United States
Environmental Protection
Agency
Environmental Monitoring and
Support Laboratory
Cincinnati OH 45268
Research and Development
EPA/600/S4-85/029 May 1985
Project Summary
Development of Methods for
Pesticides in Wastewaters
J. S. Warner, T. M. Engel, and P. J. Mondron
The project covered by the full report
involved the development of monitoring
procedures for the analysis of 33 pesti-
cides in relevant industrial waste waters.
The procedures were modeled after
existing 304(h) procedures, specific yet
as simplified as possible. Common
techniques such as packed column gas
chromatography (GC) and high perform-
ance liquid chromatography (HPLC)
with specific detection systems were
used in conjunction with standard
work-up procedures based on separa-
tory funnel-methylene chloride extrac-
tion. Kuderna-Danish (K-D) concentra-
tion, and silica gel or Florisil cleanup
whenever possible. Method detection
limit (MDL) and the applicable analytical
range were determined from spiked
reagent water samples. Precision and
accuracy of the methods were deter-
mined in relevant industrial wastewaters
where possible.
This Project Summary was developed
by EPA's Environmental Monitoring
and Support Laboratory, Cincinnati,
OH, to announce key findings of the
research project that is fully docu-
mented in a separate report of the same
title (see Project Report ordering infor-
mation at back).
Introduction
The methods described in the full report
were developed to meet USEPA's respon-
sibility to promulgate multi-residue test
procedures for the analysis of pesticides
in industrial wastewaters. This protocol
was modeled after the form and philos-
ophy of existing and proposed 304(h)
methods. Common techniques and equip-
ment were employed and although speci-
ficity was required, the procedures were
kept as simple as possible with a minimum
number of steps. The preferred determin-
ative step was gas chromatography; high
pressure liquid chromatography was the
second choice.
The compounds investigated were cat-
egorized according to:
• Similarities in structure
• Similarities in determinative step
• Co-occurrence in relevant industrial
wastewater
The protocol had the following goals:
• 1 f/g/L gas chromatographic detection
limit or 10#g/L HPLC detection limit,
and
• 85 percent recovery of analytes from
reagent water.
Chromatography
The first step in development of each
test procedure was the experimental
determination of a detection system that
met the sensitivity requirement for each
one of the compounds to be analyzed in a
given group. Chromatographic columns
and operating parameters were selected
that would separate the compounds
expected to co-occur in a given industrial
waste sample. The separation had to be
sufficient for quantitation while keeping
the analysis time under 1 hr. In some
cases, more than one system was re-
quired for a given wastewater. This
occurred when a single industrial facility
manufactured either structurally very
similar or very dissimilar compounds.
Problems requiring the development of a
second chromatographic system were
coelution, retention times of more than 1
hr, and inadequate sensitivity for one or
more compounds produced at a single
site.
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Model Verification or
Modification
After chromatographic conditions were
selected, the applicability of the model
procedure suggested by existing 304(h)
methods was tested for each compound.
The model utilized 1 -liter sample volumes
extracted with three aliquots of methylene
chloride followed by K-D concentration
and standard column cleanup techniques,
if required.
Extraction and Concentration
The percent recovery of each compound
from deionized water at neutral pH was
determined, and adjustments in the pH of
the water or the volume of the extraction
solvent were used to maximize the per-
cent recovery value of this step.
Cleanup
The appropriate solvent fortified with
each compound was applied to a standard
adsorbent column and the column eluted
with various solvent mixtures. The elution
pattern and percent recovery value for
this procedural step were determined for
each compound. Variations from the
cleanup step of the model procedure
included changes in the composition of
elution mixture, water deactivation of the
sorbent, and use of various sorbents.
Often, further method modifications were
required to separate the compounds of
interest from interferences found in the
relevant wastewater. The systems devel-
oped and described in each report must
be considered only as a starting point in
effecting an adequate cleanup.
Application to Relevant
Industrial Wastewater
After extraction and cleanup proce-
dures were developed using fortified
reagent water, industrial wastewaters,
which reflected the matrices in which the
studied compounds were likely to be
present, were analyzed. Further modifi-
cations were made as necessary. Selected
compound groups were then fortified at
detectable levels into relevant industrial
wastewater samples and percent recovery
values from this matrix were determined.
