TD480
.525
1977
PROCEEDINGS:
SEMINAR ON ANALYTICAL METHODS
FOR PRIORITY POLLUTANTS
°°OR7710.
RECEIVED
FE8 29 1980
AGENCY
LIBRARY, REGION V
Environmental Protection Agency
November 9 & 10, 1977
Denver, Colorado
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UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
OATE: i ? MAR 1978
SUBJECT: Proceedings-Seminar on Analytical Methods for Priority Pollutants
William A. Telliard, Chief
Energy & Mining Branch
TOT Robert B. Schaffer, Director
Effluent Guidelines Division
On November 9 & 10, 1977, a seminar was conducted, in Denver, Colorado,
on the subject of Analytical Methods for Priority Pollutants. Nearly
one hundred people attended this seminar, among them E.P.A. staff,
industrial trade association representatives and technical contractors
for this division. A complete list of attendees is included herein.
From the seminar a record was prepared, which is presented with this
memorandum.
The purpose of this package is to provide those concerned and working
with Sampling and Analysis Procedures forthe Screening of Industrial
Eff1uents for Priority Pollutants (the protocol), an account of the
issues discussed. These notes are organized following the same order as
the agenda for the seminar. All topics of discussion have been restruc-
tured into a series of issues, with their associated discussion and
resolutions. Furthermore, any comments or references submitted in writing
are also included. In preparing this total package, it was considered
advantageous to take the time to accept written comments for the record,
as well as prepare a summary and table showing in which industries
priority pollutants were found.
In addition to presenting a record of the seminar, an effort was made to
recommend or suggest alternative ways of resolving any issues on the
use of the protocol. It appears evident that those people attending this
seminar found it to be useful. An opportunity was given to people using
the protocol, to bring their questions and suggestions to a group of
experienced chemists for consideration. In this regard, we have managed
to straighten out and refine the analytical methods being used for
BAT review studies. Moreover it seems evident that the protocol is a work-
able, reliable manual. Many laboratories are working with these procedures
and have commented that they are effective in meeting the needs of the
screening phase studies. The overall accomplishment of the seminar was
one of fine-tuning the protocol.
EPA FORM 1320-6 IRSV. 1-761
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Table of Content
Page
I. Proceedings 1
II. Coimients 80
III. References 156
IV. Attendees 261
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Seminar on Analytical Methods
for Priority Pollutants
Agenda
November 9, 1977
I. Introduction
II. Organic Analysis
A. Screening Phase
1. Purge and Trap - GC-MS
B.
Discussion Leaders
W. A. Telliard
Tom Bellar, Jim Lichtenberg, Clarence Haile,
and Jim Spigarelli
2. Liquid - Liquid Extraction- Walter Shackelford and Paul Taylor
GC-MS
Verification Phase
Methods validation, Quality
Control and Documentation Data
Walter Shackelford, Jim Lichtenberg, and
Ron Kagel
November 10. 1977
III.Metals Analysis
IV. Asbestos Measurement
Billy Fairless and Mark Carter
Charles Anderson, Martha Bronstein,
Michael Terlecky and Phil Cook
V. Biological Monitoring
Charles Stephan and Gary Raw!ings
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VOA - Purge and Trap GC/MS
Introduction
The first day of the seminar was dedicated to the review of the
procedures being employed for organic analysis. The morning session
was dedicated to the review of the volatile organic analysis (VOA) and
any issues or questions revolving about this particular aspect of the
program. During the discussions such subjects as sampling, storage,
compositing and the use of internal standards were addressed. In
addition, questions regarding operating conditions and alternative
procedures were also presented by the various participants. Since the
close of the meeting, a number of written comments have been received
in addition to the oral ones presented at the meeting and these
comments will be noted throughout the proceedings.
Issue: Sample collection. Sample Site and Number of Samples
Discussion:
A number of questions were raised regarding the VOA samples with
respect to collection, spiking, number of samples and container size.
Standard Oil; When should you spike a VOA sample in the field?
EPA: Spike a sample only before capping it. Do not pierce the septum
with a needle.
Shell Comments: A number of questions were raised including:
1. what is the maximum storage time for VOA's at 4°C (without
formation of bubbles)?
2. how many vials must be sampled?
3. can we composite VOA samples?
4. what is optimum volume for a sample vial?
Shell recommends a 45 ml vial; 125 ml is too large. See a more
complete version of Shell comments in the comments section.
Resolution:
The protocol specifies a minimum of one sample per 24 hour period to
be collected in duplicate. This is a minimum requirement and there is
no reason why more samples could not be taken if sample crews were on
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site. EPA, Cincinnati/ recommends the use of teflon faced septum,
sealed screw cap bottles. Glass vials with Bakelite screw caps with a
hole in the center, as described in the protocol, have proven superior
to crimp cap serum bottles.
Issue: Sampling and Preservation of Chlorinated Effluents or VGA
Analysis
Discussion:
RETA has indicated in written comments (see the comment section) that
the potassium iodide indicator paper specified in the protocol, was
not adequately meeting the requirements of the field crews. Moreover
they have gone to chlorometric procedures for determining residual
chlorine.
A question was raised as to the affects of preserving with sodium
thiosulfate. Reference was made to a paper prepared by C. Carol
Morris of Havard which discussed such a problem.
Status:
At the present time, for chlorinated effluents, the field crews are to
sample for two sets preserving one and not preserving the other. We
would expect the contractor to run both the preserved and the
unpreserved samples until such time as additional data can be gathered
as to the total effect or lack there of, of the preservation of the
free chlorine.
Issue: suggested alternative, internal standards for use in VGA
analysis.
Discussion:
Midwest Research replied to questions.
Q Do you have a problem with use of D-chloroform as a standard? Are
there any cross contributions?
A. No problems.
Q. What level of concentration of D-chloroform was used?
A. 10 ppb.
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Q. Did you find chloroform on the chromatogram while using
D-chloroform as a standard?
A. Yes.
Midwest discussed problems with use of internal standards for
volatiles. Protocol specifies use of bromochloromethane,
2-bromo-1-chloropropane and 1,4-dichlorobutane. They found
interferences with these: Recommended use of D-chloroform and
D8-toluene, found no interferences, generally good performers. D<3
toluene was found to be cheap, available and not generally found
in industrial effluents.
Midwest suggests use of multiple standards.
Monsanto Comments on Internal Standards
Monsanto has been measuring priority pollutants for the textile
industry.
Problems included:
locating a source of 2-bromo-1-chloropropane;
use of 1,U-dichlorobutane; this compound elutes close to toluene; need
an internal standard that elutes independently;
How pure is D-toluene?
Monsanto is concerned because toluene is seen in many effluents from
this industry.
Recommended Status:
EPA still strongly recommends following the Protocol. If a laboratory
finds that it is necessary to use an additional standard, this will be
acceptable as long as it is adequately documented.
Issue: Internal GC/MS standards for the very volatile fraction
Discussion:
Radian: Noted that they have a problem storing very volatile
standards, particularly vinyl chloride. Currently they are preparing
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new standards in Teflon sealed hypovials under an argon atmosphere to
prevent loss of the standard. However, they still have problems with
volatile components.
EPA (Athens); Suggestion, make your own standards for very volatile
compounds in the lab.
MRI: Stores sample tubes of vinyl chloride at 4°C and adds a plug of
fresh adsorbent to minimize losses.
Resolution:
It is recommended that individual laboratories prepare their own
standards for such things as vinyl chloride, methyl chloride,
methylbromide, chloroethane and dichlorofluoromethane.
Issue: storage of VGA samples prior to analysis
Discussion:
Q. (RETA) What is the validity of sample results after purged samples
have been stored in traps?
A. (EPA Cincinnati) The olefins rearrange at U°C to cis-trans
isomeric forms. Compounds may also migrate on the trap, resulting
in a change in peak geometry. In general, storing traps is not
recommended. There are still too many unanswered questions.
NUS Comments: They do not recommend the storage of trap samples.
They get unknowns which they cannot identify. Do not freeze trap
samples. (See NUS findings in the reference section).
EPA (Kansas City): Reports they have stored VOA samples for up to 2
weeks. They place sealed vials into dessicators with activated carbon
to prevent contamination with methylene chloride.
NUS Comments: They seal VOA trapped samples in metal cans and freeze
them; or they store traps in glass tubes; seal ends of traps with
Teflon and stainless steel. Then check seals after cooling to prevent
loosening at Teflon caps.
RETA: They seal tubes with heat and store them under helium
atmosphere and refrigeration. Storage results in wierd peaks;
however, there were no losses. They accidentally analyzed a clean
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trap that had been stored for 2 weeks and obtained the same wierd
peaks. Could packing be the source?
Jacobs to EPA; Problems concerning the storage of samples must be
resolved.
EPA (Cincinnati) ; Samples should be stored in vials, not on purge
traps. Vials can be stored 2 weeks at 4°C with no noted loss of
compounds.
Shell: Stored vials for a period of 2 weeks at 4°C with no problems;
however, 21 day storage resulted in a reduction of compounds.
RETA: States that they stored some samples as long as two months.
Recommendation: The VOA samples should be preserved in teflon sealed
containers at 4°C and in darkness. It is recommended, from the data
and information available, that the samples be held no longer than two
weeks prior to analysis and that the samples should not be transferred
to traps for storage but rather left in their own containers.
Issue: Contamination of VOA samples with methylene chloride
Discussion:
A number of methods were recommended to protect the VOA samples prior
to analysis from contamination. EPA Region VII laboratory suggested
that the samples be stored in a desiccator containing activated
carbon.
Monsanto: Comments that tubes trapped and sealed under a nitrogen
atmosphere prevents contamination with methylene chloride. Monsanto
further suggests that methanol used to spike the original sample may
be a source of methylene chloride contamination.
Shell suggests prestoring of VOA samples in the field in a pint jar.
This procedure prevents contamination with methylene chloride.
Resolution:
This suggestion from EPA Region VII would be useful if the problem is
due to the laboratories layout or configuration. Then it is advisable
to insure it against contamination by methylene chloride. The
suggestion by Shell is also very useful.
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Issue: Compositing VGA samples
Discussion:
Shell questioned whether, under the screening, it was permissable to
composite the VGA samples. In addition. Shell has provided data (see
their full comments) on the applicability of compositing the sample in
the trap from a number of VGA samples.
Shell* s comments on compositing included:
1. Their technique takes 3 individual grab samples during a 24-hour
period and composites these samples in the Tekmar unit by
injecting 2 ml from each grab sample.
2. One individual grab sample is too small to represent an entire
waste stream.
OCC; Requested information on compositing VGA samples.
EPA (Cincinnati) ; Suggests pouring 2 ml from each of 3 samples into
syringe. EPA prefers lab compositing, since field compositing tends
to result in loss of low boilers.
NUS: Found that pouring VGA samples into a syringe resulted in a loss
of methylene chloride.
Resolution:
Compositing of VGA's has been an acceptable practice and will continue
to be so. The selection of syringe versus open pouring technique will
be left to the analyst but must be documented.
Issue: Some considerations relating to the use of silica gel in the
purge and trap technique.
Discussion:
Several comments were concerned with the collection and buildup of
water in the tenax trap. They requested some alternatives or
techniques to minimize the problem.
Midwest Research Institute (MRI) found problems with very volatile
compounds, i.e., chloromethane, dichlorofluoromethane, vinyl chloride.
These problems included water retention with use of silica gel and
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bleed of column packing into GC/MS; water dumping on MS was also
noted. Their solution: proceed without the use of silica gel; no
problems encountered. Carborsive was suggested as an alternative to
silica gel.
EPA Cincinnati commented: change silica gel often (1-2 times/week) to
eliminate water retention.
Resolution:
EPA suggests that the operators simply increase the number of times
per week that the silica gel is changed to insure no water buildup.
Issue: Mechanical and operating problems with the Tekmer Unit.
Discussion:
A number of commenters presented data related to operating problems as
a result of certain mechanical or physical faults within the Tekmar
unit. The following comments presented by Tom Bellar set forth the
basic information. The second commenter, Shell presented a lengthy
description of its reprogramming of its Tekmar. The new heap tracing
diagram showing changes that Shell had made in its particular unit is
presented in their full written comments.
Comments by EPA Cincinnati on Tekmar GCMS:
have tested units 1, 2 and 3 and have uncovered a variety of
problems with these units
1. trap heater does not heat quickly enough; should be able to get
180°C in less than 1 min.; suggests use of heating tape to wrap
trap in order to maintain temperature
2. trap may move inside oven - can be problem; make sure end of trap
is heated sufficiently to give desorption.
3. check teflon plugs that hold desorbent in place; make sure it's
secure.
4. temperature sensor may not monitor actual trap temperature; make
sure trap is cooled to room temperature before next run.
Gave specifications of packing procedure
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trap should be 24 cm long or longer
ID should be 0.105 in
wall thickness = 0.01 in
material should be 304 SS seamless
Packing:
5 mm glass wool plug
1 cm 3% OV-1 chromosorb 60/80 mesh
16 cm Tenax CG 60/80 mesh
8 cm silica gel - grade 15
5 mm glass wool plug
Recommended conditioning overnight with 20 ml/min purge gas, 200°C to
ensure entire trap is heated; make sure all tubing is heated to remove
water. Necessary to put purge gas filter in system; a1/4 Ib 13x
molecular sieve removes many interferences. Recommends frequent
changing to reduce interference,
Shell's problems with Tekmar unit included;
1. Need to head trace every line including switching valve to
eliminate water.
2. Teflon valves on ends of tubes need replacement
3. Samplers must-be cleaned and stored at 105° to remove memory
effects,
4. Need to replace brace fittings and o-rings with stainless steel
fittings with Teflon barrels to eliminate memory effects. Shell
operates sampler at 70°C to get better desorption and reduce
memory effect. They also heat the transfer lines for the same
reasons.
EPA (Cincinnati): Does not heat purging device because heating gives
them increased chloroform values. In order to resolve early eluting
compounds, the entire trap must be heatad to 180°C in 30 seconds.
Bellar1 s equipment takes 15 seconds.
Issue: Direct Aqueous Injection
Comments were received regarding acrolein and acrylonitrile in the
direct aqueous method.
Discussion:
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Region VII in their written comments have presented an argument that
the present procedure is both costly in time and manpower and
recommend that we look at an elevated temperature for the purging
technique to encompass all of the volatile compounds.
FMC: Uses direct aqueous injection and injects 10-20 ul samples into
a Tenax column.
MRI: Requested information about direct aqueous injection techniques.
EPA: Further investigations on this technique are proceeding.
Issue: Recovery data for the VGA analysis
Discussion:
Region VII S & A laboratory in their written comments have supplied
some limited recovery data on analysis of VGA which they performed.
This data is presented in the comment section.
DuPont has also submitted written comments to the Agency regarding the
VGA procedure. Specific information contained under the comment
section under DuPont would merit your review. Some comparative data
is provided showing a comparison of the analysis performed by the
steam electric industries contractor, NUS and the Agency's contractor,
California Analytical Laboratories. The data presented shows a
comparison between samples which were split between Mobil Chemical and
the Agency and a comparison of both their GC/MS as well as their GC/UV
are presented in these tables. Metals and classic parameter analysis
are also compared.
Liquid-Liquid Extraction
EPA (Athens): In the course of screening analysis, three liquid-liquid
extractions are performed (the acid, base/neutral, and pesticide
fractions). The following questions refer to these extractions:
1. Are continuous extraction techniques being used?
2. Are contractors extracting two liters of sample?
3. What methods are being used to break emulsions?
4. Is 85 percent of the solvent used in the extraction being
recovered?
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Continuous Extractor
Midwest Research Institute (MRI): We have used continuous
liquid-liquid extractors for the tanning industry. In general we have
found extractions difficult due to emulsions. See diagram of
apparatus in the reference section.
FMC: We have had problems with efficiency of base neutral extractions
using a continuous extractor. In some cases we have had to spend as
long as 24 hours in extraction using 50 milliliters of solvent.
California Analytical Laboratories (CAL): Use of less solvent to
avoid dilution effects as well as an increased reflux rate can help
your extraction problem.
EPA (Athens): We have been using a Hershberg-Wolfe continuous
extractor available from Ace Glass, part number 6841-10. The
advantage of this apparatus is reproducible droplet size. Only one
liter of sample may be used, but this is no problem since the samples
that form emulsions are generally too concentrated for the two liter
sample to be used.
Analytical Research Laboratories, Inc. (ARLI): We have designed our
own three stage extractor and gotten good recoveries. It has run as
long as 3-4 days. It works well with marine waters but some problems
with sediments are evident.
Hydroscience: We have used the Aldritch continuous extractor but plan
to use shake-out due to the difficulty in cleaning the continuous
extractor.
EPA (Athens): The muffle oven at 400°C may be used for cleaning.
With the 4-liter extractor the solvent rinse and drying techniques
must be used due to the size of the extractor.
National Council: We have not had success with continuous extraction
on pulp mill samples.
Recommendation:
In the protocol, it is written that a continuous liquid-liquid
extractor should be used when emulsions form. No specific type of
apparatus is specified. A laboratory should document what they used.
Extraction Volume
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CAL: We routinely use 1700 nu.llili.ters of sample for shake-out
extractions with 250 milliliters of methylene chloride solvent. We
continue to have problems with paint and ink process samples. These
may be candidates for continuous extraction.
Emulsion Breaking and Solvent Recovery
DuPont: We use centrifugation for breaking emulsions.
EPA (Cincinnati): We have used centrifugation, also. 500 milliliter
bottles are centrifuged at 1000 RPM for 5 minutes.
Monsanto (Dayton): We use centrifugation at 4,000 rpm for 4 minutes
and are able to separate emulsions on textile samples. We recover 85
percent of the solvent.
Hydroscience: We generally do not achieve 85 percent recovery of
solvent but find that recovery ranges from 50 percent to 90 percent
depending on sample character. Some samples seem to pick up solvent.
CAL: More solvent may be used to increase solvent recovery.
EPA (Region VII) : We normally achieve 85 percent solvent recovery.
Our laboratory uses filtration through glass wool packs for breaking
emulsions.
Standards
EPA (Athens): Some users of the Radian standards have complained that
diphenylhydrazine is being converted to azobenzene. Studies at the
Athens lab indicate that 1, 2-diphenylhydrazine decomposes to
azobenzene in a number of solvents including methylene chloride and
water. N-nitroso diphenylamine appears to decompose in the GC
injection port at temperatures greater than 100°.
Monsanto (Dayton): We have experienced disappearance of d-10
anthracene in 10 percent of our samples and feel that anthracene may
react in the procedure. We suggest spiking with 3 internal standards,
D-anthracene and 2 others.
Acid Extract
Environmental Science and Engineering (ESSE) : We have been analyzing
phenols present in process waters from the wood preserving industry.
It appears to us that the extraction procedure outlined in the EPA
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protocol does not extract phenols efficiently. At present we are
using the steam distillation method outlined in Analytical Chemistry,
1975, 47, 1325-29. We have data comparing the steam distillation to
the shakeout method of extraction. (See E S & E written comments in
the reference section).
EPA (Region VII) : From our work we feel that the preservation methods
may be masking phenols.
EPA (Athens): These samples are not preserved. Our work on similar
samples to ES&E indicates that there are no problems with loss of
phenols from degradation at high pH. In samples that form emulsions,
however, it appears that the extraction efficiency of phenols drops
significantly. Dse of the continuous extractor in these cases
improves extraction efficiency.
EPA (Region V): We have found that phenols can be preserved at pH >
11 if kept cold.
EPA (Region VII): Why is a grab sample for phenols taken as well as
the acid fraction of the composite sample.
EPA: The grab sample is taken strictly for the classical 4AAP analysis
for total phenols. The extraction followed by GC/MS is for the
determination of 11 specific phenolic compounds.
EPA (Region IV) : We have tried an experimental packing from Supelco
for phenolics. It appears to work well but degrades quickly. We are
evaluating this packing further.
EPA (Athens) : Whereas Tenax GC has some faults, it nevertheless elutes
all 11 of the phenolics.
EPA (Region V) : Could not the phenols be derivatized? We realize
that this could create more problems.
EPA (Athens): We have tried derivitization. It seems that while
diazomethane derivitization works well, the extracts cannot be stored.
Quenching of the reaction with acetic acid did not stop the
degradation of components in the extract. Pentafluorobenzyl
derivatives were also made but these were difficult to synthesize.
Mobil Research: We find interferences from isomers of dimethylphenol
in our analysis.
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MRI: Capillary columns may separate the isomers in question.
Conclusion:
Based on work done by this Agency and some of our contractors, we find
that the assertion that phenols are destroyed at a high pH is not
justified. Moreover, the use of steam distillation for a clean-up was
discussed and accepted earlier. Under the present program
derivatization is not an acceptable alternative.
Concentration and Extraction Handling
EPA (Athens): It has been pointed out that laboratories that are new
to Kuderna-Danish evaporation of solvent may try to heat the solution
slowly to avoid loss of components. In fact the reverse is true.
Only by quick heating of the solvent can good recoveries of components
be obtained.
EPA (Athens): The drying of extracts with sodium sulfate is a
controversial subject. We feel that water is driven from the extract
during K-D evaporation, as an azeotrope.
Shell Development: We believe that the drying step should be
eliminated since it is a possible source of contamination. Also, 50
percent of the organics may be adsorbed by the sodium sulfate. We
question the benefits of drying.
EPA (Cincinnati): We feel that drying the extract with sodium sulfate
is a necessary step in processing the extract. Previous data
documents this step and no data has been presented for effluent
extracts without drying. The drying step also aids in separation of
phases when emulsions are formed.
MRI: To avoid contamination of the extract by organics in the sodium
sulfate -we ash it at 650° in a muffle furnace and rinse with hexane.
Mobil Research: Where can sodium sulfate be obtained of a quality
necessary for this procedure? How does one insure it is clean?
EPA (Cincinnati): We use Mallinkrodt granular. If an artifact
persists, heat a shallow dish of the sodium sulfate at 400° for 2-3
hours.
DuPont; We use an additional blank of solvent through the drying
tube.
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Shell Development: We find that the drying takes 1-1.5 hours and
costs too much in time to justify the supposed benefits.
EPA (Cincinnati): We do not take nearly this much time. How large is
your drying tube?
Shell Development: 8 mm in diameter.
EPA (Cincinnati): Our tubes are 19-20 mm in diameter. Perhaps this
is your problem.
MRI: Too much water in the drying tube can cause plugging. This
could be checked, too.
EPA (Region IV): We have used a glass wool filter to prevent solids
and water from plugging the drying tube.
Conclusion:
Extract drying using sodium sulfate remains the preferred procedure
for residual water clean-up. Quality reagent and care in its use will
prevent an introduction of contaminants through the drying step.
Polynuclear Aromatic Hydrocarbons
Mobil Research: We are seeing two problems in PAH identification.
First, we cannot separate benz(a)anthracene and chrysene on our GC so
we must report the peak as a combination of the two. Also, we find
that in our effluent perylene interferes with benz(a)pyrene giving
erroneously high results for benz(a)pyrene.
EPA (Athens): The retention times given for benz(a)anthracene and
chrysene in the protocol are erroneous. These two cannot be separated
on the recommended column. Our contractors are reporting these as the
sum of the two. We were not aware of the perylene interferences with
benz(a)pyrene. For the verification stage of analysis this must be
taken into account. We are aware of your lab's work with GC-OV as a
determining method.
Verification, Methods Validation and Quality Control
Manufacturing Chemist Association (MCA): We have a variety of
concerns about the verification program. Before proceeding (from
screening analysis to verification) three things should be
established: (for these written comments, see the reference section).
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(1) Define the analytical methods for the priority pollutants;
what constitutes a limit of detection; in what manner is the
data to be reported?
(2) The protocol needs definition: things such as the complex
nature of industrial effluents; the VOA technique and the
asbestos technique need clarification.
(3) Criteria should be specified for a given method so that
alternate methods meeting those criteria could be used.
(4) Detection limits for the instrument have to be specified:
(a) the signal/noise ratio should be 2.5 to 1 or reported as
not detected.
(b) sample should be rerun if the signal to noise ratio is
low.
(5) Industry will challenge any and all data if the protocol is
not specified more clearly.
EPA: The protocol was designed for screening analysis only.
Verification methods have been under study for some time.
EPA (Cincinnati) : Our policy for validation is as follows:
(1) The method must be an established method.
(2) Concentration levels should reflect those expected to be
present. Thus, a variety of concentration ranges in
distilled water and in the sample type should be studied
(3) Dosed distilled water samples for the ideal case should be
analyzed in round robin fashion.
Dosed field samples should be done in round robin.
Round robin parameters should be:
(a) 75 to 100 labs (ideally)
(b) minimum of 15 labs should return usable data
(c) minimum of 3 concentration levels should be used
(d) comparison of distilled water & sample data should be made
(e) outliers in data results should be rejected
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EPA (EGD): We suggest 3 labs analyzing one sample. There is a need
to agree upon criteria (i.e. method steps) for any analytical
technique.
Mobil Research: Final validation should be done on actual (typical)
waste water samples both a high and low concentration.
EPA: One possible validation scheme could be:
3 effluents
3 ranges
3 labs
3 replicates
American Cyanamid: Why are verification programs proceeding without
an agreement between industry and the EPA on what constitutes a
verification method/program?
EPA (EGD): The court dates (deadlines) remain whether or not we can
agree on methods. Sampling for the verification must continue.
NUS: How will the verification program be altered after validation
methods are established?
EPA: Changes (if any) will vary from project to project. It is
possible that there will have to be revisits to the field for
additional sampling.
NUS: Based on the deadline dates, validation for the methods will
come after verification sampling has ended. In any event, the issue
will probably wind up being solved in court.
EPA (EGD): What are your thoughts on the 3 lab, 3 sample, 3
concentration and 3 replicate validation?
EPA (Denver): We should look at more sample types and fewer
concentration levels. Matrix interference is our outstanding problem.
EPA (Region V): A group of people will be gathered to discuss a
validation scheme. The scheme should cover all compounds on protocol
not just those found in the screening effort.
End of Day 1
Day 2 - Methods Validation
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In a discussion the previous night a scheme for method validation was
discussed and held to have merit by EPA and industrial chemists. The
parameters were:
(a) 3 labs
(b) 7 determinations
(c) 7 spikes (3 concentrations)
(d) 7 samples
(e) validation done for only those compounds found in screening.
CAL: How can EPA arrive at a standard method which does not validate
those compounds which are not found in the screening effort?
EPA (Region V) : We also feel that all compounds should have validated
methods.
EPA (EGD): There is not enough time or money to support a research
program to validate methods for all compounds. Court dates have to be
met.
Catalytic: What is the possibility of validating the screening
analysis? We are concerned about the stability and storage of samples
from field to lot.
EPA: There is no easy way to validate field screening procedures in
the time left to do it.
Radian: Could the contractors spike some samples and analyze later?
EPA: This is a possibility as well as having a referee laboratory.
National Council: Many of the industrial effluents are heavy in
solids and compound associated solids. Can one demonstrate by spiking
the sample what is actually there? The sample, after all, is an
extract from the real environment.
MRI: There are two consideration in view of the limited time
available:
(a) Quality control on all steps of the procedure.
(b) Extensive validation on a few steps but many samples run.
Recommendation:
18
-------
This issue is unresolved. EPA at Cincinnati is considering the
problem in an attempt to find a practical solution.
Use of Blanks
There has been some confusion concerning the number of blanks
necessary in the screening phase. Let's discuss the blanks for the
volatile organic analysis (VGA). Each VOA sample should be collected
in duplicate. This is not to say that each of these samples must be
run separately. They simply provide, to the analyst, a backup sample
if for some reason he should have difficulty with the first sample.
Once a sample is opened, it has no further value, hence the need for
duplicates. There should be a blank VOA sample which would be
prebottled in the laboratory, taken to the facility, carried through
the procedures and exposed to the various conditions at the facility
during the sampling run. Therefore, for each plant there should be
one VOA sample blank not a VOA sample blank for each point. The
second issue is that of the use of field blanks for compositing
samplers. The purpose of the field blank is to insure that
contamination is not being picked up either from an inadequately
cleaned sampler or a contaminated intake line. A number of options
are provided for the use of field blanks, one, is of course, the use
of manual compositing which would do away with the need for any field
blank another option is instead of running the organic-free water
through the sampler, the individual may utilize the water supply for
that plant, i.e. city water, well water, river water, what ever, and
that may be run through the sampler to purge it prior to use. At
least in this particular case the need for one additional analysis is
eliminated. The third option, which leaves much to be desired, would
be the compositing of all the field blanks prior to analysis. This
would lend very little creedance and require some additional sampling
if you are looking for total background. The use of a sampler or
field blank in sampling in process lines probably has limited
application. That is to say, due to the high concentration and heavy
loadings of these particular lines, the minor contamination that might
be present probably wouldn't be noticed and probably wouldn't be
looked for. Therefore as far as verification is concerned the
application of influent field blanks probably lends little if any
assistance to the program.
Data Reporting
Enclosed are two sets of data reporting formats. The first is the
format to be utilized in reporting GC/MS screening analysis by our
contractors and our regional offices. We would hope that both the
19
-------
Environmental Research Laboratory
UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
Athens, Georgia 30605
DATE: December 28, 1977
SUBJECT: Format For Storage of Mass Spectrometry Data on 9-Track
Magnetic Tape Proposed by Carborundum
FROM; w> M> shackelford
Analytical Chemistry Branch
T0: William A. Telliard
Environmental Protection Agency
Effluent Guidelines Division
WH-552, 401 M Street, SW
Washington, DC 20460
The Carborundum Company has proposed a format for saving mass
spectrometry data on 9-track magnetic tape. I have received
a sample tape as well as documentation, and our computer
center has read the tape and even written a simple program
to plot out the data. I am sending along a copy of the
documentation as well as a chromatogram reconstructed from
the sample data.
Since this format was developed for Carborundum by Finnigan,
we can expect all contractors who utilize the Finnigan-Incos
GC-MS-computer system to be able to use this format.
I recommend that the proposed format be accepted. Bob
Fluege of Carborundum is awaiting notification by the
project officer.
Attachment
20
EPA FORM 1320-6 !REV. 3-7S)
-------
EXfiMEPA. DS
THIS FILE DOCUMENTS THE FORMAT OF .EP FILES WRITTEN BY THE
MS US COMMAND 'EPfi' .
WHAT FOLLOWS IS THE CONTENTS OF THE FILE E031.EP AFTER THE
MSDS COMMAND 'EPA BOS1, , BOS1/D' IS EXECUTED.
SEE 'EXPLEPA.DS' FOR A DISCUSSION OF THE ARGUMENTS.
(THE /D MODIFIER WAS USED TO MAKE THIS OUTPUT PRETTY
BY PUTTING A 'CR' AFTER EACH 83 CHARACTER LOGIAL RECORD.)
• THE SIX QUESTIONS ASKED BY 'EPA' WERE AN3UERED AS FOLLOWS:
FIRST SCAN TO SAVE <1>, 4
LAST SCAN TO SCAN (268): 6
LOWEST MASS TO CAVE C28>: 'CR'
HIGHEST MASS TO SAVE (250): 'CR'
MINIMUM INTENSITY TO SAVE IN IONS <1>: 'CR'
MINIMUM INTENSITY TO SAVE AS "'. BASE <. 1 > = 'CR'
(THE ANSWER 'CR' MEANS USE THE PROMPT VALUE.
THE PROMPT SCAN LIMITS ARE THE LIMITS OF THE DATA.
THE PROMPT MASS LIMITS ARE THE SCANNED REGION.
THE PROMPT MINIMUM INTENSIFIES HfiE THE SMALLEST ALLOWED.
LARGER VALUES CAN RESULT IN SUBSTANTIAL REDUCTIONS IN
• THE DISK SPACE AND-CPU TIME USED.)
BOS1 # 8 07/03x75 13=58
HC STANDARD INST = C
SECS/SCAN:
ANAL: JEC SUB: JEC ACT: NONE FOR: K M: 2
BOS1 # 4 07/89/75 13=50:08 + 0=24 BASE 63, 3663. RIC
027834323033338813831064858622051890653832862887665084863993676812375683331
082088033004835005033025835007186843181812112984113828117882113224128885124
131132132803133010143016144001145003147003150003151821155083162319153913163
170003174w*2175835181170182887183386136085137305193023194882195802201086285
207801212Q8721308721788721984322400322508523103423208523380223(50 18243631244
090888
BOS1 ft 5 67/83x75 13=58:88 + 3=38 BASE 69. 3793. RIC
02703803881703186583638204468464503365332 435133486283:786380o36993987380903i
032002083806085883837801333021835805103844181883112083113930119232120e06124
1311 S3 132Q871333111358^2137 802143815i 45853158387151825155003162815163912163
170884175882131172132609133386•96386193825134082233082281903285813212607213
217066219062225002229862231033232003233063236087243047244002245001243002006
BOS1 # 6 07/03/75 13=50=36 + 8=36 BASE £9, 19,13. RIC
02787383tt825831835044088858019651398862804065006069393075003031635032005033
08508905608483382389563518804718101611283611303711$279131224132003133805143
1450351506831510241550111620301630211^9193178004175803131250132914133387136
193033195063290064281864285835212015213811217036219189220835225034231153232
236003237805243873888888
C WHAT FOLLOWS DESCRIBES TH£ FORMAT OF THE PRECEDING DATA.
C THERE IS A 4 LINE ; (ASSUMING 2 CHARACTERS/WORD)
R£A:»-:DSK. no; H/^'i?:, ISCAN, IDATE, IHOUR, IMIN
110 -TjRMAT (&H2, IX, Ib, 2V-., 4A2, 13, IX, I2>
-------
DATA READ:
C '
c
C
C'
c
NAME
ISCfiH
I DATE
I HOUR
I M I H
129
C
C
C
13Q
C
C
c
140
C
C
C
c
c
c
c
c
c
c
c
c
c
c
;12 CHARACTER NAME OF ORIGINAL DATA FILE
;0 TO FLAG FILE HEADER FIRST LINE
;8 CHARACTER DATE A3 'MM/DD'YY'
;HOUR AT START OF RUN
;MINUTE AT START OF RUN
THE SECOND HEftDER LINE CAN BE READ AS FOLLOWS:
INTEGER I SAMP-; 32 ), INST<3>
READ
R£AD
-------
FORMATU3U3, 13), !2>
DATA READ-.
MASSU) JNOMINAL MASS (STRICTLY INCREASING)
;MAS3
-------
IN I tNSI TY
24
CO
Xi
rn
n
o
oo
o
CO
CD
-------
SAWfL
DATE
STD.
CONC.
Date
NO.1
IB
2V
3V
4V
5B
6V
7V
8B
9B
10V
11 V
12B
13V
14V
15V
16V
17B
18B
15V
20B
21A
22A
23V
24A
25B
26B
27B
28B
29V
30V
t i.U.
INJECTED
I.D.
