MANUAL OF CHEMICAL METHODS
     FOR PESTICIDES AND DEVICES
   U.S. Environment
     Protection /Jgency
       OFFICE OF PESTICIDE PROGRAMS
       TECHNICAL SERVICES DIVISION
CHEMICAL AND BIOLOGICAL INVESTIGATIONS BRANCH
          Published and Distributed by:
    ASSOCIATION OF OFFICIAL ANALYTICAL CHEMISTS
              July 1976

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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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                             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




(AAPCOfs 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 4 EPA pesticide formulation chemists and 2 state chemists




recognized by AAPCO as haying 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 bestin some




cases the onlymethods 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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AAPCO-EPA Editorial Committee
Warren R. Bontoyan, Chairman
EPA, TSD
Bldg. 306, Rm 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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                        ANALYTICAL METHODS

       Pesticide Name Cross Reference Index to the Methods
Aatrex

ACC 3422

Accelerate

3-(alpha-acetonyIbenzyl) -4 -
  hydroxycoumarin

3-(alpha-acetonylfurfuryl)-4-
  hydroxycoumar in

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-aminopyridine

aminotriazble
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

Animate

ammonium methanearsonate

ammonium sulfamate

Amoxone

AMS  EPA-1

p-tert-amylphenol


anilazine

Anilazine EPA-1 (tentative)

Anilazine EPA-2 (tentative)

anofex

Ansar

Antimilace

A-AP

Aphamite

Aqua-Keen

Aquathoi

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

Balfin

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
  So. 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 & Chlorophenpls
  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
Benefit! EPA-1 & 2
bis[2-(2,4-dichlorophenoxy)
  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-methyluracil

3-(A-bromo-3-chlorophenyl)-1-
  methoxy-1-methylurea
2,4-DEP


Thirara 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

Dieloran 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-methoxy-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

l-n-butyl-3-(3,4-dichlorophenyl)-l-
  methylurea

2-sec-butyl-4,6-dinitrophenol

2-sec-butyl-4,6-dinitrophenyl-3-
  methyl-2-butenoate

N-butyl-N-ethyl-or,a,a-trifluoro-
  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-
  car bamate
Metobromuron 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
                b
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




Chetnox




Chipco Turf Herbicide D
Chloroxuron EPA-1 & 2




Fluometuron EPA-1




Metobromuron EPA-1, 2, & 3




Arsenic Compounds EPA-3 & 4




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




IR




UV




HPLC




Malathion EPA-1 & 2




IR




IR




Azinphos-methyl 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-
  (raethoxymethyl)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 EPA-1


Chloropjienoxy Herbicides EPA-2

Chlorophenoxy Herbicides EPA-3
                    (tentative)

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

A-chloro-2-phenylphenol


6-chloro-2-phenylphenoI


Chlorothalonil EPA-1
Chlorothiepin

Chloro-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 potentiometric titration

Chloroxuron EPA-1 & 2

2,4-D

IR

GLC-TCD-IS

4-chloro-3,5-xylenol

Phenols & Chlorophenols
  EPA-1, 3, & 7

Fluometuron EPA-1

Pyrethrin 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)

HI

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 (tentative)

Diazinon EPA-3

Diazinon EPA-4

Dibromochloropropane EPA-1

Dibromochloropropane EPA-2 (tentative)

1,2-dibromo-3-chloropropane

4',5-dibromosalicylanilides

Dibutyl Succinate EPA-1

Dichlobenil EPA-1

Dichlone EPA-1

dichlorfenidim

p-Dichlorobenzene EPA-1 (tentative)

p-Dichlorobenzene 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

IR

GLC-FID-IS

IR

GLG-TCD

Dibromochloropropane EPA-1 & 2

Brominated Salicylanilides EPA-1

saponif ication & titration
Diuron EPA-1, 2, 3, & 4

IR

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-dlchloropropionic acid

dichlorprop


Dlcloran EPA-1

Dlcophane

Didimac

diethion

0,0-d iethyl-0-(3-chloro-A-methy1-
  2-oxo-2H-l-benzopyran-7-yl)
  phosphorothloate

0,0-diethyl S [2-(ethylthlo)ethyl]
  phosphorodithioate

0,0-diethyl S-(ethylthiomethyl)
  phosphorodithioate

0,0-diethyl 0-(2-isopropyl-6-methyl-
  A-pyrimidinyl)phosphorothioate

0,0-diethyl-O-p-nitrophenyl
  phosphorothioate

N,N-diethyl-m-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-dihydro-pyridazinedione

diisopropyl S-(2-phenylsulfonyl-
  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

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S-(Q,O-diisopropyl phosphorodithioate)
  ester of N-(2-mercaptoethyl
  benzenesulfonamide

N-2-(Q), 0-diisopropyl-phosphorothiolo-
  thionyl)ethyl benzenesulfonamide

0, 0-dimethyl dlthiophosphate of
  diethyl mercaptosuccinate

0, 0-dimethyl-0-(2-methylcarbamoyl-
  1 - methylvinyl)-phosphate

dimethyl-l-methyl-2-methyl-carbamoyl-
  yinyl phosphate

0, O-dimethyl o-p-nitrophenol
  phosphorothioate
0, 0-dimethyl S-(4-oxo-l,2,3-benzo-
  triazin-3(4H)-ylraethyl)
  phosphorodithioate

cis-3-(dimethoxyphosphinyloxy)-N-
  methylcrotonamide

dimethyl parathion
dimethyl phosphate of 3-hydroxy-N-
  methyl-cis-crotonamide

l,l-dimethyl-3-(a,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

Dinocap_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-(diphenylacetyl)-l,3-indandione




Direz




disodium methanearsonate




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

Endothall EPA-2  (tentative)
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

GLC-FID

-------
                                17
ephirsulfonate

Eptam

EPIC  EPA-1 (tentative)

EPIC  EPA-2 (tentative)

EPTC  EPA-3

EPIC  EPA-A (tentative)

EPTC  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 EP_A-_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-hexariediol




ethylhexylene glycol




Ethyl Parathlon




Etilon




Etrolene




Eurex






Fall




Falone




FBHC




Fence Rider  




fenchlorphos (ISO and BSI)




Fermide




Femes t a




Fernimine




Fernoxone




ferroprop




Ferxone




flour sulfur




flowers of sulfur




Fluometuron EPA-1




FMC 5273




FMC 5A62




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

fumarin (Great Britain,  New Zealand)

Furaazone

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
GarIon

Gearphos


Gebutox

Genitox

Gesafram

Gesapon

Gesaprim

Gesarex

Gesarol

Gesatop

Gramevin

Guesarol

Gusathlon

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 Parathion
  EPA-1, 2, 3, 4, & 5

Dinoseb EPA-1 & 2

DDT EPA-1

Prometone EPA-1 & 2

DDT EPA-1

Atrazine EPA-1 & 2

DDT EPA-1

DDT EPA-1

Siraazine EPA-1

Dalapon EPA-1

DDT EPA-1

Azinphos-methyl EPA-1

Azinphos-methyl EPA-1

DDT EPA-1


Pyrazon EPA-1

Dichlobenil EPA-1

Methiocarb EPA-1

Rotenone EPA-1

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

Hormodin

Hormotuho

Hydout

o-hydrod ipheny1

Hydrothol

6-hydroxy-3-(2H)-pyridaz inone

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

Araitrole 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


       x
Norbormide EPA-1

Bromacil EPA-1

-------
                                22
Indolebutyric acid EPA-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




Kilsera




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




IR




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




EPIC  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




Linuron EPA-1 (tentative)




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

mercaptod imethur

mercaptothion

Mercuram

Merpan

Mesurol

Meta

metacetaldehyde

Metacide
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

Metaldehyde EPA-2 (tentative)

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% arid related
  compounds 12%

methyl-1-(butylcarbamoyl)-2-
  benzimidazolecarbamate

0-methyl 0-2-chloro-4-tert-butyl-
  phenol N-methylamidophosphate

4-(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

iodimetric 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-A

Methyl Parathion EPA-5

4-(methylthio)-3,5-xylyl
  N-methylcarbamate

Metiltriazotion

metmercapturon

Metobromuron EPA-1 (tentative)

Metobromuron EPA-2 (tentative)

Metobromuron EPA-3 (tentative)

Metron


MH  EPA-1

MH-30

Mildex

2M-4Kh-M

MLT

Mocap

raonoannnonium 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-TCP-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 & titration

UV

IR

-------
                                27
Morocide




Mous-con




Mouse-tox




Moxie




MSMA




Muscatox






Nankor




1-naphthyl methylcarbamate




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




Couraaphos 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


Noraersan

Nor-Am

Norbormide EPA-1

No rex

NRDC 104

Nucidol

Nuvacron


Off

Oil of Lemongrass EPA-1 (tentative)

oil of verbena  (Indian)

Omite

Organophosphorus compounds

Organotin 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, A, & 5

Thiram EPA-1 & 2

Chloroxuron EPA-1 & 2

UV

Chloroxuron EPA-1 & 2

Resmethrin EPA-1, 2, 3, A, & 5

Diazinon EPA-1, 2, 3, & A

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

Dibromochloropropane EPA-1 & 2

Ovex EPA-1

IR

Ovex EPA-1

Ovex EPA-1

-------
                                29
Outrack

7-oxabicyclo(2,2,l)heptane-
  2,3-dicarboxylic acid
PGA

PCP

PDB

PDQ

Panthion

Paracide

paradichlorobenzene

Paradow

Paramar

Paraphos

Parathene

Parathipn 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)

I^b^il 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 & Chlorophenols EPA-1


Phenols & Chlorophenols EPA-2

Phenols & Chlorophenols EPA-3

Phenols & Chlorophenols EPA-A

Phenols & Chlorophenols EPA-5
                  (tentative)

Phenols & Chlorophenols EPA-6
                  (tentative)

Phenols & Chlorophenols EPA-7
                  (tentative)

Phenols & Chlorophenols EPA-8
                  (tentative)

Phenothiazine EPA-1 (tentative)

o-phenylphenol


Phorate EPA-1

Phoskil

Phosphorus Compounds EPA-1


Phygon

Picloram EPA-1  (tentative)

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 digestion and gravimetric
  procedure

Dichlone EPA-1

HPLC

UV (ether extraction)

UV (pyrophosphate extraction)

UV (water_-sp_luble_ formulations)

-------
                                31
Piperonyl Butoxide 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




Pomarsol




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)




Propargite EPA-2 (tentative)




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




Siraazine 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




Quaternary Ammonium 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




Ratlcate




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




Rumetan




Rutgers 6-12
Carbaryl EPA-1 & 2




Sodium Chlorate EPA-1




Norbormide EPA-1




dichlorprop




mecoprop




2,4,5-T




MH  EPA-1




Couraaphos 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 Hexariediol EPA-1  & 2

-------
S-276

666

Salicylanilide 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


Slmanex

slmazine

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-chlorophenbl

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

Siraazine 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

Streptomycin EPA-1

streptomycine  (France)

streptomycin hydrochloride

streptomycin nitrate

streptomycin sulfate

Strychnine 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 titration

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 spectrojscopy

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  sulfate  precipitation

CS2  extract^on^ (presence^ acetone-
   soluble pesticides)

-------
                                36
Sulfur Dioxide EPA-1

sulfurous acid anhydride

sulfurous oxide

Su Seguro 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 (tentative)

Terbutol EPA-2 (tentative)

Termil

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 & 2

IR

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-
  dicyanobenzene

cis-N-[(1,1,2,2-tetrachloroethyl)thio]-
  4-cyclohexene-l,2-dicarboximide

tetrachloroisophthalonitrile

0,0,0',O1-tetraethyl S,S'-raethylene
  bisphosphorodithioate

1,2,3,6-tetrahydro-3,6-dioxo-
  pyridazine

tetramethylthiuram disulfide

Thifor

Thimar

Thimet

Thimul

Thiodan

thiodemeton

thiodiphenylamine

Thionex

thiophal

thiophos

Thiram EPA-1

Thiram EPA-2

Thistrol

Thylate

Tillam

Tirabo (Brazil)

timet

tin,  organic  compounds

TMTD
Chlorothalonil EPA-1


Captafol EPA-1

Chlorothalonil EPA-1


Ethion EPA-1 & 2


MH  EPA-1

Thiram 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

Disulfoton 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

Thiram 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-(trichlorotnethylthio)phthalimide

2,4,5-trichlorophenoxy acetic acid

4-(2,4,5-trichlorophenoxy)butyric acid

2-(2,4,5-trichlorophenoxy)ethyl-2,2-
  dichloropropionate

2-(2,4,5-trichlorophenoxy)propionic
  acid

Tri-Endothal

or,a,
-------
                                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

Vergemaster

Vernam

Vernolate EPA-1

Vernolate 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-TCD-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 (sodium 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

-------
                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.

-------
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:
                             CH2CH3

                                           CH2 0
                                           C-CH2-C,
                            'CH2 CH3
Molecular formula:   C-.H.-C1NO.
                     14  20     2
Molecular weight:   269.8
Melting point:      39.5 to  41.5C
Physical state, color, and odor:  odorless, cream-colored crystalline
                                 solid  (at RT)
Solubility:  242 ppm in  water  at  25C; 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 ul
     4.  Usual laboratory glassware

Operating Conditions for TCD;
     Column temperature:     225C
     Injection temperature:  235C
     Detector temperature:   235C
     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 mg
     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.

     Calculation;
         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 B (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:


         v  (wt. I.S.)(% purity I.S.)(pk. ht. or area alachlor)(100)
             (wt. sample)(pk. ht. or area I.S.)(RF)
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 0CH3
Molecular formula:  C ,H  C1NO
Molecular weight:   269.8
Melting point:      39.5 to 41.5C
Physical state, color, and odor:  odorless, cream-colored crystalline
                                  solid (at RT)
Solubility:  242 ppm in water at 25C; 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
     A.  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. j 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.  Centrifuge or filtration equipment
     6.  Usual laboratory glassware

Operating Conditions for FID;
     Column temperature:     190C
     Injection temperature:  2AOC
     Detector temperature:   2AOC
     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 gran 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 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 triphenyl-
     me thane.
         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-
     me thane 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 = triphenylme thane
                 . 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 :

          B (wt. I.S.)(% purity I.S.)(pk. ht. or area alachlor) (100)
             (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:


                               -N
                               NH2
Molecular formula:  CCH,N
                     -> o
Molecular weight:   94.11



Melting point:      158C



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 water.  Mix thoroughly
     and pipette a 10 ml aliquot into a 100 ml volumetric flask.  Make
     to volume and again mix thoroughly,  (final cone 3 pg/ml)

     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 ug 4-amino-
     pyridine /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 302 run 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:
      ^ (abs. sample)(cone. std in jig/ml)(% purity std)
         (abs. std)(cone, sample in ug/ral)
Method submitted by  Stelios 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-amino-lH-l,2,4-triazole,
a registered herbicide having the chemical structure:-
                                 .r
                                 
                           N-
                  CNH2
Molecular formula:  C-H.N.
                     244
Molecular weight:
Melting point:
84.1
159C
Physical state, color, and odor:  white crystalline powder; odorless
                    when pure; bitter taste
Solubility:  soluble in water (28 g/100 ml at 25C);  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 chelatlng 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)5N0.2H20 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 /ig/ml)

-------
                            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 iig 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:

 = (abs. sample)(cone, std in ug/ml)(% purity std)
    (abs. std) (cone, sample in jig/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:
                     H2N	S  0 NH4
                               II
                               0
Molecular formula:  H,N00,S
                     o i 3
Molecular weight:   114.1
Melting point:      130C, decomposing at 160C;  the technical
                    product is at least 97% pure  and has a m.p.  of
                    131 to 132C.
Physical state, color, and odor:  Colorless, odorless, crystalline
                            I
                    solid (forms plates)
Solubility:  216 g/100 g water at 25C; soluble in glycerol, glycols,
             and formamlde; 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  	=-;	r^	
                       (grams sample)
     The milliequivalent weight of sodium nitrite for this determina-



     tion is 0.0230.





                       (69.01)    _  r
                          r
Reactions:



           tra  a. w=wn      _s. w-w  u  , 

                                  44    2
     NH4SO NH  + NaNO  	) NaNHSO  + N-t +
     2NaN02 + 2KI + AHC1	> I2 -- 2KC1 + 2NaCl + 2ND
     I- + 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.Cl.N.
                     95  34
Molecular weight:   275.5
Melting point:      159 to 160C
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

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                                                         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)

     De t ermina t ion;
         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"  (7.5 ji) .

-------
                                                       Anilazine EPA-1
                                                       (Tentative)
     Calculation:
         From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent anilazine as
     follows:

      _ (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.

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November 1975
Anilazine EPA-2
(Tentative)
                    Determination of Anilazine
                   by GasLiquid Chromatography
                     (TCD - Internal Standard)
     Anilazine is the common name for 2,4-dichloro-6-(o-chloroanilino)-
s-triazine, a registered fungicide having the chemical structure:
                        Cl

                        c
                   ^^
                 N

          Ct	C
                       N'
Molecular formula:  C_H_C1-N,
Molecular weight:   275.5
Melting point:      159 to 160C
                                              e
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)-1,3,5-triazin-
             2-amine

-------
                                 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.  6f 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:     215C
     Injection temperature:  230C
     Detector temperature:   230C
     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.

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                                                          Anilazlne 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 mg
     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 fpr each injection of the standard-
     internal standard solution as follows and calculate the average:

-------
                                                         Anilazine EPA-2
                                                         (Tentative)
        a (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 . (wt. HEOD)(% purity HEOD)(pk. ht. or area anilazine)(100)
         (wt. sample)(pk. ht. or area HEOD)(RF)
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 artenate 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 Na2H AsO, ; molecular weight 185.91; very soluble in
water, slightly soluble in alcohol; forms heptahydrate (7H?0)f
an odorless, crystalline solid that effloresces in warm air, loses
water and becomes anhydrous at 100C, forms pyroarsenate at 150C or
higher.  POISONOUS!

Sodium arsenite;
     Molecular formula approx. NaAsO,,; molecular weight 129.90; white
or grayish-white powder; somewhat hygroscopic; absorbs CO- from air;
freely soluble in water, slightly in alcohol; VERY POISONOUS!

Principle of the Method;
     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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                                                   Arsenic Compounds EPA-1
     The reaction:
                                           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:
NaAs02 + I2 + 3NaHC03  > Na2H As04 + 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







         S203 - *  Na2S46





Reagents ;




     1.  Iodine, 0.05N standard solution




     2.  Sodium thiosulfate, 0.05N standard solution




     3.  Concentrated hydrochloric acid, ACS




     A.  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

-------
                                 3                  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 NaAs02> 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:

     ,      .  _ (ml iodine) (N iodine) ( .03746) (100)
     /o cirsGnic a=           *          , *
                           (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.

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                            4                  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)
A arsenic = 	;	: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. 818C 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 HSO, 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.

-------
                            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  (MAA) will  give  an
idea of the general characteristics of  this group  of compounds:

                                                 .OH
Structural formula:
          "   /
CH3As<
    *j        \
                                               XOH
Molecular formula:  CH AsCL
Molecular weight:   140.0
Physical state, color, and odor:  odorless,  white,  crystalline solid
Melting point:  161C
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

     A.  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  O.lN standard  iodine solution to
     the first  permanent  blue  color.

     Calculation;
         Calculate the percent arsenic  as follows:

     v      .     (ml iodine)(N iodine)(0.03746)(100)
     /, arsenic  =  	-.	:r	
                           (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 i,t 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.

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                                 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)
     % arsenic * 	;	:7	
                            (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.

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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      ?       /CH3
    CH3-CH2 N - C^      C - N  CH<
                           ^NT                  XCH3
Molecular formula:   C-H.^Cl N5
Molecular weight:    215.7
Melting point:       173  to 175C
Physical state  and  color:  white crystalline solid
Solubility:  at 27C 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.)i 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

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                                                          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 mg atrazine/ml)

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                                 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 ^).
          Determine the absorbance of standard and sample using the
     peak at 1585 cm"1 (6.31 ju) and basepoint at 1675 cm"1 (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.

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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-
isopropylamino-l,3,5-triazine, a registered herbicide having the
chemical structure:

                               Cl
                                           V        /CH3
    CH3CH2N	O       C-	NOK'
                                                      CH3
Molecular formula:  C^H ,C1 N
Molecular weight:   215.7
Melting point:      173 to 175C
Physical state and color:  white  crystalline solid
Solubility:  at 27C 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

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                                 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 chroraatograph 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 jjl
     4.  Mechanical shaker
     5.  Centrifuge or filtration equipment
     6.  Usual laboratory glassware

Operating Conditions for FID;
     Column temperature:     190C
     Injection temperature:  240C
     Detector temperature:   240C
     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.

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                                                        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 rag
     atrazine arid 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 rag atrazine and 2 mg alachlor/ml)

     Determination;
         Inject 1-2 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 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.

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                                                          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:

     ., _ (wt. alachlor)(% purity alachlor)(pk. ht. or area atrazine)(100)
         (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.

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September 1975
                                       Azinphos-methyl EPA-1
                 Determination of Azinphos-methyl
                     by Infrared Spectroscopy

     Azinptibs-methyl is the common name for 0,O-d line thy 1  S-[4-oxo-l,2,3-
benzotriazin-3(4H)-y|methyl]phosphorodithioate,  a  registered  insecti-
cide having the chemical structure:
   CH30
\l
                PSCH2-N
   CH3-0'
Molecular formula:  C. H  N 0 PS
Molecular weight:   317.34
Melting point:      73 to 74C
Physical state and color:  white, crystalline solid
Solubility:  about 29 ppm in water at 25C; soluble in most organic
             solvents
Stability:   unstable at temperatures above 200C; 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

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                                 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
                                   *
     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 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-
     methyl 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)

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                                 3                 Azinphos-methyl EPA-1
     Determination;
         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 ji to 16.2 u).
         Determine the absorbance of standard and sample using the
     peak at 653.6 cm"  (15.3 /i) and basepoint 625 cm"  (16.0 yu).

     Calculation:
         From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent azinphos-methyl
     as follows:

      = (abs. sample)(cone, std in mg/ml)(% purity std)
         (abs. std)(cone, sample in mg/ml)

     (A concentration of 1 mg 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 u)
     basepoint:        784.9 cm"1 (12.74 u)
     cone:             12 mg/ml

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August 1975
                                     Benefin EPA-1
                                                                                B
                     Determination of Benefin
                     by Infrared Spectroscopy

     Benefin is the common name for N-butyl-N-ethyl-
-------
                                 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
         RFM 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 rag/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"  to 1240 cm"  (7.1 p to 8.1 ji) .
         Determine the absorbance of standard and sample using
     the peak at 1310 cm   (7.63  u) and a baseline from 1330 cm
     to 1260 cm"1 (7.52 p to 7.94 ^i) .

     Calculation:
         From the above absorbances and using the standard and
     sample solution concentrations, calculate the percent benefin
     as follows:

     a, _ (abs. sample) (cone. std  in mg/ml)(% purity std)
         (abs. std) (cone, sample in ing/ml)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services, 1 North 14th Street, Richmond,
Virginia 23219.

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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 N CH2-CH2-CH2-CH3
Molecular formula:   C,_H-,F_N_0.
                     13  lo 3 3 4
Molecular weight:    335.3
Melting point:       65  to 66.5C
            i
Physical state,  color,  and odor:  Yellow-orange crystalline solid
                    with no appreciable odor
Solubility:  70  ppm in  water at 25C; 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, Bahafin,
             Benalin, Binnell, Blulan, Bonalan, Carpidor, Quilan

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                                 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:            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.

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                                                         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 A 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.

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                                                        Benefin EPA-2
                                                        (Tentative)
         Determine the RF value for each injection of the standard-

     internal standard solution as follows and calculate the average:
        _    .  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 follows and calculate the

     average:


     7 _ (wt. diazinon)(% purity diazinon)(pk. ht. or area benefin)(100)
         (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.

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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
                      H_l_    _    _    _
                                H
                                     II
                           CNC0CH-
Molecular formula:  C ,HlgN,CL
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
Reagents;
     1.  Benomyl standard of known % purity
     2.  Chloroform, pesticide or spectro grade
     3.  Sodium sulfate, anhydrous, granular

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                                                        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)

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                                                        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  ) and basepoint at 1810 cm"  (5.52 i).
     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.

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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
                              CN C0CH3
Molecular formula:   C-.H,0N.0_
                     14  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:  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
Reagents;
     1.  Benomyl standard of known % purity
     2.  Dioxane, pesticide or spectro grade

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                                                      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 jig benomyl/ml)  Allox* to stand for three hours with
     occasional shaking.

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                                                         Benomyl EPA-2
                                                         (Tentative)
           Note:
              Benomyl absorbs strongly in the range of 260-310 run.
           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, however,
           that the peak at 294 nm was diminishing gradually until
           it practically disappeared.  Since this affects the peak
           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 sample 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:

       _ (abs. sample)(cone, std in ^ig/ml)(% purity std)
         (abs. std)(cone, sample in jig/ml)
Method submitted by Stelios Gerazounis, EPA Region II, New York, N. Y.

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,CH3
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:
                                                    /^
              OH                  S    ,0-CH<
               II    I                    II
              S-N-ChUrCt-U-S-p
               II          2     2
              0

Molecular formula:  C, ,H0.NO.PS-
                     14 24  4  3
Molecular weight:   397.5
Melting point:      34.4C (supercools readily)
Physical state and color:  colorless liquid or  white crystalline  solid
Solubility:  25 ppm in water at 20C; slightly  soluble in kerosene,
             moderately soluble in xylene, and  readily soluble in
             acetone and methanol
Stability:   relatively stable and non-corrosive; decomposes at
             elevated temperature over long periods of time  (at 80C
             in 50 hr   and at 200C in 18-40 hr)

Other names: Betasan - for turf use and Prefar  - for crop use (Stauffer);
             N-2-(0, 0-diisopropyl-phosphorothiolothionyl) ethyl benzene-
             sulfonamide;  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 ram 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 emulsifiable 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 ji).
         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 ;i) .

     Calculation;
         From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent bensulide as follows:
     y _ (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.

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February 1976
           BHC, gamma isomer EPA-1
                 Determination  of BHC, gamma isomer


             in Llndane  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
                      CI-CH
                      CI-CH
XH-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 112C.





 BHC, gamma isomer



 Molecular formula:  C,H,C1,
                     Do  D


 Molecular weight:   290.8



 Melting point:      112.9C



 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, Dolraix, FBHC, HCCH, Hexafor, Hexyclan,
             Soprocide

Reagents;
     1.  BHC, gamma isomer 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 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/ml)

-------
                                             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 spectro-
photometer at the optimum quantitative analytical settings, scan
both the standard and sample from 770 cm   to 650 cm   (13 11 to
                                                      -1
15.4 u).  Measure the absorbance of the peak at 687 cm   (14.55 u)
using a baseline from 720 era   to 673 cm"  (13.9 p to 14.85 ;i).