Precision and accuracy were deter-
mined from the analysis of seven repli-
cates of fortified relevant industrial
wastewaters. Method detection limits
were determined in fortified reagent
water samples. Analytical curves were
generated to determine and demonstrate
the linearity of recovery of the compounds
from reagent water.
Table 1 lists the methods developed
and the compounds within each method.
The compounds within a method desig-
nation are grouped according to similar
analytical characteristics.
Table 1. Pesticides Grouped by EPA
Method Numbers
604.1 Phenolic Pesticides
Dichlorophen
Hexachlorophen
616 Neutral C. H & O Pesticides
Cycloprate
Kinoprene
Methoprene
Resmethrin
618 Volatile Pesticides
Chloropicrin
Ethylene dibromide
620 Diphenylamine
622.1 Thiophosphates
Dichlorofenthion
Famphur
Fenitrothion
Fonofos
Phosmet
Thionazin
633.1 Neutral Nitrogen Containing Pesticides
Fenarimol
MGK - 264
MGK - 326
Pronamide
634 Thiocarbamate Pesticides
Butylate
Cycloate
EPTC
Moilnate
Pebulate
Vernolate
Giv Card
635 Rotenone
636 Bensulide
637 MBTS and TCMTB
638 Oryzalin
639 Bendiocarb
640 Mercaptobenzothiazole
Table 2 lists some chromatographic
systems and parameters utilized in the
methods. Stability studies (holding times,
etc.) were not specified in this contract.
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Table 2. Chromatographic Systems and Parameters Developed for Methods
MDL Chromatographic
Method Compound (ug/L) Detector Column
604.1
616
618
620
622.1
633.1
634
635
636
637
638
639
64O
Dichlorophen
Hexachlorophen
Cyc/oprate
Kinoprene
Methoprene
Resmethrin
Chloropicrin
Ethylene Dibromide
Diphenylamine
Aspon
Dichlorofenthion
Famphur
Fenitrothion
Fonofos
Phosmet
Thionazin
Fenarimol
MGK-264
MGK-326
Pronamide
Butylate
Cycloate
EPTC
Molinate
Pebulate
Vernolate
Rotenone
Bensulide
MBTS
TCMTB
Oryzalin
Bendiocarb
Mercaptobenzothiazole
1.0
1.2
21
18
22
36
0.8
0.2
1.6
0.6
0.7
19
2
0.7
1
1
4
2
6
4
0.6
1.6
0.9
0.6
0.8
1.1
1.6
1.6
0.5
1.0
0.5
1.8
1.7
UV/254
FID
FID
FID
FID
ECD
ECD
NPD
NPD
NPD
NPD
NPD
NPD
NPD
NPD
NPD
NPD
NPD
NPD
NPD
NPD
NPD
NPD
NPD
NPD
UV/254
UV/270
UV/254
UV/254
UV/254
UV/254
5 u. Spherisorb-ODS,
250 x 4.5 mm
3%SP-2250on100/120
mesh Supelcoport
1%SP-1 000 on Carbopak B
f 60/80 mesh), 1.8 m x 2 mm
ID, glass column
3% SP-2250 on Supelcoport
1 100/120 mesh). ISOcmx
2 mm ID, glass column
3% SP-2250 on Supelcoport
(100/200 mesh) 1.8 mx 2 mm
ID, glass column
1 3% SP-2250 on Supelcoport
(100/200 mesh) 1.8 mx
2 mm ID, glass column
3% SP-2100 on Supelcoport
(100/1 20 mesh) 1.8 mx
2 mm ID, glass column
5uZorbax-CN
250x4.6 mm
Su, Spherisorb-ODS,
250 x4.6 mm
Su, Dupont Zorbax-CN.
250 x4.6 mm
Su, Spherisorb-ODS,
250 x 4.6 mm
5fj. Spherisorb-ODS,
250 x 4.6 mm
Su, Spherisorb-ODS,
250 x 4.6 mm
Operating
Parameters
65/35 acetonitrile/water .05%
HsPO4 at 1 mL/min
Programmed from 180° to
240° at 8°/min,
Injector - 280
Det. = 300°
1 35° Isothermal
Column temp, held at 80° for
4 min programmed to 300°C at
8°/min and held at 300° for
4 min
Column temp, held at 80°C for
4 min and programmed to
300°C at 8° /min and
held at 300° for 4 min
Column temp, at 80° for 4 min
and programmed to 300° at
8° /min and held for 4 min
Column temp, at 80°C for 4 min
programmed to 300° at
8° /min and held for 4 min
30/70 methylene
chloride/ hexane, at 1 mL/min
55/45 acetonitrile/water.