FACTOR
Extracted
COMPOUND
acenaphthene
acrolein
acrylonitrile
benzene
benzidine
carbon tetrachloride
chlorobenzene
1,2,3, -trichlorobenzene
hexachlorobenzene
1,2-dichloroethane
1 , 1 , 1-trichloroethane
hexachloroethane
1,1-dichlo roe thane
1,1, 2-trichloroethane
1,1,2,2-tetrachloroethane
chloroethane
bis (chloromethyl) ether
bis (2-chloroethyl) ether
2-chloroethylvinyl ether
2-chloronaphthalene
2,4, 6-trichlorophenol
parachlorometa cresol
chloroform
2-chlorophenol
1 , 2-dichlorofaenzene
1 , 3-dichlorobenzene
1 , 4-di chlorobenzene
3,3' -dichlorobenzidine
1 , 1-dichloroethylene
1,2-trans-dichloroethylene
i COl
iTLkCTOR
CATEGORY
/JB/1 NO.1
31A
32V
33V
34A
35B
36B
37B
38V
39B
40B
41B
42B
43B
44V
45V
46V
47V
48V
49V
50V
51V
52B
53B
54 B
55B
56B
57A
53A
59A
60A
COMPOUND jiz/1
2 ,4-dichloroph«»nol
1 . 2-dichloroprppane
1.2-diehloroprapvlene
2,4-dimethvlphennl
2, 4-dinitro toluene
2 , 6-dinitrotoluene
1 , 2-diphenvlhydrazine
ethvl benzene
f luorathene
4-chlorophenvl phenyl ether
4-bromophenyl phenyl ether
bis (2-chloroisoproovl) ether
bis (2-chloroethoxy) methane
methvlene chloride
methyl chloride
methyl bromide
bromoform
dichlorobromome thane
trichlorofluoromethane
dichlorod if luororae thane
chlorodibromome thane
hexachlorobutadiene
hexachlorocvclopentadiene
isophorone
naphthalene
nitrobenzene
2-nitrophenol
4-nitrophenol
2,4-dinitrophenol
4 ,6-dinitro-o-cresol
25
-------
SAMPLE I.D.
\
NO/ COMPOUND
61B Jll.Ili.t-£4.?.°ji. foe thy la mine
62 B N-nlirusod tplninyl.'imlnr
N-nitrosodi-n-p™
pentachlorophonni
henzo(a)anthracene
ben2o(a)j)yrene
3,^-benzoEJ uorathene
75B benzo(k)fJuoranthane
dlbenzo(a,h)anthracene
tetrachloroethvlene
crichloroethylene
2P-i COMPOUND
J8V vinyl chloride
aIdrin
*Not detected. =
tetrachlorodibenzo
pollueme...
26
-------
contractors and the regions comply with this format. The second, is a
description, provided by Athens, as to an acceptable format for the
preserved and archived GC/MS data tapes. This system is applicable to
a number of data units and is the one presently recommended by the
Agency.
Screening of Blanks
The analytical protocol provides the analyst with the option of
screening the blank samples on just GC prior to analysis. If he sees
nothing of significance for any of the compounds of concern he may
delete the requirement for looking at the total GC/MS run.
Analysis of Residual Chlorine
Concern was expressed at the meeting over the ability to measure free
residual chlorine in all waters as specified by the Agency. A
reference was sited which discusses the analytical problems revolving
around the application of residual chlorine analysis. At present the
Agency is looking into and attempting to evaluate the issue raised in
this particular area. As resolution is reached, the information will
be made available through the EPA Cincinnati office, the Environmental
Monitoring and Support Lab.
Metals Analysis
Introduction
The opening presentation on metals analysis was made by Dr. Fairless
of the EPA Region V S & A laboratory. Dr. Fairless presented a
program describing the various data that has been generated during
Phase I of the screening portion for a number of industrial
categories. The presentation and a synopsis of his slides are
presented in the next page.
Resolution of Phase II metals analysis is based on the suggestions
provided by the attendees and subsequent conversations.
A memo has been prepared and distributed by EPA headquarters, which
sets forward the procedures to be followed in the upcoming Phase of
the metals screening program. A copy of this memo which includes the
various industrial coding in contractor's codes is provided in the
reference section of this document.
27
-------
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«
-------
Acknowledgements
H. Montgomery
T. Meszaros
T. Parks
A. Jirka
G. Kunselman
E, King
M. Carter
J. Kirkpatrick
C. Elly
E. Huff
D. May
29
-------
General Observations
1. Samples are very variable in terms of relative concentrations
both within and between different industries.
2. Frequently a sample has at least one parameter with a very high
concentration relative to surface water and NPDES effluent dis-
charge samples.
3. As a result of the above facts, the samples are difficult to
analyze and the results show more scatter than is normal for
other sample types.
30
-------
Phase I - Quality Assurance
I General Operating Procedures
Samples were received unpreserved in different kinds of bottles.
Upon arrival at the laboratory, sufficient acid was added to each
sample to louder the pH to two. The sample was then allowed to
stand for several days.
Aliquotes were taken for analyses by flameless AA and ICAP.
Flameless AA analysis were made using standard addition techniques
on each sample. The ICAP method uses a standard EPA digestion.
Potassium dichromate is added to the remaining sample and an
aliquote taken for the mercury determination.
3]
-------
Quality Assurance
The CRL uses a semi-formal quality assurance program in which
selected performance audits are conducted to provide an estimate
of data quality. The following audits are run to monitor the
ICAP method:
Audit
1. Reagent Blank
2. Laboratory Control Standard
3. Sample Spikes
4. Reference Standards
5. Duplicate Samples
6. Duplicate Analyses
Frequency C%)
6
4
4
Monthly
32
-------
Ca*
Mg*
Na*
Ag
Al
B
Ba
Be
Cd
Co
Cr
Cu
Fe
Mn
Mo
Ni
Pb
Sn
Ti
Mean
20.6
4.8
16.9
153
922
452
952
81.5
427
442
300
492
2445
422
1102
513
5455
547
554
Laboratory Control Standards
for 77 different runs over 8 months
Sept. 76 - April 77
Std. dev.
1
0.4
1
23
68
37
43
2.7
12
16
14
21
119
13
42
16
306
23
19
Rel.Std.dev.(lZ)
5
8
6
15
7
8
5
3
3
4
5
4
5
3
4
3
6
4
3
33
-------
Laboratory Control Standards (cont'd)
Mean Std. dev. Rel.Std.dev.(1%)
V 533 53 10
Zn 2695 121 4
mg/1
34
-------
ok.
tic.
.ement #
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
Element
IS
TC
PM
B2
CA
CA2
MG
NA
AG
AL
AL2
B
BA
BE
CD
CO
CR
CU
FE
MN
MO
NI
PB
SN
TI
Detection
Limit (jpft
\,
0
0
0
0
7
5
1
15
1
50
50
50
5
1
2
5
5
6
170
5
5
5
20
5
15
Priority
0
0
0
0
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
AQC Spike
Level
0
0
0
0
10
10
10
10
0
1000
1000
400
1000
100
400
400
400
400
1250
400
400
400
400
400
400
35
-------
cont'd
ment #
26
27
28
29
30
Element
V
Y
ZN
XX
V2
Detection
Limit
12
16
60
20
1000
Priority
1
1
1
0
0
AQC Spike
Level
400
400
400
0
400
36
-------
7
Separate samples were collected for mercury and the other metals at
the beginning of the program. It is my understanding that the mercury
sample was a grab and the ICAP sample was a composite in at least some
of the cases. We tried to evaluate the data obtained from these
"duplicate samples" as shown below.
37
-------
Log
Number
17051
52
53
54
58
17094
5
6
7
8
17130
1
4
6
17002
08^
01
00
04i/
17122
3
Metal Mg
Magnesium
Result
468
180
11.9
214
233
17
14
14
13
69
28
27
18.7
KO.l
17
6-
9 "',,-•
6
19" X
129
150
Resul t
453
237
10.6
247
237
17
14
13
13
69.
19.5
19-
18.4
KO.l
19
19
5.8
~~V- 6
'6
128
140
Log
Number
17056
55
57
59'
60
17099
100
101
102
103
17133
32
35
37
17007
09--
03
06
05 -"
17126
7
38
-------
Metal Mg
Magnesium (cont'd)
Log
Number
4
5
17110
1
2
3
4
5
17043
44
47
49
17104
5
6
Result
340
10
284
340
238
350
191
4
14
52
22
13
9
19
19
Result
K300
11
192
193
166
208
195
5
14
52
, 22
13
14
20
19
Log
Number
8
9
17116
7
8
9
20
21
17045
46
48
50
17107
8
9
39
-------
Element
Al
Sb
As
B
Ba
Be
Cd
Ca
Co
Cr
Cu
Fe
Pb
Mg
Mn
Hg
Mo
Ni
Se
Ma
Sn
Ti
V
Zn
N
10
17
35
34
22
5
4
35
9
20
18
35
9
34
19
23
76
6
33
36
2
8
10
32
Ave
RPD
27
33
30
20
17
15
14
15
20
21
33*
32
13
10
23
24
19
- 21
33
15
27
36
15
21
Std. dev.
RPD
27
20
23
25
18
16
19
22
22
20
29*
24
11
14
20
21
23
11
24
19
10
24
13
25
40
-------
It is obvious from the data shown above that there is considerable scatter
in the "duplicate" sample results. This scatter may result from -
Analytical method
Samples are not true duplicates
Normal Sampling & Analytical Precision
a
From a comparison with our method performance audit results shown previously
it would appear that scatter due to the analytical method is a minor part
of the total data scatter.
-------
Phase II
1. We believe increased efforts should be made to improve and define
data quality for this project. As a minimum we recommend the following:
a. All contractors should use the same sampling procedure and sample
bottle type.
b. All samples should be field preserved and reagent blanks taken for
each survey.
c. All samples should be identified with a single 6 digit number.
d. Ten percent of all sites should be sampled in duplicate and one
member of each duplicate pair should be analzyed in duplicate.
42
-------
The steam electric metals data is available on request. These
particular analysis were performed on split samples taken during the
same relative time period and analyzed by both Carborundum, NUS and
EPA. In addition, EPA, early on in the program made an error in its
sample labeling procedure and forwarded two samples to the regional
lab, one a grab and the other a composite. The data therefore for EPA
will compare both a composite and a grab sample taken from the same
source. This data is provided for your information and scrutiny.
The comparative data of EPA and Mobil is presented in the comment
section. These samples were taken during the screening phase of the
review of the petroleum refining industry.
Also available on request is a comparison of the metals analysis
performed on samples collected from various coal mine discharges.
These analysis were performed by EPA's Region V laboratory; Bituminous
Coal Research in Monroeville, Pennsylvania, which used atomic
absorption spectroscopy for its method and Versar, Incorporated of
Springfield, Virginia, which also used atomic absorption. In
addition, Peabody Coal Company analyzed similar sets of samples taken
during the same time frame and this data may be provided. Moreover, a
number of samples were supplied to Gulf South Research and they were
analyzed both for organics by GC/MS and for metals by Spark Source
Emissions Spectroscopy. All of these samples were taken during the
same sampling period or are results of direct splits in the field.
Therefore, tabulation of the coal data lends itself to the closer
scrutiny of the metals analysis in general.
Issue: Digestion procedure for total metals.
A great deal of controversy has arisen over the application of "hard
digestion" that is digestion in nitric acid to almost dryness and
subsequent dilution in hydrochloric acid. A number of procedures
described what would be called a soft digestion or a sulfuric leach
discussions proceeded around this question and is included.
Resolution:
During Phase II of the screening and verification procedures all metal
samples will undergo hard digestion by nitric and hydrochloric acid.
They will not be taken to dryness as had been previously described due
to the number of comments received on the possible loss by
volatiliation or splattering of the sample in preparation.
Issue: Sampling Containers and Storage Bottles
43
-------
Comments were received in regard to the increase in mercury level in a
number of plastic containers.
Resolution:
Region V s S & A laboratory has carried out a number of studies
relating to storage containers and as a result has recommended the
following: Cap-while 43 400 MM, H-43 polypropylene smooth edge
linerless cap-W. Braum Co., 300 North Canal St., Chicago, IL 60606,
312/FI6-6500, container -125 cc or 360 cc wide mouth, oblong
polyethelyene (Monsanto) for use with 38-400 MM screw cap or 960 cc
wide mouth for use with 43-400 MM Cincinnati Container Co. , 2833
Spring Grove Ave., Cincinnati, OH 45225, 513/542-1515.
Issue: Preservation of metals samples
Resolution:
Based on the comments received, and in particular, a comment submitted
by Calspan, which outlined their attempt at obtaining a variance from
the DOT regulations, it has been decided that for Phase II as in Phase
I metal samples will not be acidified (preserved) prior to shipment.
Included is the letter submitted by Calspan to the Department of
Transportation requesting a variance and stating their case.
Furthermore there is included the subsequent denial by DOT to this
request (see the comment section) .
Asbestos Measurement
EPA (Athens) Presentation
EPA discussed their method for analyzing asbestos (chrysotile fibers).
This interim method is available in the reference section. This
method is scheduled for updating in August 1978, EPA has obtained
precise results with this method. A complete study of sample
preparation techniques by the Ontario Research Foundation will be
available in 3 months. EPA (Cincinnati) is preparing standards for
chrysotile fibers.
Calspan Presentation
Calspan has been performing chrysotile fiber counts and total fiber
counts on samples from the ore mining and coal mining industries using
the EPA analytical method. Calspan outlined a number of problems they
have encountered in these analyses including:
44
-------
Background fiber levels tend to vary in the diluent. They
recommend using a non-aqueous diluent, i.e. methanol.
Fibers have been found to protrude through holes in the nucleopore
filter.
A double carbon coating may be needed on the nucleopore filter.
With some fibers only partial SAED patterns or no patterns can be
seen.
The amount of solids in some samples hindered analyses.
Q(Versar): What are background levels of asbestos in U.S.
waterways?
A(Calspan) : Little data exists; the available literature indicates
these levels are between 105-107. Calspan recommends
verification sampling for waste streams containing more
than 1 x 10s chrysotile fibers.
A(McCrone) : Reported 10a-109 background levels in California.
Q(EPA Athens) Questioned the use of the exponential form for reporting
data they had selected the unit "million fibers per
liter (mfl)".
A (Calspan) : Responded that they agree that this unit should be
standardized.
Comments:
McCrone Compared the use of light microscopy with electron microscopy
for asbestos analysis. Asbestos fibers larger than 0.25 in diameter
can be seen, but smaller fibers (which are usually more numerous) are
missed. Many more fibers can be seen with transmission electron
microscopy. Using a TEM, fibers can be identified by electron
diffraction, chemical information, as well as by morphology. McCrone
commented that they had had problems with achieving a representative
distribution of fibers on the particle/grid square (see the reference
section).
EPA (Duluth) Presented slides and discussed the analytical method for
asbestos. Commented that double carbon coating of nucleopore filter
may distort the fiber and add to problems with the electron
45
-------
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TABLE 6. CHRYSOT1LE FIBER COUNTS EXPRESSED IN TERMS OF MASS FOR
ORE MIMING AND DRESSING FACILITIES
FACILITY
ALCOA
ASARCO-GALENA
ASARCO-GALENA
KENNECOTT-SLC
KENIMECOTT-SLC
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PLACE R-AIWEX
MclNTYRE DEVELOPMENT
PINE CREEK-UCC
MOLYCORP-QUESTA
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Al
Ag
Aq
Cu (02B)
Cu (048)
Cu (06B!
Cu (088)
Cu
Cu
Cu
Pb/Zn
Pb/Zn
Pb/Zn
Fe
Fe
U
U
U
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Hq
Ti
W
Mo
WASTEWATER SOURCE
TREATED MINE WATER
TREATED MINE WATER
TAILING POND EFFLUENT
TAILING POND EFFLUENT
TREATMENT PLANT EFFLUENT
TAILING POND EFFLUENT
TREATMENT PLANT EFFLUENT
TREATMENT SYSTEM EFFLUENT
TAILING POND EFFLUENT
TREATED MINE WATER
TREATMENT SYSTEM EFFLUENT
TAILING POND EFFLUENT
TAILING POND EFFLUENT
MINE WATER SETTLING POND
TAILING POND EFFLUENT
EFFLUENT FROM MILL SETTLING POND
TREATED MINE WATER
TREATED MINE WATER
TREATED MINE WATER
r TAILING POND RECYCLE
MILL WATER TO RECYCLE
TREATED MINE WATER
TAILING POND EFFLUENT
APPROXIMATE
CHRYSOTILE
MASS
(NANOGRAM/D*
70
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64
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"NANOGRAM/LITER = 10'9 GRAM/LITER
49
-------
diffraction pattern. In addition, in many industrial effluents,
organics may cloud or distort the electron diffraction pattern.
Commented that small fibers are difficult to identify by diffraction
pattern. Therefore they are often classified as ambiguous.
Discussing interlaboratory comparison of results, EPA commented that
there has been considerable improvement in the last few years. Labs
agree often on trends, but not on the magnitude on concentration.
Commented that long storage times hinder asbestos analysis. Fibers
tend to settle out and clump together and they are difficult to
redistribute.
Questions:
Q(Carborundum) : What is the minimum number of fibers which should
be counted?
A(McCrone) : Depends upon the background levels of asbestos. Total
suspended solids level also should be considered.
Q(RETA): What type of containers should be used for asbestos
samples? How long can samples be stored?
A(EPA Duluth): Recommend 1 liter samples be collected in poly-
ethylene containers. HgCl is recommended as an
antibacterial agent. Samples should be immersed
in an ultrasonic bath to prevent clumping. Samples
should be filtered as soon as possible. They can
be stored a long time but the container should be
placed in an ultrasonic bath to disperse fibers.
EPA does not recommend the use of a dispersal
agent because it tends to break up fibers, resulting
in a larger count.
Conclusion:
It is this Agency's position to recommend that the Transmission
Election Microscopy method, as described by Dr. Charles Anderson, EPA
(Athens) be utilized. In addition, a copy of the phamphlet Selected
Silicate Minerals and their Asbestiform Varieties is provided with
this package. This phamphlet is intended to serve as a basic source
of information. We are thankful to William J. Campbell of the Bureau
of Mines for supplying this publication.
50
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Biological Monitoring
In an attempt to present some alternative or perhaps a surragate
technology related to monitoring for organic or (possibly toxic)
contaminants, a discussion was held on the use of biological
organisms. A description was provided of the concept of the toxic
lethal units as related to biological monitoring.
Mr. Rawlings of Monsanto Corporation presented a description of the
present environmental assessment being carried out by Monsanto. This
study relates to the reduction of toxic effluents from the textile
industry. The Monsanto Program outlined the various stages of
activities that are ongoing in the various biological and chemical
test being employed to determine both the toxicity as well as the
treatability of a number of effluents coming from various textile
facilities. A brief description is provided along with the summation
of the slides that were shown.
-------
BIOASSAY TESTING OF INDUSTRIAL WASTEWATER
Presented to
EPA SEMINAR ON ANALYTICAL METHODS
November 9-10, 1977
Denver, Colorado
by
Gary D. Rawlings
Senior Research Engineer
MONSANTO PESEARCH CORPORATION
1515 Nicholas Road
Dayton, Ohio 45407
52
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Nutrients
Ammonia
Nitrite
Nitrate
Total Kjehldeh! nitrogen
Orthophosphate
Total phosphorus
Total organic carbon
Criteria Pollutants
BOD5
COD
Color (APHA)
Sulfide
Phenol
Total suspended solids
PH
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EPA TASK OFFICER
MRC PROJECT LEADER
SAM RLE COLLECTION
MRC
TESTING
-
MUTAGENICITY
SRI
CYTOTOXICITY
NORTHROP
FATHEAD MINNOW
AND DAPHNIA
EPA
FRESHWATER ALGAE
EPA
SHEEPSHEAD MINNOW
AND GRASS SHRIMP
BIONOMICS
MARINE ALGAE
EPA
ACUTE TOXICITY-
14- DAY RAT TEST
LITTON 3IONETICS
SOIL MICROCOSM
EPA
EPA -TECHNICAL
ADVISOR
HERL - RTP
ERL - NEWTON
ERL- GULF BREEZE
HERL- CINC
EPA -CORVALLIS
58
-------
59
-------
SAMPLING REQUIREMENTS
Biotest
Ames test
Cytotoxicity
Fathead minnow
and daphnia
Freshwater algae
Sheepshead minnow
and grass shrimp
Marine algae
14 -day rat test
Soil microcosm
Volume, liters
0.25
0.25
60.0
9.6
60.0
9.6
1.0
0.1
TOTAL 146.8
60
-------
MICROBIOLOGICAL MUTAGENICITY
Purpose: To determine if a chemical mutagen is present
in the wastewater.
Test species.- Salmonella typhimurium (Ames test)
Saccharomyces cerevisiae D3 (yeast)
Escherichiacoli WP2
Bacillus subtilis
Method: Subject each bacteria strain to the maximum
dosage allowed by the test, if a positive response
(mutagen present) occurs then proceed with
a dose response test.
-------
CYTOTOXICITY TEST
Purpose: To measure quantitatively cellular metabolic
impairment and death resulting from
exposure in vitro to toxicants.
Test species.- Rabbit alveolar macrophage
Method: Dose response test
62
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Teflon Lined
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Leachate Collection
Tube
66
-------
UNITS OF MEASURE FOR ACUTE TOXICITY
EC50; Effective concentration at which 50% of the test organisms
reach the desired effect. The "effect", for example, can be
growth inhibation or stimulation.
LC5Q.- Lethal concentration at which 50% of the test organisms
die over a specified time period (usually 48 or 96 hours).
LD5Q: Lethal dose at which 50% of the test organisms die
over a specified time period (usually 14 -days).
67
-------
Concentration: refers to the amount of sample (or toxicant) per unit
volume of test solution. Used,for exapmle, with fish
and shrimp tests.
Dose: refers to the measured amount of sample (or toxicant)
that was fed to the test organism. Used for the
14-day rat test.
es
-------
EXAMPLE
EC 50 = 10.2% This means that a 50% increase (or decrease) in the algal
mass occurred when exposed to a solution containing
only 10.2% of the secondary wastewater.
= 80.7% This means that 50% of the fathead minnows died when
placed in a tank containing 80. 7 % secondary wastewater.
Note: These toxicity values are determined graphically
from a series of dose reponse tests.
69
-------
code
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N.D. N.D. N.A.T.C
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41.0
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f
ne Ecoloa/ Tests
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12.8 90 1.3 MO
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MOO 59 1.3 MO
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71
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MRC/EPAWastewater
Toxicity Study:
Phase I: Screening
Collect Secondary Effluent Samples
from Each of the 24 Plants
Perform Analyses
Bioassays
Priority
Pollutants
Level 1 Chemical
Analysis
Evaluate Analytical Procedures
and Make Recommendations
for Improvement
Prioritize Plants
Based on Bioassay Toxicity Data
Select the Plants which have
Secondary Effluents Sufficiently
Toxic to Evaluate the Effect
of BAT Systems
72
-------
PRIQRITZATIQN OF TEXTILE PLANTS
_BY
TOXICITY OF SECONDARY EFFLUENT
Toxicity Ranking Plant
Most Toxic Group 1: A, B
Group 2: C, D, E
Group 3: F, G, H
Group 4.- I, J
Least Toxic Group 5.- K, L, M, N, 0, P, Q, R, S
73
-------
MRC/EPA Wastewater Toxicity StudyN
Phase II: j
Toxicity Removal by BAT Systems J
\
Select Bioassay Tests
Collect Sample for the Pilot Plant
During the 2 Weeks BAT Evaluation Period
Analyze Samples
Bioassays
Priority Pollutants
NO
YES
Report Results for BAT Evaluation
74
-------
BIOASSAYS USED FOR PHASE II
• Fathead Minnow
• Daphnia
• Freshwater Algae
75
-------
SIX TERTIARY TREATMENT UNIT OPERATIONS
1. Reactor/Clarifier (using combinations of alum, lime,
ferric chloride, and anionic and
cationic polyelectrolytes)
2. Multimedia Filter
3. Granular Activated Carbon Columns
4. Powdered Activated Carbon (laboratory test)
5. Dissolved Air Floatation
6. Ozonation
76
-------
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77
-------
i/l
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78
-------
INTERPRETATION OF BIOASSAY TEST RESULTS
Bioassay Results
Inlet Outlet
Toxic Substance Interpretation
Control Technology Is Not Effective
Control Technology Is Effective
Control Technology Is Deterimental
Control Technology Is Not Deterimental
79
-------
Index of Comments
1. Bruce Long and Carol Hammer, Ryckman, Edgerley, Tomlinson and
Associates, November 8, 1977, re: 0) Sampling and Shipment
(2) Metals Digestion, (3) VGA storage.
2. John Way, E.I. DuPont De Memours & Company, November 15, 1977,
re: Observations on the use of the E.P.A. sampling and analysis
procedures for priority pollutants - VOA - Acid and Base/Neutrals.
3. C.W. Phillips, Mobil Oil Corporation, November 2, 1977, re: com-
paritives data on analytical results for samples taken concur-
rently.
4. P.P. Hochgesang, Mobil Research and Development Corporation,
November 17, 1977, re: Acid Extractables - Phenolic compounds and
Base/Neutral - polynuclear aromatlcs.
5. M.J. O'Neal, Shell Development Company, December 15, 1977, re: VOA
apparatus, compositing and storage.
6. Dr. Robert Kleopfer,. U.S. Environmental Protection Agency, Region
VII, November 2, 1977. Comments concerning the experiences of the
E.P.A. surveillance and Analysis Division, when using the protocol,
7. Memo, dated January 18, 1978, Subject: Action Concerning Region
VII comments.
8. Letter: 6C-MS internal standards, Radian Corporation, Dr. Larry
Keith, November 21, 1977, corrected order of elution of the
unchlorinated base/neutral compounds.
9. P. Micheal Terlecxy, Calspan Corporation, December 19, 1977,
subject: Shipment of nitric acid and DOT regulations.
10. A.W. Garrison, E.P.A., Athens, Georgia, December 22, 1977,
Stability of two of the Consent Decree Pollutants.
80
-------
K YUM IIMII, CUOV5I U5T , l wi I tun
RDD RSSOCIRT6S
121(1 UckUnd feud
St. LaU«. MlMourt 13141
(314) 434-«MO
• dhrUlon ol Eniriradyn* Engineer*
November 8, 1977
Mr. William A. Telliard, Chief
Energy and Mining Branch
Effluent Guidelines Division
U. S. Environmental Protection Agency (WH-552)
Washington, DC 20460
Dear Mr. Telliard:
Ryckman/Edgerley/Tomlinson & Associates (RETA) appreciates
the opportunity to attend EGD's seminar on sampling and
analytical methods used for studies of priority pollutants
in industrial wastewaters. We feel this seminar is timely
and necessary.
RETA has been actively involved in the sampling and analytical
portions of BAT Review studies for both screening and verifi-
cation program in the Timber Processing, Petroleum Refining
and, currently, Organic Chemicals and Plastic and Synthetic
Materials Manufacturing point source categories. In the
course of our involvement in these programs (since January,
1977) we have learned many things which we would like to
pass on to your attention. Additionally, we have encountered
certain aspects of the most recent (April, 1977) protocol that
we feel require close attention with subsequent modification
and have developed several questions, the answers to which
are needed as soon as possible.
The points and questions we wish to pass on are the following:
I. Field Sampling, Sample Handling and Shipment
-i) Can Teflon sample tubing be reused?
_2) RETA is currently using Dow Corning selastic silicon
rubber medical grade 3/8-inch I.D. pump tubing
rather than tygon tubing.
3) The April, 1977 protocol requires a minimum
aliquot volume of 100 ml. Shouldn't this minimal
volume be 50 ml?
81
1
-------
, romunson
RSSoctRies
Mr. William A. Telliard
November 8, 1977
Page Two
4). 'jCiie composite metals sample is currently poured
fcff from the NVO composite sample. It is our
'opinion that a separate metals composite sample
should be collected by compositing grab samples in
a glass bottle with re-distilled nitric acid already
added.
5) ?ield experience has demonstrated that potassium-
iodide starch paper is not sensitive to residual }
chlorine levels as low as 2 mg/1. RETA is therefore 't
using the ortho Tolidine (OT) test in parallel •.
to the Kl-starch test. If the OT test is positive we
are adding 0.6 gm of ascorbic acid to the cyanide
sample and two drops of sodium thiosulfate to the
VOA samples.
6) Current protocol calls for analysis of both the
unpreserved and preserved VOA samples. RETA questions
the value of analyzing the unpreserved sample especially
when the cost of analysis is considered.
7' The April, 1977 screening protocol calls for preser-
vation for phenols "...by addition of phosphoric acid
or sulfuric acid to 4." RETA is adding 1.0 gram of
CuS04 followed by the addition of phosphoric acid to
lower the sample pH to 4.
8) Concerning sample blanks for VOA samples it is RETA's
interpretation of the April, 1977 protocol that two
VOA blanks are sent back per sampling site. These
VOA blanks are sent back with the NVO composite.
9) Concerning VOA samples, our experinece has found
vacuum bubbles formed in several VOA vials. We
believe this to be due to the cooling of the sample
with consequent volume reduction.
10) When several (more than one) sample sites require
compositing to comprise one composits sample, the
October, 1976 protocol permitted compositing of
VOA samples at 4°C No mention is made of this
procedure in the April, 1977 protocol.
82
-------
RYCKrriRn. eoeeruev, romunson
RFID RSSOCIRT6S
Mr. William A. Telliard
November 8, 1977
Page Three
IjL) RETA plans to performfGC/M^ analysis on the i
to treatment and ef f Iu5*3?""from treatment only^ GC_
analysis of the intake water will be performed" Iur"7
those priority pollutants found in the raw wastewater.
\
12) The April, 1977 protocol states, "When more than one
laboratory is involved. . .the sample should if at all
possible not be divided in the field but rather at
the contractors laboratory." RETA is pouring off
the metals fraction for those metals to be analyzed
in RETA's lab and preserving in the field using
re-distilled nitric acid. It is RETA's position that
not preserving as soon as possible increases the time
for metals adsorption on the NVO composite jug.
]p) PTTi*i rnr^iK ii i ji i n i i Ti ' n i 1 _^ri what constitutes
_ "analytical .method validation*7"^ called for in
memorandum of June 23, 1977 .
II . Analytical Considerations
1) The digestion procedures prescribed for metals. RETA
is concerned a) that the level of nitric acid prescribed
during digestion leads to low results for some of the
metals, and b) that the addition of the nickel nitrate
matrix must be done just prior to analysis for arsenic to
avoid loss of the metal, thereby precludiag__addition and
-storage for analysis of both selenium and arsenic.
2) Direct aqueous injection for acrylonitrile and acrolein
by GC/MS. Concern arises over the introduction of water
into the mass spectrometer.
3) Holding times for analysis. Concern arises over the
amount of time a^_vola_tile sample can be held prior_to
analysis and the^recommended storage alternatives" in "'
light of apparent problems arising from storage of
hermetically^eaTira trapgu^
•^ an~* —• "•
Thank you once again for the opportunity to attent this seminar
and for your consideration of the points raised in this memorandum,
Bruce W. Long, P.E. Carol A. Hammer, Ph.D,
83
-------
IC-MI'-E REV. 4-74
E5TAIUSHIO1802
E. I. DU PONT DE NEMOURS & COMPANY
INCORPORATED
WILMINGTON, DELAWARE 19898
INDUSTRIAL CHEMICALS DEPARTMENT
RESEARCH & DEVELOPMENT DIVISION
EXPERIMENTAL STATION
November 15, 1977
W. A. Telliard, Chief
Energy and Mining Branch, EGD
United States Environmental Protection Agency
ANALYSIS PROCEDURES FOR PRIORITY POLLUTANTS
In connection with our discussions at the Denver
Seminar, you may find the attached useful. Comment B-5, in
particular, refers to our experience with centrifugation as
a method of breaking emulsions.
Please let me know if you need further information.
JWW/td
Attach.
/ohn W. Way "'
Research Supervisor
84
BETTER THINGS FOR BETTER LIVING . . . THROUGH CHEMISTRY
-------
TAp
cc: JBColeman, ICO, W
RCOtt, ICD, W
GDBarbaras
Industrial Chemicals Department
Research & Development Division
Experimental Station
October 5, 1977
TO: J. W. WAY
FROM: R. T. ITEN
OBSERVATIONS ON TH
ANALYSIS PROCEDURES
INDUSTRIAL EFFLUEN1
USE OF THE EPA SAMPLING AND
F0R PRIORITY POLLUTANTS (PP's) IN
PS - EPA METHOD ISSUED 4/18/77
A. VOLATILE ORGANICS
1. Samples must be taken in specially cleaned glass bottles
with Teflon® lined caps and analyzed within 14 days (refriger-
ate) . No air bubbles are allowed in the bottle.
2. Pure water has to be prepared from distilled water passed
over absorbent chartoal and stored in special clean glass
bottles with Teflon lined tops.
3. Tekmar Concentrator trap supplied in some cases cannot be
used with the standard silica gel and Tenax GC - new columns
must be made using tenax GC only.
4. Tenax GC must be conditioned at least 16 hours at 350° C
(Tekmar conditions it at 200°C).
5. Samples standards and work-up must be done in a lab separ-
ate from the analysis and pure water prep area.
6. Blank Water (and tmre water from lab) sample must accom-
pany sample and be treated the same.
i
7. Tekmar should be kept at 250° in trap bake mode overnight
after running samples and also at least 15 minutes between
samples (also before initial use).
8. Desorption tubes (sample tubes) should be cleaned with
special water and baked 120°C between uses.
9. All gas supply lines should be cleaned and baked out before
installation and only ultrapure -Helium used.
85
-------
J. W. WAY - 2 - OCTOBER 5, 1977
10. Computer should have all PP's in search file.
11. All syringes should be cleaned only with the special high
purity water, or non-PP high volatility solvents.
B. ACID AND BASE/NEUTRAL SAMPLES (see also all of part A)
1. Require 2 to 4 liters for each type of analysis.
2. Require 2 liters blank water as reference.
3. No stopcock or other lubricants can be used anywhere in
the anlysis system. Only glass & Teflon® equipment may
be used. (As in part A, all supply gases must be pure
and delivered through clean gas lines.)
4. Sodium sulfate used for drying MeCl2 extracts should be
heated to 500°C for 2 hours and 1 Ib. for each sample
then washed with two 100 ml portions of MeCl2 which is
saved and analyzed as a composite. (Drying column is a
one liter cylindrical separatory funnel with glass wool
in bottom to hold 1 Ib. of Na2S04).
5. The MeCl2 extraction step results (probably in most cases)
in an emulsion which can only be broken by centrifugation
in closed centrifuge tubes (Teflon*5 seals). IEC HN-S centri-
fuge 1800 RPM 50 ml cups 30 min.
6. Combined extracts should be followed through ^3804 drying
column with two 100 ml portions of MeCl2 which is added
to total extract.
7. Note that all the precautions observed in A must also be
observed here. Standards, especially, must be prepared
under isolation and refrigerated as noted in procedures.
B. The use of a GC integrator is helpful in calculation of
the quantitative data.
86
-------
Mobil Oil Corporation ^
/ 'r/£/-)
' November 2, 1977
150 EAST 4?NO STREf
NEW YORK. NEW YORK 100U
Robert B. Sciiaffer, Director _
Effluent Guidelines Division (WK-552)
U.S. Environmental Protection Agency
401 M St., S.W.
Washington, D. C. 20460 cc: Loon H. Myers
Attn: Robert Dellinger Arnold S. Vernick
TOXIC SUBSTANCES SURVEY
MOBIL REFINERY DATA
(AUGUSTA, KS.)
Dear Mr. Dellinger:
We have completed our review of both the priority pollutant data
for "Refinery F" published in the September 30, 1977 Burns & Roe
preliminary draft report and the "Refinery 6" section of the
R. S. Kerr Environmental Research Laboratory screening survey
report on the petroleum refining industry.