Calculation;
    Calculate the percent of BHC, gamma isomer as follows:

. _ (abs. sample)(cone. standard in mg/ml)(% purity standard)
    (abs. standard)(cone, sample in mg/ml)

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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:
                         =C CH=C CH-7
                                       CH-CH2CH3
                                       CH-
Molecular formula:  C1CH10N00,
                     1_> lo 2. o
Molecular weight:   322
Melting point:      68 to 69C
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 90A4 (Niagara);  FMC 9044;  Ambox,  dinoseb methacrylate

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                                 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 jppwd.er _pr_ _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)

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                            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 rag binapacryl/ml)
    For emulsifiable 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.  Allox^ 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 IR instrument
being used, scan both the standard and sample from 1540 cm   to
1220 cm"1 (6.5  j to 8.2  i) .
    Determine the absorbance of standard and  sample  using the peak
          -1
    346 cm   (7.
(7.10   to 7.85
          -1                                   -1           -1
at 1346 cm    (7.43  ) and baseline from 1408 cm   to 1273 cm
Calculation;
    From  the above absorbances and using the standard and sample
concentrations, calculate the percent binapacryl as follows:

y = ( ^b s .  samp 1 e ) ( cone . std in mg/ml)(% purity std)
    (abs.  std) (cone, sample in mg/ml)

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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:

                      Na2P407   10  H20
     Boric acid is a registered fungicide  and  insecticide having the
empirical formula:
                             H3B03
Borax:
Molecular formula:  Na2B,07 (anhydrous)
Molecular weight:
Melting point:
201.3 (anhydrous)
381.4 (decahydrate)
approx.  740C (anhydrous)
approx.  (enclosed  space)  62C  (decahydrate)
Physical state, color, and odor:  light gray odorless  solid  (anhydrous)
                                  white crystalline  odorless solid  (decahydrate)
Solubility:  in 100 ml water at 20C,  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 100C, 4 more at 160C, and becomes  anhydrous at 320C;
             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

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                                 2                    Boron Compounds EPA-1

Boric acid:
Molecular formula:  H BCL
Molecular weight:   61.84
Melting point:      approx. 160C
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 HBO_
             when heated at 100-105C; on long heating pyroboric acid
             H2B,07 is formed, and at higher temperatures the anhydride
             boric oxide B90_ 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)

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                                 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.

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                            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 any 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 NaOH)(N NaOH)(milliequivalent weight compound)(100)
                     (grams sample)(100/200)(100/200)
milliequivalent weights are:
    0.03482 for boric oxide B^
    0.06184 for boric acid H3B03
    0.05032 for sodium tetraborate, anhydrous Na?B,07
    0.09536 for sodium tetraborate, decahydrate Na       .

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                                 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.

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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:
                          H
NCHCH2CH3
              BrC,
                          11
                          0
Molecular formula:  CgH  BrN20_
Molecular weight:   261.1
Melting point:      158 to 159C
Physical state, color, and odor:  odorless, white crystalline solid
Solubility:  815 ppm in water at 25C; 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

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                                 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:  61 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 pi
     4.  Mechanical shaker
     5.  Centrifuge
     6.  Usual laboratory glassware

Operating Conditions for FID;
     Column temperature:     200C
     Injection temperature:  220C
     Detector temperature:   300C
     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 HSO,, 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 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.
         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.

-------
                                                      Bromacil EPA-1
                                                      (Tentative)
         Determine the RF value for each injection of the standard-

     internal standard solution as follows and calculate the average:


     PW o (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)(100)
         (wt. sample)(pk. ht. or area HEOD)(RF)
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.

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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
Molecular formula:  C,,H0Br_NCL
                     U o  J  z
Molecular weight:   449.96
Melting point:      227-228C
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-
             tivity

-------
                                                Brominated Salicvlanilides EPA-1
4',5-dibromosalicylanilide
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 Salicylanilides 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 tig 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.

Calculation:
    From the above absorbances and using the standard and sample
concentrations, calculate the percent polybrominated salicylanilide
as follows:

/ _ (abs. sample)(cone, std in ug/ml)(% purity std)
    (abs. std)(cone, sample in ug/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:
                       0          CH2CH	CH3
CH3CH2	SC	N
                                   CH2CH	CH3
 Molecular formula:   C  H-  ONS

 Molecular weight:    217.4

 Boiling point:      71C 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.  Chloroform, pesticide or spectro grade

-------
                                 2                     Butylate EPA-1
                                                       (Tentative)
Equipment:
     1.  Gas chromatograph with thermal conductivity detector (TCD)
     2.  Column:  51 x I/A" glass column packed with 20% SE-30 on
                                  
         Chromosorb W, AW, DMCS (or equivalent column)
     3.  Precision liquid syringe:  50 jil
     4.  Mechanical shaker
     5.  Centrifuge or filtration equipment
     6.  Usual laboratory glassware

     Operating Conditions for TCD:
         Column temperature:     180C
         Injection temperature:  2AOC
         Detector temperature:   270C
         Carrier gas:            Helium
         Flow rate:              100 ral/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:

     , _ (pk. ht. or area sample) (wt. std injected) (% purity of std)
     IQ 
         (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 name1for S-ethyl diisobutylthiocarbamate,
a registered herbicide having the chemical structure:
                                     CH2CH	CH3
  CHCH2  SC
                                     CH2CH	CH3
Molecular formula:   C   H   ONS
Molecular weight:    217.4
Boiling point:      71C 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.  Chloroform
     3.  Dichloromethane
     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 ^1



     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)
     Determination;

         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.

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September 1975                                          Butylate EPA-3
                                                       (Tentative)
                   Determination of Butylate
               by Gas-Liquid Chromatography  (FID)

     Butylate is  the common name for S-ethyl diisobutylthiocarbamate,
a registered herbicide having the chemical  structure:
                                    CH2	CH	CH3
  CH3CH2	SC	N
                                    CH2	CH	CH3
                                             CH3
Molecular formula:  C  -H.-ONS
Molecular weight:    217.4
Boiling point:       71C 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 Sampler
         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 jul 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          CH2 CH  CH3
CH3  CH2  S C - N<^
                                   CH2  CH - CH3
Molecular formula:  C  H  ONS
Molecular weight:   217. A
Boiling point:      71C 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.  S-Ethyl dipropylthiocarbamate (EPIC)  standard of known % purity
     3.  Carbon disulfide, pesticide or spectro grade
     A.  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 + 5% methanol.  (cone A mg EPTC/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:     130C
     Injection temperature:  225C
     Detector temperature:   250C
     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 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 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:

        = (wt. EPTC)(% purity EPTC)(pk. ht. or area butylate)	
          (wt. butylate)(% purity butylate)(pk. ht. or area EPIC)

         Determine the percent butylate for each injection of the
     sample-internal standard solution as follows and calculate the
     average:

     , _, (wt. EPTC)(% purity EPTC)(pk. ht. or area butylate) (100)
       = (wt. sample)(pk. ht. or area EPIC)(RF)
Method submitted by Division of Regulatory Services, Kentucky
Agricultural Experiment Station, University of Kentucky, Lexington,
Kentucky 40506.

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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,
 m
a registered herbicide having the chemical structure:


                                               CH3


                         0           CH2 CH - CH3

   CH3  CH2  S C - N<^

                                      CH2  CH - CH3
Molecular formula:   C11H23ONS

Molecular weight:    217.4

Boiling point:       71C 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  rag vernolate/ml)

-------
                                 2                       Butylate EPA-5
                                                         (Tentative)
Equipment:
     1.  Gas chromatograph with thermal conductivity detector (TCD)
     2.  Column:  5' x I/A" glass column packed with 5% PEG-1540
                  on 60/80 Chroraosorb W AW DMCS
     3.  Precision liquid syringe:  25 or 50 pi
     4.  Usual laboratory glassware

Operating Conditions for TCD;
     Column temperature:     150C
     Injection temperature:  200C
     Detector temperature:   200C
     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)(100)
    (wt. sample)(pk. ht. or area vernolate)(RF)

-------
                                                        Butylate 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. 321C, b.p. 767C, 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 H2SO,.  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 nm is used for macro amounts of
cadmium in formulations rather than the most sensitive absorption at
228.0 nm 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:
     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 maTrl.mmn  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 obeyed in the 100-500
     ppm range.

     Calculation;

         % Cadmium  = (PP^ Cd)(IQ"6)(100)
                     (.grams sampieMJ-/-LUu;
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-
thip)-4-cyclohexene-l,2-dicarboximide, a registered  fungicide having
the chemical structure:
                                    0
           HC           XHC          Cl   Cl
                                  \          I    I
                                    NSCCH
           HC\        /CH-C^         ci   ci
                                  X0
Molecular formula:   C,0H-C1^NO-S
Molecular weight:    349.1
Melting point:       160  to 161C
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

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                                                         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:
     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 mg/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 flovable 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)

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                                                          Captaf ol EPA-1
                                                          (Tentative)
     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 2000 cm   to
     1540 cm"1 (5.0 ji-6.5 ;i).
         Determine the absorbance of standard and sample using the
     peak at 17.27 cm   (5.79 ji) and baseline from 18.18 cm   to
     1639 cm'1 (5.5 p. to 6.1 ;i).
     Calculation;
         From the above absorbances and using the standard and sample
     concentrations, calculate the percent captafol as follows:

      a (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.

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September 1975
                                      Cap tan EPA-1
                   Determination of Captan by
                 the  Hydrolyzable Chlorine Method

     Captan is the common name for N-trichloromethylthio-A-cyclo-
hexene-l,2-dicarboximide, a registered fungicide having the chemical
structure:
               V2
              -C
      HC
      HC
              0
.CH-
      Cl
      I
-sc
      I
      Cl
                                               a
               HZ
Molecular formula:   C0H.C1,NO,S
                     y o  j  i
Molecular weight:   300.6
Melting point:      178C  (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.  160-170C
Solubility:  less than 0.5 ppm in water at RT; insoluble in petroleum
             oils;  at 25C 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.), Herpan, Vondcaptan

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                                 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.)

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                            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 30%
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. Cl_) from the
                                                   o
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.

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                                                  Captan EPA-1
  captan  -  (meq* C1T " oeq' CV <-1002> <100>
             (weight sample)(0.4)
where:  meq. Cl_    N AgNO, X ml used for titration of

                     hydrolyzed aliquot

        meq. Cln  "  N AgNO. X ml used for titration of
               D           J
                     non-hydrolyzed aliquot


        meq. wt. captan   =   (3)(100Q)  "  -1002


        aliquoting factor     yr     0.4

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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
                                        1 S	C	Cl
                                                   Cl
                                  0
Molecular formula:  C H Cl
Molecular weight:   300.6
Melting point:      178C  (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. 160-170C
Solubility:  less than 0.5 ppm in water at RT;  insoluble in petroleum
             oils; at 25C 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

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                                 2                       Captan EPA-2
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 0.1 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.

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                                 3                         Cap tan EPA-2
     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 1885 cm   to*
     1665 cm"  (5.3 ji to 6.0yu).
         Determine the absorbance of standard and sample using the
     peak at 1735 cm"  (5.76 ^i) 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) (% purity std)
         (abs. std)(cone, sample in mg/ml)

     (A concentration of 1 mg captan/ml 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.

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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:
                                      0
                                       II
H
                                0C	N  CH3
Molecular formula:  Ci2HnN02
Molecular weight:   201.2
Melting point:      142C
Physical state and color:  white, crystalline solid
Solubility:  40 ppm in water at 30C; 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.

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                                 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 jig carbaryl/ml)

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                            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:

 = (abs. sample)(cone. std in ug/ml)(% purity std)
    (abs. std) (cone, sample in

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October 1975
Carbaryl EPA-2
(Tentative)
                   Determination of Carbaryl by
               High Pressure Liquid Chromatography

     Carbaryl is the accepted common name for 1-naphthyl methylcar-
bamate, a registered insecticide having the chemical structure:
                                      0     H
                                      II
                                0	C	N  CH3
Molecular formula:  C12H11N2
Molecular weight:   201.2
Melting point:      142C
Physical state and color:  white, crystalline solid
Solubility:  40 ppm in water at 30C; soluble in most polar organic
             solvents such as acetone, dlmethylformamide
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

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                                 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 Permaphase, 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:  50-55C
     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.

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                                                        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:

      _ (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.

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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:  ci2H15N03
Molecular weight:   221.3
Melting point:      150-152C
Physical state, color, and odor:  Odorless,  white,  crystalline solid
Solubility:  solubility at 25C is 700 ppm in water,  15% in acetone,
             14% in acetonitrile, 4% in benzene,  9% in cyclohexanone,
             27% in dimethyIfonnamide
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

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                                 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)

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                                                      Carbofuran EPA-1
     Determination:
         With chloroform in the reference cell, and using the optimum
     quantitative analytical settings, scan both the standard and
         ile from 1000 cm   to 800 cm   (10 p to 12.5 ji).
         Determine the absorbance of standard and sample using the
         : at 875 cm"1
     (11.11 i to 11.83
sample from 1000 cm"  to 800 cm   (10 p to 12.5
              the absorbance of standard and sa
peak at 875 cm"1 (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.

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 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-tnethyl-l,4-oxathiin-
3-carboxanilide, a registered fungicide having the chemical structure:
Molecular formula:  C  H  NO-S
                     \-* J. J  
Molecular weight:   235
Melting point:      91.5 to 92.5C; a dimorphic form has a m.p. of 98 to 100C
Physical state, color, and odor:  odorless, white, crystalline solid
                    (The technical product is at least 97% pure.)
Solubility:  170 ppm in water at 25C; 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 spectre grade
      3.  Sodium sulfate, anhydrous, granular

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                                                       Carboxiri EPA-1
                                                       (Tentative)
Equipment:
      1.  Infrared spectrophotometer, double beam ratio recording,
          with matched 0.2 ram 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)

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                                                       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 fi 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 ;i)         1630 cm"1 (6.13^)
              1585 cm"1 (6.30 p)         1630 cm"1 (6.13^i)
              1290 cm"1 (7.75 ji)         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:

      = (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.

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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:
                                                  0CH3
Molecular formula:  C H  BrCIN 0
Molecular weight:   293.6
Melting point:      97C (The technical grade has a purity of 95%
                          and melts at 90-95C)
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)

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                                 2                     Chlorbromuron EPA-1
                                                       (Tentative)
Reagents;
     1.  Chlorbromuron standard of known % purity
     2.  Acetone, pesticide or spectre grade

Equipment:
     1.  Gas chromatograph with flame ionization detector (FID)
     2.  Column:  2' x I/A" glass column packed with 2% SE-52 on
         70/80 Anakrom ABS (or equivalent column)
     3.  Precision liquid syringe:  10 jil
     A.  Mechanical shaker
     5.  Centrifuge or filtration equipment
     6.  Usual laboratory glassware

     Operating Conditions for FID:
         Column  temperature:     180C
         Injection temperature:  225C
         Detector temperature:   225C
         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)

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                                                     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 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 Chlorbromuron as follows:

      _ (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.

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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-D (ISO, BSI, WSSA),    2,4-dichlorophenoxyacetic acid



                               Cl
                                     0-CH2-COOH
Molecular formula:  CQH,C1_0_
                     o o  / J


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 141C (pure), 135 to 138C (technical)
Solubility:
Stability:
Other names:
about 600 ppm in water at 25C;  soluble in aqueous
alkali and in alcohols, ether,  acetone; insoluble in
petroleum oils
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, Ded-Weed, Dormone,
Esteron, Estone, Fernesta, Feminine, 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
                                     0CHCOOH
Molecular formula:  C0H0C1_0_
                     y o  JL 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 118.1C (pure), 114 to 117C (technical)
Solubility:  about 350 ppm in water at 20C; soluble in most organic
             solvents

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                                3            Chlorophenoxy Herbicides EPA-1


Stability:     acid  is stable to heat and resistant to reduction, hydrolysis,
              and atmospheric oxidation
Other names:   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
                                0CH2-CH2-CH2-COOH
Molecular formula:   CioHioC123
Molecular weight:    249.1
Physical state, color,  and odor:  white crystalline solid,  odorless when
                                 pure
Melting point:  117 to  121C 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

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                                           Chlorophenoxy Herbicides EPA-1
2,4,5-T (ISO, BSI, WSSA),     2,4,5-trichlorophenoxy acetic acid

                               .Cl
                                      0CH2-COOH
Molecular formula:  C0H Cl,0_
                     O J  J J
Molecular weight:   255.5
Physical state and color:  white crystals
Melting point:  156.6C (pure),  150-151C  (technical)
Solubility:     about 278 ppm  in water  at  25C; 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
Stability:
Other names:
stable and non-corrosive
Weedone 2,4,5-T (Amchem),  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
                                         CHCOOH
                                          CH3

-------
                                         Chlorophenoxy Herbicides EPA-1
Molecular formula:   C  H  Cl  0
Molecular weight:    269.5
Physical state,  color, and  odor:  white powder, low odor
Melting point:   179  to 181C
Solubility:


Stability:
Other names:
2.4.5-TB (ISO),
about 140 ppm in water at  25C;  soluble  in acetone
and methanol
non-corrosive to spray equipment
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

   4-(2,4,5-trichlorophenoxy) butyric acid
          Cl
                               0- CH2- CH2- CH2.-COOH
Molecular formula:   C  H Cl 0,
Molecular weight:   283.5
Physical state and  color:  white  crystals
Melting point:  114 to 115C
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
                                     0-CH2COOH
Molecular formula:  Cg
Molecular weight:   200.6
Physical state and color:  white crystalline  solid  (pure), light
                           brown solid  (technical)
Melting point:  118 to 119C (pure),  100  to 115C  (technical)
Solubility:  about 825 pptn 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-A-chlorophenoxy) propionic acid
                                      0 CHCOOH
                                           CH3

-------
                                         Chlorophenoxy Herbicides EPA-1
Molecular formula:   C.-H  CIO.
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 95C (technical 90C or above)
                about 620 ppm in  water  at  20C; 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
Solubility:

Stability:

Other names:
MCPB (WSSA),    4-(2-methyl-4-chlorophenoxy)  butyric  acid
                                0-CH2-CH2-CH2-COOH
Molecular formula:  C .H.-C10-
Molecular weight:   228.5
Physical state and color:  white solid
Melting point:  100 to 101C (pure), 99 to 100C (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-CC-CH3
                                                     Cl
Molecular formula:   C11H_C1,0.
Molecular weight:    366.5
Physical state and  color:  white solid (pure), dark brown solid  (technical)
Melting point:  49  to 50C; bp 161 to 164C 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
                0-CH2-CH20
Cl
-/Vo-
            tris  form
           bis form
Molecular formula:  C0/H  Cl 0 P (tris), C.,H11.C1/0,P (bis)
                     ZH ^l  o D           ID ID  A D
Molecular weight:   6A9.4 (tris), 460 (bis)
Physical state,  color, and odor:  dark amber viscous liquid  with a
                                 phenolic odor
Boiling point:   above 200C 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

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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 jig 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

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                                 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 40C.  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 jig 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
     or 289 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.IN 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.

      = (abs. sample) (cone, standard in jjg/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.

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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

-------
                                      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:  55C
     3.  Pressure:     700-1000 psi (DuPont - constant pressure)
     4.  Flow rate:    0.5 to 1.5 ml/tnin (Perkin-Elmer - constant flow)
     5.  Chart speed:  5 minutes/inch or equivalent
     6.  Attenuation:  adjust for 60-80% pen response for 5 pi 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:

     '/ = (pk. ht. or area sample) (wt. std injected) (% purity of std)
         (pk. ht. or area standard)(wt. sample injected)

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                                 4   Chlorophenoxy Herbicides EPA-3
                                                        (Tentative)

Notes:

     1.  If a peak for a declared acid herbicide does not appear

         using the 10% methanol-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,

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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 ul
     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:     175C
                                  
     Injection temperature:  225C
     Detector temperature:   225C
     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.A 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 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

        m (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)(100)
         (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.

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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 derlvatization)

     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-
phenpxy 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-tr-imethylsilyl-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 chrotnatograph with flame ionization detector (FID)
     2.  Column:  6' x 4 mm 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 245C at 4/minute,
                             hold 4 minutes at final temp, of 245C
     Injection temperature:  250C
     Detector temperature:   250C
     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 mg 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)

Determination:
    Injections are made with the syringe filled as follows:  0.5 ^il
acetone, 0.5 ;il air, 1.0 jil MSTFA, 0.5 pi air, 2 pi of either
standard or sample.  Inject 2 ul 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.

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                                               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:

     % tt (wt. IS)(% purity IS)(pk. ht. or area CPH)(100)
         (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.

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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:
Molecular formula:  C-C1.N
Molecular weight:   266
Melting point:      250 to 251C
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 ppra); 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 Chem. Co.); Bravo; Termil;
             2,A,5,6-tetrachloro-l,3-dicyanobenzene; 2,4,5,6-tetra-
             chloro-3-cyanobenzonitrile

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                                 2                    Chlorothalonil EPA-1

Reagents;
     1.  Chlorothalonil standard of known % purity
             \
     2.  Methylene chloride, pesticide or spectro grade
     3.  Sodium sulfate, anhydrous, granular

Equipment:
     1.  Infrared spectrophotometer, double beam ratio recording
         with matched 0.5 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 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

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                                 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 flovable 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
     1R instrument being used, scan both the standard and sample from
     1050 cm"1 to 900 cm"1 (9.5 ji to 11.1 ;j).
         Determine the' absorbance of standard and sample using the
     peak near 980 cm   (10.2 u) and a baseline from 1000 cm   to
     940 cm"1 (10 ji to 10.64 p) .

     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.C. 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.

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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,	c.          c	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;
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 t
     1.  Potentiometric titrimeter with a silver electrode and a
         silver-silver chloride electrode
     2,  Steam bath
     3.  Usual laboratory glassware

Prpcedure;
     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 -  (1 AgTO3)(N AgN03)(.03545)(100)
                                    (grams sample)

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                            3           Chloro-Triazlne 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 AgN03)(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

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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:  c^E

Molecular weight:   290.7

Melting point:      149 to 150C

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,

             Chloroxlfenidim


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 ji).
         Determine the absorbance of standard and sample using the
     peak at 1351 cm'1 (7.40 ji) and basepoint at 1316 cm"   (7.60 ji).

-------
                                                      Chloroxuron EPA-1
                                                      (Tentative)
     Calculation:
         From the above absorbances and using the standard and

     sample solution concentrations, calculate the percent
     chloroxuron 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 Product Analysis Laboratory,

San Francisco, California.

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October 1975
Chloroxuron
(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 150C

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 yul
     4.  Usual laboratory glassware

Operating Conditions for TCD;
     Column temperature:     170C
     Injection temperature:  200C
     Detector temperature:   200C
     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)

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                                                       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 rag chloroxuron and 10 mg dieldrin/ml)

     Determination:
         Inject 10-20 ul 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)(100)
         (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.

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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 122-124C;  technical 119-120C
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,P207.10 H20 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 jig/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                      Goumafuryl EPA-1
solution, and shake on a mechanical shaker for one hour.  Transfer
30-40 ml to a glass-stoppered centrifuge tube 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 ug 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 ug/ml)(% purity std)
    (abs. std) (cone, sample in jig/ml)

or using dilution factors, as follows:

  m (abs. sample)(wt. std)(purity std)(1/100)(5/50)(5/50)(100)
  = (abs. std)(wt. sample)(1/50)C5/50)CO/10)

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November 1975
               Coumafuryl EPA-2
           Determination of  Coumafuryl  in  Concentrates
                   by Ultraviolet  Spectroscopy

     Coumafuryl is a common  name for  3-(alpha-acetonylfurfuryl)-A-
hydroxycoumarin, a registered  rodenticide  having  the chemical structure:

                          OH
                                         c=o
Molecular formula:  C-7H ,0
CH3
Molecular weight:   298.3
Melting point:      pure 122-124C;  technical 119-120C
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
                           r
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     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.  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 vrith 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 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 injng/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)(2/10)

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  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:
CH3CH20
CH3CH20
  Molecular formula:  C ,H  CIO PS
  Molecular weight:   362.8
  Melting point:
pure - 95C;  technical  -  91  to  92C
  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 1R
instrument being used, scan both the standard and sample from
1430 cm"1 to 1110 cm'1 (7 jj-9 -;i).
     Determine the absorbance of standard and sample using the
peak at 1277 cm   (7.83 u) and baseline from 1307 cm"  to
1227 cm"1 (7.65 p to 8.15 p).

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)
    (abs. std)(cone, sample in mg/ml)

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  September 1975
                                      Coumaphos EPA-2
                                      (Tentative)
                      Determination of Coumaphos
                by High Pressure  Liquid Chromatography

       Coumaphos is the common name for 0,0-diethyl  0-(3-chloro-4-
  methyl-2-oxo-2H-l-ben2opyran-7-yl)  phosphorothioate, a registered
  insecticide having the chemical structure:
CH3CH2	0
    -~j
\
                       P0
CH3CH20'
  Molecular formula:  C ,H -C1CLPS
  Molecular weight:   362.8
  Melting point:
   pure - 95C; technical - 91 to  92C
  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 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:        40% methanol + 60% water
         Column temperature:  50-55C
         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 coumaphos 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 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 coumaphos 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.

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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:
CH3CH2 0
CH3CH2  0'
                          0
Molecular formula:  C ,H  C105PS
Molecular weight:   362.8
Melting point:      pure - 95C; technical  -  91  to 92C
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:     210-220C
     Injection  temperature:  250C
     Detector temperature:   250C
     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 coumaphos 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 pi 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.

-------
                                                        Couraaphos  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)(100)
         (wt. sample)(pk. ht. or area tetradifon)(RF)
Method submitted by Division of Regulatory Services, Kentucky
Agricultural Experiment Station, University of Kentucky, Lexington,
Kentucky 40506.

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November 1975
   Crufomate EPA-1
   (Tentative)
                    Determination of Crufomate
                     by Infrared Spectroscopy

     Crufomate is the accepted common name for  A-tert-butyl-2-chloro-
phenyl methyl methylphosphoramidate, a registered insecticide  and
helminthicide having the chemical structure:
                 9*3
0CH3
         CH3-C(/
Molecular formula:  C  H  C1NO P
Molecular weight:   292.1
Melting point:  60C; technical product b.p.  117  to 118C  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 60C 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 crufomate/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 emulsifiable 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 60C before making to volume.)   Add a small
     amount of granular anhydrous sodium sulfate to  insure dryness and
     clarify the solution.  (cone 10 mg crufornate/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 crufornate/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"1 to 900 cm"1 (7.0 u to 11.0 ji) .
         Determine the absorbance of standard and sample using the peak
     at 1042 cm"  (9.60 u) and a baseline from 1333  cm"  to, 1000 cm"
     (7.50 ^ to 10 p).

