ANALYSIS OF TECHNICAL GRADE PESTICIDES FOR
N TCDD AT PARTS-PER-BILLION LEVEL
Joseph G. Montalvo, Jr., James F. Ryan, III, and Roy Flagg
Physical and Engineering Sciences Division
Gulf South Research Institute
New Orleans, Louisiana 70136
Number
68-01-3981
rroject uriicer
Patricia Ott
Environmental Fate Branch of the Hazard Evaluation Division
Office of Pesticide Programs
Washington, D.C. 20460
OFFICE OF PESTICIDE PROGRAMS
OFFICE OF TOXIC SUBSTANCES
U.S. ENVIRONMENTAL PROTECTION AGENCY
WASHINGTON, D.C. 20460
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DISCLAIMER
This report has been reviewed by the Environmental Fate Branch of the
Hazard Evaluation Division, U.S. Environmental Protection Agency, and not
approved for publication. Approval does not signify that the contents
necessarily reflect the views and policies of the U.S. Environmental Protec-
tion Agency, nor does mention of trade names or commercial products consti-
tute endorsement or recommendation for use.
ii
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.ABSTRACT
Gulf South Hesearch Institute (GSRI) analyzed 69 technical grade
pesticide batch samples (Silvex; 2,4,5-trichlorophenoxyacetic acid; an
organophosphate; 2,4,5-trichlorophenol; and a chlorinated phenolic antiseptic),
31 replicate samples of these batch samples, and 41 quality control samples
for total tetrachlorodibenzo-p-dioxin (T.CDD). The samples were extracted
with hexane and the extracts were dried, concentrated, and chromatographed
on alumina. Samples were qualitatively and quantitatively analyzed by gas
chromatography/mass spectrometry (GS/MS) in the selective ion monitoring
(SIM) mode. Certainty of TQDD detection was assessed via summation of six
qualitative criteria. Based on this assessment, the data were rated positive,
doubtful, or nondetectable. For those authentic samples in which detection
was judged positive, levels of total TCDD ranged from nondetectable to 30
ppb; quality control samples contained up to 3984 ppb. The use of. high
efficiency capillary column GC/MS is recommended for future TCDD investi-
gations.
iii
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CONTENTS _ _ _
Abstract i iii
Figures v
Tables v±
Acknowledgment - vii
1. Introduction... 1
2. Conclusions 4
3. Recommendations - 5
4. Experimental Procedures 6
Safe Handling in the Laboratory 6
....... General Procedures.."....~:....".". .7."...' .. ...V..."..".. 6
5. Results and Discussion 11
Quality Assurance 11
References . 20
Appendices
A. Data sheets for sample 148919-5-2. 38
B. Master sample status form 41
C. Cumulative tabulated analytical data form 43
D. GC/MS performance ion current chromatograms 45
E. Ion current chromatograms for TCDD detected in a pesticide
sample f. 50
F. Mass spectrum for TCDD detected in a pesticide sample 53
G. Mass abundance tabulation for TCDD detected in a pesticide
sample 55
H. . Ion current chromatograms for nondetectable TCDD in a pesticide
sample 53
I. Ion current chromatograms for questionable TCDD detection in a
pesticide sample 53
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.FIGURES
Number Page
1 Structure of 2,3,7,8-TCDD 2
2 Quality control check for GC/MS performance 12
3 Method blank quality control chart .13
4 Accuracy quality control chart * 14
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TABLES
Number '. Page
21
1 Qualitative Criteria Explanation -
/
2 Dependence of Criterion #6 Overall Binary Characteristic on
Sabcriteria Results. .
3 TCDD Detection Judgement Based on Qualitative Criteria
Characteristics ^
4 "Various EPA Samples' Analyzed" for Total TCDD..... .T. 77. ~. ...".'. .T. ..".""24
5 TCDD Data for Authentic 2,4,5-Trichlorophenol - 25
6 TCDD Data for. Authentic 2,4,5-T (2,4,5-Trichlorophenoxyacetic
Acid).. 26
7 TCDD Data for Authentic Butoxyethyl Ester of 2,4,5-T 27
8 TCDD Data for Authentic Isooctyl Ester of 2,4,5-T 28
r i
1 i ,
9 TCDD Data for Authentic Butoxypropyl Ester of Silvex 29
10 TCDD Data for Authentic Isooctyl Ester of Silvex 30
11 TCDD Data for Authentic Chlorinated Phenolic Antiseptic 31
i
: 12 TCDD Data for Authentic Organophosphate 32
13 TCDD Data for Quality Controls 33
14 Variation of TCDD Detection Judgement Versus EPA Sample Type 34
15 Variation of Total TCDD Level Versus Sample Type for Positive
: Samples i 35
16 Comparison of Replicate Results for EPA Quality Control Samples 36
'17 Comparison of Replicate Results for Authentic Pesticide Batch
Samples ,.
vi
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. ACKNOWLEDGMENT. -
The cooperation of Mr. Ronald ?. Thomas, Supervisory Chemist, Benefits
and Field Studies Division, Environmental Protection Agency, in sampling
design and shipment of the samples to Gulf South Research Institute is
appreciated. The assistance of Ms. Patricia Ott, Project Officer, is
gratefully acknowledged. The assistance of Dr. Mary M. MeKown, GSSI senior
analytical chemist, throughout the project performance is appreciated.
vii
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....SECTION 1 .
TJJTRQDUCTION
Polychlorinated diienzo-p-dioxins (?CDDs) may be formed as undesirable
trace impurities In the production of technical grade chlorophenols . The
latter are employed extensively as pesticides, as wood preservatives and as
starting materials for a series of other products. Through careful control
over conditions in the pesticide manufacturing process, only minimal
quantities of PCDDs are formed, but at temperatures above 200 °C a condensa-
tion of chlorophenols may occur with formation of increasing quantities of
PCDDs (1).
Several PCDDs have been found to be highly toxic, and possibly carcino-
genic and are stable in biological systems (2) . Theoretically, a. total of
75 different PCDDs exist, including 22 tetrachlorodibenzo-p-dioxins . The
toxicological properties of positional isomers of the same dioxin may vary
significantly. Since the 2,3,7,8-tetrachlorodibenzo-p-dioxin (Figure 1) is
extremely toxic (3) , TCDDs deserve special consideration. Although the
synthetic route for TCDD formation during chlorophenol manufacture favors
2,3,7,8-TCDD, other tetrachlorodibenzo-p-dioxin isomers may also be formed
.(*)
Under the aegis of the Federal Insecticide, Fungicide, and Rodenticide
Act (FTFEA), the U.S. Environmental Protection Agency (EPA) has set
TCDD contamination limits in technical grade hexachlorophene at 100 ppb
(0.1 ppm) in order to reduce human and environmental exposure. The purpose
of the GSSI research effort was to determine levels of TCDD present in
technical grade pesticide samples submitted by EPA.
i ' _
In view of the inherent toxicity of TCDD, special precautions in
handling are required during analysis. Concentration by several orders of
magnitude is sometimes necessary, and an ultrasensitive detection method is
preferred. A combination of gas cfaromatography anrf mass spectrometry has
been shown to be an excellent method for detecting TCDDs at very low
levels (5). Identification of individual TCDD isomers requires an initial,
high efficiency separation such as glass capillary gas chromatograpfay (GC)
prior to mass spectrometric (MS) analysis. Low resolution GC columns,
i.e., packed columns, provide a convenient *»»gq for TCDD isomer summation
(total TCDD) by GC/MS.
The present study has focused on determination of total TCDD in
technical grade samples by low resolution gas chromatography/mass spectro-
metry. Sample preparation Included addition of d 2,3,7,8-TCDD to assess
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Figure 1. Structure of 2,3,7,8-TCDD.
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dioxin recovery,, solvent extraction, and cleanup on alumina. GC/MS analyses
were carried out using a. packed column; the standard addition Injection
technique was used.' Selective ion monitoring (SIM) and complete mass
spectral data were obtained. Six qualitative criteria for TCDD detection
were developed incorporating the precision of the analytical measurement
system. The project quality assurance protocol included internal quality
control samples (precision and accuracy) and quality control charting.
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-SECTION 2....
CONCLUSIONS
The results of low resolution gas chromatographic/mass spectrometrlc
analysis of 141 samples (including quality controls and authentic pesticide
samples containing unknown quantities of TCDO) for total tetrachlorodihenzo-
p-dioxin have Indicated, levels ranging from nondetectable to 30 ppb for 69
authentic pesticide batch and 31 replicate samples and nondetectable to
3984 ppb for 41 quality control samples. TCDD was not detected (detection
limit 10 ppb) in 66 percent of the authentic samples, detection was doubtful
in 9 percent and positive in 25 percent. Whereas TCDD could not be detected
in a majority of authentic samples, positive samples contained 10-30 ppb
.total TCDD.
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_.. . SECTION 3 ...
BZCOMMEHDATIONS
The investigation of total tetrachlorodibenzo-p-dioxin levels in
-compounds for whose synthesis 2,4,5-trichlorophenol is used should be
continued. In view of the known toxicity of the 2,3,7,8-TCDD isomer and
incomplete separation of the isomers using packed columns, future investiga-
tions should be performed using high efficiency gas chromatography (capil-
lary column) /mass spectrometry. These data are of major Importance from
the point of view of environmental protection.