The purposes of this letter are to 1) draw comparisons betv.een
the Burns S Roe results and the Mobil data obtained on separate
samples taken concurrently with the April 6-8, 1977 EPA sampling,
2) comment on certain observed deficiencies of the gas chromatog-
raphy/mass spectrometer (GCMS) analytical protocol utilized by
EPA, and 3) offer a rev; clarifying remarks for incorporation in
the final R. S. Kerr Laboratory report.
The attached Tables 1, 3 and 4 present comparative data for any
priority pollutant: detected at Augusta by either EPA or Mobil.
Table 2 provides an in-depth analysis (GCUV vs. GCMS) of the poly-
nuclear aromatics (PNA) group of priority pollutants found in the
base-neutral extractible semivolatiles.
Finally, our comparison and comments on analytical results for
eight NPDES pollutant parameters (Burns & Roe page IV-31, Table
IV-8) appears as the attached Appendix A.
Summary
e Table 1 presents comparative data for any volatile or semi-
volatile priority pollutant detected by either Mobil or EPA.
In general, the data presented in Table 1 for those compounds
that were detected by both Mobil and EPA agree to within a
factor of 3 which is quite reasonable considering the low
pollutant levels encountered.
87
-------
Mobil
-2-
Robert B. Schaffer
Director
November 2, 1977
Overall, Mobil analytical methodology for the semi volatile
pollutants enabled us to detect these pollutants at much
lower levels than the protocol used by EPA. The two major
factors contributing to this sensitivity advantage were (a)
PV(-r^p-*-inc a fafrror Trnl nmo nf T.rat-p-r (l^-?Cl liters VS. 2 liters),
(b) _evaporatinq more of the solvent to provide a more concen-
trated extract./ Thus, whereas the EPA procedure detected only
a total of five semivolatile pollutants in the Augusta water
samples, twenty semivolatile pollutants were observed by the
Mobil procedures. uith the exception of two phenolic pollutants,
all the priority pollutants observed by Mobil that were not
detected by £PA,_QCc.ur at concentrations less than 1Q ua^l.
Our major concern with the data reported by the EPA is the
apparent failure to recognize certain deficiencies of the GCMS
analytical protocol with regard to detection of several PNA
priority pollutants. For example, the EPA data state, without
any reservations, t-H^*-_ ^hrygpno anH bpn^o (a ) pyrr>no . BaP , are
found in all three Augusta water samples. We maintain that the
GCMS technique cannot unambiguously distinguish _phenanthren
f mm nT-^hrnccne , ch rvcpnp ">T-n-n h^n -70 i * } an f-h -r^ceno ( BaA) t or_"
a)ovrcno (BaP) from erlene
_
. The EPA data for the base
should definitely be amended to
pery-
_
neutral priority pollutants
reflect the facts that chrysene and BaA as well as Ba? and
lene arc indistinguishable. Mobil recognized the inadequacy
of the GCMS protocol for the PNA class of ^riority pollutants
and simultaneously carried out GCUV measurements on these comr
pounds . Table 2
EPA GCMS data._ ________ _ _ __
and the effluent from the "oxidation pond (EOF) sample", tffe
results obtained for benzo (a) pyrene by the more definitive GCUV
technique are lower by close to a factor of 10 than the GCMS
values .
compares "the GCUV results
'or bththe. coolinq tower
~th
with both
efjfl ue n t
(EOPT
Mobil and
CTE) sample
Specific Comments
Comparisons between Mobil and EPA (Burns & Roe) results are treated
below:
Methylene Chloride
EPA detected methylene
ug/1 in the IPBS, CTE,
EPA blanks from these_
chloride at levels of < 10, 70, and
-------
Mobil
-3-
Robert B. Schaffer November 2, 1977
Director
chloride at the 50, .— 10, and 50 ug/1 level. The presence of
methylene 'chloride in the EPA blanks at these levels suggests
contamination of the samples in either the sampling or the
analysis.
Mobil detected methylene chloride only in the IPBS sample at the
5 ug/1 level.
Carbon Tetrachloride
Carbon Tetrachloride was detected by EPA at a concentration of
greater than 50 ug/1 in the IPBS sample. No carbon tetrachloride
was detected by EPA in either the CTE or the EOP samples. The
EPA blanks were likewise free of carbon tetrachloride. Mobil
did not detect carbon tetrachloride in any of the Augusta samples
including the duplicate EPA samples supplied to us by the EPA
sampling team. It should be emphasized that the halogen selective
chromatoaraohic detector that wa^ r-irp i nyng tor tne vpTat-.il.pj
orcranirs anf\
uVSGS COU.
rl
ce
tect
carbon
tet:
*ach
.ori
de.
at t-hp
uq/
likely a result of contamination within their analytical laboratory,
1,1,1-TrichloroGthane
Agreement between Mobil and EPA for this priority•pollutant is
excellent. The compound was observed only in the IPBS sample -
Mobil obtained 55 ug/1, EPA reported greater than 50 ug/1.
Benzene,Toluene and Ethylbonzene
The agreement for these three volatile organics between Mobil and
EPA is juit^ rp^sonahlp. Neither Mobil nor EPA detected any of
these aromatic hydrocaroonsin the CTE or EOP samples. Mobil
reported levels of 6, 14 and •<. 0.5 ug/1 for benzene, toluene, and
ethylbenzene respectively in the IPBS sample, whereas EPA did not
detect any of these hydrocarbons. The reason for the discrepancy
on this site most likely is due to the fact that EPA utilized a
GCMS technique for identification and quantitation as compared to
the less specific flame ionization detector (FID) employed bv
~ln this regard, tne Mobil values should be considered
as upper limits.
89
-------
Mobil
-4-
Robert B. Schaffer November 2, 1977
Director
)
Acid Extractable Semi Volatiles
EPA did not detect any priority pollutants in the acid extractable
semivolatile category whereas Mobil observed three; namely, phenol,
2,4-dimethylphenoI, and p-chloro-m-cresol. Phenol was observed in
the Mobil EOF sample at a concentration of 59 ug/1. The EPA
i.-i nn l imits £nr the acid cxtractables. vary from 10 t-n JflO ua/1.
_ nrT oj-j the nature of the ofhqr fnnrH nnal.groups j For phenol
and 2,4-dimethyTpnenoi me detection limit is reported to be about
10 ug/1, and so EPA's inability to detect these components In the
IPBS and the CTE samples is understandable. The fact that they
did not see phenol in the EOF sample may be due to the manner in
which EPA prepared their composite sample prior to extraction. Un-
T1ivp fhr> Mnhil procedure f wherein all the water taken at a _
particular site over the three cay sampling period, was extracted.
the EPA protocol called tor withdrawing allquots from eacn ofthe
three 24-hour composite samples and then extracting this blended
composite. In this procedure trace components could be easily
lost to the glass walls of the original sample container, and we
would expect that adsorption of trace organics would be most severe
for highly functional compounds such as the phenols.
PNA' s
In addition to our major concern noted in the summary regarding the
ambiguities of the EPA GCMS protocol for accurate identification
of certain FNA's, there are other cases where Mobil & EPA data
differ by more than a factor of 3 that deserve comment. First, the
•Icbil and ETA GCMS data for thp__ iponpnn-^r: f.luoranthane in the IPBS
sample differ by close to a factor of 10. Mobil determined 3.4 ug/1
v/hereas EPA observed 29 ug/1. However, the Mobil GCMS value of
3.4 ug/1 is in excellent agreement with the Mobil GCLJV value of
4.6 ug/1. In a similar fashion the EPA level of 10 ug/1 for pyrene
in the CTE sample is higher than the Mobil value of 2.9 ug/1. Here
again the Mobil GCUV value of 2.6 ug/1 is in excellent agreement
with the Mobil GCMS result. These cases given added credibility to
the accuracy of the Mobil data.
Pesticides
The EPA subcontractor, Rykman, Edgerly, Tomlinson & Associates (RETA),
tentatively reported that
-------
Mobil'
-5-
Robert B. Schaffer November 2, 1977
Director
The following comments relate to the R. S. Kerr Laboratory draft
report:
1. Refinery =6, page 2 - penultimate paragraph, last sentence:
Replace "Sludge from .... soil farming" with "Oil recovered
from the oil skimming pond is treated to remove recoverable
oil. Sludge from the treating process is used for soil
farming".
2. Figure 2, title should read "REFINERY WASTEWATER FACTLITIES"
Please feel free to contact me if you have any questions.
SM Jack son /Yah
Attachments
. 1 1-'- -/yi.«. /
C. W. Phillips
91
-------
A"i'ist'i Refinerv TOXIC ?ubst.Tnc«s Survev
Comparison of Mobil and EPA
Data for
Priority
IPI3S
Compound Noire (s)
Volatile Organics
Benzene
Toluene
Ethylbenzene
Carbon Tetrachloride
Methylene Chloride
1,1, 1 -Tr ich lor oe thane
Acid Extractables
Phenol
2, 4 -dimethyl phenol
p-chloro-m-cresol
Base Neutral Extractablcs
bis (2-ethylhexyl) phthalate
(a)
PNA's ' '
Naphthalene , .
( c ]
Acenaphthylene
Acenaphthene
Fluorone
Phenanthrene + Anthracene
Fluoranthene
Pyrcne
Chrysene -*• Benzo (a ) Anthracene
Benzo (b) Fluoranthone + Eenzo (k) Fluoranthene
Benzo (a) Pyrene + P°rylcne
Indeno (1,2, 3-cd ) Pyrer.e
Dibenzo(a,h) Anthracene >c '
Benzo (g, h, i) Pery lene
Pesticides
BKC
Mohi 1
ug/1
6
14
< 0.5
ND
5
55
4
5
ND
5
3.6
ND
5
9.6
146
3.4
65
76
6.4
54 (b)
0.3
ND
5
ND
FPA(d)
ug/l
ND
ND
ND
> 50
< 10
> 50
ND
ND
ND
ND
ND
ND
ND
ND
164
29
143
48
ND
33
:.T>
ND
ND
Pollutants
CTE
Mob i 1
ug/l
ND
ND
ND
ND
ND
NT)
6
11
ND
2.2
ND
ND
ND
ND
4.4
ND
2.9
16
1.8
16
1.0
KD
2.4
ND
EPA(d>
ug/1
ND
ND
ND
TO
70
ND
ND
ND
ND
ND
KD
ND
ND
ND
1.8
ND
10
6.5
ND
9.5
ND
;ro
ND
Mobil
ug/l
ND
ND
ND
ND
ND
ND
59
8
'0.2
10
0.1
ND
NT)
ND
0.4
0.2
0.5
1.4
< 0.2
2.2
ND
NO
ND
ND
EOP
FPA(d)
ug/l
ND
ND
ND
ND
< 10
ND
ND
ND
ND
ND
NT)
ND
ND
ND
:;D
ND
ND
0.8
ND
1.3
ND
ND
ND
trace^
(a) Mobil Data on PNA's is data determined by the GCM? technique.
(b) For this site only. BaP and perylene were individually determined by GCMS,
BaP = 16 ug/l, perylene = 38 ug/l•
(c) This component was detected by GCUV.
(d) Data obtained by Burns & Roe.
(e) Preliminary EPA data that is based on GCEC.
92
-------
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93
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TABLE 3
ANALYTICAL RESULTS FOR PRIORITY POLLUTANTS
CYANIDES, PIIENOLICS, & MECURY
PAGE IV-41, TABLE IV-13, BURNS & ROE DRAFT REPORT
Cyanides mg/1
Intake
CTB
B&R L0.03
Mob 0.00 average
0.52 to 0.83
0.02 average
Final Effluent
0.06 to 0.08
0.02 average
Mobil results indicate much lower concentrations than Burns & Roe
report. We believe this is significant and recommend that addi-
tional sampling and analysis should be carried out prior to state-
ment of the cyanide content of water a Refinery F.
Phenolics mg/1
Intake
CTB
B&R
Mob
0.21
0.190 average
0.042 to 0.056
0.037 average
Final Effluent
0.023 to 0.056
0.015 average
The interlaboratory agreement is acceptable.
Mercury mg/1
Intake
CTB
B&R
Mob
.0002 to .0009
0.000
.0004 to .0007
0.000
Final Effluent
.0003 to .0004
0.000
Mercury content is shown to be less than one part per billion by
all analyses of Refinery F water samples.
94
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TABLE 4
ANALYTICAL RESULTS FOR PRIORITY POLLUTANTS
METALS
PAGE IV-46, TABLE IV-14, BURNS & ROE DRAFT REPORT
B&R
Mob
Intake
L5 to L250
ND
Silver ug/1
CTB
L5 to L250
ND
Silver was not detected in any analysis.
B&R
Mob
Intake
L2 to L26
ND
Beryllium ug/1
CTB
L2 to L3
ND
Beryllium was not detected in any analysis.
B&R
Mob
Intake
LI to L200
ND
Cadmium ucr/1
CTB
LI to L20
ND
Cadmium was not detected in any analysis.
Intake
Chromium ug/1
CTB
B&R
Mob
60 to 72
59 average
44 to 79
38 average
Final Effluent
L5 to L25
ND
Final Effluent
L2 to L3
ND
Final Effluent
LI to L20
ND
Final Effluent
7 to 73
15 average
Interlaboratory agreement is acceptable but range of Burns & Roe
results for final effluent is rather high.
Intake
Copper ug/1
CTB
B&R
Mob
50 to 210
201 average
278 to 510
377 average
Final Effluent
84 to 199
98 average
Interlaboratory agreement is acceptable.
95
-------
TABLE 4 (cont'd)
Intake
Nickel ug/1
CTB
B&R
Mob
57 to 62
40 average
64 to 134
98 average
Final Effluent
58 to 74
58 average
Interlaboratory agreement is acceptable.
Intake
Lead ug/1
CTB
B&R
Mob
L15 to L600
3 average
L15 to LGO
5 average
Final Effluent
L15 to L60
1 average
Interlaboratory agreement is acceptable.
Intake
Zinc ug/1
CTB
B&R
Mob
120 to 133
217 average
229 to 452
300 average
Final Effluent
100 to 151
177 average
Mobil results nay be somewhat high which suggests rechecking
sampling and calibration.
B&R
Mob
Intake
27
15 averate
Arsenic ug/1
CTB
41
24 average
Final Effluen_t
31
16 average
Interlaboratory agreement probably is acceptable although Burns &
Roe results generally are higher than Mobil.
B&R
Mob
Intake
L25
6 average
Antimony .ug/1
CTB
L25
8 average
Final Effluent
L25
9 average
Interlaboratory agreement is acceptable.
96
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Mobil Research and Development Corporation
RESEARCH DEPARTMENT
PAULS80RO, NEW JERSEY 08066
C. H. UECHTHALER
MANAGER
PROCESS RESEARCH AND
TECHNICAL SERVICE
November 17, 1977
Mr. William A. Telliard
Energy & Mining Branch/EGD (WH552)
U.S. Environmental Protection Agency
Room 907, East Tower
401 "M" Street, S.W.
Washington, B.C. 20460
Dear Mr. Telliard:
EPA SEMINAR ON ANALYTICAL METHODS
DENVER, COLORADO
NOVEMBER 9-10, 1977
As you requested, I have prepared the enclosed amplification
of comments I made at the Denver seminar. Please feel free
to contact me if you have any questions.
Very truly yours,
mm
enclosure
cc: P. L. Gerard
C. W. Phillips
F. P. Hochgesang
97
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Supplementary Comments to Verbal Presentation by F. P. Hochgesang
at EPA Seminar on Analytical Methods, November 9 & 10, 1977
Agenda Item IIA2
Acid Extractables - Phenolic Compounds
2,4-Dimethylphenol is only one of a group of 9
possible C-2 substituted phenolic compounds which may be
found near the same GC/MS scan number. These phenolics are
poorly resolved on packed GC columns. A SCOT column (70
meters x 0.5 mm ID glass; SE 30, silanized and deactivated)
does resolve the unknown mixture to the extent that five peaks
sometimes appear. The tentative identification of these five
peaks in order of elution from the above SCOT column is:
1. 2,6-Dimethylphenol
2. 3.5-)- 2,3-Dimethylphenol and 3 + 4-ethylphenol
3. 2,4 + 3,4-Dimethylphenol
4. C-2 alkylphenols
5. C-2 alkylphenols
Base/Neutral Extractables - Polynuclear Aromatics
Benzo (a)pyrene usually is found in the same GC/MS
scan as perylene. GC/UV can distinguish each of the above PNA
isomers and our experience indicates that perylene usually is
present in higher concentration than benzo (a)'pyrene. Other
PNA's also appear as pairs when the EPA analytical protocol is
used. Therefore we suggest that the component names applied to
the EPA GC/MS protocol should be as follows:
Anthracene + Phenanthrene
Benz (a)anthracene + Chrysene
Benzo (a)pyrene + Perylene
Benzo (b)fluoranthene + Benzo (k)fluoranthene
Further, experience to date indicates that the
concentrations of fluoranthene and pyrene as determined by the
EPA subcontractor are higher than those values determined by
Mobil. The Mobil data for these components when determined by
both GC/UV and GCMS are in excellent agreement, but have been
found to be 4 to 8 times lower than the EPA GC/MS values.
We do not know the reason for these differences between the
Mobil and EPA values for fluoranthene and pyrene at this time.
We suggest that spiked samples be studied carefully in the
validation and verification phases.
98
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SHELL DEVELOPMENT COMPANY
A DIVISION OF SHELL OIL COMPANY
WESTHOLLOW RESEARCH CENTER
P. 0. Box 1380
Houston, TX 77001
December 15, 1977
Mr. William A. TeTHard
United States Environmental Protection Agency (WH552)
401 M St. S.W.
Washington, D. C. 20460
Dear Sir:
We are appreciative for the opportunity to have had
P. A. Wadsworth and G. H. Stanko participate in the EPA Effluent
Guidelines Division seminar on November 9 and 10 in Denver, Colorado.
At the seminar, some analytical work that had been done
on compositing samples in the VOA apparatus and on the effect of
storage time for VOA (vials) samples was disclosed. Also discussed,
were a number of problems and modifications that have been made to
the Tekmar LSC-1 unit. Enclosed, per your request are some of the
data relating to the VOA analytical procedure and a schematic of
some of the modifications made to the LSC-1 unit.
Attachment I and II indicate that one can composite VOA
samples in the apparatus. The compositing technique reduces the
number of individual analyses that are required to characterize a
wastewater sample for a 24-hr period. However, Attachment II shows
one of the disadvantages of the compositing technique. The individual
VOA analyses revealed short term differences that probably would not
have been detected by the compositing technique. Attachment II also
indicates some of the potential problems of taking single grab samples
in an attempt to define typical operations.
We have very limited data available on the storage of
petroleum/chemical wastewater samples in VOA vials. Normally, our
VOA samples are analyzed within three days from collection. Manpower
limitations and backlog did not allow for a more thorough investigation.
We collected a number of vials of a wastewater and ran VOA by GC/MS on
days 0, 7, and 20. Examination of the mass spectra showed significant
reductions in concentrations for many of the compounds for the sample
run on the 20th day of the study. No significant differences were
observed for the 7th day sample. The EPA Laboratory (EMSL) at
Cincinnati indicated VOA vials can be safely stored at 4°C up to 14
days. The limited data that we have appears to be consistent with
the Cincinnati recommended storage time.
99
-------
Attachment III is the schematic which shows the modifications
that have been made to the Teckmar LSC-1 unit. A number of additional
changes were made that are not shown on the schematic. The brass/0-ring
fittings on the sampler (glass) have been replaced with stainless steel
fittings with Teflon ferrules. This change was necessary to eliminate
a memory effect that was being observed. The samplers are cleaned off-
line and then vacuum baked. Intractably dirty samplers are discarded.
In order to eliminate still additional sources of memory effects, the
stainless steel tubing and the six-port valve are all heated with
electrical heating tapes. The temperature of the ooints indicated
are monitored with thermocouples using a Doric 402A-J/C Trendicator.
We believe that our future participation in seminars such
as was held in Denver to be of significant value to all concerned and
we wish to again express our appreciation for the opportunity to have
recently participated in such a seminar.
Very trul
GHS/PAW/thc M. J. O'Neal, Manager
Analytical - Chemical/Oil Department
cc: Judith G. Thatcher - API/DEA
Carl A. Gosline - MCA Staff
Ron 0. Kagel - MCA/Dow
100
-------
ATTACHMENT I
TEST OF COMPOSITING SAMPLE IN VGA APPARATUS
SAMPLE: REFINERY WASTEWATER
CONCENTRATION, PPM. MT.
SAMPLE _A_ _B_
1430 3/15/77 4.50 2.83
2230 3/15/77 5.50 3.20
0630 3/16/77 6.36 5.79
AVERAGE 5.45 3.94
SAMPLE COMPOSITED
IN VOA APPARATUS 5.83 3.02
101
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ATTACHMENT II
TEST OF COMPOSITING SAMPLE IN VOA APPARATUS
SAMPLE: PROJECT G
DAY 1 4/11
DAY 2 4/12
DAY 3 4/13
AVERAGE
SAMPLE COMPOSITED
IN VOA APPARATUS
COMPOUND CONCENTRATION, PPM. HT.
ABC
489. 0.081 0.063
370. 0.051 0.018
74. 0.011 0.010
310 0.048 0.030
354
-NOT CALCULATED-
103
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ATTACHMENT III
TEKMAR LSC-1 MODIFICATIONS
Regulated Helium Supply from
GC on GC/MS
Gas Chromatograph or
Gas Chromatography/
Mass Spectrameter
Return Line
from Tekmar
Desorb Gas and Sample Out
V
Flow
Controller
01724
104
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UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
SUBJECT:
FROM:
TO:
Comments on "Sampling and Analysis Procedure for DATE: - November 2, 1977
Screening of Industrial Effluents for Priority Pollutants"
Robert D. Kleopfer, Ph.D., Chief, Organic Chemistry Section, SVAN-LABO
Charles P. Hensley, Acting Chief, General Analyses Section, SVAN-LABOC/5^
William J. Keffer, P.E., Chief, Water Section, SVAN-TECH
Files
Based en three months experience with sampling and analysis for the
129 priority pollutants, the Region VII Surveillance and Analysis
Division has several comments to make concerning the guideline document
which was provided by EMSL. Overall we feel that the recommended
analytical protocol which was provided is a sound one. However, we
did find some deficiencies and errors. Also, we have included some
comments concerning the Radian consent decree standards which were
provided by the Environmental Research Laboratory in Athens.
Organ:cs by Purge and Trap
The recommended packing material as supplied by Supelco (Carbopack C/
0.2%, Carbowax 1500 with a Chromosorb W/3% Carbowax 1500 precolumn)
is not adequate for the analysis of chloromethane and dichlorodi-
fluronetnane which are the two most volatile compounds (boiling
points of -24°C and -30°C, respectively) on the priority pollutant
list. Discussions with Tom Bellar of EMSL indicate that the problem
relates to variability in the quality of the packing material from
batcn to batch. Indications are that a batch which performs adequately
has ^o: yet been located.
Assunnng an adequate analytical column were available, a modified
purger "netnod would be required for the analysis of dichlorodifluro-
methare. In effect, this doubles the amount of time required to do
all cf the purgable compounds (from about 45 minutes to 90 minutes).
We suggest that this compound be searched for only in selected samples.
Bis(crrlorcmethyl )ethsr cannot be analyzed by the purge and trap procedure.
As nc-=£ in Table I of the procedure, the compound has a very short
half-life in water and would not likely be found in a watar sampls
anyway, considering that samples are at least 24-hours old before
analysis begins.
In our laboratory, we have a difficulty with a methylene chloride
background because of the large amounts of this solvent which are
utilized in our extractions. In order to effectively deal with this
problem, we store all of our volatile samples in sealed dessicators
containing activated carbon. Nonetheless, we do routinely observe a
methylene chloride background in most of the samples which we run.
However, the background amounts to less than 20 PPB which is our
reported detection limit.
EPA Farm 1320-6 (R.v. 6-72)
105
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Although in most instances for the purge and trap technique we can
analyze successfully at concentrations much lower than 20 PPB, we are
using that value as our lower detection limit. When compounds are
detected at below that amount, it is considered a trace amount and
indicated as such on the data sheet. Some of-our recovery data for
the purgable compounds are attached.
The mass spectrum for 1,1,1-trichloroethane does not have a base
peak at mass 98 as indicated in Table II. The base peak is actually
at mass 97.
Base/Neutral Compounds
An incorrect mass is given in Table IV for diethylphthalate. The
corrscc nass for one of the characteristic El ion should be 177 rather
than 173. Mass 177 has a much greater relative abundance than mass
178.
According to published spectra, 2,4-dinitrotoluene does not have a
significant mass 121 ion, which does occur for 2,6-dinitrotoluene. We
suggest that the molecular ion (mass 182) be used for 2,4-dinitro-
toluene.
The relative retention time, as reported in Table IV, is obviously
incoi—3ct for bis(2-ch1oroethyl)ether which would be expected to
eluta- seora bis(2-cnloroisopropyl )ether. Also, the retention time
for tsoortorone is questionable. Retention times which we observed
on an -jV-17 column are attached.
All artarapts to chromatograph 1,2-diphenylhydrazine in our laboratory
resulted in a symmetrical peak giving spectra for azobenzene. In
addition, the standard was analyzed using the solids probe for
introcuction into the mass spectrometer and spectra were obtained
for both 1,2-diphenylhydrazine and azobenzene. The 1,2-diphenylhy-
drazfne did not show significant peaks at masses 93 or 105 as indicated
in Table IV. Azobenzane has significant ions at masses 77(100*),
105(dot), and 132(30:5).
The mass spectrum for N-Nitroso-di-n-propylamine does not have a
significant ion at mass 42. We suggest the use of mass 70 in its
place.
The Region VII Laboratory has had much better performance utilizing a
3% OV-17 column compared with the recommended 1» SP-2250 column. A
chromatogram demonstrating the separation attained on this column is
attached along with a listing of the relative retention times.
106
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Although we have been able to successfully chromatograph 40 nanograms
of benzidine, our laboratory cannot consistently achieve this recom-
mended nerformance level. We feel that this level is not practical
for a high-volume laboratory.
The recommended analytical procedure is not adequate for the differen-
tiation between certain isomeric pairs. These are anthracene and
phenanthrene, chrysene and benzo(a)anthracene, and benzo(b)fluoranthene
and benzo{k)fluoranthene.
Extraction Recoveries
No provisions are made in the procedure for determination of the
efficiency of the extraction process required for the base/neutral
compounds, phenols, and pesticides. We feel that this is an important
quality assurance technique which was overlooked. We have attached
some recovery values which are based on a very limited number of
runs. Note the zero recovery for hexachlorocyclopentadiene.
Phenols
Some of the relative retention times listed in Table V appear to
be incorrect for the Tenax GC column.
The Tanax columns (glass) develop rather large gaps over a period
of r;;n» and peak broacaning occurs. The packing material hardens
and t*?e columns cannot ba repacked.
Phenols analysis by the standard colorimetric method is a general
anal/sis for a long list of phenol compounds. The list of priority
pollutants has about eleven phenol compounds including phenol. Each
of these compounds are analyzed for by gas chromatography. Since
each compound is analyzed by GC, why analyze for a long list of phenol
compound by a nonspecific method? By dropping this analysis, we would
save about forty dollars per facility.
Data Storage
In order to minimize the amount of data storage on magnetic tape,
we suggest that nothing be stored for those GC runs showing no dis-
cemaole peaks. This would reduce the storage requirements by at
least 25*.
Radian Consent Decree Standards
Ethyl benzene is not present in the purgable compounds standard as was
indicated on the sheet supplied with the standard. Vinyl chloride,
2-chloroethylvinyl ether, and Bis(chloromethyl)ether are not present
in the mix. ^
107
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The data sheet does not indicate which isomer or isomers of 1,3-di-
chlorooropylene are present in the mixture. The 2-chloroethylvinyl
ether should be in the purgable standard rather than the base/neutral
extractables. It is too volatile (B.P. = 109°) to analyze as an
extractable compound.
The miscellaneous pesticides standard does not contain delta-BHC as
indicated on the data sheet.
The Arochlor mixtures and the Toxaphene/Chlordane mixture have very
little value in these analyses. The individual Arochlor formulations,
Toxapnene, and Chlordane should be provided as separate standards.
The crtQ-anthracene should be provided in a much more concentrated
(xlQO) solution. Ten micro!iters per sample is required utilizing the
2000 ??M standard. The phenols mixture and the base/neutral extrac-
tables standard should be provided with 20 PPM of d^g-anthracene
already included. This would facilitate determinatTon of relative
response ratios at one level.
We have been told that the concentrations for 4-Nitrophenol and
2,4-Oinitrophenol are incorrect as listed in the data sheet supplied
by Radian. The correct values are 50 PPM and 1GOO PPM, respectively.
FieTd Blank for Automatic Samplers
The snTy. interferring contaminants which we have observed in the
camccs-i-ar blank samples have been trace amounts of di-n-butylphthalate
and bis:2-ethylhex/1)pnthalate. However, these have never been found
at levels above our routine reported detection limit of 20 PP3.
Therefor*, we suggest that the number of compositor blank samples
be reduced to one per sampling site (per plant). Of course, a blank
shouts still be run TII Lhd Cib'TJ on every new batch of tubing which is
used.
Sample Size
The reccimiended sample size for the extractable organics is two and
one-half gallons. However, because of various field problems (batch
discharges, etc.) it is not always practical to provide that much sample
to the laboratory. Therefore, we ara recommending that two liters
be the minimum sample size which would be considered worthwhile to
even ship back to the laboratory. This would allow analysis for the
base/neutrals and the phenols.
108
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Analytical Time Requirements
We estimate that the total time required to do the 114 organic com-
pounds, 13 metals, and cyanide amounts to approximately four man-
days per sample with the bulk of the effort (ca. 3.5 man-days)
required for the organic compounds. This estimate does not include
sample collection. The limiting factor for the rate of analysis
(number of samples which can be analyzed per month) is GC/MS time.
The Region VII Laboratory has been utilizing two work shifts in
order -o derive maximum benefit from our existing instrumentation.
The SC/MS data output requirements with our present GC/MS configur-
ation anxDunts to about five hours per sample. Thus, under ideal
conditions (no instrumentation problems and no analytical problems)
the rnaximum output amounts to three samples per day (16 hours). This
number is further reduced to about two samples per day when one allows
for quality assurance procedures (running of standards, etc.). The
maximum rate for analysis of the priority pollutants is thus estimated
to be 40 samples per month with one GC/MS/Data system being used
16 hours per day with zero down-time. This rate could be improved
substantially for systems allowing data acquisition and data output
to be performed simultaneously.
Metals
Frcnr our experience only copper and zinc should be first analyzed by
flame stctuic absorption. Other metals are usually present at very
low concentrations and are analyzed for best by flameless atomic
absorption.
"Green List" of Priority Pollutants
Compound ?33 of the "Green List" is actually four different compounds -
cis-1,2-dichloropropylene, trans-1,2-dichloropropylene, cis-l,3-di-
chlorcpropene, and trans-1,3-dichloropropene. The analytical procedure,
however, does not refer to the 1,2-dichloropropylenes and it is not
provided in the Radian standards. We assume that the 1,2-dichloro-
propyTene on the :'Green List" is a misprint.
Other Changes
1. Dilution Water - Due to the possibilities for contamination in the
field, we will depart from the protocol requirement for a five liter
blank flush of compositor lines for each setup. Blank flush water will
be supplied by the laboratory in one-gallon containers. At the time
of setup, a single one-gallon container will be opened minimizing
exposure of the blank water to uncontrolled contamination. One and
I.,
109
-------
one-half liters of blank water will be flushed through the tubing
and disposed of and the ramaining two and one-half liters will be used
as a blank for evaluation of contamination from tubing. It is suggested
that the two and one-half liter portion be pumped from the one-gallon
glass holding and blank water to a one-gall on -glass jug emptied at the
previous station. This procedure will eliminate the need for funnels
and nrinimize opportunities for spillage and contamination.
2. Volatile Sample Containers - Handling of the volatile blanks
and collection bottles is critical. It is recommended that these
containers be refrigerated dry to the maximum extent possible including
transport to and from sample sites and holding spaces.
3. Phenol Samples - All phenol samples will be collected, preserved,
and handled according to routine Region VII procedures (one-liter
cubi, CuS04 and H3?04).
4. Cyanide Samples - All cyanide samples will be collected, preserved,
and handled according to routine Region VII procedures (one-liter
cubi, 70 pellets NaOH) with the following additions: (a) Litmus
paper -Trust be used to check for alkaline pH after preservation.
(b) Where chlori nation is practiced, the manipulation to zero residual
prior to preservation will be done using the Hach OPD kits to be
supplied by the laboratory and with ascorbic acid as the complaxing
agent according to the EGO protocol.
5. Three-Gallon Sample Bottle Tags - Field personnel will prepare
and artacn to the three-gallon containers, adequate tags for all analyses
for priority pollutants and BATEA parameters to be determined from the
comoosi-3. As a minimum, the following tags will be attached to the
composite bottle:
Manila - 8005, COD, pH, cond
Manila - NFS, (CI, S04, if needed)
Yellow - NH3-N, TKN, NC^-NC^-N, Total P
White - Do not list metals to be analyzed for, if it is covered
by the priority pollutant list.
Slue - Organics
6. Dissolved Parameters - In those few cases where dissolved para-
meters analyses are required, field personnel will be required to use
an aliquot from the three-gallon sample and provide field filtration
and preservation in separate containers.
1. Teflon Tubing - Teflon tubing is used for sample intake line
to the automatic compositor. The Teflon tubing is shipped back to
I.
110
-------
Region VII Laboratory, cleaned, and capped with aluminum foil before
being used at a different sample site. This has worked very well for
field operating procedures.
8. Data Handling - See attached memorandum dated October 6, 1977.
9. Instead of shipping blank volatile organic water samples to and
from tne sampling site, our laboratory will perform a weekly blank
check on glassware and distilled water.
Attachment
111
-------
1
UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
: October 6, 1977
: Data Handling and Reporting Procedures for Effluent Guidelines Division
Robert L. Jlirkey ^-
Director> Surveillance ;o/na/^AlTaC2Q data points per year and maximizes the utilization of
computer storage, collating, and retrieval of the data. This system
was developed in response to needs of the ilPOES program chain of
custody requirement and complies with the national program as out-
lined in the flPDES Compliance Sampling Manual (Exhibit 1). Complete
details ft.- normal application of this procedure are contained in
the Water Section Data-Handling System memorandum (Exhibit 2).
Much of the potential for dara turnaround is due to the development
of the local data system, Laboratory Management System (LAMS).
The LAMS data system is a Region VII system to accumulate and edit
data faaf;-» it is released to the user or to the Storet system.
The Labc-?:ory Management System is operational on the CQMMET IBM-
370/1S3 system. COMNET, in Washington, D.C., is the vendor for our
data sys:i~. The LAMS programs are stored on the vendor's USER and
WORK di;--;, and on Regionally dedicated 3330-11 disks. The Data
Processi"; Branch is responsible for the keypunching of all input
data. T-i 2ata Processing Branch inputs data with a Data 100 ter-
minal irr tr:? Regional Office. To input data to the system wa need
three reccrts: (1) a station location sheet that describes the
location :f t.-,a sample site in space, (2) a field sheet that assigns
a laboriusry number to information such as flow, sampling personnel
codes, szrnrling equipment codes, field measurement, and analyses
requester and (3) a laboratory bench card that gives the analytical
results fcr each sample.