     Calculation:
         From the above absorbances, calculate the percent crufomate as
     follows:
     ~ = (abs. sample)(cone. std in ing/ml)(% purity std)
         (abs. std) (cone, sample in nig/ml)

This method is adapted from Dow Chemical Company method no. 72733,
September 20, 1965.

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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:
                                               0   0-CH3

                                                    N_CH3
                                                    H
Molecular formula:  C12H
Molecular weight:   292.1
Melting point:  60C; technical product b.p. 117 to 118C 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                     Crufpmate EPA-2
                                                       (Tentative)
Reagents:
     1.  Crufornate 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 (TGD)
     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:     170C
     Injection temperature:  200C
     Detector temperature:   200C
     Filament current:       225C
     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
     crufomate and 20 mg HEOD/ml)
                                                                  t
     Preparation of Sample:
         Weigh a portion of sample equivalent to 0.1 gram crufomate
     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 crufomate.  For
     coarse or granular materials, shake mechanically for 30 minutes
     or shake by hand intermittently for one hour.  (final cone 10 rag
     crufomate and 20 mg HEOD/ml)

     Determination:
         Inject 5 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 crufomate, 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.

-------
                                                       Crufomate 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. crufomate)(% purity crufornate)(pk. ht. or area HEOD)
         Determine the percent crufomate for each injection of the
     sample-internal standard solution as follows and calculate the
     average:

        (wt. HEOD)(% purity HEOD)(pk. ht. or area crufomate)(100)
      e (wt. sample)(pk. ht. or area HEOD)(RF)
Method submitted by David Persch and George Radan, EPA Region II,
New York, N. Y.

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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
                   H
   CH3CH2N
\
                 H    CH3
                 I      I
           C N C  CBN
                               N'
Molecular formula:  C H  C1N,
Molecular weight:   240.7
Melting point:      166.5 to 167C
Physical state and color:  white crystalline solid
Solubility:  at 25C 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   to 930 cm"  (9.1 p to 10.8 ^i).
         Determine the absorbance of standard and sample using the
     peak at 1060 cm"  (9.43 ji) and basepoint 955 cm"  (10.47 p).
     Calculation;
         From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent cyanazine as
     follows:

      a (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.

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 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:
                                          CH2Chb
                                                   \
                          0                          ,CH2
CH3	CH2	SCN<^       CH2CH2

                                      CH2
 Molecular formula:   C.  H_ NOS

 Molecular weight:    215.A

 Boiling point:      1A5C 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 jul
     4.  Mechanical shaker
     5.  Centrifuge or filtration apparatus
     6.  Usual laboratory glassware

     Operating Conditions for TCD;
         Column temperature:     210C
         Injection temperature:  240C
         Detector temperature:   270C
         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)

-------
                                 3                      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 jal 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:

     , = (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.

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 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           /CH

CH3	CH2	S  C  N<(       CH2-CH2

                                          :CH3
 Molecular formula:   C  H  NOS

 Molecular weight:    215.A

 Boiling point:      145C 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

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                                 2                    Gycloate EPA-2
                                                      (Tentative)
Equipment;
     1.  Gas chromatograph with flame ionization detector (FID)
     2.  Column:  6' x 1/4" glass column packed with 3% OV-1 on
         80/100 Gas Chrom Q (or equivalent column)
     3.  Precision liquid syringe:  10 jil
     4.  Mechanical shaker
     5.  Centrifuge or filtration apparatus
     6.  Usual laboratory glassware

     Operating Conditions for FID;
         Column temperature:     175C
         Injection temperature:  225C
         Detector temperature:   220C
         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/ml)

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                                                      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 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 cycloate as follows:

      = (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.

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October 1975                                             Cycloate EPA-3
                  Determination of  Cycloate by
                   Gas-Liquid Chromatography
                   (FID - Internal  Standard)

     Cycloate  is the common name for S-ethyl cyclohexylethylthio-
carbaraate,. a registered herbicide having the chemical structure:
                                          CH2-CH2
                                                    VCH2
                         if       /V.    -./
CH3CH2
                         0           ,CH
                    	CN<       CH2-CH2
                                          CH3
Molecular formula:  C  H  NOS
Molecular weight:   215.4
Boiling point:      145C 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.  Pebulate standard of known % purity
     3.  Carbon  disulfide, pesticide or spectro grade
     4.  Chloroform, pesticide or spectro grade

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                                 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 11!
     4.  Mechanical shaker
     5.  Centrifuge or'filtration equipment
     6.  Usual laboratory glassware

Operating Conditions for FID;
     Column temperature:     140C
     Injection temperature:  225C
     Detector temperature:   250C
     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.

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                                                            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 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 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.

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                                                            Cycloatc 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:

      _ (wt. pebulate)(% purity pebulate)(pk. ht. or area cycloate)(100)
         (wt. sample)(pk. ht. or area pebulate)(RF)
Method submitted by Division of Regulatory Services, Kentucky

Agricultural Experiment Station, University of Kentucky, Lexington,

Kentucky 40506.

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December 1975                                           Dalapon EPA-1

                     Determination of Dalapon           .                        r\
                     by Infrared Spectroscopy

     Dalapon is the accepted common name for 2,2-dichloropropionic
acid, a registered herbicide having the chemical  structure:

                                 ci    v
                                  I
                        CH3C-COOH
                                 CI
Molecular formula:  C.H.C1.0-
Molecular weight:   143
Boiling point:      185 to 190C
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 (spectroscoplc grade preferred)
     3.  Sulfuric acid, 1+3
     4.  Anhydrous sodium sulfate, ACS granular

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                                                         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 A 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 A 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 11.0 u) using
     carbon disulfide in the reference cell.  For qualitative comparison,
     run a full scan.

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                                                            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 ji to 9.85 ;i)>
     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.

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January 1976
                                                           DDT   EPA-1
        Determination of DDT in Etnulsifiable 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:
Molecular weight :
Melting point:
                    354 . 5
                    108. 5C
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

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                                 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,
              1,1,1-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 60C.  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"  (8.5ji to 10.5 ji) .
    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 ji
to 10.3 ji) .

Calculation;
    From the above absorbances and using the standard- and sample
concentrations, calculate the percent technical DDT as follows:

     _ (abs. sample)(cone. std in mg/ml)(% purity std)
        (abs. std)(cone, sample in mg/ml)

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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:
                                             CH2-CH3
Molecular formula:  C  H  NO
Molecular weight:   191.3
Boiling point:      111C 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.

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                                 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 Nad or KBr cells
     2.  Rotary evaporator
     3.  Water bath at 50C
     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  insure dryness.  (cone 40 mg/ml)

     Preparation of Sample;
         Weigh a portion of sample  (alcohol solution and aerosol non-
     volatile) equivalent to 6.4  gram deet into a 125 ml Erlenmeyer
     flask and evaporate the alcohol under vacuum on a rotary evaporator
     at  50C.  (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 arid 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)

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                                                          Deet EPA-1
                                                          (Tentative)
     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
     770 cm"1 to 665 cm"1 (13 ja to 15 ^) .
         Determine the absorbance of standard and sample using the
     peak at 706.7 cm   (14.15 ji) and basepoiht 692.5  cm"  (14.44yu).

     Calculation:
         From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent deet as follows:

         , _ (abs. sample) (cone, std in mg/ml) (% purity std)
             (abs. std)(cone, sample in rag/ml)
         The above method is based on the old USDA, PRO 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   (11.6^1) 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.

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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:
Molecular formula:  C -H _NO
Molecular weight:   191.3
Boiling point:      lll'C at 1 mm Hg
Physical state and color:  colorless to amber liquid, nearly odorless;
                    the technical product contains 85-95% ra 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.

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                                 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 grains 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 jil
     4.  Usual laboratory glassware

Operating Conditions for TCD:
     Column temperature:     190C
     Injection temperature:  215C
     Detector temperature:   215C
     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.

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                                                           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)

     Determination;
         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:

     RF = (wt. heptachlor)(% purity heptachlor)(pk. ht. or area deet)
          (wt. deet)(% purity deet)(pk. ht. or area heptachlor)

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                                                           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)(pk. ht. or area deet)(1001
         (wt. sample)(pk.  ht.  or area heptachlor)(RF)
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.

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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:      111C 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

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                                 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:  4' 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:     150C
     Injection temperature:  200C
     Detector temperature:   200C
     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.

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                                                           Deet EPA-3
                                                           (Tentative)
Procedure:
     Preparation 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 jal 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:

     Rp = (wt. vernolate)(% purity vernolate)(pk. ht. or area deet)
          (wt. deet)(% purity deet)(pk. ht. or area vernolate)

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                                                          Deet EPA-3
                                                          (Tentative)
         Determine the percent deet for each injection of the sample-

     internal standard solution as follows and calculate the average:
         (wt. vernolate)(% purity vernolate) (pX-L h_t .  or area deet) (100)
         (wt. sample) (pk. ht .  or area vernolate) (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. 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.

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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
           CH2:0'
                           P	0
                         N
Molecular formula:  C  H  N 0 PS
                     \~* a. . j
Molecular weight:
Boiling point:
304.3
83 to 84C 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 120C; 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

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                                 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:  41 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:     170C
     Injection temperature:  200C
     Detector temperature:   200C
     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 emulsiftable 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 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
     diazinon as follows:

      a (pk. ht. or area sample)(wt. std injected)(% purity of std)
         (pk.,ht. or area standard)(wt. sample injected)

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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 o,0-diethyl 0-(2-isopropyl-6-methyl-4-
pyrimidinyl) phosphorothioate; a registered insecticide,  acaricide,
and nematocide having the chemical structure:
  CH3	CH20
 CH3	CH20
Molecular formula:  Ci2H21N23PS
Molecular weight:   304.3
Boiling point:      83 to 84C 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 120C; 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

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                                 2                    Diazinon EPA-2
                                                      (Tentative)
Other names:  Spectracide, Diazinon, G-24480 (Ciba-Geigy);  Basudin,
              Diazajet, Diazide, Diazol, Dazzel, Gardentox, Neocldol,
              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 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:        20% methanol + 80% water
         Column temperature:  50-55C
         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.

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                                                      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 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
     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.

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No vember.,1975 ,
                                    Diazlnon EPA-3
                    Determination of  Dlazinon
                     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
  CH'3 -CH2 0
 CH3 - CH2  0'
  xyn
304.3
Molecular formula:  C
Molecular weight:
Boiling point:      83 to 84C 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 ppro) in water at RT;  miscible  with  aliphatic and
             aromatic solvents, alcohols, and ketones; soluble  in
             petroleum oils
Stability:   decomposes about 120C; 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

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                                 2                     Diazinon EPA-3
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.  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 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 rag diazinon/ml)

     Determination:
         With acetone in the reference cell, and using the optimum
     quantitative analytical settings, scan both the standard and
     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 ;i) 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:

     ~ = (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.

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November 1975
Diazinon 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
 CH3	CH2	0
         CH3
Molecular formula:  Ci2H2iN23PS
Molecular weight:   304.3
Boiling point:      83 to 84C 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 120C; 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

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                                 2                     Diazinpn EPA-4
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.  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 rag 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 Ghrom 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:     175C
     Injection temperature:  260C
     Detector temperature:   255C
     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)

     Determination:
         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 elutlon 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:


        = (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:


     % m (wt. HHDN)(% purity HHDN)(pk. ht. or area diazinon)(100)
         (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.

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November 1975
              Dibromochloropropane EPA-1
              Determination of Dibromochloropropane
                     by Infrared Spectroscopy

     Dibromochloropropane is the trivial name for  l,2-dibromo-3-chloro-
propane, a registered soil fumigant and nematocide having  the  chemical
structure:
                H-
                        H
H
H
                         Br     Br     Cl
                 H
Molecular formula:  C_H Br.Cl
Molecular weight:   236.3
Boiling point:      196C
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.) Neraagon (Shell Development Co.)>
             DBCP, Nemafume, BBC 12, OS 1897

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                                 2              Dibromochlorbpropane EPA-1
Reagents:
     1.  Dibrpmochloropropane 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
                        -  *        '          
     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.
                                                           0
               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)

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                            3                Dibromochloropropane EPA-1
Preparation of Sample:
    For emulsiftable 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/ral)
    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 dryness.
(final cone 50 mg dibroraochloropropane/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 500 cm   (12.5 p to 20 u).
    Determine the absprbance of standard and sample using the peak
at 572 cm"  (17.48 ji) and baseline from 610 cm   to 520 cm   (16.4 p
to 19.2 jj) .

Calculation:
    From the above absorbances and using the standard and sample
concentrations, calculate the percent dibromochloropropane as
follows:

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                                                   Dibromochloropropane EPA-1
         (abs. sample)(cone, std in mg/ml)(% purity std)
         (abs. std)(cone, sample in mg/ml)
         (A concentration of 1 mg dibromochloropropane/ml carbon
          disulfide gives an absorbance of approx. 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.

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November 1975
                        Dibromochloropropane EPA-2
                        (Tentative)
              Determination of Dibroraochloropropane
                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
C
         H
H
C
                         Br     Br     Cl
H
Molecular formula:  C_H5Br2Cl
Molecular weight:   236.3
Boiling point:      196C
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

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                                 2                 Dibromochloropropane EPA-2
                                                   (Tentative)
Reagents:
     1.  Dibromochloropropane standard of known % purity
     2.  Chloroform, pesticide or spectro grade

Equipment:
     1.  Gas chroraatograph 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 ^il
     4.  Mechanical shaker
     5.  Centrifuge or filtration equipment
     6.  Usual laboratory glassware

Operating Conditions for TCD:
     Column temperature:     140C
     Injection temperature:  175C
     Detector temperature:   175C
     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:
         Weigh 0.20 gram dibromochloropropane standard into a 10 ml
     volumetric flask and make  to volume with chloroform.  (final cone
     20 mg/ml)

-------
                                                 Dlbromochloropropane KPA-2
                                                 (Tentative)
     Preparation of Sample:
         For technical material and liquid formulations,  weigh a portion
     of sample equivalent to 0.20 gram dibromochloropropane 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 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
     dibromochloropropane 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.

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January 1976                                     Dibutyl Succinate EPA-1


               Determination of Dibutyl Succinate
                by Saponification and Titration


     Dibutyl  succihate is a  registered insect  repellent with the  following

chemical structure:

              0
              II
     CH2	CoCH2CH2  CH2	CH3
              0
              II
     CH2 C 0CH2	CH2  CH2CH3
Molecular formula:  CioHooO,

Molecular weight:   230.3


Melting/boiling point:  m.p.-29C; b.p. 108C at  4 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)7 
COOH

-------
                                 2                   Dibutyl Succinate EPA-1

     Molecular formula:   C10H_.0.       Molecular weight:  282.45
                          lo 34 i
     Colorless liquid; solidifies at 4C 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 hydrolysis
         A =  (ml HC1 for Blank-mi HC1 for Sample)(N HC1)

     B = milliequivalents of free acid before hydrolysis
         B =  (ml NaOH)(N NaOH)

     ,  ^    ,              (A - B)(0.11515)(100)
     % Dibutyl succinate = *-.	"	r^	L
            3                 (grams sample)
          (milliequivalent weight of dibutyl succinate = 0.11515)

     % Oleic  acid =  (BH0.28215U100)
                     (grams sample)
          (milliequivalent weight of oleic acid = 0.28245)

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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:
                                C=N
Molecular formula:  CjH-Cl-N
Molecular weight:   171.9
Melting point:      145-146C for pure compound; the technical product
                    is about 95% pure and has a m.p. 139 to 1A6C
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 20C); 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

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                                                         Dichlooenil 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)

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                                                        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 ja to 13.8 ji) .
         Determine the absorbance of standard and sample using the
     peak at 806.5 cm   (12.A p) and baseline from 819.7 cm   to
     787.4 cm'1 (12.2 >i to 12.7 p).
     Calculation;
         From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent dichlobenil as
     follows:

      _ (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"1 to 720 cm    (11.5 ji to 13.9 >i)
     analytical peak:  780.0 cm'1 (12.82 ji)
     baseline:         819.7 cm"1 to 740.7 cm"1  (12.2 ^ 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.

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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:
Molecular formula:  C.-H.C1_0
                     10 4  22
Molecular weight:   227.1
Melting point:      193C (slowly sublimes above 32C)
Physical state and color:  yellow crystals or leaflets
Solubility:  practically insoluble in water (0.1 ppm at 25C);
             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

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                                 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
                          *
     2.  Mechanical shaker
     3.  Centrifuge or filtration apparatus
     4.  Water bath  40C
                                   *
     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 rag/ml)

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                            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 40C 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:
           i
    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 ^ to 8.16 >i) .
    Determine the absorbance of standard and sample using the
peak at 1275 cm"  (7.84 ji) and basepoint at 1300 cm   (7.69 p) .

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                                 A                       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.

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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:      53C
Boiling point:      173.4C
Physical state, color, and odor:  colorless crystals with a character-
                                  istic penetrating odor.
Solubility:  about 80 ppm in water at 25C; 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 disulflde, pesticide or spectro grade
     3.  Sodium sulfate, anhydrous, granular

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                                                  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
     A.  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 mg/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 ;i to 13.5 ji).
         Determine the absorbance of standard and sample using the peak
     at 816 cm'1 (12.25 >i) and baseline from 855 cm'1 to 794 cm'1 (11.7
     to 12.6  i).

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                                                     p-Dichlorobenzene EPA-1
                                                     (Tentative)
     Calculation;
         From the above absorbances and using the standard and sample
     concentrations, calculate the percent p-dichlorobenzene as
     follows:

      3 (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.

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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.Cl-
                     64  2
Molecular weight:   147.01
                    53C
                    173. 4C
Melting point:
Boiling point:
Physical state, color, and odor:  colorless crystals with a character-
                                  istic penetrating odor
Solubility:  about 80 ppm in water at 25C; 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

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                                 2                  p-Dichlorpbenzene 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 ul
     4.  Usual laboratory glassware

Operating Conditions for TCD;
     Column temperature:     117C
     Injection temperature:  1AOC
     Detector temperature:   140C
     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.

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                                                     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.

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                                                    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

          (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)(100)
         (wt. sample)(pk. ht. or area DDVP)(RF)
Method submitted by Stelios Gerazounis, EPA Region II, New York, N. Y.

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October 1975
Dieloran 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:
                                NH2
Molecular formula:  C,H.C1_N_00
                     64  222
Molecular weight:   207
Melting point:      192 to 194C
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 20C
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

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                                 2                       Dieloran 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

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                            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)

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 1250 cm   to
1042 cm   (8 p. to 9.6 ;i) .
    Determine the absorbance of standard and sample using the
peak at 1147 cm'1 (8.72 ji) and baseline from 1183 cm"1 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:

 _ (abs. sample)(cone. std in mg/ml)(% purity std)
    (abs. std)(cone, sample in mg/ml)

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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:
  CH3
                                                 N02
            N02
       h=0,l,-2  (I )
                                                       0
                                                    0-C-CH=CH-
 (CHZ)*,,	/  ~  0                N02-
HC-	('      Vo-C-CH=CH-CH3
                                    CH3-(CH2)SCH-(CH2VCH3
                                                            (II)
Dinocap is a registered acaricide and  fungicide.

Molecular formula:  C,0H0/N00,
                    lo /4 2. b
Molecular weight:   364
Boiling point:      138 to 140C 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

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                                 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 K^O^.)
     5.  Potassium sulfate, reagent grade  (see above)
     6.  Sodium or potassium sulfide, reagent grade
     7.  Granulated zinc, reagent grade

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                                 3                           Dinocap EPA-1

     8.  Kje'ldahl 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.

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                                                    Dinocap EPA-1
Reduction of NO  Group:
    Weigh a portion of sample equivalent to 0.3-0.4 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.

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                            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 	> NaJZtiCU + 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.

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                            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 H2S04)(N of H2S04)-(ml NaOH)(N of NaOH)](.01401)(100)
                            (grams of sample)


The % dinocap is found by dividing the percent nitrogen by the
percent nitrogen in dinocap.

,  ,.          % nitrogen in sample
% dinocap  =  	f7	c
                     D .D


Technical dinocap contains from 6.6% to 7.2% nitrogen according
to information received from the Rohm and Haas Company, March 1974.

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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 Innethylheptyl,  1-ethylhexyl, and
1-propylpentyl Isomers.  The chemical structures are:
                                                   N02
                   0
                0-C-CH=CH-CH3
   0
   II
0-C-CH=CH-CH3
                                            * = 0,1, or 2  (H)
Dinocap is a registered acaricide and fungicide.

Molecular formula:   C. -.H^-N^O..
                     lO /4 i D
Molecular weight:    364
Boiling point:       138 to 140C 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

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                                 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 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 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/tnl)

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                            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,   (filial cone 10 mg dinocap/ml)
    (There may be interference from the emulsifier in the
     sample; if so, another procedure must be used.)

Pet ermina t ion;
    With carbon disulfide in the reference cell, and using the
optimum quantitative analytical settings, scan both the standard
                       -1           -1
and sample from 1430 cm   to 1250 cm   (7.0 u to 8.0 ji).
    Determine  the absorbance of standard and sample using the peak
at 1340 cm"1 (7.46 ji) and baseline from 1385 cm"1 to 1310 cm~
(7.22 u to 7.63 p).

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,

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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:
                                        CH-CH2-CH3
                                        CH3
Molecular formula:  C QH  N 0
Molecular weight:
Melting point:
240.2
see below
Physical state, color, and odor:   pure  compound - yellow crystals
                    mp 38 to 42C;  technical compound - orange-brown
                    liquid of 95  to 98% purity and mp 30 to 40C;
                    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 177C
Other names: Premerge (Dow),  dinosebe  (France), Bansanite, Chemox,
             Gebutox, DNBP, Dinitro, DN289,  Kiloseb, Nitropone, Sinox

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                                 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, 40C, and a stream of dry air
                                   A
     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 rag/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 ji).

     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 Commonvjealth 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 e'ther as extraction
solvent.

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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
                             N02
Molecular formula:  cioHi2N25
Molecular weight:
Melting point:
240.2
see below
Physical state, color, and odor:  pure compound - yellow crystals
                    mp 38 to 42C; technical compound - orange-brown
                    liquid of 95 to 98% purity and mp 30 to  40C;
                    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 177C

Other names: Premerge (Dow), dinosebe (France), Bansanite, Chemox,
             Gebutox, DNBP, Dinitro, DN289, Kiloseb, Nitropone, Sinox

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                                 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:     220C

         Injection temperature:  250C

         Detector temperature:   250C

         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 mg/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 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.

-------
                                                           Dinoseb EPA-2
                                                           (Tentative)
     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)
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.

-------
November 1975
Diphacinone EPA-1
              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:
                                    0
                                    II
                                    c-
Molecular formula:  C0_H,,00
                     23 16 3
Molecular weight:   340.A
Melting point:      1A5C
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 nrn.

-------
                                 2                      Diphacinone EPA-1
Reagents:
     1.  Diphacinone standard of known % purity
     2.  Sodium pyrophosphate, 1% solution - dissolve 5 grams
         Na,P207.10 H_0 in water and make to 500 ml.
     3.  Ethyl ether, ACS
     4.  Petroleum ether - extract 200 ml with three 20 ml portions
         of 1% sodium pyrophosphate solution.

Equipment:
     1.  Ultraviolet spectrophotometer, double beam ratio recording
         with matched 1 cm silica cells
     2.  Soxhlet extraction apparatus
     3.  Centrifuge
     4.  Aspirator or suction device with fine tip glass tube
     5.  Usual laboratory glassware

Procedure:
     Preparation of Standard;
         Weigh 0.08 gram diphacinone standard into a 100 ml volumetric
     flask, dissolve, make to volume with 1% sodium pyrophosphate
     solution, and mix thoroughly.  Pipette 10 ml into a second 100 ml
     volumetric flask, make to volume with the pyrophosphate solution,
     and mix well.  Pipette 5 ml into a third 100 ml volumetric flask
     and make to volume,  (final cone 4 jig/ml)

     Preparation of Sample:
         Weigh a portion of sample equivalent to 0.001 gram diphacinone
     (20 grams for 0.005% product) into a Soxhlet thimble, plug with
     cotton or glass wool, and extract with ethyl ether for 4 hours.
     Evaporate the extract to less than 50 ml on a steam bath, transfer
     to a 50 ml volumetric flask, and make to volume with ethyl ether.

-------
                            3                      Diphacinone EPA-1

    Pipette 2 ml of the ether solution into a glass-stoppered test
tube (approx. 15 ram x 150 mm).  Add 10 ml 1% sodium pyrophosphate
solution by pipette.  Shake vigorously for 2 minutes and centrifuge
until the aqueous layer is clear.  Draw off the ether layer using
an aspirator fitted with a glass tube drawn into a fine tip.  Add
2 ml ethyl ether, shake vigorously, centrifuge, and draw off ether
layer.  Repeat with another 2 ml ethyl ether.  Finally, repeat
twice with 2 ml portions of petroleum ether.  Centrifuge a few
minutes with the stopper off  the tube to completely evaporate any
residual ether.  (final cone  4 jig diphacinone/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 286 nm  with 1% sodium 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 286 nm and
calculate the percent diphacinone as follows:

   (abs. sample)(cone, std in jug/ml)(% purity std)
    (abs. std) (cone, sample in jig/ml)

or, using dilution factors, as follov/s:

    (abs. sample)(wt. std)(purity std)(1/100)(10/100)(5/100)(100)
/0 = (abs. std)(wt. sample) (1/50) (2/10)

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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-CH2-0       S
                      ^>PS-CH2-CH2-S-CH2-CH3
      CH3-CH2-CT
Molecular formula:   CgH -0-PS
Molecular weight:    274.2
Boiling point:       62C 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.      r
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)
     A.  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 yu)
     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 u to 17.0 ja).  For a qualitative
     comparison,run a full scan.

     Calculation;
                                                                -1
         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)
     / Disulroton = -r-	~r\~7	T	*	/ -i \	
                    (abs. std)(cone, sample in mg/ml)
Method submitted by Dean Hill, EPA Region IX, San Francisco, Calif.

