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

-------

-------
                   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

-------
                         Table  of Content
                                                     Page
I.    Proceedings                                      1
II.   Coimients                                        80
III.  References                                      156
IV.   Attendees                                       261

-------
                           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

-------
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

-------
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.

-------
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

-------
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

-------
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.

-------
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

-------
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

-------
    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:

-------
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?

-------
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
                               10

-------
n

-------
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
                             12

-------
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.
                            13

-------
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.
                              1 4

-------
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).
                             15

-------
    (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
                             16

-------
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
                              1 7

-------
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

-------
 o
   'jj

3 CE
LU 00 * •*
                       (A
                       LU
                       B
                       r>'
•o -a r>. — oo i
                                                                    I O O O i
                                                                         80 > (
                                                                         oo n <
                                                                          i  o  o  o •
                                                                          '  n  o  N <
                                                                          i  n  o  -o i
                                                                      OOOOOO

                                                                                >
I  O O <
>  m -o <
>  r-. in <
                                                                                                                     -— oooo — MO
                                                                                                                        —             IN
O O 4) O O O
•0 111 V -0 -0 Is-

~ 00 >*> N -0 <">
w.
LU
moo
zoo
LU
u. in h«
tjj-n

5
1
§O
8
(0 
^
«•


88
00

ct ^
n


i i
o o
o o
o o
51



So
0
o in
3S


0 0
So
0

^ 01






II
» C1)
•M 1*)


o
o
o

o




o
o
o
3



o
o
^



o
8

"•»






8
o
^
f*


1

o




o
o
o
s



o
„



8

»






o
o
o
T
^


o
o
o

**




o
o
o
R



o
")



o
o
o

l-o»«r(N
-^«N -L!:-?3^3?ii


OOOOOOOOOOOOOOOOOOOOO
ooooooooooooooooooooo
OOIOO»>coi>o.io-o-'B|V'Biv^»,^».a,oO»O.BO-'«'T'N
-* -^) I"1* 04 — -• «- iN il") •• '00 * ^ tf) 0^ — O


o
g

o




1
 > l>
— -* M



















j

cc
5
L

C
i
1^
5
1
z
m -

o in m - co oo
O 00 «r » '? ^
r'l >y rv * *- a^

^ '33 IV -0 '1J f*
(.-1 -. pv ,1) If, l>
-0  '-1 (V •'•">
>*• -• -4 IN '-1 -0
i) ri
in
1
ri 5 $ '•<> li j"j
? ;P >)3 ci .» O
— '.0 CO «• '7- iv
1


-, .; in » » c

J2 oo — ^ S *
'B ' » — — i-l
l'l ^

i


c - 'n -o —  5 -T
-u ^ O -0 '> -0
X) * v T «r ^
;. .> i'O r| :/J ^
                                                                                                                                                             j-> - -.
                                                                                                                                                       m    in
                                                                    I o <
                                                                    : 2 ;
                                                                    i o <
                                                       OOOOUGO(J(J(J(JO(OQ)U)(AV)U1'J)i/lU)U>iJ)U]a)U>fa)
                                                                                                                                                    S i g 8 i
                                                                                                                                                    «
-------
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

-------

^ "O
INE CREEK-UCC
IOLYCORP-QUESTA




0



H H
> 33
— m
j^
Z H
EDMINE WATE
GPONDEFFLU
m 33
Z
H










CO CO
CO CO
x x
o o
-» -si
o


N) CO
O N>
X X

c. c
O 
i ~™
r~ 1.
§ Z
> O
H TJ
m Q
33 Z
H O
Ojj
33 m
m O
0 •<
^ o
o r-
r~ rn
m










-^ sj
en sj
x x
o o
CO CO



-» en
CO -sj
X X

o o



X O
OTTER-SCHWARTZV
ERR-McGEE
«-~
r~
O
m
35
C C



H H
33 33
m m

H H
m m
0 0
S 2
Z Z
m m
§ $
> >
H H
m m
33 33











A N>
co co
X X
o o
CO CO



(71 NJ
CO O
X X

o o
sj CO


r c 35
EPUBLIC
'CC-URAVAN
UCKY Me MINING




C C J1



H m H
33 Tl >
m TI —
t" r™ '
H C Z
mmo
i^i
?S§o
§|?
> 2 -n
H - r
m £ c
35 £ m
— I ~*
H
r~
Z
(7)
13
0
Z
D


en -a j^
sj NJ CO
xxx
o o o
co co s.