After the analyses are completed, a LAMS Report printout is given to
the laboratory for editing. Aftar all corrections are made, the
laboratory releases the data to the user.
With the advent of the sample collection efforts for EGD, we have
encountered for the first tirne the need to accomodate confidentiality
of the data for certain samples. The addition of contractor labora-
tories participating in portions of the analytical effort also adds
a new aspect to our system. It is cur intent to identify key poten-
tial problem areas and recommended solutions for your review in this
\
112
EPA Form 1320 i (Re*. 3-74)
-------
submittal. In accordance with verbal directions received from
John fievorc'jgh on October 3, 1977, we will cease sampling activities
at specific process line effluents where confidentiality requests
from the discharger are expected until your staff has had an oppor-
tunity ts prepare a completely satisfactory progr&n based on this
package -»e ara submitting.
1. The field collection, handling, and shipment procedure under
normal c!~ai.n of custody is considered adequate and no procedural
modi fi carl errs need be made.prior to receipt of samples at the
regional laboratory.
2. Samples collected by the EPA field crews identify the individual
facility sa.-plad as well as specific sites at the facility in order
to provide assistance to laboratory personnel in establishing desir-
able levels for specific analyses and for comparing multiple samples
from a given site. An example of these tags is given below:
All sarnpies going to the contract laboratories are tagged and checked
through'CUT- crganics laboratory unit. Thesa samples are retagged by
laborator;' personnel with only the laboratory number and an industrial
code nuncar. As shown-below, the contract laboratory will only know
that they received a sample from Region VII from the petroleum industry
with the- lab number 321Q77.
S=S-i^-ir^S';i5s^--=.a-tS—^SCi^l r—Tt^-'^-f- ^.^ii.ii-J
^^g^o^^-^ra.—=--a -^-ss^rt-.|«.._^,^==^^rr,..,^
113
-------
Samples for contractor analyses are stored in a specific area away
from other samples and work in progress to prevent any access to
in-house samples by contractor personnel.
3. The regional laboratory facility is reasonably secure. Environ-
mental Protection Agency, SVAN parsonnel are the sole tenants; and
a four-.iijii combination lock entry system is provided to restrict
entry. Facility security could be improved significantly by more
frequen; changes of the combination and more restricted distribution
of the. access number outside of SVAN, Ideally, access to the build-
ing shculi be controlled by a contract guard/10 system on a 24-hour
per day basis. Such a system would provide a major improvement over
the alteT^cive choice which would require a multiple system of safes
and for secure file cabinets in at least six areas of the lab to
cover such items as field sheets, draft reports, custody sheets, bench
cards, 2£.rplas and data printouts, and a consequent major loss of
efficiency and increase in space commitment which could be used for
batter purposes.
4. As indicated earlier, the LAMS system is an in-house system
requiri" several access cades to reach the data and is a major
element •:.- .ur overall daca handling system. It is essential that
we mai.-t.i-~-> the LAMS sys~en in order to handle the volumes of data
in an effective manner. The potential confidentiality by utiliza-
tion of :i -is'jvord access- requirement by minor modifications in
the stev. •-• locator cards =nd by a more rigid control system on
transfer \~t release of ~.?e data output and elimination of the data
from the s'^nge system. These changes are being implemented and
v/ill res^l ~ in rigid documentation of the data management by the
SVAN Oat> Socrdinator and verified erasing of all results from LAMS
on a ti~»l/ basis after the final copy is approved.
5. For C--2- organic parameters, the first numerical data are pro-
duced whan chs organic chemist interprets the gas chromatogram and
mass scccrnl data. All of thesa numerical data are recorded on
one set c? summary sheets. The data coordinator v/ill keep these
sheets S3c-_re until they are transmitted to EGO by a chain-or-custody
sheet»
All mass spectrograph data are stared on magnetic disks. These
disks will be transmitted to Athens, Georgia, by chain-of-custody.
The Athens Laboratory will transfer these data to 9-track tapes and
erase the disks. The erased disks will then be returned to Region VII,
6. It is our suggestion that individual sample data from the con-
tractor analyzed samples be returned to the Region VI"! laboratory
for quality control review and compilation will all other samples
from a given facility in order to develop an easily understood
114
-------
single data package for each facility with an explanation of the
technical aspects of the field effort.
lection
7. All field records, notes, facility descriptions, and other_
information utilized by the field personnel for sample collectii
efforts ire held in a separate file within the SVAf-1 facility prior
to beirr. notified by EGO of the desired disposition of these items.
plete facility field data and the laboratory data package marked
confidential with all subsequent distribution to be handled by EGO.
This incl'j-:53 supplying NPOSS permit data to the appropriate EPA
Regional Office.
115
-------
UPDZS COMPLIANCE SAMPLING MANUAL
.. cBMsrrai. PHCTZCTZCT AGZSCY
OFFICE: OF WATZH
lie
-------
FOREWORD
The NPDES compliance inspection program represents a
significant commitment of resources by the States and EPA to
•the verification of permit effluent limitations and assurar.ee
that permit requirements for monitoring, reporting and
compliance schedules are being met and -enforced on a
nationally consistent basis. While compliance inspections
make up only one segment of the overall national water
enforcement prograa, they are highly visible and may be the
only direct contact that the permittee has with regulatory
perscr_r.el. Thus, compliance inspections must be performed
in a, thorough, professional manner, with nationally con-
sistarri coverage of key compliance elements. Reporting of
inspection data nmst also cover the key compliance elements
so thaz the data derived from this program can be aggregated
nationally, regionally and by States for purposes such as
program assesssact, budget development and reporting to
Congress. . . .
The previously distributed NPDES Compliance Evaluation
Inspection Manual CC2I) described the objectives and procedures
for carforming no a- samp ling inspections. The NPDES Compliance
Sajziplir.g Inspacticn Manual (CSI) describes technically sound
procedures, derived from the first hand experience of EPA
and Stat-e personnel directly involved in compliance inspections,
for tr.^ collection, of representative samples, flow measurement,
sartpl= handling . ar.d field quality assurance. .
The CEI and CSI 'Manuals and the revised Compliance
Insp action, Report: Fora, in conjunction with the annual
procr2= guidance ar.d other memoranda dealing with inspection
policy," fora ths frassawork. for the compliance inspection
progr3E2. Follo'^rLr.g tha procsduras and. policiss outlined in
thess documents will isprovs the quality of NPDES cotnpiianca
inspections , enhance the value of. data derived from these '.
inspections, and better serve the needs of the overall NPOES
enforcement program.
The manual is made-up in a loosa-leaf format so that
revisions or additions can be easily acccmodated. Any'
commer.-es or additions you may wish to make should ba
directed to the Compliance Inspection Manual Review
Committee, Compliance Branch (SN-338) , Enforcement Division,
Office of Water Enforcement, U.S. Environmental Protection
Agency, 401 M Street, S.W., Washington, B.C. 20460.
June 1977 Ass is tan y Administrator
for Enforcement
-------
SECTION VIII - CHAIN OF CUSTODY P3CC2DO-?SS
.-;- A. Introduction
As in any other activity that may be used to support
£.-/ litigation, regulatory agencies must be able to provide the chain
?:• - of possession and custody of any samples which are offered for
,- . - evidence- or which forst the basis of analytical test results
Z-Z-&T " introc^ad into evidence in any water pollution case. It is
~&~r'" iinperati-^2 that written procedures be available and followed
»_•'/ whenever evidence samples are collected, transferred, stored,
§x--c" •.
fV~; ' ' analyzed, cr -destroyed. The primary objective of these
*". •" *
K -
£-.".. procedures is to create an accurate written record which car: be
5 '-
«".— . ',
;: . used to trace the possession and handling of the sample from the
|- ' moment o- its collection through analysis and its introduction as
-j .-- evidence.
f. • • -.
*•-'• ^- ' ' ' •
|-V/: A sample is in someone's "custody" if:
py^'-. 1- It is in one's actual physical possession; or
M-t?-"-:' . 2. It is in one's view, after* being in one's physical. -
!fe---.- •
jgf^;-f--;.. • possession, or •
fCi/; 3. It is in one's physical possession and then locked up
5;V; • • so that no one can tamper with it; or • ._--.:.-
j.-. • ft. It is keot in a secured area, restricted to authorized
fl. - w
I ;' '. personnel only.
-113-
-------
B. survey Planning and Preparation
The evidence gathering portion of a survey should be
characterized by the conditions stipulated in the permit or the
miniciun number of samples required to give a fair representation
V.
of the ^astewater quality. The number of samples and sampling
locations, determined prior to the survey, must satisfy tha
requirements for NPDE5 monitoring or for establishing a civil or
criminal violation-
A copy of the s-ccdy plan should be distributed to all survey
participants in advance of tha s-urvey date. A. pre-surv«y
briefing is helpful to reappraise survey participants of the
objec-^iTss, sampling locations and chain of custody procedures
that vill be us»cL.
C. Sar-olinc Collection, Handling and Identification
1. T-T is. important tha-t a minisrca n-cs»ber of parsons be-
in ss^ple collecriiCTi and handling- Guidelines established in
this ra.nTial for saaipls- coHection, preaerva-feiart and handling
sho-cld. be used. Field records should be- ccmpleted at tha- tier*
ths sample is collected and should be signed or initialed,
including the date and time, fay the sample collector (s) . Field
records should contain tha following inf creation:
(a) unique sample or log number;
\— ,.
(b) date and tine;
-119-
-------
(c) source of sample (including name, location
. ; & sample type) ;
\: (d) preservative used;
;- pertinent field daca (pH, DO, Cl residual,
r "
f..-' etc.) ;
£-*,. .
-------
f
?r.j~
t" ^.
£'* -•
^
|->
2
f
o
•»
u
*
1
EPA,
S.'alian No.
S'aiioit < ~;aAvt*i
ann
S«ll,J,
can
Nuh4«^
Sarr.pl«r»
20 jSfiei SGJ»O< • iV * » A-. vmK-
D,>a
Maiala
DQ.
Ob
'
Tof. Coli*.
2 Ofeial Sfu*pi+
-------
-The use of the locked and sealed chests will eliminate the
need for close control of Individual sample containers.
Ecvever, there will undoubtedly be occasions when the use of
a chest is Inconvenient. On those occasions, the sampler
should place a seal around the cap of the' individual sample
container which would indicate tampering if removed.
£- When samples are composited over a time period,.
unsealed samples can be transferred from one crew to the next
crew. .-. list of sarrples will be made by the transferring crew
and signed for by a mejnber of the receiving crew. They will
either transfer the sasiplss to another crew or deliver them to
laboratory personnel who will then acknowledge receipt in a
similar -arner. • - -. ."-"•
5-_. Color slides or phonographs taken of the
"outfall location and of any visible pollution are recaurm-nded to
facilitate identification and later recollection by the "-.-.:
Inspec-rcrr, A photograph log* shotzld be inade at the tictsx tha photo
" • . '
is taker: so that this information can be %-ritren later on the-
back of -he photo or the margin of the slide. This shonld .-
include the signature, of tha photographer, tine, date, site
location and brief description of the subject of the photo.
Photographs and written records, which nay be used as evidence,
should bs handled In such a way that chain of custody can be
established, •--..-
-122-
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D. Transfer of_ Custody and Shipment
1. when tranaferring the possession of 'the samples, the
transferee must sign and record the date and time on the chain of
custody record..- Custody transfers, if -tnade to a sample custodian
in the field/' should accoxrrvt for each individual sanple, although
samples may be transferred as a group. Every person who takes
custody must, fill in tha appropriate section of the chain of
Custody F.ecord, To prevsrtlr undue proliferation of custody
records', the number of custodians in the chain of possession
should be as few as possible. •
.'2. The field custodian or field inspector, if a custodian
has not been assigned, is responsible for properly packaging and
disca-^iing samples to the appropriate laboratory for analysis.
This responsibility includes filling out, dating, and signing the
approcriate" portion-of the Chain of Custody Record. A Chain of-
Custody Record forriac containing tha necessary procedural, elenent
is illustrated in Figure VIZ1-2. . • "'"••"•. ~."'•'.;
packages- sent, to the laboratory should be
accompanied by the Chain of Custody Record arid ether pertinent -
forms, a. copy of these forms'should be retained by the- . _ " ;_
originating office (either'carbon or photo copy).
ft. Mailed packages can be registered with return receipt
requested. If packages are sent by common carrier, receipts
-123^ •
-------
FIGURE VIII-2
CHAIN O? CUSTODY RECORD
. -A
SURVEY
i
iru"3°* I irArtOH iocj-:f°'4
i
[
•
OAf?
•" .
Relinquish--: ryr ^r,.,,,..^..^
Ralinquiarf-s-j ry: ."> .*. .y
Relinquishes by: rs&m**^
RsJinqs/t3rt«j by: fs^
Dispc^ch^a by: (s^^*^
Meffiod of Shipman^:
TlMe
SA.M?tE.RS;/s-ji... .j
SJ..M*t= rv?«
W^.T,
C«-p.
Gf3*.
sza.
NO.
-
NO. Or
CS^fAtMr*!
ANAirsii
*«QU'*rO
•
.
Recff**^ b^r is-j. -><. ••) . .
Rscstve*i byr fTTj...i..ij
Received by: fs-j. „.»-.>
Hece»v-c by Mobii* Lcbora,vory /or ?i^ .
Dat-/T;m»-
' 'I '
Daf«/T;,-n--
• i
Dafs/Ti/rr*
Dcfs/Time
• 1
Daf?/Tim»
I
I Copy—-Sorvwy
-124-
-------
-
--
should be retained as part of the permanent chain-of custody
docuraefita tion.
5. samples to be- shipped must be^so packed as not to
break and the package'so .sealed or locked that any evidence of
tarnperir-g may be readily defected-
v~~-—,
Jisjf-Ss
rahcratorr Custod? Prrscgdsnsa
~ "" - '
?n of .Custcxrr procedtzrea are also necessary in the
^•-^_7 fcon -the t±s» of sainpla receipt to the ticre- the sample
is discarded. The following procedxirea are recommended for the
laboratory?
1.
absence,
A specific perscrr shall be designated custodian and an
.« designated to act- as custodian in the eaatodian-s-
all inccoing sactplea shall be received by th*\_
,'vber shall "indicate re^eirri by signing the accc^oanying
custcdT f oms aBid'^bo'shall irtaia _th»-signed fo==o
records. -' . .-^.^v - ' ""- • -' '
• 2- The sasiple
book ta record, - for- each
sample, the per
source of sample,
transmitted to the laboratory
sh
:tai-n a pensan-errfc log
*Z*~°* a«li«ring tii.
dcn cr log nu^er,
concision received (sealed,
-125-
-------
unsealed, broken container, or other pertinent remarks) , A
standardized format should be established for log'b'ook entries.
3. A clean, dry, isolated roocn, building, and/or
refrigerated space that can be securely locked fro:n the outside-
shall be designated as a "sample storage security area."
' '4. The custodian shall ensure that heat-sensitive, light-
sensitive samples, radioactive, or other sample materials having
''unusual physical characteristics, or requiring special handling, .
are properly stored and maintained prior to analysis-
5, Distribution of samples to the section chiefs who are
responsible for the laboratory performing the analyses shall be
inade only by the custodian. • . •-*..'••.:-•.•••_
5. " The laboratory area shall be maintained as a. secnrsd •
area, restricted to authorized personnel only. .-*'.--•
7. " "Laboratory personnel are responsible for the cars-and "
custody- cf the sample- once it is received by them and shall be
prepared co testify that"the sample was in thair possession:and
view or secured in'the laboratory at all tirnes from the mornervt: at
was received from the custodian until the time that the analyses
are comoleted. ' ' r • •
\
-126-
-------
.
£/;'-
r-r;t-;
»-• /*•"*
v __ •. *
8. Once the sample analyses are completed, the unused
portion of the sample, together with all. identifying labels, must
be returned to the custodian. The returned tagged sample should
be retained in the custody roots until permission to destroy the •
• ""•
sample 1 3 received by ths custodian. - . .". -
5. Samples shall be destroyed only upon the order- of the
•'
Laboratory Director-, in consultation with previous iv designated -_;
• • "•••.-••--
Snforcerrerrt officials,, or wh«n it is certain that the inforaatiorr
' ••'-•.
is no longer reqtsired or the samples have deteriorated. . The s
"
• ,-.-
procedtire is. true for tags and laboratory records. . .--:?":
F. 'ErrL z errtlarv Cor:si.dgratioTT3 - . • :.
.- •• Heczicing c^J-r. of custody procedures as well as the-' various —
promulrated laboratory analytical procsd-crss to writing will -.-. ._
facilitate tha adnlssiorj of evidanea- rmder rule 803 (S) or. tfta- .'..^
sZ. Rules-, of z-riderjce (?L- 93-575} .- • Under- this
written records of regrslarly . conducted busines9 - activi-tie»-r-inay -'^1
.'-...- ^ • • • •'• .'-y'-Tr.y--^--"-^?
be irrtrocuced- into- evi degas as art e-ggagt-.cn to tfc« "Hearsay.- Hole*-
withotrt the testictony of the- person i";i-J— r-Jrr
":^'.-.~-i~-1-v-":='5^
individuals -who collected, 'kept:,, and analyzed samples testify .-in-:
court.--- In addition, if the opposing party does not inteni.to--.r-i
.>n>. •:.-
contest the integrity of the_ sample- or. testing evidsnct-s,. •'Ir r :. , .."
adsiission under the Rule 803(6) can save a great deal . of -.trial . ,
^ '
time. For these reasons, it is important that the- procedcrss
-127-
-------
followed in the collection and analysis of evidentiary samples be
standardized and described in an instruction manual which, if
need be, can be offered as evidence of the "regularly conducted
business activity" followed by the lab or, office in generating
***
any gi^-'e^ record.
In. criminal cases however, records and reports of matters
observed by police- officers and other' law enforcement personnel
• * " .
are not included under the business record exceptions to the
"Hearsay ?.ule'T previously cited (see Rule 803(3), P.L. 93-595).
It is arguable that those portions of the compliance inspection
report dealing with matters other than sampling and analysis
results come within this exception. For this reason, in criminal
actions records and reports of matter observed by field
investigators may r.ot be admissible and the evidence may still
have to.-be- presented in. the form of oral testimony by the
person (s* who made the record or report, even though the • '. '•".
materials ccnre-. within the definition of business records. -In a
criminal oroceedirig, the opposing counsel may be abla to obtain.
copies- of reports prepared by witnesses, even if the witness does
not refer to-the records while testifying, and if obtained, the. .
records nay be used for cross-examination purposes.
Admission of records is not automatic under either of these
sections. The business records section authorizes admission
"unless the source of information or the method or circumstances
-123-
-------
of preparation indicate lack of trustworthiness," and the caveat
under the public records exception reads "unless"the sources of
information or other circumstances indicate lack of
trustworthiness, "
Thus, whether- or not the inspector a./ " -ipates that.his or
her compliance inspe:-..* • he or slia- should make- certain that the report is as accurate an
1-? - -" ~ •
•>:"•.•:."•'•-.. .objective as possible-- ' -'
-129-
-------
C.Q_
Green List
Number
Compound Name
i
-.24
2-Chlorophenol
• 65
'. Phenol
31 ,
2 ,4-Dichlorophenol
57
• 2-Nitrophenol
. 22
P-ch 1 oro-M-creso 1
21
2,4,5-Trichlorop-s— : 1
34
2, 4- Dimethyl phenol
• 59
2,4-Oinitrophenol
60
4,5-Dim'tro-O-crescl
58
4-Nitrophenol
64
Pentachlorophenol
: ^EKE**
**"
STORET
Number
34586
32730
34601
34591
34452
34621
34606
34616
34657
34646
39032
—
Sample Number
23'^3«-»
3 /•x.l"'
3*
^ .
S*^ " 7
w
130
]U(c
'*
]
•ZG3
nj
«c
(jiiiJ
-t.
Q.II
C^.53
1 C^
1,00
I.3M
/.a
^
13s*
OS"
.^z
1 ^ 7
*$
•z.ca
|1Z
;s
i?v
(•3
77
(p5
13?
2
-------
|cc/,aci
Green List
Compound Name
26
1 ,3-Oichlorobenzene
27
1 ,4-Dichlorobenzene
12
i
Hexachloroe thane
25
1 ,2-Dichlorobenzene
42
Sis (2-chloro isoirooyl
ether
52
Hexachloro butoilars
3
1,2,4-Trichlorobs.izsne
55
Napthalene
18
Sis (2-chloroethyl ether
53
Hexach 1 orocyc 1 o-
pentadiene
56
Nitrobenzene
.43
Sis (2-chloroethoxy)
methane
20
2-Chloronapthalene
STORE!
Number
34566
34571
.
34396
34535
34233
34391
34551
39250
34273
34386
34447
34278
34200
v 1
Sample Number
^O^Kroius
fVU,irv<*'3l
"3 /V,!-'
II
is"
13
0.1.
T? "~"\
3 VJ
s-7
fel
(a?
f\ /-)
^S
^
/_5
HI
'
RcC»tj.vC
T*
HCB
Q.Ml
o.qa
&i
Q.MO
__ / ( ^
U- M 4
a. s^
Q.UC
Q.(oa
,
c'
Q.7C
a.r7
Q. (P 3*
0.7?
U^^TQ
IMS
H3
IM<*
113
in
ZC,
mu
113
MS"
"77
oatr
-2.-2.-7
7^
/0
-------
PRIORITY POLLUTANTS: Base/Neutral Extractables
Page 2
Green List
Nuraber
Compound Name
54
Isophorone
80
Fluorene
35
2,5-0-initrotolue.ie
37 / \
l,2-0iphenyl-hydr22in9
3S
2,4-Oinitrotoluer:s
62
N-Nitrosodiphenyi jST-e
9
Hexachlorobenzene
41
4-3romo?h«nyl phervl
ether
31
Phen»nthrene
78
Anthracene
71
Oimetnylphthalate
70
Olethylphthalate
39
Fluoranthene
STORET
Number
34408
34381
34626
34346
34611
34433
39700
34636
34461
34220
34341
34336
34376
S4
Sample Number
—
£
/ s" W
i ijij
0,o
,tr
j [g (f
ni
»7J>
193
.«
IS'J
/u
234,
2Vt
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Q.9C
(0.9 s-;
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lol
,c
S7
w
^
9,
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1 o(<
9-7
9^
«•<•
"
l«
no
,
-
Pyrene
34469
;^^fisas»ae5g*
>.'&«&nftJu£riCAr>
-------
PRIORITY POLLUTANTS: Sase/Seutral Extractable*
Green List
Number
Compound Name
63
Oi-n-butyl-phthalsts
5
Senzidine
57
Suty 1-benzy 1 -phtfta : its
76
Chrysene
65
Bis(2-ethylhexyl)
phthaJate
72
Benzo(a)Anthrscars
74
8enzo(b)Fluoranth«re
75
8enzo(k)Fluoranthana
73
Senzo(a)Pyrene
33
Indeno( 1 ,2 ,3-cd ) -pyrene
82
Oibenzo( a, h) -Anthracene
79
3enzo(g,h,i) perylene
fil ,
H-N i trosod i methyl ami ne
STORET
Number
39110
39120
34292
34320
39100
34525
34230
34242
34247
34403
34556
34521
34438
Sample Number
S3- n*.
a/4
253
^1
CS7
W
a?7
3C.7
ij \t *
fi Q ^
"T
W
«r
...
MS
I.BO-
|.?8
i.^
,../
''^
|.7Q
/ 70
u?
Q'^
*«
I
x«
mi
,OM
tw
.U
1*41
-------
PRIORITY PCLLb'TA?!TS: Base/Neutral Extrsctables
Green List
Number
Compound Haas
40
4-Chloro-phenyl p^eryl
ether
28
3 ^'-Oictilorobenz-iiirie
69 .
V
Oi-n-octyl pntalcta
77
Acenaphthylene
°*££££,,f
STORET
Number
34641
34631
34596
34JOO
, »• » -i
Sample f.'umber
,,,
ISM
,«s
^
1 30
,«
•
^r
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,.«
0.^
,.^
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IM1
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pocr-°°
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i
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' 4
f
•sf^.-.f ' •••£•;
134
I -.— '.^^T:
» ^,
1
-------
PRIORITY POLLUTANTS: Volatile Organic*
Green List
Number
Confound Nasie
2
Acrolein
3
Acrylonitrile
45
Chlorome thane
50
Qichlorodifluoromethane
45
Bromonethane
38
Vinyl Chloride
16
Chloroethane
44
Methylene Chloride
49
Trichlorofluoromat.line
23
1 , 1 -0 i cii 1 oroethy 1 ene
30
Trans-1 ,2-Oichloro-
ethylene
23
Chloroform
10
1, 2-0 1 Chloroethane
STORET
Number
34210
342TS
34418
32105
34413
39175
34311
344Z3
34438
34501
34546
32106
34531
11 1
1,1,1-Tnchloroethane 34505
V^JUIJJLU.I, '
Sample Number 's C1-
SS^TJN-A
(VUlx^etV
—
—
—
—
1
IS"
11
OS"
2C
33"
S3
74
S3
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^
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i'_«.a»sS8SS
H
;
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'- --X-v' V;*V < >7l-:y^v;-^illf
?i^;'£j^'i>af^&>V<<-''^^=^
-------
PRIORITY POLLUTANTS: Volatile Orgamcs
f
2.:
I-.- .
• *»-*s--
Green List
Compound Name
6
Carbon tetrachlorida
48
Bromodichlorone thane
17
Sis-chloromethy] ethsr
32
1,2-Oiciiloropropane
33 A
Trans-1 ,3-Oichlor3-
propene
51
D i bromoch 1 ortj-st -i-s
33 3
C-is-1,3-Oicnloroprojene
14
1 ,1 ,2-TMch Joroetnane
4
Benzene
19
2-Chloroethylvinyl ether
47
Sronwfortn
15
1 ,1,2,2-Tetrachloroethan
35
1,1,2,2-Tetrachloroether
STORET
Number
32102
32101
34268
34541
34561
34306
34561
345 n
34030
3457S
32104
34475
34516
13 !
Sample Number
O ^
l Q
in
,*.
133
isa
'**
* r
5=1
1 3^
,5,
in
205
. a n
o-s,
atlM
Q.U1
Q.13
G-S3
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1.0-1
1-14
i ii"?
'
wr.
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.37,
-------
PRIORITY POLLUTANTS: Volatile Orgamcs
Page 3
Green List
Number
Compound Name
Z7
Trichloroethylene
86
Toluene
7
Chlorobenzene
38
Ethyl benzene
<&&Qf>vi ^W(.C JIG
P-|
i"3
QQ7
•
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0.73
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1.34
o.a^
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97
I3C
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130
l 13
s-s-
^c
93.
•
j
• i
-------
PRIORITY POLLUTANTS: Pesticides
Green List
Number
Compound Naine
96
a-endosulfan
'02
o-SHC
!04
Y-8HC
(03
3-3HC
39
Aldrin
iCO
Hsptachlor
'01
Heptachlor epoxiss
95
a-sndosulfan
93
Oieldn'n
93
4, 4' -ODE
94
4, 4 '-000
92
4,4'-OOT
93
EndHn
STORET
Number
34356
39337
34264
39333
39330
39410
39420
34361
393SO
39320
39310
39300
39390
97 1
San"p'e Number
ql^SG-iQ
(Son-
TO
,.r.
*.«
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C.fl
l.oo
o.rz
1.37
,.-»
2,'t
1.1*
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-TO
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138
-------
Environmental Research Laboratory
UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
„„_,„ Athens, Georgia 30605
DATE: January 18, 1978
SUBJECT;Action Concerning Region VII Comments on "Sampling and
Analysis Procedure for Screening of Industrial Effluents for
Priority Pollutants"
F«OM:W. M. Shackelford
Analytical Chemistry Branch
TO: William A. Telliard
Environmental Protection Agency
Effluent Guidelines Division
WH-552, 401 M Street, SW
Washington, DC 20460
In a memo dated 2 November chemists at the Region VII
Surveillance and Analysis Laboratory commented on strengths
and weaknesses of the analysis protocol for the consent
decree analysis program. Several of the "deficiencies and
errors" mentioned in the Region VII memo refer to typographical
errors, while others are the result of differences in judgment.
This memo will deal with each comment from the consecutive
sections "Base/Neutral Compounds"—"Radian Consent Decree
Standards" in the Region VII memo.
Base/Neutral Compounds
A) The protocol should be corrected such that 177 is one
of the characteristic masses for diethylphthalate
instead of 178.
B) After reflecting on the ions for 2,6-dinitrotoluene and
2,4-dinitrotoluene, it appears that corrections should
be made such that:
2,4-dinitrotoluene—165(100) , 63(31), 182(11)
Apparently, the ions for 2,6-dinitrotoluene were put in
the space for 2,4-dinitrotoluene by mistake.
C) It is true that bis(2-chloroethyl ether) elutes before
bis(2-chloroisopropyl ether) on 3% OV-17 but this order
is reversed on 1% SP-2250. We observed this in our lab
as well.
D) Published values for the 42 ion for N-nitroso-di-n-
propylamine all indicate a significant intensity. Mass
42 is also characteristic of all alkyl nitrosamines.
E) The diphenylhydrazine problems have been commented upon
in a memo to you from Wayne Garrison and Fred Haeberer.
EPA FORM 1320-6 (REV. 3-7S)
-------
-2-
F) 3% OV-17 was tried in this lab and found to give bleed
problems. The 1% SP-22SO is equivalent to 1% OV-17.
Our work showed essentially equivalent chromatography
with the two packings (3% OV-17 and 1% SP-2250) except
for some changes in retention times .
G) Chromatography of benzidine is a necessary evil, but
the amount used could be increased to 100 ng.
H) We are aware of several PAH isomers that cannot be
separated on the recommended column. They cannot be
separated easily on capillary columns either. We will
have to live with reporting them as the sum of the two
isomers .
Extraction Recoveries
*.
A) A provision for the determination of extraction recovery
efficiencies should definitely be made for the verifica-
tion stage. Other labs have not experienced troubles
extracting hexachlorocyclopentadiene. This probably
needs study .
Phenols
A)
Relative retention ti-T.es fcr phenols in the protocol
should be corrected as listed below:
RHT
Phenol 0.79
2-chlorophenol 0.34
2-nitropher.oi 1.00
2, 4-dime-chylphencl 1.02
2, 4-dichlorophenoi 1.06
p-chloro-ni-crasol 1.27
2 , 4 , S--crich.lcrophe.rioi 1.34
2,4-ci2itrcphenci 1.53
. 4-nitrophenol 1.73
4, 6-dinicro-o-cresol 1.32
pentachlorophenol 2.01
These agree reasonably well with other data. The
original data was taken before Tenax GC had been fully
evaluated in this lab.
3) One help for t£e gaps in Tenax is to condition a new
column at <_225 C, pack together when spaces develop,
then use normally. This can minimize the gap forma-
tion.
140
-------
-3-
C) You have commented previously on the use of the 4AAP
method for phenols.
Data Storage
A) It has been stressed that each VOA, B/N, and Acid
extract must be run in the GC/MS and the data saved.
, In following up this point with Dr. Kleopfer, he
assured me that he is running all the samples on the
GC/MS and not just screening with GC to avoid GC/MS
samples with no flame detected peaks.
Radian Consent Decree Standards
A) Radian inadvertantly left ethylbenzene out of the VOA
mix. Vinyl chloride and bis(chloromethyl ether) are
present in the mix. Dr. Tom Bellar has commented that
the vials must be opened at 20 C to keep from losing
these two. The 2-chloroethyl vinyl ether was put in
the B/N vial. New standards have been promised by Dr.
Larry Keith of Radian. He has been made aware of the
shortcomings of the first set. The new sets will have
more divisions for more ease of identification of
individual components. Concentrated samples of the
d, ,,-anthracene have been on order for several months.
Dr. Keith said that contractual problems in HQ were the
hold up.
3) Corrections to the standards identification sheets have
been made to reflect the proper concentrations.
141
-------
THIS PAGE LEFT BLANK
INTENTIONALLY
142
-------
THIS PAGE LEFT BLANK
INTENTIONALLY
143
-------
UNCHLORINATED BASE/NEUTRAL PRIORITY POLLUTANTS
ORDER OF ELUTION
Protocol (EPA)
Identification by
Gas Chromatography
(Radian)
Compound RRT
naphthalene 0.57
acenaphthylene 0.83
acenaphthene 0.86
isophorone 0.87
fluorene 0.91
phenanthrene 1.09
anthracene 1.09
dimethyl phthalate 1.10
diethyl phthalate 1.15
fluoranthene 1.23
pyrene 1.30
di-n-butyl phthalate 1.31
butyl benzyl phthalate 1.46
chrysene 1.46
bis(2-ethylhexyl) phthalate 1.50
benzo(a)anthracene 1.54
benzo(b)fluoranthene 1.66
benzo(k)fluoranthene 1.66
benzo(a)pyrene 1.73
indeno(l,2,3-c,d)pyrerre 2.07
dibenzo(a,h)anthracene 2.12
benzo(ghi)perylene 2.12
Compound RRT
isophorone 0.46
naphthalene 0.51
acenaphthalene 0.81
acenaphthene 0.83
dimethyl phthalate 0.88
fluorene 0.92
diethyl phthalate 0.98
phenanthrene 1.10
anthracene 1.10
dibutyl phthalate 1.23
fluoranthene 1.30
pyrene 1.34
butyl benzyl phthalate 1.51
benzo(a)anthracene 1.57
bis(2-ethylhexyl) phthalate 1.57
chrysene 1.57
benzo(b)fluoranthene 1.74
benzo(k)fluoranthene 1.77
benzo(a)pyrene 1.80
indeno(l,2,3-c,d)pyrene 2.07
dibenzo(a,h)anthracene 2.13
benzo(ghi)perylene 2.13
144
-------
Calspah
19 December 1977
PMT:hf-67
Mr. William Telliard
Chief, Energy and Mining Branch
Effluent Guidelines Division (WH-5S2)
USEPA
Washington, DC 20460
Dear Mr. Telliard:
This letter is for the purpose of updating you on the situation
regarding the transport of hazardous materials which was mentioned at the
Denver analytical seminar. As you are probably aware, the Department of
Transportation has a regulation (49CFR172-101 Hazardous Materials Table)
forbidding the transport of nitric acid aboard passenger aircraft. In
keeping with the requirements of the regulation and in view of the fact
that the EPA Standard Method for metal analyses calls for acid stabiliza-
tion of samples to pH <2, Calspan filed for an exemption to this regulation
so that field sampling for LOE Task 11 would proceed uninterrupted.
On December 15, 1977, Calspan received a reply from the DOT
denying our request to transport nitric acid (MOO ml) in a specially
prepared field sampling kit. (Enclosed is a copy of Calspan's request
for exemption describing the conditions under which the acid shipment
would take place and also a copy of the denial.)
Since the most recent revision of the sampling protocol specifies
field stabilization of metal samples (verbally given by you at the Denver
seminar) and in view of the recent ruling by DOT on our request, we feel
that this situation should be brought to the attention of all contractors
involved in the field sampling phase of the Effluent Guidelines Program.