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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:
    CH3-CH20
                           II
                           P-S-CH2-CH2S-CH2CH3
Molecular formula:  C_H,_O.PS,
                     o iy   J
Molecular weight:   274.2
Boiling point:      62C 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
     A.  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 ionizatlon detector (FID)
     2.  Column:  4' 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 jil
     A.  Mechanical shaker
     5.  Centrifuge or filtration equipment
     6.  Usual laboratory glassware

Operating Conditions for FID;
     Column temperature:     190C
     Injection temperature:  2AOC
     Detector temperature:   2AOC
     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 mg
     disulfoton and 2.5 mg 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 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 elutlon 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:

     RF = (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:

      _ (wt. alachlprH% purity alachlor) (pk. ht. or area disulfoton) (100)
         (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,

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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:  CgH10C12N2
Molecular weight:   233.1
Melting point:      158 to 159C
Physical state, color, and odor:  Odorless, white, crystalline solid
Solubility:  42 ppm in water at 25C; slightly soluble in hydrocarbons,
             about 5.3% in acetone at 27C
Stability:   decomposes at 180-190C; 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-dichloroanlline, 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 (SI) 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 200C
     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.

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                                                         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 175C  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:

     % p (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 Chromatography
     Diuron is the common name for 3-(3,4-dichlorophenyl)-l,l-dimethyl-
urea, a registered herbicide having the chemical structure:
Molecular formula:  C.H10C12N2
Molecular weight:   233.1
Melting point:      158 to 159C
Physical state, color, and odor:  Odorless, white, crystalline solid
Solubility:  42 ppm in water at 25C; slightly soluble in hydrocarbons,
             about 5.3% in acetone at 27C
Stability:   decomposes at 180-190C; 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


                                           o
     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 rag 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:


      = (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 QC1 N20
Molecular weight:   233.1
Melting point:      158 to 159C
Physical state, color, and odor:  Odorless, white, crystalline solid
Solubility:  42 ppm in water at 25C; slightly soluble in hydrocarbons,
             about 5.3% in acetone at 27C
Stability:   decomposes at 180-190C; 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 run to 200 nm 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 ug/ml.

     Calculation;
         From the above absorbances and using the standard and sample
     concentrations, calculate the percent diuron as follows:

     7 - (abs. sample)(cone, std in pg/ml)(% purity std)
         (abs. std) (cone, sample in jig/ml)
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  C1_N 0
Molecular weight:   233.1
Melting point:      158 to 159C
Physical state, color, and odor:  odorless, white, crystalline solid
Solubility:  42 ppm in water at 25C; slightly soluble in hydrocarbons,
             about 5.3% in acetone at 27C
Stability:   decomposes at 180-190C; 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-A
                                                       (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 small 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   (6.67 ji to 7.7 ji) .
         Determine the absorbance of standard and sample using the peak
     at 1353 cm   (7.39 p) and baseline from 1399 cm"  to 1316 cm"
     (7.15 ji 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)(cone, std 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 liexachlorohexahydro-
methano-2,4,3-benzodioxathiepin-3-oxide, a registered pesticide
having the chemical structure:
                                                      S=0
              Cl
Molecular formula:
Molecular weight:
Melting point:
C9H6C163S
406.9
(see below)
Physical state, color, and odor:  endosulfan is an odorless white
                    crystalline solid mixture of two isomers with
                    mp's of 106C and 212C; the technical product
                    is a brownish crystalline solid, mp 70-100C,
                    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:  Thlodan (Farwerke Hoechst),  Beqsit, 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, 14-4
     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 S02.

-------
                                                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 titration on 40 ml of standard 0.1N iodine solution
using 175 ml distilled water and 1 ml 1 -f 4 sulfuric acid.

Calculation;
    The molecular weight of endosulfan is 406.95 and the milli-
equivalent weight is 0.2305.

Net ml Na.S-O. used = ml Na_S 0  for blank - ml Na-S.O. for sample

               (net ml Na_S,0 )(N of Na.S 0 )(.2035)(100)
     Z*   * f                * j           j
  endosulfan  -.-	~. v/en/-inn	~T~~\	
               (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
                                                      S0
Molecular formula:  CftH,Cl,0_S
                     96  63
Molecular weight:
Melting point:
406.9
(see below)
Physical state, color, and odor:  endosulfan is an odorless white
                    crystalline solid mixture of two isotners with
                    mp's of 106C and 212C; the technical product
                    is a brownish crystalline solid, mp 70-100C,
                    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, Cycledan,
              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 sulfate 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 endosulfan/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"1 (8 p to 9 p).
         Determine the absorbance of standard and sample using the
     peak at 1192 cm   (8.39 p) and baseline from 1205 cm   to 1176 cm
     (8.3 p to 8.5 p).

     Calculation;
         From the above absorbances and using the standard and sample
     concentrations, calculate the percent endosulfan as follows:

      m (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 Chromatbgraphy
                    (TCD - 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
                                        CH2
                                        CH2  0'
                                                     >S=0
                                 H
Molecular formula:  C.H.C1,0,S
                     70  O J
Molecular weight:   A06.9
Melting point:      (see below)
Physical state, color, and odor:  endosulfan is an odorless white
                    crystalline solid mixture of two isomers with
                    rap's of 106C and 212C; the technical product
                    is a brownish crystalline solid, mp 70-100C,
                    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, Thlraul, 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 jil
     4.  Usual laboratory glassware

Operating Conditions for TCP:
     Column temperature:     230C
     Injection temperature:  260C
     Detector temperature:   260C
     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 4 rag endosulfan and 2 mg 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 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 HHDN, then endosulfan.
         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 endosulfan and dieldrin
     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:


        a (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:


     % m (wt. HHDN)(% purity HHDN)(pk. ht. or area endosulfan)(100)
         (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
Endosulfan EPA-4
(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:
                                         CH20,
                                         CH2
                                                       .5=0
Molecular formula:  C.H.-Cl.O-S
                     y o  o 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 106C and 212C; the technical product is a
                    brownish crystalline solid, mp 70-100C, 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 5462, 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 pi
      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 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 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)(% purity HEOD)(pk. ht. or area endosulfan)	
           (wt. endosulfan)(% purity endosulfan)(pk. 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 endosulfan) (100)
       = (wt. sample)(pk. ht. or area HEOD)(RF)
      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
CHCOOH
                                     CHCOOH
Molecular formula:  C0HinOc
                     o 1U D
Molecular weight:   186.2
Melting point:      144C (some decomposition, see below)
Physical state, color, and odor:  white,  odorless, crystalline solid
Solubility:  solubility in grams per  100  ml at 25C is:  10 in water,
             7 in acetone, 0.1 in benzene, 7.6 in dioxane, 28 in methanol
                                                                        I
Stability:   stable to light;  stable  to about 90C, 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.)

-------
                                 3                     Endothall EPA-1


     Neutralize carefully with 0.1N sulfuric acid to just colorless
with phenolphthalein.  Evaporate and ash at approx. 525C.  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 sulfuric acid solution and boil 20 minutes
to remove carbon dioxide.  Cool, and titrate with 0.1N sodium hydroxide
solution to the phenolphthalein endpoint.
     Calculate the endothall as follows:

     % 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

     % endothall acid X 1.236    % endothall disodium salt

-------
December 1975
                     Endothall EPA-2
                     (Tentative)
                    Determination of Endothall
                by Gas-Liquid Chromatography (FID)

     Endothall is the accepted common name for  7-oxabicyclo  (2.2.1)
heptane-2,3-dicarboxylic acid, a registered herbicide  having the
chemical structure:
                                            COOH
               Ho-C
CHCOOH
Molecular formula:  C0H,-CL
                     O 1U D
Molecular weight:   186.2
Melting point:      144C (some decomposition,  see below)
Physical state, color, and odor:  white, odorless, crystalline solid
Solubility:  solubility in grams per 100 ml at  25C 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 90C, 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 I/A" O.D. glass, packed with 3% SE-30 on
                  60/80 Chromosorb W AW DMCS (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:     130C

     Injection temperature:  180C

     Detector temperature:   180C

     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

     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 fig endothall/fil)
         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 jig endothall/jil)

     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:

      = (ok. 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 EPA1s 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
                          ||           ,CH2CH2CH3
  CH3	CH2	S	C	N <^
                                      XH2 CH2CH3
Molecular formula:   C_H  NOS
Molecular weight:   189.3
Boiling point:      127C at  20 mm Hg  (235C by extrapolation)
Physical state, color,  and odor:  Light yellow-colored liquid with
                                 an amine odor
Solubility:  365 ppm in water at  20C; 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:     130C
     Injection temperature:  225C
     Detector temperature:   150C
     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 EPTC
     and 10 mg ethyl hexanediol/ml)

-------
                                 3                       EPTC   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 mg 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)(100)
         (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 EPIC by        :   .;
               High  Pressure Liquid Chromatography
     EPIC is the common name for S-ethyl dipropylthibcarbamate, a
registered herbicide having the chemical structure:


                          ?        /CH2-CH2CH3
  CH3CH2SC	N<^
                                     XH2CH2CH3

Molecular formula:  CgH gNOS
            i
Molecular weight:   189.3
Boiling point:      127C  at 20 mm Hg (235C  by extrapolation)
Physical state,  color, and odor:  Light yellow-colored liquid with
                                 an amine odor
Solubility:        365 ppm in water at 20C; miscible with acetone,
                   benzene, ethanol, isopropanol, kerosene, methanol,
                   methyl isobutyl ketone, toluene,  and xylene
Stability:         stable, non-corrosive

Other names:       Eptam  (Stauffer), Eradicane,  Knoxweed

                                -'.  \   '   i .
Reagents;
     1.  EPTC standard of  known % purity
     2.  Chloroform
     3.  Dichloromethane
     4.  Hexahe
     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 u (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 EPTC/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:


     ~ = (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

Toshihiko 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)

     EPIC is the common name for S-ethyl dipropylthiocarbamate, a
registered herbicide having the chemical structure:
                           0
                           II        ^CH2CH2CH3
   CH3 CH2 S C	N
                                             !CH2CH3
Molecular formula:   C H  NOS
Molecular weight:   189.3
Boiling point:      127C at  20 mm Hg  (235C by extrapolation)
Physical state, color, and odor:  Light yellow-colored liquid with
                                 an amine odor
Solubility:  365 ppm in water at  20C; 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.  EPIC 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.  Methanol, pesticide or spectro grade
      :, , .; ,..,. ,-j-fg -}s* j 'i   ' :
     6.  Internal Standard solution - weigh 0.25 gram butylate into
         a 50 ml volumetric
-------
                                 3                          EPIC   EPA-3
Procedure;
     Preparation of Standard;
         Weigh 0.08 gram EPIC 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 EPIC
     and 5 rag 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)

     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 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                       EPTC   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 EPTC)
        ~ (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 EPTC)(100)
      ~ (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-4
                                                       (Tentative)
                    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:
                           ||           CH2CH2CH3
   CH3 CH2SC	N <^
                                                CH2CH3
Molecular formula:  C H  NOS
Molecular weight:   189.3
Boiling point:      127C at 20 mm Hg (235C by extrapolation)
Physical state,  color, and odor:  Light yellow-colored liquid with
                                 an amine odor
Solubility:  365 ppm in water at 20C; 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                     EPTC   EPA-4
                                                       (Tentative)
Reagents;
     1.  EPTC 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
         + 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 jjl
     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 EPIC 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
     EPIC and 4 mg vernolate/ml)

     Preparation of Sample:
         Weigh a portion of sample equivalent to 0.05 gram EPIC
     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 arid 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 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.

-------
                                                        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:


     R_ _ (wt. vernolate)(% purity vernolate)(pk. ht. or area EPIC)
          (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) (100)
         (wt. sample)(pk. ht. or area vernolate)(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.

-------
October 1975                                          EPIC   EPA-5
                                                      (Tentative)
                     Determination of EPIC by
                    GasLiquid Chromatography
                    (TCD -  Internal Standard)

     EPIC is the common name  for S-ethyl dipropylthiocarbamate,  a
registered herbicide having the chemical structure:
                           0
                           ||            CH2CH2CH3
  CH3	CH2	SC	N
                                              CH2
Molecular formula:  C H  NOS
Molecular weight:   189.3
Boiling point:      127C at  20 mm Hg  (235C by extrapolation)
Physical state, color, and odor:  Light yellow-colored liquid with
                                 an amine odor
Solubility:  365 ppra in water at  20C; 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 % 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:     160C
     Injection temperature:  200C
     Detector temperature:   200C
     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 EPTC 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
     EPTC and 10 mg vernolate/ml)

     Preparation of Sample:
          Weigh a portion of sample equivalent to 0.08 gram EPTC
     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 /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 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)
          (wt. EPTC)(% purity EPTC)(pk. ht. or area vernolate)
          Determine the percent EPIC for each injection of the

     sample-internal standard solution as follows and calculate the

     average:


     , _ (wt. vernolate)(% purity vernolate)(pk. ht. or area EPIC)(100)
         (wt. sample)(pk. ht. or area vernolate)(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.
                                                                        Q

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  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:
CH3CH20   S                       S     0CH2CH3
                   \SCH2S  P<^
CH3CH20                               \,_CH2 CH3
  Molecular formula:  C.H-.O.P.S.
                       9 22  4  2 4
  Molecular weight:   384.48
  Boiling point:      164 to 165C at 0.3 mm Hg;  solidifies at -12 to
                      -15C
  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 grains 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 fi 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 p 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 mg/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"1 (10.19 ;i)  basepoint

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  October 1975                                            Ethion EPA-2
                                                          (Tentative)
                     Determination of Ethion by
                   Gas-Liquid Chromatography (TGD)

       Ethion is the  accepted common name for 0,0,0', O'-tetraethyl
  S,S'-raethylene bisphosphorodithioate, a registered  insecticide having
  the chemical structure:
CH3CH20   S                       S     0CH2CH3
                   \PS CH2 S	P<^
CH3CH20                               \_CH2CH3
  Molecular formula:   CgH-.O P S,
  Molecular weight:    384.48
  Boiling point:       164  to 165C 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 chroraatograph with thermal conductivity detector (TCD)

     2.  Column:  5' x 1/4" glass column packed with 10% QF-1 on
         Chromosorbi 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:     210C

         Injection temperature:  240C

         Detector temperature:   240C

         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)

-------
                                                          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:

     / - (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 nematoclde and soil insecticide having the
chemical structure:
                                    -S	CH2 CH2CH3
                            M ^
    CH3CH20-
                                    S	CH2CH2CH3
Molecular formula:   C-H.-O-PS,,
                     O  17  /  L
Molecular weight:    2A2.3
Melting point:      86  to  91C 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  100C;
             hydrolyzed in basic media moderately fast at  25C and
             rapidly at 100C; thermal stability is good for 8 hours
             at 150C

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 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.  Filtration apparatus or centrifuge
     4.  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 Erlernneyer 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-1
                                                       (Tentative)

         For emulsifiable 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.)

     De terminat ion t
         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 i. to 11.1 i).
         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 u to 10.3 p.).

     Calculation!
         From the above absorbances and using the standard and sample
     concentrations, calculate the percent ethoprop as follows:

         %  (abs. sample) (cone, std inmg/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.

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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 nematocide and soil insecticide having  the
chemical structure:
   CH3 CH2 0
                                    S(HzCH2CH3
                                      CH2CH2CH3
Molecular formula:   C0Hn_O.PS-
                     o  iy  L  i
Molecular weight:    242.3
Melting point:       86  to  91C 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 100C;
             hydrolyzed in basic media moderately fast at 25C and
             rapidly at 100C; thermal stability is good  for 8 hours
             at  150C

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:     200C
     Injection temperature:  225C
     Detector temperature:   225C
     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 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 8 mg ethoprop and 10 mg 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:


        m (wt. diazinon)(% purity diazihon)(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:


     . ^ (wt. diazinon)(% purity diazinon)(pk. ht. or area ethoprop)(100)
         (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
    CH3CH20	P:
                               \
                                    'S  CH2CH2 CH3
Molecular formula:   C H  0 PS
                     8 1" 2  L
Molecular weight:    242.3
Melting point:      86 to 91C 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 100C;
             hydrolyzed in basic media moderately fast at 25C and
             rapidly at 100C; thermal stability is good  for 8 hours
             at 150C

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
     A.  Mechanical shaker
     5.  Centrifuge or filtration equipment
     6. . Usual laboratory glassware

Operating Conditions for FID:
     Column temperature:     180C
     Injection temperature:  230C
     Detector temperature:   230C
     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
reproducib11ity.

-------
                                                      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 diazinoh/ml)

     Preparation of Sampler
         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 t
         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:

         (vt. 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 diazinon)(pk. ht. or area ethoprop)(100)
         (wt. sample)(pk. ht. or area diazinon)(RF)
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 solvent1or 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-hexanedibl, a registered insect repellent having the chemical
structure:
                    CH2CHj
            CH2CH	CH	CH2CH2CH3
            OH             OH
Molecular formula:  C0H100.
                     o lo i
Molecular weight:   146.2
Boiling point:      244C;  the technical product has a distillation
                    range of 240 to  250C
Physical state, color, and odor:  colorless liquid; the technical product
                    has a faint odor of witch hazel
Solubility:  slightly soluble in water; tniscible with alcohol, chloroform,
             ether; will not dissolve nylon, rayon
Stability:   stable under normal conditions; both hydroxyl groups can be
             esterifled, the secondary group with difficulty; it is
             without chemical or solvent action on clothing and most
             plastics

Other names: ethbhexadiol (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 hexanedlol

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:
                    CH2CH3
            CH2  CHCH	CH2 CHzCHj
            OH            OH
Molecular formula:   C0H,00_
                     o  lo  i.
Molecular weight:    146.2
Boiling point:       244C; the technical product has a distillation
                    range  of 240 to 250C
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-2
                                                     (Tentative)
Reagents;
     1.  2-Ethyl-l,3-hexanedlol.of known % purity
     2.  o-Dichlorobenzene, commercial grade or better
     3.  Isopropanol, pesticide or spectre 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 ul
     4.  Usual laboratory glassware

Operating Conditions for TCD;
     Column temperature:     120C
     Injection temperature:  150C
     Detector temperature:   150C
     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.

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                                                     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)

     Determination:
         Inject 1 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 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:

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                                                      Ethyl Hexanediol EPA-2
                                                      (Tentative)
         I.S.  Internal Standard = o-dichlorobenzene


          (wt. I.S.)(% purity I.S.)(pk. ht. or area ethyl 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)(100)
      ~ (wt. sample)(pk. ht. or area I.S.)(RF)
     Note:  a 1/4" column can be used at 130C 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.

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November 1975
Fluometuron EPA-1
                   Determination of Fluometuron
                     by Infrared 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  H  F N.O
Molecular weight:   232.2
Melting point:      163 to 164.5C (The technical product is about
                    96% pure and has a m.p. of about 155C)
Physical state, color, and odor:  odorless, white, crystalline solid
Solubility:  90 ppm in water at 25C; 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

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                                 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  Na.SO,.  Pipette 50 ml
     chloroform into the flask.  Shake the sample on a mechanical shaker
     for one hour.  Transfer a portion of the GHC1. 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,

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                                 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 mg 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 /i).
         Determine the absorbance of standard and sample using the
     peak at 1335 cm"1 (7.49 ft) and baseline from 1355 era   to 1300 cm
     (7.38 ji to 7.69 ji).

     Calculation:
         From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent fluometuron as
     follows:

     v a (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.

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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:
                                 NSCCl
Molecular formula:  CgH.Cl.NO-S
Molecular weight:   296.6
Melting point:      177C
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

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                                 2                        Folpet EPA-1
Equipment;
     1.  Infrared spectrophotoraeter, double beam ratio recording with
         matched 0.1 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,
  1       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/ml)

     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)

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                                 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 (5.26jj to 6.1;u).
         Determine the absorbance of standard and sample using the
     peak at 1755 cm"  (5.70 ji) and basepoint at 1850 cm"  (5.41 ,u).
     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 rag/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.

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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
                                 H
                                                 H
Molecular formula:  Clf.H Cl_
Molecular weight:
Melting point:
373.5
95 to 96C
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 74C
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
                                                             H

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                                 2                     Heptachlor EPA-1
Other names:  Velsicol 104, E3314 (Velsicol Chem. Corp.); Drinox;
              Heptatnul; H-34; l,4,5,6,7,8,8-heptachloro-3a,4,7,7a-
              tetrahydro-4,7-methanoindene

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. 20-40C), 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 rag/ml)

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                            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
instrument being used, scan both the standard and sample from 700 cm
to 625 cm"  (14.3 p to 16.0 p).
    Determine the absorbance of standard and sample using the peak
at 658 cm   (15.2 ji) and baseline from 673 cm~  to 637 cm"  (14.85/j
to 15.7  i) .

-------
                                                     Heptachlor EPA-1
Calculation:

    From the above absorbances and using the standard and sample

concentrations, calculate the percent heptachlbr as follows:


>/ - (abs. sample)(cone, std in mg/ml) (% purity std)
    (abs. std)(cone, sample in mg/ml)

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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-COH
Molecular formula:   C   H  NO-
Molecular weight:    203.2
Melting point:       124C
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 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 0.1 gram indolebutyric acid into a 100 ml volumetric
     flask, dissolve in, and make to volume with 0.5% 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 fig 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 Erlerimeyer 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.

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                                              Indolebutyric acid EPA-1
UV Determinant ion:
    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.
                  i
Calculation;
    From the above absorbances and using the standard and sample
concentrations, calculate the percent indolebutyric acid as
follows:

 _ (abs. sample)(cone. std in ^ig/ml)(% purity std)
    (abs. std) (cone, sample in

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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
                                           N  C CH3
                                NCN\
                                /.    !     V
Molecular formula:  C1/H0.N,00
                     14 21 3 3
Molecular weight:   279.A
Melting point:      176 to 176.5C
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

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                                 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)

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                                 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
     1650 cm   (5.2 p to 6.0 ji).
         Determine the absorbance of standard and sample using the
     peak at 1745 cm   (5.73 ;i) and basepoint at 1840 cm"  (5.43^i).
     Calculation;
         From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent karbutilate as
     follows:

     _, _ (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 ji)
     analytical peak:  as above
                       along she
                       (5/i to 5.56
baseline:         along shoulder from about 2000 cm   to 1800 cm

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July 1975                                                Llnuron EPA-1
                                                          (Tentative)
                   Determination of Linuron by
               High Pressure Liquid Chromatography
     Linuron is the common name for 3-(3,4-dichlorophenyl)-l-methoxy
 -1-methylurea, a registered herbicide having the chemical structure:
                          0-U 0
                           13II
          CH30NCN
Molecular formula:  CQH10C12N22
Molecular weight:   249.1
Melting point:      93 to 94C
Physical state, color, and odor:  odorless, white, crystalline solid
Solubility:  75 ppm in water at 25C; 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.

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                                 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 fi (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/min


     Detector:                  UV at 254 run


     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 0.01 gram linuron standard into a 50 ml volumetric


         flask; dissolve and make to volume with chloroform (final


         cone  0.2 mg/ml).

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                                 j                         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 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 linuron as follows:


     ,. m (pk. ht. or area sample)(vt. 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.

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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
      CHj0N	CN
Molecular formula:  CoHroC12N22
Molecular weight:   249.1
Melting point:      93 to 94C
Physical state, color, and odor:  odorless, white,  crystalline  solid
Solubility:  75 ppm in water at 25C;  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

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                                 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)

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                                                         Linuron EPA-2
     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 1370 cm   to
     1250 cm'1 (7.3 p to 8.0 >i).
         Determine the absorbance of standard and sample using the
     peak at 1290 cm~  (7.75 ;i) 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:

      m (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.

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December 1975
                                                       Malathlon 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
CH2
                         S CH - C - 0 - CH2 CH3
                                         0
                                         C  0  CH  
                                                     CH2  CH3
Molecular formula:   C,nH.Q0>PS_
                     1U 1"  0   L
Molecular weight:    330. A
Melting/boiling point:   m.p.  2.85C, b.p. 156 to 157C 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
             (30-60C)  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.

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                                 2                      Malathion EPA-1
                                                        (Tentative)
Other names:  El 4049 and Cythion (American Cyanamid), mercaptothion
              (So. Africa), carbofos (USSR), Etnmatos, 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 run.
         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 malathion.
     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:  55C
     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 reproducibillty.

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                                                        Malathion 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 ug/pl)

     Preparation of Sample;
         Weigh an amount of sample equivalent to 0.5 gram malathion into
     a glass-stoppered flask or vial, add 100 ml methanol 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 jig malathlon/jil)

     Determination;
         Using a high pressure liquid syringe or sample injection loop,
     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
     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, Beltsvllle, Md.

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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:
                                                0  CH2 CH3
       CH3
                                          0
                                          II
                                 CH2  C - 0  CH2 CH3

Molecular formula:  C1_H100,PS0
                     1U 1? D  i
Molecular weight:   330.4
Melting/boiling point:  m.p. 2.85C,  b.p. ,156  to 157C 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
             raise ible with most organic  solvents; light petroleum oil
             (30-60C) 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 , Fyf anon ,
             Karbofos, Kop-Thion, Kypfos,  Malaspray, Malamar, MLT,
             Zithiol

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                                 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
                                   *
     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 malathion standard into a 10 ml volumetric flask
     and make to volume with carbon disulfide.  Add a small amount of
                                                               I
     anhydrous sodium sulfate to Insure dryness and shake thoroughly.
     (cone 10 mg/ml)

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                                                  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 raalathion/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 ya to 18.0 ji).
    Determine the absorbance of standard and sample using the
peak at 657.9 cm   (15.2 ^i) and basepoint 625 cm   (16.0yj).

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                                                         Malathion EPA-2
     Calculation!
         From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent malathion as
     follows:

      = (abs. sample)(cone. std in mg/ml)(% purity std)
         (abs. std)(cone, sample in mg/ml)

     (A concentration of 1 mg 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.