NJ _> JS.
•sj cn — »
xxx

o o o
•st co cr


T CO
T.JOE-EDWARDS
ANNA-BUVLER




-n -o
(B O"
N
2

2 H
Z -
m r;
•s z
G POND EFFLU
/ATERSETTLIN
2|

z
o








-t^ CO
N> J^
X X
o o
sj CO



CO NJ
CO £>
X X

o o
OT sj


Z CO
UNKER HILL
ECLA-STAR




-0 13
cr cr
N N
w 3

H H
> 33
— m
— >
Z H
MENT SYSTEM
G POND EFFLU
m rn
H ^H
C
m
Z
H






— > -P>
0) ->
X X
0 0
U3 CO


A
CO •&
CO ->
X X

0 O
en -j


> >
NACONDA-BUTTE
.NACONDA-BUTTE




o o
c c



H H
33 >
m —
j>
H Z
G PONDEFFLU
ED MINE WATE
33 m
Z
H










sj — »
N) N)
X X
0 0




CO CO
NJ O
X X

C 0
O) O


$ X
ENNECOTT-SLC
fHITE PINE




o n
c c
•g
CO
H
H H
33 33
m m
^ ^
H H
MENT PLANT E
MENT SYSTEM
rn -n
Tl Tl
TI r~
r- c
C m
m 2
Z H
H






CO £>
NJ CO
X X
o o
en co



en sj
en ^j
x x

o o
3 on «4


XS /N
ENNECOTT-SLC
ENNECOTT-SLC




0 O
c c.
3 3

£. Z
H H
> 33
— m
n >
Z H
MENT PLANT E
G POND EFFLU
m TI
H £
m
Z
H







CO -»
>j en
x x
0 0
sj «sl



CO >J
N) CO
X X

o o
en en


X >
SARCO-GALENA
ENNECOTT-SLC




p >
C 4Q
3
NJ
CO
H H
~~ ~~
r~ r~
Z Z
GPOND EFFLU
G POND EFFLU
m rn
Z Z
H H










*• NJ
CO ->
X X
O 0
to co



OT -»
sj CO
X X

o o
CO CO


> >
LCOA
SARCO-GALENA




>>



H H
33 33
m m
^ ^
H H
EDMINE WATE
EDMINE WATE
33 33











en -»
•si *>
x x
O 0




-i M
-» O
x x

0 0
o co



FACILITY




O
33
m



_
^
ASTEWATER SC
C
33
O
m







H
— O
""** TJ
i1 >
5. r
~ —
 CO
C, m
33
n
~ X
cr 33

3 CO
=: O
f+ ~~l
O — •
— r
m
                                             >
                                             CO
                                             r~
                                             m
                                             33
                                             m
                                             co
                                             C
                                             co

                                             O
                                             Tl
                                             CO
                                             O
                                             33
                                             m
                                             m
                                             Z
                                             co
                                             TJ
                                             r-
                                             m
                                             CO

                                             CO

                                             o
                                             Tl

                                             H
                                             Tl

                                             5
                                             m
                                             33
                                             o
                                             o
                                             X
                                             33



                                             I
                                             F
                                             m
                                             >
                                             co
                                             CO
                                             m
                                             O
                                             co
46

-------
O

!/5
UJ
CO
LU
_1


O
oo

CC
X
o
Q
Z


cc
UJ
CO
    UJ
O
I-  <
UL  >
O  LU
CO
    uj
     I
-J  UJ
<  UJ
2  H
<  O)

uj  oo
_j  -;
Q.  2.
CO

2
ai
UJ
cc
u.
O
 CO
 UJ
 CC
 UJ
 _J
 ca
 <
LU 	
— 03
t— . -
O^
co Z
tt|

cc^
00 G3
iZ ^
1 
 ™*
i

H
Ij
O
U.



^
o
X
U)
CO
CO
o
*~
X
00
CO*

•z.