This regulation by the DOT may seriously jeopardize the ability of con-
tractors to provide accurate metal analyses on unstabilized wastewater
samples. We would appreciate any assistance on your behalf to resolve
this situation with DOT and kindly request you inform us of any change
which may be affected by your action. Thank you.
Sincerely,
P. Michel Terlecky, Jr. Ph.D. ' (
Head, Environmental Sciences Section
Environmental § Energy Systems Dept.
Enclosures
-------
Caispan
PC. Box23i
Buffalo, New York14 221
Tel. 1716j 632-7500
28 September 1977
PMT:pL-37
Office of Hazardous Material Operations
U.S. Department of Transportation
Washington, D.C. 20590
Attn: Exemptions Branch
Gentlemen:
Item 1.
In accordance with subpart B, Section 107.103, Caispan Corporation
seeks exemption of the requirements of 49CFR 172.101 Hazardous Material
Table (HNO, forbidden aboard passenger aircraft) and seeks to determine
what may be carried in "Chemical reagent kits" as described in Section
173.286. According to Section 107,103,b(l), three copies of this request
are submitted herein for your review and approval.
It em 2.
Specifically, we seek to carry aboard passenger aircraft chemical
reagent test kits during sampling expeditions in support of requirements of
the U.S. Environmental Protection Agency (USEPA Contract 68-01-3281) to set
national effluent standards for various point source categories pursuant to
P.L. 92-500 (Federal Water Pollution Control Act Amendments - 1972) and
various state, local, and regional agencies and industrial customers. The
chemical reagent test kits are necessary in order to properly preserve
wastewater samples for subsequent analysis in our laboratories in Buffalo,
New York. Without certain stabilizing agents, these samples degrade
resulting in a loss of value of the sample for analytical and regulatory
purposes.
Item 3.
The applicant for this exemption is:
Caispan Corporation
A, tn: Environmental and Energy Systems Department
P.O. Box 235
Buffalo, New York 14221
716-632-7500
146
-------
Item 4.
In accordance with DOT 15(A) spec. 173.263(a); d(l) and i(l), the
proposed -nejthod of shipping is as follows: a small plastic container (bottle)
with a threaded acid-resistant cap cushioned by absorbent packing material
is enclosed in a glass bottle with threaded acid-resistant plastic cap. This
glass container is then enclosed in an individual, tightly sealed metal can
and surrounded by vermiculite (mineral matter) packing inside. The metal
cans are then placed in wooden boxes, surrounded by cushioning material
(vermiculite). The wooden boxes are then secured with lids, screwed into
place and properly labeled as to the items contained therein.
The wooden boxes mentioned above are constructed of white pine
stock with 3/4" walls and have reinforced ends with a total thickness of
1 1/2". The lid is also constructed of 3/4" stock and when secured in place
with 1 1/4" screws affords an effective seal capable of withstanding trans-
portation handling. Photographs of this proposed method of shipping are
attached. Construction blueprints were not utilized in the assembly of this
item and hence are not included in this report.
Drop tests have been performed on the proposed transport containers
(wooden boxes) in accordance with DOT 15(A) spec. 178.168-6: Gluing Efficiency
Wood Drop Test. The specification states that for containers with a gross
weight less than one hundred and fifty (150) pounds, the container, when filled
to capacity with sand and/or sawdust, shall be capable of withstanding eight (S)
drops from 1 foot (12 inches) onto solid concrete, 1 on each corner, without
exposure of contents. Such performance has been attained (and exceeded) on
the proposed containers and it is expected that these containers will withstand
the type of handling normally associated with transportation of materials on
commercial carriers.
Item S.
See attached table.
Item 6.
We believe that the level of safety achieved will meet or exceed
that required by the regulations and will ensure that no additional risks
to life or property will occur as a result of the granting of this exemp-
tion. Our record of shipment of explosives and other materials over the
past 30 years is exemplary with four full time personnel at Calspan devoted
to packaging.- shipping and receiving activities alone. Because of our
experience, and because the small amounts of reagents needed present no
additional risk, we respectfully request approval of our exemption at the
earliest possible time.
147
-------
Item 7.
The proposed mode of transportation for the chemical kits is
by commercial aircraft. Separate shipment of chemical reagent kits by
other carriers or by cargo aircraft will seriously affect our ability
to support EPA requirements in rural areas of the U.S. and would adversely
affect our acquisition of some $1 million worth of environmental sampling
and analytical business annually. Receipt by our engineers and technicians
of these kits simultaneously with receipt of sampling equipment and con-
tainers (shipped as baggage) is essential to the timely and economical
conduct of our work for which the Federal government is the main supporter,
Due to the small amounts of material being shipped (3 containers
of 100 ml (3 oz.) each) and considering the extraordinary care under which
these kits are prepared and packaged, it is felt that there will be np_
increased risks associated with the shipment of these materials.
Item 8.
As previously mentioned in Item 2, these kits are required for
the proper execution of the water sampling phase of a variety of EPA-
sponsored programs directed toward the establishment of national effluent
guideline regulations for numerous point source categories. This work as
described by P.L. 92-500 is a continuous effort and is due to be continued
for an indefinite period of time. Each sampling trip arranged for data
collection in conjunction with the above-mentioned programs lasts an
average of 4 days. During this time the chemical kits are transported to
the sampling site, used as required and when empty, returned to Calspan.
Item 9.
The small amounts of materials due to be transported do not
constitute any increased safety hazard when packaged and shipped as
proposed above. It is strongly felt that the time and effort invested
in the execution of precautionary measures for shipment of these hazardous
substances is consistent with the public interest and will adequately
protect against the risks of life ana property which are generally asso-
ciated with the transportation of hazardous materials.
Item 10.
It is not necessary to process this application on a priority
basis. We do ^espectfully request, however, that the handling of this
resubnitted application be given all due consideration with regard to
expeditious review. This is necessary in order that our company may not
experi -,ze any interruption in the acquisition of the aforementioned
govern, .;nt contracts which represent a significant financial investment.
148
-------
If there are any technical questions related to the materials
to be carried or the kits themselves, contact our Dr. P. Michael Ter.1 eoky
at (716) 632-75CC, x538.
Sincerely,
/
Poland J. Pilie, Head
Environmental § Energy Systems Department
149
-------
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151
-------
\ /'' i'\ \ DEPARTMENT Or TRANSPORTATION "^P
I f,' / ' -, •]: A\ATER1AL5 TRANsPOkTAl'ON BUREAU
^V- " ' ""V^ WASHINGTON, O.C. IOS9O
i
| . DEC S 1377
I iSr . Xoland 3 , Pilie
1 Environmental and Energy
) Systems Department
.CaJLspan Corporation
•j P.O. Sax. 235
•I Boffalo, ItewTork 14221
j
1 Jleax Mr, Pilie:
i
| This is in response to your application dated September 2B. 1977
| (7844-N), iiled in accordance with Section 107. 103 of Title 49, Code of
i .Federal Regulations, (49 CPU), for -permission to snip cnemical kits
containing 70 percent .nitric acid, .sulfuric acid, -pnosphorii; acid ana
..solid .sodium iivdroxide pellets by passenger, carrying aircraft.
i
^ -3si accordance -with the Code of Jedsral- Regulations, Title 49, Section
\ . 107^ 109(0, .the request -is denied.
^ . The TEason/T-or denial is failure of -tne application TO saxisxy
\ requirements of Section 107.103 as follows:
] 1. In accordance -with 49 CFR 107. 103 Cb) (9) (±) you have made general
3 .statements as to why you believe that your, proposal to include 7H^
I ; nitric acid in 'the chemical kit -will scnxevt a level oi satet? at. iaa-_-
equivalent to that specified in the regulation from which the exempt ici
* is sought, .^owevex, it .is .obvious -that ,-n.o. ;yractT cal pactogiog ^tor &&•••
4 hazardous material will result in the same level of safety as -will be
| achieved by -precluding that material from transport.
j . 2~ Jlso. 49 CTS 173.286(b) and (n) (1) limits the contents of chemical
j .kits to 'corrosive liquids for vhicb B3^eption5 are provided in 49 CFR
i 172,101. Therrexore, nitric acid 01 caacentTanon of 4ui. OT less is n. ^
\ anthorized to be iticluded in a chemical kit. It. 'would, theraf ore, bvj
' more hazardous to permit nitric acid of concentration exceeding % ro
4 ie .included in a. chemical . kit .thereby further reducinp. the specif ieil
1 level of safety.
152
-------
idition as -noted below., -same of your requests axe unnecessary in
that the regulations already .authorize .shipment by passenger carrying
il-craft.
s tion T73 286 (b) provides ior shipioent cf sulfuric acid and -phosphoric
-• ' in .chemical .kit?. under conditions, that jaake your request for exemption
f<-r" these commodities unnecessary.
-tion 173 744 provides ior -shipment of sodium hydroxide solid as a
Intc-d quantity"such that no exemption is necessary for the package you
, -..rribi-a in your application .
Sincerely ,
Alan I. Roberts
Director
Office of Hazardous -Materials
Operations
153
-------
UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
ENVIRONMENTAL RESEARCH LABORATORY
ATHENS. GEORGIA 30601
December 22, 1977
Mr. J. B. Anderson, Editor
Analytical Quality Control Newsletter
EMSL
U.S. Environmental Protection Agency
Cincinnati, Ohio 45268
Dear Mr. Anderson:
Enclosed is an article describing preliminary results of
investigations by Dr. Fred Haeberer of the Analytical
Chemistry Branch into the stability of two of the Consent
Decree Pollutants. This should be of interest to the many
of your readers who are involved in analysis of these
pollutants, and we request that you publish it in your
Newsletter. We are also planning to publish these results
in the Athens ERL quarterly report, which will probably be
published in 2 or 3 months.
Sincerely yours,
Arthur W. Garrison
Analytical Chemistry Branch
cc: Dr. Jim Lichtenberg, EMSL, Cincinnati
Dr. William Telliard, Effluent Guidelines
Division, EPA v-
Dr. Walter Shackelford, ACB
Dr. Ron Webb, ACB
154
-------
Analysis of Consent Decree Pollutants
Various researchers and contractors involved in the analysis
of the base-neutral extractable priority pollutants have
noted that both the GC retention time and the mass spectral
fragmentation pattern of N-nitrosodiphenylamine (one of the
priority pollutants) and diphenylamine axe apparently
identical. Our studies on this problem have shown that as
N-nitrosodiphenylamine (mp 67 C) is heated it begins to
decompose as soon as it is in the liquid state. Above 145 C
the decomposition proceeds very quickly yielding diphenyla-
mine and tetraphenylhydrazine. The identities of these two
compounds were established by infrared and mass spectral
data.
When N-nitrosodiphenylamine is subjected to gas chromatography
under the conditions imposed by the Consent Decree Protocol,
i.e., inlet temperature 275 C, decomposition occurs in the
inlet, resulting in a single sharp symetrical peak that has
been identified as diphenylamine (elution temperature 161°C,
RRT 0.97). This compound is formed in 40 to 80% yield.
Formation of tetraphenylhydrazine in the GC inlet may also
occur, but has not been established since this compound does
not elute under the protocol conditions. No GC peak that
could be identified as N-nitrosodiphenylamine has to date
been observed.
Identification of this nitrosamine via the regimen of the
protocol is inconclusive and it is therefore suggested that
the apparent presence of this compound be currently reported
as "N-nitrosodiphenylamine and/or diphenylamine" until a
valid analytical method can be developed. We are investigating
liquid chromatography as a separation tool for nitrosamines,
including N-nitrosodiphenylamine.
Preliminary work with 1,2-diphenylhydrazine (hydrazobenzene—
another priority pollutant) indicates that it also degrades,
perhaps not in the GC inlet, but definitely in solution,
forming azobenzene as the major product, along with aniline
and an unknown of mw 184. Current data have eliminated
benzidine as the unknown's identity and indicate that it
might be a N-phenylphenylenediamine. This decomposition
occurs in methanol and methylene chloride (the solvent for
the protocol standards), as well as in water. Additional
work is needed on this problem before any concrete recommenda-
tions can be made. (Alfred F. Haeberer, 404-546-3187, FTS-
250-3187).
155
-------
Index of References
1. letter: Examples of trap packing deterioration. NUS,
Miss C. Ellen Gonter, November 15, 1977.
2. A Brief Evaluation of Phenol Extraction Procedures. Environ-
mental Science and Engineering, Inc., November 9, 1977.
3. Analytical Problems in Effluent Analysis, Dow Chemical Company,
R.O. Kagel.
4. Draft - Priority Pollutant Validation Protocol, Dow Chemical Co.
R.O. Kagel and R. H. Stehl.
5. Memo, dated November 23, 1977, Subject: Metals Analysis-
Chicago Regional Laboratory.
6. Preliminary Interim Procedures for Fibrous Asbestos, Charles H.
Anderson and J. MacArthur Long, U.S. E.P.A., Environmental
Research Laboratory, Athens, Georgia.
7. Analytical Methodology for the Determination of Asbestos by
Transmission Election Microscopy. Walter C. McCrone Assoc.,
Inc.
8. Preservation of Phenolic Compounds in Wastewaters, M.J. Carter
and M.T. Huston, E.P.A., Central Regional Laboratory.
9. Diagram of Liquid-Liauid extractor, MIDWEST Research Institute,
C.L. Haile.
156
-------
IMUS
CORPORATION
CYRUS WM. RICE DIVISION
November 15, 1977
ANALYTICAL SERVICES LABORATORY
15 NOBLE AVENUE • PITTSBURGH. PA. 15205
Mr. William A. Telliard
Chief, Electric Utilities &
Mining Branch
Effluent Guidelines Division
U.S. Environmental Protection Agency
Waterside Mall
401 M Street, S.W.
Washington, D.C. 20460
Dear Bill:
Enclosed are examples of what is believed to be trap packing deterioration.
1. February 2, 1977, 5.0 ml of sample was purged according to the Bellar-
Lichtenberg procedure (EPA-670/4-74-009). The trap was sealed with
stainless steel caps and frozen. July 14, 1977, the trap was allowed
to warm to room temperature and run on GC/MS. The trap was sealed
with Teflon caps, and stored in a drawer at ambient temperature. July
21, 1977, the trap was desorbed onto the GC column, and the resultant
curve 1 obtained.
2. July 21, 1977, 5.0 ml of the retain sample was purged, trapped, desorbed,
etc. according to the Bellar-Lichtenberg procedure, (March 1977 trap
packing) and the resultant curve 2 obtained.
This was not an isolated case.
The peaks at 5.3 and 13.5 have appeared in most of the traps that were frozen.
The peak at 15.5 "grows" larger with time in an eight-hour period, even
though the trap is heated at 180°C with nitrogen at 40 ml/min flowing through
it for 20 to 30 minutes between runs..
At present we are using one trap per day, following the procedure as written
(some samples are diluted before purging), and not using the freezing technique.
Sincerely,
(Miss) C. Ellen Gonter, Manager
Water Laboratories Department
Enclosure
157
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environ mental srivni'p and
P.O. BOX 13454 9 GAINESVILLE, FLORIDA 32604
szc.
904/372-3318
75-054-104
A BRIEF EVALUATION OF
PHENOL EXTRACTION PROCEDURES
Prepared by:
ENVIRONMENTAL SCIENCE AND ENGINEERING, INC.
P. 0. BOX 13454, UNIVERSITY STATION
GAINESVILLE, FLORIDA 32604
NOVEMBER 9, 1977
For:
EFFLUENT GUIDELINES DIVISION
ENVIRONMENTAL PROTECTION AGENCY
WASHINGTON, D.C. 20460
ATLANTA , GEORGIA
404 / 688-502S
JACKSONVILLE , FLORIDA
904 / 398-8303
ST. LOUIS , MISSOURI
314 / 567-4600
TAMPA , FLORIDA
313 / 886-6672
160
-------
Introduction
This report is the end result of a very brief study on the effectiveness
of the current protocol method for extraction and analysis of the acidic
(phenolic) fraction of the semi-volatile extractables. Alternative methods were
also investigated and the results are discussed here.
It should be noted that this study, which was designed and executed
in about 150 man hours, is in no way conclusive. The presentation of the
data in this report is for discussion purposes and to help in the solution
of a very complex problem.
161
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Procedures for Phenol Analysis
I Base/Neutral and Acidic Extraction (Similar to EPA Protocol)
1) The sample should be preserved with CuSO and phosphoric acid
to pH=4 as in Standard Methods, 14th Edition, p. 576.
2) Measure 100 ml of the sample into a 250 ml separatory funnel.
3) Adjust the pH of the sample to pH=12 with 6N NaOH solution.
4) Extract the sample with 50 ml of methylene chloride. Shake
for 2 min. and let emulsion break. Repeat the extraction with
25 ml and 25 ml of methylene chloride. Save the methylene
chloride layers for base-neutral analysis.
5) Adjust the pH of the sample to pH=2 with concentrated HC1 solution.
6) Extract with 50 ml, 25 ml, 25 ml of methylene chloride. Transfer
the methylene chloride layers through a 75 mm diameter glass funnel
containing a glass wool plug and a 2 cm layer of anhydrous sodium
sulfate to a 500 ml Kuderna-Danish apparatus with a 10 ml receiver.
7) Evaporate the extract down to 1.0 ml.
8) Add 10 ul of a 2 ug/ul d-^-anthracene internal standard solution to
the 1.0 ml of the extract.
9) Inject 2 ul of the extract on the gas chromatograph/mass spectrometer
using the GC conditions given in part II.
10) Calculate the amount of phenols in the sample using relative response
factors with respect to the dj_Q-anthracene internal standard obtained
from standard solutions.
162
-------
II Steam Distillation and Extraction Method
1) The sample should be preserved with CuSO and phosphoric acid
to pH=4 as in Standard Methods, 14th Edition, p. 576.
2) Measure 500 ml of the sample into the 1 liter boiling flask
of the distillation apparatus. Add several glass boiling beads.
3) Distill 450 ml of the sample, stop the distillation and when
boiling ceases add 50 ml phenol-free distilled water to the
distilling flask. Continue distillation until a total of 500 ml has
been collected.
4) Transfer the distillate to a 1 liter separatory funnel washing
the distillate container with several rinses of distilled water.
Combine the washings into the separatory funnel.
5) Adjust the pH of the distillate to pH=12 with 6N NaOH solution.
Check the pH with pH paper. Extract with 250 ml, 100 ml, 100 ml
of methylene chloride. Each time the separatory funnel should be
shaken for 2 minutes. Let the layers separate and draw off and
discard the methylene chloride layer.
6) Adjust the pH of the remaining aqueous distillate in the separatory
funnel to pH=2 with concentrated HC1. Check pH with pH paper.
7) Extract with 200 ml, 100 ml, 100 ml of methylene chloride. The
funnel should be shaken for at least 2 minutes each time.
8) Draw off the methylene chloride layers and pass through a glass
funnel containing a small amount of anhydrous sodium sulfate into
a 500 ml Kuderna-Danish apparatus with a 10 ml receiver.
9) Add a glass boiling bead and concentrate the combined methylene
chloride extracts to 1.0 ml on a boiling water bath.
10) Add 10 ul of a 20 ug/ul solution of d-, Q-anthracene to the 1.0 ml extract.
163
-------
II Continued
11) Inject 2 ul of the sample onto a gas chromatographic column using
the conditions given below.
MS/GC conditions:
Column: 6 ft x 2 mm i.d. glass
Packing: Tenax GC, 60/80 mesh
Flow: 30 tnl/min, Helium
Column Temperature: 180°C to 300°C at 8°C/min
Injector Temp.: 250°C
Jet Temp: 290°C
Transfer Line: 290°C
12) Calculate the amount of phenols in the sample using mass
spectrometer detection based upon relative response factors with
respect to d,Q-anthracene obtained from standard solutions.
164
-------
TABLE I
Distilled Water Spiking Experiments
Compound
Acidic Extraction
Only
92.5
89.1
1 88.2
88.4
.enol 89.6
57.7
sol 97.5
87.5
%RSD
11.4
5.6
5.1
0.8
1.5
0.6
3.7
1.7
II
1) Base Neutral Extraction
2) Acidic Extraction
III
1) Steam Distillation
2) Acidic Extraction
Phenol
0-Chlorophenol
0-Nitrophenol
2,4-Dichlorophenol
4-Chloro-m-cresol
2,4,6-Trichloroph
2,4-Dinitrophenol
p-Nitrophenol
4,6-Dinitro-o-cresol
Pentachlorophenol
I and II} 1 liter of water spiked with 100 ug of each phenol
III } 500 ml of water spiked with 100 ug of each phenol
Phenol and 2,4-Dinitrophenol were not included in standard solution
Creosote Waste Spiking Experiments
% Recovery
88.9
90.4
91.2
87.9
94.9
56.5
95.4
89.5
%RSD
8.0
6.1
7.0
4.4
5.7
5.1
8.4
5.7
75.7
73.5
78.2
85.9
78.1
3.1
67.8
80.7
%RSD
5.4
6.7
6.2
4.9
8.4
0
16.8
5.7
IV
1) Base-Neutral Extraction
2) Acidic Extraction
V
90.7
88
85
77
88
76
47
76
67
93
% Recovery
%RSD
9.2
17.
4.2
4.7
22.
11.7
16.5
10.7
3.
12.
1) Steam Distillation
2) Base-Neutral 3) Acidic Extract
Phenol
0-Chlorophenol
0-Nitrophenol
2,4-Dichlorophenol
4-Chloro-m-cresol
2,4,6-Trichlorophenol
2,4-Dinitrophenol
p-Nitrophenol
4,6-Dinitro-o-cresol
Pentachlorophenol
IV } 1 liter of water spiked with 1 ing of each phenol
V } 500 ml of water spiked with 1
-------
PROPOSED ALTERNATE METHODS
III Liquid-Liquid Extraction Using Labeled Internal Standard
1) The sample should be preserved with CuSO, and phosphoric acid
to pH=4 as in Standard Methods, 14th Edition, p. 576.
2) Measure 500 ml of the sample into a 1 liter separatory funnel.
3) Prepare a stock solution in acetone of the labeled internal
standard (e.g. phenol-dg) containing 100 ug/ml of the labeled compound,
4) Spike the sample with an aliquot of the labeled internal standard
solution. (e.g. 10 ml x 100 ug/ml=1000ug)
5) Adjust the pH of the sample to pH=12 with 6N NaOH solution.
Add 250 ml of methylene chloride to the sample and shake very
gently for about 5 minutes. A gentle rolling action is used to
prevent emulsion formation. Draw off the methylene chloride
layer and repeat the extraction with 100 ml and 100 ml of methylene
chloride. Discard the methylene chloride layers.
6) To the remaining aqueous sample in the separatory funnel, add
concentrated hydrochloric acid to bring the pH to 2.
7) Extract the sample with 200 ml of methylene chloride using a
gentle rolling motion as before. Repeat the extraction with 100
ml and 100 ml more of methylene chloride. Transfer the extracts
through a glass funnel containing a small amount of Na^SO,
(anhydrous) into a 500 ml Kuderna-Danish apparatus which includes
a 10 ml receiver.
8) Concent-rate the methylene chloride extracts down to 1.0 ml on
a boiling water bath.
166
-------
Ill Continued
9) Inject 2 ul of the concentrate onto a gas chromatograph/mass
spectrometer using the conditions given in Procedure II.
10) Calculate the extraction efficiency of the labeled internal
standard based upon response factors determined from
standard runs. Correct the response for the other phenols
assuming the same extraction efficiency as the internal
standard.
11) a) From Standard Solution,
= Areao
™
FIS
W
IS
= response factor for internal standard
Area = observed MS response for internal standard
JL o
W = amount of internal standard injected (ng).
J- O
\ = response factor for phenolic component A,B,,
,...;
W, . = amount of component A,B,... injected (ng)
(,A,D ,. . .)
b) From Sample Extract Injection,
Area
Extraction Efficiency (EEf ) = IS_
for internal standard R . WTr.
FIS IS
Area = observed MS Area for internal standard in sample
injection
167
-------
(b) Continued
W = Amount of internal standard expected in sample
injection (ng)
(ug/ml) ppm, , = Area(A,B,...) x WIS x 500 x
(A'B"-0 AreaIS vT
WHERE,
ppm. = concentration of phenolic component
U,B,...) ln sample in (Ug/mi)
Area,, ^ = MS area of phenolic component in sample
\" »•£*)•••/ . . «
injection
Area = MS area of internal standard in sample injection
(.LS)
W = amount of internal standard expected in sample
injection (ng)
VI = volume of injection (ul)
500 = dilution factor
168
-------
ANALYTICAL PROBLEMS IN EFFLUENT ANALYSIS
R. 0. KAGEL
ENVIRONMENTAL SERVICES
DOW CHEMICAL COMPANY
628 BUILDING
MIDLAND, MICHIGAN
169
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ANALYTICAL PROBLEMS IN EFFLUENT ANALYSIS
The most frustrating aspect of analyzing a complex effluent stream
for specific organic compounds is the almost total lack of validated
analytical method for those compounds, in that media, at low
concentrations. Specifically, these analyses usually involve the
effluent stream at the point where it interfaces with public waters.
Here, the concentration of any specific organic compound is most
certainly well below the part per million level. A few comparative
definitions of parts per million (ppra), parts per billion (ppb), and
parts per trillion (ppt) are given in Figure 1 in order to put the
magnitude of the analytical problem into proper perspective.
At these levels it is extremely difficult, very tedious and usually
costly - but not impossible - to obtain statistically meaningful
analytical data. A statistically meaningful result can be obtained
only by using validated analytical methods. Analytical procedures
as such are not validated. Validation involves the statistical
treatment of the data to determine the accuracy, precision, sensitivity
and reproducibility of an analytical procedure from laboratory to
laboratory or even from analyst to analyst within a laboratory. In
other words, validation provides a common denominator for agreement
on what an analytical result really means.
During the last five years, many industrial and government laboratories
have been busy developing analytical procedures for determining trace
levels of specific organic compounds in aqueous media. The increased
activity in this direction is the result of two things. First (Figure 2),
is the stagnation of analytical technology associated with those
methods that represent the shot gun approach to effluent analysis -
the BOD's, TOD's, TOC's etc. These methods provide gross parameters
that characterize the quality of the effluent and are used as control
parameters in most vasts treatment plants. The state of the art of
170
-------
this methodology has not changed appreciably over the past 10-15 years
ana for all practical purposes, this area of analytical technology has
become stagnant.
During the same time frame, significant state of the art developments
did occur in separations and detection technology. Gas chromatography-
inass spectrometry (GC-MS) is rapidly becoming a common place tool in
most analytical laboratories. Applied to effluent analyses, the
GC-MS represents the high powered rifle with a telescopic sight for
it allows the analytical chemist to zero-in on some specific compounds.
The combined use of separations technology, extraction of organic
components from a. waste water using an organic solvent such as hexane
or ether, followed by preconcentration and then detection by GC-MS
appears to be the universal approach to trace component analysis.
Figure 3 shows a typical example of this type of approach. Three
liters of a synthetic mixture of several compounds were extracted
with diethyl ether, preconcentrated by a factor of 3000 and analyzed
by GC-MS. All of these components are present at the ppb level.
Once the identity of each peak in the chromatogram has been established
by GC-MS, then subsequent analysis (Figure 4) are performed - in this
case - by election capture gas chromatography. This is simply to
avoid tying up a GC-MS which can range in price from $40K to $350K
with routine analysis which could easily be done on a $5K to $6K gas
chromatograph. The latter are readily available in most laboratories,
can easily be set up to do the analysis, and can be readily interfaced
with a computer to massage the data. These analytical procedures
for identifying and quantitating most of the components in an effluent
stream tend to be exceedingly tedious, time consuming and hence
quite costly. A good chrornotographer working in concert with a good
mass spectroscopist, given enough time,, the proper instrumentation,
a wide choice of column packings, will eventually develop an analytical
procedure for analyzing just about any system.
171
-------
A good example of Che kind of data generated by this approach is the
Environmental Protection Agency (EPA) study of the New Orleans area
water supply. Some 66 organic compounds, 10 of which are shown in
Figure 5, were reported to be present many at concentrations at or
less than 1 ppb. This study was one of the sources used, by EPA, to
develop the list of 65. The EPA research people who did this study
were very careful to emphasize that the values reported represent
highest concentration values rather than absolute values. This is
because when a component was determined by different methods, the
reported concentrations differed to some extent. Also, efficiency
values (recovery) for each stage of the analytical procedure were
not determined, i.e., the efficiency of carbon absorption of the
compound from water, losses incurred in drying the carbon, the efficency
of desorption, and losses incurred in concentrating the solvent to
low volumes. Without knowledge of these factors, one is hard pressed
to judge how good the results are because each step in the analytical
procedure introduces some error into the determination. The results
are probably good to ±50% at best and perhaps as much as ±100%, or
even more. The difference between 1 ppb and 2 ppb is probably not
significant in terms of the over all goals of the New Orleans study.
The New Orleans study was an exceptionally fine piece of work but
unfortunately it was not carried to completion - the procedures were
not validated. Hence it would be difficult for any two analysts to
produce numbers that would satisfactorily agree. It is obvious
that for the purpose of establishing effluent guidelines and monitoring
effluents one needs to establish better control and better technical
criteria on the analytical data. In general, analytical chemist
whether in industry or government are as genuinely concerned about
the accuracy, precision, sensitivity, and reproducibility of their
numbers as they are about developing the analytical procedures which
generate the numbers.
For 3. number of years, the residue analytical chemists were faced with
a similar analytical problem that involved the generation of meaningful
analytical data for pesticide residues in animal tissue, plants, soil
and water. Working together with USDA and FDA, they developed a mutually
172
-------
acceptable technical protocol for validating their analytical methods.
This protocol, the 10-10-10 principle, could easily be extended to
the case of effluent analysis, which in the broadest sense is a form
of residue analysis.
The mechanics of the 10-10-10 principle are shown in Figure 6. Ten
determinations are made on a control sample to determine interferences.
In residue studies the control is an untreated crop, soil, animal, etc.
A suitable control for effluent analysis is a synthetic sample of
plant effluent spiked with all compounds known to be present except
the one being analyzed. In this way the level of interference is
determined.
The fortified samples are used to determine recoveries. Samples of
control are usually spiked with the compound of interest at various
concentration and then spike is run through the entire analytical
procedure. This will show losses due to absorption on glassware,
charcoal absorption and desorption efficiencies, extraction efficiencies,
and the like. Generally, recoveries better than 85% are acceptable.
Realistically recoveries may range from 50% to 85%. The lower values
are acceptable if consistent results are obtained with replicate
samples.
Finally, 10 determinations are run on different aliquots of the same
samples to determine the precision of the procedure. The statistics
of the analytical procedure are usually verified by two independent
laboratories.
The 10-10-10 principle first surfaced in the 1950's. It was later
advocated by Harris and Cummings , USDA, (in 1964) as the absolute
minimum data requirement necessary to support the registration of a
pesticide use. In recent years the 10-10-10 principle has become
accepted protocol for validating analytical procedures in most
pesticide studies. It would be ironic if any data less than this
would be deemed adequate for drawing conclusions about effluent
studies.
173
-------
An example of the application of the 10-10-10 principle to a residual
herbicide metabolite in soil is shown in Figure 7. The chromatograms
represent a 5 ppb standard of the material, a soil control and the
control samples spiked at 5, 10, 100 and 500 ppb. The control is a
soil sample which has not been exposed to the herbicide.
Figure 8 shows the results of 10 determinations on 3 different
control soils. An interference is noted at the 0.4 ppb level. The
percent error at 2a (2 standard deviations) or the 95% confidence
level is ±50%. The blank is normally subtracted from recovery and
precision data. In this case, the blank is negligible.
The recovery data from spiked controls is shown in Figure 9. The
spike normally extends to 1/10 of the value of interest. The average
recovery shown here is 90% with an error of ±3% at the 2a level.
The small error is due to the large number of determinations, 25.
The error would have been larger if only 10 determinations had been
run.
As shown in Figure 10, the precision of the analytical procedure
based on 10 determinations of different aliquots of a sample each
containing about 500 ppb, is ±13%.
These statistics apply to any subsequent single operator determination
as long as there is no deviation from the method. For example, as
shown in Figure 11, a value of 484 ppb corrected for recovery and
blank translates into 537± 63 ppb.
This is of course an idealized example. Normally blanks are not
negligible, the recoveries are not 90% and the error may easily
range up to ±50%. However, once the procedure has been validated
any competent analytical chemist should be able to generate numbers
which are within the range of the stated errors.
174
-------
This type of validation scheme was recently used by Synons, et al ,
EPA Cincinnatti Labs, to determine sinsla operator precision and
accuracy for the determination of organohalides in chlorinated drinking
waters. The single operator precision for two replicate determinations,
shown in Figure 12, varies between 5 and 20% at the Ic? level. The
accuracy determined by two different laboratories on solutions of known
concentration, Figure 13, shows recoveries ranging, for example,
between 64 and 94% for a given compound. At the present, no known
data of this nature appears in the open literature for specific
organic compound in an effluent stream. There is a rather significant
difference between analyzing drinking water and an effluent stream.
The latter is a more complicated system and the procedure and method
that apply to drinking water are probably not transferable to effluent
analysis. The EPA is aware of this and is presently developing appropriate
analytical protocol for effluent analysis. Industry will be a contributing
party to the development of this protocol.
In conclusion, by applying validated analytical methods, statistically
meaningful values for the concentration of an organic compound in our
effluent can be obtained and, at least these numbers, mutually agreed
upon. Once the precision, accuracy, sensitivity, and reliability
of the methods has been established, it is then possible to establish
effluent guidelines, to monitor, B.A.T., and to affect protection of
the environment with a reasonable cost/benefit ratio.
References
1. Environmental News, Nov. 8, 1974; Draft Analytical Report; New
Orleans area water supply study. EPA, Lower Mississippi River
Facility, Region VI, S&A.
2. T. H. Harris & J. G. Cumniings, Residue Rev. _6, 104 (1964).
3. J. M. Symons et al, J. Am. Water Works Assoc. 67, 634 (1975).
175
-------
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FIGURE 2
ANALYSIS OF WASTE STREAMS
-STATE OF THE A R T -
GROSS PARAMETERS
BIOCHEMICAL OXYGEN DEMAND (BOD)
TOTAL OXYGEN DEMAND (TOD)
TOTAL ORGANIC CARBON (TOO
TOTAL DISSOLVED SOLIDS (TDS)
INDIVIDUAL COMPONENTS
SEPARATION TECHNOLOGY
LIQUID CHROMATOGRAPH (LC)
GAS CHROMATOGRAPH (GO
GAS CHROMATOGRAPH-MASS SPECTROMETRY (GC-MS)
177
-------
178
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FIGURE 4
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179
-------
FIGURE 5
COMPOUND
ORGANIC COMPOUND IDENTIFICATION
1
NEW ORLEANS AREA WATER SUPPLY STUDY
HIGHEST MEASURED CONCENTRATION yg/1 (ppb)
CARROLLTON JEFFERSON SI JEFFERSON =2
WATER PLANT WATER PLANT WATER PLANT
1 Acetaldehyde
D-VOA
NE
NE
Acetone
D-VOA
NE
3 AlkyIbenzene-C2 isoroer
4 Alkylbenzene-C2 i
5 AlkyIbenzene-C2 isomer
0.05
0.33
0.11
ND
ND
0.03
ND
ND
ND
6 ALkylbenzene-C3 isoner
0.01
ND
ND
7 AUcylbenzene-C3 isorner
0.04
0.05
0.02
isoner
0.02
ND
ND
Atrazine *
(2-chloro-4-ethylamino-
6-isopropylarnino-
s-triazine)
5.0
4.7
5.1
10 Deethylatrazine
(2- ch lore- 4-amino-
6-isopropylamino-
s-triazine)
0.51
0.27
0.27
180
-------
FIGURE 6
METHODS VALIDATION
THE "10-10-10" PRINCIPLE
10 DETERMINATIONS OF A CONTROL TO DETERMINE INTERFERENCES
10 DETERMINATIONS OF A FORTIFIED SAMPLE TO DETERMINE
RECOVERY VALUES
10 DETERMINATIONS OF ACTUAL SAMPLE TO DETERMINE PRECISION
OF THE METHOD
181
-------
FIGURE 7
J-
Q.
ro a.
o
^ §
-------
FIGURE 8
PRECISION OF BLANK
Concentration ppb
X.
