appreciated.
-------
September 1978
Ethofumesate EPA-1
(tentative)
Determination of Ethofuraesate by
Gas Liquid Chromatography
(FID - Internal Standard)
Ethofumesate is the common name (ISO, BSI, and ANSI pending)
for 2-ethoxy-2,3-dihydro-3,3-dimethyl-5-benzofuranyl methane-
sulphonate, a registered herbicide having the chemical structure:
0 —CH2-CH3
Molecular formula: C _H 0 S
Molecular weight: 286.34
Melting point: 70-72°C
Physical state and color: white, crystalline solid
Solubility: 110 ppm in water; 10% in ethanol; 25% in glycerol;
40% in acetone, benzene, chloroform, and dioxan;
0.4% in hexane
Stability: stable to hydrolysis in water at pH 7
Other names: Nortron' (Fisons Ltd., Great Britain), NC 8438
Note: The name "Nortron" was previously used by Fisons Limited for
"6-chloro-2-trifluoromethy1-3-H-imidazo-(4,5,6) pyridine"
with the proposed ISO name "fluoromidine." This compound
was discontinued and the name "Nortron" was then used for
ethofumesate, the compound described in this method.
-------
2 Ethofumesate EPA-1
(tentative)
Reagents;
1. Ethofumesate standard of known % purity
2. Dipentyl phthalate standard of known % purity
3. Methylene chloride, pesticide or spectro grade
4. Internal Standard solution - weigh 0.75 gram dipentyl
phthalate standard into a 50 ml volumetric flask, dissolve
in, and make to volume with methylene chloride, (cone 15 mg/ml)
Equipment!
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 2 mm ID glass column packed with 5% SE-30 on
Chromosorb W HP 80-100 mesh (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 200 C
Injection temperature: 220°C
Detector temperature: 240 C
Carrier gas: Helium or Nitrogen
Carrier gas flow: 30 ml/min (adjusted for specific GC)
Hydrogen pressure: 20 psi (adjusted for specific GC)
Air pressure: 30 psi (adjusted for specific GC)
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.075 gram ethofumesate standard into a 25 ml volumetric
flask. Add 5 ml of internal standard solution by pipette and
shake to dissolve the ethofumesate. Make to volume with methylene
chloride and mix thoroughly, (final cone 3 mg ethofumesate and
3 mg dipentyl phthalate/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.075 gram ethofumesate
into a 25 ml volumetric flask. Add 5 ml of internal standard by
pipette and shake to dissolve the ethofumesate in the sample. Make
to volume with methylene chloride and mix thoroughly, (final cone
3 mg ethofumesate and 3 mg dipentyl phthalate/ml)
Determination;
Inject 5 ul of the standard-internal standard solution and,
if necessary, adjust the instrument parameters and the volume
injected to give a complete separation within a reasonable time
and to give peak heights of from 1/2 to 3/4 full scale. The
elution order is dipentyl phthalate, then ethofumesate.
-------
Ethofumesate EPA-1
(tentative)
Proceed with the determination, making at least three
injections each of standard-internal standard and sample-
internal standard solutions in random order.
Calculation;
Measure the peak heights or areas of ethofumesate and
dipentyl phthalate from both the standard-internal standard
and the sample-internal standard solutions.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(DPP = dipentyl phthalate = internal standard)
^ = (wt. DPP)(% purity DPP)(pk. ht. or area ethofumesate)
(wt. ethofumesate)(% purity ethofumesate)(pk. ht. or area DPP)
Determine the percent ethofumesate for each injection of
the sample-internal standard solution as follows and calculate
the average:
7 _ (wt. DPP)(% purity DPP)(pk. ht. or area ethofumesate)
(wt. sample)(pk. ht. or area DPP)(RF)
Method submitted by Mark W. Law, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
Any criticism, data, or suggestion concerning the use of this method
will be appreciated.
-------
March 1979
Flammability Test EPA-1
(Flame Projection)
Flame Projection Flammability Test for
Self-Pressurized Aerosol Dispensers
This test indicates the flammability hazard of aerosol formulations
by measuring the length of flame that occurs when an aerosol (self-
pressurized dispenser) is sprayed across a burning candle. Under the
standardized conditions of this test, a flame 18 inches or longer is
considered flammable.
The Flammability Test EPA-2 (Closed Drum) should also be used for
the same aerosol formulation because the two tests together give a
better indication of the flammability hazard than either test alone.
Equipment;
The test equipment consists of a wooden base 8-10 inches wide and
30 inches long. A two-foot scale marked at 1-inch intervals is,mounted
horizontally along one side six inches above the base. The zero point
of the scale starts six inches from the end. A small candle is placed
at this zero point at such a height that the top third of the flame is
even with the scale. The flame should be about 2 inches high.
24'
18'
12" 6'
0
-6'
MI i i i i il i i i i i i i i i i I i i i
Procedure:
The test equipment should be placed in a draft-free area that can
be ventilated to clear the atmosphere after each test.
Place the aerosol at a distance of 6 inches from the flame or at
the end of the test equipment (really minus 6 inches from the ruled
scale). Spray the dispenser so that the spray passes through the top
third of the flame at a right angle to it. Spray for several seconds
while an observer notes the length of flame. The normal bending of
the flame is about 2 inches and is considered part of the flame length.
Take three readings for each aerosol and average the results.
-------
Flammability Test EPA-1
(Flame Projection)
Self-pressurized dispensers should be classed as flammable:
(1) when the length of flame at full value opening is
18 inches or more,
(2) if there is a flash-back to the container at any
degree of valve opening.
-------
March 1979
Flarnmability Test EPA-2
(Closed Drum)
Closed Drum Flammability Test for
Self-Pressurized Aerosol Dispensers
This test indicates the hazard that results from spraying different
aerosol formulations in a closed space in which there is a flame. The
amount of time it takes for a positive result to occur indicates the
hazard from various degrees of dilution with air (longer spraying time
equals higher concentration of the formulation in the atmosphere in
the drum).
The Flammability Test EPA-1 (Flame Projection) should also be used
for the same aerosol formulation because the two tests together give a
better indication of the flammability hazard than either test alone.
Equipment.;
A 55-gallon open-head drum or similar container is fitted with a
hinged (at the top) cover arranged so that it will readily swing open
at a pressure of five pounds. The cover does not have to be "airtight"
but should adequately close the end of the drum.
The opposite end of the drum is equipped with three shuttered
openings—top, side, and bottom—each two inches from the drum's edge
and each one inch in diameter. The end is also fitted with a six-inch-
square observation window covered with safety glass. A side observation
window is optional.
A small candle is placed inside the drum (as it lies on its side) on
the bottom midway between the ends.
L SAMPLE OPENINGS
^VIEWING WINDOWS
-------
Flammability Test EPA-2
(Closed Drum)
Procedure:
The drum should be used out of doors when the temperature is between
60-80 F. If this is not possible, place the drum in a working area that
is properly ventilated.
Open one of the shutters and spray the aerosol (value fully opened)
into the drum for one minute. Clear the atmosphere in the drum and
repeat with each of the other two openings.
Any rapid burning or explosion of the vapor-air mixture sufficient
to cause the hinged cover to move is considered a positive test and is
enough to class the unit being tested as flammable.
-------
March 1978 Fluometuron EPA-2
(tentative)
Determination of Fluometuron
by Ultraviolet Spectroscopy
Fluometuron is the accepted common name for l,l-dimethyl-3-
(a,a,a-trifluoro-m-tolyl) urea, a registered herbicide having the
chemical structure:
Molecular formula: C QH F N 0
Molecular weight: 232.2
Melting point: 163 to 164.5 C (the technical product is about
96% pure and has a m.p. of about 155 C)
Physical state, color, and odor: odorless, white, crystalline solid
Solubility: 90 ppm in water at 25 C; soluble in acetone, ethanol,
isopropanol
Stability: stable, non-corrosive, compatible with other herbicides
Other names: Cotoran (CIBA-Geigy), Lanex (Nor-Am), C-2059, CIBA-2059
Reagents;
1. Fluometuron standard of known % purity
2. Methanol, pesticide or spectro grade
(Other suitable organic solvents such as 95% methanol,
isopropylanol, or chloroform may be used.)
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm silica cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
-------
Fluometuron EPA-2
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.1 gram fluotneturon standard into a 100 ml volumetric
flask. Dissolve in, make to volume with methanol, and mix thor-
oughly. Pipette 10 ml into a second 100 ml volumetric flask,
make to volume with methanol, and mix thoroughly. Pipette 5 ml
of this solution into a third 100 ml volumetric flask, and make
to volume with methanol. Mix thoroughly. (cone 5 ug/ml)
Preparation of Sample;
For wettable powders - weigh a portion of sample equivalent to
0.1 gram fluometuron into a 250 ml glass-stoppered flask or screw-
cap bottle. Add 100 ml methanol, close tightly, and shake for
one hour. Allow to settle, centrifuge or filter if necessary.
Proceed as in the third paragraph beginning "Pipette 10 ml - - -."
For concentrates or high percent formulations (above 90%
fluometuron), weigh a portion of sample equivalent to 0.1 gram
fluometuron into a 100 ml volumetric flask, make to volume with
methanol, and mix thoroughly.
Pipette 10 ml of either of the above sample solutions into a
100 ml volumetric flask, make to volume with methanol, and mix
thoroughly. Pipette 5 ml of this solution into another 100 ml
volumetric flask, make to volume with the methanol, and mix thor-
oughly, (final cone 5 ug fluometuron/ml)
UV Determination;
Balance pen for 0 and 100% transmission at 243 nm with methanol
in each cell. Scan standard and sample from 300 nm to 200 nm with
methanol solution in the reference cell. Measure absorbance of
standard and sample at 243 nm.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent fluometuron as follows;
y - (abs. sample)(cone, std in ug/ml)(% purity std)
(abs. std)(cone, sample in ug/ml)
Method submitted by George Radan, EPA Product Analysis Laboratory,
New York, NY.
Any criticisms, data, or suggestions concerning the use of this method
will be appreciated.
-------
January 1979 Methomyl EPA-1
(tentative)
Determination of Methomyl
by High Pressure Liquid Chromatography
Methomyl is the accepted (ANSI, BSI) common name for S-methyl N-
[_(methylcarbamayl)oxy] thioacetimidate, a registered insecticide and
nematocide having the chemical structure:
0
II
CH3— 'C=N— 0—C—NH—CH3
S-CH3
Molecular formula: CrH,nN?095
Molecular weight: 162.2
Physical state, color, and odor: white crystalline solid with a slight
sulfurous odor
Melting point: 78-79°C
Solubility: solubility at 25 C w/w is 5.8 in water, 73 in acetone,
42 in ethanol, 22 in isopropanol, 100 in methanol,
3 in toluene
Stability: stable in solid form and in aqueous solutions under
normal conditions; subject to decomposition under moist
conditions in soil; aqueous solution is non-corrosive
Other names: Lannate (duPont), Nudrin (Shell)
Reagents;
1. Methomyl standard of known % purity
2. Methanol, spectro or pesticide grade
Equipment;
1. High pressure liquid chromatograph with variable wavelength UV
detector for 233 nm
2. Suitable column such as: Partisil 10 ODS 25 cm x 4.6 mm ID
3. High pressure 10 ul liquid syringe (or suitable sample injection
loop)
4. Mechanical shaker
5. Solvent and sample clarification kit (obtainable from Millipore)
6. Usual laboratory glassware
-------
2 Methomyl EPA-1
(tentative)
Operating Conditions;
Mobile phase: 75% methanol + 25% water
Column temperature: ambient
Flow rate: 1.3-1.5 ml/min
Chart speed: 0.5 cm/min
Conditions may have to be adjusted for the specific instrument
being used, column variations, sample composition, etc. to obtain
optimum response and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.125 gram methomyl standard into a small screw-cap
bottle, add by pipette 25 ml methanol, shake to dissolve and to
mix thoroughly, (final cone 5 mg methomyl/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.125 gram methomyl
into a small screw-cap bottle, add by pipette 25 ml methanol, close
tightly and shake for 15-30 minutes on a mechanical shaker. Allow
to settle and if not clear, centrifuge a portion for a few minutes,
then filter through a millipore filter. Take precaution to prevent
evaporation, (final cone 5 mg methomyl/ml)
Determination:
Alternately inject three 5 ul portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the percent
methomyl as follows:
0/ _ (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt.sample injected)
Method submitted by the Sacramento Pesticide Laboratory, Dept. of Food
and Agriculture, State of California, 1220 N Street, Sacramento, CA 95814.
Any criticism, suggestions, or data concerning the use of this method
will be appreciated.
-------
January 1979
Methoxychlor EPA-3
(tentative)
Determination of Methoxychlor by
High Pressure Liquid Chromatography
(Normal phase)
Methoxychlor, technical is the official name for 2,2-bis (p-methoxy-
phenyl)-l,l,l-trichloroethane 8Q% and related compounds 12%; it is a
registered insecticide having the chemical structure:
CH-7—0
0 — CH3
Molecular formula: C, ,Hn CC1,,0,,
J.D Lj J L.
Molecular weight: 345.5
Physical state, color, and odor: pure p,p' isomer forms colorless crystals;
technical product is a gray flaky powder containing
88?o p,p' isomer with the bulk of the remainder being
the o,p isomer
Melting point: pure p,p" isomer 89 C; technical 70 to 85 C
Solubility: practically insoluble in water; moderately soluble in ethanol
and petroleum oils; readily soluble in most aromatic solvents
Stability: resistant to heat and oxidation; susceptible to dehydrochlor-
ination by alcoholic alkali and heavy metal catalyst
Other names: Marlate (DuPont), Moxie, l,l,l-trichloro-2,2-bis(p-methoxy~
phenyl) ethane
Reagents'.
1. Methoxychlor standard of known % purity
2. Petroleum ether, spectro or pesticide grade
3. Dichloromethane, spectro or pesticide grade
-------
2 Methoxychlor EPA-3
(tentative)
Equipment;
1. High pressure liquid chromatograph with UV detector at 254 nm
2. Suitable column such as: 4 mm ID x 25 cm packed with Partisil
(10 micron)(or equivalent column)
3. 10 ul high pressure liquid syringe or sample injection loop
4. Mechanical shaker
5. Centrifuge
6. Millipore sample clarification kit
7. Usual laboratory glassware
Operating Conditions;
Mobile phase: 80% petroleum ether + 20?o dichloromethane
Column temperature: ambient
Flow rate: 2 ml/min
Chart speed: 0.5 in/min
Amount injected: 10 ul
Conditions may have to be adjusted for the specific instrument being
used, column variations, sample composition, etc. to obtain optimum
response and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.1 gram methoxychlor into a 125 ml glass-stoppered flask
or screw-cap bottle, add (by pipette) 50 ml dichloromethane, dissolve,
and mix thoroughly, (final cone 2 mg methoxychlor/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.1 gram methoxychlor
into a 125 ml glass-stoppered flask or screw-cap bottle, add 50 ml
dichloromethane by pipette, close tightly, and shake for 30 minutes
on a mechanical shaker. Allow to settle; if not clear, centrifuge a
portion a few minutes. Filter a portion through a 5 micron millipore
filter. Take precaution to prevent evaporation, (final cone 2 mg
methoxychlor/ml)
Determination;
Alternately inject three 10 ul portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the percent
methoxychlor as follows:
°' - (pk. ht. or area sample)(wt. std injected)(?° purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
Any criticism, suggestions, or data concerning the use of this method
will be appreciated.
-------
March 1978
Determination of Mexacarbate
by Gas Liquid Chromatography
(TCD - Internal Standard)
Mexacarbate EPA-1
(tentative)
Mexacarbate ±s the accepted common name for A-dimethylamino-3,5-
xylyl methylcarbamate, a registered insecticide and acaricide having
the chemical structure:
—CH3
C12H18N2°2
Molecular formula:
Molecular weight:
Melting point:
Physical state, color, and odor:
Solubility:
222.3
85°C
white, odorless, crystalline solid
Stability:
Other names:
0.01% in water at 25 C; sparingly soluble in petroleum
solvents; very soluble in benzene, ethanol, chloroform,
methylene chloride, acetonitrile, and acetone
decomposed in highly alkaline media; stable under normal
storage conditions; compatible with most other pesticides
Zectran (Dow Chem. Co.), Dowco 139, 4-dimethylamino-3,5-
xylyl N-methylcarbamate, methylcarbamic acid 4-(dimethy1-
amino)-3,5-xylyl ester
Reagents;
1. Mexacarbate standard of known % purity
2. Deet standard of known % purity
3. Acetone, pesticide or spectro grade
4. Internal Standard solution - weigh 7.0 grams deet standard
into a 50 ml volumetric flask, dissolve in and make to volume
with acetone; mix thoroughly. (cone 140 mg deet/ml)
Equipment;
1. Gas chromato.^raph with thermal conductivity detector (TCD)
2. Column: 4' x 1/4" I.D. glass, packed with 3.8% SE-30 on
80/100 Diatoport S or equivalent column (such as
4' x 1/4" I.D. glass, packed with 4% SP-2100 on
80/100 Chromosorb 750)
3. Precision liquid syringe: 10 ul
4. Usual laboratory glassware
-------
2 Mexacarbate EPA-1
(tentative)
Operating Conditions for TCP;
Column temperature: 135 C
Injection temperature: 200°C
Detector temperature: 200 C
Filament current: 200 ma
Carrier gas: Helium
Carrier gas pressure: 40 psi
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Note; Mexacarbate shows a small peak approximately 12 minutes
after injection at above parameters, most possibly due
to the decomposition of carbamate at relatively elevated
temperatures. On standing for several days the small
peak increased significantly, while the main peak
decreased (by about 50%). EPA's New York Chemistry Lab
states: the method given here is rapid enough and the
temperature is relatively low to give reproducible
results (obtained 95-97% of claimed active ingredient).
This method has been used in our lab for two years."
Procedure;
Preparation of Standard;
For emulsifiable concentrates and liquid formulations -
weigh 0.6 gran mexacarbate standard into a 10 ml volumetric
flask; add 5 ml internal standard solution by pipette, make to
volume with acetone, and mix thoroughly. (cone 60 mg mexacarbate
and 70 ng deet/ml)
For dusts, granules, and we_t table powders - weigh 0.6 gram
mexacarbate standard into a small glass-stoppered flask or screw-
cap bottle, add by pipette 5 ml of internal standard solution and
5 ml acetone, close tightly and mix thoroughly. (cone 60 mg
mexacarbate and 70 mg deet/ml)
Preparation of Sample;
For eraulsifiable concentrates and liquid formulations -
weigh a portion of sample equivalent to 0.6 gram mexacarbate into
a 10 ml volumetric flask; add 5 ml of internal standard solution
by pipette, make to volume with acetone, and mix thoroughly.
(final cone 60 mg mexacarbate and 70 mg deet/ml)
For dusts, granules, and wettable powders - weigh a portion of
sample equivalent to 0.6 gram mexacarbate into a small glass-
stoppered flask or screw-cap bottle; add by pipette 5 ml internal
standard and 5 ml of acetone, close tightly and shake on a
mechanical shaker for 10-15 minutes or shake by hand intermittently
for 25-30 minutes. (final cone 60 mg mexacarbate and 70 mg deet/ml)
-------
3 Mexacarbate EPA-1
(tentative)
Determination;
Inject 1-2 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable, time and peak heights of from 1/2
to 3/4 full scale. The elution order is mexacarbate, then deet.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of mexacarbate and deet
from both the standard-internal standard solution and the sample-
internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
„ _ (wt. deet)(% purity deet)(pk.ht. or area mexacarbate)
(wt. mexacarbate)(% purity mexacarbate)(pk. ht. or area deet)
Determine the percent mexacarbate for each injection of the
sample-internal standard solution as follows and calculate the
average:
7 - (wt. deet)(% purity deet)(pk. ht. or area mexacarbate)
(wt. sample)(pk. ht. or area deet)(RF)
Method submitted by George Radan, EPA Product Analysis Lab, Region II,
New York, NY.
Any criticisms, data, or suggestions concerning this method or its use
will be appreciated.
-------
March 1979
Mixed Pesticides EPA-1
(Warfarin & Sulfaquinoxaline)
Determination of Warfarin and Sulfaquinoxaline
in Bait Formulations by HPLC-PIC
Warfarin is the official common name for 3-(alpha-acetonylbenzyl)-
4-hydroxycoumarin, a registered rodenticide having the chemical structure;
Molecular formula:
Molecular weight:
Melting point:
c=o
CH3
C19H16°4
308.3
(dl form) 159 to 161°C
Physical state, color, odor, taste:
Solubility
(dl form) colorless, tasteless,
odorless crystals
practically insoluble in water and benzene, moderately
soluble in alcohols, readily soluble in acetone and
dioxane; forms water-soluble salts with sodium
Stability: stable under normal conditions
Other names: WARF (Wisconsin Alumni Research Foundation), coumafene
(France), zoocoumarin (Netherlands, USSR), Kypfarin
-------
Mixed Pesticides EPA-1
(Warfarin & Sulfaquinoxaline)
Sulfaquirioxaline is the common name for N'-(2-quinoxalinyl) sulfs-
nilamide, a warfarin additive (when added to warfarin formulations it
inhibits the vitamin K producing bacteria in the digestive system of rats
and mice, thereby rendering these rodents more susceptible to the warfarin).
It has the chemical structure:
Molecular formula: C1AH12N4°2S
Molecular weight: 300.33
Melting point: 247-248°C
Physical state, color, and odor:
Solubility:
minute crystals
Stability:
Other names:
solubility in water at pH 7: 0.75 mg/100 ml; in 95?o
alcohol: 73 mg/100 ml; in acetone: 430 mg/100 ml.
Soluble in aqueous Na^CO-, and NaOH solutions
The amorphous salt is deliquescent and absorbs C0? which
liberates the practically insoluble sulfaquinoxaline.
4-amino-N-2-quinoxalinylbenzenesulfonamide; 2-sulfanilamido-
quinoxaline; sulfabenzpyrazine; Compound 3-120; sulquin;
sulfacox; sulfaline; sulfa-Q
Reagents;
1. Warfarin standard of Liown % purity
2. Sulfaquinoxaline standard of known % purity
3. Methanol, pesticide or spectro grade
4. Dioxane, pesticide or spectro grade
5. Water - PIC Reagent A (see note below)
6. Methanol - PIC Reagent A (see note below)
-------
Mixed Pesticides EPA-1
(Warfarin & Sulfaquinoxaline)
Note: Each bottle of PIC (paired ion chromatography)
Reagent A (tetrabutyl ammonium phosphate for
separation of acids) contains sufficient PIC A
to make one liter of mobile solvent. Add one
bottle of PIC A to 1000 ml of water and one
bottle PIC A to 1000 ml of methanol, stir for
about 5 minutes, and filter through a 0.45 micron
filter. These solutions are filtered to remove
any suspended particulate material increasing the
useful life of the PIC solution, and to prevent
clogging of the column.
Equipment;
1. High pressure liquid chromatograph with UU detector at 254 nm.
If a variable wavelength UV detector is available, other wave-
lengths may be useful to increase sensitivity or eliminate
interference. Warfarin is more easily determined at 308 nm.
2. Suitable column such as Waters Bondapak C,g 30 cm x 2.1 mm ID
3. High pressure liquid syringe or sample injection loop
4. Millipore filter apparatus (0.045 micron)
5. Usual laboratory glassware
Operating Conditions;
Mobile phase:
Column temperature:
Chart speed:
Flow rate:
Pressure:
Attenuation:
55?o methanol-PIC A Reagent + 45?o water-PIC A reagent
Ambient
5 min/inch or equivalent
0.5 to .75 ml/min
1000-1400 psi
Adjusted
Conditions may have to be varied by the analyst for the specific
instrument being used, column variations, sample composition, etc. to
obtain optimum response and reproducibility.
Procedure;
Preparation of Standard;
Warfarin - weigh 0.05 gram warfarin standard into a 50 ml
volumetric flask, dissolve in, and make to volume with dioxane,
Mix thoroughly, pipette 5 ml into a second 50 ml volumetric
flask, make to volume with dioxane, and mix thoroughly.
(final cone 0.1 mg warfarin/ml)
Sulfaquinoxaline - weigh 0.05 gram Sulfaquinoxaline standard
into a 50 ml volumetric flask, dissolve in, and make to
volume with dioxane. Mix thoroughly, pipette 5 ml into a
second 50 ml volumetric flask, make to volume with dioxane,
and mix thoroughly, (final cone 0.1 mg sulfaquinoxaline/ml)
-------
4 Mixed Pesticides EPA-1
(Warfarin & Sulf aquinoxaline)
Preparation of Sample ;
Weigh a portion of sample equivalent to 0.005 gram warfarin
(and 0.005 gram sulf aquinoxaline)* into a glass-stoppered or
screw-cap 125 ml Erlenmeyer flask, add 50 ml dioxane by pipette,
close tightly, and shake for one hour. Allow to settle, and
filter through a 0.45 micron millipore filter, (final cone 0.1
gram warfarin and 0.1 gram sulfaquinoxaline/ml)
* Sample and standard weights should be adjusted
as necessary for formulation containing other
than Q.Q25?o of each ingredient.
Determination;
For a variable wavelength detector, use 308 nm rather than
254 nm. Warfarin is more easily detected at this wavelength and
many interferences are eliminated or reduced to a negligible amount.
Alternately inject three 10 ul portions each of standard and
sample solutions. Measure the peak height or peak area for each
peak and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the percent
each of warfarin and sulf aquinoxaline as follows:
ht. or area sample )(wt. std injected)(?o purity of std)
(pk. ht. or area standard) (wt. sample injected)
Method submitted by Elmer H. Hayes, EPA Beltsville Chemistry Laboratory,
Beltsville, MD.
Any criticism, data, or suggestion concerning this method will be
appreciated.
-------
March 1978
Oryzalin EPA-1
(tentative)
Determination of Oryzalin
by Visible (Colorimetric) Spectroscopy
4 4
Oryzalin is the accepted common name for 3,5-dinitro N ,N -
dipropylsulfanilamide, a registered herbicide having the chemical
structure:
CH3—CH2—CH2
CH3—CH2—CH2
Molecular formula:
Molecular weight:
Melting point:
C12H18N4°6S
346.4
141 to 142°C
Physical state, color, and odor: yellow-orange crystalline solid,
no appreciable odor
Solubility: about 2.5 ppm in water at 25 C; readily soluble in polar
organic solvents such as acetone, ethanol, methanol, and
acetonitrile; slightly soluble in benzene and xylene
Stability: susceptible to UV decomposition; non-corrosive; formula-
tions have a shelf life of more than 2 years; technical
material is non-flammable; compatible with most other W.P.
formulations and fertilizers if not highly alkaline
Other names: Surflan and Ryzelan (Eli Lilly & Co.), EL-119, Dirimal
Reagents;
1. Oryzalin standard of known % purity
2. 95% Ethanol, pesticide or spectro grade
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Filtration apparatus
4. Usual laboratory glassware
-------
Oryzalin EPA-1
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.1 gram oryzalin standard into a 100 ml volumetric
flask. Dissolve, make to volume with 95% ethanol, and mix
thoroughly. Pipette a 5 ml aliquot into a 50 ml volumetric
flask, make to volume with 95% ethanol, and again mix thoroughly.
(cone 0.1 mg/ml)
Preparation of Sample:
Weigh a portion of sample equivalent to 0.1 gram of oryzalin
into a 250 ml Erlenmeyer glass-stoppered flask. Add 100 ml 95%
ethanol by pipette and shake on a mechanical shaker for one hour.
Filter if necessary and pipette 10 ml of the clear filtrate into
a 100 ml volumetric flask. Make to volume with 95% ethanol and
mix thoroughly. (final cone 0.1 mg oryzalin/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
(visible range) analytical settings for the particular instrument
being used, balance the pen for 0 and 100% transmission at 385 nm
with 95% ethanol in each cell. Scan both the standard and sample
from 600 nm to 350 nm with 95% ethanol in the reference cell.
Measure the absorbance of both standard and sample at 385 nm.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent oryzalin as follows:
7 - (abs. sample)(cone, std in mg/ml)(% purity std)
(abs. std)(cone, sample in mg/ml)
Method submitted by I. F. Sternman, EPA Product Analysis Laboratory,
New York, NY.
Any criticism, data, or suggestions concerning the use of this method
will be appreciated.
-------
June 1978
Parathion EPA-3
(tentative)
Determination of Parathion in the Presence of Carbaryl
by Gas Liquid Chromatography (FID-IS)
Parathion is the official name for 0,0-diethyl-O-p-nitrophenol
phosphorothioate, a registered insecticide having the chemical
structure:
CH3 - CH2
CH3 - CH2
N02
Molecular formula: CinHi
Molecular weight: 291.3
Melting/Boiling point: m.p
6.0°C, b.p. 157 to 162°C at 6 mm Hg
Physical state, color, and odor: pale yellow liquid; the technical
product is a brown liquid with a garlic-like odor
Solubility:
Stability:
Other names:
Reagents:
1.
2.
3.
4.
24 ppm in water at 25 C; slightly soluble in petroleum oils;
miscible with most organic solvents
rapidly hydrolyzed in alkaline media (at pli 5 to 6, 1% in
62 days at 25°C); isomerizes on heating to the OS-diethyl
isomer
ACC 3422, Thiophos (American Cyanamid); E-605, Folidol
Bladan (Bayer); Niran (Monsanto); Fosferno (Plant Protection
Ltd.); thiophos (USSR); Alkron, Aileron, Aphamite, Corothion,
Ethyl Parathion, Etilon, Fosfono, Orthophos, Pantnion, Para-
mar, Paraphos, Parathene, Parawet, Phoskil, Rhodiatox,
Soprathion, Strathion
Parathion standard of known % purity
Dieldrin standard of known HEOD content
Acetone, pesticide or spectro grade J'elJTiK
Internal standard solution - weigh 0.75 gram^into a 50 ml volu-
metric flask, dissolve in, and make to volume with acetone.
Mix thoroughly. (cone 15 mg/ml)
-------
Parathion EPA-3
(tentative)
Equipment;
1. Gas chroraatograph with flame ionization detector (FID)
2. Column: 4' x 2 mm ID glass column packed with 5% SE-30 on
80/100 riesh Chromosorh W HP (or equivalent column)
3. Precision liquid syringe: 5 or 10 ul
4. Mechanical shaker
5. Centrifuge or filtration equipment
6. Usual laboratory glassware
Operating Conditions for FID;
Column temperature: 175 C
Injection temperature: 250 C
Detector temperature: 250°C
Carrier gas: Helium
Carrier gas pressure: 40 psi (adjusted for particular GC)
Hydrogen pressure: 15 psi (adjusted for particular GC)
Air pressure: 40 psi (adjusted for particular GC)
Chart speed: 0.25"/min or 15"/hr
Operating parameters (above) as well as attenuation and chart
speed should be adjusted by the analyst to obtain optimum response
and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.1 gram parathion into a 50 ml volumetric flask, add
10 ml internal standard solution by pipette, make to volume with
acetone, and mix well. (cone 2 ug parathion and 3 ug dieldrin/ul)
Preparation of Sample;
For liquids and emulsifiable concentrates - weigh a portion of
sample equivalent to 0.1 gram parathion into a 50 ml volumetric
flask, add 10 ml internal standard solution, make to volume with
acetone, and mix well. (cone 2 ug parathion and 3 ug dieldrin/ul)
For dusts and wettable powders - weigh a portion of sample
equivalent to 0.4 gram parathion into a 250 ml glass-stoppered
flask or screw-cap bottle, add 100 ml acetone by pipette. Close
tightly and shake thoroughly to dissolve and extract the parathion.
Shake mechanically for 10-15 minutes or shake by hand intermittently
for 25-30 minutes. Allow to settle, filter or centrifuge if necessary,
taking precaution to avoid loss by evaporation. Pipette a 25 ml
aliquot into a 50 ml volumetric flask, add 10 ml internal standard
by pipette, make to volume with acetone, and mix thoroughly. (final
cone 2 ug parathion and 3 ug dieldrin/ul)
-------
3 Parathion EPA-3
(tentative)
Determination;
Inject 1-2 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from
1/2 to 3/4 full scale. The elution order is parathion, then
dieldrin. (Carbaryl elutes before parathion.) Repeated
injections should give the same peak ratios.
Proceed with the determination, making at least three
injections each of standard and sample solutions in random order,
Calculation;
Measure the peak heights or areas of parathion and dieldrin
from both the standard-internal standard solution and the
sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average;
RF ^ (wt. dieldrin) (% purity dieldrin) (pk. ht. or area parathion)
(wt. parathion) (% purity parathion) (pk. ht. or area dieldrin)
Determine the percent parathion for each injection of the
sample-internal standard solution as follows and calculate the
average:
. dieldrin) (% purity dieldrin) (pk. ht. or area parathion)
(wt. sample) (pk. ht. or area dieldrin) (RF)
Method submitted by the Commonwealth of Virginia, Division of Consol-
idated Laboratory Services, 1 North 14th Street, Richmond, VA 23219.
Checked by Elmer Hayes, EPA Beltsville Chemistry Lab, ARC-East,
Beltsville, MD 20705.
Any criticism, data, or suggestions concerning this method will be
appreciated.
-------
December 1978
Parathion EPA-4
(tentative)
Determination of Parathion in the Presence of Carbaryl
by High Pressure Liquid Chromatography (Reversed Phase)
Parathion is the official name for 0,0-diethyl-O-p-nitrophenol
phosphorothioate, a registered insecticide having the chemical structure;
CH3
Molecular formula: C H^NO^S
Molecular weight: 291.3
Melting/Boiling point: m.p. 6.0°C, b.p. 157 to 162 C at 6 mm Hg
Physical state, color, and odor: pale yellow liquid; the technical
product is a brown liquid with a garlic-like odor
Solubility: 24 ppm in water at 25 C; slightly soluble in petroleum oils;
miscible with most organic solvents
Stability: rapidly hydrolyzed in alkaline media (at pH 5 to 6, 1% in
62 days at 25°C); isomerizes on heating to the QS-diethyl
isomer
Other names: ACC 3422, Thiophos (American Cyanamid); E-605, Folidol
Bladan (Bayer); Niran (Monsanto); Fosferno (Plant Protection
Ltd.); thiophos (USSR); Alkron, Aileron, Aphamite, Corothion,
Ethyl Parathion, Etilon, Fosfono, Orthophos, Panthion, Para-
mar, Paraphos, Parathene, Parawet, Phoskil, Rhodiatox,
Soprathion, 5trathion
Reagents;
1. Parathion standard of known % purity
2. Carbaryl standard of known % purity
3. Dioxane, pesticide or spectro grade
4. Internal standard solution - weigh 3 grams dipropyl phthalate
into a 100 ml volumetric flask, dissolve in and make to volume
with dioxane. Mix thoroughly, (cone 30 mg/ml)
-------
2 Parathion EPA-4
(tentative)
Equipment:
1. High pressure liquid chromatograph with UU detector at 254 nm.
If a variable wavelength UU detector is available, other
wavelengths may be useful to increase sensitivity or to
eliminate interference.
2. Reversed phase column such as DuPont ODS Permaphase, or Perkin
Elmer Sil-X 11 RP
3. High pressure liquid syringe or sample injection loop
4. Millipore filter syringe with 0.45 micron filter pad
5. Usual laboratory glassware
Operating Conditions;
Mobile phase:
Column temperature:
Chart speed:
Flow rate:
Attenuation:
2Q% methanol + 80?o water
50-55°C
5 min/inch or equivalent
0.5 to 1.5 ml/min
Adjusted
Operating parameters (above) as well as attenuation and chart speed
should be adjusted by the analyst to obtain optimum response and repro-
ducibility.
Procedure;
Note: This method is written for a formulation containing a ratio
of 3.5 parts parathion to 1 part carbaryl. If a different
ratio formulation is to be analyzed, use the concentration
of parathion specified (3.5 mg/ml) but change the concen-
tration of carbaryl in the standard to match that in the
sample.
Preparation of Standard;
Weigh 0.35 gram parathion and 0.1 gram carbaryl into a 100 ml
volumetric flask, add 20 ml internal standard solution by pipette,
make to volume with dioxane, and mix thoroughly, (final cone 3.5 mg
parathion, 1 mg carbaryl, and 6 mg dipropyl phthalate/ml)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.35 gram parathion
(and 0.1 gram carbaryl for example) into a 100 ml volumetric flask,
add 20 ml internal standard solution by pipette, and make to volume
with dioxane. Close tightly and place in an ultrasonic bath for
about 5 minutes. Allow to settle or centrifuge a portion and filter
the clear liquid through a 0.45 micron millipore filter, (final
cone 3.5 mg parathion, 1 mg carbaryl, and 6 mg dipropyl phthalate/ml)
-------
Parathion EPA-4
(tentative)
Determination;
Inject 5 ul of standard and, if necessary, adjust the
instrument parameters and the volume injected to give a complete
separation within a reasonable time and peak heights of from 1/2
to 3/4 full scale. The elution order is carbaryl, dipropyl
phthalate, and parathion.
Proceed with the determination, making at least three injec-
tions each of standard and sample solutions in random order.
Calculation;
Measure the peak heights or areas of parathion and dipropyl
phthalate from both the standard-internal standard solution and
the sample-internal standard solution.
Determine the RF value for each injection of the standard-
internal standard solution as follows and calculate the average:
(DPP = dipropyl phthalate)
RF _ (wt. DPP)(?o purity DPP)(pk. ht. or area parathion)
(wt. parathion)(% purity parathion)(pk. ht. or area DPP)
Determine the percent parathion for each injection of the
sample-internal standard solution as follows and calculate the
average:
0, _ (wt. DPP)(?o purity DPP)(pk. ht. or area parathion)
'" = (wt. sample)(pk. ht. or area DPP)(RF)
Calculation of % carbaryl is done in the same way as parathion
using, of course, the carbaryl peaks.
Method submitted by Elmer HI. Hayes, EPA Beltsville Chemistry Laboratory,
Beltsville, MD.
Any criticism, data, or suggestions concerning the use of this method
will be appreciated.
-------
March 1978
Pirimicarb EPA-1
(tentative)
Determination of Pirimicarb in Powder Formulations
by Ultraviolet Spectroscopy
Pirimicarb is the common name for 2-(dimethylamino)-5,6-dimethyl-
4-pyrimidinyl dimethylcarbamate, a registered insecticide (aphicide)
having the chemical structure:
0
0—C—N
\
'CH3
CH3
C11H18N4°2
t
Molecular formula:
Molecular weight:
Melting point:
Physical state, color, and odor:
238
90.5°C
colorless, odorless, crystalline solid
Solubility: 0.27% in water at 25 C; soluble in most organic solvents
Stability: decomposed by ultraviolet light; decomposed by prolonged
boiling with acids or alkalis; forms well-defined water-
soluble crystalline salts with organic and inorganic
acids (HC1 salt is deliquescent); non-corrosive to normal
spray equipment
Other names: Pirimor (Plant Protection Ltd); PP062, 5,6-dimethyl-
2-dimethylamino-4-pyrimidinyl dimethylcarbamate;
2-dimethylamino-5,6-dimethylpyrimidin-4-yl dimethylcarbamate
Reagents;
1. Pirimicarb standard of known % purity
2. Chloroform, pesticide or spectro grade
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Filtration apparatus
4. Usual laboratory glassware
-------
Pirimicarb EPA-1
(tentative)
Procedure:
Preparation of Standard:
Weigh 0.1 gram pirimicarb standard into a 100 ml volumetric
flask. Dissolve, make to volume with chloroform, and mix thor-
oughly. Pipette a 10 rd aliquot into a 50 ml volumetric flask
and make to volume with chloroform. Mix thoroughly and pipette
a 10 ril aliquot into a 50 ml volumetric flask. Make to volune
and again mix thoroughly. (final cone 40 ug/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.1 grari of pirimicarb
into a 250 ml Krlenmeyer glass-stoppered flask. Add 100 ml chloro-
form by pipette, close tightly, and shake on a mechanical shaker
for one hour. Filter if necessary (taking precaution to avoid
loss by evaporation), and pipette 10 ml of the clear filtrate
into a 50 ml volumetric flask. Make to volume with chloroform,
mix thoroughly, and nipette 10 ml into a 50 ml volumetric flask.
Make to volume with chloroform and mix thoroughly. (final cone
40 ug pirimicarb/ml)
UV Determination;
With the UV spec trophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 307 nn with
chloroform in each cell. Scan both the standard and sample from
360 nm to 220 nm with chloroform in the reference cell. Measure
the absorbance of both standard and sample at 307 nm.
(absorbance was Eound to be linear at least to 45 ug/ml)
Calculation;
From the above ahsorbances and using the standard and sample
concentrations, calculate the percent pirimicarb as follows:
7 = (abs- sample) (cone, std in ug/ml) (% purity std)
(abs. std) (cone, sample in ug/ml)
Method submitted by Stelios Gerazounis, EPA Product Analysis Lab-
oratory, New York, NY.
Any criticism, data, or suggestions concerning the use of this method
will be appreciated.
-------
March 1978
Pyrazon EPA-1
(tentative)
Determination of Pyrazon in Powder Formulations
by Ultraviolet Spectroscopy
Pyrazon is the accepted common name for 5-amino-4-chloro-2-
phenyl-3(2H)-pyridazinone, a registered herbicide having the
chemical structure:
C)
Molecular formula:
Molecular weight:
Melting point:
C1()H8C1N30
221.6
pure 205-206°C (207° with decomposition);
technical 185-195°C
tan to brown powder; odorless
Physical state, color, and odor:
when pure
Solubility: 400 ppm in water at 20°C; 2.8% in acetone; 2.4% in
methanol; 0.07% in benzene and in ether; 0.21% in
chloroform; 0.6% in ethyl acetate
Stability: decomposes at melting point; relatively resistant to
decomposition by UV; non-corrosive; nonflammable; no
limitation on shelf life of concentrate
Other names: Pyramin (BASF Canada), PCA, H119
Reagents;
1. Pyrazon standard of known % purity
2. Methanol, pesticide or spectro grade
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm silica cells
2. Mechanical shaker
3. Filtration apparatus
4. Usual laboratory glassware
-------
Pyrazon EPA-1
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.1 gram pyrazon standard into a 100 ml volumetric
flask. Dissolve in, make to volume with methanol, and mix
thoroughly. Pipette a 5 ml aliquot into a second 50 ml volu-
metric flask and make to volume with methanol. Mix thoroughly
and pipette a 5 ml aliquot into a third 50 ml volumetric flask.
Make to volume and again mix thoroughly, (final cone 10 ug/ml)
Preparation of Sample;
Weigh a portion of sample equivalent to 0.1 gram of pyrazon
into a 250 ml Erlenmeyer glass-stoppered flask. Add 100 ml
methanol by pipette and shake on a mechanical shaker for one
hour. Filter if necessary and pipette 5 ml of the clear filtrate
into a 50 ml volumetric flask. Make to volume with methanol, mix
thoroughly and pipette 5 ml into a second 50 ml volumetric flask.
Make to volume with methanol and mix thoroughly. (final cone
10 ug pyrazon/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative
analytical settings for the particular instrument being used,
balance the pen for 0 and 100% transmission at 286 nm with
methanol in each cell. Scan both the standard and sample from
360 nm to 220 nm with methanol in the reference cell. Measure
the absorbance of both standard and sample at 286 nm.
Calculation;
From the above absorbances and using the standard and sample
concentrations, calculate the percent pyrazon as follows:
y _ (abs. sample)(cone, std in ug/ml)(% purity std)
(abs. std)(cone, sample in ug/ml)
Note; It has been established that there is a straight line
relationship between absorbance and concentration.
Method submitted by Stelios Gerazounis, EPA Pesticides Chemistry
Laboratory, Region II, New York, NY.
Any comments, criticism, suggestions, data, etc. concerning the use
of this method will be appreciated.
-------
March 1978
Strychnine EPA-3
(tentative)
Determination of Strychnine
by High Pressure Liquid Chromatography
(Reverse Phase)
Strychnine is a registered rodenticide having the chemical
structure:
0-CH2
Molecular formula:
Molecular weight:
Melting point:
C21H22N2°2
334.4
268 to 290°C (depending on the speed of heating)
with decomposition; b.p. 270 C at 5 mm
Physical state, color, and odor: hard white crystals or powder, very
bitter taste; very poisonous!
practically insoluble in water, alcohol, ether; slightly
soluble in benzene, chloroform
Solubility:
Stability: forms salts with acids; ppt. by alkaloid precipitants
Other names: Kwik-kil, Mouse-tox, Ro-Dec
Reagents;
1. Strychnine standard of known purity
2. Dioxane, pesticide or spectre grade
3. Methanol, pesticide or spectro grade
Equipment;
1. High pressure liquid chromatograph with UV detector at 254 nm.
If a variable wavelength UV detector is available, other wave-
lengths may be useful to increase sensitivity or eliminate
interference.
2. Column - DuPont ETH Permaphase, 1/2 m x 2.1 mm I.D.
-------
2 Strychnine EPA-3
(tentative)
Equipment (cont'd):
3. High pressure liquid syringe or sample injection loop
4. Sample grinder or pulverizer
5. Mechanical shaker
6. 5 micron millipore filter
7. Usual laboratory glassware
Operating Conditions;
Mobile phase: 97-98% HO + 3-2% MeOH (percents may be
varied to obtain optimum separation)
Column temperature: 40 C
Chart speed: 12"/hr
Conditions may have to be varied by the analyst for the specific
instrument being used to obtain optimum response and reproducibility.