\-
UJ
ORAGI
H
co
CC
UJ
t—
STEWA
<
g
CO
{M
O


UJ
z
i
<
>
UJ
Z

0
















,


o
u.
CC
UJ
>
o




jj
D
CARBON,
•
— •

CO
o
r"
X

LU
z
LU
U


















2
UJ
NEWAT
5
a.




^2
3
CARBON,
•
— •

ps.
O
r—
X
S
                                                      ^  X
                                                      2  PO
                                                      <  ro
 O
 2  S
 5  I]


 II
 CO  |_
 uj  G
 CC  UJ
 CC  I-
 O  uj
 O  Q
                                47

-------
CO
O
O
cc
at
CO
u.
LU
Z
g
z
a
2
2
D
O
IGHESTCHRYSOTILEC
(FIBERS/LITER)
Z


IOLYCORP- QUESTA
^


ENNECOTT-Cu-02B
^
ING EFFLUENT
_j
NACONDA-BUTTE - TAl
<
CC
LU
Q
O TTER-SCHWARTZWAL
LCOA
O <
G EFFLUENT
2
SARCO-GALENA - TAILI
<


ICC-URAVAN
^


ENNECOTT-CU-OSB
m


LACER-AMEX
a.


LU
UJ
O
0
cc
cc
UJ
^
cn
o
o
CM
CO
o
ft
co
CO
o
o
00
CO
o
6
CM
CO
o
e
c\i
CO
o
CO
r-
co
o
Lfl
r-
O
*
^Si
^^
o
fi.
in
o
V™
CO
LT)
z
X
LU

00
LU
U.

LU
cc
o
LU
_J
CQ

<
I-
1-
2
LU
D
_l
u.
1 1
UJ
COUNT
)
cc cc
at uj
CO H-
U. _J
HEST TOTAL
(FIBERS,
a
z
TAILING
'
<
h-
00
UJ
a
a.
cc
O
u
o
5
EFFLUENT
1-
REATMEIS
l_

CQ
CO
o
NECOTT-Cu-i
2
LU
^.
LUENT
u.
u.
LU
o
2
-J
<
h-

CQ
CN
O
NECOTT-Cu-
2
UJ
^t£
Q
UJ
<
LU
££
H
CC
LU
Q
_1
<

N
J-
re
TER-SCHWAI
E WATER
o|
o ^
EFFLUENT
Q
2
O
a.
O
2
13

<
*-

-------
     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
KENNECOTT-SLC
KENNECOTT-SLC
WHITE PINE
ANACONDA-BUTTE
ANACONDA-BUTTE
BUNKER HILL
H EC LA-STAR
ST. JOE-EDWARDS
HANNA-BUTLER
REPUBLIC
UCC-URAVAN
LUCKY McMINING
COTTER-SCHWARTZWALDER
KERR-McGEE
PLACE R-AIWEX
MclNTYRE DEVELOPMENT
PINE CREEK-UCC
MOLYCORP-QUESTA
ORE
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
U
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
0.39
64
240
0.28
2.9
27
0.19
110
2.9
14
<0.12
8.5
1.3
1.5
53
9.5
70
19
20
0.46
2.9
700
"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

-------
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

-------
o
<
o
o;
o_
D_
o
o
ce
Q.
                        <
                        D_
                               CO
                               •«—'
                               c
                               CT3
                               J_
                               O
<  53
Sit*
        O CO
                                              53

-------
—    S
03    -J
£§|
,-*-, i^i X
^ uj N—
Z C_) Qi
                   I
                            £i^
                            s
                   £   >-   i
                   K-   —   VI
                   z   o:
                            5
                                     S  2
                                     —
                                                            s


X z
<^» a.

>• £
0? iS
< <
§£
O r-
C_ ) wl
UJ <
^ g
_

S z
< 5
o=

                                                                                   J:   5
                                                                                   —   a_

                                                                                   o

                                                                                   a;
                                                                                   o-   )±i

                       o   5
                       S   OS
                       Q.   ji;

                                        54

-------
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
                           55

-------
                                        CD  ^_

                                       .E  £

                                        fe  c
CD

-------
_1    <5

S > 8
CK O O
I— O a:
ce o  s
a: uj  ^

tl     ri
       O
1  -  >•

3  U  Q


t  ><  2
U.  0  —

UJ  >—

o  uj  5/2
en
UJ
I—
<
o o  a.
UJ "J  <

5
a-     <
   >-
   I—
   o


   e
   o
                                   cc
                                   o
                          t^1 o >—
                          I— ^ H~
                          <: 5 o
                          SS33:
ii
£ <
       s

-------
 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

-------
<     o
— >•!_>
Q£ O  O

£ 2  g

£ 8  5
CC LU  *=
UJ     _J

       O
       on
    t  >
    cj
~
uj
I—
<
   S  £
O o  Q.