In spite of the eminent need for standardized methods for TCDD analysis
in technical grade pesticide formulations and environmental samples, such
procedures are still lacking. The establishment of standardized methods
would permit substantial progress in baseline and ongoing studies. Such
methods should be directed toward improved sample preparation (to reduce
interferences) and improved resolution/detection of the 2,3,7,8-TCDD
isomer, with the consideration in mind that such methods should be suitable
for use by moderately well equipped laboratories.
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.SECTION.4 ..
EXPP.BTMKNTAT, PXQCEDUBES
SAPS HANDLING IH THE LABOBAIOBZ
i
Disposable'rubber gloves vers worn while handling samples and solutions,
and all. evaporations were performed in a hood. Waste materials vere stored
in a capped steel drum labeled TCDD WASTE. Personnel associated with the
project had periodic physical examinations.
GENERAL PBOCEDUBES_
i
After taking custody of samples furnished by the EPA, TCDD residues
vere identified and quantified using a three-stage analytical protocol
including sample preparation* mass spectral data aquisitlon, and data
analysis. ,>.;';:
TCDD Standards
TCDD standards in benzene vere supplied by Dr. Aubry Dupuy, Pesticide
Monitoring Laboratory, EPA. The 2,3,7,8TCDD standard concentration vas
500 picograns Cpg) /microliter C)il) . Concentration of the CL labeled
2,3,7,8-TCDD standard vas 430 pg/ul. Labeled TCDD was used to determine
dioxin recovery through the entire extraction/cleanup procedure. Nonlabelad
TCDD vas used to quantify dioxin levels in the samoles.
Sample Preparation
i
; Ethanol, hexane, benzene, carbon tetrachlorlde, and methylene chloride
vere nanograde (pesticide) quality; inorganic salts, acids and bases vere
ACS grade. Distilled water vas-prepared by filtering through a mixed bed
ion exchanger, activated carbon, and distilling in an ail glass still.
. AirmriTHnq oxide Woelm neutral, activity grade I, vas used for column chroma-
tography. ' :
;. The procedure used In these analyses is a modification of that provided
by EPA in the contract. In general, it involved caustic SDH saponlf ication
of the. pesticide followed by extraction. of, the neutral TCDD into hexane.
The hexane solution vas cleaned up by extracting several times with sodium
hydroxide solution and sulfurle acid, followed by chromatography on aTi-m-ina-
The eluate vas concentrated for GC/MS analysis. Details of the procedures
follow. - - -..-.
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A 1 g quantity of sample was transferred quantitatively Co a. 500 ml
separatary funnel containing 60 ml of pesticide grade etfaanol. Exactly 43
ng of Cl labeled 2,3,7,8-TCDD was added with a microsyringe and the
mixture swirled. Four milliliters of 40 percent KOH was added and the
mixture immediately shaken. The room temperature digestion was allowed to
proceed for 30 minutes, with occasional swirling. Two-hundred milliliters
of distilled water was added to dissolve the precipitate formed upon base
addition, and the resulting solution was extracted with three 100' ml portions
of nanograde hexane. The combined hexane extracts were washed sequentially
with three 100-ml volumes of IN NaOH, one 80-ml volume of distilled water,
and 95 percent H-SO, until no further color appeared in Hj30^' A ^ina^-
washing with 80 ml distilled water completed the extraction.
The fr«»-ggri«» extract was filtered through a 15 mm x 20 en column of
granular anhydrous Na_CO, followed by concentration in a Kuderna-Danish
evaporator to a volume of about 4 ml. The anhydrous Na_CQ_ had been
purified by pre-extraction with nanograde CH-Cl- in a Soxhlet extractor and
drying at 100°C in a vacuum oven. The concentrate was reduced to 0.5 ml
under a stream of UEP or ultra high purity nitrogen (Matheson, Rutherford,
N.J.). The residue was -reconstituted in 2 ' ml" nanograde hexane and chromato
graphed through a 4.5 cm column. The column consisted of activated
over which a 1/4 inch layer of purified granular Na_SO, was placed. The
column was preconditioned by washing with 5 ml of 'CH_C1_, followed by
removal of residual CH-CL, by blowing dry N_ through the column, and heating
for 43 hours at 240°C, Tse sample was eluted from the column with 6 ml
Cd, followed by 4 ml of CH,d-. The carbon tetrachloride layer was discarded
and the C^Cl^ fraction evaporated under UHP N_ to 20 jil. The concentrate
was reconstituted to 2 ml with hexane, evaporated to 20 yl, and rediluted
with hexane to 2 ml. The final extract volume was concentrated to 200 yl.
The three evaporations were necessary to remove residual CH-d_.
f I i
The extract was divided equally into two glass ampules. Ampules were
flame sealed, and the extract volume noted by marking the liquid level.
After coding the ampules were stored in a freezer at 10 °C.
Samples were processed in "sets". Each set of samples consisted of
internal and EPA supplied quality control (QC) and pesticide samples.
Generally each set of samples contained two quality control samples and
four pesticide samples processed from -left to right as indicated below:
-- =.--' .
where A. - ith observation of the accuracy
QC sample (for example, pesticide sample #0093, furnished by
EPA, accepted value for TCDT3 - 2.3 ppm).
X. - the jth EPA pesticide sample (unknown TODD level) ,
.^ \ " ktfa observation of the zero level TCDD QC sample (solvent
blank spiked only with 'd TCDD, 48 ng) .
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QC and EPA sample numbers for each paired extract vere coded with a.
numerical suffix: one ampule, -2 (for immediate GC/MS assay) and the
other, -1 (for backup analysis) . EPA sample numbers ranged from 2 to 9
digits. Replicate samples in this report are denoted by an "I" (initial),
"D" (duplicate), "T" (triplicate), or "Q" (quadruplicate) after the sample
batch number. The series sample number (e.g., 148920) represents one
source of technical grade material; the -1, -2, -3, etc., following the
series sample numbers each represents a different batch sampled from that
source.
TCDD and Cl TCDD standards were transferred to Mininert (Supelco)
valve vials. A back-up seal in the Teflon valve ^d been removed and the
assembly prewashed. Standards were coded and frozen until utilized.
Mass Spectral Data Aquisition
The low resolution gas chromatograph/mass spectrometer used was a
Hewlett-Packard (HP) 5985 GC/MS in the electron ionization (El) source
configuration. The control unit displayed the ion current (peak area) on
a Tektronix 4012 CHT computer terminal ,. and hard copies were provided by ~
the Tektronix 4631 hard copy unit.
Generally spectrometer operating conditions were as follows: column 3
ft x 2 ma IB glass; packing 3 percent 07-11 on 100/120 mesh Supelcoport;
column temperature 200° C for 1 minute to 240*C at 6"C per minute;- injection
heater temperature 250°C; jet separator interface temperature 260°C; ion
source temperature 150*0; carrier gas, helium 32 ml/min; electron impact
source 70 eV; emission 0.3 mA; electron multiplier 2.6 KV; mass range 150-
330 amu; scan <?" 3 ms/amu. The mass spectrometer was tuned to monitor
three molecular ions at m/e values of 320, 322, and 323. Retention time of
TCDD was approximately 3.6 minutes under the above conditions.
The GC/MS protocol for TCDD_measurement was designed to permit simul-
taneous data acquisition for: d TCDD recovery, TCDD detection limit,
TCDD quantification, and confirmation via mass spectra (molecular ion
abundances) . In fM.s procedure, sample extracts and TCDD standards were
thawed to' room temperature. Ampules containing sample extract were broken
above the 1-frp-M level and firmly positioned in a plastic block. TCDD
solutions were always handled under a hood, and liquid and solid wastes
were transferred daily to the TCDD waste drum.
A 10 ul Hamilton 70121 microsyringe was used to inject sample and
standard aliquots into the GC/MS. GC column performance and MS linearity
were checked by co-injecting the following mixtures of standards, one
injection, per mixture in the sequence tabulated:
1 ul native TCDD + 1 ul 37G1 TCDD
. 2. ul native TCDD * 2 ul 37d TCDD
1 ul native TCDD -K 1 ul ^d TCDD
2 ul native: TCDD + 2 ul ^d TCDD
8.
-------
Data for each injection were displayed on the CRT computer terminal.
Peaks (m/e - 320', 322 and 328) corresponding to the TCDD retention time
region were integrated using a HP-21MXE data system and results displayed.
Collected data for each injection were recorded on a Tektronix 4610 hard
copy unit.
Quality control and pesticide samples were run in the same order in
which the extracts were prepared. ...Co-injection of mixtures,, one injection
per mixture, was as follows:
COMPOSITION OF ANALYSIS SET
eo-inj eeted)
Injection Sample Volume TCDD ~" Cl TCDD
Number (ul) Native Standard Standard
1 :
2 -.