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November 1975
              Metaldehvde EPA-1
                   Determination of Metaldehyde
                     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:
                                 c
                                 A
                 CH-G H
HC-CH3
                                 CH3
Molecular formula:
Molecular weight:   (44.1)
                          n       .
Melting point:      in scaled tube, 246C;  sublimes at 110 to 120C
                    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 17C); low
             solubility in ethanol (1.8% at 70C) 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

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                                 2                    Metaldehyde EPA-1
Reagents:
     1.  Sulfuric acid, IN solution
     2.  Sodium metabisulfite, 2.5% solution - dissolve 25 grains
         Na~S90  in water and make to one liter.
     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.
     A.  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 100C
     A.  Heating mantle or water bath for 60-70C
     5.  Compressed air
     6.  Steam generator
     7.  Titration apparatus
     8.  Usual laboratory glassware

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                                 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
     60-70C 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 60-70C 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-rbisulfite solution to a 500 ml
     Erleriraeyer flask, add a few ml  starch indicator, titrate the excess
     bisulfite solution by adding about 5 ml of the IN iodine solution,

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                            A                   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 grains 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 =	-.	r-v-
                           (grams sample)
     milliequivalent weight of metaldehyde = 0.02203
     1 ml 0.1000N iodine solution = 0.0022 gram metaldehyde

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November 1975
                                Metaldehyde EPA-2
                                (Tentative)
                   Determination of Metaldehyde
                   by Gas-Liquid Chromatography
                                 ,                        I
                    (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:
                 CH3-
                         0
Molecular formula:  (CH.GHO)
                       3    n
     CH       HC
-CH3
                     0
Molecular weight:
Melting point:
<**.
in sealed tube, 246G;  sublimes  at  110  to  120C
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 17C);  low
             solubility in ethanol (1.8% at 70C)  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

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                                 2                     Metaldehyde EPA-2
                                                       (Tentative)
Reagents:
     1.  Metaldehyde standard of known % purity
     2.  Octyl alcohol standard of known % purity
     3.  Chloroform, pesticide or spectro 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 C AW DMCS (or equivalent column)
     3.  Precision liquid syringe:  10 jjl
     4.  Usual laboratory glassware

Operating Conditions for TCD;
     Column temperature:      90C
     Injection temperature:  140C                              ,
     Detector temperature:   140C
     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.

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                                 3                    Metaldehydc 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)
     IL^JL^Z-^JL^OJIOJL Sample:
         Weigh a portion of sample equivalent to 0.35 gram metaldehyde
     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 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)

     Determinatiori :
         Inject 4-6 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 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.

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                                                     Metaldehyde EPA-2
                                                     (Tentative)
         Determine the RF value for each injection of the standard-
     internal standard solution as follows and calculate the average:

     (vt. octyl alcohol) (% purity 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:
7 = (wt. octyl alcohol) (% purity octyl alcohol) (pk. ht. or area metaldehyde) (100)
    (wt. sample) (pk. ht. or area octyl alcohol) (RF)
Method submitted by Stelios Gerazounis, EPA, Region II, New York, N.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.

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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)
                            n
Molecular weight:
Melting point:
(44.1)
      n
in sealed tube, 246C; sublimes at 110 to 120C
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 17C); low
             solubility in ethanol (1.8% at 70C) 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

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                                 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)

     Preparation of Sample:
         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)

     Determination;
         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'1 (8.0 p to 9.0;i).

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                                                        Metaldehyde EPA-3
                                                        (Tentative)
         Determine the absorbance of standard and sample using the peak
     at 1164 cm"  (8.59 u) and basepoint 1140 cm"  (8.77yu).
     Calculation:
         From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent metaldehyde as
     follows:
     7 = (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.

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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	C	0
                                 H
                 CH3CH      HCCH3
                                 V
                         0	C	0
Molecular formula:  (CH.CHO)
Molecular weight:   (44.1)
Melting point:      in sealed tube, 246C; sublimes at 110 to 120C
                    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 17C); low
             solubility in ethanol (1.8% at 70C) 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, raetacetaldehyde

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                                 2                    Metaldehyde EPA-A
                                                      (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:     120C
     Injection temperature:  160C
     Detector temperature:   160C
     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)

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                                 3                     Metaldehyde EPA-4
                                                       (Tentative)
     Preparation of jaample:
         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)

     Determination:
         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:

      = (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.

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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:
          CH3-S
                       CH3
Molecular formula:  C-.tL.-NCLS
Molecular weight:   225.3
Melting point:      121C
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); mercaptodlmethur,
             metmercapturon, Draza

Reagents;
     1.  Methiocarb standard of known % purity
     2.  Chloroform, pesticide or spectro grade
     3.  Sodium sulfate, anhydrous, granular

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                                                      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
     A.  Cotton or glass wool
     5.  Centrifuge or filtration apparatus
     6.  Rotary evaporator
     7.  Usual laboratory glassware
Procedure:
     Preparation of Standard:
         Weigh 0.07 gram methlocarb 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 methlocarb 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)

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                                                 Methiocarb EPA-1
                                                 (Tentative)
IR Determination t
    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 fi).
    Determine the absorbance of standard and sample using the
peak at 1748 cm"  (5.72 p) and a baseline from 1835 cm"  to
1667 cm"1 (5.45 ji to 6.00
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)

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 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:
CH3 0
                          Cl	C Cl
                                 Cl
Molecular formula:  C.,H.CC1.0.
                     JLO J.J  o 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 89C; technical 70 to 85C
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

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                                  2                   Methoxychlor EPA-1
                                                      (Tentative)
Reagents;
     1.  Methoxychlor, technical standard (minimum 88% p,p* isomer)
     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.  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.
     methoxychlor/ml)

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                                  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 p).
         Determine the absorbance of standard and sample using the
     peak at 795.5 cm'1 (12.57 ji) and basepoint at 772.2 cm'1 (12.95
     Calculation;
         From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent technical methoxy-
     chlor as follows:

      B (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'1  (8.0 ji), 1179 cm'1 (8.48 p), 1042 cm'
 (9.6 ju), or 752 cm'  (13.3 /i) may also be used when interference from
  other ingredients is present;  however, the linearity should be checked.

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 November 1975                                      Methoxychlor  EPA-2
                                                    (Tentative)

                   Determination of Methoxychlor
                    by GasLiquid 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	('          J	C	(/        \)	0  CH3
Molecular formula:  C, ,H, CC1,,0-
                     lo ID  j 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 89C; technical 70 to 85C
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

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                                  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
      4.  Internal Standard solution - weigh 0.2 gram HEOD into a 50 ml
          volumetric flask; dissolve in  and make to volume with acetone.
          (cone 4 rag 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 jil
      4.  Mechanical shaker
      5.  Centrifuge or filtration equipment
      6.  Usual laboratory glassware

Operating Conditions for FID;
      Column temperature:     190C
      Injection temperature:  240C
      Detector temperature:   240C
      Carrier gas:            Nitrogen
      Carrier gas pressure:   40-60 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 rag HEOD/ml)

      Preparation of Sampler
          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:

-------
                                  4                  Methoxychlor EPA-2



      __ _ (wt. HEOD)(% purity HEOD)(pk. ht. or area methoxychlor)	
           (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)(100)
          (wt. sample)(pk. ht. or area HEOD)(RF)



      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.

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November 1975                                      Methyl Parathion EPA-1
                                                   (Tentative)

                Determination of Methyl Parathion
              by High Pressure Liquid Chromatography

     Methyl parathlon 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
Molecular formula:  C0H -NO PS
                     o 10  5
Molecular weight:   263.2
Melting point:      35-36C
Physical state and color:  white crystalline solid; the technical
                    product is a light to dark tan liquid of about
                    80% purity, crystallizing at about 29C.
Solubility:  55-60 ppm in water at 25C; 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 parathlon;
             E601; Folidol M; Fosferno M50; Gearphos; Metron; Partron M;
             Tekwaisa; Wofatox

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                                 2                   Methyl Parathion EPA-1
                                                     (Tentative)
Reagents;
     1.  Methyl parathlon standard of known % purity
     2.  Methanol, pesticide or spectro grade

Equipment;
     1.  High pressure liquid chrornatograph 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 4- 80% water
         Column temperature:  50-55C
         Chart speed:         5 min/inch or equivalent
         Flow rate:           0.5 to 1.5 ral/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.

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                                                    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:

     ~ a (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.

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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 0-p-nitrophenyl phosphorothioate, a regis-
tered insecticide having the chemical structure:
Molecular formula:  C_H,nNO_PS
                     o ID  j
Molecular weight:   263.2
Melting point:      35-36C
Physical state and color:  white crystalline solid; the technical
                    product is a light to dark tan liquid of about
                    80% purity, crystallizing at about 29C.
Solubility:  55-60 ppm in water at 25C; 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

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                                 2                  Methyl Parathlon 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 emulsifiable 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)

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                            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   (7.4 i 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"
(7.85 u to 8.35 ;i) .

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 mg/ml)

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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-methyl,  ISO
and BSI) for 0,0-ditnethyl 0-p-nitrophenyl phosphorothioate,  a regis-
tered insecticide having the chemical structure:
     GH3	0
    CH3'
Molecular formula:  C0H,_NO.PS
                     o 1U  j
Molecular weight:   263.2
Melting point:      35-36C
Physical state and color:  white crystalline .solid; the technical
                    product is a light to dark tan liquid of about
                    80% purity, crystallizing at about 29C.
Solubility:  55-60 ppra in water at 25C; 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

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                                 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 run 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/ral.

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

-------
                            A                 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 ug
    methyl parathion/ml or approx. 3 jig p-nitrophenol/ml .

Calculation;
    Using the absorbance due to the free p-nitrophenol (FPNP) ,
calculate the percent present as follows:

    7   (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:
    v   (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 - % free p-nitrophenol

    The % methyl parathion is then found by dividing this %
p-nitrophenol by .5285 or multiplying by 1.892.
  Methyl Parathion =       ooc          r (1-892) (% p-nitrophenol)
                        V .
Methyl parathion   =  52.85% p-nitrophenol
% methyl parathion   % p-nitrophenol X 1.892

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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:
     CH3	0
     CH30
Molecular formula:  C_H,-NOCPS
                     o 1U  ;>
Molecular weight:   263.2
Melting point:      35-36C
Physical state and color:  white crystalline solid; the technical
                    product is a light to dark tan liquid of about
                    80% purity, crystallizing at about 29C.
Solubility:  55-60 ppm in water at 25C; 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

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                                 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:  6* 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 Chrora 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:     190C
     Injection temperature:  215C
     Detector temperature:   260C
     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 Sample:
         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.

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                                 ,4                   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)(100)
      0 = (wt. sample)(pk. ht. or area DDE)(RF)
Method submitted by Mississippi State Chemical Laboratory, Box CR,
Mississippi State, Mississippi 39762.

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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 0,0-dimethyl 0-p-nitrophenyl phosphorothioate, a regis-
tered insecticide having the chemical structure:
                                                         N02
Molecular formula:  C0H,-NOCPS
                     o 1U  j
Molecular weight:   263.2
Melting point:      35-36C
Physical state and color:  white crystalline solid; the technical
                    product is a light to dark tan liquid of about
                    80% purity, crystallizing at about 29C.
Solubility:  55-60 ppm in water at 25C; 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

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                                 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:  61 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:     175C
     Injection temperature:  250C
     Detector temperature:   2508C
     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.

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                                 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 mg 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:

     % = (wt. HEOD)(% purity HEOD)(pk. ht. or area methyl parathion)(100)
         (wt. sample)(pk. ht. or area HEOD)(RF)
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
              Chroraosorb W HP
     Column temp:  180
     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.

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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:
                                                     CH3
Molecular formula:  C H  BrN 0

Molecular weight:   259

Melting point:      95.5 to 96C

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

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                                                    Metobromuron EPA-1
                                                    (Tentative)
Equipment;
     1.  Infrared spectrophotometer, double beam ratio recording
         with matched 0.5 mm Kfir 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
     sample from 1430 cm"  to 1250 cm"   (7.0 ji to 8.0 ji).
         Determine the absorbance of standard and sample using the
     peak at 1387 cm"  (7.21 p) and basepoint 1351 cm"  (7.40 p).

-------
                                                       Metobromuron EPA-1
                                                       (Tentative)
         An alternate peak at 1305 cm   (7.66 p) with the same
     basepolnt 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:

      _ (aba, 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 11, New York, N. Y. submitted a similar
method using:
     scan range:       2000 cm"  to 1430 cm'   (5.0 ji to 7.0^i)
     analytical peak:  1683.5 cm   (5.94 ji)
     basepoint:        1818 cm   (5.5 ^i)

     The absorption curve is linear for 2-16 mg/ml.
Comments on these analytical bands  (or others) are most welcome.

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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:
                                                    CH3
Molecular formula:  C H  BrN 0
Molecular weight:   259
Melting point:      95.5 to 96C
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.  Coltunn:  21 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:     165C
     Injection temperature:  200C
     Detector temperature:   200C
     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)

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                                                      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/ml)

     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:

     7 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.

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November 1975                                        Metobromuron EPA-3
                                                     (Tentative)
                  Determination of Metobromuron
                   by Gas-Liquid Ghromatography
                    (TCD - Internal Standard)

     Metobromuron is the accepted common name for 3-(p-bromophenyl)-l-
methoxy-1-methylurea, a registered herbicide having the chemical
structure:
                                                    0CH3
Molecular formula:  C_H  BrN.O
Molecular weight:   259
Melting point:      95.5 to 96C
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

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                                 2                  Metobromuron EPA-3
                                                    (Tentative)
Reagents;
     1.  Metobromuron standard of known % purity
     2.  Aldrin standard of known HHON 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 pi
     4.  Usual laboratory glassware

Operating Conditions for TCP;
     Column temperature:     165C
     Injection temperature:  200C
     Detector temperature:   200C
     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 raetobromuron 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 rag 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 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 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.

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                                                         Metobrotnuron EPA-3
                                                         (Tentative)
         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 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:


     7 o (wt. HHDN)(% purity HHDN)(pk. ht. or area metobromuron)(100)
       " (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.

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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:
                     HC
                     HC
 H
Molecular formula:  C.H.N.CL
                     4422
Molecular weight:   112.1
Melting point:      296 to 298C; the technical product is at least
                    97% pure and has a m.p. of at least 292C.
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 dimethylformamide
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

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                                 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 ug 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 absprbances and using the standard and

sample concentrations, calculate the percent MH as follows:


 0 (abs. sample)(cone, std in ug/ml)(% purity std)
    (abs. std) (cone, sample in jig/ml)

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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:
                       o               V         y
                       P 0  C=C  C  N  CH-7
                                                          3
                                  CH3
Molecular formula:  C H ,NO P
Molecular weight:   223
Melting point:      54 to 55C (technical material 25 to 30C)
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-methyl-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, arid 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
           _i
     870 cm   (10.6 ji to 11.5 ji) .
          Determine the absorbance of standard and sample using the
     peak at 900 cm   (11.1 ja) and basepoint at 920 cm   (10.86 ji) .

     Calculation:
          From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent monocrotophos as
     follows:

      B (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.

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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:

                      P 0 C=C C N CH3
        CH3 CK             I           II
           J                    CH3       0
Molecular formula:  C-H..NOrP
                     7 14  5
Molecular weight:   223
Melting point:      54 to 55C (technical material 25  to  30C)
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

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                                 2                     Monocrotophos EPA-2
Other names:  Azodrin (Shell); Nuvacron (Ciba); Monocron; dimethyl-1-
              methyl-2-raethyl-carbamoyl-vinyl phosphate; cis-3-
              (dimethoxyphosphinyloxy)-N-methylcrotonamide;  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:     175C
     Injection temperature:  225C
     Detector temperature:   240C
     Carrier gas:            Nitrogen
     Carrier gas flow rate:  50 ml An in
     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 rag 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)

     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 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.

-------
                                                      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


      P _ (wt. 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:


         (wt. IS)(% purity IS)(pk. ht. or area monocrotophos)(100)
     8 "" (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.

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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:      174-175C
Physical state, color, and odor:  odorless, white crystalline solid
Solubility:  230 ppm in water at 25C; sparingly soluble in petroleum
             oils and in polar organic solvents; 5.2% in acetone at
             27C
Stability:   stable toward moisture and oxidation at RT but is
             decomposed at 185-200C; 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.

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                                 2                       Monuron 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 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 200C
     4.  Potentiometric titrimeter
     5.  Usual laboratory glassware

Procedure;
     Weigh a portion of sample equivalent to 0.4-0.5 gram monuron 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.

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                                                        Monuron EPA-1
     Pipette 50 ml of the 0.IN 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 175C  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:

     % 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. 24, 1475 (1952).

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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.-CULO
Molecular weight:   198.6
Melting point:      174-175C
Physical state, color, and odor:  odorless, white crystalline solid
Solubility:  230 ppm in water at 25C; sparingly soluble in petroleum
             oils and in polar organic solvents; 5.2% in acetone at
             27C
Stability:   stable toward moisture and oxidation at RT but is
             decomposed at 185-200C; 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

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                                 2                     Monuron EPA-2
Equipment;
     1.  Ultraviolet spectrophbtometer, 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 jug 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

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                            3                        Monuron EPA-2


methanol in each cell.  Scan both the standard and  sample  from
300 tun to 200 nm with methanol in the reference cell.
    Measure the absorbance of standard and  sample at 245 nm.

Calculation;
    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 jag/ml)

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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:      174-175C
Physical state, color, and odor:  odorless,white crystalline solid
Solubility:  230 ppm in water at 25C; sparingly soluble in petroleum
             oils and in polar organic solvents; 5.2% in acetone at
             27C
Stability:   stable toward moisture and oxidation at RT but is
             decomposed at 185-200C; 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

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                                 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 ji).
         Determine the absorbance of standard and sample using the
     peak at 1360 cm   (7.35 ;i) and baseline from 1380 cm   to 1325 cm~
     (7.25 u to 7.55 u).

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                                                Monuron EPA-3
Calculation;
    From the above absorbances and using the standard and
sample solution concentrations, calculate the percent monuron
as follows:

 a (abs. sample)(cone, std in mg/ml)(% purity std)
    (abs. std)(cone, sample in mg/ml)

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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

                       H     0    CH3
               y	N	CNCH2CH2CH2CH3
Molecular formula:  C,_H,,C10N00
                     i/ ID   /  /
Molecular weight:   275.18
Melting point:      102 to  103C
Physical state, color,  and  odor:  odorless, white crystalline solid
Solubility:  4.8 ppm in water  at  24C; 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

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                                                       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 /i to 8.5 /i).
         Determine the absorbance of the standard and sample using the
     peak at 1289 cm   (7.76 ji) and basepoint 1319 cm    (7.58/i).

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                                                         Neburon EPA-1
                                                         (Tentative)
     Calculation;

         From the above absorbances and using the standard and sample

     solution concentrations, calculate the percent neburon as

     follows:


      _ (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.

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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,
                     7 D 2 5
Molecular weight:   198.1
Melting point:      86C
Physical state, color, and odor:  yellowish, odorless, crystals
Solubility:  130 ppm in water at 15C; 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 85C
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

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                                 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 100C)
         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
     4.  Concentrated hydrochloric acid
     5.  Potassium iodide, 15% solution in water
     6.  Starch indicator solution
     7.  Sodium thiosulfate, 0.1N (or 0.3N) standard solution

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                                                    Nitrophenols EPA-1
Equipment;
     1.   300 ml  Erlenmeyer flask with rubber stopper  fitted with a
         Bunsen  valve  (described below)
                              B
                             -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
                              j
pressure to be relieved by allowing
gases to escape,  but is  sealed as out-
                              r
side pressure pushes in  since the sides
of the slit are pressed  together.
             LyJ
     2.  Water bath,  95100C
     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 100C  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,

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                                 A                     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 SnCl9 (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 milliequivalents of DNOC in 10 ml of sample aliquot.


       ..  .        ,  .      .    (meqs. of DNOC in 10 ml)(100)
     i dinitrocresol in sample = (grams sampie)(10/250)	

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                                                  Nitrophenols EPA-1
Chemical Reactions:
     Bichromate equivalent of stannous  chloride:
6SnCl
28HC1
            + 6KI -f 14HC1
                                     6SnCl  + 4KC1 + 4CrCl  + 14H-0
                            8KC1 + 2CrCl
     Sample reaction with  stannous chloride:
            N02                    HO


          f 6SnCI2 -I-  UHCI  	5
                                                 NH-HCI
                                                  6S>CI4 +- 4H20
                                             NH-HCI
     Oxidizing compounds with sodium thiosulfate:




     (oxidizing compounds) + KI  HC1 >  I
                       .A  2NaI + Na_S.O,
                       '          L 4 o

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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
                     / O f. _>
Molecular weight:   198.1
Melting point:      86C
Physical state, color,and odor:  yellowish, odorless, crystals
Solubility:  130 ppm in water at 15C; 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 85C
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

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                                 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 (-NO.) to amino  (-NH2) 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

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                                 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^O^.)
    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

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                                                      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 equal 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.

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                            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 bath
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 watchmaking 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

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                            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 ZnCL + 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.

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                                                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:

7 _ (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)
J _ _ *  "> _ *  ^ _
                            (grams of sample)*
v M-..   T-   11         j   / nitrogen in sample
% Nitrophenolic compound = ~c	:	j1-r	;-;	j
                           % nitrogen in nitrophenolic compound
* If extraction-cleanup procedure was used,, a dilution  factor  of
  100/200 must be added here.

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November 1975
Norborraide 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-
dicarboximide, a registered rodenticlde having the chemical structure:
                                   N
                                                         NH
Molecular formula:  C33H2sN3^3
Molecular weight:   511.6
Melting point:      180 to 190C (190 to 198 on crystals from methylene
                    chloride -f ether)
Physical state and color:  white to off-white crystalline powder
                           (mixture of isomers)
Solubility:  60 ppm in water at RT; at 30C 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)

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                                 2                     Norbormide EPA-1
Reagents:
     1.  Norbormide standard of known % purity
     2.  Chloroform, pesticide or spectro grade
     3.  Sodium sulfate, anhydrous, granular
     A.  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
     4.  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 jig/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 ug norbormide/ml)

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                            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:

y = (abs. 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) (2/100) (100)
  " (abs. std)(wt. sample)(1/100)(10/100)

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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)                     neral  (citral b)


                 CHO                      OHO
                  CH3
Molecular formula:  C,ftH,,0
                     lu ID


Molecular weight:   152.23





geranial  is a light oily liquid with a strong lemon odor; b.p._ , 92-93*C;
                                                              &  0

           20          20
          d;  0.8888; nT  1.48982; practically insoluble  in water; mlsclble



          with alcohol, ether, benzyl benzoate,  dlethyl phthalate, glycerol,



          propylene glycol, mineral oil, essential oils





neral     is a light oily liquid; lemon odor not as intense but sweeter



          than geranial; b.p.,, fi 91-92; dj 0.8869; nj  1.48690; solu-



          bilities same as geranial

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                                                    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 chromatpgraph 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 pi
     4.  Usual laboratory glassware

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                                 3             Oil of Lemongrass EPA-1
                                               (Tentative)
Operating Conditions for TCP;
     Column temperature:     150C
     Injection temperature:  250C
     Detector temperature:   250C
     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 yug/tnl)

     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 jig 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 40-50C just until the
     freons are removed.  This will minimize loss of any volatile con-
     stituents from the oil of lemongrass.

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                                               Oil of Lemongrass EPA-1
                                               (Tentative)
     Determination:
         Using a precision liquid syringe, alternately inject three
     20-30 jul 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:

     2 a (pic, 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 105C with a flame
ionization detector and nitrogen for carrier gas.

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  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
                                         .CH2-CH2-CH2-CH3
                 CH3  C  0  Sn -CH2-CH2-CH2-CH3
                                       \H2-CH2-CH2 CH3
       (b) bis  (tributyltin) compounds:

           example:  bis (tributyltin) oxide
CH3-CH2  CH2-CH2                   CH2-CH2-CH2-CH3
                             n 0 Sy,CH2 CH2 CH2-CH3
CH3-CH2 CH2-CH2                   CH2CH2 CH2-CH3

       In general, these compounds are practically insoluble in water
   but are miscible with organic solvents 1  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.

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                                 2                Organotin Compounds EPA-1
Principle of the Method;
     The orgariotin 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.IN standard solution - Prepare and
                       standardize as described under procedure.

Equipjoent.;
     1.  Kjeidahl flask and digestion set-up
     2.  Hot plate

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                                      3                 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.

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                            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.

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                            5                 Organotin Compounds EPA-1

Calculation;
    Calculate the normality of the potassium iodate solution as
follows:
    ...   (grams tin standard)
        (ml KIO,)(.05935)
    milliequivalent weight of tin = 0.05935
    Calculate the percent tin and organotin compound in the
sample as follows:
    X tin- C1H03)
             (grams sample)

    % Organotin compound * % tin x factor  (e.g., 2.511 for
                                     Bis(tributyltin)oxide)

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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:
                                    0
                             osv          yci
                                   o
Molecular formula:  C .HgCl.O.S
Molecular weight:   303.2
Melting point:      86.5C  (pure); about 80C (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

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                                 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
                                         9
         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)

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                                 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 /m to 13.5 jj).
         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 13.25ft).

     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.0 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.

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                                 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 1R determination.  Interfering substances
     may or may not be present as shown by a normal or distorted
     IR curve.

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December 1975                                           Parathlon 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:
     CH3CH2
     CH3	CH2	0
Molecular formula:  C QH ,NO_PS
Molecular weight:   291.3
Melting/boiling point:  m.p. 6.0C,  b.p.  157  to  162C  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 25C; 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 25C); isomerizes on  heating to the OS-dlethyl
             isomer

Other names: ACC 3422, Thlophos (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

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                                 2                        Parathion EPA-1
                                                          (Tentative)
Reagent 8:
     1.  Parathion standard of known % purity
     2.  Methanol, ACS

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.  235 nm has been found useful for parathion.
     2. . Suitable column such as:
         a.  DuPont ODS Permaphase, 1 meter x 2.1 mm ID
         b.  Perkin-Elraer 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:  50-55C
     Chart speed:         5 min/inch or equivalent
     Flow rate:           0.5 to 1.5 ml/rain (Perkin-Elraer 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.

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                                 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 jjg/ul)

     Preparation of Sample:
         Weigh an amount of sample equivalent to 0.3 gram parathion into
     a glassy-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:

      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,

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December1975
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	0
    CH3	CH20
Molecular formula:  C,_H,.NO-PS
                     10 14  5
Molecular weight:   291.3
Melting/boiling point:  m.p. 6.0C,  b.p.  157  to  162C  at 6 ram 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 25C;  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 25C); 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, Apharaite, Corothion,
             Ethyl Parathion, Etilon, Fosfono,  Orthophos,  Panthion, Para-
             mar, Paraphos, Parathene, Parawet, Phoskil, Rhodiatox,
             Soprathion, Strathion

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                                 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:  A1 x 2 mm I.D. glass column packed with 3% OV-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:     190C

     Injection temperature:  240C

     Detector temperature:   240C

     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.

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                                                         Parathlon 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 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 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.