LU UJ  <
   >-
   )—


   O
   O

   >-
                                  a.
                                  S
                                  o;
   X    x_

   °    £
t>r  ^ ^J  c_J

"  ° i="  X
<  § o  o
g  S. CO  ^~
uZ  S uu  <
on  _ <  5

~  < °  ?

^  = <  <

   <    °
O X
< Q-
l— >-
Z3 I—
S <
if
ce  5
    63

-------
64

-------
£  ^   8
cc  o   o
•—  O   a:
""!  si   <->
>-*J  O   —•
o:  o   s
cr  ua   •=
         o
         0-1
=   o
<
o:
o:
LU
K-

<
s  °
O  o
         <


         
S   1/1     <
     O-     ce

     a     o


     L/l
                                       S     S
                                  i   3 S  S
                                  t;   o i—  x
                                  <   I P  2
                                  QJ   Q 2  z
                                  ce   < _j  ?
>:  B
n  °£
      65

-------
Teflon Lined
PVC Tube
  Soil Depth
  0-5 cm.

 Teflon Ring
                                                            Inverted Beaker
                                                            Alkali Trap
                                                            Glass Funnel
                                                            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

A
B
C

D
E
F

G




code c«st

S N.P.*
SE N.P.
V » P.

DD N.P
CC "J.p
L N.P.

F N P



Cytotoxieity T.iti. pjthead
% erf. cone. * eff. cone. (LC^Q)

N.D. EC20 " 0.58 19.0
N.D. N.D. N.A.T.C
EC;0 . 9.P° FC50 " 3.35* 46.5
Ew _ u a l.bb L^JO J.o<
N.D. N.D. N. A T.
N.D. N D. N.A.T.
S D. EC5a * 35.0 M.A.T.
EC2C • 0.94
N'.D. EC20 - 17.3 64.7
g



(LC50I

9.0
N.A.T.
41.0

N A.T.
1.8
81.7

62.4

1001 cone

Sheepshead
(LC53) (LC;0)

62.0
5 G >1CO
H.G. 69.5

S.G -f
N . G . > LOQ
H G. >100

H.G. >100
f


ne Ecoloa/ Tests
(LCsol (ECr,o) range K ' m 'Vq

21.2 - >1C
>100 -3 20 >1G
12.8 90 1.3 MO

-f -f -f MO
>1QO 10 to 50 -f >1C
MOO 35 2 3 MO

MOO 59 1.3 MO
t f f _1C

N.P

N.P

N.P
N.D.

N.D.

M.D.
EC;c * 1-H   N.A.T.

   N.D.       N.A.T.
                               3  52     23 5
                               0. 40
N.A.T.

N.A.T.