1 mL/min
15/85 methylene
chloride/ hexane, at 1 mL/min
40/60 acetonitrile/water
at 1 mL/min
40/60 acetonitrile/water
at 1 mL/min
50/50 acetonitrile/water
at 1 mL/min
Retention
Time (min.)
4.2
9.7
3.6
4.4
5.5
8.4
5.6
9.9
18.1
22.6
21.4
28.1
23. 1
20.5
30.0
18.3
30.6
23.0
21.9
19.9
18.2
24.2
17.9
23.8
20.2
19.6
8.6
14.1
6.6
9.3
6.2
9.3
8.4
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J. S. Warner, T. M. Engel, and P. J. Mondron are with Battelle Columbus
Laboratories, Columbus, OH 43201; the EPA author Thomas A. Pressley (also
the EPA Project Officer, see below) is with the Environmental Monitoring and
Support Laboratory, Cincinnati, OH 45268.
This Project Summary covers 13 separate reports, entitled:
"Determination of Thiophosphates in Industrial and Municipal Wastewaters—
Aspon, Dichlorofenthion, Famphur, Fenitrothion, Fonofos, Phosmet, and
Thionazin," (Order No. PB 85-189 017/AS: Cost: $10.00)
"Determination of Thiocarbamates in Industrial and Municipal Wastewater—
Butylate, Cycloate. EPTC, Molinate, Pebulate, and Vernotate,"(Order No. PB
85-189 009/AS; Cost: $10.00)
"Determination of Neutral Nitrogen-Containing Pesticides in Industrial and
Municipal Wastewaters—Fenarimol, MGK 264, MGK 326, and Pronamide,"
(Order No. PB 85-189 157/AS; Cost: $10.00)
"C, H, and O Compounds—Cycloprate, Kinoprene. Methoprene, and Res-
methrin."(Order No. PB 85-188 951/AS; Cost: $10.00)
"Determination of Volatile Pesticides in Industrial and Municipal Wastewaters,"
(Order No. PB 85-189 918/AS; Cost: $8.50)
"Determination of Bendiocarb in Industrial and Municipal Wastewaters,"
(Order No. PB 85-188 159/AS; Cost: $8.50)
"Determination ofBensulide in Industrial and Municipal Wastewaters, "(Order
No. PB 85-188 9777AS; Cost: $8.50)
"Determination ofRotenone in Industrial and Municipal Wastewaters." (Order
No. PB 85-188 993/AS; Cost: $8.50)
"Determination of Oryzalin in Industrial and Municipal Wastewaters," (Order
No. PB 85-188 985/AS; Cost: $8.50)
"Determination of Diphenylamine in Industrial and Muncipal Wastewaters."
(Order No. PB 85-190 874/AS; Cost: $8.50)
"Determination of Mercaptobenzothiazole in Industrial and Municipal Waste-
waters." (Order No. PB 85-191 120/AS; Cost: $8.50)
"Determination of Hexachlorophene and Dichlorophen in Industrial and
Municipal Wastewaters."(Order No. PB 85-188 209/AS; Cost: $8.50)
"Determination of MBTS and TCMTB in Industrial and Municipal Waste-
waters," (Order No. PB 85-189 025/AS; Cost: $8.50)
The above reports will be available only from: (cost subject to change)
National Technical Information Service
5285 Port Royal Road
Springfield. VA22161
Telephone: 703-487-4650
The EPA Project Officer can be contacted at:
Environmental Monitoring and Support Laboratory
U.S. Environmental Protection Agency
Cincinnati, OH 45268
U.S GOVERNMENT PRINTING OFFICE: 1985-559-016/27
United States
Environmental Protection
Agency
Center for Environmental Research
Information
Cincinnati OH 45268
Official Business
Penalty for Private Use $300
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