Procedure;
Preparation of Standard;
Weigh 0.1 gram strychnine standard into a 100 ml volumetric
flask and make to volume with dioxane. Place into ultrasonic
bath for a few minutes to hasten solution. (cone 1 ug/ul)
Preparation of Sample;
Grind 20-25 grams of sample to a fairly fine state in a
suitable sample grinder., Weigh an amount of sample equivalent
to 0.1 gram strychnine into a 250 ml Erlenmeyer glass-stoppered
flask or small screw-cap bottle. Add 100 ml dioxane by pipet,
close tightly, and shake on a mechanical shaker for 2 hours.
Allow to settle, and filter a portion through a 5 micron milli-
pore filter. (cone 1 ug strychnine/ul)
Determinat ion;
Using a high pressure syringe or sample injection loop,
alternately inject 5 ul portions each of standard and sample
solutions. Measure the peak height or peak area for each peak
and calculate the average for both standard and sample. Adjust-
ments in attenuation or amount injected may have to be made to
give convenient size peaks.
Calculation:
From the average peak height or peak area calculate the percent
strychnine as follows:
7 = (pk. ht. or area sample)(wt. std injected)(% purity of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
Any criticism, data, or suggestions concerning the use of this method
will be appreciated.
-------
March 1978 Vernolate EPA-4
(tentative)
Determination of Vernolate
by High Pressure Liquid Chromatography
(Reverse Phase)
Vernolate is the common name for S-propyl dipropylthiocarbamate,
a registered herbicide having the chemical structure:
CH3 - CH2 — CH2 —
Molecular formula: C10H21NOS
Molecular weight: 203.4
Boiling point: 140°C at 20 mm Hg, 150°C at 30 mm Hg
Physical state, color, and odor: clear liquid with an aromatic odor
Solubility: about 100 ppm in water at 20-21°C; miscible with common
organic solvents
Stability: stable; non-corrosive
Other names: Vernam (Stauffer); R-1607; S-propyl N,N-dipropyl thio-
carbamate
Reagents;
1. Vernolate standard of known % purity
2. Dioxane, pesticide or spectro grade
3. Acetonitrile, pesticide or spectro grade
Equipment ;
1. High pressure liquid chromatograph with variable ultraviolet
detector. Greatest sensitivity is obtained at 230 nm; however,
254 nm can be used with a more concentrated sample.
2. Liquid chromatographic column, one meter x 2.0 mm I.D.
Permaphase ODS or equivalent silica column
3. High pressure liquid syringe or sample injection loop
4. Usual laboratory glassware
Operating Conditions for Perkin-Elmer HPLC;
Mobile phase: 75% water + 25% acetonitrile
Column temperature: Ambient
Column pressure: 1800 psi (observed)
Flow rate: 0.75 ml/min
Detector: Variable at 230 nm for greater sensitivity;
fixed 254 nm can be used if sample is concentrated
Chart speed: Adjusted
Injection: 5 ul
-------
Vernolate EPA-4
(tentative)
Procedure:
Preparation of Standard;
Weigh 0.1 gram vernolate standard into a 50 ml volumetric
flask, make to volume with dioxane, and mix thoroughly.
(cone 2 ug/ul)
Preparation of Sample;
Weigh an amount of sample equivalent to 0.1 gram vernolate
into a 50 ml volumetric flask. For emulsifiable concentrates
make to volume with dioxane; for dusts or granules add 50 ml
of dioxane by pipette. Shake thoroughly to dissolve or extract
the vernolate. (cone 2 ug vernolate/ul)
Determination;
Using a high pressure liquid syringe or 10 ul injection loop,
alternately inject three 10 ul portions each of the standard and
sample solutions. Measure the peak height or area for each peak
and calculate the average for both standard and sample.
Adjustments in attenuation or amount injected may have to be
made to give convenient size peaks.
Calculation;
From the average peak height or peak area calculate the
percent vernolate as follows;
j = (pk. ht._ or area sample) (wt. std injected) (%_purjLtv of std)
(pk. ht. or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA Beltsville Chemistry Laboratory,
Beltsville, Md.
Any criticism, data, or suggestions concerning the use of this method
will be appreciated.
-------
May 1978 TLC Identification EPA-2
Detection of Organothiophosphates
by Thin Layer Chromatography
Organothiophosphates in pesticide formulations can be identified
by spotting (directly for liquids or an ether-hexane extract for dusts
and granules) on precoated plastic sheets and developing in benzene.
The separated spots are color-developed by spraying with 2,6-dibromo-
N-chloro-p-benzoquinoneimine in cyclohexane and exposing to hydro-
chloric acid fumes giving a red to orange-brown color which is char-
acteristic of the particular organothiophosphate present.
Reagents;
1. Benzene, pesticide grade
2. 1:1 diethyl ether-hexane mixture, pesticide grade
3. 2,6-dibromo-N-chloro-p-benzoquinoneimine, 0.5% solution in
cyclohexane. This reagent and its solutions should be kept
refrigerated.
Notel The above chemical is a suspected carcinogen
and should be handled accordingly.
4. Concentrated hydrochloric acid
Equipment;
1. Precoated plastic sheets for TLC, MN Polygram Sil G (0.25 mm
silica gel without gypsum) , available from Brinkman Instru-
ments Inc.
2. Spotting template
3. Spotting pipettes, 1 to 10 ul, or 10 ul GC syringe
4. Two airtight developing tanks: one for benzene and one for
hydrochloric acid
5. Sprayer
Safety note; When toxic or highly corrosive reagents are sprayed
on chromatograms, it is necessary to use gloves,
face shield, respiratory mask, and appropriate fume
hood to protect skin, eyes, and respiratory tract
against mists or fumes generated by the spraying
device.
Procedure;
Preparation of Sample;
For dusts or granules, shake approximately 2 grams with 10 ml
of 1:1 ether-hexane mixture in a 25 ml screw-cap test tube for 10
minutes. Allow to settle, centrifuge if necessary.
liquids, use sample directly.
Spotting;
With a spotting template for a guide, mark the TLC sheet for
sample and standard spots and mark a line at 10 cm using a soft
lead pencil.
-------
TLC Identification EPA-2
Spot 10 ul of sample extract or 2 ul of undiluted liquid samples.
Appropriate standards should be spotted (1 to 5 ug) among the samples.
Ghromatogram and Color Development;
Develop the TLC sheet in benzene in a closed airtight developing
tank until the benzene reaches the 10 cm line. Evaporate all the
benzene from the plate in a hood. Spray with 0.5% 2,6-dibromo-\-
chloro-p-benzoquinoneimine in cyclohexane and allow to evaporate.
Place sheet in a developing tank containing concentrated hydro-
chloric acid fumes, close tightly and leave for about one minute.
Organothiophosphates will start appearing in 10 to 15 seconds
as red to orange-brown spots, the colors being characteristic of the
particular organothiophosphates present. Any spots that appear
before exposing the sheet to the acid fumes should be disregarded.
Detection Limit and Rf Values;
The lower limit of detection of most organothiophosphates is
about 0.25 ug. This allows the detection of a contamination of
approximately 0.01%.
The Rf values of some organothiophosphates using benzene are as
follows:
dimethoate (Cygon) .00
demeton methyl (Hetasystox) .00
demeton (Systox) .00 and .55
azinphos-methyl (Guthion) methyl .07
aeinphos-ethyl (Guthion) ethyl .07
diazinon .15
phosmet (Imidan) streak to .15
malathion .20
coumaphos (Co-Ral) .30
fenthion .48
methyl parathion .50
ethion .53
sulfotepp .55
oxydisulfoton (Disyston-S) .57
parathion .57
dicapthon .57
EPN .58
disulfoton (Disyston) .60
sulfallate (Vegadex) .60
phorate (Thimet) .66
(Aspon) .66
ronnel .70
carbophenothion (Trithion) .74
chlorpyrifos (Dursban) .75
(DBF) . 80
Names in ( ) are trade names.
-------
TLC Identification EPA-2
Method prepared and submitted by Danny D. McDaniel and Robert Robertson,
EPA Pesticide Products Analysis Lab, National Space Technology Lab-
oratories, Bay St. Louis, MS 39529.
This is the method used in the above laboratories for TLC identifi-
cation of organothiophosphates.
It is based on the following references:
Menn, J. J., Erwin, W. R. and Gordon, H. T., J. Agric. Food Chem.,
5, 601 (1957)
Braithwaite, D. P., Nature, 200, 1011 (1963)
Getz, Melvin E., J.A.O.A.C., Vol. 45, No. 2, 393-396 (1962)
Bontoyan, Warren, J.A.O.A.C., Vol. 49, No. 6, 1169-1174 (1966)
-------
-------
Preface
Enclosed is the third update to the EPA Manual of Chemical Methods for
Pesticides and Devices. This update includes 55 new methods. Also included
is a list of these 55 methods and a Pesticide Name Cross Reference Index to
the 55 Methods.
Continuing with the aim of providing suitable methods that can be used to
support enforcement actions, we will appreciate receiving new methods for
inclusion in future updates or revisions of this manual. However, in order to
limit the ever-expanding size of this manual, we also would appreciate your
telling us which specific methods you think should be studied collaboratively
for inclusion in the AOAC Official Methods of Analysis.
For the next update, any suggestions for additional methods, graphs, charts,
data, or information (general or specific) will be appreciated now or at any
future time. Any and all ideas to make this manual more useful are welcome.
Such comments may be made to the Editors.
Editors: Warren R. Bontoyan
Jack B. Looker
Chemical and Biological
Investigations Branch
Environmental Protection Agency
Building 402, ARC-East
Beltsville, MD 20705
-------
-------
Third Update
Pesticide Name Cross Reference Index to the Methods
(55 methods - August)
3336 Thiophanate EPA-1
AAtrex - -Atrazine EPA-3 5 EPA-4
AC 92553 Pendimethal in EPA-1
Acaron Chlordimeform EPA-1
Accotab Pendimethal in EPA-1
3-(aIpha-acetonylbenzyl)-k-
hydroxycoumarin Warfarin EPA-4
ACP 322 - Naptalam EPA-1
Alanap Naptalam EPA-1
Altos id Methoprene EPA-1
Altosid Briquets Methoprene EPA-1
Antene Ziram EPA-1
Antimicrobial Pentachlorophenol EPA-1 & EPA-2
Antu EPA-1 UV
Apprex Tetrachlorovinphos EPA-1
Aquae i de Diquat EPA-1
Atranex Atrazine EPA-3 & EPA-A
Atratol Atrazine EPA-3 & EPA-4
Atrazine EPA-3 - HPLC
Atrazine EPA-fr HPLC (IS)
atrazine (with metolachlor) Mixed pesticides EPA-2
barbasco Rotenone EPA-2
Bay 276 Disulfoton EPA-2
Bay 19639 Disulfoton EPA-2
Bay 251^1 fensulfothion
Bayer 15922- Trichlorfon EPA-3 & EPA-4
Bendiocarb EPA-3 - HPLC (I S)
1,2-benzenedicarboxy1ic acid
dimethyl ester Dimethyl phthalate EPA-1
Bermat Chlordimef orm EPA-1
Bicep metolachlor
-------
Bidr in Dicrotophos EPA-1
1,2-bis(3-ethoxycarbony1-2-
thioureido) benzene • Thiop'nanate EPA-1
Black Leaf 40 Nicotine EPA-1
Bovinox — - Trichlorfon EPA-3 & EPA-4
Briten - - -Trichlorfon EPA-3 & EPA-4
3-tert-butyl-5-chloro-methyluraci 1 Terbaci / EPA-1
1-n-butyl-3-(3,4-dichlorophenyl)-1-
methyl urea Neburon EPA-2
1-(5-tert-butyl-1,3,4-thiadiazol-
2-yl )-1 ,3-dimethyl urea • Tebuthiuron EPA-1
C 709-- -- Dicrotophos EPA-1
C 8514 - Chlordimeform EPA-1
Caid Chiorophacinone EPA-2 & EPA-3
Cal math ion Ma lath ion EPA-3
Captan EPA-3- GC-FID-IS
Captan EPA-4- - HPLC (IS)
captane (France) Captan EPA-3 & EPA-4
Carbarnine Carbaryl EPA-3
Carbaryl EPA-3- -HPLC (IS)
Carbazinc • • Zi ram EPA-1
Carbicron • Dicrotophos EPA-1
carbofos (USSR) - Malathion EPA-3
Cekubaryl Carbaryl EPA-3
Cekufon— Trichlorfon EPA-3 & EPA-1*
Cekumethion • Methyl pa rath ion EPA-6
Celmone Naphtha 1 eneacetic acid EPA-1
Celthion - - Malathion EPA-3
Cercobin Thiophanate EPA-1
Cerobin-M • Thiophanate-methyl EPA-1
CF 125 Chloroflurecol-methyl ester EPA-1
CGA 24705" - metolachlor
Chem Fish Rotenone EPA-2
ChemStorr Prop ionic acid EPA-1
-------
3
Chlorfenamidine (former name) Chi ordimeform EPA-1
chlorinated dibenzo-p-dioxins Dioxins EPA-1
Chlordimeform EPA-1- -GC-FID-IS
2-chloro-A-ethylamono-6-
isopropylamino-1,3,5-triazine -Atrazine EPA-3 & EPA-4
6-chloro-N-ethyl-N'-0-methylethyl)-
1,3,5-triazine-2,4-diamine Atrazine EPA-3 & EPA-4
chlorof lurenol-methyl ester -Chi orof lurecol-methyl ester EPA-1
Chloroflurecol-methyl ester EPA-1 UV
chlorofos-- Trichlorfon EPA-3 & EPA-4
chloromethlyoxirane Epichlorohydrin EPA-1
Chloroneb EPA-1- -UV
Chlorophacinone EPA-2 HPLC
Chlorophacinone EPA-3 _UV
2 [(p-chlorophenyl)phenylacetyl]-
1 ,3-indandione Chlorophacinone EPA-2 & EPA-3
chloropropyl ene oxide Epi chlorohydr in EPA-1
N'-(4-chloro-o-tolyl)-N,N-
dimethyl formamidine Chlordimeform EPA-1
2-chloro-1-(2,4,5-trichloropheny1)
vinyl dimethyl phosphate, cis isomer Tetrachlorovinphos EPA-1
2-chloro-1-(2,4,5-trichlorophenyl
vinyl dimethyl phosphate, I isomer --Tetrachlorovinphos EPA-1
Ciclosom —Trichlorfon EPA-3 & EPA-4
Cobex Din itramine EPA-1
Cobexo Din itramine EPA-1
Codal metol achlor
Co-Rax Warfar in EPA-4
Corozate Zi ram EPA-1
Cotoran Mult i metol achlor
coumafene (France) Warfarin EPA-4
Cov-R-Tox Warfar in EPA-^
Crinex-- Trichlorfon EPA-3 & EPA-4
Crisatrina Atrazine EPA-3 & EPA-A
Crisazine Atrazine EPA-3 & EPA-A
-------
Crisquat Paraquat EPA-1
cube1 Rotenone EPA-2
Cuman Zi ram EPA-1
Curb?set Chlorof lurecol -methyl ester EPA-1
CVMP - Tetrachlorvinphos EPA-1
Cynoff - Pendimethal in EPA-1
Cythion Ma lath ion EPA-3
Danex - - Trichlorfon EPA-3 & EPA-4
Dasanit fensulfothion
deiquat (Germany) Diquat EPA-1
Demosan Chloroneb EPA-1
Denapon Carbaryl EPA-3
derr is Rotenone EPA-2
Detmol MA 36% (Albert & Co. Germany) Malathion EPA-3
Devicarb Carbaryl EPA-3
Devi th ion Methyl pa rath ion EPA-6
Dextrone Diquat EPA-1
Dextrone X — Paraquat EPA-1
Dexuron Paraquat EPA-1
dibenzo-p-dioxin Dioxins EPA-1
Dicarbarn Carbaryl EPA-3
1,4-dichloro-3,5-dimethoxybenzene ---Chloroneb EPA-1
2-(3,^-dichlorophenyl)-A-methyl"1,2,4-
oxadiazol idine-3,5-dione Methazole EPA-1
Dicrotophos EPA-1 _j_R
S-[1,2-di(ethoxycarbonyl)-ethyl]
dimethyl phosphorothiolothionate Malathion EPA-3
0,0-diethyl S- 2-(ethylthio)ethyl
phosphorodi thioate • Disulfoton EPA-3
0,0-diethyl) [4-(methyIsulfinyl)phenyl]
phosphor odi thioate fensulfothion
diethyl-p-methyIsulf inylphenyl
thiophosphate f ensulfoth ion
diethyl [1,2-phenylene bis (imino-
carbonothioyl) ] bis [carbamate] Thiophanate EPA-1
-------
-Thiophanate EPA-1
diethyl ^A-o-phenyl enebi s 13-
th ioallophanate]
N\N^-diethyl-alpha,alpha, alpha-
tr ifluoro-3,5-din itrotoluene-
2,4-diamine Dini tramine EPA-1
S-(2,3-dihydro-5-methoxy-2-oxo-1,3,^-
thiadiazol-3-ylmethyl) dimethyl
phosphorothiolothionate Methidathion EPA-2
6,7-dihydropyrido [1,2-a:2',1'-c]
pyrazinedin ium ion Diquat EPA-1
3-(dimethoxyphosph inyloxy)-N,N-
dimethyl- cis-crotonami de Dicrotophos EPA-1
dimethyl 1,2-benzenedicarboxyIate Dimethyl phthalate EPA-1
2,2-dimethy]-1,3-benzodioxol -*f-y]
N-methylcarbamate Bendiocarb EPA-3
1,1 '-dimethyl -k,k '-bipyridyl ium ion Paraquat EPA-1
0,0,-dimethyl S-(1,3-dicarbethoxyethyl)
phosphorodi thioate- Ma lath ion EPA-3
N,N-dimethyl-2,2-diphenylacetamide Diphenamid EPA-1 & EPA-2
0,0-dimethyl dithiophosphate of
diethyl mercaptosuccinate Malathion EPA-3
N-[5-(1,1-dimethyl ethyl)-1,3,A-
thiadiazol-2-yl]-N,N-dimethyl urea Tebuthirun EPA-1
N,N-dimethyl-N'_(2-methyl-4-chlorophenyl)-
formamidine Chlordimeform EPA-1
0,0-dimethyI-0-p-nitropheny1
phosphorothioate Methyl parathion EPA-6
Dimethyl parathion Methyl parathion EPA-6
dimethyl [(1,2-phenylene) bis-
iminocarbonylthioyl) ] bis
[carbamate] Thiophanate-methyl EPA-1
0,0-dimethyl phosphorodithioate
S-ester with ^-(mercaptomethyl)-2-
methoxy-delta 2-1,3,A-
thiadiazol in-5-one Methidathion EPA-2
Dimethyl phthalate EPA-1- -GC-FID-IS
dimethyl (2,2,2-trichloro-1-
hydroxyethyl) phosphonate Trichlorfon EPA-3 & EPA-4
-------
Dinitramine EPA-1 - -GC-F1D-IS
Dioxins EPA-1- -GC/HS
Diphacin Diphacinone EPA-3
diphacin (Turkey) Diphacinone EPA-3
Diphacine Meal Bait Diphacinone EPA-3
Diphacinone EPA-3 HPLC
Diphenamid EPA-1 _[R_
Diphenamid EPA-2- - • GC-FID-IS
2-(diphenyl acetyl )-1,3-indanedione Diphacinone EPA-3
Dipterex Trichlorfon EPA-3 & EPA-1*
Diguat EPA-1- HPLC (IS)
disul foton Mixed Pesticides EPA-3
Disulfoton - GC-FID-IS
Disyston (with fensulfothion) Disulfoton EPA-3
Di-Syston (in U.S.) Disulfoton EPA-3
dith iodemeton Disulfoton EPA-3
dithiosustox -- Disulfoton EPA-3
DMP Dimethyl phthalate EPA-1
Dowicide Pentachlorophenol EPA-1 & EPA-2
Dowicide EC-7 Pentachlorophenol EPA-1 & EPA-2
Dowicide G Pentachlorophenol EPA-1 & EPA-2
Dowlap Lamprecid (Trade Name) EPA-1
Dow Pentachlorophenol DP-2 Pentachlorophenol EPA-1 & EPA-2
Drat Chlorophacinone EPA-2 & EPA-3
Drexel Methyl Parathion *tE Methyl parathion EPA-6
Drupina 90 Ziram EPA-1
DuPont Herbicide 732 Terbaci 1 EPA-1
Dust M Tetrachlorvinphos EPA-1
Dyanap - Naptalam EPA-1
Dyfonate Fonofos EPA-1
Dylox Trichlorfon EPA-3 6 EPA-^f
Dymid Diphenamid EPA-1 & EPA-2
-------
E 601 Methyl parathion EPA-6
Ektafos Dicrotophos EPA-1
EL 103 - - Tebuthiuron EPA-1
Emmatos Ma lath ion EPA-3
Emmatos Extra Malathion EPA-3
Enide-- Diphenamid EPA-1 & EPA-2
Epichlorohydrin EPA-1 GC-FID
Equino-Aid Trichlorfon EPA-3 & EPA-A
Esgram Paraquat EPA-1
1,1 '-ethylene-2,2'-bipyr idyl ium ion Diquat EPA-1
0-ethyl S-phenyl ethlyphosphonodithioate Fonofos EPA-1
N-(l-ethylpropyl)-3,^-dimethyl-2,6-
din itrobenzeneamine Pendimethal in EPA-1
N-(1-ethyl propy l)-2,6-d in itro-3,1*-
xyl idine-- Pendimethal i n EPA-1
f ensulfoth ion (with disulfoton) Mixed pesticides EPA-3
Ficam Bendiocarb EPA-3
Folidol M Methyl parathion EPA-6
Folosan Tecnazene EPA-1
Fonofos EPA-1- -JJ*
ForMal Malathion EPA-3
Fosferno M 50 Methyl parathion EPA-6
Fruitone N • Naphthaleneacetic acid EPA-1
Frumin AL--- Disulfoton EPA-3
Fuclasin Ultra --Ziram EPA-1
Fuklas in Ziram EPA-1
Fundal • •—Chlordimeform EPA-1
Fund ex Chlordimeform EPA-1
Fungi tox Thiophanate-methyl EPA-1
Fungostop Zi ram EPA-1
Fusarex Tecnazene EPA-1
Fyfanon Malathion EPA-3
-------
G 30027 Atrazine EPA-3
Galecron Chlordimeform EPA-1
Gardeide Tetrachlorvinphos EPA-1
Gar dona Tetrachlorvinphos EPA-1
Garvox Bend iocarb EPA-3
Gearphos Methyl pa rath ion EPA-6
Gesaprim Atrazine EPA-3 & EPA-4
Go-Go-San Pendimethal in EPA-1
Grain Treat Propionic acid EPA-1
Gramonol Paraquat EPA-1
Gramoxone Paraquat EPA-1
Gramuron Paraquat EPA-1
Granurex Neburon EPA-2
Graslan Tebuthiuron EPA-1
Griffex - • Atrazine EPA-3 £ EPA-4
GS 13005-- - Methidathion EPA-2
haiari -- Rotenone EPA-2
Herbadox Pendimethal in EPA-1
Herboxone Paraquat EPA-1
Hexachlorophene EPA-1 HPLC
Hexavin Carbaryl EPA-3
Hexazii Zi ram EPA-1
Hex ide Hexachlorophene EPA-1
Hi Ithion Malathion EPA-3
Isobac (sodium salt) Hexachlorophene EPA-1
2,3-isopropylidened ioxyphenyl
methyl carbamate Bend i oca rb EPA-3
isopropyl (2E, ^E)-11-methoxy-3,7,11-
trimethyl-2,^-dodecodienoate Methoprene EPA-1
IT 3456 Chlorof lurecol -methyl ester EPA-1
-------
Karbaspray --- ...... - ...... -- ...... -- ........ Carbaryl EPA-3
Karbofos-- ................ -- ..... ----------- Ma lath ion EPA-3
Kloben ------- ...... - ..... ------- ............ Neburon EPA-2
Kop-Thion ---- ............... - ............... Ma lath ion EPA-3
krysid (Russia)--- ..... ---------- ..... ------ Antu EPA-1
Kwik-ki 1 ------------------------------------ Strychnine EPA-1*
Kypfar in ------------------------------------ Warfar in EPA-^4
Kypfos- ....... ----- ..... --- ....... -- ....... -Ma lath ion EPA-3
L 3*314 --------- ........................ ---- Diphenamid EPA-1 & EPA-2
Lab i 1 i te ------------------------ • ------------ Thiophanate-methyl EPA-1
Lamprecid (Trade Name) EPA-1 ---------------- LIV
Leivasom ------------------------------------ Trichlorfon EPA-3 6 EPA-4
Liphadione ---------------------------------- Chlorophac inone EPA-2 £ EPA-3
LM 91--- .......... - ....... - ................. Chlorophacinone EPA-2 & EPA-3
M Ik (USSR) .................. - .............. Disulfoton EPA-3
Maintain A ---------------------------------- Chlorof lurecol-methyl ester EPA-1
Maintain CF 125-- ---------- ..... --- ...... --- Chi oroflurecol -methyl ester EPA-1
Maintain S ...... --- ...... - ...... - ....... ---- Chlorof lurecol -methyl ester EPA-1
Malamar ------------------------------------- Ma lath ion EPA-3
Malaphele- ........ - ....... ------- ..... - ..... Ma lath ion EPA-3
Mai asp ray ----------------------------------- Ma lath ion EPA-3
Malathion EPA-3 ---- ......................... HPLC ( I S)
Malathion ULV Concentrate ------------------- Malathion EPA-3
Malatol ------ ..... -------------------------- Malathion EPA-3
maldison (Australia) ------------------------ Malathion EPA-3
Maimed' -------------------------------------- Malathion EPA-3
Maltox ....... - ............................. -Malathion EPA-3
mercaptothion (So. Africa) ------------------ Malathion EPA-3
mercaptothion (Argentina) ------------------- Malathion EPA-3
Merpan ------ ...... ------ ...... -------------- Captan EPA-3 & EPA-4
Metacide ------------------------------------ Methyl pa rath ion EPA-6
-------
10
Methazole EPA-1 -j_R
Methidathion EPA-2- - - -GC-FID-IS
Methoprene EPA-1- - -GC-FID-IS
S-[(5-methoxy-2-oxo-1,3,4-th iadiazol-
3(2H)-yl)methyl]0,0-d!methyl
phosphorodithioate --Methidathion EPA-2
Methyl-2-chloro-9-hydroxyfluorene-9-
carboxylate Chloroflurecol-methyl ester EPA-1
N-(2-methyl-4-chlorophenyl)-N',N'-
dimethyl formamidine Chlordimeform EPA-1
2,2-methylenebis (3,^,6-
tr ichlorophenol) Hexachlorophene EPA-1
Methyl nonyl ketone EPA-1 GC-FID-IS
Methyl parathion EPA-6-- -HPLC
3-( 1-methyl -2-pyrrol i dyl )pyridine- Nicotine EPA-1
metolachlor (with atrazine) Mixed pesticides EPA-2
metrifonate Trichlorfon EPA-3 & EPA-4
Metron Methyl parathion EPA-6
Mezene Zi ram EPA-1
MGK Dog and Cat Repellent- Methyl nonyl ketone EPA-1
Microzul Chlorophacinone EPA-2 & EPA-3
Mi 1 do thane Th iophanate-methyl EPA-1
Mi locep metolachlor
Mixed pesticides EPA-2
(Atrazine & Metolachlor) GC-FID- IS
Mixed pesticides EPA-3
(Disulfoton & Fensulfothion) GC-FID- IS
MLT - Ma lath ion EPA-3
Mouse-tox Strychnine EPA-4
Multimet Bend i oca rb EPA-3
Mult iprop Chlorof lurecol-methyl ester EPA-1
N 2790 Fonofos EPA-1
NAA Naphthaleneacetic acid EPA-1
NAA 800 Naphthaleneacetic acid EPA-1
Nabac Hexachlorophene EPA-1
-------
11
Mac -- Carbaryl EPA-1
Nafusaku Naphthaleneacetic acid EPA-1
1-naphthal eneacet ic acid Naphthal eneacet ic acid EPA-1
N-1-naphthyl phthalamic acid Naptalam EPA-1
Naphthal eneacet ic acid EPA-1 HPLC
1-naphthly-N-methylcarbamate Carbaryl EPA-3
alpha-naphthy1 thiourea Antu EPA-1
Naptalam EPA-1--- IW
NC 6897 -- - Bendiocarb EPA-3
Neburex Neburon EPA-2
Neburon EPA-2--- --- UV
Neguvon Trichlorfon EPA-3 & EPA-4
neko Rotenone EPA-2
NF 35 -- - —-Thiophanate EPA-1
Nicocyan Pendimethal in EPA-1
Nicotine EPA-1 HPLC (I S)
nicoul ine Rotenone EPA-2
J»-nitro-3-(trif luoromethyl )pheny 1 Lamprecid (Trade Name) EPA-1
Nitrox 80 Methyl parathion EPA-6
NPA- — --- - Naptalam EPA-1
On track 8E metolachlor
Orthocide Captan EPA-3 & EPA-A
oxydiazol Methazole EPA-1
Para-Col Paraquat EPA-1
Paraquat EPA-1 - HPLC (IS)
Parataf Methyl parathion EPA-6
Paratox Methyl parathion EPA-6
Partron M Methyl parathion EPA-6
Pathclear Paraquat EPA-1
Pay-Off - Pendimethal in EPA-1
P.C.Q. - Diphacinone EPA-3
penchoral Pentachlorophenol EPA-1 & EPA-2
Pendimethal in EPA-1 GC-FID-IS
-------
12
Penncap-M Methyl pa rath ion EPA-6
penoxal i n Pendimethal in EPA-1
penoxyn Pendimethal in EPA-1
Pentachlorophenol EPA-1 GC-FIP- IS
Pentachlorophenol EPA-2 HPLC ( IS)
Pentacon Pentachlorophenol EPA-1 & EPA-2
Penwar Pentachlorophenol EPA-1 6 EPA-2
Phyomone Naphthaleneacetic acid EPA-1
Pi 1 larquat Paraquat EPA-1
Pi larxone Paraquat EPA-1
Planof ix Naphtha! eneacet ic acid EPA-1
Plucker Naphtha leneacetic ac id EPA-1
polychlorinated dibenzo-p-dioxin Dioxins EPA-1
polychlor inated dioxins • Dioxins EPA-1
Pomarsol I Forte Ziram EPA-1
Prentox Rotenone EPA-2
Priltox Pentachlorophenol EPA-1 £ EPA-2
Primacol Naphtha leneacetic acid EPA-1
Pr i mag ram metolachlor
Pr i mat ex • metolachlor
Primatol A Atrazine EPA-3 6 EPA-4
Probe Methazole EPA-1
Prodaram--- Ziram EPA-1
Promar • Diphacinone EPA-1
propanoic acid Propionic acid EPA-1
Propionic acid EPA-1- • GC-FID
Propionic Acid Grain Preserver Propionic acid EPA-1
Prowl Pendimethal in EPA-1
Proxol - - Trichlorfon EPA-3 & EPA-**
6Q8 - Naptalam EPA-1
Rabon Tetrachlorvinphos EPA-1
Rabone Tetrachlorvinphos EPA-1
Ramik Diphac inone EPA-3
Ramucide Chlorophacinone EPA-2 & EPA-3
-------
13
Ratomet - --Chlorophacinone EPA-2 & EPA-3
Ratox Warfarin EPA-4
Raviac Chlorophacinone EPA-2 & EPA-3
Ravyon Carbaryl EPA-3
RAX- - -Warfarin EPA-4
RDL Tetrachlorovinphos EPA-1
Reg]ox Diquat EPA-1
region (USSR) - Diquat EPA-1
Reg lone Diquat EPA-1
Ro-Dec Strychnine EPA-4
Rodent Cake Diphacinone EPA-3
Rodex Warfar in EPA-4
Rodex Blox Warfarin EPA-A
Rootone Naphthal eneacet ic acid EPA-1
Rotenone EPA-2 HPLC
Rozol Chlorophacinone EPA-2 & EPA-3
S 767 fensulfothion
Santobrite Pentachlorophenol EPA-1 & EPA-2
Santophen • Pentachlorophenol EPA-1 & EPA-2
SD 3562 - - — Dicrotophos EPA-1
SD 8H7 Tetrachlorvinphos EPA-1
Sentry Grain Preservei Propionic acid EPA-1
Sept en e Carbaryl EPA-3
Sevin Carbaryl EPA-3
sevin (USA, USSR) Carbaryl EPA-3
Shell Atrazine Herbicide Atrazine EPA-3 £ EPA-4
Sigma Th iophanate-methy 1 EPA-1
Sinbar Terbaci 1 EPA-1
Sinituho Pentachlorophenol EPA-1 6 EPA-2
SN 36268 - - Chlordimeform EPA-1
Sol vi rex Disulfoton EPA-2
Spa none Chlordimeform EPA-1
-------
Spike Tebuthiuron EPA-1
St ik Naphthaleneacetic acid EPA-1
St irofos Tetrachlorvf nphos EPA-1
Strychnine EPA-**- - HPLC (IS)
Strychnos nux-vomica • Strychnine EPA-4
Sumi tox Ma lath ion EPA-3
Supracide - --Methi dathion EPA-2
Sweep Paraquat EPA-1
TCNB -- -Tecnazene EPA-1
Tebuthiuron EPA-1- UV
Tecnazene EPA-1- -GC-FID-IS
Tekkam • Naphthaleneacetic acid EPA-1
Tekwa i sa Methyl pa rath ion EPA-6
Terbacil EPA-1- UV
Tercyl - Carbaryl EPA-3
Terracur P Fensulfothion
Terraklene Paraquat EPA-1
Tersan SP Chloroneb EPA-1
1,2,4,5-tetrach1oro-3-nitrobenzene- Tecnazene EPA-1
2,3,5,6-tetrachloronitrobenzene Tecnazene EPA-1
Tel:rachlorv?nphos EPA-1 GC-FID-IS
JPM -Lamprecid (Trade Name) EPA-1
Thiophanate EPA-1- - -UV
thiophanate-ethyl Thiophanate EPA-1
Th'rophanate-methyl EPA-1 UV
TipOff Naphthaleneacetic acid EPA-1
Topitox Chlorophacinone EPA-2 & EPA-3
Tops in Thiophanate EPA-1
Tops in E Thiophanate EPA-1
Tops in M Thiophanate-methyl EPA-1
TotaCol Paraquat EPA-1
Toxer Total Paraquat EPA-1
Tox-Hid Warfar in EPA-4
Transplantone Naphthaleneacetic acid EPA-1
-------
15
Tre-Hold Naphtha!eneacetic acid EPA-1
Tricarnam Carbaryl EPA-3
Tricarbamix Z Ziram EPA-1
Trichlorfon EPA-3-- -GC-FID-IS
Trichlorfon EPA-4- --HPLC (IS)
trichlorphon Trichlorfon EPA-3 & EPA-4
ci s-N-tr ichloromethylthio-4-
cyclohexane-1,2-dicarboximi de Captan EPA-3 & EPA-4
alpha, alpha, alpha-trifluoro-4-
nitro-meta-cresol Lamprecid (Trade Name) EPA-1
3-trif 1 uoro-4-ni trophenol Lamprecid (Trade Name) EPA-1
Trinex Trichlorfon EPA-3 & EPA-4
Tr iscabol Ziram EPA-1
tub at ox in Rotenone EPA-2
Tugon - - Trichlorfon EPA-3 & EPA-4
UC 7744 - - Carbaryl EPA-3
Ultracide - Methidathion EPA-2
2-urtdecanone --Methyl nonyl ketone EPA-1
USB 3584 Dinitramine EPA-1
Vancide MZ-96 - Ziram EPA-1
VCS 438— - Methazole EPA-1
Vectal SC - Atrazine EPA-3 & EPA-4
Vegfru Ma lath ion EPA-3
Vertac Methyl Parathion Technisch 80% Methyl parathion EPA-6
Voncaptan Captan EPA-3 & EPA-4
Warfarin EPA-4- - HPLC
Warfarin Plus Warfarin EPA-4
Warfarin Q Warfarin EPA-4
Weedol Paraquat EPA-1
Weedone Pentachlorophenol EPA-1 £ EPA-2
Weedtrine D Diquat EPA-1
Wofatox Methyl parathion EPA-6
-------
16
Z-C Spray Ziram EPA-1
Zerlate - —Ziram EPA-1
zinc dimethyldi thiocarbamate Ziram EPA-1
Zincmate Zi ram EPA-1
Ziram EPA-1 ~UV
Zi ramvis Zi ram EPA-1
Zirasan 90 Ziram EPA-1
Zirberk - Ziram EPA-1
Zirex 90 - Ziram EPA-1
Ziride Ziram EPA-1
Zithol Ziram EPA-1
Zitox-- Ziram EPA-1
zoocoumarin (Netherlands & USSR) Warfarin EPA-1*
ZR 515 - Methoprene EPA-1
-------
Analytical Methods - Introduction
Many of the methods in this manual have been developed and are
used by chemists in state and federal regulatory laboratories. Some
are "old-time methods" that have been used over the years, and some
are "new methods" recently developed to utilize new instrumentation
or to analyze for new compounds in new formulations.
These methods have been written in a relatively standard format
for several reasons:
(1) to allow the neophyte pesticide chemist to more easily
understand and perform the various analyses
(2) to provide a standardized form so that the validity and
application of the method can be more easily evaluated
by the experienced pesticide chemist
(3) to allow changes in one or more sections without entirely
rewriting the entire method
The editorial committee welcomes the submission of new methods
and especially the correction of errors, criticism, suggestions with
supporting data, new ideas, and general comments on the published
methods.
-------
Note: Throughout these methods, the term "teflon" has been used to
denote Teflon, the registered trademark of E. I. du Pont de
Nemours & Co. for chemically resistant fluorocarbon resin.
t
t
-------
3rd Update - August 1982
Antu EPA - 1
Determination of Antu by Ultraviolet Spectroscopy
Antu is the accepted (BSI, ISO) common name for alpha-naphthylthiourea, a
registered rodenticide having the chemical structure:
NH C-—NH,
Molecular formula: ^11^10^2^
Molecular weight: 202.3
Physical state-color-odor: pure - white crystals
technical - blue-gray powder
Melting point: 198°C
Solubility: 0.06 gram per 100 ml in water at room temperature;
8.6 grams per 100 ml in triethylene glycol; 2.^3 grams
per 100 ml in acetone
Stability: stable on exposure to air or sun
Other names: krysid (Russia)
Reagents;
1. Antu standard of known purity
2. Methanol, pesticide or spectro grade
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
-------
3rd Update - August 1982 Antu EPA - 1
Procedure;
Preparation of standard;
Weigh 80 mg antu standard into a TOO ml volumetric flask, dissolve in and make
to volume with methanol; mix thoroughly. Pipette 5 ml into a second 100 ml
volumetric flask and make to volume with methanol. Mix thoroughly and pipette
5 ml into a third 100 ml volumetric flask and make to volume with methanol;
mix thoroughly, (final cone 2 ug/ml)
Preparation of sample;
Weigh a portion of sample equivalent to 80 mg antu into a 250 ml glass-
stoppered flask or screw-cap bottle. Add 100 ml methanol by pipette, stopper
tightly and shake on a mechanical shaker for thirty minutes. Allow to settle,
centrifuge or filter if necessary, taking precautions to avoid evaporation of
the solvent. Pipette 5 ml into a 100 ml volumetric flask, make to volume with
methanol, and mix thoroughly. Pipette 5 ml into another 100 ml volumetric
flask, make to volume with methanol, and mix thoroughly.
(final cone 5 ug antu/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative settings for the
particular instrument being used, balance the pen at 0 and 100% transmission
with methanol in each cell. Scan both standard and sample solutions from 3&0
to 200 nm with methanol in the reference cell. Measure the absorbance of
standard and sample solutions using the maxima at 220 nm and a baseline at
360 nm. If interference by the inert ingredients is suspected at 220 nm, make
measurements at 281 nm. The concentration should be increased to 10 ug/ml for
measurement at 281 nm.
Calculation;
From the above absorbances and using the standard and sample concentrations,
calculate the percent antu as follows:
(abs. sample) (cone. std. in ug/ml) (% purity)
(abs. std.) (cone, sample in ug/ml)
Absorbance is linear dt both wavelengths when plotted against concentration.
Method submitted by EPA (former) Product Analysis Laboratory, Region II, New
York, NY
January 1977
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982 Atrazine EPA-3
Determination of Atrazine by High Performance Liquid Chromatography
Atrazine is the accepted (ANSI, BSI, ISO, WSSA) common name for 2-chloro-
4-ethylamino-6-isopropylamino-1,3,5-triazine, a registered herbicide having
the chemical structure:
Cl
I
C
HI H
I I II I
CH3 CH2 N C^ XC N CH"
\
N
Molecular formula: CgH^rClNr
Molecular weight: 215.7
Physical state-color-odor: colorless crystalline solid
Melting point: 173 to 175°C
Solubility: 33 ppm in water at 25°C; 1.2% in ethyl ether, 1.8% in methanol,
2.8% in ethyl acetate, 5.2% in chloroform, 18.3% in dimethyl
sulfoxi de
Stability: stable in neutral and slightly acidic or basic media; hydrolyzes
in acid and alkaline conditions of higher temperatures to the
herbicidally inactive hydroxy derivative; non-flammable; non-
corrosive under normal use conditions; very stable shelf life with
only slight sensitivity to natural light and extreme temperature;
compatible with most other pesticides
Other names: AAtrex; Atranex; Atratol; Crisatrina; Crisazine; G 30027;
Gesaprim; Griffex; Primatol A; Shell Atrazine Herbicide;
Vectal SC; 6-chIoro-N-ethyl-N'-(l-methylethyl)-1,3,5-triazine-
2,^-diamine
-------
3rd Update - August 1982 Atrazine EPA-3
Reagents;
1. Atrazine standard of known purity
2. Methanol, HPLC or pesticide grade
3- Acetic acid, ACS
Equipment:
1. High Performance Liquid Chromatograph with UV detector at 254 nm. If a
variable wavelength detector is available, other wavelengths may be used
to increase sensitivity or to eliminate interference.
2. Column: Radial Pak C18 (or equivalent column and parameter adjustments)
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 851 methanol + 14% water + 1% acetic acid
Column temperature: ambient
F1ow ra te: 7 m1/m i n
Wavelength: 254 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg atrazine standard into a 25 ml volumetric flask, dissolve in and
make to volume with methanol, stopper tightly, and place in an ultrasonic bath
until completely dissolved. Filter a portion through a 0.45 micron filter.
(cone 4 mg/ml)
Preparation of sample;
Weigh a portion of sample equivalent to 100 mg atrazine into a 25 ml
volumetric flask, make to volume with methanol and place in an ultrasonic bath
as above. Filter a portion through a 0.45 micron filter, (cone 4 mg/ml)
-------
3rd Update - August 1982 Atrazine EPA-3
HPLC Determination;
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample solutions.
Calculat ion:
Measure the peak height or peak area for each peak and calculate the average
for both standard and sample. Using these averages, calculate the percent
atrazine as follows:
(peak height or area sample)(weight standard injected)(% purity standard)
(peak height or area standard)(weight sample injected)
This method had its origin at the Beltsville Chemistry Lab but is a result of
several modifications from the HPLC courses sponsored by EPA - Beltsville and
NEIC, Denver, Colorado
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982 Atrazine EPA-A
Determination of Atrazine by High Performance Liquid Chromatography
Atrazine is the accepted (ANSI, BSI, ISO, WSSA) common name for 2-chloro-
4-ethylamino-6-isopropylamimo-1,3,5-triazine, a registered herbicide having
the chemical structure:
Cl
CH~' CH^ —
*
H
1
— N (
H CH3
_C N CH
^CH,
Molecular formula: CgH^ClN,-
Molecular weight: 215.7
Physical state-color-odor: colorless crystalline solid
Melting point: 173 to 175°C
Solubility: 33 ppm in water at 25°C; 1.2% in ethyl ether; 1.8% in methanol;
2.8% in ethyl acetate; 5.2% in chloroform; 18.3% in dimethyl
sulfoxide
Stability: stable in neutral and slightly acidic or basic media; hydrolyzes
in acid and alkaline conditions of higher temperatures to the
herbicidally inactive hydroxy derivative; non-flammable; non-
corrosive under normal use conditions; very stable shelf life
with only slight sensitivity to natural light and extreme
temperature; compatible with most other pesticides
Other names: AAtrex; Atranex; Atratol; Crisatrina; Crisazine; G 30027;
Gesaprim; Griffex; Primatol A; Shell Atrazine Herbicide;
Vectal SC; 6-chloro-N-ethyl-N'-(1-methylethyl)-1,3,5-triazine-
2,^-diamine
-------
3rd Update - August 1982 Atrazine EPA-4
Reagents:
1, Atrazine standard of known purity
2. Diethyl phthalate (internal standard) of known purity
3. Acetonitrile, HPLC grade
4. Methanol, HPLC grade
5. Internal standard solution - weigh 200 rug diethyl phthalate into a 100 ml
volumetric flask, dissolve in and make to volume with methanol; mix well.
