United States
Environmental Protection
Agency
Environmental Monitoring
Systems Laboratory
Las Vegas NV 89193-3478
Research and Development
EPA/600/S4-89/007 Sept. 1989
&EPA Project Summary
Biological Monitoring
Systems for Hazardous Waste
Sites (Production and
Analysis of Analytical
Reference Materials)
V. R. Bohman, C. R. Blincoe, G. C. Miller, R. L. Scholl, W. W. Sutton, and
L. R. Williams
Portions of EPA programs in
pesticides, toxics, and hazardous
waste have a need for various types
of analytical reference material. The
current project has emphasized the
collection and analysis of urine, fat,
and blood from cattle exposed to
selected toxicants for ultimate use as
reference samples. The project also
addressed the practicality of using
certain metabolites produced by the
cattle to indicate previous exposure
to chlorinated hydrocarbons and the
likelihood that these metabolites
could serve as conditions of expo-
sure even if an animal had ingested
small amounts of many other chem-
icals such as might be present in the
vicinity of an uncontrolled hazardous
waste site.
The reference samples of urine, fat,
and other tissues can, with verified
compound concentrations, ultimately
be used as qualifying samples when
selecting an analytical laboratou
from among several candidates jrj
when selecting the best technique'to
use for a particular analysis. How-
ever, the reference materials may be
of greatest benefit when used by
laboratories to determine analytical
accuracy for samples of human
urine, blood, etc. This is because the
standards, like the unknown samples,
will contain pollutant compounds and
associated metabolites (all In vivo
incorporated) in an appropriate te»o-
logical matrix. <
Dairy animals were used during
this study primarily because substan-
tial quantities of tissues and body
fluids could be collected from the
previously dosed cattle. WhMe none
of (he individual study phases
included a thorough evaluation of
doslnq compound retention, excre-
tion, and/or metabolic degredation,
each animal was given daily oral
doses of selected toxicants so that
the urine, blood, etc., would contain
detectable concentrations of the
dosing compound(s) and some of the
associated major metabolites. Aliq-
uots of the respective large-volume
samples were chemically analyzed
and, based on the initial analytical
results, some of the samples were
then selected, as potential reference
materials. Several confirming anal-
yses (not part of this specific project)
wilj, ultimately be conducted before
any sample Is actually distributed as
an analytical reference or quality
assurance material.
The project report contains several
data summaries organized by individ-
ual animal and by individual dosing
compound. Analytical and sample
preparation steps are discussed and
the analytical detection limits are in-
cluded for each dosing compound
and for each sample type. Some of
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the samples provided by this study
have already been used in an on-
going EMSL-LV program for analytical
method optimization.
This Project Summary was devel-
oped by ERA'S Environmental Monitor-
ing Systems Laboratory, Las Vegas,
NV, to announce key findings of the
research project that is fully docu-
mented in a separate report of the
same title (see Project Report order-
ing information at back).
Introduction
Assessing current and potential prob-
lems at uncontrolled hazardous waste
sites has been very difficult due to
several factors including (1) the complex
chemical mixtures found at most sites
and (2) the fact that toxicity and
environmental transport data are limited
for many of the compounds present at
the site, especially for those that are by-
products of organic synthesis (rather than
commercial chemicals). Similarly, when
samples of urine, blood, milk, etc., are
collected and analyzed in an attempt to
assess exposure and uptake, it is very
difficult to determine the accuracy of
these analyses due to a lack of adequate
reference materials. Under conditions of
mammalian exposure, many of the
pollutants would be partially metabolized
and several conjugation or degradation
products would be present in a collected
sample. The currently available standards
are usually prepared by adding the
pollutant compound(s) to an uncon-
taminated sample of biological tissues or
fluids. While this approach provides a
reference compound in the correct
sample matrix, it does not include the
array of associated compound degrada-
tion products, many of which can
interfere with an accurate analytical
determination and some of which may
have high toxicological significance. Por-
tions of EPA programs in pesticides,
toxics, and hazardous waste have a
defined need for analytical reference and
quality assurance materials.