 28.3
S.G.

S.G
         '100

         MOO
                                                           MOO

                                                           MOO
                                                70

-------
LU


h—
OO
LU
I—
O

CQ
<
X

CU
ry wastewal
fD
•o
c
o
CD
oo
oo
CD
"o
CQ



ro



CNJ




-
-

0
u_

LU


Q



O

CQ
-
"c -g
o_ <->
LA
CVJ
I— 1
0
CNJ

1
1
1


CD


CD
S
LA
i— i
CD


O
0
c — 1
A
o
o
, — 1
A

0


CD



0

CD
CD
CQ

CD
CD
^
LA
ro

o£


ro


C\J
LA
LA

"
CD
o

CNJ
ro
sO
CXI
^p

c
oo
T3
^^3 ,«•••
f\ "\ ^^^^ «^
ro ^;
o =5
r— 1

oo
oo
ro
I— 1
ro
Q

O
CD
LA
ro
CD
OO
sO

C
o .2
oo J2
06 5
"on

0


LA
\^^
•>^J^
CD
CD
LA
CD
LU

                                      71

-------
      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

-------
O
— I
o
o
LU
CQ
               CD
2.
 a
 C
 o
.a
 i_
 ro
O

-O
 CD

"TO
                                              OJ
00
LU

CQ
              ra

              '•o
              CD
              E
 ro
 C
 ra
 i_

O
                        ra
                            ra

                            TD
                            CD

                            E
on
LU
o
O
              a>
                                            03

                                           "ro
                                            O
LU

              o
              ra
              CD
 ra

'•5
 CD

 E
 ca

'•a
 CD
 E
         ro
         C
         O
         M

        O
 ro

O
                        ro
                        CD
            ro

            ID

            S1
            O
            O
                                                                 -o
                                                                  CD
 (/•>



O
CE:
<
CD
Tl^ji
o
CO
CD
"^3
O
o
CD
•o
O
o
CD
•a
o
LU
CD
•a
o
* W
O
•^^
Q_
^— \
u_
CD
"O
o
o
CD
T3
O
                                   77

-------
i/l
z
o
Q-

5
<
                             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

-------



•





















Jy.
Q)
£>
l-i
3
Cfl

07
0)
u
c
c
4J
to
rC!
3
CO
r\j
I N
O
4) -r-l
rH X
rC O
<3 E-
E-1
to
O
C
•H
U-l

S
y
O

rH
•H
0
-X CT
Cj
U-J C.
0 M

C.
o
07
• r-l
U
d
a
E
O
U




















< tn
c- :•-
u y
w




,H
• H CT.
,a 2

A


r-l
°1"4 eJ
*§ y
2 0




rQ ^
o y
2 O
























cnpnc (-IP CD
22222 :^ 2
O


/~0^-^ r^^-s

rH
P P p •* CN in •t
0222 • •
O O O rH

/-^_rV •— S>C»— -V.
'^ ^ *S^~- -^

f^ CN CN 'J' rH n
O rH O O O O
\' V V V




C C Q P CO P in
2222- 2
rH O ^
rH



^- — J**~** ^~-^ — *~\


P C P P ^ P CT«
2222 • 2
rf CN vo
rH

'~~~_A-— >,
'"^-A— ^
CM
(N rsj* — im^^O rH V£) ^f
. . • .
ro o in 0^ vo ^^ in v^
^j* vo r^
V ^

/-~»^»— —
' "^ ~\ x-^-rs^-Vj

in CN 10
I . 1 .
rH ro^rroTj-cnino
in rH vO ro n
rH




Cl)
c
flj
u
Q) (3
C l-i
O CD CD 0) j^
0) rH C C C 4J
C >i O CD Q) C C
O rC X: rJ C rC <
r*H i j i tt Q) p" (ii i QJ ^_^^
fl j;: .c c. .iJ u c c c
rCCX.iiOcraiTjoaj^
4-lTOTJr-ir31-lr^Ct/lO
rCcco^^rocj>,N
D-,O(l;3^4J3>-il-iC
a u (jrHrC: c--i >,jr a)
Zv_/<^__^

TJ- VD
O O
V V




g




/— ^\— -N



CO
•
rH


r*^^JL.


n rH
o o

V





p
2


/---'v^-v
^+.
.
VD


<-^\_— ,


in ^H
• .
•H O
V


0 0)
c c
O 0)
rC ,C
4-1 4J
c c
fO (3
1-1 r-l
O 0
D 3
rH rH
U, t.
-0 ^
• 	 . . 	 .
0 0
N N
C C
Q 0)
CQ «
fl
^-^
ro
•
rH


— .
ro
^ — •
CM
•
r-i




•^
O
V

~~-
ra
*— '
in
CT>






,— .
(3
^^
sD
rH


s.

cn
H





(C

ro
fl


X
2
*— '
VO
rH

t



1










0)
c
,
CL
re
- — •
o
N
c
0)
CQ


p
2






p
2





VD
O
V




1





,


o
•
rH





r-l
o

V










CO
.
o





CN
•
^

fl)
c
D
r-l
£x,
a.
.r-^
T3
u
1
n
.
CN
%
x_-
0
c
o
•D
c
M


p
2






p
2





U3
o
V




Q
2








P
2






•-1
O







Q
2




P
2





in

o

o
c
0)
u
A3
)-i
,c
4-1
C
<

jr;
%
i3
O
N
C
O

.rH
P


a
2






Q
2





r~
O
V




6
*-*








^
•
CN





in
rH







Q
J3





in





CO

m



0)
c
0)
rH
>!
r-l
CU
CU
^_.
•r-l
.
f.
CT1
• —
0

c
V
o




•
Q)
3
i — t
13
£»

tj
CJ
C
Vj
E
O
U


• H
U)
•H
^c
4J
,^

CJ
4J
•H
U)

0)
w
<1J
jf^
4J

SH
0
U-l



U)
•H
n
CT1
C
4J
cn
• rH
t!