(cone 2 mg/ml)
Eg u i pmen t:
1. High Performance Liquid Chromatograph with UV detector at 25^ nm. If a
variable wavelength detector is available, other wavelengths may be used
to increase sensitivity or to eliminate interference.
2. Column: uBondapak C18 (30 cm x 3-9 rnm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
A. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: k2% acetonitrile + 58% water
Column temperature: 33°C
F1ow ra te: 2.5 m1/m i n
Wavelength: 254nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg atrazine standard into a 125 ml screw-cap flask, add 100 ml
internal standard solution by pipette, close tightly, and shake to dissolve.
Filter a portion through a 0.45 micron filter, (cone 1 mg atrazine and 2 mg
diethyl phthalate per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 100 mg atrazine into a 125 ml screw-
cap flask and add 100 ml internal standard solution by pipette. Close
tightly, shake a few minutes by hand, place in an ultrasonic bath for about
2 minutes, and shake on a mechanical shaker for one hour. Filter a portion
through a 0.45 micron filter, (cone as above)
-------
3rd Update - August 1982 Atrazine EPA-*t
HPLC Determination;
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample solutions.
Calculat ion;
Measure the peak heights or areas of the atrazine and the diethyl phthalate
for both the standard and sample solutions and calculate the following ratios:
peak height or area atrazine
Ratio of standard = -
Ratio of sample
peak height or area diethyl phthalate
peak height or area atrazine
peak height or area diethyl phthalate
Average the standard and sample ratios, and calculate the percent atrazine as
follows:
(ratio of sample) (weight standard) (% purity of standard)
(ratio of standard) (weight sample)
Method submitted by EPA - NEIC, Denver, Colorado (Chuck Rzeszutko) December
1979
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982
Bendiocarb EPA-3
Determination of Bendiocard by High Performance Liquid Chromatography
Bendiocarb is the accepted (ANSI, BSI, ISO) common name for 2,2-dimethyl •
1,3-benzodioxol-k-y] N-methylcarbamate, a registered insecticide having the
chemical structure:
0 C N CH.
0 H
Molecular formula:
Molecular weight: 223.23
Physical state-color-odor: white crystalline solid
Melting point: 129 to 130°C
Solubility: at 25°C: 0.004% in water, 0.03% in kerosene, 1.0% in o-xylene,
k% in ethanol and benzene, and 20% in acetone, dichloromethane,
dioxin, and chloroform
Stability: the hydrolysis (to the phenol) half-life in solution in 0.01M
aqueous sodium phosphate buffer at pH 7 and 25°C is 20 days
Other names: Ficam; Garvox; Multimet; NC 6897; 2,3-isopropylidenedioxyphenyl
methylcarbamate
Reagents:
1. Bendiocarb standard of known purity
2. Methyl benzoate (internal standard) of known purity
3. Methanol, HPLC grade
k. Water, HPLC grade
-------
3rd Update - August 1982 Bendiocarb EPA-3
5. Internal standard solution - weigh 180 mg methyl benzoate into a 250 ml
volumetric flask, dissolve in and make to volume with methanol; mix
thoroughly, (cone. 0.72 mg/ml)
Equipment;
1. High Performance Liquid Chromatograph with a variable wavelength UV
detector at 280 nm. If a variable wavelength detector is not available
operating parameters and concentrations may have to be changed to obtain
the necessary separation and sensitivity.
2. Column: uBondapak C18 (30 cm x 3.9 mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loops 10 ul
k. Mechanical shaker and/or ultrasonic bath
5. 0.^5 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 50£ methanol + 50 % water
Column temperature: ambient
Flow rate: 2 ml/min
Wavelength: 280 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 110 mg bendiocarb standard into a 100 ml volumetric flask, dissolve in
and make to volume with methanol; mix thoroughly. Pipette a 10 ml aliquot
into a 50 ml volumetric flask, add 20 ml internal standard solution by
pipette, and make to volume with methanol. Mix thoroughly and filter a
portion through a 0.^5 micron filter, (cone .0.22 mg bendiocarb and 0.288 mg
methyl benzoate per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 110 mg bendiocarb into a 100 ml
volumetric flask, dissolve in and make to volume with methanol; mix
thoroughly. Pipette a 10 ml aliquot into a 50 ml volumetric flask, add 20 ml
internal standard solution by pipette, and make to volume with methanol. Mix
thoroughly and filter a portion through a 0.^5 micron filter, (cone as above)
-------
3rd Update - August 1982 Bendiocarb EPA-3
HPLC Determination;
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Proceed with the determination making alternately three injections each
of standard and sample solutions. Elution order is bendiocarb then methyl
benzoate.
Calculat ion;
Measure the peak heights or areas of the bendiocarb and the methyl benzoate
for both the standard and sample solutions and calculate the following ratios:
peak height or area bendiocarb
Ratio of standard =
peak height or area methyl benzoate
peak height or area bendiocarb
Ratio of sample = —•—— ——
peak height or area methyl benzoate
Average the standard and sample ratios, and calculate the percent bendiocarb
as follows:
(ratio of sample) (weight standard) (% purity of standard)
(ratio of standard)(weight sample)
Method submitted by EPA - NEIC, Denver, Colorado (M. Sher Ali)
November 1980
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
-------
3rd Update - August 1982 Captan EPA-3
Determination of Captan by Gas Chromatography (FID-IS)
Captan is the common name for cis-N-trichloromethylthio-^-cyclo-hexene-
1,2-dicarboximide, a registered fungicide having the chemical structure:
0
Cl
Molecular formula: CgHg
Molecular weight: 300.6
Physical state-color-odor: (pure) - white crystalline solid
(technical) - yellow amorphous solid with a
pungent odor
Melting point: (pure) - 178°C (decomposes)
(technical) - 160 to 170°C (93 to 95% purity)
Solubility: less than 0.05 ppm in water at room temperature; insoluble in
petroleum oils; at 25°C the solubility w/w is 1% in xylene, 5
chloroform, 3% in acetone, and 1% in isopropanol
Stability: stable under alkaline conditions; decomposes at its melting point;
non-corrosive but decomposition products are corrosive
Other names: captane (France); Merpan; Orthocide; Voncaptan
Reagents;
1. Captan standard of known purity
2. Dibutyl phthalate (internal standard), analytical grade
3. Acetone, pesticide grade
-------
3rd Update - August 1982 Captan EPA-3
Internal standard solution - weigh 100 mg dibutyl phthalate into a 100 ml
volumetric flask, dissolve in and make to volume with acetone; mix well.
(cone 1 mg/ml)
Equi pmentt
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 1/V glass packed with 3% OV-1 on 100 to 120 mesh Supelcoport
3. Precision liquid syringe
4. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Ultrasonic bath
7. Usual laboratory glassware
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Operating conditions for FID;
Column temperature: 180°C
Injection port temperature: 250°C
Detector temperature: 250°C
Carrier gas: nitrogen - 30 ml/min (adjusted as necessary)
Hydrogen flow: 30 ml/min (adjusted as necessary)
Air flow: 600-800 ml/min (adjusted as necessary)
Procedure;
Preparation of standard;
Weigh 75 mg captan standard into a small (30 to 40 ml) glass vial with a
polyseal-1ined cap, add 25 ml internal standard solution by pipette, and close
tightly. Shake for several minutes and place in a sonic bath for about 2
minutes and shake a few minutes more. Allow to settle, centrifuge to settle-
out the particulates. (cone 3 mg captan and 1 mg dibutyl phthalate per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 75 mg captan into a small vial as
above and follow the same procedure, (cone - as above)
GC Determination;
Inject 2 ul of standard and, if necessary, adjust the instrument parameters
and the volume injected to give a complete separation within a reasonable time
and to obtain peak heights of 1/2 to 3/4 full scale. Proceed with the
determination, making at least three injections each of standard and sample
-------
3rd Update - August 1982 Captan EPA-3
solutions. The elution order is dibutyl phthalate then captan.
Calculation;
Measure the peak areas of the captan and dibutyl phthalate for both the
standard and sample solutions and calculate the following ratios:
peak area captan
Ratio of standard =
Ratio of sample
peak area dibutyl phthalate
peak area captan
peak area dibutyl phthalate
Average the standard and sample ratios, and calculate the percent captan as
follows:
(ratio of sample) (weight of standard) (% purity of standard)
(ratio of standard)(weight of sample)
Method submitted by NEIC, Denver, Colorado (Chuck Rzeszutko), June 1979
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982
Captan EPA-*t
Determination of Captan by High Performance Liquid Chroma tography
Captan is the common name for cis-N-trichloromethylthio-^-cyclo-hexene-
1 ,2-dicarboximide, a registered fungicide having the chemical structure:
Cl
I
C Cl
Cl
Molecular formula:
Molecular weight: 300.6
Physical state-color-odor: pure - white crystalline solid
technical - yellow amorphos solid with a
pungent odor
Melting point: pure - 178°C (decomposes)
technical - 160 to 170°C (93 to 95* purity)
Solubility: less than 0.05 ppm in water at room temperature; insoluble in
petroleum oils; at 25°C the solubility w/w is 1% in xylene, 5%
in chloroform, 3& in acetone, and 1% in isopropanol
Stability: stable under alkaline conditions; decomposes at its melting point;
non-corrosive but decomposition products are corrosive
Other names: captane (France); Merpan; Orthocide; Voncaptan
Reagents;
1. Captan standard of known purity
2. Dibutyl phthalate (internal standard) of known purity
-------
3rd Update - August 1982 Captan EPA-^
3. Methylene chloride, HPLC grade, dried with anhydrous sodium sulfate
*». Water, HPLC grade
5. Internal standard solution - weigh 150 mg dibutyl phthalate into a 500 ml
volumetric flask, dissolve in and make to volume with dried methylene
chloride; mix well. (0.3 mg/ml)
Equipment;
1. High Performance Liquid Chromatograph with UV detector at 25^ nm. If a
variable wavelength detector is available, other wavelengths may be used
to increase sensitivity or to eliminate interference.
2. Si-100 10 urn (Spectra-Physics) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
k. Mechanical shaker and/or ultrasonic bath
5. 0.kS micron f i iter
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 100% methylene chloride (dried with anhydrous sodium sulfate)
Column temperature: ambient
Flow rate; 1.5 ml/min
Wavelength: 25^ nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure;
Preparation of standard;
Weigh 50 mg captan standard into a 50 ml volumetric flask, dissolve in and
make to volume with internal standard solution; mix thoroughly and filter a
portion through a O.A5 micron filter, (cone 1 mg captan and 0.3 mg dibutyl
phthalate per ml)
Preparation of sample;
Weigh a portion of liquid sample Into a 50 ml volumetric flask, make to volume
with internal standard solution, and mix well. For solid or dry samples,
weigh a portion equivalent to 50 mg captan into a 125 ml screw-cap flask and
add 50 ml internal standard solution by pipette, and mix well. Filter a
portion through a 0.^5 micron filter, (cone as above)
-------
3rd Update - August 1982 Captan EPA-'t
HPLC Determination:
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Proceed with the determination making alternately three injections each
of standard and sample solutions.
Calculat ion:
Measure the peak heights or areas of the captan and the dibutyl phthalate for
both the standard and sample solutions and calculate the following ratios:
peak height or area captan
Ratio of standard = —
Ratio of sample =
peak height or dibutyl phthalate
peak height or area captan
peak height or area dibutyl phthalate
Average the standard and sample ratios, and calculate the percent captan as
follows:
(ratio of sample) (weight standard) (% purity standard)
(ratio of standard)(weight sample)
Method submitted by Mark W. Law, EPA Beltsville Chemistry Lab, Beltsville, MD
April 1980
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982
Carbaryl EPA-3
Determination of Carbaryl by High Performance Liquid Chromatography
Carbaryl is the accepted (BSI, ISO) common name for 1-naphthyl-N-
methylcarbamate, a registered insecticide having the chemical structure:
0-
C N
II I
0 H
Molecular formula:
Molecular weight: 201.2
Physical state-color-odor: white crystalline solid
Melting point:
Solubility: AO ppm in water at 30°C; soluble in most polar organic solvents
such as acetone and mixed cresols
Stability: stable to light, heat, and hydrolysis under normal storage
conditions; non-corrosive to metals, packing materials, or
application equipment; compatible with most pesticides except
those strongly alkaline which hydrolyze it to 1-naphthol
Other names: Carbamine; Cekubaryl; Denapon; Devicarb; Dicarbam; Hexavin;
Karbaspray; Nac; Ravyon; Septene; Sevin; sevin (USA, USSR);
Tercyl; Tricarnam; UC 77^
Reagents;
1. Carbaryl standard of known purity
2. Diethyl phthalate (internal standard) of known purity
3. Acetonitrile, HPLC grade
k. Methanol , HPLC grade
-------
3rd Update - August 1982 Carbaryl EPA-3
5. Internal standard solution - weigh 2 grams diethyl phthalate into a 250 ml
volumetric flask, dissolve in and make to volume with methanol; mix
thoroughly, (cone 8 mg/ml)
Eg u i pme n t;
1. High Performance Liquid Chromatograph with a variable wavelength UV
detector at 280 nm. If a variable wavelength detector is not available,
operating parameters and concentrations may have to be changed to obtain
the necessary separation and sensitivity.
2. Column: uBondapak C18 (30 cm x 3-9 mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions^:
Mobile phase: 50% acetonitrile + 50% water
Column temperature: 30°C
Flow rate: 2 ml/min
Wavelength: 280 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedu re:
Preparation of standard;
Weigh 100 mg carbaryl standard into a 125 ml screw-cap flask, add 100 ml
internal standard by pipette, close tightly, and shake to dissolve.
Filter a portion through a 0.^5 micron filter, (cone 1 mg carbaryl and 8 mg
diethyl phthalate per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 100 mg carbaryl into a 125 ml screw-
cap flask, add 100 ml internal standard by pipette, close tightly, and shake
to dissolve the carbaryl . A few minutes in an ultrasonic bath may help to
disperse and dissolve the sample. Filter through a 0.^5 micron filter.
(cone as above)
-------
3rd Update - August 1982 Carbaryl EPA-3
HPLC Determination;
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample solutions. Elution order is carbaryl then diethyl
phthalate.
Calculation;
Measure the peak heights or areas of the carbaryl and the diethyl phthalate
for both standard and sample solutions and calculate the following ratios:
peak height or area carbaryl
Ratio of standard = •
Ratio of sample
peak height or area diethyl phthalate
peak height or area carbaryl
peak height or area diethyl phthalate
Average the standard and sample ratios, and calculate the percent carbaryl as
follows:
(ratio of sample) (weight standard) (% purity of standard)
(ratio of standard) (weight sample)
Method submitted by EPA - NEIC, Denver, Colorado (Chuck Rzeszutko)
August 1979
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982
Chlorof1urecol-methyl ester EPA-1
Determination of Chlorof1urecol-methyl ester in Liquid
Formulations by Ultraviolet Spectroscopy
Chlorof1urecol-methyl ester is the common name (approved by BSI and used
in the United States and Great Britain) for methyl-2-chloro-9-hydroxyfluorene-
9-carboxylate, a registered plant growth regulator having the chemical
structure:
[Technical Maintain CF 125 has 65 - 70% of the above compound plus:
18 - 25% methyl-9-hydroxyfluorene-9-carboxylate
10 - 12% methyl-2,7-dichloro-9-hydroxyfluorene-9-carboxylate
Since most formulations contain all three isomers, a technical Maintain CF 125
standard must be used for analysis.]
Molecular formula:
Molecular weight: 274.7
Physical state-color-odor: odorless, white crystals when pure
Melting point: 152°C
Solubility: grams per 100 ml solvent at 20°C: acetone - 26, benzene - 8,
carbon tetrachloride - 2.4, cyclohexane - 0.24, ethanol - 8,
isopropanol - 2.4, methanol - 15, petroleum ether
(bp 50-70°C) - 0.16, water - 0.00218
Stability: stable at room temperature; compatible with other growth
regulators and with NH 30
-------
3rd Update - August 1982 Chloroflurecol-methyl ester EPA-1
Other names: chloroflurenol-methyl ester (ISO and France); CF 125;
curbiset; IT 3^56; Maintain CF 125; Maintain A; Maintain S;
Multiprop
Reagents;
1. Maintain CF 125 standard of known assay
2. Methanol, pesticide or spectro grade
Equi pment;
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
k. Usual laboratory glassware
Procedures;
Preparation of standard;
Weigh 75 mg Maintain CF 125 standard into a 100 ml volumetric flask, dissolve
in and make to volume with methanol, and mix thoroughly. Pipette 10 ml into
a second 100 ml volumetric flask, make to volume with methanol, and mix
thoroughly. Pipette 10 ml into a third 100 ml volumetric flask, again make to
volume with methanol and mix thoroughly, (final cone 7.5 ug/ml)
Preparation of sample;
Weigh a portion of sample equivalent to 75 mg Maintain CF 125 into a 100 ml
volumetric flask, make to volume with methanol, and mix thoroughly. Make a
second and third dilution as above to give a final concentration of 7-5 ug/ml.
UV Determinat ion;
With the UV spectrophotometer at the optimum quantitative settings for the
particular instrument being used, balance the pen at 0 and 100% transmission
at 275 rim with methanol in each cell. Scan both standard and sample solutions
from 350 to 230 nm with methanol in the reference cell. Measure the
absorbance of standard and sample solutions at 275 nm using a reference point
at 350 nm.
-------
3rd Update - August 1982 Chloroflurecol-methyl ester EPA-1
Calculat ions;
From the above absorbances and using the standard and sample concentrations,
calculate the percent Maintain CF 125 as follows:
(abs. sample) (cone. std. in ug/ml) (% purity standard)
(abs. std.) (cone, sample in ug/ml)
There is a straight line relationship between absorbance and concentration for
up to 12.5 ug/ml.
Method submitted by EPA (former) Product Analysis Laboratory, Region II, New
York, NY
November 1977
Any criticisms, suggestions, or data concerning this method will be
appreciated.
-------
3rd Update - August 1982 Chlordimeform EPA-1
Determination of Chlordimeform by Gas Chromatography (FID-IS)
Chlordimeform is the accepted (ANSI, BSI, ISO) common name for
N'-(4-chloro-o-tolyl)-N,N-dimethyl formamidine, a registered acaricide,
insecticide, and ovicide having the chemical structure:
H
CH3
CH3
CH3
Molecular formula: base - C10H1-C1N2 HC1 salt - CioHUC12N2
Molecular weight: base - 196.7 HC1 salt - 233.1
Physical state-color-odor: colorless crystals with a faint
amine-1 ike odor (both base and HC1
salt); technical (97+%) - yellow
liquid partly crystalline
Melting point: base - 32°C HC1 salt - 225 to 227°C with decomposition
Boiling point: base - 163 to 1&5°C at ^k mm Hg
Solubility: base - 250 ppm in water at 20°C; more than 20% in acetone,
benzene, chloroform, ethyl acetate, hexane, methanol
HC1 salt - more than 50% in water, more than 30% in methanol,
1 to 2 % in chloroform, 0.1% in benzene or hexane
Stability: Chlordimeform is hydrolyzed in neutral and acidic media first to
N-formylchlorotoluidine then to ^-chlorotoluidine; very slowly
hydrolyzed in acid media but forms salts; a 0.5% solution of the
HC1 (pH 3 to 4) is stable for some days at 20°C.
-------
3rd Update - August 1982 Chlordimeform EPA-1
Other names: Acaron; Bermat; C 8514; chlorfenamidine (former name); Fundal;
Fundex; Galecron; SN 36268; Spanone; N-(2-methyl-4-
chlorophenyl)-N',N'-dimethyl formamidine; N,N-dimethyl-N'-
(2-methyl-4-chloropheny1)-formamidine
Reagents;
1. Chlordimeform standard of known purity
2. gamma BHC internal standard of known purity
3, Carbon disulfide, ACS grade or better
k. Internal standard solution - weigh 2.5 gram gamma BHC into a 250 ml
volumetric flask, dissolve in and make to volume with carbon disulfide.
(cone 10 mg/ml )
Equipment;
1. Gas chromatograph with flame ionization detector
2. Column: 6" x k mm ID glass packed with a 1:1 mixture of 10% DC-200 and
15% QF-1 on Gas Chrom Q.
3. Precision liquid syringe
k. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions for FID;
Column temperature: 185°C
Injection port temperature: 220°C
Detector temperature: 300°C
Carrier gas: nitrogen - flow adjusted as necessary
Hydrogen flow: adjusted as necessary
Air flow: adjusted as necessary
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibiIity.
Procedure:
Preparation of standard;
Weigh 90 mg Chlordimeform into a small flask or bottle, add 25 ml of internal
standard solution by pipette, stopper tightly, and mix well, (cone 3.6 mg
Chlordimeform and 10 mg gamma BHC per ml)
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3rd Update - August 1982 Chlordimeform EPA-1
Preparation of sample;
Weigh a portion of sample equivalent to 90 mg chlordimeform into a small flask
or bottle, add 25 ml of internal standard solution by pipette, stopper
tightly, and mix well for liquids. For solid samples, shake on a mechanical
shaker for 30 minutes; allow to settle, centrifuge or filter a portion if
necessary taking precautions to avoid evaporation, (cone 3.6 mg chlordimeform
and 10 mg gamma BHC per ml)
GC Determination!
Inject several ul (method submitted did not give injection volume) of standard
and, if necessary, adjust the instrument parameters and the volume injected to
give complete separation within a reasonable time and to obtain peak heights
of 1/2 to 3A full scale. Proceed with the determination, making at least
three injections each of standard and sample solutions. Elution order was not
specified.
Calculation!
Measure the peak heights or areas of the chlordimeform and gamma BHC for both
the standard and sample solutions and calculate the following ratios:
peak height or area chlordimeform
Ratio standard =
Ratio sample =
peak height or area gamma BHC
peak height or area chlordimeform
peak height or area gamma BHC
Average the standard and sample ratios, and calculate the percent
chlordimeform as follows:
(ratio sample) (weight standard) (% purity standard)
~ (ratio standard)(weight sample)
Method submitted by Mississippi State Chemical Laboratory, Mississippi State,
Mississippi 39762
date: unknown but around 1975
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
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3rd Update - August 1982
Chloroneb EPA-1
Determination of Chloroneb by Ultraviolet Spectroscopy
Chloroneb is the accepted (ANSI, BSI, ISO) common name for 1^-dichloro-
2,5-dimethoxybenzene, a registered fungicide having the chemical structure:
OCR.
Molecular formula:
Molecular weight: 207.1
Physical state-color-odor: white crystalline solid with a musty odor
Melting point: 133 to 135°C
Boiling point: 268°C
Solubility: 8 ppm in water at 25°C; soluble in most common solvents:
methylene chloride - 13.3%, dimethyl formamide - 11.8%,
acetone - 11.5%, xylene - 8.9%
Stability: stable at temperatures up to boiling point; stable in common
solvents and in the presence of dilute acid or alkali; subject to
microbial decomposition in moist soil.
Other names: Demosan; Tersan SP
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3rd Update - August 1982 Chloroneb EPA-1
Reagents;
1. Chloroneb standard of known purity
2. Methanol, pesticide or spectro grade
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording with matched
1 cm eel Is
2. Mechanical shaker
3. Centrifuge or filtration apparatus
k. Usual laboratory glassware
Procedure;
Preparation of standard;
Weigh 100 mg chloroneb standard into a 100 ml volumetric flask, dissolve in
and make to volume with methanol; mix thoroughly. Pipette a 10 ml aliquot
into a second 100 ml volumetric flask, make to volume with methanol and mix
thoroughly. Pipette a 25 ml aliquot into a third 100 ml volumetric flask and
make to volume with methanol; mix thoroughly, (final cone 25 ug/ml)
Preparation of sample;
For granular formulations, grind a portion of sample to a fine powder with a
mortar and pestle. Weigh a portion of ground sample equivalent to 100 mg
chloroneb into a 300 ml glass-stoppered flask or screw-cap bottle, add 100 ml
methanol by pipette, close tightly, and shake on a mechanical shaker for one
hour. Allow to settle, centrifuge or filter if necessary, taking precautions
to avoid evaporation of solvent. Dilute 10 ml to 100 ml and then 25 ml to
100 ml as above (second and third flasks) to give final concentration of
25 ug/ml .
UV Determination;
With the UV spectrophotometer at the optimum quantitative settings for the
particular instrument being used, balance the pen at 0 and 100? transmission
at 296 nm with methanol in each cell. Scan both standard and sample solutions
from 330 to 230 nm with methanol in the reference cell. Measure the
absorbance of standard and sample solutions at 296 nm using a reference point
at 330 nm.
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3rd Update - August 1982 Chloroneb EPA-1
CalculatIons;
From the above absorbances and using the standard and sample concentrations,
calculate the percent chloroneb as follows:
(abs. sample) (cone. std. in ug/ml) (% purity of std.)
(abs. std.)(cone, sample in ug/ml)
There is a straight line relationship between absorbance and concentration for
up to 60 ug/ml.
Method submitted by EPA (former) Product Analysis Laboratory, Region II,
New York, NY
January 1977
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
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3rd Update - August 1982
Chlorophacinone EPA-2
Determination of Chlorophacinone by High Performance Liquid Chromatography
Chlorophacinone is the accepted (BSI, ISO) common name for
2-[(p-chlorophenyl)phenylacetyl]-1,3-indandione, a registered rodenticide
having the chemical structure:
0
0
Molecular formula:
Molecular weight: 37^.8
Physical state-color-odor: odorless, white crystalline solid
Melting point:
Solubility: sparingly soluble in water; soluble in organic solvents such as
acetone, ethanol , ethyl acetate
Stability: stable and resistant to weathering; non-corrosive; compatible with
cereals, fruits, roots, and other rodenticide baits; oxidized in
bait formulations
Other names: Caid; Drat; Liphadione; LM 91; Microzul; Ramucide; Ratomet;
Raviac; Rozol; Topitox
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3rd Update - August 1982 Chlorophacinone EPA-2
Reagents;
1. Chlorophacinone standard of known purity
2. Methanol/PIC A - (1 bottle PIC A in one liter of 90% methanol + ^Q% Water
filtered through a 0.^5 micron filter)
3. Water/PIC A - (1 bottle PIC A in one liter of water filtered through a
0.^5 micron f iIter)
Equipment;
1. High Performance Liquid Chromatograph with a variable wavelength UV
detector at 280 nm. If a variable wavelength detector is not available,
operating parameters and concentrations may have to be changed to obtain
the necessary separation and sensitivity.
2. Column: "column A" - uBondapak C18 (30 cm x 3-9 mm ID)
"column B" - Radial Pak C18
3. High pressure liquid syringe or sample injection loop: 5 or 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions:
Mobile phase: "column A" - 72£(90$ methanol/10% water/PIC A) + 28%(water/PIC A)
"column B" - 80£(90S methanol/10? water/PIC A) + 20%(water/P1C A)
Column temperature: "column A" - ambient
"column B" - 32°C
Flow rate: "column A" - 1.0 to 1.5 ml/min
"column B" - 6 ml/min
Wavelength: 280 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg Chlorophacinone standard into a 100 ml volumetric flask, dissolve
in and make to volume with the methanol/PIC A regeant; mix well.
(cone 1 mg/ml)
For "column A" - make a first dilution of 1:10 (5 ml to 50 ml) and a second
dilution of 1:10 (5 ml to 50 ml) using the methanol/PIC A reagent, (final
cone 0.01 mg/ml)
For "column B" - make a dilution of 6 ml to 100 ml with the methanol/PIC A
reagent, (final cone 0.06 mg.ml)
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3rd Update - August 1982 Chlorophacinone EPA-2
Preparation of sample;
Weigh a portion of sample equivalent to 100 mg chlorophacinone into a 125 ml
screw-cap flask, add 100 ml methanol/PIC A reagent by pipette, close tightly,
and shake on a mechanical shaker For one hour.
For "column A" - make the same dilution as above, (cone 0.01 mg/ml)
For "column B" - make the same dilution as above, (cone 0.06 mg/ml)
HPLC Determination!
Filter all solution through a O.Vj micron filter before injecting into the
HPLC. Inject 5 ul (Column A) or 10 ul (Column B) of standard solution and, if
necessary, adjust the flow rate and/or mobile phase composition to give good
separation in a reasonable time. Adjust the attenuation or the amount
injected to give convenient size peaks. Proceed with the determination making
alternately three injections each of standard and sample solutions.
Calculat ion;
Measure the peak height or peak area for each peak and calculate the average
for both standard and sample. Using these averages, calculate the percent
chlorophacinone as follows:
(peak height or area sample)(weight standard injected)(& purity standard)
(peak height or area standard) (weight sample injected)
This method is a combination of a method by Mark W. Law, EPA, Beltsville, MD
dating back to August 1979 and a modification (Radial Pak column) submitted by
Phil Gee and G. Thomas Gale, EPA - NEIC, Denver, CO dated April, 1980.
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Chlorophacinone EPA-3
Determination of Chlorophacinone in Wax Block Baits
by Ultraviolet Spectroscopy
Chlorophacinone is the accepted (BSI, ISO) common name for 2-[(p-
chlorophenyl) phenylacetyl]-1,3-indandione, a registered rodenticide having
the chemical structure:
0
Molecular formula:
Molecular weight: 374.6
Physical state-color-odor: odorless, white crystalline solid
Melting point:
Solubility: sparingly soluble in water; soluble in organic solvents such as
acetone, ethanol, ethyl acetate
Stability: stable and resistant to weathering; non-corrosive; compatible with
cereals, fruits, roots, and other rodenticide baits; oxidized in
bait formulations
Other names: Caid; Drat; Liphadione; LN 91; Microzul; Ramucide; Ratomet;
Raviac; Rozol; Topitox
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3rd Update - August 1982 Chlorophacinone EPA-3
Reagents;
1. Chlorophacinone standard of known purity
2. Benzene, reagent grade
3. Carbon disulfide, reagent grade
k. Ethanol, USP grade 35%
5- Ethanol adjusted to pH 0.5 (on pH meter) with concentrated hydrochloric
ac i d
6. Hexane, reagent grade
7. Florisil 60/100 mesh (regular - not high performance)
Equipment;
1. Ultraviolet spectrophometer, double beam ratio recording with matched
1 cm eel Is
2. Mechanical shaker
3. pH meter
4. Glass chromatographic columns, 1" diameter by 8" (minimum) tall
5. Steambath
6. Usual laboratory glassware
Procedure;
In this method, a standard, sample, and fortified sample are run parallel to
each other. A percent recovery is determined and used to calculate the
corrected percent Chlorophacinone from the percent found in the determination.
Preparation of standard;
For a stock standard solution - weigh 50 mg Chlorophacinone into a 100 ml
volumetric flask, dissolve in and make to volume with benzene; mix well.
(cone 0.5 mg/ml) Use one ml of this solution for the determination - see
column clean-up below.
Preparation of sample;
Weigh a portion of sample equivalent to 0.5 mg Chlorophacinone (10 grams for a
0.005? formulation) into a 250 ml screw-cap flask, add 100 ml benzene by
graduated cylinder, close tightly, and shake on a mechanical shaker for one
hour. Decant the extract through Whatman #k (or equivalent) filter paper,
collecting the filtrate in a *tOO ml beaker. Add 50 ml benzene to the flask
and shake for 30 minutes; decant through the same filter collecting the
filtrate in the same AGO ml beaker. Add another 50 ml benzene, shake again
for 30 minutes, and decant through the same filter as above. Use the combined
extracts for the determination - see column clean-up below.
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3rd Update - August 1982 Chlorophacinone EPA-3
Preparation of fortified sample;
Weigh another portion of sample as above but add 0.5 ml of stock standard
solution (0.25 mg Chlorophacinone) and extract exactly as above. Use the
combined extracts for the determination - see column clean-up below.
Column clean-up;
Pack three 1" diameter x 8" length glass columns with 20 grams (each) of
regular florisil; wet with benzene.
Quantitatively place the standard, sample, and fortified sample onto the
pre-wetted columns with benzene. Wash each column with 300 ml benzene
followed by 100 ml carbon disulfide and 100 ml nexane. Discard the
washings. Elute the Chlorophacinone from the columns with 130 ml of the 0.5
pH EtOH/HC1 solution. Collect the eluates in 250 ml beakers. Concentrate on
a steambath to approximately 75 ml and quantitatively transfer to 100 ml
volumetric flasks and make to volume with 35% ethanol . Filter 10 to 20 ml
through Whatman #3 (or equivalent) filter paper. Use the filtered solutions
for UV determination.
UV Determination;
With the UV spectrophotometer at the optimum quantitative settings for the
particular instrument being used, balance the pen at 0 and 100% transmission
at 360 nm with 0.5 pH EtOH/HC1 solution in each cell. Scan the standard,
sample, and fortified sample solutions from 240 to 360 nm with 0.5pH EtOH/HC1
solution in the reference cell. Measure the absorbance at 321 nm and 350 nm
for each solution.
Cal cul at ions;
From the above absorbances and the weights of standard, sample, and fortified
sample, make the following calculations:
(abs ,~ abs-3c samP'e) (m9 standard)
mg sample found =
,— - - - — - - — -^ -
(abs, 21 - abs,rgStandard)
(abs 321" a^s350 fort'fied sample) (mg standard)
mg fortified sample found
(abs,~ abs,j-gStandard)
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3rd Update - August 1982 Chlorophacinone EPA-3
(mq fortified sample found) - (mq sample found) (100)
* r ec ov e ry =
(mg Chlorophacinone added for fortification)
(abs 021" aks7<;n sample) (mg standard) (100)
% Chlorophacinone =
% corrected Chlorophacinone
Notes:
jStandard) (mg sample)
(% Chlorophacinone found) (100)
T$ recovery)
1. All glassware must be cleaned with Pierce PBS-35 concentrate (dilute:
AO ml per 1000 ml water) or equivalent. Otherwise interferring substances
remain on the glassware and interfere at the following UV analysis.
2. Since there is concern over the use of benzene in the laboratory, the
amount used can be decreased by:
(1) decreasing the sample size to 5 gram (0.005% formulation) and
extracting with 50 ml, 25 ml, 25 ml portions
(2) using only enough benzene to wash the paraffin from the column (less
than 300 ml) until no more paraffin can be detected
(3) once the recovery of a particular sample has been determined to be
very good, analysis of a fortified sample can be omitted during
repeat analyses.
Method submitted by University of Hawaii at Manoa, Department of Agricultural
Biochemistry, Honolulu, Hawaii (Wanda L. Chang and Y. Kawano)
October 1979
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982 Dicrotophos EPA-1
Determination of Dicrotophos by Infrared Spectroscopy
Dicrotophos is the accepted (BSI, ISO) common name for 3~
(dimethoxyphosphinyloxy)-N,N-dimethyl-cis-crotonamide, a regi stered
insecticide having the chemical structure:
CH.CX ° H °
XJ I II
\p—o—c=c—-c—>
I
CHo
Molecular formula: CnH.gOVPN
Molecular weight: 237-2
Physical state-color-odor: yellow to brown liquid with a mild ester odor
Boiling point: 400°C
Solubility: miscible with water, acetone, methanol, ethanol, isopropanol,
xylene; very slightly soluble in kerosene and diesel fuel
Stability: stable up to 40 C when stored in glass or polyethylene containers;
decomposes after 31 days at 75°C or 7 days at 90°C. The half-life
of an aqueous solution at 38°C and pH 9.1 is 1200 hours, at pH 1.1
it is 2400 hours. Formulations on most carriers are unstable;
acidic solutions are more stable than basic solutions. Compatible
with most other pesticides. Relatively non-corrosive to Monel,
copper, nickel, and aluminum; somewhat corrosive to cast iron,
mild steel, brass, and stainless steel 304; does not attack glass,
polyethylene, or stainless steel 316.
Other names: Bidrin; C709; Carbicron, Ektafos; SD 3562
Reagents^
1. Dicrotophos standard of known purity
2. Carbon tetrachloride, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
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3rd Update - August 1982 Dicrotophos EPA-1
Equipment;
1. Infrared spectrophotometer, double beam, with matched 0.5 mm NaC1 or KBr
eel Is
2. Mechanical shaker
3. Centrifuge or filtration apparatus
k. Usual laboratory glassware
Procedure:
Preparation of standard;
Weigh 90 mg dicrotophos standard into a 10 ml volumetric flask, dissolve in
and make to volume with carbon tetrachloride. Add a little anhydrous sodium
sulfate to insure dryness. (cone 9 mg/ml)
Preparation of sample;
For 1i quids - weigh a portion of sample equivalent to 900 mg dicrotophos into
a 100 ml volumetric flask, make to volume with carbon tetrachloride, and mix
thoroughly. Add a little anhydrous sodium sulfate to insure dryness.
For dusts, granules, and wettable powders - weigh a portion of sample
equivalent to 900 mg dicrotophos into a 250 - 300 ml glass-stoppered flask or
screw-cap bottle, add 100 ml carbon tetrachloride by pipette and some
anhydrous sodium sulfate, close tightly, and shake on a mechanical shaker for
one to two hours. Allow to settle, centrifuge or filter if necessary, taking
precautions to prevent evaporation of solvent, (cone 9 mg/ml)
IR Determinat ion:
With carbon tetrachloride in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument being used,
scan both the standard and sample solutions from 960 to 880 cm (10.4 to
11.4 urn). Determine the absorbance of standard and sample using the peak at
926 cnf1 10.8 urn) and a reference point at 901 cm""'(11.1 urn).
Calculation;
From the above absorbances and using the standard and sample concentrations,
calculate the percent dicrotophos as follows:
% = (aks' sample) (cone, std. in mg/ml) (% purity of std.)
(abs. std.) (cone, sample in mg/ml)
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3rd Update - August 1982 Dicrotophos EPA-1
This method was submitted by the Mississippi State Chemistry Laboratory
sometime around 1975- It was submitted as a basic outline and not a fully
written procedure. I have taken the liberty of writing it in our standard IR
format.
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Dimethyl phthalate EPA-1
Determination of Dimethyl phthalate by Gas Chroma tography (FID-IS)
Dimethyl phthalate (common name and chemical name) is a registered insect
repellent having the chemical structure:
• CH,
Molecular formula:
Molecular weight: 19^.2
Physical state-color-odor: colorless to faintly yellow viscous liquid,
slight aromatic odor
Boiling point: 282 to 285°C
Solubility: practically insoluble in water (0.^3 g/100 ml), petroleum ether,
and other paraffin hydrocarbons; miscible with alcohol, ether,
chloroform, and most organic liquids
Stability: stable, though hydrolyzed by alkalis
Other names: DMP; 1,2-benzenedicarboxylic acid dimethyl ester; dimethyl
1,2-benzenedicarboxylate
Note: This method was developed for "mosquito cloth wipes" but can easily be
adapted for other types of formulations.
Reagents;
1. Dimethyl phthalate standard of known purity
2. Pentachloronitrobenzene, purified or analytical grade
-------
3rd Update - August 1982 Dimethyl phthalate EPA-1
3. Acetone, pesticide or analytical grade
k. Internal standard solution - weigh 4.0 grams pentachloronitrobenzene into
a 500 ml volumetric flask, dissolve in and make to volume with acetone;
mix thoroughly, (cone 8 mg/ml)
Equipment:
1. Gas chromatograph with flame ionization detector
2. Column: 6' x 1/8" stainless steel packed with 3% XE-60 on 80 to 100 mesh
Chromosorb W (or equivalent column)
3. Precision liquid syringe: 10 ul
k. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions for FID;
Column temperature: 160°C
Injection port temperature: 220°C
Detector temperature: 250°C
Carrier gas: nitrogen - flow (adjusted as necessary)
Hydrogen flow: (adjusted as necessary)
Air flow: (Adjusted as necessary)
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure;
Preparation of standard;
Weigh 400 mg dimethyl phthalate standard into a screw-cap flask or bottle and
add 100 ml internal standard solution by pipette; mix thoroughly.
(cone 4 mg dimethyl phthalate and 8 mg pentachloronitrobenzene per ml)
Preparation of sample;
Cut small squares from different areas of cloth to obtain a representative
sample. Weigh a portion of the cut up sample equivalent to 400 mg dimethyl
phthalate into a 250 ml screw-cap flask and add 100 ml internal standard
solution by pipette. Shake on a mechanical shaker for 30 minutes. Filter or
centrifuge, as necessary, taking precaution to avoid evaporation of the
acetone, (cone 4 mg dimethyl phthalate and 8 mg pentachloronitrobenzene
per ml)
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3rd Update - August 1982 Dimethyl phthalate EPA-1
GC Determination;
Inject 2 ul of standard and, if necessary, adjust the instrument parameters
and the volume injected to give a complete separation within a reasonable time
and to obtain peak heights of 1/2 to 3A full scale. Proceed with the
determination, making at least three injections each of standard and sample
solutions. The elution order is dimethyl phthalate then pentachloronitrobenzene.
Calculat ion;
Measure the peak heights or areas of the dimethyl phthalate and pentachloro-
nitrobenzene for both the standard and sample solutions and calculate the
following ratios:
peak height or area dimethyl phthalate
Ratio of standard
Ratio of sample =
peak height or area pentachloronitrobenzene
peak height or area dimethyl phthalate
peak height or area pentachloronitrobenzene
Average the standard and sample ratios, and calculate the percent dimethyl
phthalate as follows:
(ratio of sample) (weight of standard) (% purity of standard)
(ratio of standard) (weight of sample)
Method submitted by E. S. Greer, EPA (formerly) Product Analysis Laboratory.
Region IX, San Francisco, CA (Mr. Greer is now at Beltsville, MD)
February 1976
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Din itramine EPA-1
Determination of Dinitramine by Gas Chromatography (FID-IS)
Dinitramine is the accepted (BSI, ISO, WSSA) common name for N*,N -
diethyl-a,a,a-trif1uoro-3,5-dinitrotoluene-2,4-diamine, a registered herbicide
having the chemical structure:
Molecular formula: C... H..,F,N. 0.
Molecular weight: 322.2
Physical state-color-odor: yellow crystal 1ine solid
Melting point: 98 to 99°C
Solubility: grams per 100 ml solvent at 20°C; acetone - 10**, benzene - '+7.3,
ethanol - 10.7, chloroform - 67, hexane - l.k, xylene - 22.7,
water - 0.0001 (1 ppm)
Stability: relatively stable at room temperature; decomposes above 200°C;
subject to photodegradation; non-corrosive
Other names: Cobex, Cobexo; USB 3584
Reagents^;
1. Dinitramine standard of known purity
2. Dibutyl phthalate (internal standard), (Dibutyl phthalate of known purity)
-------
3rd Update - August 1982 Dinitramine EPA-1
3. Acetone, pesticide grade
!*. Internal standard solution - weigh 1.0 gram dibutyl phthalate into a
100 ml volumetric flask, dissolve in and make to volume with acetone; mix
well, (cone 10 mg/ml)
Eg u i pmen t;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: V x 1/V glass packed with k% SE-30 on 80 to 100 mesh Chromosorb
W HP (or equivalent column)
3. Precision liquid syringe: 10 ul
4. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions for FID;
Column temperature: 185°C
Injection port temperature: 200°C
Detector temperature: 235°C
Carrier gas: Helium (or nitrogen) - 30 ml/min (adjusted as necessary)
Hydrogen flow: (adjusted as necessary)
Air flow: (adjusted as necessary)
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 110 mg dinitramine standard into a 100 ml volumetric flask, add 10 ml
internal standard solution by pipette, make to volume with acetone, and mix
thoroughly, (cone 1.1 mg dinitramine and 1 mg dibutyl phthalate per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 110 mg dinitramine into a 100 mi
volumetric flask, add 10 ml internal standard solution by pipette, make to
volume with acetone, and mix thoroughly, (cone 1.1 mg dinitramine and 1 mg
dibutyl phthalate per ml)
If formulations other than liquid are encountered, proceed as follows:
weigh a portion of sample equivalent to 110 mg dinitramine into a 250 ml
glass-stoppered flask or screw-cap bottle, add 10 ml internal standard
-------
3rd Update - August 1982 Dinitramine EPA-1
solution and 90 ml of acetone by pipette, stopper tightly and shake on a
mechanical shaker for one hour. Allow to settle, centrifuge or filter if
necessary, taking precautions to avoid evaporation of the acetone.
(Cone as above)
GC Determinat ion;
Inject 3 to k ul of standard and, if necessary, adjust the instrument
parameters and the volume injected to give a complete separation within a
reasonable time and to obtain peak heights of 1/2 to 3A full scale. Proceed
with the determination, making at least three injections each of standard and
sample solutions. The elution order is dinitramine then dibutyl phthalate.