1 0.5
2 0.3
3 0.4
4 0.2
5 0.3
6 0.4
7 0.4
8 0.3
9 0.4
10 0.5
X .4
E(Xj_ - X)2 = .1
a = ,/ KX, - X) 2 = .1
n - 1
Relative standard deviation at 951 confidence level
2 100% = 50%
+ 2£ 100%) = .4 + ,2ppb
X
183
-------
FIGURE 9
AGR
Number
113549
121954
122195
118092
127406
114598
121955
112239
117038
119526
114597
121954
122194
113424
132581
114596
113548
121921
130768
128288
130510
112240
122982
130767
128286
Location
Corvallis
Davis
Davis
Fargo
Fargo
Bozeman
Davis
Pendleton
Pendleton
Corvallis
Bozeman
Davis
Davis
Fargo
Bozeman
Bozeman
Corvallis
Davis
Bozeman
Pendleton
Fargo
Pendleton
Bozeman
Bozeman
Pendleton
ppb
Added Found
5 4.8
4.3
4.5
3.9
4.8
4.3
10 9.2
8.3
9.6
9.7
8.6
50 43.6
45.1
44.3
43.7
44.7
49.7
100 86.7
99.5
99.8
500 405
410
484
1000 888
973
Recovery
96
86
90
78
96
86
92
83
96
97
86
87
90
89
87
89
99
87
100
100
81
82
97
89
97
90 + 3*
*95% confidence limits for the mean.
AVERAGE PERCENTAGE RECOVERY
Rn = Rn = 90% = .90
n
Relative standard deviation at 95% confidence level of R
n
R
nfn"
R = 90+3%
n —
184
-------
FIGURE 10
PRECISION OF AN ANALYSIS
n Concentration ppb
Xi
1 436
2 451
3 410
4 484
5 447
6 451
7 443
8 ' 437
9 433
10 492
X = 448
Z(Xi - X)2 = 5210
a = V to ^xi "' = 24
n - 1
Relative standard deviation at 951 confidence level
2_£ 100% = 11%
X
X(1± X2g 100%) = 484 +53ppb
X
185
-------
FIGURE 11
ACTUAL CONCENTRATE
11%) = 537(1+ 11%) = 537 ± 63PPB
,90(1+ 3%)
CONC FOUND BLANK I RECOVERY CONCENTRATE AFTER
PPB PPB DETERMINED CORRECTION
484 ± 53 0,4+2, 90 + 3% 537 ± 63PPB
186
-------
FIGURE 12
DETERMINATION OF PRECISION3
Compound
LOW CONCENTRATION
Spiked Relative
Cone, yg/l a percent
HIGH CONCENTRATION
Spiked Relative
Cone, yg/l a percent
Chloroform
18
1,2-dichlor-
oethane
Carbon tet-
rachloride
14
Bromo-dichloro-
methane
20
Dibromo-chloro
methane
10
30
13
Bromoform
20
30
12
*Not determined at high concentration
187
-------
FIGURE 13
DETERMINATION OF ACCURACY5
(CONCENTRATION - yg/£)
Chloroform
1,2-Dichloro-ethane
Carbon Tetrachloride
Calculated
Lab A
Lab B
75 60_(+6-7%)
63(84) 46(77)
65(87) 46(77)
61(81) 54(90)
76(101)69(98)
10
5(+5%)
10
6(+14%)
9(90)
10(100)
10(100)
10(100)
6(120)
5(100)
5(100)
4(80)
9(90)
8(80)
S(80)
6(60)
5(83)
6(100)
6(100
4(67)
Calculated
Bromo-dichloro
methane
40
24(+5-7%)
Dibrcrro-chloro-rne thane
24
Bromoform
19(+10-13%) 40
23(+12-20%)
Lab A
Lab B
39(98) 22(92)
40(100)23(96)
35(88) 21(88)
38(95) 19(75)
23(96)
23(96)
17(71)
15(63)
14(74)
18(94)
13(68)
12(63)
40(100)
38(95)
48(120)
45(13)
18(78)
24(108)
24(104)
29(126)
188
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DRAFT
PRIORITY POLLUTANT VALIDATION PROTOCOL
R. 0. Kagel & H. H. Stehl
The Dow Chemical Company
Midland, Michigan 48640
1. ANALYTICAL METHODOLOGY
The methods for the priority pollutants are those listed
in "Analytical Methods for the Verification Phase of the Bat
Review" issued by Effluent Guidelines Division, Office of Water
and Hazardous Materials, U.S. Environmental Protection Agency.
Alternate Analytical methods will be considered if they are
prcperly substantiated in accordance with the following validation
Protocol. Methods must be described in sufficient detail in a
step-wise fashion that a competent analyst, unfamiliar with the specific
procedure can apply the method. Modifications of published methods
must be described fully. One method may suffice for simultaneous
analysis of several components.
2. VALIDATION PROTOCOL
The validation protocol is a modification of the EPA-EMSL
Analytical Quality Control Program1 and the Winter2 (EPA-EMSL)
interlaboratory validation study program. Each analytical
procedure must be validated by an adequate number of control
values and recovery values to establish the precision and
accuracy. Validation is necessary for an analytical procedure
to become an analytical method. The validation should be
repeated by at least three independent laboratories. Participating
laboratories must conform to the requirements specified by
Winter, in accordance with EPA-EMSL analytical quality control
programs, seven determinations for control and recovery values are
a minimal data requirement.
A. Best Achievable Limit of Detection (LOD)
The best achievable LOD'is obtained from the analyses of
seven samples of organic free water carried through the
entire analytical procedure. The observed peakrto-peak
noise, 0B, and the average peak-to-peak noise, 0B,
are determined. The best achievable limit of detection,
LOD-Q is defined as:
LODB = 2.5 6B
189
-------
Page 2
B. Control Values
Control values are th 2.5 DB, then LOD,
b. If 61
-------
Page 3
D. Precision Values
The precision of a single determination, 0s, at the 95% confidence
level is calculated from the recovery data as:
= 2 E (R? -
n - 1
E. Calculation of Data
The actual concentration, Ca, and the standard deviation,
tfr is: Ca + 231 = 0s (1 +_ 2 CTRs . 100%) - O1 (1 + 2 OlT . 1005
^-a ~ Ca -- ~
O
O1
5s (1 + 2 0Rs . 100%;
tt — ^f
Reporting of Data
a. Any result where 0s < LOD, is reported as "N.D. (LODj)
meaning, not detected, with the detection limit given
in parenthesis.
b. If LCD-,- ^ 0s < 4 LODj the result is reported as a
qualitative result. A second determination must be
run. The two separate results and an average will
be reported as quantitative results.
c. For 0s >, 4 LODj a single determination is reported
as a quantitative result.
REFERENCES
1. Handbook for Analytical Quality Control in Water and Wastewat
Laboratories EPA-EMSL 1976.
2. J. A. Winter, "Validation of Environmental Measurement
Methodology" Int. Conf. On Environmental Sensing and
Assessment, September 14-19, 1975.
191
-------
^v
INITED STATES ENVIRONMENTAL PROTECTION AGENCY
tfyi -
DATE:
SUBJECT: Metals Analysis - Chicago Regional Laboratory
William A. Tell lard, Chief
Energy and Mining Branch
Branch Chiefs, EGD
Project Officer, EGD
The following memo addresses a recent meeting that was held
between representatives of this office and several staff members
from the Region V Laboratory, with regards to the negotiation for
additional analytical support. Arrangements have been made for
an additional 1,000 samples to be analyzed by the Chicago
Regional Laboratory.
The following information pertains to the labeling, sampling and
type of containers to be utilized in the forth coming program for
metal analysis.
During the previous period of time, a number of points have been
raised regarding the use of uniformity in both the container and
sample size. The following notes should be made available to
contract personnel as well as the Surveillance and Analysis
Divisions:
1. Labeling Codes - Attachment A of this memo contains a
list of codes numbers to be used on labels which will be supplied
to you for those samples to be analyzed for metal parameters. A
code number shall be a six digit code, the first two digits
indicate the industrial category, the second two digits refer to
the contractor or sampling group and the third set is the sample
number. These codes and the labeling information are contained
in Appendix A. The regional lab will utilize this coding system
for their computer which handles the data output.
192
EPA FORM 1320-S (REV. 3-76) iz"~
-------
Example:
Be ~Mg Sn
TI
jCa. Hy V
11 22 57
Sampl e UoY
This code number means that this particular sample contains coal mining
water (11) taken by Versar (22) and that this is the 57th bottle
or sample taken.
2. Samples shall not be preserved with acid as it is
written in the Screening Protocol. This procedure in the only
way to comply with the Department of Transportation regulations
against shipping corrosive materials. Samples shall be prepared
for analysis of total metals by a hard digestion at the Chicago
Regional Labs. This means that a combination of nitric acid and
hydrochloric acid shall be added for samples analyzed by the
plasma unit.
3. Data Turnaround Time - Twenty-two elements can presently
be determined with the plasma unit. A number of parameters must
still be done by either flame!ess AA or flame AA for the purpose
of identification. To enable a better utilization of time, it is
recommended that the primary contractor (most of which have
atomic absorption capabilities) run the following parameters;
selenium, arsenic, antimony thallium and silver. Twenty-two
additional parameters can be supplied by the Central Regional Lab
with the plasma unit. This will cut down on the time delays due
to the limited instrumentation available in the laboratory.
4. Sample Type - As has previous been the case, samples
from the screening portion of the program shall be taken from the
composite sample (either influent or effluent or both) well mixed
and then put into a properly labeled container.
Additional sample capabilities will hopefully be made available,
some time after the first of the year. Until then, we will be
193
-------
limited to the 1,000 samples that have been negotiated. The need
for metals analysis for screening samples by all project officers
should be made known to myself or Gail Goldberg, as soon as
possible, so that scheduling can be afforded.
5. Quality Control - The Central Regional Laboratory will
continue to maintain a quality control file for all samples run
for EGD. This quality control file will be periodically supplied
to the Division and as needed can be incorporated into any court
record. The quality control file is probably the most complete
effort that the Division has been able to obtain. It specifies
the recoveries, performance of the instrument, and the individual
sample variability on a day-by-day basis. This information will
be made available through the Energy and Mining Branch to the
project officers and their contractors, as the need arises. The
quality control program at Central Regional Lab is far superior
to any program that has previously existed in the Division. It
can insure you that the metals analysis data are properly framed
and within the confines in definition of the performance
standards specified under 304(g). As each project comes to a
conclusion this data will be made available to the individual
project officers and Branches for inclusion in their record. A
period of a two weeks notice will be greatly appreciated, this
notification again, should be made in writing to Gail S. Goldberg
so that we may solicit the computer output for the quality
control data for those samples.
6. Samples, are collected as before meaning that there
exists only one sample for metal analysis per sampling site. The
possibility of collecting duplicates was considered. This idea
had to be rejected in view of time limits and financial
constraints.
7. The use of old labels for metals analysis, like the one
shown in the screening protocol should be discontinued. New
labels as shown in point 1. of this memo shall be distributed to
you. Only these new labels are compatible with the Chicago Lab
computer.
194
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Sampling Contractor Code Number
01. EPA Region I
02. EPA Region II
03. EPA Region III
04. EPA Region IV
05. EPA Region V
06. EPA Region VI
07. EPA Region VII
08. EPA Region VIII
09. EPA Region IX
10. EPA Region X
11. National Enforcement Investigations Center
12.
13. Hamilton Standard
14. Colin A. Houston & Associates
15. Environmental Science & Engineering , Inc.
16. Ryckman, Edgerley, Tomlinson and Associates, Inc.
17. E.H. Richardson Associates
18. Mid-West Research Instutute
19. NUS - Cyrus Rice Division
20. Burns & Roe, Inc.
21. Calspan Corporation
22. Versar Incorporated
23. Jacobs Engineering Company
24. E.G. Jordan Co., Inc.
25. Sverdrup 4 Parcel and Associates, Inc.
195
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26. Carborundum Corporation
27. TRW
28. Industrial Environmental Research Lab, Cincinnati
196
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Industrial Code Numbers
01 - Timber Products
02 - Steam Electric
03 - Leather Tanning
04 - Iron & Steel mfg.
05 - Petroleum Refining
06 - Nonferrous Metals
07 - Paving & Roofing
08 - Paint & Ink
09 - Printing & Publishing
10 - Ore Mining
11 - Coal Mining
12 - Organic Chemicals
13 - Inorganic Chemicals
14 - Textile Mills
15 - Plastics & Synthetics
16 - Pulp & Paper
17 - Rubber Processing
18 - Soaps & Detergents
19 - Auto & other Laundries
20 - Pesticides mfg.
21 - Photographic Industries
22 - Gum & Wood Industries
23 - Pharmaceuticals
24 - Explosives
25 - Adhesive & Sealants
26 - Battery mfg.
27 - Plastics mfg.
28 - Foundries
29 - Coil Coating
30 - Porcelain/Enameling
31 - Aluminum
32 - Copper
33 - Electronics
34 - Shipbuilding
35 - Electroplating
36 - Oil and Gas Extraction
197
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UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
SUBJECT: Sample Codes
FROM: William Telliard, Chief
Energy and Mining Branch
TO.- All EGO Project Officers
You will inform your sampling contractor to use the same digit coding
system for metals and organic samples. This six digit code system is
explained in the attached memo. Keeping the same code number for all
portions of samples greatly reduces the amount of data processing and
any changes for error in correlating samples.
198
EPA FORM 1320-6 (REV. 3-76)
-------
UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
DATE= NOV231977
SUBJECT: Chicago Lab - New address
FROM: William Telliard, Chief
Energy and Mining Branch
TO: All EGD Project Officers
The Chicago Lab has moved. The new address is:
U.S. Environmental Protection Agency
Region V, Central Regional Laboratory
536 South Clark
Chicago, Illinois 60605
Please send screening samples for metals analysis only, to the above
address.
EPA FORM 1320-6 (REV. 3-76)
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PRELIMINARY INTERIM PROCEDURE
FOR
FIBROUS ASBESTOS
by
Charles H. Anderson and J. MacArthur Long
Analytical Chemistry Branch
U.S. Environmental Protection Agency
Environmental Research Laboratory
College Station Road
Athens, Georgia 30601
-------
FIBROUS ASBESTOS
(Preliminary Interim Procedure)
(Transmission Electron Microscopy Method)
1. Scope and Application
1.1 This method is applicable to drinking water and water
supplies.
1.2 The method determines the number of asbestos
fibers/liter, their size (length and width), the size
distribution, and total mass. The method
distinguishes chrysotile from amphibole asbestos. The
detection limits are variable and depend upon the
amount of total extraneous particulate matter in the
sample as well as the contamination level in the
laboratory environment. Under favorable circumstances
0.1 MFL (million fibers per liter) can be detected.
The detection limit for total mass of asbestos fibers
is also variable and depends upon the fiber size and
size distribution in addition to the factors affecting
the total fiber count. The detection limit under
favorable conditions is in the order of 0.1 ng/1.
1.3 The method is not intended to furnish a complete
characterization of all the fibers in water.
1.4 It is beyond the scope of this method to furnish
detailed instruction in electron microscopy, electron
diffraction or crystallography. It is assumed that
those using this method will be sufficiently
knowledgeable in these fields to understand the
methodology involved.
1.5 The method outlined below is based upon what is
considered to be state-of-the art practice but it is
emphasized that at present no single analytical
procedure for asbestos is universally accepted. As a
result no inter-laboratory comparisons are presented
and the procedure should not be considered as a
standard method.
2. Summary of Method
2.1 A variable, known volume of water sample is filtered
through a membrane filter of sufficiently small pore
size to trap asbestos fibers. A small portion of the
filter with deposited fibers is placed on an electron
microscope grid and the filter material removed by
gentle solution in organic solvent. The material
remaining on the electron microscope grid is examined
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in a transmission microscope at high magnification.
The asbestos fibers are identified by their morphology
and electron diffraction pattern and their length and
width are measured. The total area examined in the
electron microscope is determined and the number of
asbestos fibers in this area is counted. The
concentration in MFL (millions of fibers/liter) is
calculated from the number of fibers counted, the
amount of water filtered, and the ratio of the total
filtered area/sampled filter area. The mass/liter is
calculated from the assumed density and the volume of
the fibers.
3. Definitions
Asbestos - A generic term applied to a variety of
commercially useful silicate minerals that may have a
fibrous structure.
Fiber - Any particle that has parallel sides and a
length/width ratio greater than or equal to 3:1.
Aspect Ratio - The ratio of length to width.
Chrysotile - A nearly pure hydrated magnesium silicate, the
fibrous form of the mineral serpentine, possessing a
unique layered structure in which the layers are
wrapped in a helical cylindrical manner about the
fiber axis.
Amphibole - A silicate mineral whose basic structural unit
is a double silica chain (Si^Ojj), but with a variable
composition and a layered structure that is easily
cleaved to form a fiber.
Detection Limit - The calculated concentration in MFL,
equivalent to one fiber above the background or blank
count.
Statistically Significant - Any concentration based upon a
total fiber count of five or more in 20 grid squares.
H. Sample Handling and Preservation
4.A Sampling
It is beyond the scope of this procedure to furnish detailed
instructions for field sampling; the general principles of
sampling waters are applicable. There are some
considerations that apply to asbestos fibers, a special type
of particulate matter. These fibers are small, and in water
range in length from .1 ym to 20 ym or more. Because of the
range of size there may be a vertical distribution of
particle sizes. This distribution will vary with depth
2
203
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depending upon the vertical distribution of temperature as
well as the local meteorological conditions. Sampling
should take place according to the objective of the
analysis. If a representative sample of a water supply is
required a carefully designed set of samples should be taken
representing the vertical as well as the horizontal
distribution and these samples composited for analysis.
U.1 Containment Vessel
The sampling container shall be a clean polyethylene,
screw-capped bottle capable of holding at least one
liter. The bottle should be rinsed at least two times
with the water that is being sampled prior to
sampling.
NOTE: Glass vessels are not suitable as sampling
containers.
ft. 2 Quantity of Sample
A minimum of approximately one liter of water is
required and the sampling container should not be
filled. It is desirable to obtain two samples from
one location.
4.3 Sample Preservation
No preservatives should be added during sampling and
the addition of acids should be particularly avoided.
If the sample cannot be filtered in the laboratory
within US hours of its arrival, sufficient amounts (1
ml/1 of sample) of a 2.71X solution of mercuric
chloride to give a final concentration of 20 ppm of Hg
may be added to prevent bacterial growth.
5. Interferences
5.1 Misidentification
The guidelines set forth in this method for counting
fibrous asbestos require a positive identification by
both morphology and crystal structure as shown by an
electron diffraction pattern. Chrysotile asbestos has
a unique tubular structure, usually showing the
presence of a central canal, and exhibits a unique
characteristic electron diffraction pattern. Although
halloysite fibers may show a similar streaking to
chrysotile they do not exhibit its characteristic
triple set of double spots or 5.3A layer line. It is
highly improbable that a non-asbestiform fiber would
exhibit the distinguishing chrysotile features.
Although amphibole fibers exhibit characteristic
morphology and electron diffraction patterns, they do
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not have the unique properties exhibited by
chrysotile. It is therefore possible though not
probable for misidentification to take place.
Hornblende is an amphibole and, in a fibrous form,
will be mistakenly identified as amphibole asbestos.
It is important to recognize that a significant
variable fraction of both chrysotile and amphibole
asbestos fibers do not exhibit the required
confirmatory electron diffraction pattern. This
absence of diffraction is attributable to unfavorable
fiber orientation and fiber sizes. The results
reported will therefore be low as compared to the
absolute number of asbestos fibers that are present.
5.2 Obscuration
If there are large amounts of organic or amorphous
inorganic materials present, some small asbestos
fibers may not be observed because of physical
overlapping or complete obscuration. This will result
in low values for the reported asbestos content.
5.3 Contamination
Although contamination is not strictly considered an
interference, it is an important source of erroneous
results, particularly for chrysotile. The possibility
of contamination should therefore always be a
consideration.
5.4 Freezing
The effect of freezing on asbestos fibers is not known
but there is reason to suspect that fiber break down
could occur and result in a higher fiber content than
was present in the original sample. Therefore the
sample should be transported to the laboratory under
conditions that would avoid freezing.
6. Equipment and Apparatus
6.1 Specimen Preparation Laboratory
The ubiquitous nature of asbestos, especially
chrysotile, demands that all sample preparation steps
be carried out to prevent the contamination of the
sample by air-borne or other source of asbestos. The
prime requirement of the sample preparation laboratory
is that it be sufficiently free from asbestos
contamination that a specimen blank determination
using 200 ml of asbestos-free water yields no more
than 2 fibers in twenty grid squares of a conventional
200 mesh electron microscope grid.
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In order to achieve this low level of contamination,
the sample preparation area should be a separate
conventional clean room facility. The room should be
operated under positive pressure and have incorporated
electrostatic precipitators in the air supply to the
room, or alternatively absolute (HEPA) filters. There
should be no asbestos floor or ceiling tiles, transite
heat-resistant boards/ nor asbestos insulation. Work
surfaces should be stainless steel or Formica or
equivalent. A laminar flow hood should be provided
for sample manipulation. Disposable plastic lab coats
and disposable overshoes are recommended.
Alternatively new shoes for all operators should be
provided and retained for clean room use only. A mat
(Tacky Mat, Liberty Industries, 589 Deming Rd.,
Berlin, Connecticut 06037, or equivalent) should be
placed inside the entrance to the room to trap any
gross contamination inadvertently brought into the
room from contaminated shoes. Normal electrical and
water services, including a distilled water supply
should be provided. In addition a source of ultra-
pure water from a still or filtration-ion exchange
system is desirable.
6.2 Instrumentation
6.2.1 Transmission Electron Microscope. A
transmission electron microscope that operates
at a minimum of 80 KV, has a resolution of 1.0
nm and a magnification range of 300 to
100,000. If the upper limit is not attainable
directly it may be attained through the use of
auxiliary optical viewing. It is mandatory
that the instrument be capable of carrying out
selected area electron diffraction (SAED) on
an area of 300 nm. The viewing screen shall
have either a millimeter scale, concentric
circles of known radii, or other devices to
measure the length and width of the fiber.
Most modern transmission microscopes meet the
requirements for magnification and resolution.
An energy-dispersive X-ray spectrometer is
useful for the identification of suspected
asbestiform minerals; this accessory to the
microscope, however, is not mandatory.
6.2.2 Data Processor. The large number of
repetitive calculations make it convenient to
use computer facilities together with
relatively simple computer programs.
206 5
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6.2.3 Vacuum Evaporator. For depositing a layer of
carbon on the Nuclepore filter, and for
preparing carbon coated grids.
6.2.4 Low Temperature Plasma Asher. To be used for
the removal of organic material (including the
filter) from samples containing so much
organic matter that asbestos fibers are
obscured. The sample chamber should be at
least 10-cm diameter.
6.3 Apparatus* Supplies and Reagents
6.3.1 Jaffe Wick Washer. For dissolving Nuclepore
filter (if Nuclepore is used in sample
preparation). Assemble as in 8.2A.1. It is
illustrated in Figure 1.
6.3.2 Condensation Washer. For use in dissolving
the Millipore filter when using the Millipore
sample preparation method. A system with
controlled heating, controlled refluxing, and
a cold finger for holding the electron
microscope (EM) sample grids. At least two
systems are commercially available. Figure 2
is an illustration of one design that has
proven satisfactory.
6.3.3 Filtering Apparatus. 47-mm funnel (Cat. No.
XX1504700, Millipore Corporation, Order
Service Dept., Bedford, MA 01730). Used to
filter water samples. 25-mm funnel (Millipore
Cat. No. XX1002500). Used to filter dispersed
ash samples.
6.3.1 Vacuum Pump. For use in sample filtration.
Should provide vacuum up to 20 inches of
mercury.
6.3.5 EM Grids. 200-mesh copper or nickel grids,
covered with carbon-coated collodion for use
with the Millipore-condensation washing
technique. Formvar-backed grids, without a
carbon coating are used in the Nuclepore-Jaffe
sample preparation method. These grids may be
purchased from manufacturers of electron
microscopic supplies or prepared by standard
electron microscopic grid preparation
procedures. Finder grids may be substituted
and are useful if the re-examination of a
specific grid opening is desired.
207
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Screen Support
with Grid
Ridge
Petri Dish
Glass Slides
A.
Layer of Filter Papers
Nuclepore Filter
Carbon
Chloroform
Formvar
Grid
Carbon
Grid
B.
Figure 1. Modified Jaffe Wick Method
A. Washing Apparatus
B. Washing Process
208
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1.70 mm
\
3.30 mm
3.14 mm
Condenser
RJ
25 min
View A
Water Source
X
Brass Holder
(See View A)
Cold Finger
Adapter
Heating Mantle
Powerstat
_2 mm
Water Drain
•Figure 2. Condensation Washer
209
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6.3.6 Membrane Filters.
47-mm diameter Millipore membrane filter, type
HA; 0.45 urn pore size. For filtration of
water sample.
47-mm diameter Nuclepore membrane filter; 0.1
ym pore size. (Nuclepore Corp, 7035 Commerce
Circle, Pleasanton, CA 94566) For filtration
of water sample.
47-mm diameter Millipore membrane filter type
BS; 2 ym pore size. Used as a Nuclepore filter
support on top of the glass frit.
25-mm diameter Millipore membrane filter, type
HA; 0.45 ym pore size. To filter dispersed
ashed Millipore filter.
25-mm diameter Nuclepore membrane filter; 0.1
ym pore size. To filter dispersed ashed
Millipore filter.
25-mm diameter Millipore membrane filter, type
BS; 2.0 ym pore size. To be used as a
Nuclepore filter support on top of the glass
frit.
6.3.7 Glass Vials. 30-mm diameter x 80-rran long.
For holding filter during ashing.
6.3.8 Glass Slides. 5.1-cm x 7.5-cm. For support
of Nuclepore filter during carbon evaporation.
6.3.9 Scalpels. With disposable blades and
scissors.
6.3.10 Tweezers. Several pairs for the many handling
operations.
6.3.11 "Scotch" Doublestick tape. To hold filter
section flat on glass slide while carbon
coating.
6.3.12 Disposable Petri dishes, 50-mm diameter, for
storing membrane filters.
6.3.13 Static Eliminator, 500 microcuries Po-210.
(Nuclepore Cat. No. V090POL00101) or
equivalent. To eliminate static charges from
membrane filters.
6.3.14 Carbon rods, spectrochemically pure, 1/8" dia.,
3.6 mm x 1.0 mm neck. For carbon coating.
9
210
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6.3.15 Carbon rod sharpener. (Cat. No. 1204, Ernest
F. Fullam, Inc., P. O. Box 444, Schenectady,
NY 12301) For sharpening carbon rods to a
neck of specified length and diameter.
6.3.16 Ultrasonic Bath. (50 watts, 55 KHz). For
dispersing ashed sample and for general
cleaning.
6.3.17 Graduated Cylinder, 500 ml.
6.3.18 Spot plate.
6.3.19 10 yl Microsyringe. For administering drop of
solvent to filter section during sample
preparation.
6.3.20 Carbon grating replica, 2160 lines/mm. For
calibration of EM magnification.
6.3.21 Cork borer (1/8 inch diameter). For sampling
prepared Millipore filters.
6.3.22 Filter paper. S & S #589 Black Ribbon or
equivalent (9-cm circles). For preparing
Jaffe Wick Washer.
6.3.23 Screen supports (copper or stainless steel) 12
mm x 12 mm, 200 mesh. To support specimen
grid in Jaffe wick Washer.
6.3.24 Brass holder. For holding specimen in
condensation washer. See Figure 2, View A.
6.3.25 Chloroform, spectro grade, doubly distilled.
For dissolving Nuclepore filters.
6.3.26 Acetone, reagent grade or better. For
dissolving Millipore filters,
6.3.27 Asbestos. Chrysotile (Canadian), Crocidolite,
Amosite. DICC (Onion Internationale Centre le
Cancer) Standards. Available from Duke
Standards Company, 445 Sherman Avenue, Palo
Alto, CA 94306.
6.3.28 Petri dish, glass (100 mm diameter x 15 mm
high). For modified Jaffe Wick Washer.
6.3.29 Alconox. (Alconox, Inc., New York, NY 10003)
For cleaning glassware. Add 7.5 g Alconox to
a liter of distilled water.
10
211
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6.3.30 Aerosol OT, 0.1X solution (Cat. No. So-A-292,
Fisher Scientific Company, 711 Forbes Avenue,
Pittsburgh, PA 15219) Used as dispersion
medium for ashed Millipore filter. Prepare a
0.1% solution by diluting 1 ml of the 10$
solution to 100 ml with distilled water.
Filter through 0.1-ym Nuclepore filter paper
before using.
6.3.31 Parafilm. (American Can Company, Neenah, WI)
Used as protective covering for clean
glassware.
6.3.32 Pipets, disposable, 5 ml and 50 ml.
6.3.33 Distilled or deionized water. Filter through
0.1-ym Nuclepore filter for making up all
reagents and for final rinsing of glassware,
and for preparing blanks.
6.3.3* Mercuric chloride, 2.71* solution w/v. Used
as sample preservative. See 4.3. Add 5.42 g
of reagent grade mercuric chloride (HgCl2) to
100 ml distilled water and dissolve by
shaking. Dilute to 200 ml with additional
water. Filter through 0.1-pm Nuclepore filter
paper before using.
7. Preparation of Standards
Reference standard samples of asbestos that can be used for
quality control for a quantitative analytical method are not
available. It is, however, necessary for each laboratory to
prepare at least two suspensions; one of chrysotile and
another of a representative amphibole. These suspensions
can then be used for intra-laboratory control and furnish
standard morphology photographs and diffraction patterns.
7.1 Chrysotile Stock Solution.
Grind about 0.1 g of UICC chrysotile in an agate
mortar for several minutes, or until it appears to be
a powder. Weigh out 10 mg and transfer to a clean 1
liter volumetric flask, add several hundred ml of
millipore filtered distilled water containing 0.1
percent Aerosol OT and one ml of a 20,000 ppm solution
of mercury and then make up to 1 liter with the 0.1
percent Aerosol filtered distilled water. To prepare
a working solution, transfer 10 ml of the above
suspension to another 1-liter flask, add 1 ml of a
20,000 ppm solution of mercury and make up to 1 liter
with the same 0.1 percent aerosol OT solution. This
suspension contains 100 yg per liter. Finally
transfer 1 ml of this suspension to a 1-liter flask,
11
212
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add 1 ml of a 20,000 ppm solution of mercury and make
up to volume with the 0.1 percent aerosol OT solution.
The final suspension will contain 5-10 MFL and is
suitable for laboratory testing.
7.2 Amphibole Stock Dispersion.
Prepare amphibole suspensions from UICC amphibole
samples as in Section 7.1.
7.3 Identification Standards
Prepare electron microscopic grids containing the UICC
asbestos fibers according to 8, Procedure, and obtain
representative photographs of each fiber type and its
diffraction pattern for future reference.
8. Procedure
8.1 Filtration.
The separation of the insoluble material, including
asbestiform minerals, through filtration and
subsequent deposition on a membrane filter is a very
critical step in the procedure. The objective of the
filtration is not only to separate, but also to
distribute uniformly the particulate matter such that
discreet particles are deposited with a minimum of
overlap.
The volume filtered will range from 50-500 ml. In an
unknown sample the volume can not be specified in
advance because of the presence of variable amounts of
particulate matter. In general sufficient sample is
filtered such that a very faint stain can be observed
on the filter medium. The maximum loading that can be
tolerated is 20 yg/cm2, or about 200 yg on a 47-mm
diameter filter; 5 yg/cm2 is near optimum. If the
total solids content is known, an estimate of the
maximum volume tolerable can be obtained. In a sample
of high solids content, where less than 50 ml is
required, the sample should be diluted with filtered
distilled water so that a minimum total of 50 ml of
water is filtered. This step is necessary to allow
the insoluble material to deposit uniformly on the
filter. The filtration funnel assembly must be
scrupulously clean and cleaned before each filtration.
The filtration should be carried out in a laminar flow
hood.
NOTE 1: The following cleaning procedure has been
found to be satisfactory:
12
213
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Wash each piece of glassware three times with
distilled water. Following manufacturer's
recommendations use the ultrasonic bath with an
Alconox-water solution to clean all glassware. After
the ultrasonic cleaning rinse each piece of glassware
three times with distilled water. Then rinse each
piece three times with deionized water which has been
filtered through 0.1-ym Nuclepore filter. Dry in an
asbestos-free oven. After the glassware is dry, seal
openings with parafilm.
8.1.1 Filtration
a. Assemble the vacuum filtration apparatus
incorporating either the . 1-um Nuclepore
backed with 2-^m Millipore, or the . 45-ym
Millipore filter. See 8.2A. 2 or 8.2B.2.
b. Vigorously agitate the water sample in its
container.
c. If the required filtration volume can be
estimated, either from turbidity estimates of
suspended solids or previous experience,
immediately withdraw the proper volume from
the container and add the entire volume to the
47-mm diameter funnel. Apply vacuum
sufficient for filtration but gentle enough to
avoid the formation of a vortex. If a
completely unknown sample is being analyzed, a
slightly modified procedure must be followed.
Pour 500 ml of a well-mixed sample into a 500
ml graduated cylinder and immediately transfer
the entire contents to the prepared vacuum
filtration apparatus. Apply vacuum gently and
continue suction until all of the water has
passed through the filter. If the resulting
filter appears obviously coated or discolored,
it is recommended that another filter be
prepared in the same manner, but this time
using only 200 or 100 ml of sample.
NOTE 1: Do not add more water after
filtration has started and do not rinse the
sides of the funnel.
d. Disassemble the funnel, remove the filter
and dry in a covered petri dish.
8.2 Preparation of Electron Microscope Grids.
The preparation of the grid for examination in the
microscope is a critical step in the analytical
procedure. The objective is to remove the organic
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filter material from the asbestos fibers with a
minimum loss and movement and with a minimum breakage
of the grid support film. Two alternative procedures
are acceptable:
A. Nuclepore Filter, Modified Jaffe Wick
B. Millipore Filter, Condensation Washer
If the sample contains organic matter in such amounts
that interfere with fiber counting and identification
a preliminary ashing step is required. See 8.5.
NOTE 1: Two alternatives for grid preparation are
suggested because the superiority of one technique
over the other has not been substantiated by
sufficient experimental evidence. The differences
between the two techniques of sample preparation lie
in the filtering medium (Nuclepore vs. Millipore) ,
whether the filter is carbon coated, and in the method
of dissolving the filter material. There is evidence
that the condensation washing procedure can lose
amphibole fibers and that amphiboles are more
susceptible to loss than chrysotile.