(D
rQ
_jj
O
C
c
CJ

a)
c
0)
r-l
U

Cu

•o
c
rd

Cu
(C
ffl

^^
(3
• — •













)-i
(3
rH
3
U
O
.H
o


0)
c
o
rH
>1
to
C
a
vw
O

-Q
Cu
a

CO
(•*•>

4-1
U
o
4-1
0)
T3

O
aJ

a
rH
J3
rl
1 J
U7
(3
3J

rH
Vj
O
2

-
O
4J
>r^
V)

en
C
•H
i— I
a, •
§1—1
rH
in o
5
CQ t/7
Cu <3
M
- — •
O rsi
X! in
4-> CN

to.
O
b.

'^
^
^— ^








93

-------
                         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

-------
                         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

-------
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

-------
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

-------
                 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

-------
                                                          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

-------
                                                                  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

-------
                  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

-------
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

-------
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

-------
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

-------
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-

-------
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
.„
^
Q.S2
Q.9C
(0.9 s-;
a...
a.n
J.OG
/.oo
,.o
,.-
O.SS
Q.fO
|.3V
^
=55-
1 3?
!tt-
I US
U3
(is)
no.
((.7
*?lj ^
•5 ^4 S
15:0
ns
Wl
i ^ y
l^i Q
m
?2^
1.11 ' ?°^
--
lol
,c
S7
w
^
9,
/QO
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
0,,
,.M.
,.«
0.^
,.^








;
—
?1?
\ft
IM1
1 ^J (
1*7^
!,!r
OOJ-O
pocr-°°





\


i
J^,
^^
i,
,3
„
-








i





—








-- "".- ;'























' 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

=H
^
/O/
jaj
(jl
SI.
">«
01
i'_«.a»sS8SS


















H
;
|i

1




S^^^'^:
u- 	 ' " M" '135 -• '- 	 "-;• /r??^55T^^«^«
                            '- --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

0-SI

p 9 n


"7 1

o.st

1.0-1

1-14

i ii"?
'


wr.
•
W

~^
.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



•



(S.I3.T
0.73
\.\^
\,11.
1.34
o.a^
1.13






i

-
IS?.
^2 'i
—
-
-




I

1

95-
97
I3C
=?/
9*
H*
HI
«?«
/ >4<*
M"?
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.
*.«
*n
C.fl
l.oo
o.rz
1.37
,.-»
2,'t
1.1*
,.«
»«
,^
l.is"/- Qf"*
<3JdO~
-TO
!«
..»
^.i?
0,73
l.oo
o.t*
,.*
/.»*
«<
w/
J. 3?
,,,
,,,.
1











L





















•" "






















I i















Endosulfan  suU'ate     •   34351
                          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

-------
50
CD
rt,
H
rr
CD

n
o

CL
(D
3
w
CD
n

CD

H-
O
P
a
H-
rt
rt
H-
O

P
 tn
 H-
 m
 CL
 H-
 rt
 H-
 O
 3
 P
 O
 M
 rt
 H
 P

 CL


 73
 CD
 H-
 3

 O
 O

-I
 P

X

S en
x o
C. CL
rt H-
O C

H«
CL
CD






S f n

/-> H- CD C
2 rt in
P CD rt
O H-
so rt
^^f p
C CO
W O
rt CL
H- p
0


n
o

Hj
o
yj
H-
^
(T>


-o
o
t-" O
1—" rt
CD X
rt
M


/~~\ t~*
w o
o
o
N OQ
• 3
v_- .
p !/i rt rt) S
3 o rt H- zr
CL i— ' X P H-
C tn ?T* rt
TO Cf rt CD O
> — ' i — * P 'J) <*
X C5 t-> -
rt >-* CL