Calculat ion;
Measure the peak heights or areas of the dinitramine and dibutyl phthalate for
both the standard and sample solutions and calculate the following ratios:
peak height or area dinitramine
Ratio standard =-
peak height or area dibutyl phthalate
peak height or area dinitramine
Ratio sample =
peak height or area dibutyl phthalate
Average the standard and sample ratios, and calculate the percent dinitramine
as follows:
(ratio sample) (weight standard) (% purity standard)
(ration standard) (weight sample)
Method submitted by EPA (former) Product Analysis Laboratory, Region II,
New York, NY
March 1977
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
DIoxins EPA-1
Determination of DIoxins at the Parts Per Billion Level
in Technical and Formulated 2,4-D and 2,1*,5-T Using GC/MS
Chlorinated dibenzo-p-dioxins occur as contaminants in 2,4-D and 2,4,5-T
herbicides. These dioxins are formed when chlorinated benzenes are treated
with high temperature and pressure under alkaline conditions such as are used
in the manufacture of chlorophenoxy acid herbicides. Although they are by-
products of the manufacturing process, they are considered contaminants. Some
of these materials are known to be extremely toxic to test animals, therefore,
they are highly undesireable in the environment.
Polychlorinated dioxins are formed by chlorination of dibenzo-p-dioxin
which has the structural formula:
The substitution of from one to eight chlorines takes place at the carbon
atoms numbered 1,2,3,^,6,7,8,9 en the above structure. These polychlorinated
dioxins are sparingly soluble in most organic solvents and have limited
solubility in water. The molecular weights range from 185 for no chlorine
to ^60 for eight chlorines (C.
(C12Hg02)
,12ci8o2).
This method will identify all 8 dioxin species at the parts per billion
level. The low resolution GC/MS procedure has a sensitivity of about 1 ppb
for most isomers but will not separate all known dioxins.
Reagents;
1. Dioxin standards - available from Altech Inc.
2. Silica gel , MCB grade 923
3. Alumina, Woelm B, Super 1 used as is
^. Acetonitrile, pesticide grade
5. Carbon tetrachloride, pesticide grade
6. Hexane, pesticide grade
7. Methyl Alcohol, pesticide grade
8. Methylene chloride, pesticide grade
9. Sodium sulfate, anhydrous, granular
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3rd Update - August 1982 Dioxins EPA-1
10. Sodium hydroxide - 1 N solution
11. Sulfuric acid - 1 N solution and concentrated
12. 1+1 acetonitrile water (with 10% methyl alcohol)
13. 1+1 methyl alcohol/water
jig u i pmen t;
1. Low resolution GC/MS caple of single ion monitoring. The sensitivity of
the instrument should be at least 1 ng for any given dioxin standard when
monitoring a single ion.
2. GC column for above: 6' x 2 MM ID glass packed with 1.5% OV-101 on 80/100
Chromosorb W HP (or equivalent column)
3. Precision microliter syringes (Hamilton or equivalent)
k. Usual laboratory glassware
Operation conditions;
Column temperature: 200 to 250°C depending on the dioxin species for which
the analyst is being done.
All other parameters should be those giving the best conditions for the
particular instrument being used.
Procedure;
Technical 2,^-D and 2,4,5-T acids;
Dissolve 10 gram of acid in 400 ml of 1 + 1 acetonitr i le/water (with 10%
methyl alcohol) solution. When totally dissolved, transfer to a 1000 ml
separatory funnel and extract with 3 x 100 ml portions of hexane. Combine the
hexane extracts in another separatory funnel.
Wash the combined hexane extracts as follows, discarding each wash solution:
k x 100 ml portions of 1 + 1 methyl alcohol/water
3 x 100 ml portions of 1 N NaOH solution
3 x 100 ml portions of 1 N H^O^ solution
3 x 100 ml portions of water
Filter the hexane through sodium sulfate and evaporate with dry nitrogen to
about 25 ml . Shake the hexane with 4 x 25 ml portions of concentrated H2SQ/,
and discard the acid. Wash with 2 x 25 ml portions of water and discard the
water. Filter through sodium sulfate and evaporate to about 5 to 10 ml using
dry nitrogen.
Prepare alumina column as follows: 3" Wohlem alumina topped with 1" sodium
sulfate: wash with 200 ml hexane and discard the hexane.
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3rd Update - August 1982 Dioxins EPA-1
Transfer the evaporated hexane sample solution to the column with a minimum of
hexane. Elute with 100 nl hexane and discard the eluate. Eluate with 2%
methylene chloride in hexane and discard. Elute with 200 ml 30$ methylene
chloride in hexane and save the eluate. Evaporate with dry nitrogen to 0.5 to
1.0 ml for GC/MS.
Esters and Formulations;
Set up silica gel column using 30 grams in a 2 cm x 60 cm column topped with
sodium sulfate. Transfer 2 to 5 grams sample fco column and eluate with 200 ml
30% methylene chloride in hexane. Follow the above procedure beginning with
the paragraph "Wash the combined hexane extracts "
Interferences from PCBs:
If PCBs are suspected of interfering with the analysis, they can be eliminated
as follows. Prepare a 30 cm x 12 mm ID chromatographic column by adding
1 1/2" of alumina and topping with 1" of anhydrous sodium sulfate. Wash
column with methylene chloride and purge with a stream of dry nitrogen.
Activate column at 225°C for approximately 6 hours and store at 125 C until
ready to use.
Prewet column with hexane and transfer 1 ml sample solution to column using
two 1 ml portions of hexane. Elute sample with 20 ml of carbon tetrachloride
and discard eluate. Elute with 15 ml of methylene chloride. Collect eluate
and evaporate to just dryness with a stream of nitrogen. Dilute to 0.5 to
1.0ml for analysis.
Determinat ion;
For GC/MS, inject aliquots of the sample extract and appropriate standards
into the instrument which has been tuned to maximum efficiency and sensitivity
for dioxins.
The ions to be monitored for the 8 dioxin species are;
monochloro 218 amu
dichloro 252 amu
trichloro 288 amu
tetrachloro 322 amu
pentachloro 356 amu
hexachloro 391 amu
heptachloro ^26 amu
octachloro *»60 amu
Method submitted by EPA Beltsville Chemistry Laboratory (Ronald F. Thomas and
Everett S. Greer) Beltsville, MD April 1982
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Diphacinone EPA-3
Determination of Diphacinone by High Performance Liquid Chromatography
Diphacinone is the accepted common name for 2-{diphenylacetyl)-1,3-
indandione, a registered rodenticide having the chemical structure:
o
II
-c-
• C—H
Molecular formula:
Molecular weight:
Physical state-color-odor: odorless, yellow crystals
Melting point: 145°C
Solubility: slightly soluble in water and benzene; soluble in acetone and
acetic acid; forms a sodium salt which is sparingly soluble in
water
Stability: resists hydrolysis; stable toward mild oxidants; non-corrosive
Other names: diphacin (Turkey); Diphacin; Diphacin Meal Bait; P.C.Q.; Promar;
Ramik; Rodent cake
Reagents;
1. Diphacinone standard of known purity
2. Methanol/PIC A - (1 bottle PIC A in one liter of 30% methanol/10& water
filtered through a 0.45 micron filter)
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3rd Update - August 1982 Diphacinone EPA-3
3. Water/PIC A - (1 bottle PIC A In one liter of water filtered through a
0.1*5 micron f i Iter
Equipment;
1. High Performance Liquid Chromatograph with a variable wavelength UV
detector at 312 nm. If a variable wavelength detector is not available,
operating parameters and concentrations may have to be changed to obtain
the necessary separation and sensitivity.
2. Column: MCH-10 (30 cm x 4 mm ID)
3. High pressure liquid syringe or sample injection loop: 10 ul
A. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 65% (90fc methanol/10fc water/PIC A) = 35% (water/PIC A)
Column temperature: 32°C
Flow rate: 1.5 ml/min
Wavelength: 312 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi15ty.
Procedure:
Preparation of standard;
Weigh 60 mg diphacinone standard into a 100 ml volumetric flask, dissolve in
and make to volume with the methanol/PIC A solution. Dilute 5 ml to 50 ml
with this same solution. Filter a portion through a 0.*»5 micron filter.
(cone 0.06 mg/ml)
Preparation of sample;
Weigh a portion of sample equivalent to 60 mg diphacinone into a 100 ml
volumetric flask, make to volume with the methanol/PIC A solution, and mix
thoroughly. Dilute 5 ml to 50 ml with this same solution. Filter a portion
through a 0.^5 micron filter, (cone as above)
HPLC Determination;
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give a good separation in a reasonable
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3rd Update - August 1982 Diphacinone EPA-3
time. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample solutions.
Calculation;
Measure the peak height or peak area for each peak and calculate the average
for both standard and sample. Using these averages, calculate the percent
diphacinone as follows:
(peak height or area sample)(weight standard injected)(% purity standard)
(peak height or area standard)(weight sample injected)
Method submitted by EPA - NEIC, Denver, Colorado (Phil Gee & G. Thomas Gale)
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
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3rd Update - August 1982
Diphenamid EPA-1
Determination of Diphenamid by Infrared Spectroscopy
Diphenamid is the accepted (ANSI, BSI, ISO, MAPJ, WSSA) common name for
N,N-dimethyl-2,2-diphenylacetamide, a registered herbicide having the chemical
structure:
CH,
Molecular formula: C^H^NO
Molecular weight: 239-3
Physical state-color-odor: white or off-white crystalline solid, has no
appreciable odor
Melting point: 13^-5 to 135-5°C (pure); 128 to 135°C (35% technical)
Solubility: grams per 100 ml solvent at 27°C: acetone - 19, dimethyl
formamide - 16.5, phenvl cellosolve - 32, xylene - 5,
water - 0.026 (260 ppm)
Stability: moderately stable to heat and UV light; compatible with most
other pesticides; non-corrosive, non-flammable
Other names: Dymid; Enide; L-
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3rd Update - August 1982 Diphenamid EPA-1
Reagents:
1. Diphenamid standard of known purity
2. Carbon disulfide, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
Equipment:
1. Infrared spectrophotometer, double beam, with matched 0.5 mm KBr cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4, Usual laboratory glassware
Procedure:
Preparation of standard;
Weigh 150 mg diphenamid standard into a 25 ml volumetric flask, dissolve in
and make to volume with carbon disulfide. Add a little anhydrous sodium
sulfate to insure dryness, and shake well, (cone 6 mg/ml)
Preparation of sample;
1 i quids - weigh a portion of sample equivalent to 600 mg diphenamid into a
100 ml volumetric flask, make to volume with carbon disulide and mix
thoroughly. Add a little anhydrous sodium sulfate to insure dryness and shake
wel 1 . (cone 6 mg/ml )
For dusts; granules, and wettable powders - weigh a portion of sample
equivalent to 600 mg diphenamid into a 250 - 300 glass-stoppered flask or
screw-cap bottle, add 100 ml carbon disulfide by pipette and some anhydrous
sodium sulfate, close tightly, and shake on a mechanical shaker for one
hour. Allow to settle, centrifuge or filter if necessary, taking precautions
to avoid evaporation of solvent, (cone. 6 mg/ml)
IR Determination:
With carbon disulfide in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument being used,
scan both the standard and sample solutions from 7^0 to 665 cm" (13.5
to 15-0 urn). Determine the absorbance of standard and sample using the peak
at 700 cm"1 (ik.3 urn) and a baseline from 725 to 678 cm~1 (13.8 to H.75 urn).
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3rd Update - August 1982 Diphenamid EPA-1
Calculation;
From the above absorbances and using the standard and sample concentrations,
calculate the percent diphenamid as follows:
* _ (abs. sample) (cone, std. in mg/ml) (% purity of std.)
(abs. std.)(cone, sample in mg/ml)
This method has been used several times in the past (back to the middle and
late 1960's) in the Beltsville Laboratory; it seems satisfactory, but has
never been formally checked-out.
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Diphenamid EPA-2
Determination of Diphenamid by Gas Liquid Chromatography (FID-IS)
Diphenamid is the accepted (ANSI, BSI, ISO, MAPJ, WSSA) common name for
N,N-dimethyl-2,2-diphenylacetamide, a registered herbicide having the chemical
structure:
H
Molecular formula: C..gH..,NO
Molecular weight: 239-3
Physical state-color-odor: white or off-white crystalline solid, has no
appreciable odor
Melting point: 13^.5 to 135.5°C (pure); 128 to 135°C (35% technical)
Solubility: grams per 100 ml solvent at 27 C = acetone - 19; dimethyl
formamide - 16.5; phenyl cellosolve - 32; xylene - 5,
water - 0.026 (260 ppm)
Stability: moderately stable to heat and UV light; compatible with most
other pesticides; non-corrosive; non-flammable
Other names: Dymid; Enide; L-
Reagents;
1. Diphenamid standard of known purity
2. HEOD (100%, or dieldrin of known HEOD content) internal standard
(See note #1)
3. Chloroform, pesticide grade
A. Internal standard solution - weigh 3-5 grams HEOD (or dieldrin equivalent
to 3-5 grams HEOD) into a 250 ml volumetric flask, dissolve in and make to
volume with chloroform, and mix well, (cone 1*f mg/ml)
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3rd Update - August 1982 Diphenamid EPA-2
Equipment;
i. Gas chroma tograph with flame ionization detector (FID)
2. Column: 6' x k mm ID glass packed with 3% Poly I (polyethylene imine)
110 on 80 to 100 mesh Gas Chrom Q (or equivalent column - suggest trying
SP-1000 on Chromosorb 750, if available)
3. Precision liquid syringe: 10 ul
A. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions for FID;
Column temperature: 260°C
Injection port temperature: 300°C
Detector temperature: 300°C
Carrier gas: Nitrogen - (flow adjusted as necessary)
Hydrogen flow: (adjusted as necessary)
Air flow: (adjusted as necessary)
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure;
Preparation of standard;
Weigh 200 mg diphenamid standard into a 100 ml volumetric flask, add 50 ml
internal solution by pipette, and make to volume with chloroform; mix well.
(cone 2 mg diphenamid and 7 mg HEOD per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 200 mg diphenamid into a 250 ml glass-
stoppered flask or screw-cap bottle, add 50 ml internal standard by pipette
and 50 ml chloroform by pipette, close tightly, and shake on a mechanical
shaker for about one hour. Allow to settle, centrifuge or filter if
necessary, taking precautions to avoid evaporation of the acetone.
(cone approx. 2 mg diphenamid and 7 fig HEOD per ml)
GC Determination;
Inject 2 ul of standard and, if necessary, adjust the instrument parameters
and the volume injected to give a complete separation within a reasonable time
and to obtain peak heights of 1/2 to 3A full scale. Proceed with the
determination, making at least three injections each of standard and sample
solutions. The elution order is diphenamid then HEOD.
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3rd Update - August 1982 Diphenamid EPA-2
Calculation:
Measure the peak heights or areas of the diphenamid and HEOD for both the
standard and sample solutions and calculate the following ratios:
peak height or area diphenamid
Ratio standard =
peak height or area HEOD
peak height or area diphenamid
Ratio sample = -
peak height or area HEOD
Average the standard and sample ratios, and calculate the percent diphenamid
as follows:
(ratio sample) (weight standard) (% purity standard)
(ratio standard)(weight sample)
Method submitted by Division of Regulatory Services, Lexington, Kentucky
Note #1
Any information as to other internal standards will be appreciated - prefer
not to use pesticides as internal standards
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Diquat EPA-1
Determination of Diquat by High Performance Liquid Chromatography
Diquat is the accepted (BSI, ISO) common name for 1,1'-ethylene-2,2'-
bipyridylium ion; usually present as the dibromide monohydrate salt. Diquat
is a registered herbicide and desiccant and has the chemical (cation)
structure:
2 +
Molecular formula: ^12^12^2 (cat'on)» ^12^1
(dibromide salt)
Molecular weight: 18A.2 (cation); 3M.1 (dibromide salt)
Physical state-color-odor: (dibromide salt) - white to yellow crystals
(aqueous solution) - dark reddish brown
Melting point: decomposes above 300 C, charring rather than melting or
boi1 ing
Solubility: (dibromide) -
very soluble in water (70 grams/100 ml at 20°C);
slightly soluble in alcohol and hydroxyllc
solvents; practically insoluble in non-polar
organic solvents
Stability: (dibromide) - stable in acid or neutral solutions but unstable
under alkaline conditions; may be inactivated by
inert clays and anionic surfactants; decomposes in
UV light; unformulated products are corrosive to
metals
Other names:
deiquat (Germany); region (USSR); Aquacide; Dextrone; Reglone;
Reglox; Weedtrine-D; 6,7-dihydrodipyrido[1,2-a:2',1'-c ]
pyrazinedinium ion
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3rd Update - August 1982 Diquat EPA-1
Reagents:
1. Diquat (dibromide) standard of known purity
2. Phenol (internal standard) of known purity (make sure that the phenol
gives a clean chromatogram with no co-eluting peaks)
3. Aqueous mobile phase - (0.0025M 1-heptane sulfonic acid sodium salt and
0.04M tetramethylammonium chloride adjusted to
pH 3.0 with sulfuric acid) Filter through a 0.45
micron fi1ter.
4. Organic mobile phase - (0.06M tetramethylammonium chloride in 200 ml water
adjusted to pH 3-0 with sulfuric acid plus 800 ml
acetonltrile) Filter through a 0.45 micron filter.
5. Internal standard solution - weigh 1 gram phenol into a 500 ml volumetric
flask, dissolve in and make to volume with aqueous mobile phase.
(cone 2 mg/ml)
Equi pment:
1. High Performance Liquid Chromatograph with UV detector at 254 nm. If a
variable wavelength detector is available, other wavelengths may be used
to increase sensitivity or to eliminate interference.
2. Column: MicroPak MCK-10 (30 cm x A mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 85% aqueous mobile phase + 15% organic mobile phase
Column temperature: ambient
Flow rate: 2 ml/min
Wavelength: 254 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure^
Preparation of standard;
Weigh 50 mg diquat (dibromide) standard into a 100 ml volumetric flask,
dissolve in and make to volume with internal standard solution; mix well.
Filter a portion through a 0.45 micron filter, (cone 0.5 mg diquat
(dibromide) and 2 mg phenol per ml)
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3rd Update - August 1982 Diquat EPA-1
Preparation of sample;
Weigh a portion of sample equivalent to 50 mg diquat (dibromide) into a 125 ml
screw-cap flask, add 100 ml internal standard solution by pipette, close
tightly, and shake on a mechanical shaker for 30 minutes. Filter a portion
through a 0.^5 micron filter, (cone 0.5 rng diquat (dibromide) and 2 mg phenol
per ml)
HPLC Determination:
Inject 10 ul standard solution and, if necessary, adjust the flow rate and/or
mobile phase composition to give good separation in a reasonable time. Adjust
the attenuation or the amount injected to give convenient size peaks. Proceed
with the determination making alternately three injections each of standard
and sample solution.
Calculat ion:
Measure the peak heights or areas of the diquat (dibromide) and the phenol for
both the standard and sample solutions and calculate the following ratios:
peak height or area diquat (dibromide)
Ratio standard y
Ratio sample
peak height or area phenol
peak height or area diquat (dibromide)
peak height or area phenol
Average the standard and sample ratios, and calculate the percent diquat
(dibromide) as follows:
(ratio of sample) (weight of standard) (% purity of standard)
(ratio of sample)(weight of sample)
Method submitted by EPA - NEIC, Denver, Colorado (G. Thomas Gale)
January 1980
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982 Oisulfoton EPA-3
Determination of Disultoton by Gas Chroma tography (FID-IS)
Disulfoton is the accepted (BSI, ISO) common name for 0,0-diethyl
S-[2-(ethylthio)ethyl] phosphorodithioate, a registered insecticide and
acaricide having the chemical structure:
Q— CH0— 0> S
3 2
— S— CH2 — CH2— S— CH2—
Molecular formula:
Molecular weight:
Physical state-color-odor: pure - colorless to pale yellow liquid with a
characteristic odor of sulfur compounds;
technical - dark yellowish oil with an aromatic
odor
Boiling point: 62°C at 0.01 mm Hg
Solubility: 25 ppm in water at room temperature; readily soluble in most
organic solvents
Stability: subject to hydrolysis under alkaline conditions; stable in normal
storage
Other names: Bay 2?6; Bay 19&39; Disyston; Di-Syston (in U.S.);
dithiodemeton; dithiosustox; Frumin AL; M-7^ (USSR); Solvirex
Reagents;
1. Disulfoton standard of known purity
2. Dibutyl phthalate of known purity
3. Acetone, pesticide grade
^. Internal standard solution - weigh 600 mg dibutyl phthalate into a 250 ml
volumetric flask, dissolve in and make to volume with acetone; mix well.
(cone 2.k mg/ml)
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3rd Update - August 1982 Disulfoton EPA-3
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 1/V glass packed with 10% OV-1 on 80 to 100 mesh
Chromosorb W (or equivalent column)
3. Precision liquid syringe: 10 ul
k. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating parameters for FID;
Column temperature: 220°C
Injection port temperature: 250°C
Detector temperature: 250°C
Carrier gas: Nitrogen (flow adjusted as necessary)
Hydrogen flow: (adjusted as necessary)
Air flow: (adjusted as necessary)
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure;
Preparation of standard;
Weigh 100 mg disulfoton standard into 125 ml screw-cap flask, add 50 ml of
internal standard solution by pipette, and shake to dissolve, (cone 2 mg
disulfoton and 2.k mg dibutyl phthalate per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 100 mg disulfoton into a 125 ml screw-
cap flask, add 50 ml internal standard solution by pipette, and shake on a
mechanical shaker for about one hour. Allow to settle, centrifuge or filter
if necessary, taking precaution to avoid evaporation of the acetone, (cone
approx. 2 mg disulfoton and 2.A rng dibutyl phthalate per ml)
GC Determinat ion;
Inject 2 to 4 ul of standard and, if necessary, adjust the instrument
parameters and the volume injected to give a complete separation within a
reasonable time and to obtain peak heights of 1/2 to 3/4 full scale. Proceed
with the determination, making at least three injections each of standard and
sample solutions. The elution order is disulfoton then dibutyl phthalate.
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3rd Update - August 1982 Disulfoton EPA-3
Calculat ion;
Measure the peak heights or areas of the disulfoton and dibutyl phthalate for
both the standard and sample solutions and calculate the following ratios:
peak height or area disulfoton
Ratio standard =
peak height or area dibutyl phthalate
peak height or area disulfoton
Ratio sample = •—-—— — —•———
peak height or area dibutyl phthalate
Average the standard and sample ratios, and calculate the percent disulfoton
as follows:
(ratio sample) (weight standard) (% purity standard)
(ratio standard)(weight sample)
Method submitted by E. S. Greer, August 1977, EPA (formerly) Product Analysis
Laboratory, San Francisco, CA and presently Beltsville Chemical Laboratory,
Beltsvilie, MD
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982 Epichlorohydrin EPA
Determination of Epichlorohydrin by Gas Chromatography (FID)
Epichlorohydrin is the classical common name for 1-chloro-2,3-
epoxypropane, a registered insect fumigant having the chemical structure:
,0.
Molecular formula: C-,H,-C10
Molecular weight: 92.53
Physical state-color-odor: very volatile, narcotic liquid, with a chloroform-
1 ike odor
Melting point: -25.6°C
Boiling point: 115 to 117°C
Solubility: misciblewith most organic solvents; immiscible with water and
petroleum hydrocarbons
Stability: unstable
Other names: chloropropylene oxide; chloromethyloxirane
Reagents;
1. Epichlorohydrin standard of known purity
2. Acetone, ACS grade (other solvents may be used if the/ do not interfere)
Equi pment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 2 mm ID glass packed with Chromosorb 102 80/100 mesh
-------
3rd Update - August 1982 Epichlorohydrin EPA-1
Note: This packing material is not particularly suitable for
epichlorohydrin as a component in other formulations,
because of the strong absorptivity of this packing, and
the resultant necessity for baking-off the absorbed
materials at high temperatures. It is suggested that
formulations be extracted by column chroma tography.
3. Precision liquid syringe
k. Usual laboratory glassware
Operating conditions for FID;
Column temperature: 1 50°C
Injection port temperature: 200°C
Detector temperature: 200°C
Carrier gas (helium or nitrogen), flow adjusted as necessary
Hydrogen flow: adjusted as necessary
Air flow: adjusted as necessary
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi 1ity.
Procedu re;
Preparation of standard;
Weigh 150 mg epichlorohydrin standard into a 100 ml volumetric flask, make to
volume with acetone, and mix well, (cone 1.5 mg/ml)
Preparation of sample;
Weigh a portion of sample equivalent to 150 mg epichlorohydrin into a 100 ml
volumetric flask, make to volume with acetone, and mix well, (cone 1.5 mg/ml)
GC Determination;
Inject 2 to 3 ul standard and, if necessary, adjust instrument parameters and
the volume injects to give peak heights of 1/2 to 3/^ full scale. Proceed
with the determination, making at least three injections each of standard and
sample solutions.
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3rd Update - August 1982 Epichlorohydrin EPA-1
Calculation;
Measure the peak heights or areas of both standard and sample and calculate
the percent epichlorohydrin as follows:
(peak height or area sample)(weight standard injected)($ purity)
(peak height or area standard)(weight sample injected)
Method submitted by EPA, CBIB, Beltsville Chemistry Lab, Beltsville, MD
(Elmer H. Hayes and Mark W. Law)
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982 Fonofos EPA-1
Determination of Fonofos by Infrared Spectroscopy
Fonofos is the accepted (BSI, ISO) common name for 0-ethyl S-phenyl
ethylphosphonodithioate, a registered insecticide having the chemical
structure:
Molecular formula: CinH.,
lu \? f.
Molecular weight: 246.3
Physical state-color-odor: light yellow liquid with a pungent, mercaptan-like
odor
Boiling point: 130°C at 0.1 mm Hg
Solubility: practically insoluble in water; miscible with organic solvents
such as: acetone, kerosene, methyl isobutyl ketone, xylene
Stability: stable under normal conditions
Other names: Dyfonate; N-2790
Reagent s_;
1. Fonofos standard of known purity
2. Carbon disulfide, pesticide or spectro grade
3- Sodium sulfate, anhydrous, granular
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3rd Update - August 1982 Fonofos EPA-1
Equipment;
1. Infrared spectrophotometer, double beam, with matched 0.2 mm KBr cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
k. Usual laboratory glassware
Procedure:
Preparation of standard;
Weigh 100 mg fonofos standard into a 10 ml volumetric flask, dissolve in and
make to volume with carbon disulfide. Add a little anhydrous sodium sulfate
to insure dryness. (cone 10 mg/ml)
Preparation of sample;
For liquids and emulsifiable concentrates - weigh a portion of sample
equivalent to 500 mg fonofos into a 50 ml volumetric flask, mix with and make
to volume with carbon disulfide. Add a little anhydrous sodium sulfate to
insure dryness.
For dusts, granules, and wettable powders - weigh a portion of sample
equivalent to 1 gram (1000 mg) fonofos into a 250 ml glass-stoppered flask or
screw-cap bottle, add 100 ml carbon disulfide by pipette, close tightly, and
shake on a mechanical shaker for one hour. Allow to settle, centrifuge or
filter if necessary, taking precautions to avoid evaporation of solvent. If
sample is of low percentage, it may be necessary to use a soxhlet extration
apparatus.
(cone 10 mg/ml)
IR Determination;
With carbon disulfide in the reference cell, and using the optimum
quantitative analytical settings for the particular IR instrument being used,
scan the standard and sample solutions from 690 to 5^0 cm (14.5
to 18.5 urn). Determine the absorbance of standard and sample using the peak
at 610 cm"' (16.^t urn) and a basepoint at 580 cm"1 917.25 urn).
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3rd Update - August 1982 Fonofos EPA-1
Calculat ion;
From the above absorbances and using the standard and sample concentrations,
calculate the percent fonofos as follows:
(abs. sample) (cone, std. in mg/ml) (% purity)
(abs. std.) (cone, sample in mg/ml)
This method was written in the general IR format used in this manual - it was
submitted by the State of Virginia - date unknown - in outline form. This
method has been used in the Beltsville Lab a few times, but has never been
checked thoroughly.
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Hexachlorophene EPA-1
Determination of Hexachlorophene by High Performance Liquid Chromatography
Hexachlorophene is the common name for 2,2-methylenebis (3,^,6-
trichlorophenol), a registered foliage fungicide and bactericide, plant
bactericide, and soil fungicide with some acaricidal activity. It has the
chemical structure:
OH
Cl Cl
Molecular formula:
Molecular weight: A06.9
Physical state-color-odor: white powder
Melting point: 16A to 165°C
Solubility: practically insoluble in water; soluble in acetone, alcohol,
chloroform, ether, propylene glycol, polyethylene glycol, olive
oil, cottonseed oil, and dilute aqueous solutions of the alkalis,
Stability: stable
Other names: Hexide; Nabac; Isobac (sodium salt)
Note: see end of method for a modified procedure using a RadialPak column and
methanol instead of acetonitrile.
-------
3rd Update - August 1982 Hexachlorophene EPA-1
Reagents:
1. Hexachlorophene standard of known purity
2. Acetonitrile/PIC A - (1 bottle PIC A in one liter of 90% acetonitrile +
10% water filtered through a 0.45 micron filter)
3. Water/PIC A - (1 bottle PIC A in one liter water filtered through a 0.^
micron fiIter)
k . Isopropanol, HPLC grade
Equipment;
1. High Performance Liquid Chromatograph with a variable wavelength UV
detector at 296 nm. If a variable wavelength detector is not available,
operating parameters and concentrations may have to be changed to obtain
the necessary separation and sensitivity.
2. Column: uBondapak C18 (30 cm x 3-9 mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.^5 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 8?% (90% acetonitrile/10% water/PIC A) + 23% (water/PIC A)
Column temperature: ambient
Flow rate: 1.5 ml/min
Wavelength: 296 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg hexachlorophene standard into a 100 ml volumetric flask, dissolve
in and make to volume with isopropanol; mix well. Filter a portion through a
0.^5 micron filter, (cone 1 mg/ml)
Preparation of sample;
Weigh a portion of sample (liquid or E. C.) equivalent to 100 mg hexachloro-
phene into a 100 ml volumetric flask, mix with and make to volume with
isopropanol; mix thoroughly. Filter a portion through a 0.^5 micron filter.
(cone 1 mg/ml) For dry formulations, use a 125 ml screw-cap flask and add
100 ml isopropanol by pipette. Shake for 30 minutes, and filter a portion.
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3rd Update - August 1982 Hexachlorophene EPA-1
HPLC Determination;
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample solutions.
Calculat ion;
Measure the peak height or peak area for each peak and calculate the average
for both standard and sample. Using these averages, calculate the percent
hexachlorophene as follows:
(peak height or area sample)(weight standard injected)(% purity standard)
(peak height or area standard) (weight sample injected)
Method submitted by Mark W. Law, EPA, Beltsville, MD March 19?8
The following modification of the above method was developed at the several
HPLC schools sponsored by EPA over the last few years.
Column: Radial Pak C18
Mobile phase: 90? (30% methanol/10? water/PIC A) + 10? (water/PIC A)
Flow rate: 8 ml/min
Amount injected: 20 ul
All other parameters, concentrations, and calculations are the same as given
above.
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
-------
3rd Update - August 1982
Lamprecid (Trade Name) EPA-1
Determination of Lamprecid by Ultraviolet Spectroscopy
Lamprecid is the trade name (Hoechst AG - West Germany) for alpha, alpha,
alpha-trifluoro-4-nitro-meta-cresol. Lamprecid is a selective fish killer
used to control sea lampreys. Its chemical structure is:
NO,
Molecular formula: CyhYF,NO, (free phenol)
Molecular weight: 207.11
229.09
(sodium salt)
Physical state-color-odor: solid
Melting point: 76°C
Solubility: soluble in ethanol; sodium salt is very water soluble
Stabi1ity:
Other names: Dowlap: TFM; 3-trifluoro-^-nitrophenol; A-nitro-3-
(trifluoromethyl) phenol
Reagents;
1. Lamprecid standard of known purity
2. Sodium hydroxide, 1N aqueous solution
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3rd Update - August 1982 Lamprecid (Trade Name) EPA-1
Equipment!
1. Ultraviolet spectrophotometer, double beam ratio recording
with matched 1 cm cells
2. Usual laboratory volumetric glassware
Procedure:
Preparation of standard;
Weigh an amount of standard Lamprecid equivalent to 100 mg of 100& purity into
a 100 ml volumetric flask, add 10 ml 1 N NaOH solution, and make to volume
with water. Mix thoroughly and pipette 5 ml into a 1000 ml volumetric flask
and make to volume with water, (final cone 5 ug/ml)
Preparation of Sample;
Weigh an amount of sample equivalent to 100 mg Lamprecid into a 100 ml
volumetric flask, add 10 ml 1 N NaOH solution, and make to volume with
water. Mix thoroughly; pipette 10 ml into a 1000 ml volumetric flask
and make to volume with water, (final cone 5 ug Lamprecid/ml)
UV Determination;
With the spectrophotometer at the optimum quantitative settings for the
particular instrument being used, balance the pen at 0 and 100% transmission
at 395 nm with a blank reagent solution* in each cell.
*blank reagent solution - 10 ml IN NaOH solution diluted to 100 ml, then 5 ml
diluted to 1000 ml.
Scan both standard and sample solutions from 500 to 200 nm with blank reagent
solution in the reference cell. Measure the absorbance of standard and sample
solutions at 395 nm using a baseline from 310 to 280 nm.
Calculat ions;
From the above absorbances and using the standard and sample concentrations,
calculate the percent Lamprecid as follows;
(abs. sample) (cone. std. in ug/ml) (% purity)
"Tabs, std.)(cone, sample in ug/ml)
This method has been used successfully in the Beltsville Chemistry Laboratory
several times in the past - the late 1960's and early 1970's; however no new
data is available. Any information about analysis of Lamprecid will be
apprec iated.
-------
3rd Update - August 1982 Malathion EPA-3
Determination of Malathion by High Performance Liquid Chromatography
Malathion is the accepted (ANSI, BSI, ISO) common name for Qf 0-dimethyl
dithiophosphate of diethyl mercaptosuccinate, a registered insecticide having
the chemical structure:
CH, 0. jj ?<
">P_ — s CH C 0 CH2 CH3
CH, CT
3
0
II
CH2 — C C
) CH2— - CH3
Molecular formula: ClgH1_OgPS2
Molecular weight: 330.4
Physical state-color-odor: clear, colorless to amber liquid; technical grade
35% has a garlic-like odor
Melting point: 2.85°C
Boiling point: 156 to 157°C at 0.7 mm Hg with slight decomposition
Solubi1i ty: 1^5 ppm in water; 1imited solubi1i ty in petroleum oils but
miscible with most organic solvents; light petroleum oil (30-
60°C) is soluble in malathion to the extent of 35%
Stability: rapidly hydrolyzed at pH above 7-0 or below 5-0 but is stable in
aqueous solutions buffered at pH 5-26; incompatible with alkaline
pesticides and Is corrosive to iron, hence lined containers must
be used
Other names: mercaptothion (So. Africa); carbofos (USSR); mercaptotion
(Argentina); maldison (Australia); Calmathion; Celthion;
Cythion; Detmol MA 36% (Albert & Co. Germany); Emmatos; Emmatos
Extra; ForMal; Fyfanon; Hi 1thion; Karbofos; Kop-Thion; Kypfos;
-------
3rd Update - August 1982 Malathion EPA-3
Malaspray; Malamar; Malaphele; Malathion; Malathion ULV
Concentrate; Malatol; Maimed; Maltox; MLT; Sumitox; Vegfru
Malatox; Zithol 0,0-dimethyl S-(1,2-dicarbethoxyethyl)
phosphorodithioate; S-[1,2-di(ethoxycarbonyl)-ethyl] dimethyl
phosphoroth iolothionate
Reagents;
1. Malathion standard of known purity
2. Benzyl benzoate (internal standard) of known purity
3. Acetonitrile, HPLC grade
*». Methanol, HPLC grade
5. Internal standard solution - weigh 100 mg benzyl benzoate into a 100 ml
volumetric flask, dissolve in and make to volume with methanol; mix
thoroughly. Dilute 50 ml to 500 ml and mix well, (cone 0.1 mg/ml)
Equipment;
1. High Performance Liquid Chromatograph with UV detector at 251* nm. If a
variable wavelength detector is available, other wavelengths may be used
to increase sensitivity or to eliminate interference.
2. Column: uBondapak C18 (30 cm x 3.9 mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 60% acetonitrile + kQ% water
Column temperature: 33°C
Flow rate: 2 ml/min
Wavelength: 25^ nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg malathion standard into a 125 ml screw-cap flask, add 100 ml
internal standard solution by pipette, close tightly, and shake to dissolve,
Filter a portion through a 0.^5 micron filter, (cone 1 mg malathion and
0.1 mg benzyl benzoate per ml)
-------
3rd Update - August 1982 Malathion EPA-3
Preparation of sample:
Weigh a portion of sample equivalent to 100 mg malathion into a 125 nil screw-
cap flask, add 100 ml internal standard solution by pipette, close tightly and
place in an ultrasonic bath for several minutes, then shake on a mechanical
shaker for one hour. Filter through a 0.^5 micron filter, (cone as above)
HPLC Determination;
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample.
Calculat ion;
Measure the peak heights or areas of the malathion and the benzyl benzoate for
both the standard and sample solutions and calculate the following ratios:
peak height or area malathion
Ratio of standard = —, . . . —
peak height or area benzyl benzoate
peak height or area malathion
Ratio of sample =
peak height or area benzyl benzoate
Average the standard and sample ratios, and calculate the percent malathion as
follows:
(ratio of sample) (weight standard) (% purity of standard)
(ratio of standard) (weight sample)
Method submitted by EPA - NEIC, Denver, Colorado (G. Thomas Gale)
January 1980
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982 Methazole EPA-1
Determination of Methazole by Infrared Spectroscopy
Methazole is the accepted (ANSI, BSI, WSSA) common name for 2-(3,lt-
dichlorophenyl)-4-methyl-1,2,4-oxadiazol idine-3,5-dione, a registered
herbicide havinq the chemical structure:
0
Molecular formula: CgHgC^I^O,
Molecular weight: 261.1
Physical state-color-odor: tan, dry solid; odorless when pure
Melting point: 123 to 124°C
Solubility: 1.5 ppm in water at 25°C; 0.65% in methanol; 5.5% in xylene;
9.0% in acetone; soluble in chloroform and benzene
Stability: decomposes before boiling; subject to some decomposition by
germicidal UV when dissolved in methanol and to sunlight when
dissolved in water.
Other names: oxydiazol; Probe: VCS 438
Reagents;
1. Methazole standard of known purity
2. Chloroform, pesticide or spectro grade
3. Sodium sulfate, anhydrous, granular
-------
3rd Update - August 1982 Methazole EPA-1
Equipment;
1. Infrared spectrophotometer, double beam with matched Q.5mm NaC1 cells
2. Mechanical shaker
3. Centrifuge or filtration apparatus
*f. Usual laboratory glassware
Procedure:
Preparation of standard;
Weigh 100 mg methazole standard into a small glass-stoppered flask or screw-
cap bottle, add 50 ml chloroform by pipette and a little anhydrous sodium
sulfate to insure dryness, shake thoroughly, and allow to settle.
(cone 2 mg/ml)
Preparation of sample;
Weigh a portion of sample equivalent to 200 mg methazole into a 250 ml glass-
stoppered flask or screw-cap bottle, add 100 ml chloroform by pipette and some
anhydrous sodium sulfate. Shake on a mechanical shaker for about one hour.
Allow to settle, centrifuge or filter if necessary taking precautions to avoid
evaporation of solvent, (cone 2 mg/ml)
IR Determinat ion:
With chloroform in the reference cell, and using the optimum quantitative
analytical settings for the particular IR instrument being used, scan both
standard and sample solutions from 1538 to 1818 cm (6.5 to 5-5 urn). with a
horizontal baseline from 1960 to 1666 cm" (5.1 to 6.0 urn).
Calculat ion:
From the above absorbances and using the standard and sample concentrations,
calculate the percent methazole as follows:
(abs. sample) (cone. std. in mg/ml) (% purity)
(abs. std.) (cone, sample in mg/ml)
The absorbance is linear from 0.8 to 3»2 mg/ml.
-------
3rd Update - August 1982 Methazole EPA-1
Method submitted by EPA (former) Product Analysis Laboratory, Region II, New
York, NY
March 1977
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
-------
3rd Update - August 1982 Methidathion EPA-2
Determination of Methidathion by Gas Chromatography (FID-IS)
Methidathion is the accepted (ANSI, BSI, ISO) common name for 0,0-
dimethyl phosphorodithioate S-ester with ^-(mercaptomethyl) 2-methoxy-delta
2-1,3,^-thiadiazolin-5-one, a registered insecticide and acaricide having the
chemical structure:
CH3— OS - — 0_CH3
ij • -i — ~~" \_f fi _ " ..... • j.N •*
Molecular formula: CgH11
Molecular weight: 302.3
Physical state-color-odor: colorless crystals, characteristic odor of
organophosphates
Melting point: 39 to ^0°C
Solubility: 2^0 ppm in water at 25°C; readily soluble in acetone, benzene,
methanol
Stability: stable in neutral and weakly acid media but much less stable in
alkali; compatible with captan, thiram, zineb, and acaricides;
rapidly metabolized in plants
Other names: GS-13005; Supracide; Ultracide; S-(2,3-dihydro-5-methoxy-2-oxo-
1,3,b-th?adiazol-3-ylmethyl) dimethyl phosphorothiolothionate;
S-[(5-methoxy-2-oxo-1,3,A-thiadiazol-3(2H-yl) methyl]0,0-
dimethyl phosphorodithioate
Reagents;
1. Methidathion standard of known purity
2. Dibutyl phthalate (internal standard), analytical grade
3. Acetone, pesticide grade
-------
3rd Update - August 1982 Methidathion EPA-2
k. Internal standard solution - weigh 100 mg dibutyl phthalate into a 100 ml
volumetric flask, dissolve in and make to volume with acetone, and mix
wel1. (cone 1 mg/ml)
Equipment:
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 1/4" glass packed with 3% OV-1 on 100/200 mesh Supelcoport
(or equivalent column)
3. Precision liquid syringe
A. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions for FID;
Column temperature: 190°C
Injection port temperature: 250°C
Detector temperature: 250°C
Carrier gas: nitrogen - 30 ml/min (adjusted as necessary)
Hydrogen flow: adjusted as necessary
Air flow: adjusted as necessary
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg methidathion standard into a small glass-stoppered flask or
polyseal-cap glass vial, add 25 ml internal standard solution by pipette, and
shake to dissolve, (cone k mg methidathion and 1 mg dibutyl phthalate per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 100 mg methidathion into a small flask
or vial as above, add 25 nil internal standard solution as above, and shake
thoroughly to dissolve and extract the methidathion. For course or granular
materials, shake mechanically for 10 to 15 minutes. Allow to settle and if
necessary centrifuge (or filter) to clarify, (cone k mg methidathion and 1 mg
dibutyl phthalate per ml)
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3rd Update - August 1982 Methidathion EPA-2
GC Determination;
Inject 2 ul of standard and, if necessary, adjust the instrument parameters
and the volume injected to give a complete separation within a reasonable time
and to obtain peak heights of 1/2 to 3A full scale. Proceed with the
determination, making at least three injections each of standard and sample
solutions. The elution order is dibutyl phthalate then methidathion.
Calculat ion:
Measure the peak heights or areas of the methidathion and dibutyl phthalate
for both the standard and sample solutions and calculate the following ratios:
peak height or area methidathion
Ratio of standard
Ratio of sample =
peak height or area dibutyl phthalate
peak height or area methidathion
peak height or area dibutyl phthalate
Average the standard and sample ratios, and calculate the percent methidathion
as follows:
(ratio of sample)(weight of standard)(& purity of standard)
(ratio of standard)(weight of sample)
Method submitted by NEIC, Denver, Colorado (Chuck Rzeszutko), August 1979
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982
Methoprene EPA-1
Determination of Methoprene by Gas Chromatography (FID-IS)
Methoprene is the accepted (ANSI) common name for isopropyl (2E,4E)-1V
methoxy-3,7,11-trimethyl-2,4-dodecadienoate, a registered insect growth
regulator having the chemical structure:
OCH,
Molecular formula: C.qH,.0,
Molecular weight: 310.5
Physical state-color-odor: amber liquid
Boiling point: 100°C at 0.05 mm Hg
Solubility: approximately 1.4 ppm in water; soluble in non-aqueous organic
solvents
Stabi1ity:
Other names: Altosid; Altosid Briquets; ZR-515
Reagents;
1. Methoprene standard of known purity
I. Dibutyl phthalate internal standard of known purity
3. Chloroform, pesticide grade
4. Internal standard solution - weigh 1.4 gram dibutyl phthalate into a
100 ml volumetric flask, dissolve in and make to volume with chloroform,
and mix well, (cone 14 mg/ml)
-------
3rd Update - August 1982 Methoprene EPA-1
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: V x 1/V glass packed with 3.8S SE-30 on 80 to 100 mesh
Diatoport S (or equivalent column)
3. Precision liquid syringe
k. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions for FiD;
Column temperature: 210°C
Injection port temperature: 225°C
Detector temperature: 230°C
Carrier gas: Helium - 20 ml/min (adjusted as necessary)
Hydrogen flow: 25 ml/min (adjusted as necessary)
Air flow: kQ ml/min (adjusted as necessary)
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi15ty.