8.2A Nuclepore Filter, Modified Jaffe Wick Technique.
8.2A.1 Preparation of Modified Jaffe Washer
Place three glass microscope slides (75 mm x
25 mm) one on top of the other in a petri dish
(100 mm x 15 mm) along a diameter. Place 14 S
& S #589 Black Ribbon filter papers (9-cm
circles) in the petri dish over the stack of
microscope slides. Place three mesh copper
screen supports (12 mm x 12 mm) along the
ridge formed by the stack of slides underneath
the layer of filter papers. Place an EM
specimen grid on each of the screen supports.
See Fig. 1.
8.2A.2 Vacuum Filtration Unit
Assemble the vacuum filtration unit. Place a
2-ym Millipore filter type BS on the glass
frit and then position a 0.1-ym Nuclepore
filter, shiny side up, on top of the Millipore
filter. Apply suction to center the filters
flat on the frit. Attach the filter funnel
and shut off the suction.
8.2A.3 Sample Filtration
See 8.1.1.
14
215
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8.2A.U Sample Drying
Remove the filter funnel and place the
Nuclepore filter in a loosely covered petri
dish to dry. The petri dish containing the
filter may be placed in an asbestos-free oven
at 45° C for 30 minutes to shorten the drying
time.
8.2A.5 Selection of section for carbon coating
Using a small pair of scissors or sharp
scalpel cut out a retangular section of the
Nuclepore filter. The minimum approximate
dimensions should be 15 mm long and 3 mm wide.
Avoid selection near the perimeter of the
filtration area.
8.2A.6 Carbon Coating the Filter
Tape the two ends of the selected filter
section to a glass slide using "Scotch" tape.
Take care not to stretch the filter section.
Identify the filter section using a china
marker on the slide. Place the glass slide
with the filter section into the vacuum
evaporator. Insert the necked carbon rod and,
following manufacturer's instructions, obtain
high vacuum. Evaporate the neck, with the
filter section rotating, at a distance of
approximately 7.5 cm from the filter section
to obtain a 30-50 nm layer of carbon on the
filter paper. Evaporate the carbon in several
short bursts rather than continuously to
prevent overheating the surface of the
Nuclepore filter.
NOTE 1: Overheating the surface tends to
crosslink the plastic, rendering the filter
dissolution in chloroform difficult.
NOTE 2: The thickness of the carbon film can
be monitored by placing a drop of oil on a
porcelain chip that is placed at the same
distance from the carbon electrodes as the
specimen. Carbon is not visible in the region
of the oil drop thereby enabling a visual
estimate of the deposit thickness by the
contrast differential.
8.2A.7 Grid Transfer
Remove the filter from the vacuum evaporator
and cut out three sections somewhat less than
15
216
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3 mm x 3 mm and such that the square of
Nuclepore fits within the circumference of the
grid. Pass each of the filter sections over a
static eliminator and then place each of the
three sections carbon-side down on separate
specimen grids previously placed in the
modified Jaffe Washer. Using a microsyringe,
place a 10-yl drop of chloroform on each
filter section resting on a grid and then
saturate the filter pad until pooling of the
solvent occurs below the ridge formed by the
glass slides inserted under the layer of
filter papers. Place the cover on the petri
dish and allow the grids to remain in the
washer for approximately 24 hours. Do not
allow the chloroform to completely evaporate
before the grids are removed. To remove the
grids from the washer lift the screen support
with the grid resting upon it and set this in
a spot plate depression to allow evaporation
of any solvent adhering to the grid. The grid
is now ready for analysis or storage.
8.2B Millipore - Condensation Washer Technique
8.2B.1 Operation of Condensation Washer
Fill the extractor flask to UQ% capacity with
acetone. Filter the acetone through 0.1-ym
Nuclepore filter paper before using. Adjust
the tap water flow rate to 10 ml/sec by
allowing the water exiting from the cold
finger to run into a graduated cylinder for 30
seconds. Set the variable transformer,
regulating the heater power-input, to
approximately 45 volts. Sufficient heat
should be applied to generate acetone vapors
at the required condensation or reflux level
without boiling or simmering. The reflux
level should be even with the top of the cold
finger and just below the stainless steel
grid. (Important: See Note 1). After the
system has been running for twenty minutes,
check the reflux level of the acetone. Place
a heavily lined index card behind the adaptor
so that when viewed from the front of the
adaptor the cold finger is parallel to the
heavy lines on this card. Locate the reflux
level by noting the illusionary wavy motion of
the heavy lines on the index card. If the
reflux level is too high, increase, or if too
low, decrease the tap water flow rate. Do not
allow pooling of the solvent to occur on the
grids. At very low flow rates keep a careful
217"
-------
check to ensure that the water valve does not
shut itself off. To account for changes in
the tap water temperature, establish the *
correct flow rate daily.
NOTE 1: The relative position of the acetone
condensation level to the grid level is
critical to the successful operation of the
condensation washer. If the condensation
level is too low, the Millipore filter will
not be sufficiently removed within a
reasonable period of time and the asbestos
fibers cannot be successfully counted; if the
level is too high, excessive washing occurs
with a resulting loss of fibers and rupture of
the carbon film. As each extractor has
different characteristics, several test runs
should be made on blank Millipore-loaded grids
to determine the optimum operating conditions.
NOTE 2: It has been suggested that the rate
of acetone condensation, observed as drops
from the end of the cold finger, should be 10
drops per 30-45 seconds.
NOTE 3: A constant pressure regulator may be
required in the water line if a constant flow
cannot be otherwise attained.
8.2B.2 Vacuum Filtration Unit
Assemble the vacuum filtration unit. Place a
O.U5-ym Millipore filter on the glass frit.
Turn on the suction and center the filter on
the frit. Attach the filter funnel and turn
off suction.
8.2B.3 Sample Filtration
See 8.1.1.
8.2B.4 Sample Drying
Remove the filter funnel and place the
Millipore filter in a petri dish in an
asbestos-free oven at 45° C for at least two
hours to dry.
8.2B.5 Sampling of Filter
Using a well sharpened (1/8 inch diameter)
cork borer, cut three circular sections from
the filter. Keep one-half of the filter
undisturbed for future reference or if an
17
218
-------
ashing step is required (See 8.5). Avoid
sampling near the perimeter of the filtration
area.
8.2B.6 Grid Transfer
Pass each filter section over a static
eliminator and then place each section
particulate side down on carbon coated
specimen grids previously placed in the brass
holder. Add a 10-yl drop of acetone to each
of the grids using a micro syringe. Place the
brass holder on the cold finger of the
condensation washer which has been charged
with acetone. After the correct reflux level
has been established (8.2B.1) insert the brass
block holding the grids and check a few
minutes later to make certain that the acetone
reflux is near but below grid level. Allow
the acetone to reflux for 7-8 hours to
dissolve away the filter and leave the residue
deposited on the carbon substrate of the grid.
Turn off the heating mantle. Remove the brass
block holding the grids when no drops of
acetone can be seen falling from the cold
finger. The grids are now ready for analysis
or storage.
NOTE 1: The addition of 10 yl of acetone
directly to the filter, while recommended, may,
in the opinion of some investigators, increase
the risk of removing particulates from the
filter. There are no data available to show
it has a deleterious effect.
8.4 Electron Microscopic Examination
8.1.1 Microscope Alignment and Magnification
Calibration
Following the manufacturer's recommendations
carry out the necessary alignment procedures
for optimum specimen examination in the
electron microscope. Calibrate the routinely
used magnifications using a carbon grating
replica.
NOTE 1: Screen magnification is not
necessarily equivalent to plate magnification.
8.4.2 Grid Preparation Acceptability
After inserting the specimen into the
microscope adjust the magnification low enough
18
219
-------
(300X - 1000X) to permit viewing complete grid
squares. Inspect at least 10 grid squares for
fiber loading and distribution, debris
contamination, and carbon film continuity.
Reject the grid for counting if:
1) The grid is too heavily loaded with fibers
to perform accurate counting and diffraction
operations, A new sample preparation either
from a smaller volume of water or from a
dilution with filtered distilled water must
then be prepared.
2) The fiber distribution is noticeably
uneven. A new sample preparation is required.
3) The debris contamination is too severe to
perform accurate counting and diffraction
operations. If the debris is largely organic
the filter must be ashed and redispersed (see
8.5). If inorganic the sample must be diluted
and again prepared.
4) The majority of grid squares examined have
broken carbon films. A different grid
preparation from the same initial filtration
must be substituted.
8.4.3 Procedure for Fiber Counting
There are two methods commonly used for fiber
counting. In one method (A) 100 fibers,
contained in randomly selected fields of view,
are counted. The number of fields plus the
area of a field of view must be known when
using this method. In the other method (B),
all fibers (at least 100) in several grid
squares or 20 grid squares are counted. The
number of grid squares counted and the average
area of one grid square must be known when
using this method.
NOTE 1: The method to use is dependent upon
the fiber loading on the grid and it is left
to the judgement of the analyst to select the
optimum method. The following guidelines can
be used: If it is estimated that a grid square
(80 ym x 80 ym) contains 50-100 fibers at a
screen magnification of 20000X it is
convenient to use the field-of-view counting
method. If the estimate is less than 50, the
grid square method of counting should be
chosen. On the other hand, if the fiber count
19
220
-------
is estimated to be over 300 fibers per grid
square, a new grid containing less fibers must
be prepared (through dilution or filtration
of a smaller volume of water).
8.4.3A Field-of-View Method
After determining that a fiber count can be
obtained using this method adjust the screen
magnification to 10-20000X. Select a number
of grid squares which would be as
representative as possible of the entire
analyzable grid surface. From each of these
squares select a sufficient number of fields
of view for fiber counting. The number of
fields of view per grid square is dependent
upon the fiber loading. If more than one
field of view per grid square is selected,
scan the grid opening orthogonally in an
arbitrary pattern which prevents overlapping
of fields of view. Carry out the analysis by
counting, measuring and identifying (see
8.4.4) approximately 50 fibers on each of two
grids.
The following rules should be followed when
using the field of view method of fiber
counting. Although these rules were derived
for a circular field of view they can be
modified to apply to square or rectangular
designs.
1) count all fibers contained within the
counting area and not touching the
circumference of the circle.
2) Designate the upper right-hand quadrant as
I and number in clockwise order. Count all
fibers touching or intersecting the arc of
quadrants I or IV. Do not count fibers
touching or intersecting the arc of quadrants
II or III.
3) If a fiber intersects the arc of both
quadrants III and IV or I and II count it only
if the greater length was outside the arc of
quadrants IV and I, respectively.
U) Count fibers intersecting the arc of both
quadrants I and III but not those intersecting
the arc of both II and IV.
These rules are illustrated in Fig. 3.
20
221
-------
I
r
L 1
i
I
»
\
V
/
/
\
\
-\
III
\
Counted
Not Counted
Figure 3. Illustration of Counting Rules
for Field-of-View Method
222
21
-------
8.4.3B Grid Square Method
After determining that a fiber count can be
obtained using this method adjust the screen
magnification to 10-20000X. Position the grid
square so that scanning can be started at the
left upper corner of the grid square. While
carefully examining the grid, scan left to
right, parallel to the upper grid bar. When
the perimeter of the grid square is reached
adjust the field of view up one field width
and scan in the opposite direction. The
tilting section of the fluorescent screen may
be used conveniently as the field of view.
Examine the square until all the area has been
covered. The analysis should be carried out
by counting, measuring and identifying (see
8.4.4) approximately 50 fibers on each of two
grids or until 10 grid squares on each of two
grids have been counted. Do not count fibers
intersecting a grid bar.
8.4.4 Measurement and Identification
Measure and record the length and width of
each fiber having an aspect ratio greater than
or equal to three. Disregard obvious
biological, bacteriological fibers and diatom
fragments. Examine the morphology of each
fiber using optical viewing if necessary.
Tentatively identify, by reference to the UICC
standards, chrysotile or possible amphibole
asbestos. Attempt to obtain a diffraction
pattern of each fiber. Move the suspected
fiber image to the center of the screen and
insert a suitable selected area aperture into
the electron beam so that the fiber image, or
a portion of it, is in the illuminated area.
The size of the aperture and the portion of
the fiber should be such that particles other
than the one to be examined are excluded from
the selected area. If an incomplete
diffraction pattern is obtained move the
particle image around in the selected area to
get a clearer diffraction pattern or to
eliminate possible interferences from
neighboring particles.
Determine whether or not the fiber is
chrysotile or an amphibole by comparing the
diffraction pattern obtained to the
diffraction patterns of known standard
asbestos fibers. Confirm the tentative
identification of chrysotile and amphibole
22
223
-------
asbestos from their electron diffraction
patterns. Classify each fiber as chrysotile,
amphibole, non-asbestos, no diffraction and
ambiguous.
NOTE 1: It is convenient to use a tape
recorder during the examination of the fibers
to record all pertinent data. This
information can then be summarized on data
sheets or punched cards for subsequent
automatic data processing.
NOTE 2: Chrysotile fibers occur as single
fibrils, or in bundles. The fibrils generally
show a tubular structure with a hollow canal,
although the absence of the canal does not
rule out its identification. Amphibole
asbestos fibers usually exhibit a lath-like
structure with irregular ends, but
occasionally will resemble chrysotile in
appearance.
NOTE 3i The positive identification of
asbestos by electron diffraction requires some
judgement on the part of the analyst because
some fibers give only partial patterns.
Chrysotile shows unique prominent streaks on
the layer lines nearest the central one and a
triple set of double spots on the second layer
line. The streaks and the set of double spots
are the distinguishing characteristics of
chrysotile required for identification.
Amphibole asbestos requires a more complete
diffraction pattern to be positively
identified. As a quantitative guideline,
layer lines for amphibole, without the unique
streaks (some streaking may be present), of
chrysotile, should be present and the
arrangement of diffraction spots along the
layer lines should be consistent with the
amphibole pattern. The pattern should be
distinct enough to establish these criteria.
NOTE <*: Chrysotile and thin amphibole fibers
may undergo degradation in an electron beam;
this is particularly noticeable in small
fibers. It may exhibit a pattern for a 1-2
seconds and disappear and the analyst must be
alert to note the characteristic features.
NOTE 5: An ambiguous fiber is a fiber that
gives a partial electron diffraction pattern
resembling asbestos, but insufficient to
provide positive identification.
23
224
-------
8.ft.5 Determination of Grid Square Area
Measure the dimensions of several
representative grid squares from each batch of
grids with an optical microscope. Calculate
the average area of a grid square. This
should be done to compensate for variability
in grid square dimensions.
8.5 Ashing
Some samples contain sufficiently high levels of
organic material that an ashing step is required
before fiber identification and counting can be
carried out. If a Nuclepore filter was used for the
original preparation and if the preliminary
examination of the initial preparation shows that this
condition exists, carry out the filtration step on a
new water sample using a . 45-ym Millipore filter. If
a Millipore filter was used initially the unused half
from 8.26.5 can be ashed.
NOTE 1: A Millipore filter is specified because it is
more readily oxidized under the specified ashing
conditions.
Place the dried Millipore filter paper containing the
collected sediment into a glass vial (28 mm diameter x
80 mm high). Position the filter such that the
filtration side touches the glass wall. Place the
vial in an upright position in the low temperature
asher. Operate the asher at 50 watts (13.56 MHz)
power and 2 psi oxygen pressure. Ash the filter until
a thin film of white ash remains. The time required
is generally 6 to 8 hours. Allow the ashing chamber
to slowly reach atmospheric pressure and remove the
vial. Add 10 ml of filtered distilled water
containing 0.1 percent filtered Aerosol OT to the
vial. Place the vial in an ultrasonic bath for 1/2
hour to disperse the ash. Dilute the sample if
required.
Assemble the 25 mm diameter filtering apparatus. (See
Note 1) Center a 25 mm diameter O.U5-ym Millipore or
.1-ym Nuclepore filter (with the 2-pm Millipore
backing) on the glass frit. Apply suction and
recenter the filter if necessary. Attach the filter
funnel and turn off the suction. Add the water
containing the dispersed ash from the vial to the
filter funnel. Apply suction and filter the sample.
After drying this filter it is ready to be used in
preparing sample grids as in 8.2A or 8.2B.
24
225
-------
NOTE 2; In specifying a 25-mm diameter filter it is
assumed that the ashing step is necessary mainly
because of the presence of organic material and that
the smaller filtering area is desirable from the point
of view of concentrating the fibers. If the sample
contains mostly inorganic debris such that the smaller
filtering area will result in over-loading the filter,
the 47-mm diameter filter should be used.
NOTE 3: It will be noted that a 10-ml volume is
filtered in this case instead of the minimum 50-ml
volume specified in 8.1.1. These volumes are
consistent when it is considered that there is
approximately a 5-fold difference in effective
filtration area between the 25-mm diameter and 47-mm
diameter filters.
8.6 Determination of Blank Level
Carry out a blank determination with each batch of
samples prepared, but a minimum of one per week.
Filter a fresh supply (500 ml) of distilled, deionized
water through a clean ,1-ym membrane filter. Using
the selected filter type, filter 200 ml of this water,
prepare the electron microscope grid, and count
exactly as in the procedures 8.1 - 8.U. Examine 20
grid squares and record this number of fibers. A
maximum of two fibers in 20 grid squares is acceptable
for the blank sample.
NOTE 1: The monitoring of the background level of
asbestos is an integral part of the procedure. Upon
initiating asbestos analytical work, blank samples
must be run to establish the initial suitability of
the laboratory environment, cleaning procedures, and
reagents for carrying out asbestos analyses.
Analytical determinations of asbestos can be carried
out only after an acceptably low level of
contamination has been established.
9. Calculations
9.1 Fiber Concentrations
Grid Square Counting Method - If the Grid Square
Method of counting is employed, use the following
formula to calculate the total asbestos fiber
concentration in MFL.
C = (F x Af) / (G x A x VQ x 1000)
If ashing is involved use the same formula but
substituting the effective filtration area of the 25-
mm diameter filter for A^ instead of that for the 47-
25
226
-------
mm diameter filter. If one-half the filter is ashed,
multiple C by two.
C = Fiber concentration (MFL)
F * number of fibers identified in "G" grid
squares
Af * effective filtration area of filter paper
(mm*) used in grid preparation used for fiber
counting
Ag » Average area of one grid square (mm2)
G = number of grid squares analyzed
Vo = original volume of sample filtered (ml)
Field-of-View Counting Method - If the Field-of-View
Method of counting is employed use the following
formula to calculate the total asbestos fiber
concentrations (MFL)
C = (F x Af x 1000) / (Ay x Z x VQ)
If ashing is involved use the same formula but
substituting the effective filtration area of the 25-
mm diameter filter for Af instead of that for the 47-
mm diameter filter.
C = fiber concentration (MFL)
F = number of fibers identified in area
examined (Ay x Z)
Af = effective filtration area of filter paper
(mm2) used in grid preparation for fiber
counting
Ay = area of one field of view (
Z = number of fields of view examined
VQ = original volume of sample filtered (ml)
9.2 Estimated Mass Concentration
Calculate the mass ftig) of each fiber counted using
the following formula:
M=LxW«xDx 10-*
If the fiber content is predominantly chrysotile, the
following formula may be used:
26
227
-------
M = xLxW«xDx 10-*
where M * mass (yg)
L * length (urn)
W » width (urn)
D » density of fibers (g/cm3)
Then calculate the mass concentration (yg/1) employing
the following formula.
Mc * C X Mf x 10*
where Mc = mass concentration (yg/1)
C = fiber concentration (MFL)
M.f = mean mass per fiber (yg)
To calculate Mf use the following formula:
n
fff - 2lMi/n
i - 1
where M = mass of each fiber, respectively
n * number of fibers counted
NOTE 1: Because many of the amphibole fibers are lath
shaped rather than square in cross section the
computed mass will tend to be high since laths will in
general tend to lie flat rather than on edge.
NOTE 2: Assume the following densities: Chrysotile
2.5, Amphibole 3.25
9.3 Aspect Patio
The aspect ratio for each fiber is calculated ty
dividing the length by the width,
10. Reporting
10,1 Report the following concentration as MFL
a. Total fibers
b. Chrysotile
c. Amphibole
27
228
-------
10.2 Use two significant figures for concentrations greater
than 1 MFL, and one significant figure for
concentrations less than 1 MPL.
10.3 Tabulate the size distribution, length and width.
10.4 Tabulate the aspect ratio distribution.
10.5 Report the calculated mass as yg/1.
10,6 Indicate the detection limit in MFL.
10.7 Indicate if less than five fibers were counted.
10.8 Include remarks concerning pertinent observations,
(clumping, amount of organic matter, debris) amount of
suspected though not identifiable as asbestos
(ambiguous) .
11. Precision
11.1 Intra Laboratory
The precision that is obtained within an individual
laboratory is dependent upon the number of fibers
counted. If 100 fibers are counted and the loading is
at least 3.5 fibers/grid square, computer modeling of
the counting procedure shows a relative standard
deviation of about 10% can be expected.
In actual practice some degradation from this
precision will be observed but should not exceed ± 15%
if several grids are prepared from the same filtered
sample. The relative standard deviation of analyses
of the same water sample in the same laboratory will
increase due to sample preparation errors and a
relative standard deviation of about ± 25 - 30% will
occur. As the number of fibers counted decreases, the
precision will also decrease approximately
proportional to /N where N is the number of fibers
counted.
11.2 Inter Laboratory
While there have been numerous inter laboratory
testing programs, there have been few carried out
using the same procedure. Those that have been done
indicate that agreement within a factor of two is
achieved if 100 fibers can be counted.
229
-------
12, Accuracy
12.1 Fiber concentrations
As no standard reference materials are available, only
approximate estimates of the accuracy of the procedure
can be made. At 1 MFL, it is estimated that the
results should be within a factor of 10 of the actual
asbestos fiber content.
This method requires the positive identification of a
fiber to be asbestos as a means for its quantitative
determination. As the state-of-the art precludes the
positive identification of all of the asbestos fibers
present, the results by this method, as expressed as
MFL, will be biased on the low side and assuming no
fiber loss represent .2 - .6 of the total asbestos
fibers present.
12.2 Mass concentrations
As in the case of the fiber concentrations, no
standard samples of the size distribution found in
water are available. The accuracy of the mass
determination should be somewhat better than the fiber
determination because a larger fraction of the large
fibers, which contribute the major portion of the
mass, are identifiable. This will reduce the bias of
low results due to difficulties in identification. At
the same time, the assumption that the thickness of
the fiber equals the width will result in a positive
error in determining the volume of the fiber and thus
give high results for the mass.
SELECTED BIBLIOGRAPHY
Seaman, D. R. and D. M. File. Quantitative Determination of
Asbestos Fiber Concentrations. Anal. Chem. 48(1): 101-110, 1976.
Lishka, R. J., J. R. Millette, and E. F. McFarren. Asbestos
Analysis by Electron Microscope. Proc. AWWA Water Quality Tech.
Conf. American Water Works Assoc., Denver, Colorado XIV - 1 - XIV
- 12, 1975.
Millette, J.R. and E. F. McFarren. EDS of Waterborne Asbestos
Fibers in TEM, SEM and STEM. Scanning Electron Microscopy/1976
(Part III) 451-460, 1976.
Cook, P. M., I. B. Rubin, C. J. Maggiore, and W. J. Nicholson.
X-Ray Diffraction and Electron Beam Analysis of Asbestiform
Minerals in Lake Superior Waters. Proc. Inter. Conf. on Environ.
Sensing and Assessment 34(2): 1-9, 1976.
McCrone, W. C. and I. M. Stewart, Asbestos. Amer. Lab. 6(4):10-
18, 1974.
230 "
-------
Mueller, P. K., A. E» Alcocer, R. L. Stanley, and G. R. Smith,
Asbestos Fiber Atlas. U.S. Environmental Protection Agency
Technology Series, EPA 650/2-75-036, 1975.
Glass, R. w., improved Methodology for Determination of Asbestos
as a Water Pollutant. Ontario Research Foundation Report, April
30, 1976. Mississauga, Ontario, Canada.
30
231
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Analytical Methodology for the Determination of Asbestos
by Transmission Electron Microscopy
The analytical procedure used by Walter C. McCrone Associates,
Inc., for the determination of asbestos in environmental samples is substan-
tially similar to that given in the U.S. EPA "Preliminary Interim Procedure
for Determining Fibrous Asbestos". * Although this procedure was written
for water samples, the techniques for preparation of the filter for examination
and the criteria for the identification of the asbestiform minerals are equally
applicable to air samples. Details of the procedure follow.
Working in a laminar flow clean bench (see attached laboratory
description), discs approximately 3 mm in diameter are punched out of the
filter. These discs are then placed face-down on previously carbon-coated
electron microscope support grids either of copper, if only chrysotile is
expected, or nylon. Nylon is used for samples in which there is a reason-
able likelihood of amphibole fibers in order that chemical analyses may be
performed on the fibers, by either the X-ray energy or wavelength dispersive
system fitted to the microscope. The use of nylon minimizes extraneous
X-ray signals from the support grid which would otherwise saturate the
detector system. Such an analysis is essential in order to classify the amphi-
bole type present. The grids are then transferred to a cold finger in a Soxhiet
extraction apparatus in which the membrane filter is dissolved using acetone
for Millipore Type MF and for Gelman GN-6 Metricel filters or chloroform
for Nuclepore filters. A "wicking" method may also be used for Nuclepore
filters but is unsuitable for the Millipore or Gelman types. Previous work
has shown us that there is very little risk of contamination in transferring
the filter on the electron microscope grid to the Soxhiet extractor. Further-
*Available from U.S. EPA Environmental Research Laboratory, Athens,
GA 30601.
232
waiter c. me crone associates, inc.
-------
more, by dissolving the filter in situ on the grid ("direct transfer"), the
risk of losing portions of the sample is minimal., Techniques involving
transfer of a liquid suspension directly to the electron microscope grid
are more subject to error since there is frequently a size separation as
the meniscus of the drying drop recedes. Procedures involving "rub-out"
techniques, though of some value in obtaining mass concentration data are
not applicable to fiber number or size distribution determinations as they
intentionally degrade the fibers to unit fibrils thus altering their size and
simultaneously increasing their numbers.
The sample grids are examined on the electron microscope
1 2
(JEM 200 * or EMMA 4* ) using a magnification such that the intermediate
lens aperture is in focus in the specimen plane. It is thus possible, by
inserting the aperture and switching to the diffraction position, to obtain a
selected area electron diffraction (SAED) pattern of the fiber with no other
adjustments to the microscope. In this way it is possible to spot check the
diffraction pattern, of individual fibers very rapidly. The JEM 200 is used on
those samples in which only chrysotile is of interest. EMMA 4, with the
capability for X-ray fluorescence analysis of individual fibers, is used where
the identification of amphibole types present is required. Both instruments
have a selected area electron diffraction capability.
Prior to commencing measurement the electron microscope
grid is scanned at a low magnification, approximately 2000-4000X to ensure
uniformity of dispersion on the filter. In the case of non-uniform deposition,
which may occur for example with cemented or aggregated fibers, several
grids may be examined from the same filter. This prior examination indicates
to the analyst which areas should be examined to obtain a truly representative
analysis of the sample. Magnifications in excess of 10,000 X are required for
*1 JEM 200. 200 Kv transmission electron microscope manufactured by Japan
Electron Optics Laboratories (JEOL)
*2 EMMA 4. Combined 100 Kv transmission Electron Microscope-Micro-
probe Analyzer manufactured by Associated Electrical Industries (AJEI)
233 waiter c. me crone associates, inc,
-------
the observation of the smallest chrysotile fibrils present.
The magnification of examination used in the JEM 200 is
14, 600 on the viewing screen; that used in EMMA 4 is 24,800. As stated
above, these magnifications are based on user convenience in switching from
viewing to diffraction.
The length and width of each asbestos fiber is recorded. Only
fibers which are positively identified as asbestos are measured. Interpolation
from intervals scribed on the viewing screen allows an accuracy of measure-
ment on the screen of approximately 0. 05 cm. This corresponds to an
accuracy in size measurement of about 0.02-0.04 pm. Measurements of the
individual fibers are computer processed to give listings of the length and
width of the fibers, together with a computed mass of each fiber computed on
7
the basis of density, D, and dimensions, L and W (D x L x W ). A value of
3.3 is taken as the mean density of amphibole fibers: a density of 2.3 is used
for chrysotile. Because many of the amphiboles are lath-shaped rather than
square in cross section, this figure may well be slightly high, since the laths
will, in general, tend to lie flat rather than on edge. There is, however, a
finite possibility that some laths will be on edge and, due to the very small
size of many of the fibers of interest, the approximation to a square fiber will
not give more than a slightly high bias to the mass readings. The program
automatically assigns the longest dimension to the fiber length and excludes all
particles with an aspect ratio below three.
Also presented in the computer printout are the calculated num-
ber of fibers per unit volume, the calculated mass of fiber per unit volume,
the size distribution of the fibers based on length and width, and the distribu-
tion of fibers by aspect ratio together with the relevant statistical information
on these parameters. A physical description of the sample accompanies the
measurements and is considered an integral and essential part of the analysis.
A sample of a complete analysis, description and computer printout is attached.
234
waiter c. me crone associates, inc.
-------
Sample
Much chunky and chiplike material ranging to quite large sizes
and showing a wide range of composition — Mg, Si and some Fe, mainly Si,
mainly Ca or Fe-Si types — although the Mg-Si type predominates. Additionally,
fine agglomerate material, organic matter, some spherical inorganic particles
and chrysotile are found,in this sample. The spheroidal particles are found
normally, to be Al-Si-Fe composition but wide ranging variation does occur.
Antigorite laths (e.g. fibers) noted. Occasional";massive" fibers are present or
fibers which are lathlike. These are found to contain Mg-Si-Ca-Fe and some S,
consistently.
• -..'iSiap^g^fwi^spc^sa'Js
Probe of large chunky material
Probe of large chunky material
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t '
' * . - -6
PRESERVATION OF PHE1IOLIC COMPOUNDS
IN WASTEWATERS
Mark J. Carter * and Madeliene T. Huston
Central Regional Laboratory
Environmental Protection Agency
1819 W, Pershing Road
Chicago, Illinois 60609
240
-------
BRIEF
The combination of strong acid or base with sample storage at 4°C stabilized
phenolic compounds in wastewaters for at least 3-4 weeks. There is a positive
relationship between microbiological activity and chemical stability of the
samples studied.
ABSTRACT
Copper sulfate and phosphoric acid with sample-storage at 4°C is a common
preservative technique used for phenolic compounds in wastewaters. However,
there are no data showing its effectiveness. A study was conducted to compare
the preservation method with the addition of strong base or acid and sample
storage at 25° and 4°C. The addition of 1 ml cone H.SO./l with sample storage
at 4°C was most consistently effective in preserving stability for 3-4 weeks.
However, the other chemical preservatives were found to be effective for at
least 8 days. Substantial loss of phenolic compounds rapidly occured in all
samples unless the chemical preservative was added immediately after sample
collection. A positive correlation was found between loss of phenolic compounds
and microbiological activity suggesting the latter was the dominant factor in
determining sample stability.
INTRODUCTION
The 1972 Amendments to the Clean Water Act have resulted in limitations on
the concentration and loading of pollutants that can be discharged by industries
and municipalities (1). The need to monitor these discharges has substantially
increased the number of environmental samples requiring analysis, especially for
toxic substances such as phenolic compounds.
*
Xelly has reviewed the literature for methods of analyses of phenolic compounds
in wastewaters (2). The most common methods are the 4-aminoantipyrine (4-AAP)
(3-6) and 3-methyl-2-faenzothiazolinone hydrazone (MBTH) (7,8) colorimetric
procedures and the ultraviolet bathochromic shift method (9). The difficulty and
equipment requirements for these methods often results in samples being shipped
241
-------
Chemical Analysis of Samples. The first analysis of each sample was completed
within two hours of sample collection. All samples, standards and blanks were
distilled from acidic solution, to separate phenolic compounds from potential
interferences (13). The distillates were analyzed by an automated version of the
4-aminoantipyrine method shown in figure 1.
The buffered potassium ferricyanide reagent was prepared by adding 2.0 g
potassium ferricyanide, 2.1 g boric acid, 3.75 g potassium chloride, 44 ml of
1 S sodium hydroxide and 0.5 ml Brij-35 (Technicon Corp. No. T21-011Q) to a
volumetric flask and diluting to 1 1. The 4-aminoantipyrine reagent was preparad
by diluting 0.65 g of the chemical to 1 1. Both reagents were filtered through
a 0.45 im membrane filter before use.
Two control standards were prepared and preserved with copper sulfate and
phosphoric acid by an independent analyst at the beginning of each study to
checJc on the consistency of the day-to-day standard preparation and instrument
calibration.
Standard Plate Count. Plate count agar was prepared fresh just before use, added
to petri dishes and 1, 0.1 and 0.01 ml of each sample was plated in triplicate
(13). All samples were incubated at 35°C for 24 hrs. Only those plates having
30-300 colonies were considered valid and the values reported in Table I are an
average of the three replicate dilutions.
RESULTS AND DISCUSSION
The stability of phenolic compounds in ncn-wastewaters aqueous solutions has
been studied by several investigators. Phenolic compounds are good preservatives
at high concentrations (>0.5%) but are readily biodegraded at lower concentrations
(14-17). Chambers and Kabler found no detectable nonbiological degradation (15).
Extremes in pH (18-22), temperature (23-26) and the use of toxic chemicals (27-29)
have been used to reduce microbiological activity in aqueous solution. Strong base
(4,30,31), acid (4) and copper sulfate-phosphoric acid (11,12) in combination
with temperature control have been used to stabilize phenolic compounds in surface
waters.
- 3 -
243
-------
to a large centralized laboratory for analysis. The shipping process can lead
to substantial delays between sample collection and analysis. As a result, the
effectiveness of the sample preservation technique will substantially affect the
accuracy of the data. It is also necessary to consider sample stability during
the collection process, especially if the 24-hour composite method is used (10) .
Very little data exist in. the literature to give guidance about the best method
to stabilize phenolic compounds in wastewaters. The current recommended technique
(11) is a modification of work performed over thirty years ago (12) . Ettinger,
et al., found that copper sulfate effectively preserved river water and river
water seeded with sewage for two to four days when the samples were stored at
25°C. Subsequent to the work of Sttinger, phosphoric acid was added to the copper
sulfate preservative to keep the metal ions in solution when added to alkaline
samples (11). In addition, it was recommended that all samples be stored at
4°C until analysis.
No data was presented until 1974 about the effectiveness of the combined
phosphoric acid, copper sulfate, 4CC storage technique for preserving phenolic
compounds in water samples. Afghan, et al., showed that either strong acid
or base has more effective in retarding bacterial activity and stabilizing phenol
in Great Lakes' waters than the combined copper sulfate preservative (4).
•
The observation of Afghan raises doubt that the copper sulfate - phosphoric acid
preservation method is best for stabilizing phenolic compounds in wastewaters.
Therefore, a study was undertaken to determine the most effective and practical
preservation method and maximum allowable holding time for phenolic compounds in
wastewaters.