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                                                        Parathion 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 parathion)
          (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 0 (wt.  alachlor) (% purity alachlor) (pk. ht. or area parathion) (100)
         (wt.  sample)(pk. ht. or area alachlor)(RF)
*
 The following columns also seem satisfactory:
     (1) A1 x 2 mm I.D. glass, packed with 5% SE-30 on 80/100 mesh
         Chrotnosorb W HP at 170C
     (2) 4' x 2 mm I.D. glass, packed with 5% OV-210 on 80/100 mesh
         Chromosorb W HP at 180C
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'a Beltsville Chemistry Lab.  Any comments, criticisms,
       suggestions, data, etc. concerning this method will be appreciated,

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 December 1975                                          Pebulate EPA-1
                                                        (Tentative)
                    Determination of Pebulate by

                  Gas-Liquid Chroraatography (TCD)


      Pebulate is the common name for S-propyl butylethylthiocarbamate,

 a registered herbicide having the .chemical structure:
                               0          ,0)2013

CH3CH2CH2 S C N<\

                                         \H2CH2CH2  CH3
 Molecular formula:  c10H2iNOS

 Molecular weight:   203.A

 Boiling point:      142C at 21 mm

 Physical state, color,  and odor:  clear yellow liquid with  an amine-

                                  like odor

 Solubility:  60 ppm in  water at 20C; 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 chroraatograph 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 jml
     4.  Usual laboratory glassware

Operating Conditions for TCD;
     Column temperature:     160C
     Injection temperature:  185C
     Detector temperature:   185C
     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 rag 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:
     y 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-pebulate)                     9.5 min.
            NjN'-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         /CH2CH3
                               11      /
CH3CH2CH2SCN<^

                                           CH2CH2CH2CH3
 Molecular formula:  C 1f.H_ NOS
 Molecular weight:   203.A
 Boiling point:      142C at 21 mm
 Physical state,  color, and odor:  clear  yellow liquid with an amine-like
                                  odor
 Solubility:   60  ppm in water at 20C;  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.  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 A 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 jil
     4.  Mechanical shaker
     5.  Centrifuge or filtration equipment
     6.  Usual laboratory glassware

Operating Conditions for FID:
     Column temperature:     150C
     Injection temperature:  225C
     Detector temperature:   250C
     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 A mg pebulate
     and A 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 A mg pebulate and
     A mg cycloate/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/A 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)(100)
         (wt. sample)(pk. ht. or area cycloate)(RF)
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         /CH2 CH3
CH3  CH2  CH2  S C  N<
                                           CH2 CH2  CH2  CH3
 Molecular formula:   C QH  NOS
 Molecular weight:    203.4
 Boiling point:       1A2C 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.  S-Ethyl dipropylthiocarbamate (EPTC) standard of known % purity

-------
                                 2                         Pebulate EPA-3
                                                           (Tentative)
     3.  Acetone, pesticide or spectro grade
     4.  Internal Standard solution - weigh 0.1 gram EPIC into a 50 ml
         volumetric flask; dissolve in  and make to volume with acetone.
         (cone 2 mg EPIC/ml)

Equipment:
     1.  Gas chromatograph with flame ibnization detector (FID)
     2.  Column:  4' x 2 mm 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:     130C
     Injection temperature:  180C
     Detector temperature:   180C
     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 by pipette 20 ml of the internal
     standard solution and shake to dissolve,  (final cone 3 ing pebulate
     and 2 mg EPTC/ml)

     Preparation of Sample:
         Weigh a portion of sample equivalent 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 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 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)(pk. ht,. or area pebulate)
          (wt. pebulate)(% purity pebulate)(pk. ht. or area EPIC)
         Determine the percent pebulate for each injection of the

     sample-internal standard solution as follows and calculate the

     average:


       _ (wt. EPTC)(%: purity EPTC)(pk. ht. or area pebulateHlOO)
         (wt. sample)(pk. ht. or area EPTC)(RF)
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
mol. wt. 170.20
 OH
white flaky crystals;  mild characteristic  odor; mp 55.5-57.5C:

bp 280-284C; practically insoluble in water,  soluble in alkali

hydroxide solutions and most organic solvents. Other names:

Dowcide 1, o-hydrodiphenyl, orthoxenol
]?-tert-butylphenol   C QH ^0   mol.  wt.  150.21
                                OH
                         CH3-C-CH3

-------
                                2          Phenols and Chlorophenbls EPA-1





white crystalline  solid; distinctive odor;  mp 98-100C; bp 237-239C;



practically insoluble  in cold water, soluble in alcohol, ether.



Other names:  Butylphen
p-tert-amylphenol    C-.H.-O   mol. wt. 164.24
                     11 16
                               OH
                       CH3-C-CH2-CH3

                              CH3
white crystals (irritating to skin); mp 94-95C;  bp  262.5C;



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 190-191C;



bp about 309-310C 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   C0HnC10    mol. wt. 156.61
	p-	o y
                                 OH
crystals with phenolic odor; mp 11.5.5C; bp 2A6C: volatile with steam;

one gram dissolves in 3 liters of water at 20C; 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-chlbro-3,5-dimethylphenol, 2-chloro-

5-hydroxy-l,3-diraethylbenzene
o-benzyl-p-chlorophenol   C.-H.-CIO    mol. wt. 218.69
		j^j _^
                        OH               	

                        Cl

white to  light tan or pink flakes; slight phenolic odor; mp 48.5C;

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

-------
4-chloro-2-phenylphenol
         OH
Phenols and Chlorophenols EPA-1

6-chloro-2-phehylphenol
           OH
            mol. wt. 204.65
clear colorless to straw-colored viscous liquid
  with faint characteristic odor
boiling range .5-95%   146-158.7C  (5mm)
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)

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March 1976                                Phenols and Chlorophenols F.PA-2

         Determination of o-Phenylphenol in Disinfectant
             Formulations by Ultraviolet Spectroscopy

     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 % purity
     2.  Sodium hydroxide, IN aqueous solution
     3.  Ethanol, ACS
     4.  Hexane, purified.  Extract 250 ml n-hexane with two 20 ml
         portions 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;
     Preparation of Standard;
         Weigh 0.04 gram o-phenylphenol standard into a 100 ml volumetric
     flask, add 5 ml IN sodium hydroxide solution, dissolve and dilute

-------
                            2     Phenols and Chlorophenols FPA-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 ug o-phenylphenol/ml)

Preparation of Sample:
    Weigh a portion of sample equivalent to 0.008 gram o-nhenyl-
phenol into a 125 ml Erlenmeyer flask, add 1 drop of 1\? sodium
hydroxide, and evaporate to dryness on a rotary evaporator.
Dissolve the residue in about 40 ml water anrl 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 separatory 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 nil 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 50 ml volumetric flask,
add 3 ml IN sodium hydroxide solution and 25 ml ethanol, dilute to
volume Xi/ith water, and mix thoroughly.  (final cone 16 ug o-phenyl-
phenol/ml)

-------
                                       Phenols and Chlorophenols EPA-2
]N_ Determinat ion:
    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 312 nm with 0.1N
sodium hydroxide solution in each cell.  Scan both the standard
and sample from 360 nm to 260 nm with 0.1N 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:

7 = (abs. sample)(cone. std in |ig/ml)(% purity std)
    (abs. std) (cone, sample in

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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
solutions; 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

-------
                                 2        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
             V
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  =  	,	r^-r	
                              (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 Broraination 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-phenylpheriol 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
         brornate and 15 grams potassium bromide in water and make to one
         liter.  This solution heed not be standardized if a blank using
         the same quantity as the sample is run each time.
     4.  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 ral 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. IN .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.  Shake 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 endpoint.
    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 tit-ration. -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 NaO. (N NaO.  (0.04255) (100)
                      (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.S_0.)(N Na.S00_) (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

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                                       Phenols and Chlorophenols EPA-4
Reactions:
1.  Release of  bromine from bromate-bromide  solution:
KBrO  + 6KBr +  6HC1
                                     + KBr  + 6KC1 +
2.  Bromination of  the o-phenylphenol:
        ONa
                             ONa
                                                            2HBr
3.  Release of  iodine from excess bromine:
         (excess)Br  + 2KI	>I  + 2KBr
A.  Titration of  iodine with sodium thiosulfate;
         I + 2Na0S00.	>2NaI + Na.S.O,
          L     L  i  J              2 4 b

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March 1976                                    Phenols and Chlorophenols EPArS?.
                                                                  (Tentative
                Determination of Pentachlorophenol
              by High Pressure Liquid Chrpmatography

     For definition, structure, and technical data on pentachlprophenol,
see Phenols and Chlorophenols EPA-1.

Reagents;
     1.  Pentachlorophenol standard of known % purity
     2.  Benzyl benzoate standard of known % purity
     3.  Ethanol, ACS
     A.  Internal standard solution - weigh 5 grams benzyl benzoate
         into a 50 ml volumetric flask; dissolve in and make to
         volume with ethanol.  (cone 100 rag/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:  60C
     Chart speed:         12 inches/minute
     Flow rate:           1.0 ml/minute

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                                             Phenols and Chlorophenols EPA-5
                                                                 (Tentative)
     Pressure:     800-1000 psi
     Detector:     UV at 254 run
     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.

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                                             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:

      = (wt. IS)(% purity IS)(pk. ht. or area PCP)(100)
     "   (wt. sample)(pk. ht. or area IS)(RF)
Method submitted by Elmer H. Hayes, EPA, Beltsville, Md.

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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:  250C
     Detector temperature:   250C

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                                              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

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                                            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/ml for o-benzyl-p-chlorophenol.)

     Determination;
         A column temperature of 180C is sufficient for o-phenylphenol
     and p-tert-amylphenol, eluting in that order.  However, a 220C
     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 150C to 250C.

     Calculation:
         From the average peak height or peak area calculate the percent
     phenol or chlorophenol compound 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, OPP, TSD, Beltsville, Md.

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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 jil or 50 /il
     5.  Usual laboratory glassware

Procedure using Thermal Conductivity Detector;
     Operating Conditions for TCP;
         Column temperature:     210C
         Injection temperature:  240C

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                                        Phenols and Chlorophenols EPA-7
                                                            (Tentative)
    Detector temperature:  270C
    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)
        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)

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                                                 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:

     "/  (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:     145C
         Injection temperature:  225C
                                                                i
         Detector temperature:   220C
         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.

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                                           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 of 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 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 4-chloro-3,5-xylenol as follows:

     
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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

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                                            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 chroroatograph 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:     165C

     Injection temperature:  215C

     Detector temperature:   230C

     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.

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                                           Phenols and Chlbrbphenols 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 mg/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 Erlenraeyer 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

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                                 4         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 terminat 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.

                                            1 ml internal standard
2 ml phenol compound (20 mg)                	(mg as  listed)	
p-tert-butylphenol                          n-tetradecane  20 mg
p-tert-amylphenol                           n-tetradecane  20 mg
o-phenylphenol                              lindane  80 mg
4-chloro-2-cyclopentylphenol                n-hexane  10 mg
o-benzyl-p-chlorophenol                     benzyl benzoate  12.5 mg
4 or 6-chloro-2-phenylphenol                benzyl benzoate  12.5 mg
2,2'-methylenebis(3,4,6-trichlorophenol)    di-2-ethylhexylphthalate(A)  10 mg
                                               10 mg
2,2'-raethylenebis(4-chlorophenol)           di-2-ethylhexylphthalate(B)  40 mg
                                               40 mg

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                                    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.
 T   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,
allowing 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 phenol)(100)
    (wt. sample)(peak height of internal standard)(RF)

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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:      185C, sublimes 130C (1 mm); b.p. 371C
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

-------
                                                      Phenothiazlne 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
     4.  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 rag/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)

-------
                                                  Phenothiazine EPA-1
                                                  (Tentative)
IR Determination:
    With carbon dlsulfide 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 p. to 8.0 ji).  For a qualitative
comparison, run a full scan.
    Determine the absorbance of standard and sample using the
                             mm *
minimum absorbance at 1333 cm   (7.5 u) and the maximum absorbance
at 1299 cm"1 (7.7 i).
Calculation;
    From the above absorbances and using the standard and sample
concentrations, calculate the percent phenothiazine as follows:

  (abs. sample)(cone, std in mg/ml)(% purity std)
    (abs. std)(cone, sample in mg/ml)

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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:
     Q. n -3 L. M o    U N.    o
        3      2     \ II      _    	
                             ~~~3    v,r1o  ~ ^    V-H'^  "v-rl-
         7CH20'
Molecular formula:  C?H _0 PS
Molecular weight:   260.4
Melting and boiling point:  mp  -42.9C (pure material)
                            bp  118-120C at 0.8 mm Hg
Physical state and color:   clear mobile liquid
Solubility:  50 ppm in water at RT; raiscible 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

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                                 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)

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                                 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 40C 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 cm"1 (13.7 ^i to 16.9 ji) .
         Determine the absorbance of standard and sample using the
     peak at 654 cm   (15.3 ji) and baseline from 709 cm"  to 599 cm"
     (14.1 ji to 16.7 ^i).

     Calculation:
         From the above absorbances and using the standard and sample
     solution concentrations, calculate the percent of phorate as
     follows:
     a, _ (abs. 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.

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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) :
                                                Ov   0
Phosphate:  (dicrotophos)                              yP _ 0 _ R
                                           CH3 0'

                                    CH3-CH2-0    S
Phosphorothioate:   (parathion)                         /P  0  R
                                         -CH CT
                                    CH3_CH2-0    S
Phosphorodithioate:   (phorate)                        ^p  S  R
                                    CH3-CH2 CT
     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.

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                                 2                    Phosphorus Compounds EPA

     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 44.1C, bp 280C (volatile, sublimes in
                             vacuo at ordinary temperature when exposed
                             to light)
     Physical state and  color:    white or yellow,  soft waxy solid 

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                                 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 raolybdic 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 65C deposits no yellow pre-
         cipitate of ammonium phosphomolybdate.  An alternative procedure
         is to heat to 65C for 1-2 hours and allow 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.

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                                 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 phosphomolybdate
         precipitate)
     9.  Platinum Gooch crucible
    10.  Asbestos, acid and alkali washed (preferably pre-ignited at
         900-1000C before washing)
    11.  Muffle furnace
    12.  Usual laboratory glassware

Procedure:
	                                                 V
     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.

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                            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 pov/ders.  For the analysis of organophosphates
in granules, dusts, or wettable powders,it is recommended that the
sample be extracted with chloroform.  This will simplify 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 petroleum 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
-------
                            b                Phosphorus Compounds EPA-1

    For Organophosphorus 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.

Precipitation as Ammonium Molybdate:
    Add ammonium hydroxide to slight alkalinity and then make dis-
tinctly acid with nitric acid.  Heat to 65C and add 70 ml of ammonium
molybdate solution.  Stir and digest at 65C 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.

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                            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
allow 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 900-1000C.  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 900-1000C
until the residue is white.  Cool and weigh as magnesium pyrophosphate.

Calculation:
    From the weight of magnesium pyrophosphate, Mg-P-0_, calculate
the percent phosphorus in the sample as follows:

               (grams Mg^ty (0.2783) (100)
% phosphorus = 	;	ir	
                      (grams sample)

0.2783  =  factor MgP0  to phosphorus

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                                               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:
Organophosphate  - - - > H^POA  +  Oxidation product
                       21HN03
(NH,)3P04.12Mo03 + 2ANH.OH 	^  (NH^) PO^ + 12(NH^)2MoO, +
(NH.),PO. + MgCl. + NH.OH 	>  MgNH.PO. + 2NH.C1 + NH.
   4342     4          ^44      4       4

OH

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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:
                            NH2
                 Ck    /^\    JC\
Molecular formula:  C,H_C1.N000
                     D J  J i L
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 25C; 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 215C; subject to photo-
                decomposition

Other names:    Tordon (Dow Chem. Co.), Borolin
Reagents;
     1.  Picloram standard of known % purity
     2.  Methanol, pesticide or spectro grade

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                                 2                           Picloram EPA-1
                                                               (Tentative)
Equipment;
     1.  High pressure liquid chromatograph
         t
     2.  High pressure liquid syringe or sample injection loop
     3.  Liquid chroraatographic 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:  65C
     Column pressure:     900 PSI
     Flow rate:           8 ml/min
     Chart speed:         10 rain/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).

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                                 3                          Picloram EPA-1
                                                             (Tientative)
         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:


      a (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.

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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:
                                         0
                                  CH  C -
Molecular formula:  C ,H ,0
Molecular weight:   230.3
Melting point:      108.5 to 110.5C
Physical state and color:  yellow crystalline solid
Solubility:  18 ppra in water at 25C; 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.

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                                 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 ran.  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 110-111C
and PMP melts at 67-68C.
     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,P20_.10H20 in 500 ml water.
     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

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                                 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 jig pindone/ml)

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                                                    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 nra and
calculate the percent pindone as follows:

., _ (abs. sample) (cone, std in ug/ml)(% purity std)
    (abs. std)(cone, sample in ug/ml)

or using dilution factors, as follows:

z   (abs. sample)(wt. std)(purity std)(1/100)(10/100)(5/100)(100)
    (abs. std)(1/200)(2/10)

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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:
                                         0
                                 CH	C-
Molecular formula:  ciAHiA3
Molecular weight:   230.3
Melting point:      108.5 to 110.5C
Physical state and color:  yellow crystalline solid
Solubility:  18 ppm in water at 25C; 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.

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                                 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, recrystallizatlon from pentane,
and determination of the melting point.  Pindone melts at 110-111C
and PMP melts at 67-68C.
     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

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                                 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
                                                      4
     cell.  Scan both standard and sample from 350 nm to 200 nm with
     the pyrophosphate solution in the reference cell.

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                                                   Plndone  EPA-2
Calculation;

    Measure the absorbance of standard and sample at  283  ntn and

calculate the percent pindone as follows:


7 0 (abs. sample)(cone, std inug/ml)(% purity  std)
    (abs. std) (cone, sample in fig/ml)


or using dilution factors, as follows:


y   (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

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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,.0_
                     14 14 3
Molecular weight:   230.3
Melting point:      108.5 to 110.5C
Physical state and color:  yellow crystalline solid
Solubility:  18 ppm in water at 25C; 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 (Kllgore 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,

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                                 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 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 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 110-111C and PMP melts at 67-68C.



     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.P00_.10H-0 in one liter of water.
           427    2




Equipment;



     1.  Ultraviolet spectrophotometer, double beam ratio recording


         with matched 1 cm cells



     2.  Usual laboratory glassware

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                                 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 jig 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 hm 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.

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                                                    Pindone EPA-3
Calculation:
    Measure Che absorbance of standard and sample at 324 run
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 agentEDTAit remains color-
less or becomes only very faintly pink.

-------
                                                     Plhddne EPA-3
    For Benzoate - wash the residue with hot water to remove the
benzole 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 benzole acid.  An alterna-
tive procedure is to evaporate the acidified filtrate and
determine the melting point.  Benzole acid melts at 122C.

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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:  C1(jH_nO,
Molecular weight:   338.5
Boiling point:      180C 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

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                                 2                  Piperonyl Butoxide EPA-1
Other names:  Butacide (FMC), NIA 5273 (Niagara), FMG 5273, cr-[2-(2-n-
              butoxyethoxy)-ethoxy]-4,5-methylenedioxy-2-propyltoluene

Reagents:
     1.  Tannir. acid
     2.  Acetic acid, glacial
     3.  Phosphoric acid, 85%
     A.  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.

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January 1976
                      Piperonyl Butoxide  EPA-2
              Determination of Piperonyl Butoxide
                  by Gas-Liquid Chromatography
                   (FID - Internal Standard)

     Piperonyl butoxlde, 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:
      .o
      '0
CH2-CH2-CH3

 CH2-0 -CH2-CH2-0- CH2-CH2-0- <
Molecular formula:  C  H  0
Molecular weight:   338.5
Boiling point:      180C 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

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                                 2               Piperonyl Butoxide EPA-2

Other names:  Butacide (FMC), NIA 5273 (Niagara), FMC 5273, a-[2-(2-n-
              butoxyethoxy)-ethoxy]-4,5-methylenedioxy-2-propyltoluene

Reagents;
     1.  Piperonyl but'oxid'e of known % purity
     2.  Dioctyl phthalate
     3.  Acetone, pesticide or spectro grade
     4.  Internal Standard solution - weigh 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 chromatograph with flame idnization detector (FID)
     2.  Column:  61 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:     230C
     Injection temperature:  260C
     Detector temperature:   270C
     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)

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                                 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;
     Preparation 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.

-------
                                                   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 phthalate

__   (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:

      _ (wt. I.S.)(% purity I.S.)(pk. ht. or area piperonyl butoxide)(100)
         (wt. sample)(pk. ht. or area I.S.)(RF)
Method submitted by Division of Regulatory Services, Kentucky Agricultural
Experiment Station, University of Kentucky, Lexington, Kentucky A0506.

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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:
                                         0
                                      C	CH2CH
Molecular formula:  C, ,H,. 0_
                     14 14 3
Molecular weight:   230.3
Melting point:      67 to 68C
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.

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                                 2                         PMP   EPA-1

     The method does not distinguish between pindbne (2-pivalyl-l,3-

indandione) and PMP (2-isoyaleryl-l,3-indandione), both of which have

UV absorption maxima at 283, 312, and 32A run.  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 110-111C
                                      i
and PMP melts at 67-68C.

     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

         NavP26_.10H20 in 500 ml water.

     3.  Ethyl ether, ACS

     A.  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.  Spxhlet extraction apparatus

     3.  Mechanical shaker

     A.  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 ug 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)

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                                                     PMP   EPA-1
UV Determination;
    With the UV spectrophotoineter 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 PMP as follows;

 = (abs. sample)(cone,  std in ug/ml)(% purity std)
    (abs. std) (cone, sample in jag/ml)

or using dilution factors, as follows:

v m (abs. sample)(wt. std)(purity std)(1/100)(10/100)(5/100)(100)
    (abs. std)(1/200)(2/10)

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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:
                                         0
                                      CCH2CH
Molecular formula:  ciAH-iAO->
Molecular weight:   230.3
Melting point:      67 to 68C
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 Chetn. Co.)

     This method may be used for analyzing both bait materials and
concentrates containing about 0.025% and 0.5% active ingredient.

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                                 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 110-111C
and PMP melts at 67-68C.
     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
         Na4P20?.10H20 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.
     A.  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 pg 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.

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                                                   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:


z m (abs. sample)(wt. std)(purity std)(1/100)(10/100)(5/100)(100)
    (abs. std)(wt. sample)(1/100)(20/50)(10/10)

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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:
                                 GH  CCH2CH
Molecular formula:  C..H..0.
                     14 14 3
Molecular weight:   230.3
Melting point:      67 to 68C
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 ethylenediaraine 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 fig/ml in 1% pyrophosphate
has an approximate absorbance of 0.394 at 324 nm; a solution of PMP
containing 7.5 iig/ral 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 110-111C and PMP melts at 67-68C.
     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_07.10H_0 in one liter of water.

Equipment;
     1.  Ultraviolet spectrophotometer, double beam ratio recording
         with matched 1 cm cells
     2.  Usual laboratory glassware

-------
                                 3                       PMP   EPA-3

Procedure;
     Preparation of Standard;
         Weigh 0.075 gram PMP standard tnto 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 pg 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 jig
     PMP/ml)

     Preparation of Sample:
         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 jig 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 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 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.

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                            4                    PMP   EPA-3

Calculation;
    Measure the absorbance of standard and sample at 324 nm
or 283 no,  and calculate the percent PMP as follows:

% PMP at 324 nm:

    '/ . (abs. sample)(wt. std) (purity std)(1/100)(5/50)(5/50)
      " (abs. std)(wt. sample)(1/100)(25/100)

% PMP at 283 nm:

    7 = (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 agentEDTAit remains colorless
or becomes only very faintly pink.

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                                                   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 salmoncolor 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 122C.

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November 1975
Prometone EPA-1
(Tentative)
                    Determination of Prometone
                   by GasLiquid 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 :
                                 0-CH3
Molecular formula:  C QH  N 0
Molecular weight:   225.3
Melting point:      91 to 92C; the technical product is at least 97%
                    pure and has a ra.p. of 88-90C
Physical state and color:  white crystalline solid
Solubility:  750 ppm in water at 20C; 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, GA1A35 (Ciba-Geigy); prometon (ISO),
             Gesafram, Outrack

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                                 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 pi
     4.  Usual laboratory glassware

Operating Conditions for TCD;
     Column temperature:     175C
     Injection temperature:  225C
     Detector temperature:   250C
     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.

-------
                                                      Proraetone EPA-1
                                                      (Tentative)
Procedure:
     Preparation of Standard;
         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 rag prometone and 10 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)

     Determination;
         Inject 2-4 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 proraetone, then heptachlor.
         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 prometone and heptachlor
     from both the standard-internal 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)(% purity heptachlor)(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:


     v = (wt.  heptachlor)(% purity heptachlor)(pk. ht. or area prometone)(100)
         (wt.  sample)(pk. ht. -or area heptachlor)(RF)
This method was developed by Stellos Gerazounis, EPA, Region II, New

York, N. Y., and was collaborated (with slight modification) by Elmer

Hayes, EPA, Beltsville Chemistry Laboratory, Beltsville, Md.

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January 1976
Prometone EPA-2
(Tentative)
                  Determination of Prometone by
                    Gas-Liquid Chroinatography
                    (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.QH.-N-O
Molecular weight:   225.3
Melting point:      91 to 92C; the technical product is at least 97%
                                                  r
                    pure and has a m.p. of 88-90C
Physical state and color:  white crystalline solid
Solubility:  750 ppm in water at 20C; 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:     180C
     Injection temperature:  230C
     Detector temperature:   230C
     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 intermittently for one hour,  (final cone 2 mg
     prometone and 5 mg alachlor/ml)

     Determination:
         Inject 1-2 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 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)(100)
         (wt. sample)(pk. ht. or area alachlor)(RF)
Note:  For an alternative procedure to the above method, the following
       changes can be made:
            solvent:  chloroform
            sample concentration:  1.6 mg/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.

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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
                                              H           CH2
                                               CH2-CH2
                                               0

                       HC=CCH20S=0
Molecular formula:   C1QH?,0,S
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 spectro 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 Standard;
         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)
         For liquid formulations and emulsifiable 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 60C 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.G.'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 ji to 3.2 ji) .
    Determine the absorbance of standard and sample using the peak
at 3300 cm"1 (3.03 u) and a baseline from 3356 cm"1 to 3247 cm
(2.98 ji to 3.08 ji).