Procedure:
Preparation of standard;
Weigh 110 mg methoprene standard into a 50 ml volumetric flask, add 5 ml
internal standard solution by pipette, make to volume with chloroform, and mix
thoroughly. Pipette a 5 ml aliquot into a second 50 ml volumetric flask and
make to volume with chloroform; mix thoroughly, (cone 0.22 mg methoprene and
O.l^t mg dibutyl phthalate per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 110 mg methoprene into a 125 ml glass-
stoppered flask, add 5 ml internal standard solution by pipette, add k$ ml
chloroform (graduated cylinder or pipette), stopper tightly, and shake for one
hour on a mechanical shaker. Allow to settle, centrifuge or filter a portion
if necessary, taking precaution to avoid evaporation of chloroform.
Dilute a 5 ml portion to 50 ml as above, (cone as above)
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3rd Update - August 1982 Methoprene EPA-1
GC Determination;
Inject 1 to 2 ul of standard and, if necessary, adjust the instrument
parameters and the volume injected to give a complete separation within a
reasonable time and to obtain peak heights of 1/2 to 3A full scale. Proceed
with the determination making at least three injections each of standard and
sample solutions. The elution order is dibutyl phtnalate then methoprene.
Calculation;
Measure the peak heights or areas of the methoprene and dibutyl phthalate for
both the standard and sample solutions and calculate the following ratios:
peak height or area methoprene
Ratio standard =
Ratio sample
peak height or area dibutyl phthalate
peak height or area methoprene
peak height or area dibutyl phthalate
Average the standard and sample ratios, and calculate the percent methoprene
as follows:
(ratio sample) (weight standard) (% purity standard)
.1-- •
(ratio standard) (weight sample)
Method submitted by EPA (former) Product Analysis Laboratory, Region II,
New York, NY
January 1977
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982 Methyl Nonyl Ketone EPA-1
Determination of Methyl Nonyl Ketone by Gas Chrcmatography (TCD-IS)
Methyl nonyl ketone is the popular name for 2-undecanone, a registered
animal repellent having the chemical structure:
0
II
-— CH~CH-^CH—CH-^CH~CH— CH— CH— C— CH
Molecular formula: C^H-.jO
Molecular weight: 170.3
Physical state-color-odor: clear liquid
Melting point: 11 to 13°C
Boiling point: 231.5 to 232. 5°C (technical 35% purity - 223°C)
Solubility: insoluble in water; miscible with petroleum distillates and most
other common organic solvents
Stability: effectiveness as a repellent last about 24 hours
Other names: MGK Dog and Cat Repellent
Reagents :
1. Methyl nonyl ketone standard of known purity
2. 2-ethyl -1 ,3-hexanediol (internal standard), analytical grade
3. Acetone, pesticide grade
4. Internal standard solution - weigh 2.5 grams 2-ethyl -1 ,3-hexanediol into a
100 ml volumetric flask, make to volume with acetone, and mix well .
(cone 25 mg/ml )
-------
3rd Update - August 1982 Methyl Nonyl Ketone EPA-1
Eggipment;
1. Gas chromatograph with a thermal conductivity detector (TCD)
2. Column: 6' x 1/8" SS packed with 10% SE-30 on 80/100 Diatoport S or
V x 1/V glass packed with 3.8£ SE-30 on 80/100 Diatoport S
(or equivalent column)
3. Precision liquid syringe
4. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions for TCP:
Column temperature: 145°C for 1/8" column; 120°C for 1/4" column
Injection port temperature: 225°C
Detector temperature: 150°C
Filament current: 200 ma
Carrier gas: helium - flow adjusted as necessary
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi !ity.
Procedure:
Preparation of standard;
Weigh 400 mg methyl nonyl ketone standard into a 25 ml volumetric flask, add
10 ml internal standard solution by pipette, and make to volume with acetone;
mix well, (cone 16 mg methyl nonyl ketone and 10 mg internal standard per ml)
Preparation of sample;
For liquid samples, weigh an amount equivalent to 400 mg methyl nonyl ketone
into a 25 ml volumetric flask, add 10 ml internal standard solution by
pipette, and make to volume with acetone; mix well.
For granules and dusts, weigh a portion of sample equivalent to 400 mg methyl
nonyl ketone into a small glass-stoppered flask or screw-cap bottle, add 10 ml
of internal standard solution by pipette and 15 ml acetone by pipette, close
tightly and shake on a mechanical shaker for 30 minutes. Allow to settle, and
if necessary centrifuge or filter to clarify.
(cone 16 mg methyl nonyl ketone and 10 mg internal standard per ml)
-------
3rd Update - August 1982 Methyl Nonyl Ketone EPA-1
GC Determination;
Inject 3 to 5 ul of standard and, if necessary, adjust the instrument
parameters and the volume injected to give a complete separation within a
reasonable time and to obtain peak heights of 1/2 to 3/4 full scale. Proceed
with the determination, making at least three injections each of standard and
sample solutions. The elution order is 2-ethyl-1,3-hexanediol then methyl
nonyl ketone.
Calculations;
Measure the peak heights or areas of the methyl nonyl ketone and the 2-ethyl~
1,3-hexanediol for both standard and sample solutions and calculate the
rat ios;
peak height or area methyl nonyl ketone
Ratio of standard = . ' ' ——r
peak height or area 2-ethyl-1,3-hexanedlol
peak height or area methyl nonyl ketone
Ratio of sample =
peak height or area 2-ethyl-1,3-hexanediol
Average the standard and sample ratios, and calculate the percent methyl nonyl
ketone as follows:
(ratio of sample)(weIght of standard)(^ purity of standard)
(ratio of standard) (weight of sample)
Method submitted by EPA (former) Product Analysis Laboratory, Region II,
New York, NY January 1976
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982 Methyl Parathion EPA-6
Determination of Methyl Parathion by High Performance Liquid Chromatography
Methyl parathion is the accepted (BSI, ISO) common name for 0,0-dimethyl-
C-p-nitrophenyl phosphorothioate, a registered insecticide having the chemical
structure:
Molecular formula: CgH^NQ^PS
Molecular weight: 263-2
Physical state-color-odor: white crystalline solid; the technical product is
a light to dark tan liquid of about 80% purity,
crystallizing at about 29°C
Melting point: 35 to 36°C
Solubility: 55 to 60 ppm in water at 25°C; slightly soluble in light mineral
and petroleum oils; soluble in most other organic solvents
Stability: hydrolyzed by alkalis; compatible with most other pesticides
except alkaline materials; isomerizes on heating; it is a good
methylating agent
Other names: Cekumethion; Devithion; Dimethyl Parathion; Drexel Methyl
Parathion AE; E601 ; Folidol M; Fosferno M50; Gearphos; Metacide;
Metaphos; Metron; Nitrox 80; Parataf; Paratox; Partron M;
Penncap-M; Tekwaisa; Vertac Methyl Parathion Technisch 80%;
Wofatox
Reagents;
1. Methyl parathion standard of known purity
2. Methanol, HPLC grade
3. Water, HPLC grade
A. Acetic acid, ACS
-------
3rd Update - August 1982 Methyl Parathion EPA-6
Equ ipment;
1. High Performance Liquid Chromatograph with UV detector at 254 nm. If a
variable wavelength detector is available, other wavelengths may be used
to increase sensitivity or to eliminate interference.
2. Column: Radial-Pak C18 or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions:
Mobile phase: 11% methanol + 22% water + U acetic acid (one solution)
Column temperature: ambient
Flow rate: 5 to 7 ml/min
Wavelength: 254 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg methyl parathion into a 125 ml screw-cap flask, add 100 ml
methanol by pipette, close tightly, and shake to dissolve. Filter through a
0.45 micron filter, (cone 1 mg/ml)
Preparation of sample;
Weigh a portion of sample equivalent to 100 ml methyl parathion into a 125 ml
screw-cap flask, add 100 ml methanol by pipette, close tightly, and shake for
30 minutes on a mechanical shaker. (A few minutes in an ultrasonic bath may
help to effect solution) Filter through a 0.45 micron filter, (cone 1 mg/ml)
HPLC Determination;
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample solutions.
-------
3rd Update - August 1982 Methyl Parathion EPA-6
Calculation;
Measure the peak height or peak area for each peak and calculate the average
for both standard and sample. Using these averages, calculate the percent
methyl parathion as follows:
(peak height or area sample)(weight standard injected)(£ purity standard)
(peak height or area standard)(weight sample Injected)
This is a modification of Methyl Parathion EPA-1 for using a Radial-Pak column
and acid suppression.
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982
Mixed Pesticides EPA-2
(Atrazine 6 Metolachlor)
Determination of Atrazine and Metolachlor Mixtures
by Gas Chroma tography (FID-IS)
Atrazine:
Atrazine is the accepted (ANSI, BSI, ISO, WSSA) common name for
2-ch1oro-A-ethylamino-6-isopropylamfno-1,3,5-triazlne, a registered herbicide
having the chemical structure:
Cl
CH,
,CH
H
• N-
H
,C — N CH
CH,
CH,
Molecular formula:
Molecular weight: 215.7
Physical state-color-odor: colorless crystalline solid
Melting point: 173 to 175°C
Solubility: 33 ppm in water at 25°C; 1.2$ in ethyl ether; 1.8? in methanol;
2.8% in ethyl acetate? 5.2% in chlorpform; 18.3$ in dimethyl
suIfoxide
Stability: stable in neutral and slightly acidic or basic media; hydrolyzes
In acid and alkaline conditions of higher temperatures to the
herbicidally inactive hydroxy derivative; non-flammable; non-
corrosive under normal use conditions; very stable shelf life with
only slight sensitivity to natural light and extreme temperature;
compatible with most other pesticides
Other names:
AAtrex; Atranex; Atratol; Crisatrina; Crisazine; G 30027;
Gesaprim; Griffex; Primatol A; Shell Atrazine Herbicide; Vectal
SC; 6-chloro-N-ethyl-N'-(1-methylethyl)-1,3,5-triazine-2,4-
diamine
-------
3rd Update - August 1982 Mixed Pesticides EPA-2
(Atrazine & Metolachlor)
Metolachlor;
Metolachlor is the accepted (ANSI, ISO, WSSA) common name for 2-
chloro-N-(2-ethyl-6-methylphenyl)-N-(2-methoxy-1-methylethyl) acetamide, a
registered herbicide having the chemical structure:
O J-f _ — — U rl o
Molecular formula: C.-^NOpd
Molecular weight: 283.8
Physical state-color-odor: odorless, white to tan liquid
Boiling point: 100°C at 0.001 mm Hg
Solubility: 530 ppm in water at: 20°C; miscible with xylene, toluene, dimethyl
formamide, methyl cellusolve, butyl cellusolve, ethylene
dichloride, and cyclohexanone; insoluble in ethylene glycol and
propylene glycol
Stability: compatible with most pesticides and fluid fertilizers when used at
normal rates; non-corrosive to steel or tin; not harmful to
plastic or fiberglass spray tanks; shelf life estimated to be 5
years minimum based on no significant decomposition at 70°C for 3
weeks or at 50°C for 20 weeks; no crystallization at temperatures
below 0°C
Other names: Bicep; CGA-2^705; Codal; Cotoran Multi; Milocep; Ontrack 8E;
Primagram; Primatex
-------
3rd Update - August 1982 Mixed Pesticides EPA-2
(Atrazine & Metol&chlor)
Reagents;
1. Atrazine standard of known purity
2. Metolachlor standard of known purity
3. Alachlor (internal standard) of known purity
A. Acetone, pesticide grade
5. Internal standard solution - weigh 500 mg alachlor into a 50 ml volumetric
flask, dissolve in and make to volume with acetone; mix well.
(cone 10 mg/ml)
Equipment;
1. Gas chromatograph with a flame ionization detector (FID)
2. Column: 6' x 2 mm ID glass packed with 3& SE-30 on 100/120 mesh
Chromosorb W HP
3. Precision liquid syringe: 10 ul
J*. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions;
Column temperature: 150°C
Injection port temperature: 250°C
Detector temperature: 250°C
Carrier gas: helium - flow: adjusted as necessary
Hydrogen flow: adjusted as necessary
Air flow: adjusted as necessary
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 80 mg each of atrazine standard and metolachlor standard into a 100 ml
volumetric flask, add 10 ml internal standard solution by pipette, make to
volume with acetone, close tightly and mix thoroughly, (cone 0.8 mg atrazine,
0.8 mg metolachlor, and 1 mg alachlor per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 80 mg atrazine and/or metolachlor (if
the percent of atrazine and metolachlor differ too much for one solution, make
two solutions) into a 100 ml volumetric flask, add 10 ml internal solution,
make to volume with acetone, close tightly and mix thoroughly.
-------
3rd Update - August 1982 Mixed Pesticides EPA-2
(Atrazine & Metolachlor)
(cone as above)
GC Determination;
Inject 1 or 2 ul of standard and,, if necessary, adjust the instrument
parameters and the volume injected to give a complete separation within a
reasonable time and to obtain peak heights of 1/2 to 3/^ full scale. Proceed
with the determination, making at least three injections each of standard and
sample solutions. The elution order is atrazine, alachlor, then metolachlor.
Calculation;
Measure the peak heights or areas of the atrazine, alachlor, and metolachlor
for both standard and sample solutions and calculate the following ratios;
peak height or area atrazine
atrazine: ratio of standard =—
ratio of sample
peak height or area alachlor
peak height or area atrazine
metolachlor; ratio of standard =
ratio of sample
peak height or area alachlor
peak height or area metolachlor
peak height or area alachlor
peak height or area metolachlor
peak height or area alachlor
Average the standard and sample ratios, and calculate the percent atrazine and
metolachlor as follows:
(ratio of sample)(weight standard)(% purity standard)
% atrazine = , : — . . . , —<
(ratio of standard) (weight sample)
(ratio of sample)(weight standard)(% purity standard)
metolachlor
(ratio of standard) (weight sample)
This method is an adaptation of Atrazine EPA-2 (Tentative) October 1975
EPA Beltsville Chemistry Lab, Beltsville, MD (Jack B. Looker)
April 1978
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982
Mixed Pesticides EPA-3
(Disulfoton & Fensulfothion)
Determination of Disulfoton and Fensulfothion Mixtures
by Gas Chromatography (FID-IS)
Disulfoton:
Disulfoton is the accepted (BSI, ISO) common name for 0,0-
dimethyl S- [2-(ethyl th io)ethy 1 jphosphorodi thioate, a registered insecticide
and acaricide having the chemical structure:
CH— -CH
v
\!f
r
^
Molecular formula:
Molecular weight:
Physical state-color-odor: pure - colorless to pale yellow liquid with a
characteristic odor of sulfur compounds
technical - dark yellowish oil with an aromatic
odor
Boiling point: 62°C at 0.1 mm Hg
Solubility: 25 ppm in water at room temperature; readily soluble in most
organic solvents
Stability: subject to hydrolysis under alkaline conditions; stable in normal
storage
Other names: Bay 276; Bay 19&39; Disyston; Di-Syston (in U.S.);
di thiodemeton; di thiosustox; Frumin AL; M-7*» (USSR); Solvirex
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3rd Update - August 1982
Mixed Pesticides EPA-3
(Disulfoton & Fensulfothion)
Fensulfothion;
Fensulfothion is the accepted (BSI, ISO) common name for 0,0-
diathyl 0-[l»-(methyIsulfinyl)pheny1]phosphorothioate, a registered insecticide
and nematicide having the chemical structure:
•CH2 0
.0
— o—(/ \S~s—
CH,
Molecular formula: C. .H,.,0. PS-
Molecular weight: 308.35
Physical state-color-odor: oily yellowish-brown liquid
Boiling point: 138 to
at 0.0 mm Hg
Solubility: slightly soluble in water (1600 ppm); soluble in most organic
solvents except aliphatic
Stability: believed to be compatible with most insecticides and fungicides
except alkaline materials; subject to hydrolysis; readily oxidized
to the sulphone; isomerizes readily to the S ethyl isomer
Other names: Bay 251^*1; Dasanit; S7&7; Terracur P; diethyl-p-
methyIsulfinylphenyl thiophosphate
Reagents;
1. Disulfoton standard of known purity
2. Fensulfothion standard of known purity
3. Dipentyl phthalate (internal standard) of known purity
k. Acetone, pesticide grade
5. Internal standard solution - weigh 750 mg dipentyl phthalate into a 50 ml
volumetric flask, dissolve in and make to volume with acetone; mix well.
(cone 15 mg/ml)
-------
3rd Update - August 1982 Mixed Pesticides EPA-3
(Disu]foton & Fensulfothion)
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 2 mm ID glass packed with 5% SE-30 on 80/100 mesh Chromosorb
W HP (or equivalent column)
3. Precision liquid syringe: 10 ul
k. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions for FID;
Column temperature: 210°C
Injection port temperature: 250°C
Detector temperature: 250°C
Carrier gas: helium or nitrogen - flow: adjusted as necessary
Hydrogen flow: adjusted as necessary
Air flow: adjusted as necessary
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg disulfoton and 100 mg fensulfothion standards into a 50 ml
volumetric flask, add 5 ml internal standard solution by pipette, make to
volume with acetone, close tightly and mix thoroughly, (cone 2 mg disulfoton,
2 mg fensulfothion, and 1.5 mg dipentyl phthalate per ml)
Preparation of sample:
Weigh a portion of sample equivalent to 100 mg disulfoton and/or fensulfothion
(if the percent of disulfoton and fensulfothion differ too much for one
solution, make two solutions) into a 50 ml volumetric flask, add 5 ml internal
standard solution by pipette, make to volume with acetone, close tightly and
mix thoroughly, (cone as above)
For solid samples, use a 125 ml screw-cap flask instead of a 100 ml volumetric
flask and add 5 ml internal standard solution and kS ml acetone by pipette)
GC Determination;
Inject 5 ul of standard and, if necessary adjust the instrument parameters and
the volume injected to give a complete separation within a reasonable time and
to obtain peak heights of 1/2 to 3A full scale. Proceed with the
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3rd Update - August 1982 Mixed Pesticides EPA-3
(Disulfoton 6 Fensulfothion)
determination, making at least three injections each of standard and sample
solutions. The elution order is disulfoton, dipentyl phthalate, then
fensulfothion.
Calculat ior^t
Measure the peak heights or areas of the disulfoton, dipentyl phthalate, and
fensulfothion for both standard and sample solutions and calculate the
following ratios:
,. ., , , . peak height or area disulfoton
disulfoton: ratio of standard =
rat io of sample
peak height or area dipentyl phthalate
peak height or area disulfoton
peak height or area dipentyl phthalate
, ., . . f peak height or area fensulfothion
fensulfothion: ratio of standard =
peak height or area dipentyl phthalate
peak height or area fensulfothion
rat 10 of sample =
peak height or area dipentyl phthalate
Average the standard and sample ratios, and calculate the percent disulfoton
and fensulfothion as follows:
(ratio of sample)(weight standard)(% purity standard)
(ratio of standard) (weight sample)
(ratio of sample)(weight standard)(% purity standard)
(ratio of standard) (weight sample)
% fensulfothion
Method submitted by Mark W. Law, EPA Chemistry Lab, Beltsville, MD
March 1976
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982
Naphthaleneacetic acid EPA-1
Determination of Naphthaleneacetic acid and Its Ammonium Salt
by High Performance Liquid Chroma tog raphy
Naphthaleneacetic acid is the accepted (BSI, ISO) common name for
1-naphthaleneacetic acid, a registered plant growth regulator having the
chemical structure:
Molecular formula: ^
Molecular weight: 186.21
Physical state-color-odor: odorless, white crystals or amorphorous powder
Melting point: 134 to 125°C
Solubility: ^20 ppm in water at 20°C; slightly soluble in carbon
tetrachloride and xylene; very soluble in acetone, chloroform,
ethanol, and isopropanol
Stability: non-flammable; non-corrosive; stable on storage; compatible with
other pesticides
Other names: Celmone; Fruitone N; NAA; NAA 800; Nafusaku; Phyomone; Planofix;
Plucker; Primacol; Rootone; Stik; Tekkam; TipOff; Transplantone;
Tre-Hold
Reagents;
1. Naphthaleneacetic acid standard of known purity
2. Methanol, ACS
-------
3rd Update - August 1982 Naphthaleneacetic acid EPA-1
3. 1% Acetic acid in methanol solution
4. 0.0025M Phosphoric acid aqueous solution
Equipment;
1. High Performance Liquid Chroma tograph with a variable wavelength UV
detector at 272 nm. If a variable wavelength detector is not available,
operating parameters and concentrations may have to be changed to obtain
the necessary separation and sensitivity.
2. Column: uBondapak C18 (30 cm x 3-9 mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 65% Methanol + 35% 0.0025M Phosphoric acid aqueous solution
Column temperature: 55°C (ambient temperature could be used with a change in
parameters)
Flow rate: 1.2 ml/min
Wavelength: 272 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg naphthaleneacetic acid standard into a 100 ml volumetric flask,
dissolve in and make to volume with 1% acetic acid - methanol solution. Mix
well and dilute a 25 ml aliquot to 100 ml with the acetic acid - methanol
solution. Filter a portion through a Q.kS micron filter, (cone 0.25 mg/ml)
Preparation of sample:
Weigh a portion of sample equivalent to 100 mg naphthaleneacetic acid into a
100 ml volumetric flask, dissolve in and make to volume with }% acetic acid -
methanol solution. Mix well and dilute 25 ml to 100 ml as above. Filter a
portion through a 0.^5 micron filter, (cone as above)
-------
3rd Update - August 1982 Naphthaleneacetic acid EPA-1
HPLC Determination!
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample solutions.
Calculat ion;
Measure the peak height or area for each peak and calculate the average for
both standard and sample. Using these averages, calculate the percent
naphthaleneacetic acid as follows:
(peak height or area sample)(weight standard injected)(% purity standard)
(peak height or area standard) (weight sample injected)
Method submitted by E. S. Greer, EPA (formerly) Product Analysis Laboratory,
Region IX, San Francisco, California (Mr. Greer is now at Beltsville, MD)
August 1977
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
-------
3rd Update - August 1982 Naptalam EPA-1
Determination of Naptalam by Ultraviolet Spectroscopy
Naptalam is the accepted (BSI, ISO, WSSA) common name for N-1-
naphthylphthalamic acid, a registered herbicide having the chemical structure:
Molecular formula: C-gH.,NO_
Molecular weight: 291.3
Physical state-color-odor: purple crystalline powder with an unpleasant odor
Melting point: 185°C
Solubility: 200 ppm in water, 5900 ppm in acetone, 2100 ppm in isopropanol;
slightly soluble in benzene and ethanol; insoluble in hexane and
xylene; alkali metal salts are readily soluble in water
Stability: Hydrolyzed in solutions of pH more than 9«5j unstable at elevated
temperatures, tending to form the imide; non-corrosive;
non- explosive
Other names: Alanap; ACP 322; NPA; 60.8: Dyanap
Reagents;
1. Naptalam standard of known purity
2. Hexane, pesticide or spectro grade
3. Glacial acetic acid
4. Sodium hydroxide, 0.25N
-------
3rd Update - August 1982 Naptalam EPA-1
Equ i pmen t;
1. Ultraviolet spectrophotometer, double beam ratio recording with matched
1 cm eel Is.
2. Filtration apparatus, medium porosity fritted glass crucibles, buchner
funnels
3. Usual laboratory glassware
Procedure:
Preparation of standard;
Weigh 100 mg naptalam standard into a 100 ml volumetric flask, dissolve in and
make to volume with 0.25N NaOH. Mix thoroughly and pipette 5 ml into a 250 ml
volumetric flask; make to volume with distilled water and mix thoroughly.
(final cone 20 ug/ml)
Preparation of sample;
Weigh a portion of sample equivalent to 100 mg naptalam into a 50 ml beaker,
add 1 ml glacial acetic acid and mix thoroughly. Add 10 ml hexane, swirl, and
let stand until the naptalam precipitates (ten minutes or so). Filter through
a medium porosity fritted glass crucible and wash the beaker and filtered
precipitate three times with small amounts of hexane. Change buchner flask
and wash the filtered precipitate through the frittered glass crucible by
dissolving the precipitate in 0.25N NaOH. Rinse the 50 ml beaker with
portions of the 0.25N NaOH also. Transfer the filtrate to a 200 ml volumetric
flask and make to volume with a 0.25N NaOH. Mix thoroughly and pipette 10 ml
into a 250 ml volumetric flask; make to volume with distilled water and mix
thoroughly.
UV Determinat ion;
With the UV spectrophotometer at the optimum quantitative settings for the
particular instrument being used, balance the pen at 0 and 100% transmission
at 282 nm with distilled water in each cell. Scan both standard and sample
solutions from 350 to 220 nmwith distilled water in the reference eel 1.
-------
3rd Update - August 1982 Naptalam EPA-1
Calculations;
From the above absorbances and using the standard and sample concentrations,
calculate the percent naptalam as follows:
(abs. sample) (cone. std. in ug/ml) (% purity)
7* 55 _ .. .. ._. .. .
(abs. std.)(cone, sample in ug/ml)
Gravimetric factor: naptalam acid x 1.0755 = naptalam sodium salt
Method originally from Mississippi State Chemical Laboratory, Mississippi
State, Mississippi 39762.
Method checked by Jack Looker, Beltsville Chemistry Laboratory, CBIB, BFSD,
OPTS, EPA. There is a straight line relationship between absorbance and
concentration for up to 50 ug/ml.
Any criticism, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982 Neburon EPA-2
Determination of Neburon by Ultraviolet Spectroscopy
Neburon is the accepted (BSI, ISO, WSSA) common name for 1-n-butyl-3~
(3,4-dichlorophenyl)-1-methyl urea, a registered herbicide having the chemical
structure:
—CH2—CH2
Molecular formula: ^12^16^2^2^
Molecular weight: 275.18
Physical state-color-odor: odorless, white crystalline solid
Melting Point: 102 to 103°C
Solubility: 4.8 ppm in water at 2i»°C; very low in common hydrocarbon solvents
Stability: stable toward oxidation and moisture under normal storage
cond i t ions
Other names: Granurex; Kloben; Neburex
Reagents;
1. Neburon standard of known purity
2. Methylene chloride, pesticide or spectro grade
-------
3rd Update - August 1982 Neburon EPA-2
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording with matched
1 cm eel Is.
2. Mechanical shaker
3. Filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of standard:
Weigh 100 mg neburon standard into a 100 ml volumetric flask, dissolve in and
make to volume with methylene chloride. Mix thoroughly and pipette 10 ml into
a second 100 ml volumetric flask. Make to volume with methylene chloride, mix
thoroughly, and pipette 5 ml into a third 100 ml volumetric flask. Make to
volume with methylene chloride and mix thoroughly, (final cone 5 ug/ml).
Preparation of sample;
Weigh a portion of sample equivalent to 100 ml neburon into a 250 ml glass-
stoppered flask or screw-cap bottle, add 100 ml methylene chloride by pipette,
stopper tightly, and shake on a mechanical shaker for at least fifteen
minutes. Allow to settle, centrifuge or filter if necessry taking precautions
to avoid evaporation of solvent. Dilute 10 ml to 100 ml and then 5 ml to
100 ml as under standard preparation, (final cone 5 ug neburon/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative settings for the
particular instrument being used, balance the pen at 0 and 100% transmission
at 252 nm with methylene chloride in each cell. Scan both the standard and
sample solutions from 350 to 200 nmwith methylene chloride in the reference
cell. Measure the absorbance of standard and sample at 252 nm.
Calculat ions;
From the above absorbances and using the standard and sample concentrations,
calculate the percent neburon as follows:
<£ _ (abs. sample) (cone, std. in ug/ml) (% purity)
(abs. std.) (cone, sample in ug/ml)
-------
3rd Update - August 1982 Neburon EPA-2
Method submitted (summer - ?) 1978 by;
Dr. Gabriele Tartar!
Agrochemical Department
Control Laboratory
CIBA-GEIGY S.P.A.
C.P. 88
1-21047 SARONNO (VA)
ITALY
Note: The amount of standard and sample and some dilution factors have been
changed to allow more significant figures in the calculations and to reduce
errors in weighings and making dilutions. The final concentrations are as in
the method as received.
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Nicotine EPA-1
Determination of Nicotine by High Performance Liquid Chromatography
Nicotine is the trivial name for 3-(1-methyl-2-pyrrolidyl) pyridine, a
registered insecticide having the chemical structure:
Molecular formula:
Molecular weight: 162.2
Physical state-color-odor: colorless liquid alkaloid
Boi1 ing point: 247°C
Solubility: miscible with water below 60°C (forms a hydrate); miscible with
ethanol and ether; readily soluble in most organic solvents
Stability: very hydroscopic; darkens slowly and becomes viscous on exposure
to air; forms mono and dibasic salts with many acids and metals
Other names: Black Leaf
Reagents;
1. Nicotine standard of known purity
2. Phenol (Internal standard) of known purity (make sure that the phenol
gives a clean chromatogram with no co-eluting peaks)
3. Aqueous mobile phase - (0.0025M 1-heptane sulfonic acid sodium salt and
0.04M tetramethylammonium chloride adjusted to
pH 3.0 with sulfuric acid) Filter through a 0.^5
micron fi Iter .
A. Organic mobile phase - (0.06M tetramethylammonium chloride in 200 ml water
adjusted to pH 3.0 with sulfuric acid plus 800 ml
acetonitrile) Filter through a 0.^5 micron filter.
-------
3rd Update - August 1982 Nicotine EPA-1
5. Internal standard solution - weigh 1.1 grams phenol into a 500 ml
volumetric flask, dissolve in and make to volume with aqueous mobile
phase, (cone 2.2 mg/ml )
Equipment:
1. High Performance Liquid Chromatograph with UV detector at 25** nm. If a
variable wavelength detector is available, other wavelengths may be used
to increase sensitivity or to eliminate interference.
2. Column: MicroPak MCH-10 (30 cm x k mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 85% aqueous mobile phase + ^5% organic mobile phase
Column temperature: ambient
Flow rate: 1.5 to 2.0 ml/min
Wavelength: 254 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 95 mg nicotine standard into a 50 ml volumetric flask, make to volume
with internal standard solution, and mix well. Dilute 10 ml to 50 ml, mix
well, and filter through a 0.45 micron filter, (cone 0.38 mg nicotine and
2.2 mg phenol per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 95 mg nicotine into a 50 ml volumetric
flask, dissolve in and make to volume with internal standard solution, and mix
well. Dilute 10 ml to 50 ml, mix well, and filter through a 0.45 micron
filter, (cone 0.38 mg nicotine and 2.2 mg phenol per ml)
-------
3rd Update - August 1982 Nicotine EPA-1
HPLC Determination;
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or the amount injected to give good convenient
size peaks. Proceed with the determination making alternately three
injections each of standard and sample solutions. Elution order is nicotine
then phenol.
Calculation;
Measure the peak heights or areas of the nicotine and the phenol for both the
standard and sample solutions and calculate the following ratios;
peak height or area nicotine
Ratio of standard =
peak height or area phenol
peak height or area nicotine
Ratio of sample
peak height or area phenol
Average the standard and sample ratios, and calculate the percent nicotine as
fol lows::
(ratio of sample) (weight standard) (% purity of standard)
(ratio of standard)(weight sample)
Method submitted by EPA - NEIC, Denver, Colorado (G. Thomas Gale)
January 1980
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Paraquat EPA-1
Determination of Paraquat by High Performance Liquid Chroma tography
Paraquat is the accepted (ANSI, BSI, ISO, WSSA) name for 1,1'-dimethyl-
k,k'-bipyridylium ion; usually present as the dichloride or dimethyl sulfate
salt. Paraquat is a registered herbicide and desiccant and has the chemical
(cation) structure:
CH0 N
•CH,
2+
Molecular formula: C12H1AN2 (Cat'on)> C12H1^N2C12 (d'chl°r'de salt)
Molecular weight: 186.3 (cation); 257.2 (dichloride salt)
Physical state-color-odor: both the dichloride and the dimethyl sulfate salts
are white deliquescent crystalline solids; the
technical product is greater than 35% pure
Melting point: both salts - decompose about 300°C
Solubility: both salts are freely soluble in water, sparingly soluble in
lower alcohols, insoluble in hydrocarbons
Stability: both salts are stable under acid conditions but are hydrolyzed by
alkali; generally compatible with non-alkaline aqueous solutions
but may be inactivated by inert clays and anionic surfactants;
decompose in UV light; unformulated products are corrosive to
metals
Other names: Crisquat; Dextrone X; Dexuron; Esgram; Gramonol; Gramoxone;
Gramuron; Herboxone; Para-Col; Pathclear; Pillarquat;
Pillarxone; Sweep; Terraklene; TotaCol; Toxer Total; Weedol
-------
3rd Update - August 1982 Paraquat EPA-1
Reagents;
1. Paraquat (dichloride) standard of known purity
2. Phenol (internal standard) of known purity (make sure that the phenol
gives a clean chroma tog ram with no co-eluting peaks)
3. Aqueous mobile phase - (0.0025M 1-heptane sulfonic acid sodium salt and
0.04M tetramethylammonium chloride adjusted to
pH 3-0 with sulfuric acid) Filter through a 0.45
micron fiIter.
4. Organic mobile phase - (0.06M tetramethylammonium chloride in 200 ml water
adjusted to pH 3.0 with sulfuric acid plus 800 ml
acetonitrile) Filter through a 0.45 micron filter.
5. Internal standard solution - weigh 1.5 grams phenol into a 500 ml
volumetric flask, dissolve in and make to volume with aqueous mobile
phase, (cone 3 mg/ml )
Equipment;
1. High Performance Liquid Chromatograph with UV detector at 254 nm. If a
variable wavelength detector is available, other wavelengths may be used
to increase sensitivity or to eliminate interference.
2. Column: MicroPak MCH-10 (30 cm x 4 mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 80% aqueous mobile phase + 2Q% organic phase
Column temperature: ambient (for excessive tailing, increase organic mobile
phase or insulate column at 25 C)
Flow rate: 1.5 to 2.0 ml/min
Wavelength: 254 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 50 mg paraquat (dichloride) standard into a 100 ml volumetric flask,
dissolve in and make to volume with internal standard solution; mix well.
Filter a portion through a 0.45 micron filter, (cone 0.5 mg paraquat
(dichloride) and 3 mg phenol per ml)
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3rd Update - August 1982 Paraquat EPA-1
Preparation of sample;
Weigh a portion of sample equivalent to 50 mg paraquat (dichloride) into a
125 ml screw-cap flask, add 100 ml internal standard solution by pipette,
close tightly, and shake on a mechanical shaker for 30 minutes. Filter a
portion through a 0.^5 micron filter, (cone 0.5 mg paraquat (dichloride) and
3 mg phenol per ml)
HPLC Determination!
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample solutions.
Calculat ion;
Measure the peak heights or areas of the paraquat (dichloride) and the phenol
for both the standard and sample solutions and calculate the following ratios:
peak height or area paraquat (dichloride)
Ratio of standard = ——;—^-. —.——,
peak height or area phenol
_ , peak height or area paraquat (dichloride)
Ratio of sample = - - - - - -
peak height or area phenol
Average the standard and sample ratios, and calculate the percent paraquat
(dichloride) as follows:
(ratio of sample) (weight of standard) (% purity of standard)
(ratio of standard)(weight of sample)
Method submitted by EPA - NEIC, Denver, Colorado (G. Thomas Gale)
January 1980
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Pendimethalin EPA-1
Determination of Pendimethalin by Gas Chromatography (TCD-IS)
Pendimethalin is the accepted (ANSI, WSSA) common name for N-(1-
ethylpropyl)-3,^-dimethyl-2,6-dinitrobenzeamine, a registered herbicide having
the chemical structure:
NO,
-CH
CH,
Molecular formula:
Molecular weight: 281.3
Physical state-color-odor: orange-yellow crystalline solid with a faint nutty
odor
Melting point: 5^ to 58°C
Boiling point: 330°C
Solubility: less than 0.5 ppm in water at 20°C; soluble in chlorinated
hydrocarbons and aromatic solvents
Stability: stable to alkaline and acidic conditions; non-corrosive
Other names: AC 92553; Accotab; Cynoff; Go-Go-San; Herbadox; Nicocyan; Pay-
off; penoxyn; penoxalin; Prowl; N-(1-ethyIpropyl)-2,6-dinitro-
3,^-xylidine
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3rd Update - August 1982 Pendimethalin EPA-1
Reagents;
1. Pendimethalin standard of known purity
2. Pyrene (internal standard) of known purity
Note: pyrene is available from Sigma Chemical Co. and Aldrich Chemical
Co. Pyrene is a possible carcinogen and should be handled accordingly.
3. Chloroform, pesticide grade
k. Internal standard solution - weigh 1.0 gram pyrene into a 100 ml
volumetric flask, dissolve in and make to volume with chloroform; mix
wel1. (cone 10 mg/ml)
Equi pment;
1. Gas chromatograph with thermal conductivity detector (TCD)
2. Column: 4 ' x 1/V glass packed with 3-8$ SE-30 on 80 to 100 mesh
Diatoport S (or equivalent column)
3. Precision liquid syringe
k. Usual laboratory glassware
Operating conditions for TCD:
Column temperature: 205°C
Injection port temperature: 225°C
Detector temperature: 230°C
Fi lament current: 200 ma
Carrier gas: Helium - flow adjusted as necessary
Procedure;
(This method is for emulsifiable concentrates but could be adapted to other
formulations if and when they become available.)
Preparation of standard;
Weigh 140 mg pendimethalin standard into a small screw-cap flask or bottle,
add 20 ml internal standard solution by pipette and 20 ml chloroform by
graduated cylinder (or pipette); mix thoroughly, (cone 3-5 mg pendimethalin
and 5 mg pyrene per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 1^0 mg pendimethalin into a small
flask or bottle and treat exactly as under preparation of standard as above,
Shake thoroughly to insure adequate mixing of sample and solvent.
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3rd Update - August 1982 Pendimethalin EPA-1
GC Determination;
Inject 3 to 5 ul of standard and, if necessary, adjust the instrument
parameters and the volume injected to give a complete separation within a
reasonable time and to obtain peak heights of 1/3 to 2/3 full scale. Proceed
with the determination, making at least three injections each of standard and
sample solutions. The elution order is pendimethalin then pyrene.
Calculat ion;
Measure the peak heights or areas of the pendimethalin and pyrene for both the
standard and sample solutions and calculate the following ratios;
peak height or area pendimethal i n
Ratio of standard = —:—;—^
peak height or area pyrene
peak height or area pendimethalin
Ratio of sample
peak height or area pyrene
Average the standard and sample ratios, and calculate the percent
pendimethalin as follows:
(ratio of sample) (weight of standard) (% purity of standard)
(ratio of standard) (weight of sample)
Method submitted by EPA (former) Product Analysis Laboratory, Region II,
New York, NY
June 1977
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Pentachlorophenol EPA-1
Determination of Pentachlorophenol by Gas Chromatography (FID-IS)
Using on Column Derivatization with MSFTA
Pentachlorophenol (also commonly known as PCP and penta) is a registered
insecticide, fungicide, herbicide, and molluscicide. It has the chemical
structure:
Cl
Cl
Molecular formula: CgHC1,-0
Molecular weight: 266.34
Physical state-color-odor: colorless crystals with a phenolic odor (crude
products are dark grayish flakes or powder)
Melting point: 191°C (pure); 18? to 189°C (crude)
Solubility: 20 ppm in water at 30 C; soluble in most organic solvents;
limited solubility in carbon tetrachloride and petroleum oils of
low aromatic or olefin content
Stability: non-flammable; non-corrosive except in presence of moisture;
aqueous solutions have an alkaline reaction
Sodium salt: forms buff colored flakes with one mole of water of
crystallization; solubility in water is 33 grams/100 ml at 25°C;
insoluble in petroleum oils
Other names: Antimicrobial; Dowicide; Dowicide G; Dowicide EC-7; Dow
Pentachlorophenol DP-2; penchlorol; Pentacon; Penwar; Priltox;
Santobrite; Santophen; Sinituho; Weedone
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3rd Update - August 1982 Pentachlorophenol EPA-1
Reagents;
1. Pentachlorophenol standard of known purity
2. Dibutyl phthalate (internal standard) of known purity
3. Acetone, analytical grade or better
4. MFSTA [N-methyl-N-trimethyl-sllyltrifluoracetamide] derivatization reagent
5. Internal standard solution - weigh 500 mg dibutyl phthalate into a 100 ml
volumetric flask, dissolve in and make to volume with acetone, and mix
wel1. (cone 5 mg/ml)
Equipment;
1. Gas chromatograph with flame ionization detector
2. Column: 6' x 1/V glass packed with 3% OV-1 on Supelcoport 100/200 (or
equivalent column)
3. Precision liquid syringe: 10 ul
k. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions for FID;
Column temperature: 180°C
Injection port temperature: 225°C
Detector temperature: 300°C
Carrier gas: nitrogen - flow adjusted as necessary (approx. 25 ml/min)
Hydrogen flow: adjusted as necessary (approx. 30 ml/min)
Air flow: adjusted as necessary (approx. 800 ml/min)
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure;
This method is for liquid formulations but could easily be adapted for solid
formulations such as dusts, granules, powders, etc.
Preparation of standard;
Weigh 75 rng pentachlorophenol standard into a 50 ml volumetric flask, add
10 ml internal standard solution, and make to volume with acetone; mix well
(cone 1.5 mg pentachlorophenol and 1 mg dibutyl phthalate per ml)
-------
3rd Update - August 1982 Pentachlorophenol EPA-1
Preparation of sample;
For liquid samples, weigh a portion of sample equivalent to 75 mg pentachloro-
phenol into a 50 ml volumetric flask, add 10 ml internal sample, and make to
volume with acetone; mix well, (cone as above)
For granular or solid samples, weigh a portion of sample equivalent to 75 mg
pentachlorcphenol into a small (100/125 ml) screw-cap flask or bottle, add
10 ml internal standard solution and 40 ml acetone by pipette, close tightly
and shake for 30 minutes on a mechanical shaker. Allow to settle, centrifuge
or filter if necessary to obtain a clear solution, (cone as above)
GC Determination:
Using a 10 ul syringe, fill as follows: 1 ul acetone, 1 ul air, 1 ul MSTFA,
and 2 ul standard (or sample) solution. Make an injection of standard and, if
necessary, adjust the instrument parameters and the volume injected (keep the
same relative amounts as above) to give a complete separation within a
reasonable time and to obtain peak heights of 1/2 to 3/4 full scale. Proceed
with the determination, making at least three injections of sample - each
preceeded and followed by an injection of standard.
Calculat ion;
Measure the peak heights or areas of the pentachlorophenol and dibutyl
phthalate for both the standard and sample solutions and calculate the
following ratios:
peak heights or area pentachlorophenol
ratio of standard = —-—•—~ :-; ;—. . ,
peak heights or area dibutyl phthalate
peak heights or area pentachlorophenol
ratio of sample = •—-—— : —•—•—
peak heights or area dibutyl phthalate
Average the standard and sample ratios, and calculate the percent penta-
chlorophenol as follows:
_ (ratio of sample) (weight standard) (% purity of standard)
(ratio of standard)(weight sample)
Method submitted by EPA - NEIC, Denver, Colorado (Chuck Rzeszutko)
August 1979
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
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3rd Update - August 1982
Pentachlorophenol EPA-2
Determination of Pentachlorophenol by High Performance Liquid Chromatography
Pentachlorophenol (also commonly known as PCP and penta) is a registered
insecticide, fungicide, herbicide, and molluscicide. It has the chemical
structure:
Cl
Molecular formula:
Molecular weight: 266.3^
Physical state-color-odor: colorless crystals with phenolic odor (crude
products are dark grayish flakes or powder)
Melting point: 191°C (pure); 18? to 189°C (crude)
Solubility: 20 ppm in water at 30 C; soluble in most organic solvents;
limited solubility in carbon tetrachloride and petroleum oils of
low aromatic or olefin content
Stability: non-flammable; non-corrosive except in the presence of moisture;
aqueous solutions have an alkaline reaction
Sodium salt: forms buff colored flakes with one mole of water of
crystallization; solubility in water in 33 grams/100 ml at 25°C;
insoluble in oetroleum oils
Other names: Antimicrobial; Dowicide; Dowicide G; Dowicide EC-7; Dow
Pentachlorophenol DP-2; penchlorol; Pentacon; Penwar; Priltox;
Santobrite; Santophen; Sinituho; Weedone
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3rd Update - August 1982 Pentachlorophenol EPA-2
Reagents;
1. Pentachlorophenol standard of known purity
2. Benzyl benzoate (internal standard) of known purity
3. Methanol/PIC A - (1 bottle PIC A in one liter of 3$% methanol + 10% water
filtered through a 0.45 micron filter)
4. Water/PIC A - (1 bottle PIC A in one liter of water filtered through a
0.045 micron fi1ter)
5. Internal standard solution - weigh 250 mg benzyl benzoate into a 500 ml
volumetric flask, dissolve in and make to volume with methanol/PIC A
solution, (cone 0.5 mg/ml)
Equipment:
1. High Performance Liquid Chromatograph with a variable wavelength UV
detector at 218.5 nm. If a variable wavelength detector is not available,
operating parameters and concentrations may have to be changed to obtain
the necessary separation and sensitivity
2. Column: MicroPak MCH-10 (30 cm x 4 mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 70S (90? MeOH/10% water/PIC A) + 30% (water/PIC A)
Column temperature: 32°C
Flow rate: 2 ml/min
Wavelength: 218.5 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg pentachlorophenol into a screw-cap flask, add 100 ml internal
standard solution by pipette, and shake to dissolve. Dilute 5 ml to 50 ml
with internal standard solution and filter a portion through a 0.45 micron
filter, (cone 0.1 mg pentachlorophenol and 0.5 mg benzoate per ml)
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3rd Update - August 1982 Pentachlorophenol EPA-2
preparation of sample;
Weigh a portion of sample equivalent to 100 mg pentachlorophenol into a 125 ml
screw-cap flask, add 100 ml internal standard solution by pipette, close
tightly, and shake for a few minutes then place in an ultrasonic bath about 5
minutes. Shake again for a few minutes and dilute 5 ml to 50 ml with internal
standard solution. Filter a portion through a 0.^5 micron filter.