This study was conducted to determine
(confirm) the retention and excretion
pattern of selected hazardous waste site
chemicals and to provide analytical
reference and quality assurance samples
that contain both a representative con-
centration of the pollutant compound(s)
and representative concentrations of the
associated metabolic degradation prod-
ucts. Cattle were used as experimental
animals throughout the study (1) because
of their potential selection as a biological
monitoring species for chemical pollu-
tants in agricultural areas, (2) because
they provide major food products for
human consumption (i.e., milk and beef)
and, perhaps most importantly, (3)
because substantial quantities of mam-
malian tissue and body fluid could be
collected and prepared for subsequent
use as reference materials. In each phase
of the project, cattle were given daily oral
doses of compounds that are of regu-
latory interest to one or more of the EPA
program offices. Samples of urine, blood,
milk, feces, and sacrifice tissues were
subsequently collected and portions of
the respective samples were analyzed
using computer assisted gas chromatog-
raphy/ mass spectrometry. The selection
of chlorinated hydrocarbons, emphasized
in phases I and II, was based on results
from a 1980 Love Canal (Niagara Falls,
New York) monitoring program and on a
preliminary survey of toxic chemicals
frequently present in hazardous wastes.
During the first part of this project,
some brief method evaluation tests were
conducted to select the specific analytical
and sample preparation procedures that
would be used during subsequent study
phases. Another objective of the initial
effort concerned the practicality of using
certain metabolites to indicate previous
exposure to chlorinated hydrocarbons
and whether these metabolites could be
used if an animal had ingested not only
chlorinated hydrocarbons but had also
ingested small amounts of many other
waste site chemicals. Indicator com-
pounds would include those compounds
and metabolites that, when detected in
urine, blood, etc., suggest previous expo-
sure to chlorinated hydrocarbons.
The second part of the overall project
(and the phase for which there is a
continuing EPA program office interest)
emphasized the collection and analysis of
blood, fat, and urine samples for ultimate
use as analytical reference materials.
Substantial quantities of body fluid were
easily obtained from the dairy animals.
Each animal was given daily oral doses
of selected pollutant chemicals so that
the tissues and fluids, would contain
detectable concentrations of the dosing
compound(s) or associated major metab-
ttes. Aliquots of the respective large
nume samples were chemically ana-
lyzed and, based on these initial ana-
lytical results, some of the samples were
then selected as potential reference
samples. Confirming chemical analyses
will ultimately be conducted before final
sample selections are made and before
any sample aliquots are distributed as
reference or quality assurance materials.
The reference samples, with ver
compound concentrations, can be
as performance evaluation samples v
selecting an analytical laboratory
among several candidates or w
selecting the best technique to use I
particular analysis. However, the r<
ence materials may perhaps b<
greatest benefit when used by lab
tories to determine analytical accui
for samples of human urine, blood,
As mentioned previously, this is bec<
the standards, like the unknown samr.
will contain pollutant compounds
associated metabolites (all in vivo in
porated) in an appropriate biolog
matrix.
Sample Collection
Dosing groups for the four indivic
study phases are shown in Tables 1, 2
and 4, respectively. No dosing adji
ments were made for individual variatii
in animal weight. However, individ
animal weights were taken before dos
began and at time of sacrifice. All anim
received a commercial feed preparat
and had continuous access to water fr
individual automatic watering units.
In phase I, dairy cows were on
dosed with a mixture of chlorinal
hydrocarbons (2,4-dichlorophenol, 1
dichlorobenzene, lindane, 1,2,3,4-tet
chlorobenzene, and pentachlorophem
Selected tissues (e.g., liver, kidnc
muscle, fat, and blood), milk, urine, a
feces were analyzed for both the pare
dosing compounds and for some of t
major metabolic conjugation/ degradati
products. During phase II, eight anim;
received various combinations of tl
phase I chlorinated hydrocarbons and
liquid dose that had originally be<
collected from an actual hazardous was
site. While the waste site liquid was n
toxic at the amounts administered, it d
contain a broad array of organic cor
pounds. Large volume samples of urii
were retained for possible use i
analytical references and, as in phase
all collected sample types were analyze
for the chlorinated hydrocarbon dosin
compounds and for some of the maj<
metabolic degradation products.