3
, 4- _ -....-
2 4
2 4
2 4
^mm _^ ^ O^» ^
0 4
1 4
0 4
.. .. __ .-2 _ 4
0
2
0
-1-
During the TCDD retention time-span the m/e 320, 322 and 328 peak
areas were integrated, and a photocopy prepared for each injection (see
sample hard copies in Appendices). Background noise level was integrated
from the baseline adjacent to the m/e 322 peak. The ion current chromato-
. gram from injection 1 was used for noise determination if the 322 peak was
present; the chromatogram from injection 2 was used if the 322 peak was not
found. A photocopy of .the noise level was prepared.
I
Hard copies from injections 1 and. 3 were inspected to determine if
both m/e 320 and 322 peaks were found for both injections. A sample
meeting these criteria was subjected to GC/MS mass fragmentation confirma-
tion by evaporating the extract under N_ to about 10 ul. Two microliters
of the concentrated extract was injected. Both mass spectrum and absolute
abundances for m/e values were obtained and hard copies prepared (see
sample copies in Appendices).
1 I
Data Analysis " - :. .1- :
Integrated peak areas for TCDD standard solutions were subjected to
linear regression analysis (y " peak area; 2 ul injected) . Correlation
coefficient (r), slope (s), intercept (i), and the ratio i/s were computed
for m/e - 320, 322, and 328. Instrumental sensitivity, expressed as area/pg,
- - -J fof m/<" "
cc
concentration in pg/ul) and 480 C*'ci TCDD standard concentration in pg/yl) .
An average 320/322 ratio was computed from the (2 + 0 + 0) injections
Pertinent data were plotted on a QC chart to monitor instrument performance.
Expressed as percent, i/s s 100, average values were < + 5 percent. Large
negative values, i.e., 20 percent, could be indicative of TCDD adsorption
or degradation in the system. In any case, this problem was usually reeti-
.-f.*ed bcging theGCcolumn. ~
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Peak areas for the method blank (QC), accuracy (QC) , and EPA samples
were tabulated on data sheets, designed exclusively for this project, along
with other pertinent information. A-sample data sheet is shown In Appendix
A; the method for computing percent Cl TCDD recovery, TCDD sample concen-
tration, detection limit, and results of the confirmation analysis are also
illustrated. Note that reported TCDD levels are corrected for recovery.
The contract specification for Cl TCDD-recovery in the pesticide
samples was 50 to 120 percent, inclusive. If the initial GC/MS recovery
assay was outside the specified range, the backup aliquot of the original
extract was assayed- If the average of the two recovery values was within
the specification range, contract requirements were satisified. If the
average recovery was outside the specification rang«,. the sample was re-
extracted and assayed one additional time. Confirmation of TOD detection
by recording complete mass spectra was optional at TCDD contamination
levels <10 ppfa and a contract requirement at XLO ppb.
The standard addition technique was not successful with samples con-
taining large quantities (parts-per-nillion) of TCDD because the amount of
TCDD standard co-injected was Insignificant compared to the TCDD level in
the pesticide extract. Determination of regression slope and intercept for
TCDD quantifications with such samples was variable due to poor correla-
tions. An alternative GC/MS protocol was used for these, samples. An
external calibration curve was constructed by injecting varying amounts of
the TCDD standard into the GC/MS. The resultant linear regression equation
for- the standard, in terms of picograms rather than microliters injected
was: ; . .., '
y - X: s V i - (1)
. j
where y - peak, area (m/e » 322)
s » slope (area/pg TCDD)
i - intercept
X m pg ICDD Injected
Rearranging Eq., (1):
I
Xp - (yp-i)/s (2)
'where £ * pg TCDD in the pesticide sample extract and y =» average m/e 322
area fur a 2 jil. sample extract. TCDD concentration (ppm? in the sample is
i equal to: ' '
' - »- , ; -,-^:..' ~' ... ".' - ,.."-- i ,.-- . - .
" ~ t >
_ (200 ul sample extract volume) , - __,.
W« Xo ,.8-tt ul inlected into GC/MS) /CR'? saffiple
: .. .^- . -.-*.- ^j
where S is the percentage recovery of Cl TCDD.
10
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... SECTION 5 ..
RESULTS AND DISCUSSION
QUALITY ASSURANCE
Several aspects of this project vere monitored for quality assurance.
These included (1) taking custody of standards and samples, (2) materials
and reagents used in the analysis, (3) standard procedures, (4) calibrations,
(5) record keeping, and (6) quality control charts. Chain of custody
records were transmitted to GSRI along with samples and standards. Sample
receipt was acknowledged by dating, signing, and returning a copy of the
chain of transfer record to EPA. Discrepancies in labeling or replacement^ .
requests were noted on the transfer record.
Materials and reagents used in the analysis in .general were ACS or
pesticide grade. New lots of material were tested for TCTD interference by
subjecting them to the analytical protocol. The sample extraction and
clean-up procedure described in Section 4 was used throughout the project,
without modification, and therefore constituted a standard operating proce-
dure. Generally, the GC/MS protocol followed the standard procedure outlined
in the previous section; certain modifications, i.e., changes in column
temperature programming, were necessary to overcome interferences and
refine the method. :
. '
The GC/MS instalment was tuned daily prior to analyses. GC/MS perform-
ance and linearity standards bracketed 56 successive analyses. Record
keeping Included an instrumental preventive maintenance log. GC/MS data
and computer file reference number, date of extraction, «"«* the sample
weight were recorded in .this log for each sample assayed.
Sample processing accountability was an important part of record
keeping. Results were compiled on a master sample status form (see Appendix
B) in the order samples were received. Results were also compiled on a
cumulative data: form (Appendix C) in the order samples were assayed.
Cumulative quality control charts were useful indicators of whether or not
the measurement process was in control. The charts showed trends or runs,
sudden shifts in the mean, increased variability a?M often indicated the
nature of a problem.
i i-
i
Figures 2 through 4 show quality control data for GC/MS performance,
method blank,, and. accuracy of TCDD determinations. Control chart analysis
of graphs A, B, C, and D (7igure 2) shows systematic variations with
probable causes. A series of increasing and decreasing values (graphs B
and D, observation numbers 1-6 and 29-34) was obtained after the instrument-
_n
-------
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1 1 1 1 1 1 1 1 1 1
0 5 10 15 20 25 30 35 40 4!
OBSERVATION NUMBER
& 37 35
Figure 2, Quality control chart for GO/MS performance (Sample: Cl TCDD and Cl TCDD standards) . i
-------
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. . AM Observations
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1 1 1 1 1 1 1 1 i
0 6 10 15 20 25 30 35 40 45
. OBSERVATION NUMBER
Figure 3. Method blank quality control chart (Sample: solvent blank with added Cl TCDD) . t
i
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-------
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OBSERVATION NUMBER
Figure 4. Accuracy quality control chart (Sample t EPA
All Observations
n-19
3? « 96.7%
1 >_! Jfc f\t
8 **37.8 ^
CV39.|%
n-17
XB2.25ppm
5 B0.38ppm
CV»|7%
1 ' 1
35 40 4
00093) .
-------
was shut down and the ion source cleaned. Observation numbers 18- ^
35-39 show decreased sensitivity prior to source maintenance. "V
cfaromatograms for observation 21 are shown in Appendix D. (j^a°
lower sensitivity, runs of 2 to 4 observations (graphs A and C) were
with regression correlation coefficients <0.980. Nonetheless, grapn *
shows that only 1 observation out of 41 was outside the arbitrarily cnos
+3 standard deviation limits for 320/322 ion current peak areas, xne ^
theoretical ratio of 0.766 (6) compares favorably with the observed mean
value (X - 0.784, percent difference - + 2.3 percent). The conclusion was
that the measurement process (320/322 ratio) was in control for 97.6 percent
of the observations is based on Duncan's (7) criteria that define statis-
tical control to be any point within 3 standard deviation limits. Ho ;
Mm-ffg were placed on AD graphs.
: Figures 3 and 4 show the method blank (solvent spiked only with Cl
labeled TCDD) and accuracy (EPA reference pesticide sample #0093) quality
control charts 'with EPA recovery specification range. Mean recovery observa-
i tions falling within SPA specifications were 81 and 86 percent, respectively.
The EPA accepted value for total TCDD concentration in the pesticide
reference sample is 2.3 ppm (8). A mean of 2.25 + 0.38 ppm was obtained,
with a coefficient of variation of 17 percent. No TCDD was observed la any .
method blank.
; Several recovery observations were outside the specification range.
A run of out-of-specification values (Figure 3, curve A, observations 68)
was due in part to a partially clogged jet separator. Cleaning the jet
separator rectified the problem. Random observations outside specification.
limits probably reflect overall variance of the total procedure.
j " A majority of out-of-specification recovery values were on the high
side, i.e. >120 percent, probably because the upper onesided tolerance of
the analytical process is only 20 percent above the theoretical recovery
value of 100 percent; the corresponding lower one-sided tolerance is 2.5
times as great, i.e., 50 percent.
; Criteria for a Positive Sample
- Qualitative criteria developed for the detection of TCDD in the
pesticide extract matrix were: (1) a recovery of d TCDD between 50 and
120 percent, inclusive; (2) both m/e 320 and 322 peaks existing at the TCDD
retention time region; (3) well defined m/e 320 and 322 peaks whose retention
tine exactly equals that of TCDD standards; (4) signal-to-noise ratio of at
least 2.5/1 at m/e 322; (5)-ratio of m/e 320/322 peak areas in the proper
isotopic proportion; and (6) TCDD confirmation by molecular ion spectra.