(cone as above)
HPLC Determination:
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making aleternately three injections
each of standard and sample solutions.
Calculat ion;
Measure the peak heights or areas of the pentachlorophenol and the benzyl
benzoate for both the standard and sample solutions and calculate the
following ratios:
peak height or area pentachlorophenol
Ratio of standard = —: r- r \
peak height or area benzyl benzoate
peak height or area pentachlorophenol
Ratio of sample =-
peak height or area benzyl benzoate
Average the standard and sample ratios, and calculate the percent penta-
chlorophenol as follows:
(ratio of sample) (weight standard) (% purity of standard)
(ratio of standard) (weight sample)
Method submitted by EPA - NEIC, Denver, Colorado (Phil Gee and G. Thomas Gale)
January 1980
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982 Propionic Acid EPA-1
Determination of Propionic Acid by Gas Chromatography (FID)
Propionic acid is a registered fungicide having the chemical structure:
CH3 CH2 COOH
Molecular formula: C-HgO-
Molecular weight: 74.08
Physical state-color-odor: oily liquid with a slightly pungent, disagreeable,
ranc i d odor
Melting point: -21.5°C
Boiling point: 141.1°C at ?60 mm Hg
Solubility: miscible with water; soluble in alcohol, ether, chloroform; can
be salted out of water solutions by the addition of calcium
chloride or other salts
Stability: stable - (example: propionates are used as mold inhibitors in
bread; blends of acetic acid and propionic acid are used as liquid
grain preservatives)
Other names: ChemStor; Grain Treat; propanoic acid; Propionic Acid Grain
Preserver; Sentry Grain Preserver
Reagents;
1. Propionic acid of known purity
2. Ethanol, pesticide grade
Equi pment;
1. Gas Chromatograph with flame ionization detector (FID)
2. Column: 6" x 1/V glass packed with Porapak Q
3. Precision liquid syringe: 10 ul
-------
3rd Update - August 1982 Propionic Acid EPA-1
4. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions:
Column temperature: 200°C
Injection port temperature: 250°C
Detector temperature: 250°C
Carrier gas: Helium (or nitrogen) flow - adjusted as necessary
Hydrogen flow: adjusted as necessary
Air flow: adjusted as necessary
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg propionic acid standard into a 50 ml volumetric flask, dissolve
in and make to volume with ethanol; mix well, (cone 2 mg/ml)
Preparation of sample;
Weigh a portion of sample equivalent to 100 mg propionic acid into a 125 ml
screw-cap flask, add 50 ml ethanol by pipette, close tightly and shake on a
mechanical shaker for 30 to 40 minutes. Centrifuge or filter if necessary to
obtain a clear solution, (cone 2 mg/ml)
GC Determination;
Inject 2 ul of standard and, if necessary, adjust the instrument parameters
and the volume injected to give a complete separation within a reasonable time
and to obtain peak heights of 1/2 to 3/4 full scale. Proceed with the
determination, making at least three injections each of standard and sample
solutions.
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3rd Update - August 1982 Propionic Acid EPA-1
Calculat ion;
Measure the peak height or peak area for each peak and calculate the average
for both standard and sample. Using these averages, calculate the percent
Propionic acid as follows:
(peak height or area sample)(weight standard injected)(!fc purity standard)
(peak height or area standard)(weight sampleinjected)
Method submitted by Martin J. Byrne, EPA, Region XIII, Denver, Colorado
January
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
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3rd Update - August 1982 Pyrethrins EPA-4
Determination of Pyrethrins by High Performance Liquid Chromatography
For description, structure, and technical data on pyrethrins, see
Pyrethrins EPA-1 .
Reagents;
1. Pyrethrin standard of known purity
2. Acetonitrile, HPLC grade
3. Water, HPLC grade
Equipment:
1. High Performance Liquid Chroma tograph with UV detector at 25^ nm. If a
variable wavelength detector is available, other wavelengths may be used
to increase sensitivity or to eliminate interference.
2. Column: MicroPak MCH-10 (30 cm x 4 mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.^5 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 75% acetonitrile + 25% water
Column temperature: ambient
Flow rate: 1.5 ml/min
Wavelength: 254 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg pyrethrin standard into a 125 ml screw-cap flask, add 100 ml
acetonitrile by pipette, close tightly and shake to dissolve. Filter through
a 0.^5 micron filter, (cone 1 mg/ml)
-------
3rd Update - August 1982 Pyrethrins EPA-4
Preparation of sample:
Weigh a portion of sample equivalent to 100 mg pyrethrin into a 125 ml screw-
cap flask, add 100 ml acetonitrile by pipette, close tightly, and shake for 30
minutes. (A few minutes in an ultrasonic bath may help effect solution)
Filter through a 0.45 micron filter, (cone 1 mg/ml)
HPLC Determination;
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample solutions.
Calculat ion:
Measure the peak height or peak area for each peak and calculate the average
for both standard and sample. Using these averages, calculate the percent
pyrethrins as follows:
(peak height or area sample)(weight standard injected)($ purity standard)
(peak height or area standard)(weight sample injected)
Method submitted by EPA - NEIC, Denver, Colorado (G. Thomas Gale)
January 1980
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
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3rd Update - August 1982
Rotenone EPA-2
Determination of Rotenone by High Performance Liquid Chromatography
Rotenone is the trivial names of the main insecticidal component of
certain species of "Derris" and "Lonchocarpus". It is a registered
insecticide (with some acaricidal properties) and has the chemical structures
0—CH
Molecular formula: ^23^22^6
Molecular weight: 391*.1*
Physical state-color-odor: colorless crystals; crystallizes with solvent of
crystal 1ization
Melting point: 163°C ( a dimorphoric form melts at 181°C)
Solubility: 15 ppm in water at 100°C; slightly soluble in petroleum oils and
carbon tetrachloride; soluble in polar organic solvents
Stability: readily oxidized especially in presence of light or alkali
Other names: barbasco; Chem Fish; cube'; derris; haiari; neko; nicouline;
Prentox; tubatoxin
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3rd Update - August 1982 Rotenone EPA-2
Reagents;
1. Rotenone standard of known purity
2. Acetonitrile, HPLC or pesticide grade
3. Methanol, HPLC or pesticide grade
Equipment;
1. High Performance Liquid Chromatograph with a variable wavelength UV
detector at 295 nm. If a variable wavelength detector is not available,
operating parameters and concentrations may have to be changed to obtain
the necessary separation and sensitivity.
2. Column: uBondapak C18 (30 cm x 3-9 mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 45% acetonitrile + 55$ water
Column temperature: 33°C
Flow rate: 3 ml/min
Wavelength: 295 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg rotenone standard into a 125 ml screw-cap flask, add 100 ml
methanol by pipette, close tightly, and shake to dissolve. Filter a portion
through a 0.45 micron filter, (cone 1 mg/ml)
Preparation of sample;
Weigh a portion of sample equivalent to 100 mg rotenone into a 125 ml screw-
cap flask, add 100 ml methanol by pipette, close tightly, and shake several
minutes. Place in an ultrasonic bath for 2 or 3 minutes and then shake on a
mechanical shaker for one hour. Filter a portion through a 0.45 micron
filter, (cone 1 mg/ml)
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3rd Update - August 1982 Rotenone EPA-2
HPLC Determination!
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
tims. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample solutions.
Calculation;
Measure the peak height or area for each peak and calculate the average for
both standard and sample. Using these averages, calculate the percent
rotenone as follows:
(peak height or area sample)(weight standard injected)(% purity standard)
(peak height or area standard) (weight sample injected)
Method submitted by EPA - NEIC, Denver, Colorado (Chuck Rzeszutko)
November 1979
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Strychnine EPA-4
Determination of Strychnine by High Performance Liquid Chromatography
Strychnine is an alkaloid extracted from the seeds of "Strychnos nux-
vomica". it is a registered rodentrcide and has the chemical structure:
CH0 0 — CH,
CH
CH
0
H N /CH C
^C CH ^CH
CH
.
CH
CH,
CH,
CH
CH
•N
CH,
Molecular formula: ^21^22^2^2
Molecular weight: 33^.A
Physical state-color-odor: hard white crystals or powder; very bitter taste;
very poi sonous!
Melting point: 270 to 280°C with decomposition
Solubility: Practically insoluble in water, cold alcohol and cold ether;
slightly soluble in benzene and chloroform
Stability: forms salts with acids; precipitated by alkaloid precipitants
Other names: Kwik-kil; Mouse-tox; Ro-Dec
Strychnine hydrochloride: colorless prisms containing water of
crysti11ization (1 1/2 - 2 mol), lost at 100°C;
soluble in water and alcohol; insoluble in ether
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3rd Update - August 1982 Strychnine EPA-4
Strychnine sulfate: white crystallization powder containing water of
crystallization lost at 110°C, mp above 199°C; moderately
soluble in water and alcohol; insoluble in ether
Reagents;
1. Strychnine standard of known purity
2. Phenol (internal standard) of known purity (make sure that the phenol
gives a clean chromatogram with no co-eluting peaks)
3. Aqueous mobile phase - (0.0025M 1-heptane sulfuric acid sodium salt and
0.04M tetramethylammonium chloride adjusted to
pH 3.0 with sulfuric acid) Filter through a 0.45
micron fiIter .
4. Organic mobile phase - (0.06M tetramethylammonium chloride in 200 ml water
adjusted to pH 3-0 with sulfuric acid plus 800 ml
acetonitrile) Filter through a 0.45 micron filter.
5. Internal standard solution - weigh 1.125 grams phenol into a 500 ml
volumetric flask, dissolve in and make to volume with aqueous mobile
phase, (cone 2.25 mg/ml)
Equipment;
1. High Performance Liquid Chroma tograph with UV detector at 254 nm. If a
variable wavelength detector is available, other wavelengths may be used
to increase sensitivity or to eliminate interference.
2. Column: MicroPak MCH-10 (30 cm x 4 mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 30% aqueous mobile phase + 70% organic mobile phase
Column temper3ture: ambient (may be somewhat sensitive to temperature
changes)
Flow rate: 1.5 ml/min
Wavelength: 254 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure;
Preparation of standard;
Weigh 87.5 mg strychnine standard into a 50 ml volumetric flask, dissolve in
and make to volume wi th interna 1 standard solut ion; mix wel1. Dilute 10 ml to
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3rd Update - August 1982 Strychnine EPA-4
50 ml with internal standard solution and mix well. Filter a portion through
a 0.45 micron filter, (cone 0.35 mg strychnine and 2.25 mg phenol per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 35 mg strychnine into a 125 ml screw-
cap flask, add 100 ml internal standard solution by pipette, close tightly,
place in an ultrasonic bath for several minutes, then shake on a mechanical
shaker for one hour. Filter a portion through a 0.45 micron filter, (cone
0.35 mg strychnine and 2.25 mg phenol per ml)
HPLC Determination;
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample solutions. Elution order is strychnine then
phenol .
Calculation;
Measure the peak heights or areas of the strychnine and the phenol for both
standard and sample solutions and calculate the following ratios:
peak height or area strychnine
Ratio of standard = —;—r—, ;
peak heignt or area phenol
peak height or area strychnine
Ratio of sample =
peak height or area phenol
Average the standard and sample ratios, and calculate the percent strychnine
as follows:
(ratio of sample) (weight standard) (% purity of standard)
(ratio of standard)(weight sample)
Method submitted by EPA - NEIC, Denver, Colorado (G. Thomas Gale)
March 1980
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Tebuthiuron EPA-1
Determination of Tebuthiuron by Ultraviolet Spectroscopy
Tebuthiuron is the accepted (ANSI, BSI, WSSA) common name for
N-[5-(1,1-dimethylethyl)-1,3,4-thiadiazol-2-yl]N,N-dimethylurea, a registered
herbicide having the chemical structure:
N-
CH,
CH,. C-
CH,
-N
CH,
-N—C—N—CH,
II I
0 H
Molecular formula:
Molecular weight: 228.31
Physical state-color-odor: colorless, odorless solid
Melting point: 161.5 to 164 C
Solubility: grams per 100 ml solvent at 25°C: chloroform - 25;
methanol - 17; acetone - 7; acetonitrile - 6; methyl
cellosolve - 6; hexane - 0.6; benzene - 0.37; water - 0.23
Stability: stable under usual storage conditions; non-corrosive; non-
flammable; generally compatible with most other herbicides
Other names: EL-103; Graslan; Spike; 1-(5-tert-butyl-1,3,^-thiadiazol-2-yl)
1,3-dimethylurea
Reagents;
1. Tebuthiuron standard of known purity
2. Methanol, pesticide or spectro grade
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3rd Update - August 1982 Tebuthiuron EPA-1
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording with matched
1 cm eel Is .
2. Mechanical shaker
3. Centrifuge or filtration apparatus
k. Usual laboratory glassware
Procedure:
Preparation of standard;
Weigh 80 mg tebuthiuron standard into a 100 ml volumetric flask, dissolve in
and make to volume with methanol . Mix thoroughly and pipette a 10 ml aliquot
into a second 100 ml volumetric flask, make to volume with methanol and mix
thoroughly. Pipette 10 ml into a third 100 ml volumetric flask, make to
volume with methanol and mix thoroughly, (final cone 8 ug tebuthiuron/ml).
Preparation of sample;
Weigh a portion of sample equivalent to 80 mg tebuthiuron into a 250 ml glass-
stoppered flask or screw-cap bottle, add 100 ml methanol by pipette, close
tightly and shake on a mechanical shaker for 30 minutes. Allow to settle,
centrifuge or filter if necessary, taking precautions to avoid evaporation of
solvent. Dilute 10 ml to 100 ml and then 10 ml to 100 ml as under standard
preparation, (final cone 8 ug tebuthiuron/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative settings for the
particular instrument being used, balance the pen at 0 and 100% transmission
at 253 nm with methanol in each cell. Scan both standard and sample solutions
from 320 to 220 nm with methanol in the reference cell. Measure the
absorbance of standard and sample solutions at 253 nm.
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3rd Update - August 1982 Tebuthiuron EPA-1
Calculations;
From the above absorbances and using the standard and sample concentrations,
calculate the percent tebuthiuron as follows:
(abs. sample) (cone. std. in ug/ml) (% purity)
(abs. std.) (cone, sample in ug/ml)
There is a straight line relationship between absorbance and concentration for
up to 10 ug/ml.
Method submitted by EPA (former) Product Analysis Laboratory, Region II.
New York, NY
November 1976
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Tecnazene EPA-1
Determination of Tecnazene by Gas Chromatography (FID-IS)
Tecnazene is the accepted (BSI, ISO) common name for 1,2,4,5-tetrachloro-
3-nitrobenzene, a registered fungicide and plant growth regulator having the
chemical structure:
Cl
Molecular formula:
Molecular weight: 260.9
Physical state-color-odor: colorless, odorless crystalline solid
Melt ing point: 99 C
Solubility: practically insoluble in water; about k% in ethanol at 25°C;
readily soluble in benzene, carbon disulfide, chloroform
Stability: appreciably volatile at room temperature
Other names: Folosan; Fusarex; TCNB; 2,3,5,6-tetrachloronitrobenzene
Reagents;
1. Tecnazene standard of known purity
2. Orthophenylphenol (internal standard), analytical grade
3. Acetone, pesticide grade
A. Internal standard solution - weigh 250 mg o-phenylphenol into a 500 ml
volumetric flask, dissolve in and make to volume with acetone; mix well
(cone 0.5 mg/ml)
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3rd Update - August 1982 Tecnazene EPA-1
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: V x 1/V glass packed with 3.8% UC-V98 on 80/100 mesh Diatoport
S (or equivalent column such as SP-2100 on Chromosorb W HP)
3. Precision liquid syringe: 10 ul
A. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Us.ual laboratory glassware
Operating conditions for FID;
Column temperature: 150 to 170°C adjusted for best time and separation
Injection port temperature: 200°C
Detector temperature: 230°C
Carrier gas: helium - flow adjusted as necessary
Hydrogen flow; 30 ml/min - adjusted as necessary
Air flow: 55 ml/min - adjusted as necessary
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 90 mg tecnazene standard into a small (100/125 ml) screw-cap flask or
bottle, add 50 ml internal standard solution by pipette, and shake to
dissolve, (cone 1.8 mg tecnazene and 0.5 mg o-phenylphenol per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 90 mg tecnazene into a small flask or
bottle, add 50 ml internal standard solution, close tightly and shake for 30
minutes on a mechanical shaker. Allow to settle, centrifuge or filter if
necessary to obtain a clear solution, (cone 1.8 mg tecnazene and 0.5 mg
o-phenylphenol per ml)
GC Determination;
Inject 3 ul of standard and, if necessary, adjust the instrument parameters
and the volume injected to give a complete separation within a reasonable time
and to obtain peak heights of 1/2 to 3A full scale. Proceed with the
determination, making at least three injections each of standard and sample
solutions. The elution order is o-phenylphenol then tecnazene.
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3rd Update - August 1982 Tecnazene EPA-1
Calculat ion;
Measure the peak heights or areas of the tecnazene and o-phenylphenol for both
the standard and sample solutions and calculate the following ratios:
peak height or area tecnazene
Ratio of standard = ,—. , ,—, r
peak height or area o-phenylphenol
peak height or area tecnazene
Ratio of sample
peak height or area o-phenylphenol
Average the standard and sample ratios, and calculate the percent tecnazene as
follows:
(ratio of sample) (weight of standard) (% purity of standard)
(ratio of standard) (weight sample)
Method submitted by EPA (former) Product Analysis Laboratory, Region II,
New York, NY
March 1976
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
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3rd Update - August 1982
Terbaci1 EPA-1
Determination of Terbaci1 by Ultraviolet Spectroscopy
T'erbacil is the accepted (ANSI , BSI , ISO, WSSA) common name for
3-tert-butyl-5-chloro-6-methyl uraci 1 , a registered herbicide having the
chemical structure:
H
Cl
! Q
N.
\_/
II
0
-C—CH,
CH.
Molecular formula:
Molecular weight: 216.5
Physical state-color-odor: odorless, white crystalline solid
Melting point: 175 to 177°C
Solubility: 710 ppm in water at 25 C; moderately soluble in methyl isobutyl
ketone, butyl acetate, xylene; highly soluble in cyclohexane,
dimethylformami de
Stability: stable to heat up to mp (below which it sublimes); non-corrosive;
non-flammable; stable in water, aqueous bases, and common organic
solvents at room temperature
Other names: DuPont Herbicide 732; Sinbar
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3rd Update - August 1982 Terbacil EPA-1
Reagents:
1. Terbacil standard of known purity
2. Chloroform, pesticide or spectro grade
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording with matched
1 cm eel Is.
2. Mechanical shaker
3. Centrifuge or filtration apparatus
4. Usual laboratory glassware
Procedure:
Preparation of standard;
Weigh 150 tng terbacil standard into a 100 ml volumetric flask, make to volume
with chloroform and mix thoroughly. Pipette a 10 ml aliquot into a second
100 ml volumetric flask, make to volume with chloroform and mix thoroughly.
Pipette 10 ml into a third 100 ml volumetric flask and make to volume with
chloroform; mix thoroughly, (final cone 15 ug terbacil/ml)
Preparation of sample;
Weigh a portion of sample equivalent to 150 mg terbacil into a 250 ml glass-
stoppered flask or screw-cap bottle, add 100 ml chloroform by pipette, stopper
tightly, and shake on a mechanical shaker for 30 minutes. Allow to settle,
centrifuge or filter if necessary, taking precaution to avoid evaporation of
solvent. Dilute 10 ml to 100 ml and then 10 to 100 ml as under sample
preparation, (final cone 15 ug terbacil/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative settings for the
particular instrument being used, balance the pen at 0 and 100? transmission
at 275 nm with chloroform in each cell. Scan both standard and sample
solutions from 320 to 230 nm with chloroform in the reference cell. Measure
the absorbance of standard and sample solutions at 275 nm.
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3rd Update - August 1982 Terbaci1 EPA-1
Calculat ions;
From the above absorbances and using the standard and sample concentrations,
calculate the percent terbaci1 as follows:
(abs. sample) (cone. std. in ug/ml) (% purity)
(abs. std.)(cone, sample in ug/ml)
There is a straight line relationship between absorbance and concentration for
up to 2.') ug/ml .
Method submitted by EPA (former) Product Analysis Laboratory, Region II,
New York, NY
November 1976
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Tetrachlorvinphos EPA-1
Determination of Tetrachlorvinphos by Gas Chromatography (FID-IS)
Tetrachlorvinphos is the accepted (BSI,ISO) common name for (cis or Z
isomerof) 2-chloro-1-(2,4 ,5-trichlorophenyl) vinyl dimethyl phosphate, a
registered insecticide having the chemical structure:
Molecular formula: C.-hLCUO.P
Molecular weight: 365.96
Physical state-color-odor: off-white crystalline solid
Melting point: technical Gardona (98% minimum cis isomer) 97 to 98°C
technical Rabon (94$ by weight active ingredient) 93 to 98°C
Solubility: at 20°C: 11 ppm in water, less than 20% w/w in acetone, 40$ w/w
in chloroform and methylene chloride, less than 15$ w/w
in xyl ene
Stability: stable to 100°C; slowly hydrolyzed by neutral or acid media, more
rapidly hydrolyzed in alkaline media
Other names: Appex; CVMP; Dust M; Gardcide; Gardona; Rabon; Rabone; ROL;
SD 8447; Stirofos
Reagents;
1. Tetrachlorvinphos standard of known purity
2. n-Docosane (internal standard), practically grade (or better)
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3rd Update - August 1982 Tetrachlorvinphos EPA-1
3. Methylene chloride, pesticide grade
k. Internal standard solution - weigh 675 mg n-docosane into a 250 ml
volumetric flask, dissolve in and make to volume with methylene chloride;
mix well, (cone 2.7 mg/ml)
Equipment;
1. Gas chromatograph with a flame ionization detector (FID)
2. Column: 6' x 1/V glass packed with 10% OV-1 on 80/100 Chromosorb W
3. Precision liquid syringe: 10 ul
k. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions for F[D;
Column temperature: 225°C
Injection port temperature: 235°C
Detector temperature: 2^0°
Carrier gas: nitrogen - flow adjusted as necessary
Hydrogen flow: adjusted as necessary
Air flow: adjusted as necessary
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure;
Preparation of standard;
Weigh 200 mg tetrachlorvinphos standard into a 125 ml screw-cap flask, add
50 ml internal standard solution by pipette, and shake to dissolve; mix
thoroughly, (cone A mg tetrachlorvinphos and 2.7 nig n-docosane per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 200 mg tetrachlorvinphos into a 125 ml
screw-cap flask, add 50 ml internal standard solution by pipette, close
tightly, and shake on a mechanical shaker for one hour. Allow to settle,
centrifuge or filter if necessary to obtain a clear solution.
(cone 4 mg tetrachlorvinphos and 2.7 mg n-docosane per ml)
GC Determinat ion;
Inject 3 ul of standard and, if necessary, adjust the instrument parameters
and the volume injected to give a complete separation within a reasonable time
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3rd Update - August 1982 Tetrachlorvinphos EPA-1
and to obtain peak heights of 1/2 to 3A full scale. Proceed with the
determination, making at least three injections each of standard and sample
solutions. The elution order is tetrachlorvinphos then n-docosane.
Calculat ion;
Measure the peak heights or areas of the tetrachlorvinphos and the n-docosane
for both the standard and sample solutions and calculate the following ratios:
peak height or area tetrachlorvinphos
Ratio of standard = L . L -
peak height or area n-docosane
peak height or area tetrachlorvinphos
Ratio of sample
peak height or area n-docosane
Average the standard and sample ratios, and calculate the percent tetra-
chlorvinphos as follows:
(ratio of sample) (weight of standard) (% purity of standard)
(ratio of standard) (weight of sample)
Method submitted by E. S. Greer, EPA (formerly) Product Analysis Laboratory,
Region IX, San Francisco, CA (Mr. Greer is now at Beltsville, MD)
August 1977
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Thiophanate EPA-1
Determination of Thiophanate by Ultraviolet Spectroscopy
Thiophanate is the accepted (BSI, ISO) common name for diethyl [ 1,2-
phenylene bis (iminocarbonothioyl )] bi s [carbamate ], a registered fungicide
having the chemical structure:
NH C NH C 0 CH0 CH
NH C NH C 0 CH0 CH
Molecular formula:
Molecular weight: 370.4
Physical state-color-odor: colorless, crystalline solid
Melting point: 190 C with decomposition
Solubility: very slightly soluble in water; soluble in acetone, methanol,
chloroform, acetonitrile, cyclohexane, ethyl acetate
Stability: stable in neutral or acidic aqueous solutions, but unstable in
alkaline aqueous solutions; forms chelates with calcium, copper,
and sodium ions
Other names: 3336; Cercobin; NF-35; Topsin; Topsin E; 1,2-bis
(3-ethoxycarbonyl-2-thioureido) benzene; diethyl 4,4- o-
phenylenebis[3-thioallophanate]; thiophanate-ethyl
Analytical method: see thiophanate-methyl and use the same method
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
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3rd Update - August 1982
Thiophanate-methyl EPA-1
Determination of Thiophanate-methyl by Ultraviolet Spectroscopy
Thiophanate-methyl Is the accepted (ANSI, BSI, ISO) common name for
dimethyl [(1,2-phenylene) bis-(iminocarbonothioyl)] bis [carbamate], a
registered fungicide having the chemical structure:
-0 CH,
-0 CH,
s
/^\ »
r l
.\
, NH- — C
_ NH , r
0
II
NH C
MU r-
0
Molecular formula: ^i?^]k®k^^2
Molecular weight: 3^*2. k
Physical state-color-odor: colorless, crystalline solid; odorless to slightly
sulf urous
Melting point: 178°C (decomposes)
Solubility: practically insoluble in water; slightly soluble in common
organic solvents
Stability: stable in solid state when kept below 160°C; stable in acid
aqueous solutions, slowly decomposes in neutral aqueous solutions,
rapidly decomposes in alkaline aqueous solutions
Other names: Cercobin-M; Fungitox; Labi lite; Mil do thane; Sigma; Topsin-M
Reagents;
1. Thiophanate-methyl standard of known purity
2. Chloroform, pesticide or spectro grade
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3rd Update - August 1982 Thiophanate-methyl EPA-1
Equipment;
1. Ultraviolet spectrophotometer, double beam ratio recording with matched
1 cm eel Is
2. Mechanical shaker
3. Centrifuge or filtration apparatus
k. Usual laboratory glassware
Procedure:
Preparation of standard;
Weigh 80 mg thiophanate-methyl standard into a 100 ml volumetric flask,
dissolve in and make to volume with chloroform. Mix thoroughly, and pipette
10 ml into a second 100 ml volumetric flask. Make to volume with chloroform
and mix thoroughly. Pipette 10 ml into a third 100 ml volumetric flask, make
to volume with chloroform, and mix thoroughly.
(final cone 8 ug/ml).
Preparation of sample;
Weigh a portion of sample equivalent to 80 mg thiophanate-methyl into a 250 ml
glass-stoppered flask or screw-cap bottle, add 100 ml chloroform by pipette,
and shake on a mechanical shaker for one hour. Allow to settle, centrifuge or
filter if necessary, taking precautions to avoid evaporation of solvent.
Dilute 10 ml to 100 ml and then 10 ml to 100 ml as under sample preparation.
(final cone 8 ug thiophanate-methyl/ml)
UV Determination;
With the UV spectrophotometer at the optimum quantitative settings for the
particular instrument being used, balance the pen st 0 and 100? transmission
at 269 nm with chloroform in each cell. Scan both standard and sample
solutions from 360 to 230 nm with chloroform in the reference cell. Measure
the absorbance of standard and sample solutions at 269 nm.
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3rd Update - August 1982 Thiophanate-methyl EPA-1
Calculat ions;
From the above absorbances and using the standard and sample concentrations,
calculate the percent thiophanate-methyl as follows:
a _ (abs. sample) (cone, std. in ug/ml) (% purity)
(abs. std.) (cone, sample in ug/ml)
Method submitted by EPA (former) Product Analysis Laboratory, Region II, New
York, NY
July 1975
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982 Trichlorfon EPA-3
Determination of Trichlorfon by Gas Chromatography (FID-IS)
Using on Column Derivization with BSFTA
Trichlorfon is the accepted (ISO) common name for dimethyl (2,2,2-trich-
loro-1-hydroxyethyl) phosphonate, a registered insecticide having the chemcial
structure:
CH3—0, 0 OH Cl
^^ CH C Cl
CH3—cr ci
Molecular formula: C^HgCt^P
Molecular weight: 257.^
Physical state-color-odor: white crystalline solid
Melting point: 83 to 8^°C
Solubility: 15-4 grams in 100 ml water at 25°C; soluble in benzene, ether,
ethanol, and most chlorinated solvents; slightly soluble in
petroleum oils, and in carbon tetrachloride
Stability: stable at room temperature, but is decomposed by water at higher
temperatures and at pH 5.5 to form dichlorvos
Other names: Bayer 15922; Bovinox; Briten; Cekufon; chlorofos; Ciclosom;
Crinex; Danex; Dipterex; Dylox; Equino-Aid; Leivasom;
metrifonate; Neguvon; Proxol; trichlophon; Trinex; Tugon
Reagents;
1. Trichlorfon standard of known purity
2. Benzyl benzoate (internal standard) of known purity
3. Acetone, analytical, dry
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3rd Update - August 1982 Trichlorfon EPA-3
i». BSTFA [N,0-bis(trimethylsilyl)-trifluoroacetamide] silylation reagent
for derivization
5. Internal standard solution - weigh 850 mg benzyl benzoate into a 100 ml
volumetric flask, dissolve and make to volume with acetone, and mix well.
(cone 8.5 mg/ml)
Equipment;
1. Gas chromatograph with flame ionization detector (FID)
2. Column: 6' x 2 mm ID glass packed with 3% OV-17 on Chromosorb W HP
(or other suitable column)
3. Precision liquid syringe: 10 01
4. Mechanical shaker
5. Centrifuge or filtration apparatus
6. Usual laboratory glassware
Operating conditions;
Column temperature: 150°C
Injection port temperature: 225°C
Detector temperature: 260°C
Carrier gas: nitrogen - flow adjusted as necessary
Hydrogen flow: adjusted as necessary
Air flow: adjusted as necessary
Operating parameters (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibiIity.
Procedure:
Preparation of standard;
Weigh 110 mg trichlorfon standard into a 125 ml screw-cap flask, add 10 ml
internal standard solution by pipette and *fO ml acetone by pipette, close
tightly and mix well, (cone 2.2 mg trichlorfon and 1.7 mg benzyl benzoate
per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 110 mg trichlorfon into a 125 ml
screw-cap flask, add 10 ml internal standard solution by pipette and ^0 ml
acetone by pipette, close tightly and shake for 30 minutes on a mechanical
shaker. Allow to settle, and if necessary centrifuge or filter to clarify.
(cone 2.2 mg trichlorfon and 1.7 mg benzyl benzoate per ml)
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3rd Update - August 1982 Trichlorfon EPA-3
GC Determination;
Using a 10 ul syringe, fill as follows: 1 ul acetone, 1 ul air, 1 ul BSTFA,
and 2 ul standard (or sample) solution. Make an injection of standard and, if
necessary, adjust the instrument parameters and the volume injected (keep the
same relative amounts as above) to give a complete separation within a
reasonable time and to obtain peak heights of 1/2 to 3A full scale. Proceed
with the determination, making at least three injections of sample - each
preceeded and followed by an injection of standard. Elution order is
trichlorfon then benzyl benzoate.
Calculation^
Measure the peak heights or areas of the trichlorfon and benzyl benzoate for
both the standard and sample solutions and calculate the following ratios:
peak height or area trichlorfon
ratio of standard ;—j—7—, j r—;
peak height or area benzyl benzoate
peak height or area trichlorfon
ratio of sample
peak height or area benzyl benzoate
Average the standard and sample ratios, and calculate the percent trichlorfon
as follows:
(ratio of sample) (weight standard) (% purity of standard)
(ratio of standard)(weight sample)
Method submitted by EPA - NEIC, Denver, Colorado (G. Thomas Gale)
September 1980
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982 Trichlorfon EPA-4
Determination of Trichlorfon by High Performance Liquid Chromatography
Trichlorfon is the accepted (ISO) common name for dimethyl (2,2,2-
trichloro-1-hydroxyethyl) phosphonate, a registered insecticide having the
chemical structure:
CH~ Ov 0 OH Cl
3 \ll I I
P CH C Cl
CH3 0' ci
Molecular formula: CrHnCl,0.p
Molecular weight: 257.^
Physical state-color-odor: white crystalline solid
Melting point: 83 to 8*t°C
Solubility: 15.k grams in 100 ml water at 25°C; soluble in benzene, ether,
ethanol, and most chlorinated solvents; slightly soluble in
petroleum oils, and in carbon tetrachloride
Stability: stable at room temperature, but is decomposed by water at higher
temperatures and at pH 5-5 to form dichlorvos
Other names: Bayer 15922; Bovinox; Briten; Cekufon; chlorofos; Ciclosom;
Crinex; Danex; Dipterex; Dylox; Equino-Aid; Leivasom;
metrifonate; Neguvon; Proxol; trichlorphon; Trinex; Tugon
Reagents:
1. Trichlorfon standard of known purity
2. Phenol (internal standard) of known purity (make sure that the phenol
gives a clean chromatogram with no co-eluting peaks)
3. Acetonitrile, HPLC grade
k. Water, HPLC grade
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3rd Update - August 1982 Trichlorfon EPA-4
5. Internal standard solution - weigh 125 mg phenol into a 100 ml volumetric
flask, dissolve in and make to volume with acetonitr5le; mix well. Dilute
10ml to 100 ml . (cone 0.125 rig/ml)
Equi pment;
1. High Performance Liquid Chromatograph with a variable wavelength UV
detector at 224 nm. If a variable wavelength detector is not available,
operating parameters and concentrations may have to be changed to obtain
the necessary separation and sensitivity.
2. Column: uBondapak C18 (30 cm x 3-9 mm 10) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.45 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 30fc acetonitrile + 70% water
Column temperature: 33°C
Flow rate: 2 ml/min
Wavelength: 224 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedure:
Preparation of standard;
Weigh 100 mg trichlorfon standard into a 10 ml volumetric flask, add 2 ml
internal standard solution by pipette, and make to volume with acetonitrile.
Mix thoroughly and filter a portion through a 0.45 micron filter, (cone 10 mg
trichlorfon and 0.025 mg phenol per ml)
Preparation of sample;
Weigh a portion of sample equivalent to 100 mg trichlorfon into a 10 ml
volumetric flask, add 2 ml internal standard solution by pipette, and make to
volume with acetonitrile. Stopper tightly and place in an ultrasonic bath for
10 minutes. Filter a portion through a 0.45 micron filter, (cone as above)
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3rd Update - August 1982 Trichlorfon EPA-**
HPLC Determination:
Inject 10 ul of standard solution and, if necessary, adjust the flow rate
and/or mobile phase composition to give good separation in a reasonable
time. Adjust the attenuation or the amount injected to give convenient size
peaks. Proceed with the determination making alternately three injections
each of standard and sample solutions.
Calculat ion;
Measure the peak heights or areas of the trichlorfon and the phenol for both
the standard and sample solutions and calculate the following ratios:
peak height or area trichlorfon
Ratio of standard =
peak height or area phenol
peak height or area trichlorfon
Ratio of sample =
peak height or area phenol
Average the standard and sample ratios, and calculate the percent trichlorfon
as follows:
(ratio of sample) (weight standard) (% purity of standard)
(ratio of standard)(weight sample)
Method submitted by EPA - NEIC, Denver, Colorado (G. Thomas Gale)
September 1980
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
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3rd Update - August 1982
Warfarin EPA-4
Determination of Warfarin by High Performance Liquid Chromatography
Warfarin is the accepted (BSI,ISO) common name for 3-(alpha-
acetonylbenzyl)-^-hydroxycoumarin, a registered rodenticide having the
chemical structure:
Molecular formula:
Molecular weight: 308.3
Physical state-color-odor: colorless, odorless, tasteless crystals (dl form)
Melting point: 159 to 161°C (d1 form)
Solubility: practically insoluble in water and benzene; moderately soluble in
alcohols; readily soluble in acetone and dioxane; forms water
soluble salts with sodium
Stability: stable under normal conditions
Other names: coumafene (France); zoocoumarin (Netherlands and USSR); Co-Rax;
Cov-R-Tox; Kypfarin; Ratox; RAX; Rodex; Rodex Blox; Tox-Hid;
Warfarin Plus; Warfarin Q
Reagents;
1. Warfarin standard of known purity
2. Methanol/PIC A - (1 bottle PIC A in one liter of 90£ methanol + 10% water
filtered through a 0.45 micron filter)
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3rd Update - August 1982 Warfarin EPA-**
3. Water/PIC A - (1 bottle PIC A in one liter water filtered through a 0.45
micron fiIter)
Equipment;
1. High Performance Liquid Chromatograph with a variable wavelength UV
detector at 312 nm. If a variable wavelength detector is not available,
operating parameters and concentrations may have to be changed to obtain
the necessary separation and sensitivity.
2. Column: MicroPak MCH-10 (30 cm x A mm ID) or equivalent column
3. High pressure liquid syringe or sample injection loop: 10 ul
4. Mechanical shaker and/or ultrasonic bath
5. 0.^5 micron filtering apparatus
6. Usual laboratory glassware
Operating conditions;
Mobile phase: 50$ (90%/10% water/PIC A) + 50? (water/PIC A)
Column temperature: 32°C
Flow rate: 2 ml/min
Wavelength: 312 nm
Operating conditions (above) as well as attenuation and chart speed should be
adjusted by the analyst to obtain optimum response and reproducibi1ity.
Procedu re:
Preparation of standard;
Weigh 110 mg warfarin standard into a 125 nil screw-cap flask, add 100 ml
methanol/PIC A solution by pipette, close tightly, and shake to dissolve.
Dilute 5 ml to 50 ml with methanol/PIC A solution and filter through a 0.^5
micron filter, (cone 0.11 mg/ml)
Preparation of sample;
Weigh a portion of sample equivalent to 110 mg warfarin into a 125 ml screw-
cap flask, add 100 ml methanol/PIC A solution, close tightly, and shake for
several minutes. Place in an ultrasonic bath 2 or 3 minutes, then shake again
for several minutes. Dilute 5 ml to 50 ml with methanol/PIC A solution and
filter through a 0.^5 micron filter, (cone 0.11 mg/ml)
-------
3rd Update - August 1982 Warfarin EPA-A
HPLC Determination;
Inject 10 ul of standard and, if necessary, adjust the flow rate and/or mobile
phase composition to give good separation in a reasonable time. Adjust the
attenuation or the amount injected to give convenient size peaks. Proceed
with the determination making alternately three injections each of standard
and sample solution.
Calculation;
Measure the peak height or area for each peak and calculate the average for
both standard and sample. Using these averages, calculate the percent
warfarin as follows:
(peak height or area sample)(weight standard injected)(fc purity standard)
(peak height or area standard) (weight sample injected)
Method submitted by EPA - NEIC, Denver Colorado (Phil Gee & G. Thomas Gale)
Any criticisms, suggestions, or data concerning the use of this method will be
appreciated.
-------
3rd Update - August 1982 Ziram EPA-1
Determination of Ziram by Ultraviolet Spectroscopy
Ziram is the accepted (BSI, ISO) name for zinc dimethyldithiocarbamate,
a registered fungicide having the chemical structure:
CH3, I I /CH3
>N—C —S—Zn—S—C —N'
Molecular formula: C^H^^S^Zn
Molecular weight: 305.79
Physical state-color-odor: odorless, white powder
Melting point: 2kO°C (pure), 2kO to 2M°C (technical)
Solubility: 65 ppm in water at 25°C; slightly soluble in ethanol, ether;
moderately soluble in acetone; soluble in dilute alkali,
chloroform, carbon disulfide
Stability: stable under normal conditions, but is decomposed by acids;
compatible with other pesticides except copper and mercury
compounds
Other names: Antene; Carbazinc; Corozate; Cuman; Drupina 90; Fuclasin Ultra;
Fuklasin; Fungostop; Hexazir; Mezene; Pomarsol Z Forte;
Prodaram; Tricarbamix Z; Triscabol; Vancide MZ-96; Z-C Spray;
Zerlate; Zincmate; Ziramvis; Zirasan 90; Zirberk; Zirex 90;
Ziride; Zitox
Reagents;
1. Ziram standard of known purity
2. Chloroform, pesticide or spectro grade
-------
3rd Update - August 1982 Ziram EPA-1
Equipment;
1. Ultraviolet Spectrophotometer, double beam ratio recording with matched
1 cm eel Is
2. Mechanical shaker
3. Water bath
4. Filtration apparatus with anhydrous sodium sulfate
5. Usual laboratory glassware
Procedure:
Preparation of standard;
Weigh 100 mg ziram standard into a 100 ml volumetric flask, dissolve in and
make to volume with chloroform. Mix thoroughly and pipette 10 ml into a
second 100 ml volumetric flask. Make to volume with chloroform, mix
thoroughly, and pipette 5 nil into a third 100 ml volumetric flask. Make to
volume with chloroform and mix thoroughly, (final cone 5 ug/ml).
preparation of sample (liquid-viscous formulations);
Weigh a portion of sample equivalent to 100 mg ziram into a 100 ml round-
bottom flask, add 40 ml chloroform, and reflux for 30 minutes on a boiling
water bath. Filter through anhydrous sodium sulfate into a 100 ml volumetric
flask and make to volume with chloroform. Mix thoroughly and pipette 10 ml
into a second 100 ml volumetric flask. Make to volume with chloroform, mix
thoroughly and pipette 5 ml into a third 100 ml volumetric flask; make to
volume with chloroform and mix thoroughly, (final cone 5 ug ziram/ml).
Preparation of sample (powder formulations):
Weigh a portion of sample equivalent to 100 mg ziram into a 250 ml glass-
stoppered or screw-cap flask, add 100 ml chloroform by pipette, and shake on <
mechanical shaker for 15 minutes. Allow to settle, filter and pipette 10 ml
into a 100 ml volumetric flask. Make to volume with chloroform, mix
thoroughly, and pipette 5 ml into another 100 ml volumetric flask; make to
volume with chloroform and mix thoroughly, (final cone 5 ug ziram/ml).
UV Determination;
With the UV spectrophotometer at the optimum quantitative settings for the
particular instrument being used, balance the pen at 0 and 100£ transmission
at 261 nm with chloroform in each cell. Scan both the standard and sample
solutions from 300 to 200 nm with chloroform in the reference cell. Measure
the absorbance of standard and sample at 261 nm.
-------
3rd Update - August 1982 Ziram EPA-1
Calculat ions;
From the above absorbances and using the standard and sample concentrations,
calculate the percent ziram as follows:
g _ (abs. sample) (cone, std. in ug/ml) (% purity)
(abs. std.)(cone, sample in ug/ml)
Method submitted (summer - ?) 1978 by:
Dr. Gabriele Tartar!
Agrochemical Department
Control Laboratory
CIBA-GEIGY S.p.A.
C.P. 88
1-2104? SARONNO (VA)
ITALY
Note: The amount of standard and sample and some dilution factors have been
changed to allow more significant figures in the calculations and to
reduce errors in weighings and making dilutions. The final
concentrations are as in the method as received.
Any criticisms, suggestions, or data concerning the use of this method will be
apprec iated.
-------
-------
TLC Identification TSD-1
*
TLC Systems for Identification of Pesticides
To facilitate the process of pesticide identification, laboratory-
prepared aluminum oxide and silica gel TLC plates were spotted with
pesticide standards, developed in a series of mobile solvents, the
spots visualized, and the Rp values recorded in tables (TLC Systems
1 and 2). The objective of this work was to speed up the identification
of a suspect pesticide by means of a rapid screening technique which
would eliminate unlikely candidates, while allowing the selection of
likely ones for further study. The suspect pesticide sample is sub-
jected to the same TLC systems as for the pesticide standards, and the
elimination-selection process proceeds after comparison of Rp values
of the unknown with those of the standards previously obtained. Owing
to changes in Rp values resulting from change in humidity, temperature,
layer thickness, pesticide purity, etc., some discretion must be used
in the selection process. It is advisable to spot several known pesti-
cides (preferably technical materials) along with the unknown to enable
compensation for these variables. For example, if the Rp values of the
known are elevated from the recorded data, the unknown spots may be
similarly elevated (this is somewhat empirical because there is no
assurance that the Rp values of different pesticides will change to the
same degree). The change in R-, resulting from change in mobile solvent
is a better criterion for the selection process than is dependence on
absolute Rp values for a given solvent system.