Phase III and phase IV were conducts
strictly to provide samples for ultima!
use as analytical reference and qualil
assurance materials and, in some ir
stances, to provide sample material fc
use in a continuing EMSL-LV program (
analytical method optimization. The divei
sified group of phase III and phase I'
dosing chemicals included a carbamalj
insecticide, some polycyclic aromati*
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Table 1. Chlorinated Hydrocarbon Doses Administered to Phase I Dairy Cattle
Dose Per Cow
Dosing Groups
Daily dose per
animal
Total dose for
28 days
mg/kg/day
(approximate)
Two Animals (Group I)
lindane
Two Animals (Group II)
lindane
1,2 -dichlorobenzene
2,4-dichlorophenol
1,2,3,4-tetrachlorobenzene
Two Animals (Group III)
lindane
1,2-dichlorobemene
2,4-dichlorophenol
1,2,3,4-tetrachlorobenzene
pentachlorophenol
4.0 grams 112 grams
2.0 grams
20.0 grams
20.0 grams
2.0 grams
1.0 gram
20.0 grams
20.0 grams
2.0 grams
1.0 gram
56 grams
560 grams
560 grams
56 grams
28 grams
560 grams
560 grams
56 grams
28 grams
6 mglkg
3 mglkg
29 mg/kg
29 mg/kg
2 mglkg
2 mg/kg
29 mg/kg
29 mg/kg
3 mg/kg
2 mg/kg
Note: Samples from this phase have already been used as part of the EMSL-LV
program for analytical method optimization (Marsden, PJ., EN. Amick, F.L
Shore, L.R. Williams, V.R. Bohman. and C.R. Blincoe. 1986. Characterization of
Bovine Urine and Adipose Interlaboratory Performance Evaluation Samples
Containing Biologically Incorporated Chlorophenols. Journal of Agriculture and
Food Chemistry. 34:795-800)
hydrocarbons, an ether, a phthalate, an
azobenzene dye, and an amme. Large
volume samples were again collected
from the animals, but at less frequent
intervals than in phase I and II. In
addition, less attention was given to the
identification of specific metabolic prod-
ucts that resulted from the respective
dosing compounds and, since the
materials of primary interest were urine,
blood and carcass fat, fewer types of
samples were collected.
Throughout the project, samples were
placed in Teflon containers and stored at
-23°C. Blood samples were collected by
jugular venipuncture using a 14 gauge
syringe needle which allowed the blood
to drain directly into the Teflon sample
container. Portable machine milkers were
routinely used during those times when
the milk collection was not retained for
chemical analysis. When milk samples
were collected for analysis, the animals
were milked by hand so that the milkwent
directly into the sample collection
container. Urine samples were typically
taken before dosing began and for the
first week of dosing. In some instances,
the cows were cathetenzed within
dwelling, inflatable urinary catheters and
the urine would drain through poly-
ethylene tubing into Teflon containers
located at the rear of each stall. When
tissue samples were collected, the
animals were sacrificed two days after
the last dose.
Sample Analysis
A continuing, though secondary, objec-
tive of this project was to improve the
applicability of currently available proce-
dures for the more difficult sample types,
i.e., those with a significant lipid content
such as milk, milk fat, carcass fat, and
liver. Some of the extraction steps were
modified primarily during the second
phase of the study, and additional proce-
dural revisions were made when ana-
lyzing for the more diversified group of
chemicals present in the phase III and
phase IV samples.
The amount of sample, or starting
material, used during an analysis was
typically 200 grams for urine and milk:
50 grams each for liver, muscle, kidney,
and blood: 10 grams for fat: and, 100
grams for feces and animal feed. The
sample analysis plan typically included
homogenization of the sample material,
enzymatic hydrolysis, solvent extraction,
cleanup and concentration of solvent
extracts, extract derivatization, and instru-
ment analysis. Samples of animal tissue,
feed, and feces were initially hortSo-
genized using a tissue grinder. With the
exception of carcass fat and animal feed
samples, the sample material was fre-
quently treated with a commercial
enzyme preparation to hydrolyze glucur-
onide and sulfate conjugation products.
The hydrolyzed samples were then ex-
tracted with organic solvents at high and
low pH to partition the base- neutral and
acidic compounds. Extracts were then
concentrated using either a Buchi
rotoevaporator or a Kuderna-Danish
apparatus. The acid fraction was then
derivatized to improve the stability of free
hydroxy compounds (i.e., compounds
resulting from the derivatization step).
The sample extracts were then analyzed
using computer assisted gas chroma-
tography/mass spectrometry. The
approximate detection limits that were
achieved during the study are shown in
Table 5 and a precision estimate, based
on the analysis of 10 aliquots of a single
urine collection, is shown in Table 6.