Data for the first five requirements were obtained by selective ion monitor-
ing at m/e 328, 322, and 320; data for TCDD confirmation were generated by
recording spectra in the m/e range 150-350 and computer tabulation of
absolute abundances of 119-131 m/e. The range of interest was m/e 320-323;
the wide range facilitated background measurement. Pertinent information
explaining these binary (positive/negative) decision making processes is
shown in Table 1. . --
15
-------
Criterion $1 was based solely on recovery specifications in the
contract. Criteria 5 and 6 incorporate variance associated with the
measurement system. Statistical, limits imposed are shown in Table 1. The
theoretical isotopic abundance ratio of m/e 320/322/324/326 relative to 322
is 0.766/1.000/0.489/0.001 (6). Criterion #6 was divided into three
subcriteria (Table 1); eight binary permutations of the subcriteria are
possible as shown in Table 2. Overall criterion #6 test characteristic was '";
arbitrarily chosen from the three subcriteria.' The rational for" subcriteria
decision making correlates with isotopic abundance ratios. Greater emphasis
was given to 6a (m/e 320/322) since these tvo m/e values are the largest.
More precise ratio measurements are anticipated with 6a rather than the
smaller 324/322 ratio for 6b or 6c. Table 3 shows the evaluation of the
certainty of TCDD detection based on summation of qualitative criteria
characteristics. Criteria 1 through 4 were absolute or necessary for
determining if TCDD was detected except for the D (duplicate) , T (triplicate)
and Q (quadruplicate) reruns of original samples. Detection limits and
mass confirmation spectra were not required for the latter.
Appendices DI include representative hard copy data for GC/MS instru-
mental performance and TCDD detection. "'-
!
Sample Data
A wide variety of pesticide samples were analyzed for total TCDD
residues. Submitted EPA samples (unknowns and quality control samples)
were analyzed on a blind basis. TV two sample types were visually and
experimentally indistinguishable. EPA provided decoding information only
'after completion of the. assays and reporting of the TCDD data. Table 4
: summarizes the decoding Information. Generally, a 6-digit sample code
;represents a series of pesticide samples of the same generic class; indivi-
dual yr"pl.«« were denoted by a hyphenated (in some cases) suffix made up of
numbers and/or letters. For example, 127332-5D relates to the duplicate
determination of gfre fifth subsample in the chlorinated phenolic antiseptic
137832 series. Each subsample represents a different batch of the manufac
; Cored technical grade pesticides.
i t- *
' Based on EPA decoding information, replicate sample data, if available,
were collated according: to sample batch number. After first combining
; replicate information with the appropriate first-run results, a TCDD detec
: tion judgement was made for each, batch sample, regardless of the number of
' replicates assayed. These judgements were either positive, negative, or
; doubtful. For example, if the first run (I) sample of batch number 1234
' was positive (e.g., 12341 is +), the duplicate (D) run negative (1234D is -),
' and the triplicate- (T) run questionable (1234T is doubtful), then the
judgement was that TCDD detection was doubtful in sample 1234. On the
' other hand, if. a sample batch were judged TCDD positive, regardless of how
. many replicates were run, total TCDD level found was reported, including
:-replicate data and the average value, if appropriate.
':' A total of 69 authentic pesticide batch samples were assayed. TCDD
.was detected in 17 batch samples (75 percent) ; TCDD detection was doubtful
in 6 samples (9 percent) ; and- TCDD was not detected in 46 batch samples (66
1.16'.
-------
percent). Tables 5-12 summarize data for TCDD la various types of
pesticide samples. In those instances where an or most batch samples were
positive for a given series, tabulation of mean value, standard deviation,
and coefficient of variation is based on positive sample data. NA tnot
applicable) refers to information not available or not requested. For
example, qualitative detection criteria #4 and 06 were not required for -
D, -T, or -Q samples. Data for EPA quality controls are shown in laoie ^
13. - - " "
Variation of TCDD detection judgements versus EPA sample type is shown
in Table 14. For the authentic batch samples TCDD was nondetectable
(detection limit 10 ppb) in 100 percent of the 2,4,5-trichlorophenol
samples analyzed (n, number of batch samples assayed - 16). For compara-
tive purposes, batch samples of 2,4,5-T, the butoxyethyl ester of 2,4,5-T,
and the isooctyl ester of 2,4,5-T are grouped under 2,4,5-T as shown
below.
Type
Number
of Batches
of Batches /(percent)
Positive Doubtful Nondetectable
2,4,5-T
butozyethyl ester
of 2,4,5-T
isooctyl ester
of 2,4,5-T
l/(33.3) l/(33.3) l/(33.3)
10
_3
16
9/(90)
_0
10/(63)
I/(10)
I/(100)
£
1(6)
Overall, detection of TCDD was positive in 63 percent (n»10) of the batch
samples, doubtful in 31 percent (n-5), and nondetectafale in 6 percent
(n-1). Specifically, detection of total TCDD was" positive in 90 percent
(n-9) of the butoxyethyl ester of 2r4,5-T batch samples. In contrast,
detection of TCDD was judged equally positive, doubtful and nondetectable
in all (n-3) of the 2,4,5-T batch samples, and TCDD detection was questionable
in:'all (n-3) of ti,e isooctyl ester of 2,4,5-T batch samples.
Eight Silvex type batch samples were assayed. TCDD detection was
positive in 86 percent (n-6) of the butoxypropyl ester of Silvex and question-
able in the remaining sample (n-1) of this type. Only one batch of isooctyl
ester of Sllvex was assayed; the TCDD detection judgement was positive.
.One-hundred percent of both the chlorinated phenolic antiseptic (n-22) and
the organophosphate (n-7) batch samples gave negative TCDD detection charac-
teristics. - -
17
-------
Summarizing the TCDD detection judgements for authentic pesticide
batch samples In terms of decreasing percentages, 66 percent were nondetect-
able, 25 percent positive, and 9 percent doubtful. Likewise, judgements
for the 20 different EPA QC samples analyzed were 40 percent oondetectable,
30 percent doubtful, and 30 percent positive.
Variation of total TCDD level found versus sample type for positive
samples is shown In Table 15. All authentic pesticide batch samples for'
which TCDD detection was judged positive contained total TCDD In the range
10-30 ppb. By contrast, total TCDD content of EPA quality controls varied
from a low range of 31-99 ppb to a high of 3000-3499 ppfa.
Precision of TCDD determinations in the EPA quality controls is shown
'in Table 16. Relative standard deviation ranged from 2.6 percent to 33
percent. Standard deviation range- for authentic unknown replications was
.3.1 percent to 52 percent, as shown in Table 17.
The prevalence of doubtful TCDD detection judgements in authentic
pesticide samples and EPA.' quality controls suggests the need for Improvements
in the measurement system. A significant number of doubtful detection
instances are related to apparent TCDD levels <10 ppb; in addition, TCDD
confirmation was not a contract requirement at these low levels. Mean
detection limit was 4.7 ppb ±3.5 ppb (standard deviation). Mean value
plus two standard deviations =-11.7 ppb; thus questionable TCDD detection
at the 10 ppb level can be expected. Therefore, any result found below 10
ppb in authentic samples was reported as nondetectable (ND). Actual values
below 10 ppb in the. quality controls are tabulated in Table 13 to demonstrate
the Inaccuracy in reporting such data. The qualitative criteria were used
to define the judgement below 10 ppb (i.e., positive, doubtful, or negative).
Problems encountered in tha project - low- and high recoveries, unrelia-
bility of standards, inability to distinguish matrix problems from instrumen-
tal ones, variability in machine sensitivity, changes in chromatography,
and interferences - contributed to observable variations in relative standard
deviation and detection limits. Extracts yielded interferences of varying
severity from other, substances depending upon the efficiency and r-eproduci-
bility of the cleanup procedure and sample type. Varying interferences
also affected Cl TCDD recovery. For example, excellent recoveries were
obtained for samples- 148921-3, 148921-3D, and 143921-4D. Zero percent
recoveries were obtained with 143921-3T and 143921-4T. Internal quality
control samples in the same sample sets (pertaining to extraction) gave
excellent recoveries. .--...., i ....
Low recoveries resulting, from Interferences might be prevented by
employing a second Al_0_ cleanup column or other modification in the
extraction procedure. Recoveries higher than the theoretical value were
anticipated when We 323 peaks, co-eluted with CL-TCDD.
Interfering m/e 320 and 322. peaks co-eluting with TCDD contributed to
variability in calculated TCDD levels. In some instances peaks elating
prior to or immediately after TCDD prevented accurate peak interpretation.
-------
Application of high resolution gas stenography should help to Wrcame
problems with interferences. ,
Problems with standard reliability included irreproducibility of lots
and concentration upon standing. TCDD standards were provided by EPA and
lots were checked against each other to identify suspect solutions. TOD
standards were provided in benzene solvent. Aliquots of the standards were
transferred to small vials sealed with Teflon-lined rubber septums. _ - -
Standards were withdrawn with a 10 jil micro syringe. Repeated insertions of
the microsyringe needle (four insertions were required per sample assay)
resulted in formation of holes in the septum. Benzene evaporated through
the porous septum and concentration of the TCDD standard increased. An
attempt was made to solve the problem by adding a small amount of mercury
to the standard vials to seal punctures in the septum. Between Inj ections,
vials were stored in an inverted position which prevented benzene evaporation.