The data are presented, therefore, only as a general guide, vith
emphasis on the need for additional solvent systems and closer control
of variable conditions affecting spot movement. Pesticides which do
not exhibit movement in the mobile solvents listed require different
layers and/or more polar solvents. TLC is used only for initial
identification and semi-quantitation with subsequent confirmation
required by at least one other means (GLC, GC-MS, etc.).
Preparation of TLC Plates
TLC System 1 (organochlorine pesticides) - Forty grams Aluminum
Oxide G Type E (EM Laboratories, Inc., 500 Exec Blvd., Elmsford, N.Y.
10523) is slurried with 75 ml of a methanol solution containing
130 mg of silver nitrate. This will coat five 8 x 8" plates or twenty
2 x 8" plates using a DeSaga applicator set for a .38 mm layer. Plates
are air-dried about 5 minutes, dried in a 100°C oven for about 1/2 hour,
Developed by B. M. Olive, CBIB, Residue & Special Projects Unit
-------
cooled, and stored in a desiccator shielded from light. The larger
plates (accommodating 12-13 spots) were normally used for spotting
the reference pesticides (5-10 micrograms) to develop the data, and
the smaller plates for the unknown and a couple of references as a
check on Rp variation. Plates were developed to a 10 cm penciled
line, air-dried a few minutes, and exposed to unfiltered UV light
(UV sterilizer) until the spots (typically black on white background)
reach maximum intensity (usually 30-60 min.).
TLC System 2 (organophosphorus and fungicide pesticides) - Forty
grams of MN-Silica Gel G-HR/UV (distributed by Brinkman Instruments,
Inc., Cantiague Road, Westbury, N.Y. 11590) is slurried with 85 ml
distilled water and applied in a layer .38 mm thick to coat five
8 x 8" plates or twenty 2x 8" plates. Plates are air-dried until the
layer is set, then dried in a 100 -105°C oven for about 1/2 hour,
cooled and stored. The spotting and development is the same as for
the chlorinated insecticides (hexane was omitted as a mobile solvent
in TLC System 2 because few pesticides of this type move in it). After
air-drying a few minutes, plates are viewed under long and/or short
wave UV light in a UV viewing box. The location of any spots is marked
with a pencil indentation (usually spots appear dark blue on fluorescent
yellow background). The plate is next sprayed with a 2% acetone solution
of 4-(p-nitrobenzyl)-pyridine (NBP), heated at 110°C for ca 10 minutes,
and then sprayed with a 10% acetone solution of tetraethylenepentamine
(TEPA). This chromogenic treatment was developed for detection of
organophosphorus pesticides and produces blue spots on a yellowish
background (JAOAC, 47^ No. 6, 1964, p. 1094).
-------
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-------
PESTICIDE FORMULATION BIBLIOGRAPHY
This bibliography is intended to provide the pesticide formulation analyst
a fairly complete reference list of published material related to the field
of pesticides. The references have been limited to "book" or "manual"
types of sources for the sake of brevity, with no effort having been made
to include specific journal articles; however, individual journals con-
cerned with pesticides are given in Section E. Industrial methodology
and technical data material have also been excluded.
Descriptive notes for individual references were limited to those sources
dealing specifically with pesticide product analysis and some of the more
important residue and technical reference sources. Enough description of
some of the instrumental methods was given so an analyst without the
specified source could possibly complete a needed analysis, if necessary.
The information would also enable one to cross-reference certain quoted
methods with industrial or other methods that may be available in the
laboratory.
Some references are dated and are probably available only through a
library. They were included, however, for the sake of completeness—
some of them offering interesting reading, if only from a historical
standpoint.
The bibliography is by no means considered to be complete. Further
additions for any of the sections would certainly be welcome for a
later edition. Please address correspondence or comments to either:
Dean F. Hill
EPA, Region IX
Pesticide Product Laboratory
50 Fulton Street, Room 545
San Francisco, California 94102
or
Warren R. Bontoyan
EPA-Office of Pesticide Programs
Technical Services Division
Chemistry Laboratory
Room 101, Bldg. 306, ARC-East
Beltsville, Maryland 20705
-------
PESTICIDE BIBLIOGRAPHY
A. PESTICIDE FORMULATION ANALYSIS
1. Official Methods of Analysis of the Association of Official
Chemists, 12th Edition, 1975. Published by the Association of
Official Analytical Chemists, P. 0. Box 540, Benjamin Franklin
Station, Washington, I). C. Supplements issued annually.
The methods described in the A.O.A.C., have been subjected to
interlaboratory collaboration and shown to be statisti-
cally reliable. These methods are the most official methods
available from an enforcement standpoint and should be used,
if possible, to substantiate any suspected violations.
Chapter 6 deals with pesticide formulations. Unfortunately,
the scope of coverage is severely limited in terms of the
types and mixtures of pesticides being currently used, and
one must resort to other methods.
Other potentially useful methods to the pesticide formulation
chemist are:
1. Acetone (GLC, 36.011)
2. Benzocaine (Colorimetric, 38.134)
3. Dichlorophene (UV, 39.120)
4. Ethanol (GLC, 36.011)
5. Glycerol (Titrimetric, 35.075)
6. Griseofulvin (UV, 42.273)
7. Hexachlorophene (UV, 35.023)
8. Isopropanol (GLC, 36.011)
9. Nicotine (UV, 42.087)
10. Paraldehyde (GLC, 37.105)
11. Phenothiazine (GLC, 38.178)
12. Phenothiazine (Colorimetric, 42.121)
13. Propylene Glycol (GLC, 35.007)
14. Propylene Glycol (Titrimetric, 19.006)
15. Ronnel (GLC, 42.141)
16. Ronnel (UV, 42.144)
17. Sulfaquinoxaline (Colorimetric, 42.168)
18. Thiabendazole (Colorimetric, 42.180)
19. Thymol (Titrimetric, 37.143)
Other pertinent sections are Ch. 2 (Fertilizers), Ch. 3 (Plants),
Ch. 29 (Pesticide Residues), and Ch. 20 (Food Additives). Chap-
ters 49, 50, and 51 deal with Spectroscopic Methods, Standard
Solutions, and Laboratory Safety, respectively.
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2. Manual of Chemical Methods for Pesticides and Devices, Environ-
mental Protection Agency, Office of Pesticide Programs, Technical
Services Division, Chemical and Biological Investigations Branch.
Issued 1975 and will be updated as needed.
The manual contains standard and generally accepted methods for
pesticide product analysis which have not yet been subjected to
interlaboratory collaboration. The methods are usually accepted
by enforcement authorities, but should be checked by two or more
different methods, whenever possible, for suspected violative
samples. There are also sections on representative IR spectra
for many of the agricultural pesticides, techniques of analysis,
TLC procedures, NMR spectra, and cross-reference method index.
3. Analytical Methods for Pesticides, Plant Growth Regulators,and
Food Additives, Ed. Gunter Zweig, Academic Press, N. Y. Vols.
I-VII; Vols. VI and VII also edited by Joseph Sherma.
Volume I (1963), Principles, Methods and General Applications
Chapters 2 and 23 concern formulation analysis; however,
both are somewhat dated and should be read with this in mind.
Other sections of general interest are Chapter 8 (Spectropho-
tometric Methods), Chapter 11 (Total Halide Analysis), and
Chapters 15-17 on bioassay techniques. Most of the other
material in Volume I has been updated in later volumes, or
has little application to the formulation chemist.
Volume II (1964), Insecticides
Individual insecticides- are covered by review articles giving
information on names, producers,and chemical, physical, and
biological properties. In addition, methods for formulation
and residue analysis are presented, and one or several of each
type are given in detail for each insecticide discussed. The
formulation methods are primarily derived from industrial
sources, as most of the articles have been written by repre-
sentatives of the companies that manufacture the different
insecticides.
Volume III (1964), Fungicides, Nematocides and Soil Fumigants,
Rodenticides, and Food and Feed Additives
Review articles with the same format as Volume II but covering
pesticides from the classes listed in the above title.
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Volume IV (1964) , Herbicides
A continuation of the individual pesticide series but
covering herbicides only.
Volume V (1967) , Additional Principles and Methods of Analysis
Chapter 1 contains an introduction to gas chromatographic
detectors, although the material is primarily oriented
toward pesticide residue analysis. Other general chapters
of interest cover thin-layer chromatography, polarography,
and residue analysis for water, fish, and wildlife samples.
There is also an introduction to techniques used in metab-
olism studies of pesticides.
Volume VI (1972) , Gas Chromatographic Analysis
Chapter 4 pertains specifically to the application of GLC
techniques to pesticide formulation analysis. Detectors,
columns, and sample preparation techniques are discussed,
as well as the precision to be expected for different types
of peak measurements. References are also presented for the
gas chromatographic analysis of different classes of pesti-
cide compounds by liquid phase. Standard deviations to be
expected from GLC procedures as well as other typical
analytical methods are given.
Other general chapters in Volume VI pertain to residue sample
preparation, detectors, and qualitative analysis. In addition,
the different chemical classes of pesticides are covered,
primarily from the residue standpoint, such as chlorinated
hydrocarbons and organophosphates, with the remainder grouped
together in a chapter on miscellaneous compounds.
Volume VII (1974), Thin-layer and Liquid Chromatography and
Analysis of Pesticides of International Importance
4. Infra-red Analysis of Pesticide Formulations, by S. W. Goza,
Virginia Department of Agriculture and Commerce, Division of
Technical Services, Richmond, Virginia 23219
This loose-leaf volume contains many useful methods for the infra-
red analysis of agricultural pesticide formulations. Although
some of the methods are identical or similar to those given else-
where, there are many that are unique to this collection. Both
dry and liquid formulation methods are described. Different
sample preparation techniques are referred to in each method
according to the type of formulation. Many of the methods are
applicable in the presence of other co-formulated pesticides.
-------
The VDA IR methods, not listed as tentative, are well recognized
by enforcement authorities; however, alternative methods should be
used to substantiate any suspected violative samples, whenever
possible. Many of these methods will be part of "EPA Manual of
Chemical Methods for Pesticides and Devices."
5. CIPAC Handbook, Volume I, Analysis of Technical and Formulated
Pesticides. Compiled by R. D. Ashworth, J. Henriet,and J. F.
Lovett; edited by G. R. Law. Collaborative International Pesti-
cides Analytical Council Limited, 1970. Published by W. H.
Heffer and Sons Ltd., Cambridge, England.
This handbook is a compilation of assay methods and other testing
procedures for examining pesticidal technical materials and formu-
lated products. The methods are those adopted by the CIPAC, and
are used primarily in Europe. The assay methods are mainly wet
chemical and spectrophotometric; however, individual procedures
are described in detail for technical materials, dusts, granulars,
wettable powders,and emulsifiable concentrates. The methods are
classified as: "CIPAC Methods," which have been investigated
collaboratively, "CIPAC Provisional Methods," which have found
wide usage but lack collaboration, and "CIPAC Draft Methods,"
which should be considered tentative at best.
Of equal value to the assay methods described in the CIPAC Hand-
book are the miscellaneous physical, stability, and by-product
determinations that are presented. Flash point, viscosity,
moisture content, suspendibility, flowability, particle size
distribution,and wettability are covered in Chapter 7 (Miscel-
laneous Techniques) and referred to in the main text under the
individual pesticides. Solubility measurements, hydrolyzable
and total chlorine determinations, dye removal, and accelerated
storage tests are also described. Chapter 7 also covers prep-
aration and criteria for purity of chemicals and reagents used
in pesticide analysis.
Chapter 8 covers the preparation of pure pesticides for use as
analytical standards. Purification steps and purity criteria
are described for: 2,4-D, MCPA, Dieldrin, Aldrin, Endrin, Rote-
none, DNBP, Ovex, Fenson, Diquat, and Paraquat.
Among the various chemical assay procedures described, the fol-
lowing may be of use to the formulation analyst. It must be
kept in mind, however, that these methods have no official status
in the U. S. Some methods have been Jointly adopted by both CIPAC
and AOAC; however, these methods are described in the AOAC.
Official "CIPAC Methods" have been designated with an asterisk.
-------
1. ANTU (Titrlmetric) , p. 16
2. Captan (IR, 1264 cm~J7CHBr3) , p. 172*
3. Captan (IR, 11.30 cnTVCHClO , p. 174*
4. CIPC (Hydrolysis/Titration), p. 223*
5. 2,4-D Esters (Hydrolysis/Titration) , p. 249-56*
6. Dalapon-Na (Colorimetric) , p. 274*
7. Dalapon-Na (Titrimetric) , p. 276
8. Dimefox (Differential Hydrolysis), p. 329
9. DNBP (UV), p. 337*
10. Endosulfan (Chromatography/IR) , p. 361
11. Endothion (Hydrolysis/Titration), p. 373*
12. Fenson (Hydrolysis/Titration), p. 392*
13. Ferbam (UV, 410 nyi/CHC^) , p. 397*
14. Gamma BHC (Hydrolyzable Chlorine), p. 986
15. Gamma BHC (Polarography), p. 37*
16. IPC (Hydrolysis/Titration), p. 593
17. MCPA (Extraction/Titration) , p. 475*7477
18. MCPA (IR, 808 cnTVacetone) , p. 482*
19. MCPA Esters (Hydrolysis/Titration), p. 499*
20. Methyl Guthion (Colorimetric), p. 24
21. Ovex (Hydrolysis/Titration), p. 213*
22. Petroleum Oils (Gravimetric - Neutral Oil Content), p. 582
23. Rotenone (Colorimetric), p. 610
24. Schradan (Differential Hydrolysis), p. 621
25. Sodium Chlorate (Titrimetric/Iodimetric) , p. 626*
26. Sodium Chlorate (Titriraetric/K2Cr207) , p. 628*
27. Sodium Trichloroacetate (Decarboxylation/Titration) , p. 691*
28. Sulfur (Titrimetric), p. 632*
29. 2,4,5-T (Extraction/Titration), p. 642*
30. 2,4,5-T Esters (Hydrolysis/Extraction/Titration) , p. 646*& 651*
31. TCNB (Polarography), p. 663*
32. TEPP (Selective Hydrolysis/Titration), p. 667
33. Thirara (Dimethylamine Distillation), p. 677
34. Trichloroacetic Acid (Decarboxylation/Titration), p. 691*
35. Warfarin (UV, 305 imi in CHC13) , p. 698 6. 699*
The following infra-red procedures are described in general terms
in Chapter 5 (p. 730-733). The extraction procedure used for
dusts and wettable powders is diethyl ether/Biichner funnel rinsing.
These methods should be considered strictly as tentative.
1. Allyl Alcohol (920 cm~1/CS2)
2. Bromophos (715 cnTVCSo)
3. Chlorbenside (818 cm~1/CS2)
4. 2,4-D (720 cm /acetone)
5. Difolatan (1732 cn
6. Diuron (822 cm /acetone)
7. 2,4-DP (799 cm~l/CS2)
-------
8. Fenson (749 cm" /CS-)
9. Kelthane (532 cuTVCS)
10. Linuron (806 cn
11. MCPP (801 cm"1/CS2)
12. Methoxychlor (618 cnT
13. Monuron (835 cm'Vacetone)
14. Ovex (768 cm~1/CS2)
15. Pentachlorophenol (767 cm~VCS2 - in presence of
2,3,4, 6-Tetrachlorophenol )
16. Piperonyl Butoxide (940 cm~1/CS2)
17. Pyrazon (825 cm""1/DMF)
18. Ronnel (962 cnfVcSj)
19. Tetradifon (582 cm" /CS2)
20. 2, 3, 4, 6-Tetrachlorophenol (751 cm~1/CS2 - in presence of
Pentachlorophenol )
21. Thiometon (658
22. Thiram (981 or 858 cnTVCS,)
23. Toxaphene (1299 cn
6. Standard Methods of Chemical Analysis, 6th Edition, N. Ho we 11
Furman, Ed., (Volume I), Frank J. Welcher, Ed., (Volumes II and III),
D. Van Nostrand and Co., Inc., Princeton, N. J., 1962 (3 volumes)
Volume I IB, Chapter 39 specifically deals with pesticide formulation
analysis. However, most of the methods presented are wet chemical
or spectrophotometric methods available from the first three
references. The methods are quite dated, the A.O.A.C. references
being from the 1960 edition.
Other chapters of interest to the formulation chemist are:
Chapter 37 (Paint, Varnish, and Lacquer), Chapter 40 (Petroleum
and Petroleum Products) , and Chapter 45 (Soaps and Detergents) .
Volume I covers the analysis of individual elements in a variety
of chemical forms. There are several specific methods, not quoted
elsewhere, that can be useful to the pesticide product analyst,
among which are:
1. Available Chlorine (lodimetric titration) , p. 341
2. Chlorate (KBrO,/As +4+ titration), p. 343
3. Chromate (Soluble), p. 360
4. Titratable Iodine (Thiosulfate & As titration), p. 451
5. Manganese (Gravimetric), p. 643
6. Silver (with Potentiometric modification), p. 982
7. Tin (Dithiol reaction), p. 1082
In addition, there is a useful section in the back of Volume I
on the preparation of different laboratory reagents and solutions.
-------
Volume IIA covers noninserumental methods for industrial and
natural products. Sections of interest include chapters on
laboratory apparatus (p. 3), specific and selective precipi-
tants (p. 101), and the analysis of acids and bases (p. 534).
There is also an interesting chapter on titration methods
(p. 254).
Volume IIIA gives an introduction to the theory and application
of most of the common instrumental techniques of analysis.
Volume IIIB covers specific instrumental techniques for various
types of compounds, classified by usage. Topics of conceivable
interest to the pesticide formulation chemist are: Fertilizers
(p. 1102), Organic Functional Groups (p. 1162), Paints, Varnish,
and Lacquer (p. 1265), and Petroleum and Petroleum Products
(p. 1506). There is also a specific chapter on pesticide
residue analysis (p. 1464); however, it also is rather dated,
as most of the techniques have been supplanted.
7. Analysis of Insecticides and Acaricides, by F. A. Gunther and
R. C. Blinn, Interscience Publishers, Inc., New York, 1955.
This reference is quite dated, there being no material relating
to thin-layer, gas, or liquid chromatography. There is some
good background material on residue loss and decomposition after
field treatment, particularly for chlorinated hydrocarbons. Sam-
pling and sample preparation for residue analysis are covered,
but primarily for colorimetric and other methods now obsolete.
There is a section in the appendix giving UV and IR spectra for a
group of pesticides that may be of value to the formulation chemist,
although the selection is pretty much limited to chlorinated hydro-
carbons, natural products, etc.
Chapter 15 gives formulation and residue methods for quite a large
number of compounds. Most of the formulation methods, however,
are elemental in nature and have since been replaced by more
specific means of analysis. There are some methods in this source,
though, not mentioned in previous references that may be of some
use to the pesticide product analyst in certain situations. Among
these are:
1. Acrylonitrile (Cyanoethylation/Titration), p. 264
2. D-D (Bromination/Titration), p. 404
3. Ethylene Oxide (Precipitation/Titration), p. 451
4. Metaldehyde (Depolymerization/Titration), p. 479
5. Methoxychlor (Hydrolyzable chlorine), p. 347
6. Perthane (Hydrolyzable chlorine), p. 347
7. Schradan (Hydrolysis/Titration), p. 577
8. Sodium Selenate (Gravimetric), p. 582
9. TDK (Hydrolyzable chlorine), p. 347
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8. Reagent Chemicals and Standards, 5th Edition, Joseph Rosin,
D. Van Nostrand Company, Inc., Princeton, N. J., 1967
This volume contains many impurity tests and analytical assays
for common laboratory reagent chemicals to determine their
purity. There are also volumetric tables in the back that can
be very useful in determining what equivalent weight to use
for a particular titration.
Among the assay procedures, there are a number that may be of
use to the formulation chemist, particularly for technical
materials. Most of those listed below are titrimetric pro-
cedures.
1. Benzaldehyde, p. 78
2. Benzoic Acid, p. 82
3. Chloramine T, p. 129
4. Cupric Oxide, p. 157
5. Ferrous Ammonium Sulfate, p. 203
6. Ferrous Sulfate, p. 206
7. Hydrochloric Acid, p. 224
8. Hydrogen Peroxide, p. 228
9. Phosphoric Acid, p. 346
10. Potassium Bisulfate, p. 361
11. Potassium Chrornate, p. 372
12. Potassium Permanganate, p. 393
13. Potassium Persulfate, p. 394
14. Silver, precipitated, p. 427
15. Sodium Bisulfate, p. 443
16. Sodium Bisulfite, p. 445
17. Sodium Borate, p. 448
18. Sodium Carbonate, p. 452
19. Sodium Chlorate, p. 453
20. Sodium Fluoride, p. 460
21. Sodium Hydrosulfite, p. 463
22. Sodium Hydroxide, p. 464
23. Sulfuric Acid, p. 509
24. Trichloroacetic Acid, p. 531
9. American Wood-Preserver's Association Standards, Published by the
American Wood-Preserver's Association, 1012 Fourteenth Street, N.W.,
Washington, D. C. 20005 (Revised periodically)
Section A of this manual contains methods specifically applicable
to the analysis of wood preservatives, both in formulations and
treated surfaces. The methods, derived mainly from ASTM sources,
have been adopted by the AWPA as official, but their application
in pesticide enforcement situations is virtually untested, except
for those that are similar to those in previously quoted sources.
-------
Results derived from the use of these methods should be con-
firmed, whenever possible, by the use of alternate methods for
suspected violative samples. Most of the methods are wet
chemical in nature, with very little in the way of modern
instrumentation involved; thus much of the material presented
may be of historical interest only.
The following topic areas are covered:
1. Creosote (Water content, petroleum oil content,
specific gravity, etc.)
2. Waterborne Preservatives
a. Ammoniacal Copper Arsenite (NH^, As, & Cu determination)
b. Chromated Copper Arsenate (NH-j, As,& Cr determination)
c. Chromated Zinc Chloride (Cl, Zn, & Cr determination)
d. Copperized Chromated Zinc Arsenate (Cu, Cr, Zn, & As
determination)
e. Fluor Chrom Arsenate Phenol (F, DNP, PCP-Na, Cr, & As
determination)
3. Oil-borne Preservatives
a. Pentachlorophenol (Total acidity, total chlorine,
and a colorimetrie assay)
B. ADDITIONAL REFERENCE SOURCES FOR CHEMICAL INFORMATION ON PESTICIDES
AND THEIR ANALYSIS
(Later editions of some of these sources may be available.)
1. Acceptable Common Names and Chemical Names for the Ingredient
Statement on Pesticide Labels, 3rd Edition, prepared by R. L.
Caswell et al, Office of Pesticide Programs, EPA, Washington, D. C.
20460 (1975).
2. Advances in Pest Control Research. Ed. R. L. Metcalf, Interscience
Publishers, Inc., New York, 1957.
3. Agricultural Chemicals, W. R. Thomson, Thomson Publications, P. 0.
Box 989, Davis, California, 1967 revision. Issued in A volumes.
4. Applications of Nuclear Magnetic Resonance Spectroscopy in Organic
Chemistry, 2nd Ed., L.M. Jackman and S. Sternhell, Pergamon Press,
New York, 1969.
5. Atomic Absorption Spectroscopy. G. Christian and F. Feldman,
Wiley Interscience, New York, 1970.
6. Basic Gas Chromatography. H. M. McNair and E. J. Bonelli, Consol-
idated Printers, Berkeley, Calif.
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10
7. Catalog of Pesticide Standards for Pesticide Formulation
Analysis3 Environmental Protection Agency, TSD-CBIB, Beltsville, Md.
8. Chem Sources—U.S.A. 1974, Directories Publishing Company, Inc.,
Flemington, New Jersey.
9. Chemicals for Pest Control, G. S. Hartley and T. F. West,
Pergamon Press, New York, 1966.
10. The Chemistry and Action of Insecticides, H. H. Shepard,
McGraw-Hill Book Co., New York, 1951.
11. Chemistry and Mode of Action of Herbicides^ A. S. Crafts, Inter-
science Publishers, New York, 1961.
12. The Chemistry of Organophosphorus Pesticides, K. J. Schmidt and
C. Fest, Springer-Verlag, New York, 1973.
13. Chemistry of the Pesticide, Donald E. Frear, 3rd Edition, D. Van
Nostrand Co., Inc., New York, 1955.
14. Chemistry of the Pesticides, N. M. Melnikov, Edited by F. A.
Gunther and J. D. Gunther, Translated by R. L. Busbey, Springer-
Verlag, New York, 1971.
This is one of the best contemporary books available on the
overall chemistry of pesticides, even though it is basically a
Russian translation.
15. Degradation of Herbicides, P. C. Kearney and D. D. Kaufman,
Marcel Dekker, Inc., 1969, New York.
16. Detergents and Emulsifiers Annual, North American Division, Pub-
lished by McCutcheon's Division, Allured Publishing Corporation,
45 North Broad Street, Ridgewood, New Jersey.
17. Disinfection, Sterilization,and Preservation, C. A. Lawrence and
S. S. Block, Lea and Febiger, Philadelphia, 1968.
18. EPA Compendium of Registered Pesticides, Issued by the Office of
Pesticide Programs, Technical Services Division, Environmental
Protection Agency. Available from: Superintendent of Documents,
U. S. Government Printing Office (Stop No. 550-1), Washington, D. C.
20402.
Issued in 5 volumes:
Volume I - Herbicides and Plant Growth Regulators
Volume II - Fungicides and Nematocides
Volume III- Insecticides, Acaricides, Molluscicides,
and Anti-Fouling Compounds
Volume IV - Rodenticides and Mammal, Bird, and Fish Toxicants
Volume V - Disinfectants
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11
19. Farm Chemicals Handbook, Published annually by Farm Chemicals.
Available from Melster Publishing Co., 37841 Euclid Avenue,
Willoughby, Ohio, 44094.
20. Gas Chromatographic Analysis of Drugs and Pesticides, Vol. 2,
Benjamin J. Gudzinowicz, 1967, Marcel Dekker, Inc., New York.
21. Gas Liquid Chromatographv. S. V. Nogare, R. S. Juvet, Jr.,
1962, Interscience Publishers, a division of John Wiley & Sons,
New York.
22. guide to the Chemicals Used in Crop Protection. 1973, E. Y.
Spencer, University of Western Ontario, Information Canada,
Ottawa.
2 3. Guide to Stationary Phases for Gas Chromato.gr achy. 1973, Ana lab s,
Inc., North Haven, Conn.
24. Herbicide Handbook of the Weed Science Society of America. 3rd
Ed., 1974, Weed Science Society of America.
25. Herbicides, Fungicides, Formulation Chemistry. Pesticide
Chemistry, Vol. V, edited by A. S. Tahori, Gordon and Breach
Science Publishers, New York, 1972.
26. Industrial Production and Formulation of Pesticides in Developing
Countries - Volume I: General Principles and Formulation of
Pesticides. Prepared by the Industrial Development Organization,
Vienna, Austria, 1972.
27. Insecticides, Fungicides,and Weed Killers, E. Bourart, 2nd Ed.,
Revised and enlarged by T. R. Burton, D. Van Nostrand Company,
250 Fourth Avenue, New York, 1925.
28. Manual of Methods for Chemical Analysis of Water and Wastes. U. S.
Environmental Protection Agency, Office of Technology Transfer,
Washington, D. C. 20460.
29. The Merck Index, P. G. Stecher (Ed.), Published by Merck & Co., Inc.,
Rahway, New Jersey (Latest Edition).
30. Modern Practice of Liquid Chromatography, edited by J. J. Kirkland,
Wiley-Interscience, a division of John Wiley & Sons, New York, 1971.
31. National Formulary XIV, 1975, Prepared by the National Formulary
Board, Published by the American Pharmaceutical Association,
Washington, D. C. Supplements issued annually.
32. Natural Pest Control Agents, Advances in Chemistry Series 53,
American Chemical Society, Washington, D. C., 1966.
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12
33. Organic Insecticides, Their Chemistry and Mode of Action,
R. L. Metcalf, Interscience Publishers, New York, 1955.
34. Pesticide Chemicals Official Compendium, Published by the
Association of American Pesticide Control Officials, Inc.,
1966. May be available from: Control Division, Kansas State
Board of Agriculture, 1032-S State Office Building, Topeka,
Kansas, 66606.
35. Pesticide Formulations, W. Van Valkenburg, Ed., Marcel Dekker,
Inc., New York, 1973.
36. Pesticide Identification at the Residue Level, Division of
Pesticide Chemistry, ACS Symposium - May 26-27, 1970, Toronto,
Canada; Francis J. Giros, Symposium Chairman, Advances in
Chemistry Series 104, American Chemical Society, Washington,
D. C., 1971.
37. pesticide Index, 4th Ed., Donald Frear, College Science Pub-
lishers, State College, Pa., 16801, 1965.
38. Pesticide Manual, H. Martin, Ed., Issued by British Crop Pro-
tection Council, 3rd Edition (1972), Available from: Mr. A. W.
Billitt, Clacks Farm, Sorely, Orabersley, Droitwich, Worcester,
England.
Probably the best handy reference source for general information
on agricultural pesticides. Although the headings are based on
British nomenclature, there is a good index to cross-reference
into American names. A brief page description for each pesticide
includes:
Chemical, common, and trade names
Manufacturing process
Stability
Chemical and physical properties
Uses
Toxicity
Types of formulations
References to formulation and residue analytical methods
39. Pyrethrum; The Natural Insecticides, J. E. Casida, Academic Press,
Inc., New York, 1974.
40. Quantitative Organic Analysis via Functional Groups, 3rd Ed.,
Sidney Siggia, John Wiley & Sons, New York, 1963.
41. Residue Reviews, F. A. Gunther, Residues of Pesticides and Other
Chemicals in Foods and Feeds, Springer-Verlag, New York.
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13
42. The Sadtler Commercial Infra-red Spectra - Agricultural Chemicals,
Available from the Sadtler Research Laboratories, 3314-20 Spring
Garden St., Philadelphia, Pa., 19104.
Includes KBr, neat,and mull spectra of acaricides, bactericides,
defoliants, fungicides, herbicides, insecticides, nematocides,
repellants and attractants, rodenticides, and miscellaneous
pesticides.
43. The Sadtler Guide to NMR Spectra, W. W. Simons, M. Zanger, Sadtler
Research Laboratories, Inc., Philadelphia, Pa.
44. Specifications for Pesticides, 2nd Ed., World Health Organization,
Geneva, Switzerland, 1961.
45. Spectrometric Identification of Organic Compounds, 3rd Ed., R.
Silverstein and G. Bassler, John Wiley & Sons, New York, 1975.
46. Spot Tests in Organic Analysis, Fritz Feigl, in collaboration
with Vinzenz Anger, translated by Ralph E. Oesper, 7th Ed.,
Elsevier Publishing Co., New York, 1966.
47. The United States Dispensatory, 24th Ed., A. Osol, R. Pratt, and
G. Farrar, Jr., J. B. Lippincott Co., Philadelphia and Toronto.
48. The United States Pharmacopeia, U.S.P. XIX (1975) By Authority
of the U. S. Pharmacopeial Convention, Inc., 12601 Twinbrook
Parkway, Rockville, Md. 20852. Supplements issued.
49. Thin Layer Chromatography, edited by Egon Stahl, 1965, Springer-
Verlag, New York.
C. JOURNALS USEFUL TO THE PESTICIDE FORMULATION ANALYST
Only the Journal of the Association of Official Analytical Chemists
publishes articles concerning pesticide formulation analysis on a
sustained basis. The other journals listed have occasional articles
on pesticide product analysis, but are generally more oriented toward
residue analysis, photo- and metabolic decomposition, toxicity, or
general analytical chemistry.
1. The Analyst, Published monthly by the Society for Analytical
Chemistry, 9/10 Savile Row, London, WlX 1AF.
2. Analytical Abstracts, Published monthly by the Society for
Analytical Chemistry, 9/10 Savile Row, London. Printed by
Heffers Printers, Ltd., Cambridge, England.
3. Analytical Chemistry,, Published monthly by the American Chemical
Society, 1155 16th St., N. W., Washington, D. C. 20036.
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14
4. Bulletin of Environmental Contamination and Toxicology, Published
monthly by Springer-Verlag, Inc., 175 Fifth Avenue, New York,
N. Y. 10010.
5. Chemical and Engineering News, Published weekly by the American
Chemical Society, 1155 16th St., N. W., Washington, D. C. 20036.
6. Environment, Published monthly by the Scientists' Institute for
Public Information, 438 N. Skinker Blvd., St. Louis, Missouri 63130.
7. Environmental Science and Technology, Published monthly by the
American Chemical Society, 1155 16th St., N. W., Washington, D. C.
20036.
8. Journal of Agricultural and Food Chemistry, Published bimonthly by
American Chemical Society, 1155 16th St., N. W., Washington, D. C.
20036.
9. Journal of the Association of Official Analytical Chemists,
Published bimonthly by the Association of Official Analytical
Chemists, Inc., Box 540, Benjamin Franklin Station, Washington,
D. C. 20044.
10. Journal of Chromatographic Science (Formerly Journal of Gas
Chromatography), Published monthly by Preston Technical Abstracts
Company, P. 0. Box 312, Niles, Illinois, 60648.
11. Journal of Chromatography, Published fortnightly by Elsevier
Publishing Company, Amsterdam, Netherlands.
12. Pesticide Abstracts (Formerly Health Aspects of Pesticides Abstract
Bulletin), Published monthly by the Environmental Protection Agency,
Office of Pesticides Programs, Technical Services Division, Rm. EB-49,
401 M Street, S. W., Washington, D. C. 20460.
13. Pesticide Chemical News, Published weekly by Louis Rothschild, Jr.,
420 Colorado Building, 1341 G Street, N. W., Washington, D. C. 20005.
14. Pesticides Monitoring Journal, Published quarterly under the auspices
of the Federal Working Group on Pest Management by the U. S. Environ-
mental Protection Agency, Office of Pesticides Programs, Technical
Services Division, Room B49, East Waterside Mall, 401 M Street, S. W. ,
Washington, D. C. 20460.
15. Science, Published weekly by the American Association for the
Advancement of Science, 1515 Massachusetts Avenue, N. W., Washington,
D. C. 20005.
-------
15
D. PESTICIDE RESIDUE ANALYSIS
Methods of multi-residue and specific pesticide residue analysis
in different media are given in references quoted in Section I as
noted. The following sources may also be of value to the pesticide
residue analyst:
1. Pesticide Analytical Manual, U. S. Department of Health,
Education and Welfare, Food and Drug Administration, 2nd
Edition (1968), revised periodically. Issued in two volumes.
Volume I - Methods Which Detect Multiple Residues
Organochlorine (both ionic and nonionic) and
organophosphate pesticide extraction procedures
and clean-ups are described. GLC, TLC, PC, and
confirmatory tests are given treatment with
respect to multi-residue analysis.
Volume II- Methods of Individual Pesticide Residues
These methods are primarily those derived from
commodity tolerance applications and petitions,
although others are also included. The methods
may or may not have had collaborative testing, but
most are referenced to published or available
literature.
2. Analysis of Pesticide Residues in Human and Environmental Samples,
Ed. J. F. Thompson; Prepared by The Primate & Pesticides Effect
Laboratory, Perrine, Florida (now located at Research Triangle
Park, North Carolina). Revised November 1972 and December 1974.
This valuable manual contains general information on sampling,
laboratory procedures, gas chromatography, and confirmatory
procedures. Chlorinated hydrocarbon and organophosphate pesti-
cide analyses in human tissue and excreta are covered, along
with urine analysis for some of the ionic pesticides. Air,
water, soil, and dust procedures are given for pesticide analysis.
PCB analysis is covered, including typical chromatograms of
different Aroclors. Mercury analysis in water, blood, urine,
and fish samples is presented along with the specific analysis
for methyl mercury.
3. Methods for Organic Pesticides in Water and Wastewater, Environ-
mental Protection Agency, National Environmental Research Center,
Cincinnati, Ohio, 1971. Revisions and additions issued periodically.
Primarily covers laboratory practices and analytical methodology
for analysis of chlorinated hydrocarbons in water and waste-
water .
-------
16
4. Guide to the Analysis of Pesticide Residues, Prepared by H. P.
Burchfield and Donald W. Johnson for U. S. Department of
Health, Education, and Welfare, Public Health Service, Bureau
of State Services (Environmental Health), Office of Pesticides,
Washington, D. C., under contract with Southwest Research
Institute, San Antonio, Texas 78206. Available from the Super-
intendent of Documents, U. S. Government Printing Office,
Washington, D. C. 20402 (issued in 2 volumes)
Volume I - Contains a compilation of methods which are recom-
mended for the analysis of pesticide residues in
water, soil, plant tissues, animal tissues, body
fluids, dairy products,and related environments.
General principles, extraction, clean-up, and gas
chromatography are covered for a variety of classes
of compounds.
Volume II- Covers non-chromatographic techniques, infra-red
identification,and a compilation of chemical and
physical properties for a number of individual
pesticides. There are a number of infra-red spectra
for different compounds, most being KBr disks, however.
5. Analysis of Organic Pollutants in Water and Wastewater, W. Leithe,
Ann Arbor Science Publishers, Inc., Ann Arbor, Michigan,1973
E. SOURCES ON PESTICIDE USAGE, TOXICITY,AND CONTROVERSY
1. Agricultural Applications of Petroleum Products, Advances in
Chemistry Series No. 7, Published by the American Chemical
Society, Washington, D. C., 1952.
2. Chlorodioxins - Origin and Fate, Advances in Chemistry Series
No. 120, Published by the American Chemical Society, Washington,
D. C., 1973.
3. Organic Pesticides in the Environment, Advances in Chemistry
Series No. 60, Published by the American Chemical Society,
Washington, D. C., 1966.
4. Pesticidal Formulations Research, Advances in Chemistry Series
No. 86, Published by the American Chemical Society, Washington,
D. C., 1969.
5. Pesticides in Tropical Agriculture, Advances in Chemistry
Series No. 13, Published by the American Chemical Society,
Washington, D. C., 1955.
-------
17
6. Scientific Aspects of Pest Control, Symposium conducted by the
National Academy of Sciences, National Research Council,
Washington, D. C., Feb. 1-3, 1966 (Publication 1402 NAS/NRC).
7. Beatty, R. G., The DDT Myth; Triumph of the Amateurs, John Day
Co., New York, 1973.
8. Beirne, B., Pest Management, CRC Press, Cleveland, Ohio, 1966.
9. Bicknell, F., Chemicals in Your Food and in Farm Produce; Their
Harmful Effects, Emerson Books, Inc., 1961.
10. Brown, A. W. A., Insect Control by Chemicals, John Wiley and Sons,
New York, 1951.
11. Burgess, H. D. and Hussey, N. W., Microbial Control of Insects,
Academic Press, New York, 1971.
12. Busvine, J. P., A Critical Review of the Techniques for Testing
Insecticides, Commonwealth Agricultural Bureau, 1971.
13. Carson, Rachel, Silent Spring. Houghton Mifflin Co., Boston, 1962.
(Available in paperback)
14. deOng, E. R., Insect, Fungus.and Weed Control, Chemical Publishing
Co., New York, 1953.
15. Dethier, V. G., Chemical Insect Attractants and Repellents,
Blakiston Co., Philadelphia, 1947.
16. Edwards, C., Persistent Pesticides in the Environment, CRC Press,
Cleveland, Ohio, 1970.
17. Epstein, S. S. and Legator, M. S., The Mutagenicity of Pesticides,
Concepts and Evaluations, MIT Press, Cambridge, Mass., 1971.
18. Gerard, H. W. and Deichmann, U. B., Toxicology of Drugs and Chemicals,
Academic Press, New York, 1969.
19. Graham, F. Jr., Since Silent Spring, Houghton Mifflin Co., Boston,
1970. (Available in paperback)
20. Gunther, F. A. and Jeppson, L. R., Modern Insecticides and World
Food Production, John Wiley and Sons, New York, 1960.
21. Headley, J. C. and Lewis, J. N., The Pesticide Problem; An Economic
Approach to Public Policy, Resources for the Future, Inc., Wash-
ington, D. C., Distributed by Johns Hopkins Press, Baltimore,
Maryland, 1967.
-------
18
22. Henkin, H. et al, The Environment, the Establishment, and the Law,
Houghton Mifflin Co., Boston, 1971.
23. Herber, L. , Our Synthetic Environment, Alfred E. Knopf, New York,
1962.
24. Hunter, B. T., Gardening without Poisons, 2nd Ed., Houghton Mifflin
Co., Boston, 1971.
25. Jacobson, M., Naturally Occurring Insecticides, Marcel Dekker,
New York, 1971.
26. Kilgore, W. and Doutt, R., Pest Control; Biological, Physical, and
Selected Chemical Methods. Academic Press, New York, 1967.
27. Lehman, A. J., Summaries of Pesticide Toxicity, Published by the
Association of Food and Drug Officials, P. 0. Box 4267, Springfield,
Illinois 62708, 1965.
28. Mallis, A., Handbook of Pest Control, 5th Ed., MacNair-Dorland
Company, New York, 1969.
29. Martin, H., Scientific Principles of Crop Protection, 5th Ed.,
Edward Arnold Ltd., London, 1964.
30. Matsumura, F. et al. (Eds.), Environmental Toxicology of Pesticides,
Academic Press, 1972.
31. McMillen, W., Bugs or People, Appleton-Century, New York, 1967.
32. Meltzer, Y., Hormonal and Attractant Pesticide Technology, Noyes
Data Corporation, Park Ridge, N. J., 1971.
33. Miller, M. W. and Berg, G. G., Eds., Chemical Fallout, Charles C.
Thomas Publishers, Springfield, Illinois, 1969.
34. Montague, K. and Montague, P., Mercury, Sierra Club, San Francisco,
1971.
35. Muirhead-Thomson, R. C., Pesticides and Freshwater Fauna, Academic
Press, New York, 1971.
36. Pfadt, R. (Ed.), Fundamentals of Applied Entomology, 2nd Edition,
MacMillan Company, New York, 1971.
37. Rodale, J. I. and Staff, Our Poisoned Earth and Sky, Rodale Books,
Inc., Emmaus, Pennsylvania, 1964.
38. Rolsten, L. and McCoy, C. E., Introduction to Applied Entomology,
The Ronald Press Company, New York, 1966.
-------
19
39. Rudd, R. L., Pesticides and the Living Landscape, University of
Wisconsin Press, Madison, Wisconsin, 1964. (Available in soft
cover)
40. Sondheimer, E. and Simeone, J. B., Chemical Ecology, Academic
Press, New York, 1970.
41. Sunshine, I. (Ed.), Handbook of Analytical Toxicology, CRC
Company, Cleveland, Ohio, 1969.
42. Swann, L., Beneficial Insects, Harper and Row, New York, 1964.
43. Wellford, H., Sowing the Wind, Grossman Publishers, New York, 1972.
44. Whiteside, T., Defoliation, Ballantine/Friends of the Earth Book,
New York, 1970. (Available in paperback)
45. Whiteside, T., The Withering Rain, America's Herbicidal Folly,
E. P. Button and Co., New York, 1971.
46. Whitten, J., That We May Live, D. Van Nostrand Co., Princeton,
New Jersey, 1966.
47. Wood, D. L. et aL, Control of Insect Behavior by Natural Products,
Academic Press, New York, 1970.
-------
Infrared Spectra — Introduction
Infrared spectroscopy is one of the most definitive ways of
characterizing a chemical compound. The infrared absorption band
pattern is analogous to a "fingerprint." With very few exceptions,
positive identification of a chemical compound can be made by com-
paring the IR spectrum of the substance in question to the IR spectrum
of known pure compounds. Small differences in the spectra will dif-
ferentiate between compounds of similar structure and also will
indicate the presence of impurities.
Infrared spectroscopy is a very useful tool for the analysis of
pesticide formulations. Quite frequently, quantitative measurements
can be made without elaborate extraction procedures just by using an
absorption band at a wavelength where no interference is present. A
linearity curve made at this chosen wavelength will determine a working
range of useful concentrations. Only this short section of the IR
spectrum is needed for quantitative calculation; however, a full scan
will provide a qualitative identification.
The infrared spectra in this section were scanned on a Perkin-Elmer
Model 521 double beam spectrophotometer using KBr disks, Nujol mull cells,
and internal reflectance attachments. The samples of pesticides were
obtained directly from the manufacturers. Samples from other sources
were purified or recrystallized when necessary. Scans were made from
4000 cm to 200 cm (2.5 ji to 50 ^i) using instrument settings as follows:
attenuator speed 11, amplifier gain 5, slit program 10, scan time 32,
speed suppression 5, beam source current 0.8 amp., and filter automatic.