METHODS
Preparation of Samples. Fresh samples were collected in 5 gal high density
polyethylene jugs and immediately brought to the laboratory. The water samples
were homogenized with a Tekmar Super. Dispax system, preserved and split into 250 ml
high density polyethylene bottles. Some samples were spiked with phenol to raise
the starting concentration to a level that could be accurately measured. The
samples were preserved and stored as described in figures 2-5.
- •> 242 e
• .. \ *
-------
Chemical Analysis of Samples. The first analysis of each sample was completed
within two hours of sample collection. All samples, standards and blanks were
distilled from acidic solution, to separate phenolic compounds from potential
interferences (13). The distillates were analyzed by an automated version of the
4-aminoantipyrine method shown in figure 1.
The buffered potassium ferricyanide reagent was prepared by adding 2.0 g
potassium ferricyanide, 2.1 g boric acid, 3.75 g potassium chloride, 44 ml of
1 N sodium hydroxide and 0.5 ml Brij-35 (Technicon Corp. No. T21-0110) to a
volumetric flask and diluting to 1 1. The 4-aminoantipyrine reagent was preparad
by diluting 0.65 g of the chemical to 1 1. Both reagents were filtered through
a 0.45 yjn membrane filter before use.
Two control standards were prepared and preserved with copper sulfate and
phosphoric acid by an independent analyst at the beginning of each study to
check on the consistency of the day-to-day standard preparation and instrument
calibration.
Standard Plate Count. Plate count agar was prepared fresh just before use, added
to petri dishes and 1, 0.1 and 0.01 ml of each sample was plated in triplicate
(13). All samples were incubated at 354C for 24 hrs. Only those plates having
30-300 colonies were considered valid and the values reported in Table I are an
average of the three replicate dilutions.
RESULTS AND DISCUSSION
The stability of phenolic compounds in ncn-wastewaters aqueous solutions has
been studied by several investigators. Phenolic compounds are good preservatives
at high concentrations (>0.5%) but are readily biodegraded at lower concentrations
(14-17). Chambers and Kabler found no detectable nonbiological degredation (15).
Extremes in pH (18-22), temperature (23-26) and the use of toxic chemicals (27-29)
have been used to reduce microbiological activity in aqueous solution. Strong base
(4,30,31), acid (4) and copper sulfate-phosphoric acid (11,12) in combination
with temperature control have been used to stabilize phenolic compounds in surface
waters.
- 3 -
243
-------
The stability of phenolics in three different wastewaters preserved with copper
sulfate - phosphoric acid and stored at 4°C was studied first. The results arc
shown in Figure 2. The raw sewage was fairly weak with a biochemical oxygen
demand (BOO) of only 95 mg/1 and the treated sewage sample was collected after
secondary biological treatment but before chlorination. The industrial wastewater
was collected from the Grand Calumet River which is essentially a composite from
the South Chicago, industrial area.
Since the samples chosen for study often had low background concentrations of
phenolics, each was spiked with phenol as needed so that changes in phenolic
concentration wovld be easier to determine. Phenol was chosen for the spike
because Kaplin, et al., found phenol to be the least stable of all the phenolic
compounds in natural waters (32-35).
The most important result of study 1 was the rapid loss of phenolics from the
samples at 4°C with no addition of any chemical preservative. The percentage
loss of phenolics within 24 hrs. for the industrial waste, raw and treated
sewage saoples was 35, 80 and 40%, respectively. Ettinger, et al., reported that
an unpreserved river water sample stored at 2°C lost only 15% of the original
phenolic content after 4 days (12). However/ the same sample stored at 25°C lost
all of its phenolic compounds within 2 days. The results from these two studies
show that the loss of phenolics from unpreserved samples is variable depending
upon sample type but significant in all cases. The expected precision of sample
analysis was determined from daily analysis of the control A and 3 samples to be
± 12 vg/1 (20). There was no statistically significant change in phenolic
concentration over the 22 day study for the samples preserved with copper sulfara-
phosphoric acid and stored at 4°C. However, there were large day-to-day changes
in the concentration of phenolics measured. This poor precision was determined
to be caused by the problem in taking a representative sample for analysis due
to the presence of particulate matter. The problem was solved in later studies
by homogenizing all samples before analysis.
In the second study (figure 3), the effectiveness of the combined copper sulfate-
phosphoric acid preservative was sutdied vs. sample type. Activated sludge was
added to raw sewage to create a sample that was organically rich and biologically
active. This sample was stable for 12 days, but degraded to 85% of the original
244
— 4 -
-------
phenol concentration after 33 days. The other samples were stable for the duration
of the study. The dip in all values on day 1 of the study was attributed to
improper calibration.
Baylis reported the use of 1.2 ml 1 M NaOH/1 of sample to preserve phenolic
compounds in potable water samples(30). However, Ettinger, et al., found Baylis1
procedure to be ineffective for sewage seeded stream samples (12). Kaplin and
Frenko found that a hundred-fold increase in base concentration was effective for
preserving stream waters (31). Afghan, et al., verified the effectiveness of the
higher concentration of base for preserving lake waters (4). Afghan
also showed that 0.1 M HCl was an effective preservative.
The effectiveness of strong base or acid in preserving phenolic compounds was
compared with copper sulfate in the third study (figure 4). The concentration
of phenolic compounds was stable in the raw sewage sample studied when stored at
4°C regardless of the preservative used. However, the sulfuric acid and copper
sulfate preserved samples deteriorated rapidly after eight and two days, respective
when stored at 25°C.
Doetsch and Cook reported that a common feature of acidophilic bacteria was a
resistance to copper ions (23). Growth of acidophilic bacteria occurs at pH 2-5
which is the pH range for the copper sulfate preservative. These facts makes the
use of copper sulfate at pH 4 suspect as a good preservative, especially if the
samples are not stored at 4aC. The same sample with 2 ml cone H.SO./l - which
produces a pH cf about 1.5 - at 2S°C was stable for eight days. Kushner has
reported far fewer microorganisms can tolerate pH 1.5 than 4 (22). It is
interesting to note that even at pH 1.5 but at 25°C that the phenolic concentration
decreased substantially. This observation indicates that, while neither acidifi-
cation or cold storage stabilizes phenolic compounds in a wastewater, the
combination does.
In order to evaluate the biological induced degredation of phenolic compounds,
microbiological activity was measured on a raw and secondary treated sewage.
Samples were preserved as indicated in Table I and total plate counts taken after
1 hr. (day 0), 8 and 20 days. The only secondary sewage aliguot that showed any
significant activity was the chemically unpreserved sample stored at 4°C. The
-5-245
-------
microbiological activity noted corresponds very closely with the chemical stability
of phenolics in treated sewage found in studies 1 and 2.
i
The unpreserved raw sewage sample stored at 4°C showed very great microbiological
activity which corresponds to the phenolic instability noted in Study 1. The
addition of 2 ml cone H.SO /I initially reduced the microbiological activity
significantly. "However, by day eight, the activity increased five-fold and then
decreased slightly again by day twenty. This trend corresponds closely with the
rapid loss of phenolics in Study 3, after day eight and then a moderate but
continued loss thereafter. The same sample stored at 4°C with 2 ml cone H.SO /I
showed at least a ten-fold lower microbiological activity and a corresponding
increase in chemical stability shown in Study 3.
The raw sewage sample with copper sulfate-phosphoric acid and stored at 4°C
exhibited greater microbiological activity than the aliquot with sulfuric acid.
This observation corresponds to the moderate effectiveness of this preservative -
stability in studies 1 and 3 and instability of the raw sewage in Study 2. Addition
of strong caustic also lowered the microbiological activity of the raw sewage
sample. However, the higher concentration of caustic, 10 ml 10 N NaOH/1, was
required for a quick initial kill. The high initial microbiological activity
of the 2 ml 10 N NaOH/1 aliquot did not affect the chemical stability found in
Study 3.
•
Increasing the concentration of H _SO . two-fold with storage at 2S°C, reduced the
microbiological activity to the same level as the aliquot stored at 4°C with
2 ml cone H.SO./l. A fourth study was conducted to detemine if a greater acid .
concentration could preserve phenolic stability without cold storage. The results
in Figure 5, show good stability for the aliguots preserved with 2 ml H SO /I at
4°C and 4 ml H2SO4/1 at 25°C. The aliquot with 2 ml H SO4/1 at 2S°C showed a
substantial loss of phenolic compounds after the eighth day.
The enhanced stability of the samples preserved with the higher acid concentration
is- excellent evidence that the greatest cause of sample instability is caused by
microbiological and not chemical acitivity. Gordon claimed that phenolic compounds
in many refinery effluent waters can be oxidized in acid solution (35). However,
246
-------
he did not state the temperature conditions for storage or provide any data to
support the claim. Emerson noted that phenol was less reactive under oxidizing
acidic than basic conditions (37). Stewart'(38) and Waters (39) also noted that
phenolic compounds were more reactive in basic solution. However, in practice
basic preservation does not cause instability of phenolic compounds especially
if stored at 48C (4,30,31,36).
CONCLUSIONS AND RECOMMENDATIONS
All samples quickly lost phenolic compounds in the absence of a chemical
preservative, even if stored at 4°C. Therefore, all samples must be chemically
preserved at the'time of collection. The chemical preservative must be added to
the first aliquot of a composite sample.
The desired time period for holding samples determines the choice of chemical
preservatives. All preservatives studied, NaOH, 3,30 and CuSO. - H PO , were
effective - no more than 5% phenolic compound loss - for at least 12 days whan
the samples were stored at 4°C. Strong base or acid were effective for 26 and
28 days, respectively, when the samples were stored at 4aC.
The use of acid or base preservation has the advantage of eliminating the use of
one separately preserved bottle specifically for phenolics analysis (11,13). The
choice of acid or base preservation will depend on whether cyanide (base preserved)
or nutrient (acid preserved) analyses will be performed. The advantage of acid
preservation is that sulfides, a common interference in the colorinetrie methods,
will be driven out of the sample (35). The basic preservation will be advantageous
if any organic extraction is required in the analysis method to remove organic
interferences (13).
*
Use of 2 ml cone H.SO./l with sample storage at 4°C is recommended over the use
Of 4 ml cone H_SO /I at 25°C. The former conditions combine the preservative
Dualities of low temperature and pH and are milder conditions chemically which
should reduce the possibility of undesirable chemical reactions.
- 7 -'
247
-------
ACKNOWLEDGEMENT
The authors would like to thank M. Anderson, C. Steiner and 0. Grothe, EPA,
Chicago for .their assistance with the microbiological work and data interpretation.
Mention of trade names or commercial products does not- imply endorsement by
the Environmental Protection Agency or the Central Regional Laboratory.
8 -
248
-------
LITERATURE CITED
I
1. Amendment to the Federal Water Pollution Control Act, Public Law 92-500,
October 18, 1972.
2. Kelly, J.A., "Determination of Phenolic - Type Compounds in Water and
Industrial Wastewaters," Oklahoma State Univ., Stillwater, Okla., NTIS
* ORO-4254-11, 1972.
3. Mohler, E.F., Jr. and Jacob, L.N., Anal. Chem., 29, 1369 (1957).
4. Afghan, B.K., Belliveau, P.E., Larose, R.H. and Ryan, J.F., Anal. Chim. Acta,
JU, 355 (1974) .
5. Ettinger, M.S., Ruchhoft, C.C. and Lishka, R.J., Anal.Chem., 23_, 1783 (1951).
6. Gales, M.E., Jr. and Booth, R.I., Journal AHWA, 63, 540 (1976).
7. Friestad, J.O., Ott, D.E. and Gunther, F.A., Anal.Chem., 41, 1750 (1969).
3. Goulden, P.O., Brooksbank, P. and Day, M.B., Anal.Chem., 45, 2430 (1973).
9. Fountaine, J.E., Joshipura, P.B., Keliher, P.N. and Johnson, J.D., Anal.Chan.,
46_, .62 (1974).
10. "Handbook for Monitoring Industrial Wastewater," U.S. Environmental Protection
Agency, Technology Transfer, Cincinnati, CH, 1973.
11. "Manual of Methods for Chemical Analysis of Water and Wastes," U.S. Environments
Protection Agency, Technology Transfer, Cincinnati, OH, 1974.
12. Ettinger, M.S., Schott, S. and Ruchhoft, C.C., Journal AWWA, 35, 299 (1943).
13. "Standard Methods for the Examination of Water and Wastewater," 14th ed.,
American Public Health Association, Washington, D.C., 1976.
14. Harlow, I.P., Ind. Sng. Chem., 3_1_, 1346 (1939).
15. Chambers', C.W. and Kobler, P.W., in "Developments in Industrial Microbiology,"
Vol. 5, American Institute of Biological Sciences, Washington, D.C., 1964.
16. Erikson, D., Jour. Bact., 41, 277 (1941).
17. ZoBell, C.E. and Brown, B.F., J. Mar. Res., 5_, 178 (1944).
18. Ruchhoft, C.C., Ettinger/ M.B. and Walker, W.W., Ind. Eng. Chem., 32, 1394 (1949)
19. Hewitt, L.F., in "Microbial Ecology," University Press, Cambridge, England, 1957.
- 9 -
249
-------
20. Wood, E.J. Ferguson, "Microbiology of Oceans and Estuaries," Elsevier
Publishing Co., New York, N.Y., 1967.
21. Weiss, R.L., Liranol. and Oceanogr., 18, 877 (1973).
22. Kashner, D.J., in "Inhibition and Destruction of the Microbial Cell,"
W.B. Hugo, ed., Academic Press, London, 1971.
23. Waksman, S.A. and Carey, C.L., Jour. Bact., 29, 531 (1935).
24. ibid, p. 545.
25. Butterfield, C.T., Sew. Works Jour. > 5_, 600 (1933).
26. Olsen, R.H. and Metcalf, E.S., Science, 162., 288 (1968).
27. Ruchhoft, C.C. and Placak, Q.S., Sew. Works Jour., 14, 638 (1942).
28. Doetsch, R.N. and Cook, T.M., "Introduction to Bacteria and their Ecobiology,"
Oniversity Park Press, Baltimore, Md., 1973.
29. Pelczar, M.J., Jr. and Reid, R.D,, "Microbiology," 2nd ed., McGraw-Hill- Sook Co.
New York, N.Y., 1965.
30. Baylis, J.R., Water Works and Sewerage, 79, 341 (1932).
31. Kaplin, V.T. and Frenko, N.G., Gig. Sanit., 26, 68 (1961).
32. Kaplin, V.T., Fesenko, H.G., Babeshkina, 2.M. and Simirenko, V.I.,
Gidrokhim. Materialy, 37_, 158 (1964). Chea.Abs. , 62_, 14332.
33. Kaplin, V.T., Panchenko, S.S. and Fesenko, N.G., Gidrokhim.Materialy, 40,
134 (1965). Chem.Abs., 64, 13906.
34, Kaplin, 7.T., Semenchenko, L.V. and Ivanov, E.G., Gidrokhin,Materialy, 46,
199 (1968). Chem.Abs. 69, 69563.
35. Kaplin, V.T., Panchenko, S.E. and Fesenko, N.G. Gidrokhim.Matarialy, 42,
262 (1966). Chem.Abs., 67, 57105.
36. Gordon, G.E., Anal.Chem. , 3_2_, 1325 (1960).
37. Emerson, E., Jour. Orgr. Cham. , 8_, 417 (1943) .
38. Stewar-t, R., "Oxidation Mechanisms, Applications to Organic Chemistry,"
W.A. Benjamin, Inc., N.Y., N.Y., 1964.
39. Waters, W.A., "Mechanisms of Oxidation of Organic Compounds," John Wiley & Sons,
Inc., N.Y., N.Y., 1964.
10 -
250
-------
Table I. Effectiveness of Preservatives in Sterilizing Sewage as
Indicated by Total Plate Counts
Preservation Method
Raw Sewage
4°C
2 ml cone H,SO., 2S°C
2 4
2 ml cone H_SO., 4°C
2 4
4 ml cone H-SO,, 25eC
2 4
CttSO., H.PO., 4°C
4 34
2 ml ION NaOH, 4°C
10 ml ION NaOH, 4°C
Total Plate County, Colonies/ml
Day 0
Day 8
Day 20
>»30,000
730
560
6,300
28,000
230
>»30,000
3,500
70
40
800
110
90
>»30,000
2,200
200
b
600
270
100
Secondary Treated Sewage Before Chlorination
4°C . 23,000
<30
2 ml cone H_SO., 25°C
2 4
2 ml cone H.SO., 4°C
2 4
4 ml cone H.SO., 2S"c
2 4
CaS04/ H3P04, 4°C
2 ml ION NaOH, 4aC
10 ml ION NaOH, 4°C
<30
<30
40
<30
20,000 5,400
<30 <30
<30 <30
<30 <30
<30 <30
<30 <30
<30 <30
Volume of acid or base added per liter of sample. Copper sulfate, phosphoric
acid preservative prepared as described in ref. 13. Temperatures refer to storage
conditions. .
b
Confluent colonies
Elated within 1 hr. of preservation
- 11 -
251
-------
Figure 1. Automated phenol manifold diagram. Numbers in parentheses
correspond to the flow rate of the pumptubes in ml/min. Numbers
adjacent to glass coils and fittings are Technicon Corp. part numbers
Figure 2. Plot of stability of phenolic compounds in several wastcwaters with
time; Study 1. All samples with points plotted as "B" were preserved
with l.Og CuSO. • 5 H 0/1, the pH brought to 4.0 with phosphoric acid
and then stored at 4°C. Samples plotted as "A" were stored at 4°C
with no chemical preservatives. Both industrial waste, raw and
treated sewage samples were spiked with phenol to bring their initial
concentrations to 50, 100 and 60 wg/l» respectively.
Figure 3. Plot of stability of phenolic compounds in several wastewaters with
time; Study 2. 'All samples were stored at 4°C. The industrial
waste, raw and treated sewage samples were spiked with phenol to
bring their initial concentrations to 110, 165 and 110 yg/1^ respecti
Figure 4. Plot of stability of phenolic compounds in a raw sewage sample presex
with several chemicals; Study 3, Aliquots 1 and 2 were preserved wit
copper sulfate and phosphoric acid and stored at 25 and 4°C, respecti
Aliquots 3 and 4 were preserved with 2ml cone H.SO./l and stored at
25 and 4°C, respectively. Aliquot 5 was preserved with 2 ml 10 N
NaOH/1 and stored at 4°C, Aliquots 1-4 were spiked with phenol to
bring their initial concentrations to 125 ug/1. Aliquot 5 was spikec
with phenol to bring its initial concentration to 130 ug/1.
Figure 5. Plot of stability of phenolic compounds in a raw sewage sample
preserved with several concentrations of sulfuric acid. Aliquots
3 and 5 were preserved with 2 ml cone H-SO./l and stored at 25 and
4°C, respectively. Aliquot 4 was preserved with 4 ml cone H_SC./1
and stored at 25°C. All samples were spiked with phenol to cring
their initial concentration to 130 yg/1.
- 12 -
252
-------
flj
a.
CO
«N
s
I
a
in
.a
I
x
O
UL
il
253
-------
Figure 2.
100
50-
50-
\.
8-
Treated Sewage
a— „ ,a
Treated Sewage
»8 Industrial Waste
.A Industrial Waste
'10 r15
DAYS
T20
25
254
-------
Rgure 3.
150-
<
o.
100-
\/\
V
V
70- *
»^ Raw Sewage
Treated Sewage
-2,
Control A
Industrial Waste
3—
Control 8
A
10 ' '20
DAYS
V
33
255
-------
180-
ngure
100-
Control B
a
J
O
100-
50-
0
-rt
lU
1 '20
DAYS
30
256
-------
figure
150-
1
Control A ,,
100-
o
z
50-.
ControJ B
10
DAYS
257
'20
'30
-------
MIDWEST RESEARCH INST1T1
425VolkerBoule
Kansas City, Missouri 6
Telephone (816) 753-;
February 1, 1978
Dr. W. A. Telyard, Chief
Energy and Mining Branch
Effluent Guidelines Division
WH-552
Environmental Protection Agency
401 M Street, S.W.
Washington, D.C. 20460
Dear Bill:
Enclosed is a drawing of the liquid-liquid extractor we used for extracting
tannery wastewaters for base/neutral and acidic priority pollutants. The
design was patterned after the Hershberg-Wolf extractor sold by Ace Glass.
The precision bore leveling device was eliminated to facilitate getting them
easily fabricated locally.
This simplified model is not totally automated. The stopcock requires periodic
adjustment to maintain the appropriate solvent level in the sample chamber.
As I told Gail, we have designed a simple modification of the solvent return
to eliminate periodic readjustments but have not had time to check it out to
our satisfaction. We'll pass along this modification as soon as we can.
Please let me know if we can help further.
Best regards,
CSa
Clarence L. Haile
Senior Chemist
Program Manager,
Mass Spec Center
Enclosure
CLH:1m
259
-------
2.5cm l.D.
250ml
5 24/4°
24/40 ^. t—i -1—
45/50
TFE Sfopcock
1 cm l.D
?fiO
-------
Attendees
Seminar on Analytical Methods
Environmental Protection Agency
November 9 & 10, 1977
Charles E. Stephan, Chemist
U.S. EPA
6201 Cangdon Blvd.
Duluth, Minnesota 55804
218-727-6692 X510/ FTS: 783-9510
Phil Cook
EPA Duluth
6201 Congdon Blvd.
Duluth, Mn 55804
Ian M. Stewart, Manager
Electron Optics Group
Walter C. McCrone Assoc. Inc.
2820 S. Michigan Ave.,
Chicago, Illinois 60616
(312) 342-7100
John D. Hallett, Staff Engineer
Shell Oil Co.
P.O. Box 2463
Houston, Texas 77068
(713) 241-5778
Gary Seidel, Chemist
Bunker Hill
1508 Northwest Blvd.
Coeur d'Alene, Idaho 83814
(208) 667-6797
Stephen Wright, Lab Manager
Edward C. Jordan, Co., Inc.
Portland, Maine
(207) 775-5401
Joe C. Watt
Environmental Development Coordinator
Catalytic Inc.
1500 Market St.,
Philadelphia, Pennsylvania 19101
(215) 864-8109
261
-------
Bruce W. Long
Associates
Ryckman, Edgerley, Tomlinson & Asso., Inc.
12161 Lackland Road
St. Louis, Mo. 63141
(314) 434-6960
Carol A. Hammer, Associate
RETA/Envirodyne Engineers
12161 Lackland Road,
St. Louis Mo. 63141
(314) 434-6960
Robert A. Fluegge, Program Manager
Carborundum
Niagara Falls, New York
(716) 278-2992
Bernard S. MacCabe
Business Development Manager
Carburundum Co,
P.O. Box 1054
Niagara Falls, New York 14302
(716) 278-6347
E. Ellen Gonter, Manager
Water Laboratories Department
Cyrus Wm. Rice Division, NUS Corp.
15 Noble Avenue,
Pittsburgh, Pennsylvania 15205
(412) 343-9200
Liz Privitera
Environmental Scientist
Calspan Corp.
4455 Geneva Street,
P.O. Box 235
Buffalo, New York 14221
(716) 632-7500
Barry Langer
Chemical Engineer
Burns and Roe
P.O. Box 663,
Paramus, New Jersey
(201) 265-8710
Dr. Joseph N. Blazevich, Chemist
EPA, Region X, Lab
1555 Alaskan Way So.
Seattle, Washington 98134
442-5840/ FTS 8-399-5840
-------
David C. Hemphill, Chemist
U.S. EPA
EMSL/Las Vegas
P.O. Box 15027
Las Vegas, Nevada 89114
(702) 736-29697 FTS: 595-2969
Walter Shackelford, Research Chemist
U.S. EPA - Athens ERL
Athens, Georgia
(404) 546-3186
E. William Loy, Jr., Chemist
U.S. EPA, S & A Division, Region X
College Station Rd.
Athens, Georgia 30605
FTS: 250-3165/ Commercial (404)546-3165
Edward Taylor, Chief
Chemistry Section
Region I EPA - New England Regional Lab
60 Westview
Lexington, Ma
(617) 861-6700
Larry A. Parker, Chief
Laboratory Section
U.S. EPA, Region III, Wheeling, WV
303 Methodist 81dg.,
Wheeling, West Virginia 26003
(304) 233-127V FTS: 923-1049
Walter E. Andrews, Chief
Rochester Program Support Branch
U.S. EPA, Region II
Rochester, New York
(716) 473-3166
Francis T. Brezenskis, Laboratory Director
EPA, Region II
Hilton Inn
Fred Haeberer, Research Chemist
EPA - Athens, Georgia
College Station Rd.,
Athens, Georgia 30605
(404) 546-3781
Bill Donaldson, Chief
Analytical Chemistry Branch
U.S. EPA (Athens Environmental Research Lab,
-------
Dr. Larry D. Johnson, Research Chemist
U.S. EPA, Industrial Environmental Research Lab., R.T.P,
Research Triangle Park,
NC 27711
FTS: 629-2557
Commercial: (919) 541-2557
Dr. T.O. Munson, Chief
Organics Analysis Unit
U.S. EPA Annapolis Field Office
Annapolis Science Center
Annapolis, Maryland 21401
(301) 224-2740/ FTS: 922-3753
Thomas Bellar, Research Chemist
EPA - EMSL
Cincinnati, Ohio 45226
(513) 684-7311
Kathleen A. Carl berg, Chemist
EPA - Nat1! Enforcement Investigations Center
Bldg. 53, Denver Federal Center,
Denver, Colorado 80225
(303) 234-4661
Bob Claeys, NCASI
Engineering Experiment Station, O.S.U.
Corvallis, Oregon 97331
(503) 754-2015
O.J. Loaspon II, Chemist
U.S. EPA, NEIC
Box 25227, Bldg. 53 DFC
Denver, Colorado 80225
(303) 234-4661
Billy Fair!ess, Deputy Director
EPA
1819 W. Pershing
Chicago, Illinois
(312) 353-8370
Mark J. Carter, Deputy Chief
Chemistry Branch
EPA - NEIC
P.O. Box 25227, Bldg. 53
Federal Center
Denver, Colorado 80225
(303) 234-4661
-------
Gerard F. McKenna
Reg. Q.A. Coordinator
EPA - Region II
Edison, New Jersey
FTS: 340-6645/ (201) 321-6645
Richard D. Spear, Chief
Surv. & Monitor Branch
EPA - Region II,
Edison, New Jersey
8-340-6685/6 - 321-6685/6
James J. lichtenberg, Chief
ORganic Analyses Section
U.S. EPA
EMSL - Ci
(513) 684-7308
P. Michael Terlecky, Head
Environmental Science Section
Calspan Corporation
P.O. Box 235
Buffalo, New York 14221
(716) 632-7500
Martha Bronstein, Chemist
Calspan Corporation
Box 235
Buf-alo, New York
(716) 632-7500
Larry Wapensky, Organic Chemist
U.S. EPA - Region VIII
Box 25366 DFC
Denver, Colorado 80225
(303) 985-7725
C. H. Anderson, Research Chemist
U.S. EPA
Athens, Georgia
(404) 546-3452
Leon Myers, Sup. Research Chemist
U.S. EPA, RSKERL
Box 1198,
Ada, Ok
(405) 332-8800 Ext. 202
William B. Prescott, Manager
Research Services
American Cyanamid Company
Bound Brook, New Jersey 08805
(201) 356-2000 X2167
-------
Richard A. Javick, Senior Res. Chemist
FMC Corporation
Box 8
Princeton, New Jersey 08540
(609) 452-2300 X328
Robert T. Rosen, Research Chemist
Mass Spectroscopist
FMC Corporation
P.O. Box 8
Princeton, New Jersey 08540
(609) 452-2300
Dr. S. T. Mayre, Staff Chemist
Duke Power Company
422 South Church St.
Charlotte, North Carolina 28242
(704) 373-8283
Dr. S. C. Blum, Research Associate
Exxon Research and Engineering Co.
P.O. Box 121,
Linden, New Jersey 07036
(201) 474-3303
Frank Hochgesang
Environmental Analytical Coordinator
Mobil Research S Development Corp.
Bill ingsport Rd.
Paulsboro, New Jersey 08066
(609) 423-1040 X2479
Robert F. Sabcock, Research Chemist
Standard Oil Co. (Ind.)
P.O. Box 400
Naperville, Illinois 60540
(312) 420-5229
R. 0. Kagel, Senior Research Specialist
Dow Chemical Co.
574 Bldg.
Midland, Michigan 48640
(517) 636-2953
R. M. Dille, Supervisor
Texaco Inc.
P.O. 1108
Port Arthur, Tx
(713) 982-5711
Dr. R. F. Stubbeman, Section Leader
Celanese Chemical
P.O. Box 9077
Corpus Christi, Texas 78408
(512) 241-2343
-------
P.A. Wadsworth, Staff Research Physicist
Shell Development Co.
P.O. Box 1380
Houston, Texas 77001
(713) 493-7723
James E. Norn's, Group Leader
Analytical Environmental Technology Dept.
CIBA - Geigy Corp.
P.O. Box 113,
Mclntosh, AL 36553
(205) 944-2201
Judith Thatcher, Sr. Environmental Assoc.
American Petroleum Institute
2101 L St. N.W.
(202) 457-7079
Max Lazar, Manager
Quality Control
Hoffman - LaRoche (Representing PMA)
P.O. Box 238
Belvidere, New Jersey
(201) 475-5381
Gary D. Raw!ings, Sr. Research Engineer
Monsanto Research Corp.
1515 Nicholas Rd.
Dayton, Ohio
(513) 268-3411
William G. Krochta, Sr. Supervisor Analytical
PPG Industries
Box 31
Barberton, OH 44203
753-4561
Will M. Oil 1 son, Staff Chemist
API (2101 L St., N.W.)
Washington, D.C. 20037
(202) 457-73757 333-7711
George Stanko, Sr. Research Chemist
Shell Development Co. (Also MCA & API Rep.)
Box 1380
Houston, Texas 77001
(713) 493-7702
Charles P. Hensley, Chemist
EPA Region VII
25 Funston Rd.
Kansas City, Kansas 66115
(816) 374-42857 FTS: 758-4285
-------
Will 1am F. lully, Project Scientist
U.C.C.
So. Charleston, West Virginia
(304) 747-4755
Bob Fisher, Research Chemist
National Council Paper Industry
3434
Gainesville, Florida 32608
John W. Way, Research Supervisor
E. I. Dupont DeNemours & Co.
Industrial Chemistry Oept.
Experimental Station
81dg. 336
Wilmington, Delaware 19895
(302) 772-4376
Roger 0. Holm, Group Leader
Waste Water Effluents
Monsanto Research
1515 Nicholas Rd.
(513) 268-3411 X354, 385
Paul X. Riccobono, Manager
Materials Evaluation
J.P. Stevens
Garfield, New Jersey
Janine Neils, Manager
Laboratory Service
MRI/Northside Div.
10701 Red Circle Drive
Minnetou
(612) 933-7880
Clarence L. Haile, Program Manager
for Mass Spec
Midwest Research Institute
425 Volke Blvd.
Kansas City, Mo 64110
(816) 753-7600
M.L. (Bud) Moberg, President
Analytical Research Labs Inc.
160 Taylor St.
Monrovia, California 91016
(213) 357-3247
Robert Z. Muggli, Sr. Research Chemist
W.C. McCrone Associates
2820 S. Michigan Avenue
Chicago, Illinois 60616
(312) 842-7100
-------
Roderick A. Carr, Sr. Project Manager
Versar, Inc.
6621 Electronic Drive
Springfield, Va. 22151
(703) 750-3000
Richard Kearns, Manager
Field Sampling Operations
Hamilton Standard
Airport Rd.
Windsor Locks, Ct. 06096
(203) 623-1621 ext. 8868
H. V. Myers, Consulting Engineer
NUS Corooration
Manor Oak Two,
1910 Cochran Rd.,
Pittsburgh, Pennsylvania 15220
(412) 343-9200
Jack R. Hall, Manager
Analytical Services
Hydroscience
9041 Executive Park Drive
Knoxville, Tenn. 37919
(615) 690-3211
Linda B. Kay
Environmental Scientist
Versar, Inc.
6621 Electronic Dr.
Springfield, Va.
(703) 750-3000
Jay L. Crane, Project Manager
Jacobs Engineering
251 So. Lake Ave.,
Pasadena, California 91101
(213) 449-2171
Bonnie Parrott, Environmental Engineer
Jacobs Engineering Co.
251 So. Lake
Pasadena, California 91101
(213) 449-2171
H. Dwight Fisher
V.P. Technical Director
West Coast technical Service, Inc.
17605 Fabrica Way
Cerritos, Ca. 90701
Jack Northington, Asst. Technical Director -> / &
West Coast Technical Director
17605 Fabrica Wav. Suit.p n
-------
10
Jim Spigarelli, Associate Director
for Analytical Chemistry
Midwest Research Institute
425 Volker Blvd.
Kansas City, Mo 64110
(816) 753-7600
Authur J. Condren, Manager
Analytical Services
E. C. Jordan Co.
PP.0. Box 7050,
Downtown Station,
Portland, Maine 04112
(207) 775-5401
Donald M. Shilesky, Project Manager
SCS Engineer
11800 Sunrise Valley Dr., Suite 432
(703) 620-3677
John H. Taylor, Laboratory Director
Jacobs Engineering Co.
660 S. Fair Oaks,
Pasadena, California 91105
(213) 795-7553
Charlie Westerman, Sr. Chemist
Environmental Science & Engineering, Inc.
P.O. Box 13454
Gainesville, Florida 32604
(904) 372-3318
Paul A. Taylor, President
California Analytical Lab., Inc.
401 N. 16th St.
Sacramento, California
(916) 444-9602
David D. Conway, Supervisor
Conservation Section
Marathon Oil Co.
P.O. Box 269
Littleton, Colorado
(303) 794-2601
Robert D. Kleopfer, Chief
EPA, Region VII
25 Funston Rd.,
Kansas City, Kansas 66115
(816) 374-428S/ FTS: 758-4285
-------
11
Charles W. Amelotti
Sverdrup & Parcel and Associates
800 N. 12th Blvd
St. Louis, Mo 63101
(314) 436-7600
Stuart A. Whitlock, Manager
Organic Chemistry Group
Environmental Science & Eng. Inc.
P.O. 13454
University Station
Gainesville, Florida
((904) 372-3318
Kendall B. Randolph, Chemical Engineer
Versar, Inc.
6621 Electronic Dr.,
Springfield, Va.
750-3000
D.R. Rushneck
PJ8 Labs
Pasadena, California 91101
Rick Johnston, AA Specialist
Edward H. Richardson Associates
P.O. Box 935
Dover, Delaware 19901
(302) 697-2183
Donald R. Wilkinson, Ph.D.
Director of Organic Analyses
Edward H. Richardson Associates
Dover, Delaware 19901
(302) 697-2183
Ronald G. Oldhan, Sr. Staff Scientist
Radian Corp.
P.O. Box 9948
Austin, Texas 78756
(512) 454-4797
Larry Keith, Head
Organic Chemistry Department
Radian Corporation
P.O. Box 9948
Austin, Texas 78766
(512) 45404797
James K. Rice, Consulting Engineer
Utility Water Act Group
17415 Batchellors Forest Rd.
Olney, Maryland 20832 ^ n O
(301) 774-2210 * '
-------
James Ryan, Manager
Gulf South Research Institute
P.O. Box 70186
(504) 283-4223
7
-------
r-
U.S. Environmenta! Protection Agency
Region V, Library
230 South Dearborn Street
Chicago. Illinois 60604
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