However, after a few days the solution turned cloudly, indicating instability
of the standard solutions. The problem was rectified by using Mininert
vials as outlined in the experimental section.
Variability in machine sensitivity contributed to variable .TCDD
detection limits and regression correlation coefficients. Approximately
one hour was required to complete the four GC/MS runs needed to construct
the regression line. Drift in GC/MS sensitivity and specificity over this
period contributed to a poorer correlation coefficient with increased
variability in reported TCDD recovery and quantification.
Distinguishing matrix problems from instrumental ones was difficult.
Replicate recovery assays of the same extract provided data to elucidate
problems. Poor precision was indicative of instrumental problems. High
precision combined with recoveries outside the specification range suggested
matrix or extraction problems. :
Total TCDD was judged nondetectable in over 50 percent of the authentic
pesticide batch samples assayed. Extending the lower workin,g range of the
measurement system seems desirable. TMs task might be accomplished by
further cleanup of the extract, evaporation of the. extract to a smaller
volume, and employing high efficiency gas cfaromatography/mass spectrometry.
! ----- .--': - - -:,.-.- - - ...
Application of the more tedious standard addition technique to these
"analyses appears justified over external calibration. Variations in the
linear regression slope were observed for some samples, which may be indica-
tive of matrix effects. The standard addition technique also tends to
compensate for instrument drift. Therefore, more accurate .data would be
anticipated with the standard addition approach than with external working
curve techniques. ' ..... -
19
-------
REFERENCES
1. Blair, E.H. Chlorodoadns-Origin and Fate. Preface, Advances in
Chemistry Series #120, American Chemical Society, Washington, D.C.,
1973. p. 141.
2. Scnwetz, B.A., J.M. Norris, G.L. Sparschu, V.K. Rowe, P.J. Gehring,
J.L. Emerson, and C.G. Gerbig. Toxicology of Chlorinated Dibenzo
ParaDioadns. Environ. Health Perspect. 5:87-99, 1973.
.3. IARC (WHO). Some Fumigants, the Herbicides 2,4-D and 2,4,5-T, Chlorin-
" ated TKfr<****ni]. 1qytTtg and Miscellaneous Industrial Chemicals. ~" IARC
Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to
; Man, 15, 1977. 354 pp.
4. Rappe, C., H.K. Buser, and H.P. Bosshardt. Identification and Quantifi-
cation of Poly-Chlorinatad Dibenzo-p-Dioxins (PCDDs) and Dibenzo-
Ferans (PCOTs) in 2,4,5-T-Ester Formulations and Herbicide Orange.
Chemosphere, 7(5): 431-438, 1978.
,5. Firestone, D. The 2,3,7,3-Tetracfalorodihenzo-p-Dioxin Problem: A
:. Review. In: Ecological Bulletins (Stockholm), 27:39-52, 1973.
r * f
6. Beynon, J.H. Mass Speetrometry and its Applications to Organic Chemis-
try. D. Van Nostrand Co., Princeton, M.J. 1960. 640 pp.
7. Duncan, A.J. Quality Control and Industrial Statistics, 4th ed.,
Richard D. Irwln, Inc., Homewood, 111., 1974. p. 207.
r
.3. Personal Communication, Mr. Ronald Thomas,. Environmental Protection
Agency, Beltsville, Md., 1978..
:-20
-------
LBLE 1. QUALITATIVE CRITERIA EXPLANATION,
Ho.
Criteria
Explanation
502) m/e 324/322 control limits set by
mean value + 3 standard deviations as
determined by quality control data
obtained with EPA reference pesticide
sample #0093.
(c) Existence of m/e 320, 322, 324, and
326 in, absolute abundance table.
21
-------
TABLE 2. DEPENDENCE OF CSTTE2ION 06 OVE2ALL BINAEI CHABACTERISTIC ON
STJBCSITERIA RESULTS :
Overall Pennutaclou
Criterion i?6 Characteristic 6a 6b 6c
(negative)
(positive)
22.
-------
TABLE 3. TCDD DETECTION JUDGEMENT BASED ON
QUALITATIVE CRITERIA CHARACTERISTICS
ualitative Criteria No.
Judgement
For First-Run Samples:
TCDD detected
4- _
Any or
negative
NA
TCDD detection
TCDD not detected
For Duplicate (-D), Triplicate (-T), or Quadruplicate (-OJ Samples:
NA
NA
NA
NA
Any or all
negative (-}
TCDD detected if also
detected in first-run
sample
TCDD detection question-
able if first-run
sample also questionable
or not detected.
TCDD detection question-
able regardless of
firstrun judgement
TCDD not detected
regardless of first-
run judgement
aQualitative criteria 1 through 4 all absolute or necessary
for TCDD detected (positive) judgement.
m not applicable.
23
-------
TABLE 4. VARIOUS EPA SAMPLES ANALYZED FOR TOTAL TCDD
Sample Series
Number of
Batch Samples
Pesticide Sample Type
(Generic Name)
137401
137402
137403
137832
137834
137833
137835
148917
148918
141996
141998
141997
148919
148920
148921
229617
226617
225511
222341
: 222455
220777
' 93-01
14-01
0093
1-01
305
304
; 303
90
: ' -.,. UL .
: " ' " 4 ' ~ '
3r^y
n
3 X
3 T
32''.
1
. x
1
14
8
2
. - 4
5
5
- . 1
Z
- ... . '._ ... L-. -
10
7
7
AUTHE^mC PESTICIDES
2,4,5-T*
chlorinaced phenolic
antiseptic
2,4,5-trichlorophenol
isooctyl ester of 2,4,5-T
- isooctyl ester of Silvex
butc33cyethyl ester of
2,4,5-T
butoxypropyl ester of Silvex
organophosphate inseceicide
QDALITJ CONTROLS
i
a2,4,5-T is 2,4,5-crichlorophenoxyaceCic acid.
is 2,4,5-T2 .. - -.-. --
24
-------
* t
EPA
Series
Number
EPA
Batch
Sample
Number
TABLE 5'.
Percent
Recovery
C137TCDD
TCDD DATA FOR AUTHENTIC 2,4,5-TRICHLOROPHENOL
(16 Batch Samples) ; I
Section Qualitative Criteria
(ppb) 1 2 3 A 5 6
Judgement
FOB. Dtfl.a ND
Total
TCDD
(ppb)
137833
137835
148917
148918
8Dtfl.
-1
~2
, -1 -V!"
. ' -2 -:-;1--
.-'-'. -3
".': -4 ,/V
t. .. .' "
" ' ' -2'-,l\
.'; . a .-,- '
-4
-5 '''-''-i;
-1 .:".,.'.
-2 ,".
'-3 ' " ''!
-4 ' ]JrCJ*
"5 ' \
- Doubtful !
95
79
102
::; : 104
82
89
s ;, '
89
69
- . 100
80
86
" , ? t -
51
. 93
96
95
.1
4
3
3
5
9
2
2
1
2
^
8
2
1
5
2
5
1
.6
.5
.7
.0
.3
.1
.6
.9
.5
* J
.8
.5
.8
.6
.6
.0
.7
t- - NA
-f - NA
+ - NA
+ - NA
+ - NA
+ - NA
. + - NA
+ + -f
+ - NA
i»rv
f - NA
f - NA
+ + +
-t- - NA
+ - NA
+ - NA
f - NA
NA
NA
NA
NA
NA
NA
NA
4-
NA
im
NA
NA
+
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
4-
NA
NA
+
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA .
NA
NA
NA
NA
NA
NA
NA
NA
NA
/ ND
/ ND
1 / ND ;
! / ND ,
' / ND
/ ND
/ ND
! / ND
/wn
nif
/ ND
/ ND
1 / ND
i / ND
. / ND
/ ND
/ ND
1
-------
TABLE 6. TCDD DATA FOR AUTHENTIC 2,4,5-T (2,4,5-TRICHLOROPHENOXYACETIC ACID)
EPA
EPA Batch
. Serieq Sample
. Number Number
1 137400 -137401
' -137402
-1374021
1 -137402D
-137403
-1374031
-137403D
Percent
Recovery
C137TCDD
71
68
105
71
58
Detection
(ppb)
2.4
5.2
NA
4.2
NA
Qualitative Crlterl
12345
4- 4- 4- 4- +
4- f 4- 4- -
4- 4- 4- NA 4-
+ 4-4-4-4-
4- - NA NA NA
Average
Total Total
a Judgement TCDD TCDD
6 , Pos. Dtfl.
NA
4- /
NA /
i /
4- /
NA
ND (ppb) (ppb)
/ ND
18
; 17
18
' Dtfl.
i 14
/ ND
Probably false positive since the duplicate sample (137403D) was ND.