Calculation:
    From the above absorbances, calculate the percent propargite
as follows:

 = (abs. sample)(cone, std in mg/ml)(% purity std)
    (abs. std)(cone, sample in mg/ml)

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November 1975                                           Propargite EPA-2
                                                        (Tentative)

                   Determination of Propargite
                   by Gas-Liquid Chromatography
                    (TCD - Internal Standard)

     Propargite is a common name for 2-(p-tert-butylphenoxy)cyclohexyl-
2-pro'pynyl sulfite, a registered acaricide.having the chemical structure:
                           = CCH20 S=0
Molecular formula:  C  H  0 S
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, D01A (Uniroyal); Comite

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                                 2                       Prbpargite 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',1 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 jil
     4.  Usual laboratory glassware

Operating Conditions for TCP;
     Column temperature:     220C
     Injection temperature:  250C
     Detector temperature:   250C
     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 jil 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:

        - (wt. HEOD)(% 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:

     % __ (wt. HEOD)(% purity HEOD)(pk. ht. or area propargite) (100)
         (wt. sample)(pk. ht. or area HEOD)(RF)
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,rtHQClN,0
                     1U o   j
Molecular weight:   221.6
Melting point:      207C with decomposition
Physical state, color, and odor:  yellowish-tan to brown powder,
                    odorless when pure
Solubility:  400 ppm in water at 20C, 2.8% in acetone, 3.4% in
             methanol, 0.07% in benzene and in ether, 0.21% in
             chloroform, 0.6% in ethyl acetate
                                4
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

-------
                                                        Pyrazbn 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 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)

     Determination;
         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 ji).
         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 ji to 12.55 ji).

-------
                                                       Pyrazon EPA-1
                                                       (Tentative)
     Calculation:
         From the above absorbances and using the standard and

     sample solution concentrations, calculate the percent pyrazon

     as follows:


      a (aba, 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.

-------
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  jasraolin 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
     The chemical structure of these compounds is as follows:
        \
   R
            H
C=CH3C
     CH3-C
            CH3
 V
I    H   0
 \l    II
   >CC0
"A
C       C-
             HC
                 H
                                                          R'
       C=0

pyrethrin I (C_ H.ftO_)
pyrethrin II (C22H285)
cinerin I (C20H280,)
cinerin II (Q21H2gO )
jasmolin I (C21H-JO1^
jasmolin II (C,9H.nO,.)
R
-CH
3
-CO-O-CH
-CH3
-CO-0-GH3
-CH3
-CO-O-CH.
                                                          R1
                                                   -CH -CH=CH-CH=CH2
                                                   -CH2-CH=CH-CH=CH2
                                                   -CH2-CH=CH-CH3
                                                   -CH2-CH=CH-CH3
                                                   -CH2-CH=CH-CH2-CH3
                                                   -CH2-CH=CH-CH2-CH3

-------
                                 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 dicarboxylic




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.

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                                 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.

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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
     A.  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:  5' 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.  Mechanical shaker
     6.  Centrifuge or filtration equipment
     7.  Usual laboratory glassware

Operating Conditions for FID;
     Column temperature:     190C
     Injection temperature:  205C
     Detector temperature:   205C
     Carrier gas:            Nitrogen
     Carrier gas flow 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
     jig/jil for pyrethrin I and l.l.pg/fil 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 jig, for PBO
0.6 to 5.6 jig, 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 rag
pyrethrin I for a final volume of 25 ml - 0.4 ug/pl) 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 jal of
standard and sample are alternately injected for pyrethrin I and
PBO.  Smaller amounts or an additional dilution is needed to keep
the NOBD within the linear range  (0.3-1.7 jag 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.

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                                                     Pvrethrins EPA-2
     Other pyrethfum 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.

     7         r   (peak ht. sample)(cone.  std)(pi ,std injected)(100)
          "        (peak ht. std)(conc. 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.

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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 monocarboxylic 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
     A.  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:
     li  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 0-5C 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)              c       , _.,      ,  ,
	*	*::	  , , ,.	   =   grams of non-volatile needed
         (% claim on label)                

 .       ,  . .,      (wt. can & contents after heating)-(wt. empty can)
 h non-volatile  =  	;	.. ,,	\;	"	v'*	
                              (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 ml 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.
    .     .  ,    (ml 0.02N NaOH)(N 0.02N NaOH)(.3284)(100)
    A pyrethrin = (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 100C for 10 minutes.  Blow
gently into the flask several times to remove vapors.
    Add 30ml 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 milliequivalerit weight of pyrethrin II is
                  0.1862.

                TT _ (ml 0.02N NaOH)(N 0.02N NaOH) (.1862) (100)
              n ii - (grams sampie)(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
                       RNCH3
                                                    Cl



                                  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
OH3
CH3-C-CH2-C
   3   I        '
                                                   I
                        0-CH2 CH2-0-CH2CH2-N-CH
       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.
                                                                                      Q

-------
                                             Quaternary Ammonium Compounds  EPA-:?;)
     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.
CH3(CH2)|(
                                                        C|
     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.

-------
                                    FACTORS FOR VARIOUS QUATERNARY COMPOUNDS
                                                                         M.W.   %HALOGEN(FACTOR)   % N  (FACTOR)
ALKENYL DIMETHYL ETHYL AMMONIUM BROMIDE
    %C18  %C16
     100
      90    10            (Onyxide)
      15    85            (ST-50)
      80    20            (LQ-750)

ALKENYL DIMETHYL ETHYL AMMONIUM CHLORIDE
     100%-C18
      90%-C18, 10%-C16

ALKENYL 1-HYDROXYETHYL-l-ETHYL IMIDAZOLINIUM BROMIDE
     100%-C12

ALKENYL TRIMETHYL AMMONIUM CHLORIDE
     100%-C18             (Aliquat 11)

ALKYL 1-BENZYL-l-HYDROXYETHYL IMIDAZOLINIUM CHLORIDE
     100%-C13

ALKYLBENZYL TRIMETHYL AMMONIUM CHLORIDE
    %C9   %C10  %C11  %C12  %C13  %C14  %C15
             4
             4
27
23
100
 56
 56
9
9
3
3
1
1
ALKYL DIMETHYL BENZYL AMMONIUM CHLORIDE
    %C12  %C14  %C16  %C18
                       100   (Onyx 4002)
     14     58    28
                 100         (Onyx T)
     10     60    30
           100               (Hyamine 1450, BTC 927)
     65     30     5
     50     30    17     3   (BTC 50, BQL 50, LC 5373)
C22HA6NBr
C22HA6NC1
C19H3?ON2Br
C21H44NC1
C25H430N2C1
404.53
401.7
380.7
398.9
360.07
357.3
389.42
346.04
423.09
19.75 %Br(5.062)
19.89 %Br(5.027)
20.99 %Br(4.764)
20.03 %Br(4.992)
9.846%C1(10.16)
9.923%C1(10.08)
20.52 %Br(4.873)
10.25 %C1(9.761)
8.380%C1(11.93)
3.463
3.487
3.679
3.511
3.890
3.921
3.597
4.048
3.311
(28.88)
(28.68)
(27.18)
(28.48)
(25.71)
(25.51)
(27.802)'
(24.71)
(30.21) '
                                                        C22H40NC1
                                      C27H50NC1
                                      C23H42NC1
354.02
351.6
350.5


424.16
372.0
396.10
382.1
368.05
351.2
360.5


10.01 ZCK9.986)
10.08 %C1(9.918)
10.11 %C1(9.887)


8.358%C1(11.96)
9.531%C1(10.49)
8.950%C1(11.17)
9.279%C1(10.78)
9.633%C1(10.38)
10.09 %C1(9.907)
9.835%C1(10.17)


3.957
3.983
3.996


3.302
3.765
3.536
3.666
3.806
3.988
3.886


(25.27)
(25.10)
(25.02)


(30.28)
(26.55)
(28.28)
(27.28)
(26.28)
(25.07)
(25.74)


0
IT
m
3
n
|
o
3
H-
3
C5
0
9
o
c
3
O.
CO
                                                                                                                    M

                                                                                                                    I

-------
ALKYL DIMETHYL  BENZYL AMMONIUM CHLORIDE (CONT.)
    7.C12  7.C14  7.C16   7.C18
                                                               M.W.    7.HALOGEN (FACTOR)
                                                                                                        N  (FACTOR)
    LOO
     61
     40
      5
      5
23    11     5     (LC. 6215)
50    10       (Hyamine  3500,  BQM 50,  MC 5410)
90     5          (Dibactol)
60    30     5     (BTC 824, MC 6355)
ALKYL DIMETHYLBENZYL DIMETHYL AMMONIUM CHLORIDE
    1007.-C12
     507.-C12,  307.-C14,  177.-C16, 37.-C18  (BTC 927)

ALKYL DIMETHYL 3,4-DICHLOROBENZYL AMMONIUM CHLORIDE
    7.C12  7.C14 %C 16 7.C18
     100
      50   30     17      3     (ADC-60, BQL-50)
      23   55     20      2
       5   60     30      5     (Guardsan 50-50)

ALKYL DIMETHYL ETHYL AMMONIUM BROMIDE
    100Z-C16
     501-C12.307.-C14,  177.-C16, 37.-C18

ALKYL DIMETHYL ETHYLBENZYL AMMONIUM BROMIDE
    100Z-C16
     50I-C12,  3W-C14,  177.-C16, 37.-C18

ALKYL DIMETHYL ETHYLBENZYL AMMONIUM CHLORIDE
    1007.-C16
     507.-C12,  307.-C14,  177.-C16, 37.-C18  (BTC 471)

ALKYL DIMETHYL FURFURYL AMMONIUM CHLORIDE
    1007.-C18
     507.-C14,  307o-Cl6,  207.-C18
      5%-C14,  307.-C16,  657.-C18
C H NCI
. JL J\J
')


C23H42NC1

C21H36C12NC1



G20H44NBr

C27H50NBr


C27H50C1N

C^cHi nNOCl









340
356
359
368
377
368
388
408
429
437
446

378
342
468
433

424
388
414
377
402







.00
.8
.6
.0
.9
.05
.5
.88
.4
.2
.8

.49
.9
.61
.0

.16
,5
.12
.6
.9







10
9.
9.
9.
9.
9.
9.
8.
8.
8.
7.

21
23
17
18

8.
9.
8.
9.
8.







.43
936
858
633
382
633
125
671
257
109
936

.11
.31
.05
.46

358
125
561
388
800







7.C1(9.
7.C1(10
7.C1(10
7.C1(10
7.C1(10
7.C1(10
7.C1(10
7.C1(11
7.C1(12
*C1(12
7.C1(12

7.Br(4.
%Br(4.
*Br(5.
7.Br(5.

IC1(11
%C1(10
%C1(11
7.C1(10
7.C1(11







590)
.06)
.14)
.38)
.66)
.38)
.96)
.53)*
.11)*
.33)*
.60)*

736)
291)
864)
418)

.96)
.96)
.68)
.65)
.36)







4
3
3
3
3
3
3
3
3
3
3

3
4
2
3

3
3
3
3
3







.120
.925
.895
.806
.707
.806
.605
.426
.262
.204
.135

.701
.085
.989
.235

.302
.605
.382
.709
.477







(24
(25
(25
(26
(26
(26
(27
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(30
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(27
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(27
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.27)
.47)
.67)
.28)
.98)
.28)
.74)
.19)
.65)
.21)
.90)

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.48)
.46)
.91)

.28)
.74)
.57)
.96)
.76)

















in



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c
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a
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7"

-------
ALKYL DIMETHYL NAPHTHYLMETHYL AMMONIUM CHLORIDE
    100I-C12
     98Z-C12, 2Z-C14
    100Z-C12, monohydrate
     98Z-C12, 2Z-C14, monohydrate

ALKYLDODECYLBENZYL TRIMETHYL AMMONIUM CHLORIDE
    100Z-C12
     95Z-C12, 5Z-C18         (DBC-50)

N-ALKYL N-ETHYL MORPHOLINIUM ETHYL SULFATE
    100Z-C12
     92Z-C12, 8Z-C16
     92Z-C8, 8Z-C16(Alkyl from soy beans)

ALKYL ISOQUINOLINIUM BROMIDE
    100Z-C12
     50Z-C12, 30Z-C14, 17Z-C16, 3Z-C18
     61Z-C12, 23Z-C14, 11Z-C16, 5Z-C18  (LIB 75)

2-ALKYL 1-METHYL 1-HYDROXYLETHYL IMIDAZOLINIUM CHLORIDE
    100Z-C13

ALKYL METHYL ISOQUINOLINIUM CHLORIDE
    100Z-C12
     25Z-C12, 55Z-C14. 17Z-C16, 3Z-C18  (Araroonyx 781)

ALKYLNAPHTHYLMETHYL PYRIDINIUM CHLORIDE
    100Z-C12

ALKYL TOLYLMETHYL DIMETHYL AMMONIUM CHLORIDE
    100Z-C12

ALKYLTOLYLMETHYL TRIMETHYL AMMONIUM CHLORIDE
    100Z-C12

ALKYL TRIMETHYL AMMONIUM BROMIDE
    100Z-C16
                                                                        M.W.    '/.HALOGEN (FACTOK)   '/. N  (FAi.'lQH)
C2
-------
 ALKYL TRIMETHYL AMMONIUM CHLORIDE
     1007.-C16
       5VC16, 95Z-C18

 BENZYL DODECYLCARBAMYLMETHYL DIMETHYL AMMONIUM CHLORIDE
                             (Urolocide)

 CETYL PYRIDINIUM BROMIDE
           monohydrate

 CETYL PYRIDINIUM CHLORIDE
           monohydrate

 2-CHLOROETHYL TRIMETHYL AMMONIUM CHLORIDE

 DIALKYL DIMETHYL AMMONIUM BROMIDE
     100Z-C12     (Use for.dicoco-)

 DIALKYL DIMETHYL AMMONIUM CHLORIDE
     1001-C12     (Use for dicoco-)
       4X-C14, 26X-C16, 70VC18

 Dl-n-ALKYL METHYL BENZYL AMMONIUM CHLORIDE
     100I-C12
       5X-C12, 60Z-C14, 30VC16, 5I-C18  (BTC 776)

 DI(ALKYL OXYPROPYL) DIMETHYL AMMONIUM CHLORIDE
     100I-C10
      60Z-C8, 407o-ClO        (Q-Dox)

p-DIISOBUTYLCRESOXYETHOXYETHYL DIMETHYL BENZYL
           AMMONIUM CHLORIDE
                monohydrate

 p-DIISOBUTYLPHBNOXYETHOXYETHYL DIMETHYL BENZYL
           AMMONIUM CHLORIDE
                monohydrate
                                                                         M.W.    ^HALOGEN(FACTOR)   I N  (FACTOR)
C19H42NC1

C23H41N2OC1
C H NBr
^ A, *f \J
C21H38NC1

C5H13C1NC1
C26H56NBr
C26H56NC1
fc W J U
C32H60NCl


C H 0 NCI

C9HH..O,NC1
Zo 44 2

C, H, 0 NCI




320
346
397
384
402
340
358
158
462
416
567
494
570

478
444
462
480

448
466



.01
.7
.05
.45
.47
.00
.01
.07
.65
.19
.4
.29
.0

.25
.6
.12
.14

.09
.11



11
10
8.
20
19
10
9.
22
17
8.
6.
7.
6.

7.
7.
7.
7.

7.
7.



.08
.23
929
.78
.85
.43
903
.43
.27
478
248
172
219

413
974
672
384

912
593



7.C1(9.026)
7oCl(9.
%C1(11
7.Br(4.
778)
.20)
811)
2Br(5.037)
7.C1(9.
7.C1(10
590)
.10)
7.C1(4.459)*
7.Br(5.
7.C1(11
%C1(16
7.C1(13
7.C1(16

7.C1(13
%C1(12
7.C1(13
7.C1(13

7.C1(12
7.C1(13



790)
.80)
.01)
.94)
.08)

.49)
.54)
.03)
.54)

.64)
 17)



4
4
3
3
3
4
3
8
3
3
2
2
2

2
3
3
2

3
3



.377
.041
.528
.643
.480
.120
.912
.861
.028
.349
.468
.834
.457

.929
.151
.031
.917

.126
.000



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.85)
.75)
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.45)
.73)
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.56)
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.86)
.51)
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.70)

.14)
.74)
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.28)

.99)
.33)












sj


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a
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3
M
H"

rt
o
o
C
A
O.
to
                                                                                                                    w

-------
DIQUAT DIBROMIDE
        monohydrate

DODECYLACETAMIDYL DIMETHYL BENZYL AMMONIUM  CHLORIDE
              (NOPCO, DBC)

DODECYLBENZYL TRIMETHYL AMMONIUM CHLORIDE
              (Barquat, TC-50, DBQ, GT-50,  LQ-150)

DODECYLBENZYL TRIMETHYL AMMONIUM 2-ETHYLHEXOATE

DODECYL DIMETHYL BENZYL AMMONIUM CYCLOPENTANE
                CARBOXYLATE  SALT

FURFURYL TRIMETHYL AMHDWIUM  IODIDE

2-HEPTADECENYL-l-ETHANOL-l-ETHYL IMIDAZOLINIUM BROMIDE

2-HEPTADECYL-l-METHYL-l-(2-(STEAROYLAMIDO)ETHYL)  IMID-
              AZOLINIUM METHYL SULFATE    (Arqual  S)

1,3-bis(2-HYDROXYETHYL)-2-HEPTADECENYL  IMIDAZOL-
                  INIUM CHLORIDE

1,3-bi3(2-HYDROXYETHYL)-2-HEPTADECENYL  IMIDAZOL-
                  INIUM BROMIDE

METHYLALKYLBENZYL TRIMETHYL  AMMONIUM  CHLORIDE
    1007.-C12

METHYLDODECYLBENZYL TRIMETHYL AMMONIUM  CHLORIDE

METHYLDODECYLXYLYLENE bis(TRIMETHYL AMMONIUM CHLORIDE)

METHYLDODECYLBENZYL TRIMETHYL AMMONIUM  CHLORIDE( 807.)
       METHYLDODECYLXYLYLENE bis(TRIMETHYL  AMMON-
         IUM CHLORIDE) (207.)          (Hyamine  2389)
C23H41N2OC1
C22H40NC1
C30H55N02
C27H47N02

C.H,,NOI
 O 14

C24H47N2Br
C42H85N35S
C9/H. N 0 Br
 24 47 2 2
C23I!42NC1
                 M.W.    7.HALOGEN(FACTOR)   7. N  (FACTOR)
                344.06   46.45 7.Br(2.153)  4.071    (24.56)'
                362.08   44.14 7.Br(2.266)  3.868    (25.85)'
397.05   8.929 70C1(11.20)   3.528    (28.35)'


354.02   10.01 7.C1(9.986)  3.957    (25.27)

461.78                     3.033    (32.97)


417.68                     3.354    (29.82)

267.11   47.51 7.1 (2.105)  5.244    (19.07)

459.76   17.39 7.Br(5.751)   3.047    (32.82)'


744.23   4.3087.3(23.21)   1.882    (53.13)'
431.11


475.56


368.05

368.05

475.63
                                                             00
                         8.224 XC1(12. 16)   3.249    (30.78)'
16.80 7.Br(5. 951)   2.945    (33.95)


9.633 7.C1(10.38)  3.806   (26.28)

9,.633 7.C1(10.38)  3.806   (26.28)

14.91 7.C1(6.708)   2.945   (33.96)1
                                                             9
                                                           '
                                                             n
                                                             n
                                                             g
                                                             i
                331.7    10.69 7.C1(9.356)  4.223   (23.68)   o
                                                                                                                     a
                                                                                                                     Q.
                                                                                                                     n
                                                                                                                     w

-------
OCTADECYL TRIMETHYL AMMONIUM  PENTACHLOROPHENATE



PARAQUAT D1CHLORIDE (1,1'-D1METHYL-4,4'-BIPYR1D-

           INIUM DICHLORIDE)



PARAQUAT Dl OR bta METHYL  SULFATE  (1,1'-DIMETHYL-

        4,4'-BIPYRIDlNlUM  DIMETHYL SULFATE)



TRIMETHYL OCTAOECENYL AMMONIUM CHLORIDE



TRIMETHYL OCTADECADIENYL AMMONIUM  CHLORIDE
C14H208N2S2
                 M.W.    7J1ALOGEN(FACTOR)   7. N  (FACTOR)



                577.9                      2.424 %N(41.26)
C12H14N2C12     257.16   27.57 %C1(3.627)  5.447    (18.36)'
                     6   15.707.3(6.369)  3.429   (29.16)'



                346.04   10.25 7.C1(9.761)  4.048 7.N(24.71)



                344.03   10.31 7.C1(9.704)  4.071 7.N(24.56)
                                                                                                                    .C
                                                                                                                     rt>
                                                                                                                     n
                                                                                                                     a
                                                                                                                     o
                                                                                                                     a
                                                                                                                     n
                                                                                                                     o
                                                                                                                     s
                                                                                                                    o
                                                                                                                     o
                                                                                                                     c

                                                                                                                     O-
                                                                                                                     w

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January 1976                                  Quaternary Ammonium Compounds



          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 mg 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 Ferricyahide 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 ferricyanide 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:
     %N
(Blank ml - Sample ml)(N Na,,S203) (.0140) (100)
                            (grams sample)




    . 0.01AO = 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
                       + K,Fe(GN), - } 3KX + [R-R.R-R. ].Fe(CN),4,
                          J      b               1 Z J 4 j      o
    2.  Reaction of excess ferricyanide with potassium iodide
         Excess 2K,Fe(CN), + 2KI - \ 2K.Fe(CN), + I_
                  JO            '   4      D    i
3.  Removal of K.Fe(CN), by zinc sulfate to speed oxidation of
                                                                   KI
         2K4Fe(CN)6 + 3ZnS04
    4.  Titration of released iodine by sodium thiosulfate
         I2 + 2Na2S203 - ) Na2S46 + 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 mi 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 solution 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 Nitroeen = (Blank ml - Sample ml)(M)(.QIAO)(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

        Determination of Quaternary Chlorides and Bromides
   in Mixed Quaternary Formulations by Poteritiometric 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 Vplhard titration).  Place the;
     electrodes in the solution and set the potential on the titrimeter
     at 0.7 or 0.8 volt.  Add 0.1N 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 AgNO.)(.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

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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, 30% cis isomers),  a registered insecticide having the
chemical structure:
     CH2I
                                          0
                           CH20CCH
XCH3

\H3
   C^*i i ^^^~ ^*    ^*\ i
CHCCH3
Molecular formula:  C  H  0
                     2.L Zo J
Molecular weight:   338
Melting point:      A3 to A8C
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-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 rag/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 35-40C.  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 jj) and baseline from 1765 cm"  to
     1660 cm'1 (5.67 ji to 6.02 >i).

     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)(2 purity std)
         (abs. std)(cone, sample in mg/ml)
Method submitted by Mark Law and Jack Looker, EPA Beltsville Chemistry
Laboratory, TSD, OFF, Beltsville, Maryland.

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  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:
                                            0
                                            II
                             -CH20CCH
                                                      XCH=CCH3
-j
  Molecular  formula:  C00H0,0_
                       it.  /O J
  Molecular  weight:   338
                                              a
  Melting  point:      43 to 48C
  Physical state,  color, and odor:  waxy off-white to tan solid with a
                     characteristic chrysantheraate 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.

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                                 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:     260C
         Injection temperature:  290C
         Detector temperature:   270C
         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
           j&
     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
     35-40C.  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 pi 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:


      m (pk. ht. or area sample)(wt. std injected)(% 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.

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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-
diraethyl-3-.(2-methylpropenyl) cyclopropanecarboxylate  (approx. 70%
trans, 30% cis isomers), a registered insecticide having the
chemical structure:
                                                        /CH3
                                           0
                                           II
                              CHg0CCH
                                                                 CH3
Molecular formula:  C_-H0,0_
                     zz /o j
Molecular weight:   338
Melting point:      43 to 48C
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:     230C
     Injection temperature:  250C
     Detector temperature:   250C
     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 ml 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 propellants1 by placing can in a water bath at about
     35-40C.  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)(100)
         (wt. sample)(pk. ht. or area dieldrin)(RF)
Method submitted by Stelios Gerazounis, EPA Region II, New York, New York.

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 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:
                                           0
                                                       X
                                                          CH3
\- CH2 
-/
                              CH20CCH
C CH=C CH3
  Molecular  formula:  C_0H.,0.
                      a /o j
  Molecular  weight:   338
  Melting  point:      43 to 48C
  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.

-------
                                                      Resmethrin EPA-4
                                                      (Tentative)
Reagents;
     1.  Resmethrin standard of known % purity
     2.  Methanol, pesticide or spectro grade

Equipmenti
     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:  65C
     Column pressure:     1000 psi
     Observed flow rate:  1-2 ml/min
     Detector:            UV at 254 nm
     Chart speed:         5 min/in
     Injection:           5 jil

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 propellants by placing can in a water bath at
     about 35-40C.  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 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 resmethrin 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 Hayes, EPA-OPP-TSD Beltsville Chemistry
Laboratory, Beltsville, Maryland.

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October 1975
Resmethrin EPA-5
(Tentative)
                  Determination of  Resmethrin by
                    Gas-Liquid  Chroraatography
                    (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:
                                           0
                                                       X
                              CH20CCH
                                                     *CCH=CCH3
                                                                 CH3
Molecular formula:  C__H0,0_
                     Li 2o J
Molecular weight:   338
Melting point:      43 to 48C
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 restnethrin 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)
      I
         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)(100)
         (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:
                                        ci
         CH30
Molecular formula:  C_H~C1_O.PS
                     o o  j j
Molecular weight:   321.5
Melting point:      softens at 35 to 37C with full melt at 40-42C
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 60C, 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-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~  to 890 cm   (9.8 u to  11.3 u).
         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 fi).

     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.

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Navember 1975
Ronnel EPA-2
                     Detennination 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:
           CH3	0
           CH3	0'
                           po
Molecular formula:  C-H_C1-O.PS
                     o o  J J
Molecular weight:   321.5
Melting point:      softens at 35 to 37C with full melt at  40-42C
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 60C, 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 pi
     4.  Mechanical shaker
     5.  Centrifuge or filtration equipment
     6.  Usual laboratory glassware

Operating Conditions for FID;
     Column temperature:     180C
     Injection temperature:  250C
     Detector temperature:   250C
     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 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 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


     pp a (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:


     %  (wt. I.S.)(% purity I.S.)(pk_. ht. or area Ronnel) (100)
         (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 A0506.

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January 1976
Rotenone EPA-1
                    Determination of Rotenone
                in Pesticides - Qualitative tests

     Rotenone is a registered insecticide having the chemical structure:
                     0-CH3
                                0
Molecular formula:  C00H._0
Molecular weight:   394.4
Melting point:      163C (a dimorphoric form melts at 181C)
Physical state and color:  colorless crystals; crystallizes with solvent
                    of crystallization
Solubility:  15 ppm in water at 100C; 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-l









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.