-------
The spectra of samples prepared using KBr disks, solutions, mulls,
or internal reflectance will differ in the shape and intensity of the
absorption bands. For this reason it is advisable for each lab to
accumulate spectra scanned on their own instruments using concentrations,
solvents or other matrices, cells or sample holders, in accordance with
their analytical needs and interest.
The names used in the following index and table and on the indi-
vidual spectra are a combination of common, trade, and accepted proper
names. They are arranged alphabetically and the same name is used
throughout. Cross reference to other names may be made using the
"Pesticide Cross Reference Index to the Methods" section under "Methods
of Analysis" or by using the "Other Names" section under each method.
The Environmental Protection Agency expresses its gratitude to
Dr. Paul A. Giang of the Analytical Chemistry Laboratory, Agricultural
Environmental Quality Institute, United States Department of Agriculture,
Beltsville, MD 20705 for his expertise in preparing these infrared
spectra. E.P.A. also wishes to thank the U.S.D.A. for permitting use
of this material.
Mention of a pesticide or a proprietary product in this manual does
not constitute a recommendation or an endorsement of this product by the
U. S. Department of Agriculture or the U. S. Environmental Protection
Agency.
-------
In the table of "Pesticide and IR Data," the following key is
used to indicate the type of pest control provided by compounds
listed in the table:
(A) Acaricide
(B) Bactericide
(D) Disinfectant
(Fum) Fumigant
(F) Fungicide
(H) Herbicide
(I) Insecticide
(IR) Insect Repellent
(M) Molluscicide
(N) Nematocide
(PGR) Plant Growth Regulator
(R) Rodenticide
(S) Synergist
(VP) Vertebrate Poison
The analytical bands in the table indicate areas where (for
single compounds) quantltation is most feasible; however, for several
compounds present in a formulation, these peaks may not necessarily
be the best. It may be necessary to choose another absorption band
where only the component of interest absorbs, or to extract the
component of interest free from any interfering compounds.
-------
Infrared Spectra — Pesticide and IR Data
Pesticide
Name
Abate
Acaralate
Acritet
Akton
alachlor
aldicarb
aldicarb
sulfone
aldicarb
sulfoxide
aldrin
Alice Ketone
allethrin
Alodan
ametryne
Amiben
Amical-48
amid ith ion
aminocarb
4-AminO"
pyridine
amitrole
Ammate
ancymidol
antimycin
Use
I
A
Fum
I
H
I,A,N
I,A,N
I,A,N
I
R,S
I
I,A
H
H
D
I,A
I
Avi-
repel .
H
H
PGR
B
% Purity
99
98
44
90
99
92
90
92
99
99
94
97
95
tech.
tech.
88
94
100
98
98
tech.
tech.
.0
.6
.0
.5
.0
.4
.0
.2
.5
.4
.8
.8
.0
gr.
gr-
.2
.2
.0
.7
.6
gr.
gr.
IR Sample
Matrix
KBr
KBr
disk
disk
Nujol mull
IRA
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
IRA
KBr
KBr
KBr
plate
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
plate
disk
disk
disk
Nujol mull
KBr
disk
Nujol mull
Analytical Bands
Wave number
(cm )
930,
1022,
2220,
1224,
1180,
1330,
1158,
1360,
1320,
1029,
910,
1196,
1224,
1214,
1068,
1114,
1084,
1436,
1426,
792,
1298,
1516,
778
760
864
720
894
988
760
515
720
696
850
846
808
774
815
574
826
1224
876
364
824
746
Microns
10
9
4
8
8
7
8
7
7
9
11
8
8
8
9
8
9
6
7
12
7
6
.74,
.78,
.52,
.16,
.46,
.51,
.64,
.35,
.55,
.72,
.00,
.36,
.16,
.24,
.36,
.96,
22
. i.4. ,
.96,
.01,
.64,
.70,
.59,
12
12
11
13
11
10
13
19
13
14
11
11
12
12
12
17
12
8
11
27
12
oo
.87
.20
.58
.94
.18
.16
.14
.42
.64
.34
.76
.82
.84
.88
.27
.50
.08
.16
.40
.48
.12
13.42
Anti-resist-
ant/DDT
tech. gr.
KBr disk
1150, 758
8.69, 13.22
-------
Pesticide
Name
ANTU
Aramite
arsenic
trioxide
Aspon
asulam
atrazine
azinphos-
ethyl
azinphos-
methyl
azinphos-
methyl
oxygen analog
Azobenzene
Azodrin
Bandane
Banol
Banomite
Banvel M
barban
barium
carbonate
Barthrin
Baygon
benef in
benomyl
bensulide
Use
R
A
R
I
H
H
I
I
I
A
I,A
H
I
A
H
H
R
I
I
H
F
H
% Purity
tech.
90
100
94
98
99
98
99
98
98
99
99
99
98
99
tech.
100
tech.
99
90
97
97
gr.
.0
.0
.3
.6
.0
.4
.9
.0
.6
.0
.9
.5
.6
.8
gr.
.0
gr.
.2
.0
.0
.7
IR Sample
Matrix
KBr
IRA
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
IRA
KBr
IRA
KBr
IRA
disk
plate
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
plate
disk
plate
disk
plate
Analytical Bands
Wave number
(cm'1)
1338,
1506,
1252,
1458,
1082,
978,
1380,
896,
896,
1446,
1262,
1260,
1096,
1126,
1176,
1160,
854,
1032,
1365,
906,
1136,
878,
764
826
1040
746
676
798
894
540
826
772
808
856
930
740
692
896
690
926
1034
710
792
684
Microns (/u)
7
6
7
6
9
10
7
11
11
6
7
7
9
8
8
8
11
9
7
11
8
11
.47,
.64,
.99,
.86,
.24,
.22,
.25,
.16,
.16,
.92,
.92,
.94,
.12,
.88,
.52,
.62,
.72,
.69,
.33,
.06,
.82,
.38,
13
12
9
13
14
12
11
18
12
12
12
11
10
13
14
11
14
10
9
14
12
14
.07
.12
.54
.48
.78
.52
.26
.50
.06
.94
.37
.68
.75
.54
.47
.18
.48
.80
.67
.08
.67
.56
-------
Pesticide
Name
bentazon
benzadox
BHC, Alpha
Isomer
BHC, Delta
Isomer
BHC, Epsilon
Isomer
BHC, Gamma
Isomer
BHC (tech.
grade)
bifenox
binapacryl
Black Copper
Oxide
BNOA
bomyl
bromacil
bromophos
bromoxynil
bromoxynil
octanoate
Bulan
butonate
butylate
Bux
cacodylic
acid
Use
H
H
I
I
I
I
I
H
I,F
F
PGR
I
H
I,A
H
H
I
I
H
I
H
% Purity
99.9
99.9
98.4
97,
98
100
tech.
98
99
100
99
92
tech.
94
96
97
96
95
99
tech.
98
.8
.8
.0
gr.
.7
.0
.0
.0
.0
gr.
.4
.4
.0
.4
.0
.5
gr.
.8
IR Sample
Matrix
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
Analytical Bands
Wave number
(cm'1)
1230,
1158,
1096,
1026,
1298,
1100,
1340,
970,
740,
1072,
916,
1074,
1338,
2260,
1546,
1404,
1762,
1224,
948,
748,
744
790
948
920
716
778
850
822
796
840
808
760
716
1578
748
682
922
724
698
650
Microns (p)
8.14,
8,
9
9
7
9
7
10
10
9
10
9
7
4
6
7
5
8
10
13
.63,
.12,
.74,
.70,
.09,
.46,
-31,
.64,
.33,
.92,
.31,
.47,
.42,
.47,
.12,
.68,
.17,
.54,
.44,
13.44
12,
10
10
13
12
11
12
12
11
12
13
13
6
13
14
10
13
14
15
.62
.56
.90
.96
.85
.76
.17
.58
.90
.38
.14
.94
.34
.44
.68
.84
.82
.32
.38
-------
Pesttcide
N.me
cadmium
chloride
captafol
captan
carbaryl
carbopheno-
thion
carboxin
CDAA
CDEC
Use
F
F
F
I
I,A
F
H
H
% Purity
99
99
99
99
92
100
98
97
.4
.6
.6
.0
.4
.0
.0
.5
IR Sample
Matrix
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
disk
disk
disk
disk
disk
disk
disk
disk
Analytical Bands
Wave number
(cm )
1878,
996,
1378,
932,
1470,
1278,
984,
1200,
816
880
770
1090
780
788
826
Microns (u)
5
10
7
10
6
7
10
8
.34
.06,
.26,
.68,
.80,
.82,
.16,
.33,
12
11
13
9
12
12
12
.26
.35
.02
.17
.82
.68
.12
cetyl pyridinium
bromide
chinothionate
Chloranil
chlorbenside
chlorbromuron
chlordane ,
alpha isomer
chlordane ,
gamma isomer
chlordane
(tech. grade)
chlordene
chlordimeform
chlorfen-
vinphos
Chlorflu-
recol
chlormequate
chloride
D
A,F
F
I, A
H
I
I
I
I
I,A
I
PGR
PGR
98
94
96
98
94
100
100
tech.
98
86
92
tech.
99
.2
.0
.8
.0
.0
.0
.0
gr.
.8
.4
.0
gr-
.8
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
disk
disk
disk
disk
disk
disk
disk
disk
disk
Nujol mull
KBr
KBr
KBr
disk
disk
disk
966,
1116,
1098,
1002,
1226,
1160,
1250,
1434,
1596,
1094,
1576,
1140,
1288,
770
758
748
748
872
826
820
748
1178
808
920
420
450
10
8
9
9
8
8
8
7
6
9
6
8
7
.36,
.96,
.11,
.98,
.16,
.62,
.00,
.03,
.26,
.14,
.34,
.77,
.76,
13
13
13
13
11
12
12
13
8
12
10
23
22
.00
.18
.37
.38
.45
.11
.18
.38
.49
.36
.84
.36
.20
-------
Pesticide
Name
chlorobenzi-
late
chloroneb
chlorophac-
inone
chlorothalo-
nil
chloroxuron
chlorpropham
Chlorthton
Ciodrin
Citronella
copper
arsenate
copper sulfate
pentahydrate
coumachlor
coumaphos
Counter
cyanamide
cycloate
cyclo-
heximide
Cyolane
cyprazine
2,4-D
Use
A
F
R
F
H
H
I
I
IR
I,F
F
R
I
I
H,F
H
F
I
H
H
% Purity
100
98
100
94
96
99
98
89
100
99
100
98
98
94
100
99
98
98
99
100
.0
.6
.0
.6
.4
.9
.9
.0
.0
.4
.0
.6
.9
.4
.0
.0
.2
.2
.6
,0
IR Sample
Matrix
KBr
KBr
KBr
KBr
KBr
KBr
KBr
disk
disk
disk
disk
disk
disk
disk
Nujol mull
IRA
IRA
IRA
KBr
KBr
KBr
KBr
KBr
plate
plate
plate
disk
disk
disk
disk
disk
Nujol mull
KBr
IRA
KBr
disk
plate
disk
Analytical Bands
Wave number
(cm-1)
1010,
1082,
1008,
976,
1300,
1278,
1346,
906,
1378,
828,
968,
1070,
1336,
1154,
2074,
1226,
1032,
1240,
888,
1300,
752
860
580
690
748
878
748
698
1226
438
652
760
1142
650
664
848
450
866
800
792
Microns (/u)
9
9
9
10
7
7
7
11
7
12
10
9
7
8
4
8
9
8
11
7
.90,
.24,
.92,
.24,
.69,
.82,
.43,
.04,
.25,
.07,
.34,
.34,
.48,
.66,
.82,
.16,
.69,
.06,
.26,
.69,
13
11
17
14
13
11
13
14
8
22
15
13
8
15
15
11
22
11
12
12
.30
.62
.24
.49
.36
.38
.36
.32
.16
.84
.34
.14
.75
.40
.10
.78
.36
.54
.50
.68
2,4-D, butoxy-
ethyl ester
2,4-D, butyl
ester
H
98.9
98.4
KBr disk
KBr disk
1196, 868
1476, 798
8.36, 11.52
6.77, 12.54
-------
Pesticide
Name Us e
2,4-D, ethyl-
hexyl ester H
2,4-D, iso-
octyl ester H
2,4-D, iso-
propyl ester H
dalapon H
dalapon-Na H
daminozide PGR
Dasanit I,N
Dasanit
(0- Analog) I,N
Dasanit
(0-Sulfone) I,N
Dasanit
Sulfone I,N
dazomet N
2,4-DB H
DCPA H
DDA I
DDE I
DDT I
p,p'-DBrDT I
Deet IR
DEF I
demeton-0,
sulfone I, A
demeton-0,
sulf oxide I, A
7o Purity
97
99
97
98
85
100
tech.
tech.
tech.
tech.
99
98
tech.
99
100
100
100
98
95
90
94
.4
.6
.0
.0
.4
.0
gr.
gr.
gr.
gr.
.4
.2
gr.
.4
.0
.0
.0
.0
.2
.0
.4
IR Sample
Matrix
KBr
KBr
KBr
IRA
IRA
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
IRA
KBr
IRA
KBr
KBr
KBr
KBr
disk
disk
disk
plate*
plate*
disk
disk
disk
disk
disk
disk
disk
disk
disk
plate
disk
plate
disk
disk
disk
disk
Analytical Bands
Wa^e number
(cm-1)
1078,
870,
800,
1254,
1442,
1012,
1210,
1480,
1142,
1102,
1220,
1022,
1398,
1086,
1380,
1096,
1078,
1160,
1200,
1324,
1246,
718
646
646
1074
872
788
532
1154
754
750
870
738
836
732
502
716
836
744
574
560
556
Microns (;j)
9
11
12
7
6
9
8
6
8
9
8
9
7
9
7
9
9
8
8
7
8
.27,
.52,
.50,
.97,
.93,
.88,
.27,
.76,
.76,
.07,
.20,
.78,
.15,
.21,
.24,
.01,
.27,
.62,
.32,
.55,
.02,
13
15
15
9
11
12
18
8
13
13
11
13
11
13
19
13
11
13
17
17
17
.92
.47
.48
.31
.46
.68
.80
.66
.26
.34
.48
.55
.94
.64
.93
.96
.97
.44
.43
.84
.96
-------
Pesticide
Name
demeton-S
sulfone
demeton-S
sulfoxide
demeton,
tech. grade
demeton (thiol
isomer)
demeton (thiono
isomer)
desmedipham
Dexon
diallate
diazinon
diazoben
dibromochloro-
propane
3,5-dibromo-
salicylanilide
4,5-dibromo-
salicylanilide
dibutalin
dicamba
dicapthon
dichlobenil
dichlone
dichloran
Dichlofen-
thion
p-dichloro-
benzene
Use
I, A
I, A
I,A
I,A
I,A
H
F
H
I
F
N
F
F
H
H
I
H
F
F
I,N
Fum
% Purity
92.
88.
tech.
91.
94.
99.
98.
99.
96.
96.
tech.
100.
99.
98.
99.
99.
97.
99.
92.
96.
100.
6
6
gr.
6
0
0
0
0
2
8
gr.
0
8
6
6
8
8
2
0
4
,0
IR Sample
Matrix
KBr
IRA
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
disk
plate
disk
disk
disk
disk
disk
disk
disk
disk
Nujol mull
KBr
KBr
disk
disk
Nujol mull
KBr
KBr
KBr
KBr
KBr
KBr
IRA
disk
disk
disk
disk
disk
disk
plate*
Analytical Bands
Wave number
(cm-1)
1308,
886,
1136,
1250,
1162,
1060,
1160,
1034,
1584,
1160,
962,
1162,
1000,
1184,
1176,
1296,
1198,
1136,
1148,
1160,
1880,
1136
610
600
620
824
688
718
822
1154
718
496
752
500
760
690
724
722
714
898
560
1016
Microns (p)
7
11
8
8
8
9
8
9
6
8
10
8
10
8
8
7
8
8
8
8
5
.10,
.28,
.80,
.00,
.61,
.43,
.62,
.67,
.31,
.62,
.38,
.60,
.00,
.44,
.50,
.72,
• 35,
.80,
.71,
.62,
.32,
8
16
16
16
12
14
13
12
8
13
20
13
20
13
16
13
.80
.40
.44
.14
.14
.54
.92
.14
.66
.92
.14
.30
.00
.14
.94
.84
13.82
14
11
17
9
.02
.16
.84
.84
-------
Pesticide
Name
dichlorprop
dichlorvos
Dicoumarol
dicrotophos
dieldrin
Dilan
dime fox
dimethoate
dimethoate,
oxygen analog
dimethyl
phthalate
dimetilan
dinitramine
dinobuton
dinoseb
dioxacarb
dioxathion
dioxathion
diphacinone
diphenamid
Diphenatrile
diphenyl
diphenylamine
Dipropalin
diquat
dibromide
Use
H
I
R
I
I
I
I, A
I
I
IR
I
H
A,F
H
I
I
I
R
H
H
F
I
H
H
7o Purity
98
99
97
97
99
tech.
98
99
92
94
99
94
99
99
tech.
tech.
94
98
100
99
97
99
98
99
.8
.6
.4
.2
.2
gr.
.6
.3
.6
.6
.4
.6
.4
.0
gr.
gr.
.0
.8
.0
.9
.9
.9
.6
.9
IR Sample
Matrix
KBr
IRA
KBr
KBr
KBr
KBr
KBr
KBr
KBr
disk
plate*
disk
disk
disk
disk
disk
disk
disk
Nujol mull
KBr
KBr
KBr
KBr
IRA
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
disk
disk
disk
disk
plate
disk
disk
disk
disk
disk
disk
disk
disk
disk
Analytical Bands
Wave number
(cm'1)
1056,
1276,
1104,
1280,
1368,
1008,
1306,
1224,
1250,
1070,
1262,
1196,
1138,
1250,
1212,
864,
956,
1142,
1140,
1076,
1340,
1170,
976,
1340,
796
848
1100
920
480
748
834
496
836
740
750
722
766
1068
750
650
822
800
748
556
802
874
766
706
Microns Qu)
9
7
9
7
7
9
7
8
8
9
7
8
8
8
8
11
10
8
8
9
7
8
10
7
.47,
.84,
.06,
.81,
• 31,
• 92,
.66,
.17,
.00,
.34,
.92,
.36,
.79,
.00,
.25,
.57,
.46,
.75,
.77,
.29,
.46,
.55,
.27,
.46,
12
11
9
10
20
13
11
20
11
13
13
13
13
9
13
15
12
12
13
17
12
11
13
14
.58
.78
.09
.86
.08
.37
.98
.18
.97
.52
.32
.83
.05
,36
.34
.38
.17
.50
,36
.98
.47
.44
.04
.16
-------
Pesticide
Name
disulfoton
dithianon
diuron
DN-111
DNBP
DNOC
dodine
Dow ET-15
DSMA
Dursban
Dyfonate
Dyrene
endosulfan
endothall
endothion
end r in
EPN
Eptam
erbon
ethephon
ethion
ethohexadiol
ethoxyquin
ethyl
dimethoate
ethyl formate
Use
I
I
H
A, I
I,H
I,H,F
F
I
H
I
I
F
I
H
I, A
I
I, A
H
H
PGR
I, A
IR
F
I
Fura
% Purity
96
97
100
98
98
99
100
98
100
99
99
98
99
99
tech.
99
98
97
tech.
98
100
tech.
100
99
96
.8
.4
.0
.0
.4
.0
.0
.6
.0
.0
.3
.4
.4
.2
gr.
.6
.8
.0
gr.
.0
.0
gr.
.0
.0
.0
IR Sample
Matrix
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
IRA
KBr
IRA
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
disk
disk
disk
disk
disk
disk
disk
disk
plate
disk
plate
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
Nujol mull
KBr
KBr
KBr
disk
disk
cell
Analytical Bands
Wave number
(cm-1)
790,
1154,
816,
1194,
1615,
1010,
1364,
1124,
—
1156,
942,
1036,
1598,
1062,
1062,
1180,
1340,
1224,
1346,
1308,
1378,
1374,
1148,
910,
990,
656
698
524
842
1256
926
706
878
—
677
624
790
750
788
746
804
686
716
870
1148
968
800
494
740
Microns (u)
12
8
12
8
6
9
7
8
8
10
9
6
9
9
8
7
8
7
7
7
7
8
11
.64,
.66,
.26,
.37,
.19,
.90,
.33,
.89,
—
.65,
.64,
.65,
.26,
.41,
.41,
.47,
.46,
.17,
.43,
.64
.26,
.33,
.71,
.00,
10.10,
15.
14.
19.
11.
7.
10.
14.
11.
-
14.
16.
12.
13.
12.
13.
12.
14.
13.
11.
8.
10.
12.
20.
24
34
12
87
96
78
16
42
-
78
02
63
34
68
42
46
57
96
48
71
33
50
16
13.52
-------
10
Pesticide
Name
ethyl
hexanediol
ethyl
trichlorfon
famphur
fenac
fenitrothion
fenson
fenthion
0-analog
fenthion
sulfone
fenthion
sulfoxide
fenthion
(tech. gr.)
fentin
hydroxide
fenuron
fenuron
ferbam
Ficam
f luometuron
fluorodif en
folpet
formetanate
Fumarin
Furadan
Fur ad an
(-3-Keto)
Use
IR
I
I
H
I,A
A
I
I
I
I
F
H
H
F
I
H
H
F
I, A
R
I,N
I,N
% Purity
tech.
94
99
98
97
99
tech.
tech.
tech.
tech.
94
99
90
98
97
98
98
99
95
tech.
98
98
gr.
.8
.2
.0
.8
.0
gr.
gr.
gr.
gr.
.6
.4
.0
.5
.0
.8
.0
.4
.0
gr.
.4
.0
IR Sample
Matrix
IRA
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
plate
disk
disk
disk
disk
disk
disk
disk
disk
disk
Nujol mull
KBr
disk
Nujol mull
KBr
KBr
KBr
KBr
KBr
KBr
disk
disk
disk
disk
disk
disk
Nujol mull
KBr
KBr
disk
disk
Analytical Bands
Wave number
(cm'1)
1456,
1152,
1232,
1338,
1340,
1010,
1154,
1228,
1220,
1220,
1316,
1300,
872,
1386,
1154,
1329,
906,
866,
1084,
872,
1336,
1024,
1370
550
704
1094
754
496
794
768
720
960
748
686
746
976
930
790
748
526
918
746
1058
758
Microns (p)
6
8
8
7
7
9
8
8
9
8
7
7
11
7
8
7
11
11
9
11
7
9
.87,
.68,
.12,
.47,
.46,
.90,
.66,
.14,
.20,
.20,
.60,
.69,
.46,
.22,
.66,
.53,
.04,
.55,
oo
• " >
.46,
.48,
.76,
7
18
14
9
13
20
12
13
13
10
13
14
13
10
10
12
13
19
10
13
13
13
.30
.14
.22
.14
.26
.17
.58
.02
.89
.45
.36
.56
.40
.24
.71
.66
.36
.04
.88
.40
.02
.19
-------
11
Pesticide
Name
Furad an
(-3-OH)
Gardona
Genite
gibberellic
acid
Glytac
Gophacide
heptachlor
heptachlor
epoxide
xachloro-
acetone
exachloro-
cyclopenta-
diene
Hormodin
Imidan
Indalone
ioxynil
ioxynil
octanoate
IPX
isobanzan
isodrin
isolan
Isoval
Karathane
karbutilate
Use
I,N
I
A
PGR
H
R
I
I
H
Fum
PGR
I
IR
H
H
H
I
I
I
I,R
A,F
H
% Purity
98
99
100
97
96
tech.
99
.6
.0
.0
.6
.2
gr.
.2
96.8
95
100
99
99
89
tech.
tech.
97
99
99
98
tech.
tech.
99
.0
.0
.2
.0
.2
gr.
gr.
.6
.5
.0
.5
gr.
gr.
.0
IR Sample
Matrix
KBr
KBr
KBr
KBr
IRA
IRA
KBr
KBr
IRA
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
IRA
disk
disk
disk
disk
plate
plate
disk
disk
plate
disk
disk
disk
disk
disk
disk
disk
disk
disk
plate
Nujol mull
Nujol mull
KBr
disk
Analytical Bands
Wave number
(cm'1)
1126,
892,
1050,
966,
1788,
1008,
1250,
1166,
644,
1142,
1272,
906,
1076,
1246,
1529,
1000,
984,
824,
1154,
744,
952,
1184,
886
578
740
770
834
584
768
818
440
708
742
714
768
896
712
790
862
588
838
540
716
994
Microns (ji)
8
11
9
10
5
9
8
8
15
8
7
11
9
8
6
10
10
12
8
13
10
8
.86,
.12,
.52,
.35,
-59,
.92,
.00,
.58,
.56,
.76,
.86,
.04,
.29,
.02,
.54,
.00,
.16,
.14,
.66,
.44,
.47,
.45,
11
17
13
12
11
17
13
12
22
14
13
14
13
11
14
12
11
17
11
18
13
10
.26
.31
.52
.98
.97
.12
.02
.22
.60
.16
.48
.02
.04
.17
.04
.66
.61
.0
.92
.52
.96
.07
-------
12
Pesticide
Name
Kelthane
Kepone
Landrin, 2,3,5-
Isomer
Landrin, 3,4,5-
Isomer
Largon
lead arsenate
lenacil
Lethane-384
linuron
Malachite
malaoxon
malathion
maleic
hydrazide
maneb
MCPA
MCPA, Iso-
octyl Ester
MCPB
MCPP
Memmi
mercuric
chloride
mercury oxide
(yellow)
Mesurol
Use
A
I
I
I
I
I
H
I
H
F
I
I
PGR
F
H
H
H
H
F
I,F
F
I
% Purity
98
85
99
99
tech.
100
99
53
98
99
tech.
98
100
98
98
tech.
99
100
99
96
100
100
.8
.0
.8
.2
gr.
.0
.6
.0
.6
.8
gr.
.4
.0
.4
.0
gr.
.2
.0
.8
.8
.0
.0
IR Sample
Matrix
KBr
disk
Nujol mull
KBr
KBr
disk
disk
Nujol mull
IRA
KBr
KBr
KBr
IRA
plate
disk
disk
disk
plate
Nujol mull
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
IRA
KBr
disk
disk
disk
disk
disk
disk
disk
disk
disk
plate
disk
Analytical Bands
Wave number
(cm'1)
1014,
1054,
932,
968,
1016,
1096,
2180,
1178,
1048,
1258,
762,
1024,
1128,
1186,
1150,
1124,
1046,
1068,
1600,
574,
1102,
504
504
686
856
774
556
1114
880
814
826
652
820
460
796
642
806
552
824
464
864
Microns (p)
9
9
10
10
9
9
4
8
9
7
13
9
8
8
8
8
9
9
6
17
9
.86,
.48,
.72,
.32,
.84,
.12,
.59,
.^9,
.54,
• 95,
.18,
.76,
.86,
.43,
.69,
.90,
.56,
.36,
.25
.42,
.08,
19
19
14
11
12
17
8
11
12
12
15
12
21
12
15
12
18
12
21
11
.84
.84
.57
.67
.92
.98
.97
.36
.28
.11
.34
.18
.76
.57
.58
.41
.13
.14
.55
.58
-------
13
Pesticide
Name
metaldehyde
Metasystox-R
methazole
methidathion
methomyl
me.thoxychlor
methyl demeton
methyl para-
thion
Methyl Tri-
thion
metobromuron
metribuzin
MGK-264
mipafox
mirex
Mobam
molinate
Monitor
monuron
Morestan
naled
naphthalaphos
naphthalene
naphthalene
acetamide
naphthalene
acetic acid
Use
M
I, A
H
I, A
I,N
I
I, A
I
I, A
H
H
S
I
I
I
H
I
H
I.A.F
I , A , Fum
I,D
Fum
PGR
PGR
% Purity
100
tech.
100
99
99
100
94
99
92
98
99
tech.
tech.
100
96
99
98
98
99
99
100
100
96
97
.0
gr.
.0
.9
.0
.0
.6
.2
.2
.0
.2
gr.
gr.
.0
.4
.3
.8
.4
.4
.0
.0
.0
.4
.4
IR Sample
Matrix
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
IRA
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
plate
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
disk
Analytical Bands
Wave number
(cm'1)
1332,
834,
1270,
1578,
1090,
1176,
1440,
1240,
1096,
1068,
1052,
1172,
1370,
882,
944,
1152,
1200,
1010,
1174,
1284,
1082,
1200,
1380,
1502,
548
576
816
644
556
782
564
764
650
446
904
718
476
532
700
660
760
830
576
806
544
1000
774
534
Microns (,u)
7
11
7
8
9
8
6
8
9
9
9
8
7
11
9
8
8
9
8
7
9
8
7
6
.51,
.97,
.87,
.34,
.17,
.50,
.94,
.06,
.12,
.36,
.50,
.53,
.30,
.34,
.50,
.68,
.33,
.90,
.52,
.79,
.24,
.33,
.25,
.66,
18.
17.
12.
15.
17.
12.
17.
13.
15.
22.
11.
13.
21.
18.
11.
15.
13.
12.
17.
12.
18.
10.
22
36
26
53
98
76
77
14
38
41
06
92
02
78
06
18
17
06
36
42
37
00
12.92
18.72
-------
14
Pesticide
Name
naptalam
N- butyl
acetanilide
neburon
Nellite
Nemacur
norbormide
norea
N-Serve
Omite
Orthene
ovex
Oxycarboxin
parathion
PCP
Pentac
pentachloro-
benzene
Per thane
Piperalin
piperonyl
butoxide
pival
prometone
pronamide
propachlor
propanil
Use
H
IR
H
N
N
R
H
B
A
I
A
F
I
H
A
H
I
F
S
R
H
H
H
H
% Purity
98
98
96
100
97
100
98
99
92
99
tech.
100
98
38
99
99
90
99
100
98
99
99
96
99
.8
.8
.4
.0
.5
.0
.8
.0
.0
.9
gr.
.0
.5
.0
.5
.9
.0
.6
.0
.0
.2
.4
.8
.2
IR Sample
Matrix
KBr
KBr
KBr
disk
disk
disk
Nujol mull
KBr
KBr
IRA
KBr
KBr
disk
disk
plate
disk
disk
Nujol mull
KBr
KBr
KBr
IRA
KBr
KBr
KBr
KBr
IRA
KBr
KBr
KBr
KBr
KBr
disk
disk
disk
plate*
disk
disk
disk
disk
plate*
disk
disk
disk
disk
disk
Analytical Bands
Wave number
(cm-1)
1346,
1210,
1032,
1018,
804,
1482,
1636,
1140,
1506,
946,
1020,
—
1160,
1440,
1014,
1410,
1120,
1032,
1044,
1136,
1018,
1094,
1160,
1196,
700
700
504
752
538
1036
1376
700
874
556
768
—
682
978
900
760
850
752
942
704
814
660
772
844
Microns (u)
7
8
9
9
12
6
6
10
6
.43,
.26,
.69,
.82,
.44,
.75,
.11,
.28,
.64,
10.64,
9
8
6
9
7
8
9
9
8
9
9
8
8
.80,
—
.62,
.94,
.86,
.09,
.92,
.65,
.57,
.80,
.82,
.14,
.62,
.36,
14
14
19
13
18
9
7
11
11
17
13
-
14
10
11
13
11
13
10
14
12
15
12
12
.28
.27
.82
.30
.58
.65
.27
.27
.44
.92
.02
-
.66
.24
.14
.15
.76
.30
.62
.21
.27
.15
.94
.04
-------
15
Pesticide
Name
pyrethrin
concentrate
Randox-T
rotenone
Ruelene
sesamex
sesone
siduron
Simazine
Sirmate
streptomycin
sulfate
strychnine
nitrate
strychnine
sulfate
Sustar
2,4,5-T
2,4,5-T
(butoxyethyl
ester)
2,4,5-T
(butyl ester)
2,4,5-T
(isooctyl
ester)
2,4,5-T
(isopropyl
ester)
2,4,5-T
(methyl
ester)
Use
I
H
I
I,A
S
H
H
H
H
B
VP
VP
H
H
H
H
H
H
H
% Purity
40
92
100
98
tech.
99
98
99
98
98
99
98
tech.
100
98
100
95
98
98
.0
.0
.0
.6
gr.
.0
.8
.1
.6
.0
.2
.0
gr.
.0
.4
.0
.5
.8
.2
IR Sample
Matrix
IRA
KBr
KBr
KBr
IRA
KBr
KBr
KBr
KBr
KBr
KBr
KBr
IRA
KBr
KBr
KBr
KBr
KBr
KBr
plate*
disk
disk
disk
plate*
disk
disk
disk
disk
disk
disk
disk
plate
disk
disk
disk
disk
disk
disk
Analytical Bands
Wave number
(cm'1)
1104,
1096,
1304,
1356,
1182,
1448,
1442,
1298,
946,
—
1270,
1592,
1300,
1134,
870,
870,
870,
830,
862,
984
778
1090
798
1036
866
1312
798
804
—
758
764
466
764
734
734
734
770
678
Microns (u)
9
9
7
7
8
6
6
7
10
-
7
6
7
8
11
11
11
12
11
.05,
.12,
.66,
.37,
.46,
.91,
.93,
.70,
.58,
-
.87,
.28,
.69,
.81,
.49,
.49,
.49,
.03,
.60,
10
12
9
12
9
11
7
12
12
-
13
13
21
13
13
13
13
13
16
.16
.88
.17
.58
.17
.56
.62
.52
.42
-
.18
.08
.46
.12
.62
.62
.62
.01
.74
-------
16
Pesticide
Name
Tabatrex
TEPP
terbacil
terbutol
thiabenda-
zole
thiram
Torak
triallate
3,4,5-
tribromo-
salicyl-
anilide
tricamba
trichloro-
carbanilide
trifluralin
Tritac
Warfarin
Zectran
zineb
Ziram
Use
IR
I
H
H
F
F
I,A
H
F,B
H
D
H
H
R
I, A
F
F
% Purity
100
40
99
97
99
tech.
97
99
100
98
98
99
99
100
92
97
91
.0
.0
.9
.0
.5
gr.
.8
.6
.0
.8
.6
.8
.0
.0
.0
.4
.4
IR Sample
Matrix
KBr
IRA
disk
cell*
Nujol mull
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
KBr
disk
disk
disk
disk
disk
disk
disk
disk
disk
Nujol mull
KBr
IRA
KBr
KBr
disk
plate
disk
disk
Analytical Bands
Wave number
(cm-1)
1722,
—
1398,
1250,
1304,
1240,
864,
1034,
1002,
1014,
1080,
904,
992,
952,
1094,
1384,
1238,
1106
—
746
850
900
848
586
810
735
584
812
704
810
702
870
974
560
Microns (p)
5
7
8
7
8
11
9
9
9
9
11
10
11
9
7
8
.81,
—
.15,
.00,
.67,
.06,
.56,
.67,
.99,
.86,
.26,
.06,
.17,
.50,
.14,
.22,
.08,
13.
—
13.
11.
11.
11.
17.
12.
13.
17.
12.
14.
12.
14.
11.
10.
17.
34
40
76
12
79
06
34
58
12
31
19
36
26
48
37
84
* Internal Reflectance Attachment
-------
INDEX
I.R. Curve
ABATE
ACARALATE
I.R. Curve
BANDANE
BANOL
ACRITET
AKTON
ALACHLOR
ALDICARB
BANOMITE
BANVEL M
BARBAN
BARIUM CARBONATE
ALDICARB SULFONE
ALDICARB SULFOXIDE
BARTHRIN
BAYGON
ALDRIN
ALICE KETANE
BENEFIN
BENOMYL
ALLETHRIN
ALODAN
BENSULIDE
BENTAZON
AMETRYNE
AM1BEN
AMICAL-48
AMIDITHION
AMTNOCARB
4-AMINOPYRIDINE
AMITROLE
AMMATE
BENZADOX
BHC (ALPHA ISOMER)
BHC (DELTA ISOMER)
BHC (EPSILON ISOMER)
BHC (GAMMA ISOMER)
BHC (TECH. GRADE)
BIFENOX
BINAPACRYL
ANCYMIDOL
ANTIMYCIN
ANTIRESISTANT/DDT
ANTU
BLACK COPPER OXIDE
BNOA
BOMYL
BROMACIL
ARAMITE
ARSENIC TRIOXIDE
BROMOPHOS
BROMOXYNIL
ASPON
ASULAM
BROMOXYNIL OCTANOATE
BULAN
ATRAZINE
AZINPHOS-ETHYL
BUTONATE
BUTYLATE
AZINPHOS-METHYL
AZINPHOS-METHYL OXYGEN ANALOG
BUX
CACODYLIC ACID
AZOBENZENE
AZODRIN
CADMIUM CHLORIDE
CAPTAFOL
-------
I.R. Curve
I.R. Curve
CAPTAN
CARBARYL
CYCLOATE
CYCLOHEXIMIDE
CARBOPHENOTHION
CARBOXIN
CYOLANE
CYPRAZINE
CDAA
CDEC
CETYL PYRIDINIUM BROMIDE
CHINOTHIONATE
CHLORANIL
CHLORBENSIDE
CHLORBROMURON
CHLORDANE (ALPHA ISOMER)
CHLORDANE (GAMMA ISOMER)
CHLORDANE (TECH. GRADE)
CHLORDENE
CHLORDIMEFORM
2,4-D
2,4-D (BUTOXYETHYL ESTER)
2,4-D (BUTYL ESTER)
2,4-D (2-ETHYLHEXYL ESTER)
2,4-D (ISOOCTYL ESTER)
2,4-D (ISOPROPYL ESTER)
DALAPON
DALAPON-Na
DAMINOZIDE
DASANIT
DASANIT (0-ANALOG)
DASANIT (0-ANALOG SULFONE)
CHLORFENVINPHOS
CHLORFLURECOL
DASANIT SULFONE
DAZOMET
CHLORMEQUAT CHLORIDE
CHLOROBENZILATE
2,4-DB
DCPA
CHLORONEB
CHLOROPHACINONE
DDA
DDE
CHLOROTHALONIL
CHLOROXURON
CHLORPROPHAM
CHLORTHION
DDT
p,p'-DBrDT
DEET
DEF
CIODRIN
CITRONELLA
DEMETON 0-SULFONE
DEMETON 0-SULFOXIDE
COPPER ARSENATE
COPPER SULFATE PENTAHYDRATE
COUMACHLOR
COUMAPHOS
COUNTER (CL-92,100)
CYANAMIDE
DEMETON S-SULFONE
DEMETON S-SULFOXIDE
DEMETON (TECH. GRADE)
DEMETON (THIOL ISOMER)
DEMETON (THIONO ISOMER)
DESMEDIPHAM
-------
I.R. Curve
I.R. Curve
DEXON
DIALLATE
DIPHENYL
DIPHENYLAMINE
DIAZINON
DIAZOBEN
DIBROMOCHLOROPROPANE
3,5-DIBROMOSALICYLANILIDE
4', 5-- DIBROMOSALICYLANILIDE
DIBUTALIN
DIPROPALIN
DIQUAT DIBROMIDE
DISULFOTON
DITHIANON
DIURON
DN-111
DICAMBA
DICAPTHON
DNBP
DNOC
DICHLOBENIL
DICHLONE
DODINE
DOW ET-15
DICHLORAN
DICHLORFENTHION
DSMA
DURSBAN
p-DICHLOROBENZENE
DICHLORPROP
DYFONATE
DYRENE
DICHLORVOS
DICOIJMAROL
ENDOSULFAN
ENDOTHALL
DICROTOPHOS
DIELDRIN
ENDOTHION
ENDRIN
OIL AN
DIMEFOX
EPN
EPTAM
DIMETHOATE
DIMETHOATE OXYGEN ANALOG
ERBON
ETHEPHON
DIMETHYL PHTHALATE
DIMETILAN
ETHION
ETHOHEXADIOL
DINITRAMINE
DINOBUTON
ETHOXYQUIN
ETHYL DIMETHOATE
DINOSEB
DIOXACARB
ETHYL FORMATE
ETHYL HEXANEDIOL
DIOXATHION
DIPHACINONE
ETHYL TRICHLORFON
FAMPHUR
DIPHSNAMID
DIPHENATRILE
FENAC
FENITROTHION
-------
I.R. Curve
I.R. Curve
FENSON
FENTHION 0-ANALOG
FENTHION SULFONE
FENTHION SULFOXIDE
FENTHION (TECH. GRADE)
FENTIN HYDROXIDE
FENURON
FERBAM
KARATHANE
KARBUTILATE
KELTHANE
KEPONE
LANDRIN (2,3,5-ISOMER)
LANDRIN (3,4,5-ISOMER)
LARGON (TH-6040)
LEAD ARSENATE
FICAM
FLUOMETURON
LENACIL
LETHANE 384
FLUORODIFEN
FOLPET
FORMETANATE
FUMARIN
LINURON
MALACHITE
MALAOXON
MALATHION
FURADAN
FURADAN (-3-KETO)
FURADAN (-3-OH)
GARDONA
GENITE
GIBBERELLIC ACID
GLYTAC
GOPHACIDE
HEPTACHLOR
HEPTACHLOR EPOXIDE
HEXACHLOROACE TONE
HEXACHLOROCYCLOPENTADIENE
HORMODIN
IMIDAN
INDALONE
IOXYNIL
MALEIC HYDRAZIDE
MANEB
MCPA
MCPA (ISOOCTYL ESTER)
MCPB
MCPP
MEMMI
MERCURIC CHLORIDE
MERCURY OXIDE (YELLOW)
MESUROL
METALDEHYDE
METASYSTOX-R
METHAZOLE
METHIDATHION
METHOMYL
METHOXYCHLOR
IOXYNIL OCTANOATE
IPX
METHYL DEMETON
METHYL PARATHION
ISOBENZAN
ISODRIN
ISOLAN
ISOVAL
METHYL TRITHION
METOBROMURON
METRIBUZIN
MGK 264
-------
I.R. Curve
I.E. Curve
MIPAFOX
MIREX
PYRETHRUM CONG.
RANDOX T
MOBAM
MOLINATE
ROTENONE
RUELENE
MONITOR
MONURON
SESAMEX
SESONE
MORESTAN
NALED
SIDURON
SIMAZINE
NAPHTHALAPHOS
NAPHTHALENE
SIRMATE
STREPTOMYCIN SULFATE
NAPHTHALENE ACETAMIDE
NAPHTHALENE ACETIC ACID
STRYCHNINE NITRATE
STRYCHNINE SULFATE
NAPTALAM
N-BUTYL ACETANILIDE
NEBURON
NELLITE
NEMACUR
NORBORMIDE
NOREA
N-SERVE
SUSTAR
2,4,5-T
2,4,5-T (BUTOXYETHYL ESTER)
2,4,5-T (BUTYL ESTER)
2,4,5-T (ISOOCTYL ESTER)
2,4,5-T (ISOPROPYL ESTER)
2,4,5-T (METHYL ESTER)
TABATREX
OMITE
ORTHENE
TEPP
TERBACIL
OVEX
OXYCARBOXIN
TERBUTOL
THIABENDAZOLE
PARATHION
PCP
THIRAM
TORAK
PENTAC
PE NTA CHLOROBENZE NE
PERTHANE
PIPERALIN
TRIALLATE
3,4' ,5-TRIBROMOSALICYLANILIDE
TRICAMBA
TRICHLOROCARBANILIDE
PIPERONYL BUTOXIDE
PIVAL
TRIFLURALIN
TRITAC
PROMETONE
PRONAMIDE
WARFARIN
ZECTRAN
PROPACHLOR
PROPANIL
ZINEB
ZIRAM
-------
ABSORBANCE
r^ r1 P P P P
en o bo b- k
ABSORBANCE
— p p p p
b bo <> W k>
-------
1- r p p
OOi O 00 O>
ABSORBANCE
p p
'*. ks
ABSORBANCE
_ -- p p p p
In O CO
-------
OOl O OO
ABSORBANCE
p o p
6- k ro
ABSORBANCE
-------
ABSORBANCE
—> -• p p p p
In o bo b- k to
ABSORBANCt
p p
k>
O
b
-------
ABSORBANCE
,.- — p p O O
JkOln O 00 O 4k IsJ
ABSORBANCE
_• _. p p p p
jkOoi o bo b> *. K>
-------
ABSORBANCE
— pop p
b bo b- *. to
ABSORBANCE
— — p p p p
4». Bin o bo '&• t*. KO
-------
ABSORBANCE
_ p p p p
o bo b- *. ro
, ABSORBANCE
1— — p p p p
it gin o bo o- *. KJ
8
-------
, ABSORBANCE
1- — p p p p
ikOLn b co b- *. k>
ABSORBANCE
i— — p p p p
i o oo 6> *- 10
-------
1 ABSORBANCE
I;- — 0 0 0 0
4k. OCn O 00 CX k k)
I ABSORBANCE
i— — p p p p
obi b bo 6- *. io
-------
ABSORBANCE
ABSORBANCE
— — p p p o
01 o OB b> V io
-------
ABSORBANCE
p p
k>
ABSORBANCE
P P
-------
ir r-p P
. OO< O 00 O>
ABSORBANCE
p p
ro
ABSORBANCE
— — p p p p
In O 'oo b» to. io
-------
ABSORBANCE
ir" r" P P P P
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