-------
TABLE 7. TCDD DATA FOR AUTHENTIC BUTOXYETHYL ESTER OP 2,4,5-T
(10 Batch Samples)
EPA
EPA Batch
Series Sample
Number Number
148919 -1
-«
-ID
-2
-21
-2D
-3
-4
-.-5 .'
i -51 i. '
-5D
-6
-6D
-6T
-7
-71
-7D
-8
-81
-8D
-9 ,;
-91
-9D
-9T
-10
-101
-10D
Percent
Recovery
C137TCDD
80
114
67
99
104
82
89 :>;,>:
114
inn
LWO
78
72
53
111
117
116
97
70
99
1. 1
61
104 i
Statistical information for
Detection
(ppb)
6.8
NA
2.5
NA
8.0
6.4
4.3
NA
» A
.J. V
NA
1.1
2.0
NA
1.8
NA
3.1
NA
4.5
3.5
NA
TCDD positive
Qualitative Criteria Judgment
12 3 4 5 6 Pos. Dtfl. NO
+ 4-4-4-4-4- /
+ 4- 4- NA 4- NA /
/
4- + + + 4- 4- /
4- 4- 4- NA 4- NA /
+ 4- 4- 4- 4- 4- /
4- 4- + + + + /
/
4- 4- 4- 4- 4- 4- /
+ 4- + NA 4- NA /
4- 4- + NA + NA /
4- 4- + 4- 4- + /
/
4- + 4- + 4- /
+ 4- 4- NA 4- NA /
/
+ + 4- + + 4- /
4- 4- 4- NA + NA /
/
+ + + + 4- + /
+ + 4- NA 4- NA /
+ 4- 4- 4- + 4- /
/
+ + 4- 4- + 4- /
+ + 4- NA 4- NA /
batch samples! n
mean total TCDD (ppb)
standard deviation (ppb)
coefficient of variation (X)
Average
Total Total
TCDD TCDD
(ppb) (ppb)
21
23
19
23
25
21
24
29
23
22
23
25
22
21
Dtfl.
33
19
22
14
30
22
23
23
19 .
23
27
19
7
22.7
1.3
5.7
-------
TABLE 8. TCDD DATA FOR AUTHENTIC ISOOCTYL ESTER OF 2,4,5-T
CO
EPA '
Series
Number
141996
141998
. EPA
Batch
Sample
Number
-1419961
-141996D
-141996T
-141996Q
-1
-11
-ID
-2
-21
-2D
Percent '
Recovery
C137TCDD
103
61
78
103
110
73
108
79
Y « £
nlTI n Qualitative Criteria Judgemen
Detection
(ppb) .123456 Poa. Dtfl.
!
2.0 + + * + -* /
NA + + + NA * NA /
5.9 + + + + + + /*
NA * f + NA - NA /
\ *
2.5 + + + + + + /
NA + + + NA - NA /
^
2.2 + - NA NA NA NA
1.5 + - + + + + /*
Total
t TCDD
ND (ppb)
15
30
. 21
27
28
13
/ ND
27
Average
Total
TCDD
(ppb)
Dtfl.
Dtfl.
Dtfl.
Other replicate(a) doubtful or nondetectable, therefore, probably not dloxin-contaminant or inter-
ferencea present,
-------
TABLE 9. TCDD DATA FOR AUTHENTIC BUTOXYPROPYL ESTER OF SXLVEX
EPA
EPA Batch
Series Sample
Number , Number
148920
1
-2
-21 ;'
-2D '-;'
-2T :.,
-".'. -3 ;
-..; -31 -;'
. -3D ;.;
. .-4 .'"
-41 '
-4D ''
' .'?.' -5 .:.
-51
-5D
-5T
-6
-7
-71 '
-7D
I
Percent /-
Recovery D
C137TCDD
'? ' 71
'.'. _ .
; 83
:>;' 115
,,,!% 83. ::
,
'-#.:'/ 66' - "
:;.'f;;-77 - -
v;^.- 67 " '
.a -..',;ii3 . .'.'
-/- ' 97
'^'- \"- 79 ' '
66
103
I ,iri i74 i- M
111
i t ' of '
flTa^tian Qualitative Criteria
0f.6CC.LOn
(ppb) 1
4.4 4-
2.3 4-
NA 4-
6.3 +
5.5 4-
NA 4-
1.0 4-
NA 4-
5.2 4-
NA 4-
7.2 -f
0.9 4 4-
IP
5.6 4-
NA 4-
2345
4- 4- 4- +
4- 4- 4- +
4 4- NA +
4- 4- 4- +
4- 4- 4- 4-
4- 4- NA 4-
4- 4- 4-
4- 4- NA 4-
+ 4-4-4-
4- 4- NA 4-
4- 4- 4- -
4- 4- 4- +
4- + + 4-
4- 4- NA 4-
6
4-
4- '
NA
4-
4-
NA
NA
NA
1
4-
NA
4-
4-
+
NA
Average
Total Total
.Judgement TCDD TCDD
Pos. Dtfl. ND (ppb) (ppb)
/ ' '21
/ 19
/ 19
/ 17
/ 22
/ 19
/ 21
/ 17
/ Dtfl.
/ 20
/ 20
/ 21
/ 24
/ 17
/ 23
/ 12
/ 22
/ 22
/ I 21
Statistical Information for TCDD positive batch samplest n
4
Mean total TCDD (ppb) 20.3
Standard deviation (ppb) 1.5
Coefficient of variation (X) 7.4
-------
TABLE 10. TCDD DATA FOR AUTHENTIC ISOOCTYL ESTER OF SILVEX
EPA
EPA Batch Percent Limit of
Seriea Sample Recovery Detection
Number Number C137TCDD (ppb)
141997 141997 81 9.7
Total
Qualitative Criteria Judccmtnt TCDD
123456 , Pos. Dtfl, ND (ppb)
+ + + + + + ^ 20
Ul
Q
-------
TABLE 11. TCDD DATA FOR AUTHENTIC CHLORINATED PHENOLIC ANTISEPTIC
(22 Batch Samples)
EPA
EPA Batch
Serlea Sample
Number Number
137832
-1
-2
-3
-31
-3D
-4
... -41 . ;,,.-,,
-4D
-5
-51
-5D -
-6
-7
-8
-9
-10
-11
-12
-13
-14
137834 '
-1
-2
-3
_4 i
-5'
-6
-7
-8
Percent
Recovery
C137TCDD
._
83
77
102
68
i 50
61
91
79
90
84
93
68
64
78
74
96
68
93
111
91
78
100 ' '
93
117
102
Limit of
Detectloi
(ppb)
16
12
8.9
NA
1.8
NA
4.0
NA
3.9
9.4
2.2
8.4
3.6
2.3
4.4
2.7
2.6
7.1
2.4
1.9
3.0
1.7
2.3
4.3
1.9
Qualitative Criteria . ,
1
1 2
4-
4-
4-
4-
4- ~
4- -
4-
4. _
4-
-4- _
4-
-4- .
+
4-
4- »
j, _
4-
4-
-4-
4- -
4-
4-
4* *-
4- -
4- -
3
NA
NA
NA
NA
_
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
.NA
NA
NA
4 '
NA
NA
NA
NA,
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
5
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
uuugcui^»««
6 POB. Dtfl. ND
NA ! /
NA j /
1 /
NA ' /
NA i /
/
NA J
NA /
/
NA /
NA /
NA /
NA /
NA /
NA /
NA /
NA /
NA /
NA : /
NA /
NA /
NA 1 /
NA | /
NA i J
NA j /
NA . ; /
NA j /
NA J , /
Total
TCDD
(ppb)
ND
ND
ND
ND
.
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
' ND
ND
ND
ND
ND
-------
TABLE 12. TCDD DATA FOR AUTHENTIC ORGANOPIIOSPHATE
EPA
Series
Number
148921
EPA
Batch
Sample
Number
-1
-2
-3
-31
-3D
-3T
-4
-41
-4D»
4T . ' T' '
-5
-6
-7
. *
Percent Limit of
Recovery Detection
C137TCDD
51
110
100
100
0
111
95
o
78
69
93
(ppb)
9.9
2.9
5.2
NA
NA
2.2
NA
NA
1.5
4.5
9.3
f- TT-.:f -_-_-! -! t " -
Qualitative Criteria . ..., 1
1 23
4- - NA
4- - NA
4- - NA
t - NA
- - NA
4- - NA
4-4-4-
- - NA
4- - NA
4- - NA
4- - NA
4
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
,
5
i
NA
NA
NA
NA
NA
;
NA
4-
NA
NA
NA
NA
4J UU H CUIB tl *-
6 Poa. Dtfl. ND
NA /
NA ' /
1 /
NA /
NA /
NA /
/
NA , | , /
NA * .
NA . /
NA .' /
NA ! /
NA - /
Total
TCDD
(ppb)
ND
ND
ND
ND
ND
ND
ND
u
Probably false positive due to contamination or interferences present, since replicate samples were ND.