The specific analytical and sample
preparation steps that were used during
each phase of the study are included in
the project report. Dosing compounds
and a few of the metabolite compounds
(those that were commercially available)
were added to some of the samples in
order to allow for a subsequent estimate
of compound recovery. The resulting
compound recovery data are also pro-
vided in the project report.
Confirming Analysis
This study has provided many large
volume samples and aliquots oft these
samples have received an initial analysis
Consequently, some of the sampte*\ have
been tentatively selected as potential
reference material and the remaining
samples, collected as part of this overall
effort, have been discarded. Carcass fat,
blood, and urine represent the pre-
dominant number of sample types
retained.
The next step will be to assess the sta-
bility of compound concentrations under
typical storage conditions. This is ob-
viously a critical area and analyses will be
conducted at various intervals of time
and, perhaps, under different storage
temperatures. Another series of analyses
is planned to assess the variability in
compound concentration between ali-
quots of the same sample. The samples
would not necessarily serve as a
reference material for each in vivo incor-
porated pollutant compound and/or as-
sociated metabolite that might be present
m a given sample, but rather would be
used for one or more of the compounds
where extensive confirmation analyses
have been conducted. These confirming
analyses are frequently conducted by
more than one analytical technique.
Conclusions
None of the study phases was con-
ducted as typical metabolism experi-
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Table 2. Phase II Cattle that Received Doses of an Actual Hazardous Waste Site
Liquid as well as the Previously Administered Chlorinated Hydrocarbons
Liquid Material from Hazardous Waste Site"
Number of Chlorinated
Animals Hydrocarbon Dose" Dose/Animal/Day Total DoselAnimal
2
2
2
2
2
tratment A 80 ml
treatment A 160 ml
1/2 tratment A 160ml
80 ml
1120ml~"
2240 mr~
2240 ml"
2240 ml~"
"Chlorinated hydrocarbon dose (treatment A) composed of lindane - 1 gram/
animal/day; i,2-dichlorobenzene - 20 grams/animal/day; 2,4-dichlorophenol - 20
grams/animal/day; 1,2,3,4-tetrachlorobenzene - 2 grams/animalldays and pentachlo-
rophenol - 1 gram/ animal/day.
"Waste site material contained many semivolatile organics at mgli to figli
concentrations, i.e., benzaldehyde; benzoic acid; pentachlorophenol; 9,10-
anthracenedione; flouranthene; pyrene; p-phenylcarbamlic acid: 1 -methoxy-1 -
methyl9thO>v-2-propanol; 4-methoxy 2,2,6-trimethyl cyclohexanone; 4-hydroxy-3-
methoxy-benitldehyde; phenanthrene; anthracene; carbazole; biphenylene; 4-
methylphonol; 2-butoxyethanol; flourene; 1,2-benzenedicarboxylic acid, 1-
ethoxytwt*n»; trans-2-ohlorocyclohexanol; phenoxyacetic acid: 4-hydroxybenzene
ac«*c acid; mtthylcycloheptane; 5-methyl-l,2-haxadiene; 3-ethyl-i,4-hexadiene:
pftarMntfiridtorm: phenol: 2,3,4,6-tetrachlorophenol; 2-methylanthracene; dibenzofuran;
and M-acHytfwnzamide. Numerous inorganics (mg/1 concentrations) were also
present i.9, aluminum • 420; cadmium • 14; calcium - 200; chromium - 440; cobalt -
t30f copper - 710; iron - 470; lead - 3; magnesium - 690; manganese - 38; molyb-
d9jtum - 21; nickel -130; thallium - 20; and zinc - 48.
by rumen catheter (material given last 14 consecutive days. i.e.. day 14-
by rumen catheter (material given for 28 consecutive days)
ments where all major metabolic degra-
dation products are identified and their
retention and excretion quantitated.
However, various data summaries are
included in the project report that present
individual compound concentrations for
each animal and for each tissue or body
fluid. Some progress was also made with
the analysis of milk fat and carcass fat
samples where most of the initial
analytical difficulty was encountered. The
analytical procedures are also thoroughly
addressed in the project report.