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November 1975
Salicylanillde EPA-1
                 Determination of Salicylanilide
                   by Ultraviolet Spectroscopy

     Salicylanilide is a registered fungicide having the chemical
structure:
                         0     H
Molecular formula:  C  H  NO
Molecular weight:   213.3
Melting point:      135C
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:
     Pjrejparatiqn of Standard:
         Weigh 0.1 gram salicylanilide into a 100 ml volumetric flask;
     dissolve in  and make to volume with 0.1N 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 jig/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 jag 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 0.1N sodium hydroxide
solution, and mix thoroughly.  Pipette 10 ml into a second 100 ml
volumetric flask; make to volume with the 0.1N sodium hydroxide
solution.  (final cone 10 ^ig salicylanilide/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 338 nm with 0.1N
sodium hydroxide solution in each cell.  Scan both the standard and
sample from 360 nm to 250 nm with 0.1N 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 blank.  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:

7 - (ahs. sample)(cone, std in pg/ml)(% purity std)
    (abs. std) (cone, sample in jag/ml)

-------
September 1975                                          Siduron EPA-1

                                                        (Tentative)


                     Determination of  Siduron

                   by Ultraviolet  Spectroscopy



     Siduron is the accepted  common name  for l-(2-methylcyclohexyl)-


3-phenylurea, a registered herbicide having the chemical structure:
                                          CH	CH2
                         H   0   H     /          \
                  xx    I    II   I     /            \
                   yNCNCH              CH2
                                                CH2
Molecular formula:  Ci^on1*?0


Molecular weight:   232.3


Melting point:      133 to 138C


Physical state, color, and odor:  odorless, white, crystalline solid


Solubility:  18 ppm in water at 25C;  soluble  to  the extent of 10% or

             more in cellosolve, dimethylacetamide, dimethylformamide,

             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 run with

-------
                                                         Siduron  EPA-1
                                                         (Tentative)
     methanol in each cell.  Scan both the standard and sample from
     300 run to 200 run 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:


      m (abs. sample)(cone, std in ;ig/ml)(% purity std)
         (abs. std) (cone, sample in jjg/ml)
Method submitted by Stelios Gerazounis, EPA Region II, New York, N. Y.

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September 1975
                Slmazine  EPA-1
                (Tentative)
            Determination of  Slmazine In 0.1% Aqueous
              Suspension by Ultraviolet Spectroscopy
     Simazine is the accepted  common name for 2-chloro-4,6-bls
(ethylamino)-s-triazine,  a registered herbicide having the
chemical structure:
                               Cl
                        N
 CH3CH2N	C
                                            H
CNCH2CH3
                               N'
Molecular formula:  C?H -C1N
Molecular weight:   201.7
Melting point:      225 to 227C
Physical state and color:  white,  crystalline solid
Solubility:  at 20C, 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

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                                 2                        Simazine 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 mu
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
     A.  Usual laboratory glassware

Procedure;
     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 ug/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

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                                 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 jig simazine/ml)

     UV Determination;
         Using the optimum quantitative settings for the particular
     UV instrument being used, adjust the 0 and 100% settings at
     263 mu with methanol in both cells.  Scan both standard and
     sample from 360 mu 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:
         ^A263~A300 samPle)(conc* std in jig/ml) (% purity std)
       =            stdHconc. sample in ;ig/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.

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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:
Na
                               0=CI=0
                                      II
                                      0
Molecular formula:  NaCIO
Molecular weight:   106.4
Melting point:      248C; decomposes about 300C 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 0C and 230 g/100  ml at
             100C; 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.

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                                 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,.4H.O) into 200 ml volumetric flask, add 25 ml
         sulfuric acid and 25 ml 85% 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
                             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.  Mechanical shaker
     3.  Filtration apparatus
     4.  Titration apparatus
     5.  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
     ./ x, oi r,    (net ml KMnO.)(N KMnO.)(0.01774)(100)
     % NaC10_ = 	4	4	

                       (gram sample)(25/250)
     0.01774 - milliequivalent weight of NaC103

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   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
H2NC
        Streptomycin  is  a  strongly basic compound with the empirical


   formula  C2iH39N7i2'  mlecular 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:   ^C2lHi9N7i2^2"3H2S4


  (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

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                                 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, streptomycine (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 0.48 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 O.A8 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 nm
     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,

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                            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 A50 run 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

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December 1975
         Strychnine EPA-1
                   Determination of Strychnine
          in Poisoned  Baits  (Picric acid precipitation)
     Strychnine is a registered rodenticide having the chemical
structure:
                          yv          i b^ **x
                                              ,o
                  >CH         N
            CH     XC^
            CH         C	^
                 JCH          CH2
                              CH2-
CH
N
II
C
Molecular formula:   C9iH22N202
Molecular weight:    334.4
Melting point:      268  to  290C  (depending on the speed of heating)
                    with decomposition; b.p. 270C 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

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                                 2                     Strychnine EPA-1





     Strychnine generally is used as the sulfate; poison baits usually




are colored grain containing 0.5 to 1% strychnine sulfate.




     Strychnine sulfate is a white crystalline powder containing 5




moles of water of crystallization lost at 110C; moderately soluble




in water and alcohol, insoluble in ether; mol. formula:  (C-.H  N 0 ) -




H SO .5H 0; mol. wt. 856.96; m.p. above 199C.






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;




     1.  Ether-chloroform mixture (2 parts ethanol + 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.  Pour 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

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                                 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 105C and weigh.
Calculations:
                       (grams strychnine picrate) (0.5934) (100)
                          gample)(100/150)(x/10o)(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

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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:  C2iH22N22
Molecular weight:   334.A
Melting point:      268 to 290C (depending on the speed of heating)
                    with decomposition; b.p. 270C 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 preclpltants
             (e.g., picric acid as in this method)
Other names: Kwik-kil, Mouse-tox, Ro-Dec

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                                 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 run 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

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                                 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 with the 0.5% 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
     0.5% sulfuric acid.  Mix each flask thoroughly,  (final cones
     7.5, 15.0, and 22.5 pg/ml)
         (If a direct standard - sample comparison is to be made,
          use 15 jig/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,

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                            4                      Strychnine EPA-2

draining each extract through plug of cotton (prewashed with chloro-
form) into 400 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 40 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 0.5% 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 nra 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  Cabs, 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:
     v a (abs. spl)(conc. std in ug/ml)(% purity std)
         (abs. stand.)(cone, sample in pg/ml)

     or using dilution factors:
     .   (abs. Spl)(&   std)(purity std)(1/100)(15/100)(10/100)(100)
     7a " (abs. stand.) (g   sample) (1/100) (25/100) (25/100)

Method developed by Lawrence A. Wapensky (Journal of the AOAC, Vol. 52,
No. 5, 1969, pages 1015-1016).

(The format of the method has been changed somewhat to conform to the
 general format as used for the methods in this manual.)

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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:  115C; b.p. 444.6C
Physical state and color:  yellow solid, melting at 115C to a yellow
                mobile liquid which darkens and becomes viscous about
                160C.  It exists in two allotropic forms:  rhombic,
                m.p. 112.8C, and monoclinic, m.p. 119C.
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 (100-105C)
     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
100-105C and weigh.  Subtract to determine the weight of elemental
sulfur.
     Using the above weight, calculate the percent sulfur in the sample
as follows:

         v  ul   . (vt. elemental sulfur) (100)
                           (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.

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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:  115C; b.p. 444.6C
Physical state and color:  yellow solid, melting at 115C to a yellow
                mobile liquid which darkens and becomes viscous about
                160C.  It exists in two allotropic forms:  rhombic,
                m.p. 112.8C, and monoclinic, m.p. 119C.
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

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                                                          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.

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                                 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 550-650C.  Weigh as



barium sulfate.



     Calculate the percent sulfur in the sample as follows:



         .    .     ,,      (wt. of precipitate)(100)
         % Barium sulfate 	K	 v ,^, \^
                                (wt. sample)(50/250)






         % Sulfur - (0.1374)(% barium sulfate)

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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:  115C; b.p. 444.6C
Physical state and color:  yellow solid, melting at 115C to a yellow
                mobile liquid which darkens and becomes viscous about
                160C.  It exists in two allotropic forms:  rhombic,
                m.p. 112.8C, and monoclinic, m.p. 119C.
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

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                                 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 ml Erlenmeyer flask (preferably with pour-




out lip), add 50 ml of the sulfur-saturated acetone, stoppered tightly,




and shake for several minutes 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 disulfide 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




105C for 15 minutes.  CAUTION - carbon disulfide is extremely flammable!

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                                 3                         Sulfur EPA-3




     Cool, weigh, and calculate the percent carbon disulfide soluble


sulfur as follows:



         X sulfur - <"t- residue) (1-001
                      (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
                                                         \r

       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 105C 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.

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December 1975                                         Sulfur Dioxide EPA-1

                 Determination of Sulfur Dioxide
                    in Fumigants by lodometry

     Sulfur dioxide is a registered fumigant, having the chemical
structure:
Molecular formula:  SCL
Molecular weight:   64.07
Boiling point:     -10C
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

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                                 2                       Sulfur Dioxide EPA-1

Principle of 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 tltration.

Calculation;
     Calculate the sulfur dioxide as follows:

     % a [(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 Na0S00_)(N Na0S.O.)(0.03203)(100)
                         (wt. sample in grams)

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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:
                                                      H
                                          0  CNCH3
                            CH3-C-CH3
Molecular formula:  C -H.-NO
Molecular weight:   277.4
Melting point:      200 to 201C;  the technical product is 95% and has
                    a mp of 185 to 190C
Physical state, color, and odor:   white,  odorless,  crystalline solid
Solubility:  7 ppm in water at 25C;  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

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                                 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)

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                                                          Terbutol EPA-1
                                                          (Tentative)
     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
     1580 cm"1 (5.2 ji to 6.3 ;i).
         Determine the absorbance of standard and sample using the peak
         754 cm"1 (5.7 ;
     (5.45 p to 5.9 ;i).

     Calculation:
at 1754 cm"  (5.7 ji) 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:

     y  (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.

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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:
            CH-
                                                0    H
0CNCH3
                            CH.-C-CH3
Molecular formula:  C 7H  NO
Molecular weight:   277.4
Melting point:      200 to 201C;  the  technical  product is 95% and has
                    a mp of 185 to 190C
Physical state, color, and odor:   white,  odorless, crystalline solid
Solubility:  7 ppm in water at 25C;  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

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                                 2                         Terbutol EPA-2
                                                           (Tentative)
Reagents;
     1.  Terbutol standard of known % purity
     2.  Diazinon standard of known % purity
     3.  Acetone, pesticide or spectro grade
     4.  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 jil
     4.  Mechanical shaker
     5.  Centrifuge or filtration equipment
     6.  Usual laboratory glassware

Operating Conditions for FID;
     Column temperature:     16SC
     Injection temperature:  215C
     Detector temperature:   215C
     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.

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                                                         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 3d minutes
     or shake by hand intermittently for one hour (final cone 2 mg
     terbutol and 2 mg diazinon/ul)
                                        \
     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 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.

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                                                        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)(100)
         (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 Chemistry Lab.  Any comments, criticisms,

       suggestions, data, etc. concerning this method will be appreciated.

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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:
      CH3             I                 S          .CH3
                  	C	SSCN
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                                 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 jig 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

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                                                      Thirara EPA-1
chloroform in each cell.  Scan both the standard and sample from
350 nm to 250 nm with chloroform In the reference cell.
    Measure the absorbance of standard and sample at 280 nm.

Calculation!
    From the above absorbances and using the standard and sample
concentrations, calculate the percent thlram as follows:

 m (aba, sample)(cone, std in jug/ml) (% purity std)
    (abs. std)(coric. sample in jig/ml)

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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:
      CH3
Molecular formula:  C-H, _N.S.
                     6 12 2 4
Molecular weight:   240.44
Melting point:      155 to 156C
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

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                                 2                         Thiram  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 0.065 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.

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                                 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)

     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 standard and sample from 1430 cm   to
     1300 cm'1 (7 ji to 7.7 ;i).
         Determine the absorbance of standard and sample using the
     peak at 1380 cm   (7.25 ju) and baseline from 1400 cm"  to 1350 cm"
     (7.14 p to 7.41 ji).

     Calculation:
         From the above absorbances and using the standard and sample
     concentrations, calculate the percent thiram as follows:
      _ (abs. sample)(cone, std in mg/ml)(% purity std)
         (abs. std)(cone, sample in mg/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.)

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January 1976
                        Trichlorocarbanili.de  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:
  o-
  Cl	1
H     0    H
 I     II      1
NC	N
Molecular formula:  C -H Cl N 0
Molecular weight:   315.6
Melting point:      250C (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

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                                 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 Sample:
         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  ig trichlorocarbanilide/ml)

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                                               Trichlorocarbanilide 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 265 hra 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:

7 - (abs. sample) (cone, std injug/ml)(% purity std)
    (abs. std)(cone, sample in jug/ml)

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November 1975
Trifluralin EPA-1
                   Determination of Trifluralin by
                    Gas-Liquid  Chromatography
                    (FID - Internal Standard)

     Trifluralin is the accepted common name for a,a,cy-trifluoro-2,6-
dinitro - N,N-dipropyl-p-toluidine, a registered herbicide having the
chemical structure:
        CH3 CH2CH2NCH2CH2 CH3
Molecular formula:  C,,H1/:F,N_0.
                     1J lo 3 3 4
Molecular weight:   335.3
Melting point:      48.5 to 49.0C  (tech. product is at least 95% pure
                                    and has a mp greater than A2C)
Physical state, color,  and odor:,  orange crystalline solid; no
                    appreciable odor
Solubility:  less than 1 ppm in water  at 278C; 7% in ethanol, 40% in
             acetone, 58% in xylene; soluble in other organic solvents
Stability:   stable but susceptible to photochemical decomposition

Other names: Treflan (Ell Lilly), Trefanocide, Treficon, Triflurex,
             Su Seguro Carpidor

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                                 2                       Trifluralin EPA-1
Reagents;
     1.  Trifluralin standard of known % purity
     2.  Diisobutylphthalate
     3.  Acetone, pesticide or spectro grade
     A.  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 dlisobutylphthalate/ml)

Equipment:
     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 jul
     4.  Mechanical shaker
     5.  Centrifuge or filtration equipment
     6.  Usual laboratory glassware

Operating Conditions for FID;
     Column temperature:     200C
     Injection temperature:  210C
     Detector temperature:   275C
     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-1
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 mg 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 rag
     trifluralin and 12 mg diisobutylphthalate/ml)

     Determination:
         Inject 2-3 /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 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.

-------
                                 4                     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

     RF a (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:

     7 = (wt. I.S.)(% purity I.S.)(pk. ht. or area trifluralin)(100)
         (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,a,o?-trifluoro-
2,6-dinitro-N,N-dipropyl-p-toluidine, a registered herbicide
having the chemical structure:
       CH3CH2CH2 NCH2CH2CH3
Molecular formula:  C._H,-F-N.O.
                     1J lo J j  H
Molecular weight:   335.3
Melting point:      48.5 to 49.0C  (tech. product is at least 95% pure
                                   and has" a mp greater than 42C)
Physical state, color,  and odor:  orange crystalline solid; no
                    appreciable odor
Solubility:  less than 1 ppm in water at 27C; 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

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                                 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-mg/ml)

-------
                                                    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 trifluralin/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  4 mg trifluralin/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 1390 cm
to 1212 cm"1 (7.2 p to 8.25 ;).
    Determine the absorbance of standard and sample using the
peak at 1300 cm"  (7.69 >i) and baseline from 1315 cm   to 1264 cm
(7.6   to 7.91 p).

-------
                                                        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 absorbarice of approx. 0.079 in a 0.2 mm cell.)
Method contributed by the Commonwealth of Virginia, Division of
Consolidated Laboratory Services.

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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:


                               ?          ,CH2CH2CH3
CH3CH2CH2SC	N<^
                                           XH2 CH2CH3

Molecular formula:  C  H  NOS
Molecular weight:   203.4
Boiling point:      140C at 20 mm Hg, 150C at 30 mm Hg
Physical state, color, and odor:  clear liquid  with  an aromatic odor
Solubility:  about 100 ppm in water at 20-21C; 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 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 0.12 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 1R instrument
being used, scan both the standard and sample from 1850 cm   to
1500 cm'1 (5.4 /i to 6.7 ^).
    Determine the absorbance of standard and sample using the
peak at 1630 cm~  (6.13 p) and basepoint at 1800 cm~  (5.56 ju) .

Calculations;
    From the above absorbances and using the standard and sample
concentrations, calculate the percent vernolate as follows:

"i _ (abs. sample) (cone, std in mg/ml) (% purity std)
0   (abs. std)(cone, sample in mg/ml)
(A concentration of 1 mg vernolate/ml chloroform gives an
 absorbance of approx. 0.02A 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 10.5/0




     Analytical peak:  1105 cm   (9.05 fi)




     Baseline:   1163 cm"1 to 1047 cm"1 (8.6 ^i 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:
                                            CH2CH2CH3
CH3CH2CH2SC	N
                                                  CH2CH3
  Molecular  formula:  C  H  NOS
  Molecular  weight:   203.4
  Boiling  point:      1AOC at 20 mm Hg, 150C at 30 mm Hg
  Physical state, color, and odor:  clear liquid with an aromatic odor
  Solubility:  about 100 ppm in water at 20-21C; 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 pi
     4.  Mechanical shaker
     5.  Centrifuge of filtration equipment
     6.  Usual laboratory glassware

Operating Conditions for FID;
     Column temperature:     140C
     Injection temperature:  225C
     Detector temperature:   250C
     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 rag 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.

-------
                                                            Vernolate EPA-2
          Determine the RF value for each injection of the standard-
     internal standard solution as follows and calculate the average:
     RF
(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 -    ;. cycloate) (% purity cycloate) (pk. ht. or area vernolate) (100)
         (wt. sample) (pk. ht. or area cycloate) (RF)
Method submitted by Division of Regulatory Services, Kentucky
                        >              .
Agricultural Experiment Station, University of Kentucky, Lexington,
Kentucky 40506.

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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
                                             CH2CH2CH3
 CH3CH2CH2SC
                                                 CH2CH3
Molecular formula:  C  H  NOS

Molecular weight:   203.4

Boiling point:      140C at 20 mm Hg,  150C at 30 mm Hg

Physical state,  color, and odor:  clear liquid with an aromatic odor

Solubility:   about 100 ppm in water at  20-21C; miscible with common
             organic solvents

Stability:   stable; non-corrosive


Other names:  Vernam (Stauffer), R-1607, S-propyl M,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 + 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 ^il
     4.  Mechanical shaker
     5.  Usual laboratory glassware

Operating Conditions for TCP:
     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 jal 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)
     RF = (wt. butylate)(% purity butylate)(pk. ht. or area vernolatel
          (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 = (yt. butylate)(% purity butylate)(pk.  ht. or  area vernolate)(100)
         (wt. sample)(pk. ht. or area butylate)(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,

-------
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:
                                          c=o
Molecular formula:  C-_H..,0,
Molecular weight:   308.3
Melting point:      (dl form) 159 to 161eC
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 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:        10% methanol + 90% 0.0025M H_PO. in water
                                                      34
     Column temperature:  50C
     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 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
     warfarin as follows:
         (pk. ht.
         (pk. ht.
or area sample)(wt. std injected)(% purity of std)
or area standard)(wt. sample injected)
Method submitted by Elmer H. Hayes, EPA, Beltsville, Md.

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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 :
                                          c=o
Molecular formula:  C,_H,,0,
                     19 16 4
Molecular weight:   308.3
Melting point:      (dl form) 159 to 161C
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.P 0 .10H 0 in water and make to 500 ml.
     3.  Ether-hexane mixture - extract 200 ml n-hexane (bp 6068C)
         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 glassx^are

Procedure:
     Preparation 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

-------
                            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, .centrif.uge 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 |ig warfarin/ml)

UV Determination:
    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.  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 warfarin as follows:

, _ (abs. samj?j.e) (cone. std in pg/ml) (% purity std)	
    (abs. std)(cone, sample in ug/ml)

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December 1975
Warfarin EPA-3
(Tentative)
              Determination of Warfarin, Sodium Salt
              by High Pressure Liquid Chroraatography

     Warfarin is the official common name for 3-(alpha-acetonylbenzyl)-
4-hydroxycoumarin, a registered rodenticide having the chemical struc-
ture:
                           OH
Molecular formula:
Molecular weight:   308.3
Melting point:      (dl form) 159 to 161C
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

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                                 2                       Warfarin EPA-3
                                                         (Tentative)
Reagents;
     1.  Warfarin standard of known % purity
     2.  Methanol, pesticide or spectro grade
     3.  Phosphorous acid solution, 0.0025M in water
     4.  Sodium pyrophosphate, 1% solution - dissolve 10 grams
         Na.P.O-.lOH 0 in water and make to 1000 ml.

Equipment;
     1.  High pressure liquid chromatograph with UV detector at 254 tun.
         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 run.
     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.PO. in water
                                                      34
     Column temperature:  50C
     Chart speed:         5 min/inch or equivalent
     Flow rate:           0.5 to 1.5 ml/min (Perkin-Elmer 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 reproduclbility.

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                                 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 rag/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
     25A nm.  Warfarin is more easily detected at this wavelength and
     many interferences are eliminated or reduced to a negligible
     amount.
         Alternately inject three 5 jul 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:
     j e (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.

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December 1975
Zinc Phosphide EPA-1
                 Determination of Zinc Phosphide
                by the Phosphlne Evolution Method

     Zinc phosphide is a registered rodenticide having the chemical
structure:
Molecular formula:  Zn->po
Molecular weight:   258.1
Melting point:      420C (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
                                                                           XYZ

-------
                                 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
         KMnO^, 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 15C
     5.  Nitrogen gas
     6.  Sodium bicarbonate, saturated solution
     7.  Starch indicator solution
     8.  Iodine, 0.1N standard solution

Equipment;
     1.  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
     2.  Water bath maintained at 50C
     3.  Titrating equipment
     4.  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:

    % o (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 50C
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 50C 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 + ml used  in titration)



meg's from oxalic acid a N x ml oxalic acid used	



meq's difference  net meq's used by sample
-  4    ,    U4,     (net meq's) (0.01613) (100)
% zinc phosphide 	7-1*-*-.	r-^r	
       f                (grams  of sample)


           i

                                                         (258  09)
milliequivalent weight of zinc phosphide  is 0.01613   or  V. ,>/inoo\
Reactions;
            (COOH)2
     (COOH)2 + 2KMn04 +  3H2S04 - )  ^2*^  + 2MnS4 + 10 C2^ + 8H2

-------
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_P.
Molecular weight:   258.1
Melting point:      420C (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 phoaphine

Other names: Kilrat, Mous-con, Rumetan
Reagents:
     1.  Zinc phosphide standard of known % purity
     2.  Glucose, 100 mesh, dry powder
     3.  Toluene, pesticide or spectro 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 40-50C (or equivalent column)
     3.  Precision liquid syringe:  10 ul
     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:     40-50C
     Injection temperature:  200C
     Detector temperature:   140-150C
     Nitrogen carrier gas:   45-60 ml/min
     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 than 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 achieved by 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 jig PH /ml)

      (0.3794 g of the 1% mixture will hydrolyze to 1 mg phosphine,
       which, dissolved in 100 ml toluene, gives a 10 ug/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:

        (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.

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                                                   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, with
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 100C 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 -105C 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 110C 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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                                             TLC System 1
Acceptable
     Aluminum Oxide G and Silver Nitrate Layer

Spots Visualized by Exposure to UV Lamp (unfiltered)


                                              2/
                               Mobile Solvents^'
Name
(Herbicides)
Lasso
Tordon, Methyl
Ester
Aminotriazole
Atrazine
Barban
Prometone
Prometryne
Bromacil
Chlorbromuron
Propazine
Chloroxuron
Dursban
Dalapon
No.
11
39A
40
63
68
96
97
111
17 3A
184
217B
219AA
273
Hexane Benzene

0 .14
0 0 	 .13
0 0
0 .04
0 0 	 .28
No data (ND) ND
ND ND
0 0
0 .23
0 .08
0 .03
0 0 	 .07
0 0
Cy c lohexane+
Benzene+HAC+
Paraffin Oil
Chloroform (200+30+20+11)
3/
.47 0, .66
0 	 .35 0, .13
0 0
.23 0, .60
0 	 .50 0, .35
ND 0 	 .53
ND 0 	 .59
.02 0, .45
.57 .40
.33 .70
.22 .14
0 0 	 .33
0 0
Ethyl
Acetate

.72
0, .63
0
.60
0, .73
ND
ND
.02 	 .07
.72
.60, .70
.63
0 	 .08
0

-------
Acceptable
Mobile Solvents^'
Name
(Herbicides)
Banvel D
2,4-D Acid
2,4-D Butyl
Ester
2,4-D Ethyl
Ester
2,4-D Ethyl
Hexyl
2,4-D IOE
2,4-D Isbpropyl
2,4-D Prop. Gly.
But . Ether
2,4-DB Acid
2,4-DB IOE
TDK
Stam
Dacthal
No.
295
315
315
AL
315
AP
315
AS
315
AU
315
AV
315
BA
316
316D
323D
325
382
Hexane

0
0
0
0
0
- 0
0
0
0
0
ND
ND
0
Benzene Chloroform
RValitoe- 
0 0
0 0
0, .57 0, .72
0 	 .53 0, .70
0, .65 0, .77
0, .60 0, .75
0, .55 0, .75
0, .45 .72
0 0
0, .62 .83
.67 .77
.12 .42
60 .75
Cyclohexane+
Benzene+HAC+
Paraffin Oil
(200+30+20+11)

o  !os
0 	 .02
0, .73
0, .72
0, .80
ND
0, .75
0 , . 75
0 	 .30
.85
.80
.07
.80
Ethyl
Acetate

0
0
0, .77
0, .75
0, .80
0, .77
0, .80
0, .77
0
.79
.82
.67
^-*
                                                                                                      .77

-------
Acceptable
               21
Mobile Solvents
Name
(Herbicides)
Diuron
Linuron
MCPA
MCPA dimethyl
amine
MCPA IOE
MCPB
MCPP
Monuron
Monuron TCA
Neburon
Paraquat
Fenuron TCA
Tordon
Siduron
Silvex
Silvex IOE
No.
410
528
557C
557G
5571
558
559
583
583A
594
634
655
663AA
733A
739
7391
Hexane

0
ND
0
ND
ND
ND
ND
0
ND
ND
0
ND
0
0
0
ND
Cyc lohexane+
Benzene+HAC+
Paraffin Oil
Benzene Chloroform (200+30+20+11)
RValnoo. ,_j 	 .,_ ju^ _n_ ._ ..,_ j_ _.
0 .27 0 	 .03
.25 .53 0, .55
0 0 0 	 .07
0 0 0 	 .12
0, .80 0 	 .72 0 	 .12
0 . 0 0 	 .47
0 0 0 	 .24
.02 .22 0, .07
0, .05 0 	 .04 0 	 .12
.12 0, .40 0, .45
000
.02 .15 .07
0 0 0, .52
0