-------
13. TOO DATA FOR QUAIHT COSTROLS
l^HBMMMM^
EPA
QC
Humocr
222455
2224551
222455D
220777
2207771
2207770
226617
2266171
2266170
2255U
22351U
2235110
229617
229617T
229617D
1-01
1-011
1-010
1-01T
11
111
110
11T
3H
3T
3TI
3TO
3Z
3ZZ
3xr
4
41
40
14-01
14-011
14-017
3Z
3ZZ
32)
222341
2223411
2228410
0093
90
901
90D
303
3031
303D
304
3041
304B
303
3031
3030
"Sis:
MM^MHMMMMIMB
C1J7TCDD
92
76
71
119
95
116
76
77
58
82
101
61
94
SO
34
100
77
59
S3
83
92
89
115
113
90
83
107
39
68
106
89
76
97
38
60
79
U
98
72
116
94
* doubtful or
M^M.^HM^H^«^..^
Unrlt of
(P?b)
3.1
HA
- 2.3
KA
4.9
KA
5.9
KA
3.6
KA
KA
0.7
HA
HA
9.9
4.9
KA
2.4
KA '
HA
5.5
13
KA
3.1
HA
HA
2.5
KA
2.0
KA
2-*V
'. 'l.2" . :
HA
KD at oehar re
i i. . m»i» " i "
Oiutlitacivc Criteria
1234
* - KA HA
+ + + HA
4 * + KA
4- + 4- HA
f - SA MA
+ 4 + KA
+ * -t- KA
4- + 4- KA
+ 4- » HA
+ - KA KA
+ - KA HA
+ + 4- KA
+ 4- 4- XA
4 4- + SA
4- - HA KA
4- 4. + SA
+ - KA XA
* - HA HA
-* - HA RA
-+ 4- 4- .+
* ,+ - 4- ^HA
4- 4> " 4> H-
+ + .--+ KA
+' + * HA
mmi iugj**c prob
5
HA
*
KA
*
KA
KA
*
SA
KA
HA
HA
*;
t.:
^^^i
ably
6
KA
KA
HA
HA
KA
KA
HA
KA
KA
HA
KA
KA
HA
HA
HA
KA
KA
SA
KA
KA
HA
KA
4- .
SA
* - -
S*
4>
HA
4-
KA
^.^^
ooe
Av«rag*
Total local
Judgement TOJD TCSD !
Pos. Defl KD (ppb) (?pb) :
^ 3.6
/ KD
<* 39
f 45
/ 23
/ 6
/ 74
V* 43
/ 7
/ SD
/ 236
/ 126
/ 274
/ 50
/ ' 83
/ 79
/ KD
/ KD
«* KD
/ 56
/ 124
/ 82
/ 3270
/ 3984
/ 5.7
/ HD
/ 9.6
/ KD
-J HD
/ 2.2
/ HD
/ - 2230
f- -^ 83
^ 57
- ' J "16
' J '. 13
/
* 9.6
f 7.1
*f - 1610
^ 1732
dioxin- coacmtinaat or lotarferae*
KD
Defl.j
Defl.
Btfl.
KD
212
81
HD
87
3627
HD
HO
HD
Defl.
Dtfl.
XD
Defl.
MMB^BM
[«
33
-------
TABLE 14. VARIATION OF TCDD DETECTION. JUDGEMENT VERSUS EPA SAMPLE TYPE
Sample Type
Number Batch
Samples Analyzed
Number/(Percent) of Batch Samples
Versus Detection Judgement
Positive
Doubtful Nondetectable
2,4,5-Trichlorophenol
2,4,5-T
Butoxyethyl Ester of 2,4,5-T
Isooctyl Eater of 2,4,5-T
Butoxypropyl Ester °f Silvex
; Isooctyl Ester of Silvex
, Chlorinated Phenolic Antiseptic
Organophosphate
TOTALS
! Twenty different QG
Samples Assayed
16
10
3
7
22
_7
69
AUTHENTIC PESTICIDE BATCH SAMPLES
l/(33.3)
9/(90)
6/(86)
17(25)
6/(30)
l/(33.3)
I/(10)
3/UOO)
167.100^
W(33.3)
22/(100)
7/(100)
QUALITY CONTROLS
6/(30) : 8/ (40)
-------
TABLB 15. VARIATION Og TOTAL TCDD LEVEL VERSUS SAMPLE TYPE FOR POSITIVE SAMPLES
TCDD Range (ppb)
Sample Type
Number Batch Samples
Positive for TCDD
100- 500- 1000- 1500- 2000- 2500- 3000- 3500
10-30 31-99 499 999 1499 1999 2499 2999 3499 3999
Number Batch Samples/(Percent)
; 2,4,5-T
i : . :".'-; ;
Butoxyethyl
Butoxypropyl
Silvex
:-' : -,< -.
s . . '
'' «/.C^'«;i: '*';- i '
Ester of;
Ester of
Isooctyl Ester of
Silvex '
1
-i '
' !,".9
6
1
AUTHENTIC PESTICIDE BATCH SAMPLES
1/(18)
9/(100) ,
6/(100)
I/ (100)
QUALITY CONTROLS
Six different QC Samples
Positive for TCDD .
2/(32.8) 1/U6.8)
-------
TABLE 16.
LJl
0\
Statistical. Analysis
Total TCDD Concentration (ppb)
Sample
1-01
11
3Y
3X
4
14-01
3Z ..'
222841 .
222455
220777
226617
225511
229617
90
303
304
305
Run No, 1
236
80
ND
56
3270
5.7*
./.' .''ND
; 2.2a
3,6
39
23a
,74
""" 7a
85
, 16a
9.6
1610
Run No. 2(-D)
126
63
ND
124
3984
ND
ND
ND
ND
45
6a
43
ND
57a
18a
17.1
1732
Run No. 3(-T)
274
79
NA
82
NA
9.6a
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
Average
(ppb)
i 212
81
NA
87
3627
i
i
42
15
59
1
'. 71
' 17
1 8.4,
1671
Standard
Deviation
(ppb)
77
2.1
NA
34
504
4.2
12
22
20
1.4
1.8
86
Relative Standard
Deviation
(*)
36
. 2.6
NA
39
14
;
9.4
83
37
.
28
8.3
21
5.2
Qualitative criteria Judgement; TCDD detection doubtful.
-------
TABLE 17, COMPARISON OF REPLICATE RESULTS FOR AUTHENTIC PESTICIDE BATCH SAMPLES
1
Total TCDD Concentration (ppb)
Type
Sample
2.4,5-T
Chlorinated Phenolic
Antiseptic
Isooctyl Ester of
2,4,5-T
Butoxyethyl Ester
of 2,4.5-T
.
Butoxypropyl Eater
of Silvex
i .
Organophosphate
Formulations
Batch
Sample
137402
137403
137832-3
137832-4
137832-5
1419968
141998-1
148919-1
148919-2
14B919-5
148919-6
148919-7
148919-8
148919-9
138919-10
148920-2
148920-3
148920-4
148920-5
148920-7
148921-3
148921-4
Run No,
1
17
14
ND
ND
ND
15
28
23
25
22
28
33a
14
23
27
19
21
20a
24
22
ND
ND
Run No. Run No. Run No.
2(-D) 3(-T) 4C-Q)
18
ND
ND
ND
ND :
30 21 27
13a
19
21
23
22 21
19a
30
23 19
19
17 22
17
20a
17 23 I
21 ;
ND
14a
Statistical Analysis
Average
(ppb)
IB
23
21
21
23
23
24
26
22
22
23
19
19
20
21
22
Relative
Standard Standard
Deviation Deviation
(ppb) (%)
.0.71
7
11
2.8
2.8
0.71
3.8
9.9
11
2.3
5.7
' 2.5
2.8
_
3.8
0.71
4
30
52
14
13
3.1
15.8
38
51
10.3
25
13.2
15
18.1
3.3
Qualitative criteria judgementt TCDD detection doubtful. ,
A third sample of 148921-3 and 148921-4 was also run but not reported due to a 0 percent recovery.
-------
Appendix A. Data Sheets for Sample 148919-5-2
38
-------
' ANALYSIS OF TECHNICAL GRADE PESTICIDES FOR TCDD
Data Sheet
Sample No. lid919-5-3 Date Received e/a/7* Date of GC/MS Analysis 8/e/r&
Date and Type Of Extraction /ntonnnp 7//8/T& / Mex#A>& - A/a.0//-
Sample Weight /-Qos^j . Sample Volume £M£ Total Extract
GC Conditions : Column 3' OV-//;3'7o Injector Temperature
Carrier Gas and Flow Rote ..^f^ . A/
-------
ANALYSIS OF_TECHN1CAL GRADE PESTICIDES FOR
/*&3/&-3'&-a TCDD
Results
RECOVERY
37C1-TCOD Std. Conc.(pg//il) * ₯80 % Recovery (R)
Added to Sampling) * #8 ( I ) R _ _H_X j 0 0 -
Recovered (ng):
i/s xJg.»
QUANT1RCAT10N
35C1-TCDD Std. Conc.(pg//tl) -
520 522
Amount in Extract (ng) « i/s x ^2 ^-3 /^^ (IEL)
Amount in Sample (ng)=m/R 22- g ^^7 (IZD
Sample Conc^ppb)= IZ/g Sample
Avg.Conc.(ppb) =
LIMIT OF DETECTION
N/A A
L.D.,2.5(S/N)
CONFIRMATION ANALYSIS (Fragmentation Pattern Recorded)
N/A _ - A
Result Positive
Result Negative
-------
.Appendix B. Master Sample Status Form
41
-------
Master Sample Status For 328-884-11
Sample
Recovery ,%
Screening
.