Mammalian patterns of chlorinated hy-
drocarbon metabolism suggested that the
five compounds administered in phases I
and II, would yield several degradation
products, e.g., 3,4-dichlorophenol; 2,3-
dichlorophenol; 2,4-dichlorophenol; 2,3,5-
trichlorophenol; 2,4,5-trichlorophenol;
2,3,4.6-tetrachlorophenol; 2,3,4,5-tetra-
chlorophenol; 2,4-dichlorophenylmer-
capturic acid; 3,4-dichlorophenylmerc-
aptunc acid; pentachlorooyclohexanol;
3,4-dichlorocatechol; 4,5-dichlorocate-
chol; and tetrachlorohydroquinone.
Several of these compounds were in fact
noted in the urine samples collected
during this study. If animals were used as
biological monitors for these waste site
chemicals, the most likely indicator
compounds (i.e., compounds that indicate
previous exposure to a given group of
chemicals) would probably be the
presence of various phenols in the urine
and long-half-life lipophilic compounds in
the carcass fat.
The specific samples collected during
this project may, or may not, be
distributed as actual reference materials
depending upon user needs. However,
the analytical procedures and the
Sequence of confirming analyses (neces-
sary for sample verification) have been
established or improved during the
various phases of this project. Large
volume samples with in vivo incorporate
toxicants can be provided in subseque
efforts if additional material is needed f
actual distribution. The reference ar
quality assurance samples are intends
for use as performance evaluation mate
ials for use in selecting an analytic
laboratory from among several cand
dates or when selecting the best tecl
nique to use for a particular analysis. Th
EMSL-LV has already used the carcaj
fat and urine samples in its ongom
analytical method development an
optimization program. Many laboratorie
routinely analyze human urine sample
and the use of these reference material
should improve the accuracy of thes
determinations. Difficulties in obtamin
"safe" human blood in bulk quantities
and in finding laboratories willing to wor
with it, provides additional interest in th<
use of bovine blood for method am
laboratory performance evaluations.
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'able 3. Dosing Regime and Sample Collection Schedule for Phase III Cattle
Hereford Heifers Adult Holstein Cows
Compound
Daily dose
per animal
(grams)
Total dose
per aminal
(grams)
Daily Dose
per animal
(grams)
Total dose
per aminal
(grams)
3retreatment
Week 1
Veeki
Veek 1
Veek 1
hexachlorobenzene
yellow dye no. 3
dioctyl phthalate
ethoxyethyl acetate
hexachlorobenzene
yellow dye no. 3
dioctyl phthalate
polychlorinated btphenyls
hexachlorobenzene
yellow dye no. 3
dioctyl phthalate
methylene dianiline
polybrommated biphenyls
hexachlorobenzene
yellow dye no. 3
dioctyl phthalate
ethoxyethyl acetate
polychlorinated biphenyls
methylene dianiline
polybrommated biphenyls
carbaryl
pentachlorophenol
2.5
25
5.0
10.0
2.5
2.5
5.0
5.0
2.5
2.5
5.0
2.5
2.5
2.5
2.5
10.0
2.5
2.5
25
05
(see
final
total
below)
70
70
175
40
40
35
33
18
4
5
5
10
20
5
5
10
10
5
5
10
5
5
5
5
20
5
5
5
1
(see
final
total
below)
140
140
350
80
80
70
65
35
7
Samples Collected
Urine Milk Blood
XXX
XXX
XXX
X
XXX
'ofe: Ethoxyethyl acetate doses were discontinued after 4 days because of apparent toxicity. The PCS doses were
administered until the supply of the PCS aroclor was exhausted (8 days). The PBB doses were not administered on
the first day of week 4 due to a shortage of the PBB mixture.
able 4. Compounds Given to Phase IV Dairy Cattle.
Number of Animals Compunds Dose, Animal/Day
Total Dose/Animal
one
one
one
one
2-chlorodiphenyl ether
hexachlorobenzene
perylene
polychloronaphthalene no. 1099
i , 3,5-trichlorobenzene
diethylhexyl adipate
polychloronaphthalene no. 1014
n -dioctyl pthalate
fluorene
fluoranthene
arochlor 1260 (PCB)
tnphenyl phosphate
1 gram
1 gram
1 gram
1 gram
1 gram
1 gram
1 gram
1 gram
1 gram
1 gram
1 gram
1 gram
21 grams
21 grams
21 grams
21 grams
21 grams
21 grams
21 grams
21 grams
21 grams
21 grams •
21 grams
21 grams
Vo(ep Solids were administered orally in gelatin capsules and liquids (diethylhexyl adipate, 2-
chlorodiphenylether, and dioctyl pthalate) were infused through a rumen catheter. Blood
serum, urine, carcass fat, and liver samples were collected from the animals for ultimate use
as analytical reference materials.