Repeat
'
Average
TCDD
Level
(ppb)
Full Scale
Configurations
Needed
'
Done
Calculated
LD
(ppt)
.
Sample
Completed
page
-------
Appendix C. Cumulative Tabulated Analytical Data Form
. .43
-------
CUMULATIVE
TABULATED ANALYTICAL DATA FOR 328-884-11
Sample
Recovery, %
Screening
Repeat
Average
TCDD
Level
Ippb)
.
Confirmation
Calculated
LD
(ppb)
i
Sample
Completed
GSRI
EPA
Code: NA-notapplicable ND-not detectable
v* -sample completed
-task performed
Page.
-------
Appendix D. GC/MS Performance Ion Current Cnromato grams
(Observation #21)
OBSERVATION £21 DATA
Ul eo-injeeted m/e/Paak area
/
TCDD 37d TCDD 320 322 328
2+2
1 + .1...
2 "₯ 2 ~" "
1*1
Corr (r)
Slope (s)
Intercept (i)
s/500 or 480
' 1257
' 583
1203""
576
0.9999
680.5
-101
1.36
1560
735
"' ~" 1612 ~
722
0.9990
857.5
-129
1.71
2198
1073
~ ~~ 2317" ""
1176
0.9952
1133
-8.5
2.36
Avg. 320/322 - 0.795
45
-------
** SELECTED ION CHROMATOGRAM ##
HUL TCDD HATIUE CL35 +CL37 STDS CHECK
31JULY78, OV11, 1 OF 4
FRN 31079
1ST SC/PG- 1
X- 1 00 Y- 1.00
337.:
19.3t=£=
A- 8198.
A- 1560.
1357.
-LA
-------
** SELECTED ION CHFOriATOGRAH ** ! FRN 31080
1UL hATIUE TCDD CL35 + 1UL CL37 STDS CHECK 1ST SC/PG- 1
31JULV78, OU11. 3 OF 4 X- 1.00 V- 1.00
-------
** SELECTED ION CHRGMATOGRAH **
TCDD NATIUE CL35 +CL37 STDS CHECK
3iJULV?S. OVil, 3 OF 4
FRN 31081
1ST SC/PG- 1
X- 1.00 Y- 1.00
Co
337.3
321. G
19.:
3317.
161B
1E63.
fi_
-------
** SELECTED IOH CHROMATOGRAM **
lUL TCDD NATIC'E CL3S +CL37 STDS CHECK
31JULV78, OU11, 4 OF 4
37
31.
is.:
FRN 81083
1ST SC/PQ- 1
X- 1.00 V- 1.00
4 fl
-------
^.Appendix E. Ion Currrent Chromatograms for
TCTD Detected la a Pesticide Sample
(ZPA Sample if4)
QUALITATIVE CZTTZSIA
123
1 + + -i-
4
: H-
5
+
6
-
JITDGEMEHT: TCDD detected in Che sample
because ail siz criteria wars positive.
50
-------
FAN ft1604
1ST 0C/PQ- I
x- i.e» v
a+
1BAU078, KMRO
TCDD Retention Time
-------
'01
** SELECTED ION
-------
Appendix F. Mass Spectrum for TCDD Detected -
in a Pesticide Sample
(SEA Sample #143919-5)
HOTS: Existence of m/e lines at 320, 322, 324 and 326
37
' indicative of line at m/e 328 due to Cl TCDD.
53
-------
Ut '
DORK AREA SPECTRUM FRfl 21193 . PAGE 1
LARGST 4- 327.9,100.0 321.8. 43.0 319.9, 34.3 393.
LAST 4' 333.9, 31.0 325.8, 16.0 337.9.100.0 328.
-43 + 53 -58
I O0
80
60
40
30
0
100
80
60
40
30
0.
V - 1.00
0, 39.3
9, 15.1
30 40 60 80 100 130 140 160
,
ll li
, , .., i i ,,.,. Jin , 1 1
18ft 300 320 340 360 380 30
,
0 3f?0>
-------
01
u<
UORK AREA SPECTRUM FRII 31193
LARGST 4« 337.9,100.0 321.8. 43.8
LAST 4- 333.9, 31.0 32S.8. 16 0
-4S + 53 -68
PAGE 3 V - 1.00
319.9, 34.3 B93.0, B9.8
337.9.100.0 338.9. 15.1
1001
80]
i
60
40
80
0
100]
80]
60]
40]
80]
0.
ll
MIII iiM)innimiiu-iimmniimiii iimmi niiiun iiiiitiii-iniiim imiirn iiiiiiiiriiiiiMiriiniiririininii
340 360 380 400 420 440 460
-im|
480
500.
-5SQL
IIIIJIHI
nti]
-------
Appendix G. Mass Abundance Tabulation for TCHD
Detected In a Pesticide Sample
(EPA Sample #90)
HOTZ: Mass abundances at m/e 320, 322, 324 and 326
indicative of TCED.
56
-------
Ln
FRN 31643 .SPECTRUM 47
RET. TIME
4.7
> PAUSE
MASS
347
243
249
350
251
asi
352 '
853
354
255
ass ;
257
859
360
B61
£62
863
B64
864
864
865
866
867
868
869
370
ABUND
IS.
11.
19.
11.
' ,.. . 15.
15.
8.
45.
ea.
: 40,
15.
.;":.,. 59.
473 .
130.
41,
21.
28.
8.
7.
7.
. ' i 9. i
8.
19.
10.
15.
7.
MASS
371
273
273
374
375
376
377
878
279
880 '
381
383
383
384
385
287
888
389
890
391
393
393
394
895
396
ABUND
8..
11.
14.
135.
81.
33.
11.
8,
8.
6.
40.
13.
IS.
6.
.: 13.
13.
26.
11.
86.
15.
11.
31.
8.
7.
4.
MASS
397
£97
398
299
300
300
301
302
303
304
305
306
307
307
30*
309
310
311
313
313
314
315
316
317
318
ABUND
MASS ABUND
4.
3.
4.
7.
3.
3.
5.
5.
7*
5.
4.
8.
11.
10.
.9.
13.
5.
7.
4.
9.
6.
45.
19.
53.
18.
319
319
320
321
323
324
335 ,
326
338
329
330
331
332
333
334
334
336
336
337
337
338
339
340
341
343
7.
7.
32.
10.
89.
14.
9.
7.
38.
10.
5.
15.
11.
9.
5.
4.
4.
4*
6.
6.
6.
4.
a.
7.
4.
-------
Appendix H. Ion Current Chromatograms for
Nondetectable TCDD in a Pesticide Sample
(EPA Samples #137834-3 and. 3TD)
QUALITATIVE CRITERIA
Sample 1 2
137834-3 +
.. 3CT ». - ..--.- + - -
3
HA
- HA
4
HA
HA
5
HA
HA
6
HA
HA
JUDGEMENT: TCDD not detected- in the sample because criteria #2
was negative due to absence of both m/e 320 and 322 peaks.-
58
-------
** SELECTED ION CHROflATOGPfiM **
2'800UL PEST EXTR TODD 137334-3 + OULS STANDARD
05JUH7S. OU-li. 210(1 >240f6>, 1 OF A
FRN
1ST SC/PG-
X- 1.00 V- I.00
18783,
Ul
V0
119,
-------
o\
o
** SELECTED ION CHPOMnTOGRAfl *# FRN
2'2C»OUL TCDD PEST EXTR 137834-3 4- SUL CL35 + 1UL CL37 1ST 5C/PG- 1
05JUM7S, OU 11. 210(1 )S40(6>. 2 OF 4 X" 1.00 Y- l.OO
337
321.
119
38707.
A- 35323.
19558.
-------
cr>
t* SELECTED IWI CHRCtlHtOORAH ** FKN
EXT 3v-D-a + 3 BUL $OLV r«> STDS aoffPT7» ICT «C/PQ
206EPT78 OV1I BlOit»26E«lO X- t.OO V-
1 OF 4
187,
331,
19'
IE
Cl TCDD
Extraneous peaks overlap with TCDD;
therefore, not possible to determine
native TCOD.
A*4353
A=6626
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** SELECTED I*N CHPOJKVTOGRArt *» FRN 81354
PEST EXT 3V-D-B + BUL CL37 tl UL CL3S TCDD *eSEt»T»SCxFO 1
ooit 3ioa>E6a*io x- i.eo v
8 OF 4
TCDD Retention Time
ft- 60371.
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Appendix I. Ion Current Chronatograins for Questionable
TCDD Detection in a Pesticide Sample
; (ZPA Sample #90D)
QUALITATIVE CRITERIA
! 1 ' 2
.4- +
3 ' 4 5
+ NA
6
NA
JUDGEMENT: TCDD detection questionable
because criteria Z?5 was negative due to
m/e-ratio out-of-specification.
63
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i Ch
** SELECTED I OH
B^SeOUL, PEST EXTR »d-D-8 * 6UL3 STD
01SP78, HP6886. QUll. 810U >2o
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.; I
o\ .
Ol
** SELECTED ION CHROMATOGRAM **
a/aoouL PEST EXTR SO-D-S + IUL etas + aut CL37
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