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Table 5. Estimate of Analytical Detection Limits Achieved During the Current Study when Analyzing the Various
Types of Sample Material (Best Estimate Values Presented as uglg of Sample Material)
Detection Limits (uglg)
Achieved Using Different Sample Materials
Compound
tetrachlorobenzene
carbaryl
methylene dianiline
dichlorophenols
trichlorophenols
tetrachlorobenzene
dioctyl phthalate
lindane
yellow dye no. 3
polychlorinated biphenyls
polybrommated biphenyls
ethoxyethyl acetate
pentachlorophenol
dich/orobenzne
hexachlorobenzene
Blood
Serum
0.01
0.02
0.02
0.01
0.01
0.01
0.01
0.01
0.02
0.02
0.05
0.01
0.02
0.01
0.5
Milk
0.01
NO
ND
0.01
0.01
0.01
ND
0.01
ND
ND
ND
ND
0.01
0.01
ND
Milk Fat
ND
0.05
0.5
ND
ND
ND
0.1
ND
1.0
0.02
0.05
ND
0.2
ND
0.01
Urine
0.001
0.01
0.01
0.01
0.01
0.001
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.001
0.01
Feces
0.01
ND
ND
0.01
0.03
0.01
ND
0.05
ND
ND
ND
ND
0.05
0.01
ND
Liver
0.01
0.1
ND
0.01
0.03
0.01
0.20
0.03
1.0
0.5
0.5
ND
1.0
0.01
0.1
Kidney
0.01
No
ND
0.01
0.02
0.01
ND
0.03
ND
ND
ND
ND
0.03
0.01
ND
Carcass
Fat
0.03
0.1
ND
0.03
0.1
0.03
02
0.01
1.0
0.5
0.5
ND
1.0
0.03
1.0
Note: Varying amounts of analytical interference were typically encountered from sample to sample and the above
values are presented as an approximation of the detection limits actually achieved when analyzing the different
types of sample material. Carcass fat, milk, and milk fat were probably the most difficult of the samples to
analyze. The notation ND (not determined) means that a detection limit estimate was not achieved when using
the indicated sample material.
Table 6. Determination of Analytical Precision Based on Analysis of 10 Separate Aliquots of a
Single Urine Sample. (Precision Expressed as Coefficient of Variation for 11 in vivo
Incorporated Compounds Present in the Urine.)
Compound
tetrachlorobenzene
tetrachlorophenol
2,4,5-trichlorophenol
1,2-dichlorobenzene
3,4-dichlorophenol
2,3- and 2,4-dichlorophenol
lindane
phenol
methyl phenol
tetrachlorohydroqumone
pentachlorophenol
Number of
Determinations
10
10
10
10
10
10
10
10
10
10
10
Mean
Concentration
(pg'g)
0.05
27.78
5.74
0.13
7.07
8.27
0.19
3.39
7.65
2.42
42.70
Coefficient of
Variation (%)
40
35
38
15
44
33
37
44
56
44
35
Note: Ten aliquots of a single urine sample were taken and sequentially analyzed. Sample preparation
steps, GC/MS analysis, and routine quality control steps were conducted for each aliquot.
-------
-------
V. R. Bohman, C. R. Blincoe, G. C. Miller, and R. L. Scholl are with the University
of Nevada, Reno, NV 89557; the EPA authors, W. W. Sutton (also the EPA Project
Officer, see below) and L R. Williams, are with the Environmental Monitoring
Systems Laboratory, Las Vegas, NV 89193-3478.
The complete report, entitled "Biological Monitoring Systems for Hazardous Waste
Sites (Production and Analysis of Analytical Reference Materials)," (Order No. PB
89-t*9204/AS; Cost: $21.95, subject to change) will be available only from:
National Technical Information Service
5285 Port Royal Road
Springfield, VA 22161
Telephone: 703-487-4650
The EPA Project Officer can be contacted at:
Environmental Monitoring Systems Laboratory
U.S. Environmental Protection Agency
Las Vegas, NV 89193-3478
United States
Environmental Protection
Agency
Center for Environmental Research
Information
Cincinnati OH 45268
I
' Official Business
, Penalty for Private Use $300
EPA/600/S4-89/007
CC0085833 PS
'SE'CI
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