i-i 3 3 C ^
O CD 3 H-
i-" s: "o .0
P rtj V) C
rt i-i CD
CD P O t/i
rt rt rt rt
• rt CD
C w 3
P H rt rt
(-• CD H-
rt » rt
O if
S* W
o
I—"

w 3 cr
XJ • .
w »
w
•


t-

N> i— ' to
• oo o
H- O O
woo


/-•> "O
n 3-
o o
CO 3 > cn
tn rt rt "3
e\° CD H- •y
\__j fc^ £^ i-\
rt H
H M-
P rt
rt
CD
CL


0

/— , rt
' ' 3"1
OJ > O
*3 rt *O
O H- rr
•*>• CL O
N — ' £/l
*^3
3"
O

H*
o

n
o
H

O
(/>
H-

CD


r-
H-
ff)
C
H-
CL



/~> i—1
^/J O
o
o
N g
• t~*
N 	 '
rt H- o >— n
i-i H- rt O i-1
"3 tn CD
rt O rt W p
-J. !-)„>•(
O C- P
3 fD 3 tn
*T3 tn *"O ^
p o ^ P o
3 3 P >•! — '
CL CL M 7T O
— • CD K- ' l-j
rt 3 3 "-• H^
O TO rt 3 O
3 OO tn
^3 O rt tn
CO 3 i-i i-1"
rt X "• 0
ft3 O 01 rQ C-t
rt O rt C 0
C 3 P H- rt
rt n 1— CL 1
CD CD 1
3
i

w 3
13 •
jq .

•H !
;
j

ts)
t— 1 •
* C-J '
CD VI
OJ C
** n ,

^-s en
n c
O >— '
3 rt>
 rt
rt rt
H- H-
CL O


n
o
rt

o

H'

CO


r4
H-
f-i
C
H-
CL



/ — > i—"
W o
0
0
N 3
• t— •
^
rt CL s cr n CT
P w. p HJ O O
C V) rt O I-1 3
tn './> O £ O cn
O O rt 3 rt CD
tn i— ' - * ?— -
< O
o n> < = t/i c
3" t/) O M. V) H-
p rt V) — '
rt 3 X n rt X
rt 0 -• 0
r-. yj rt cr i—
3 rt 0 r— CL -"•
Oq a CD P ^3
3 rt rt C
CD rt S 7T H-
rt H- H- CL
P < rt -
(-• CD 3*
t/) •
^




w 3 cr

fjQ . .
^
• ^i
O*i P-
I— 3
W 03
1— 1 CD
O W
)•— » • C^J
• £* CO
00 O O
^. n n

, — , 2
n H-
O rt
£-> 4
^j rt H-
o o n
o\« 3
^ rt >
rt rt
pj ui>
rt CL
CD
CL
i

•y m
/— \ N 3
" O O-Q
2, rt rt
OH- P
w rt <
v — f CD
2> rt
o
H- cn
CL

rt
CL

n o
O X

rt CL
O H-
W N
H- CD
^ rt
CD


n
H-
<->
£
H-
CL



/—> I-1
O-J O
O
o
N 3
» H-1
"— '
M tn O H
3 c rt rt
CL rt) p
rtiX 3
rt O CD W
o rt i— "a
rt P i—* P
i-i rt O rt
O H- S CD
00 3 1— 3
M- OQ t/1 rt
< - 3"
CD rt
O rtj O
>— P C (-1
H- C 3 0
X> VJ H- rt
C rt 3 >-
H- "-OQ CD
CL rt » tn
V ^J





< rt < 01 a cr
H- CD p T3 • •
• rt • • oo •
• (-">o rt
3 rt •
• •

O I
• ^_J ^
j •vJ • 1— " t— '
i cr* o*i c^ • * '~i
i— ' 'O fO CiT C^ ^
rt ^ ^ O O O
*^j o 2 -o. n n

n
3"
CD
3
H-
n
P
i— u

2
P
3
O

/ — v
n
3"
CD n
3 0
H- 3
n 3
P 0
t- 3

-n 2
O P
rt 3
3 CD
C
P
V — /
n
p

w E
H. p
rt) N
H. p
rt rt
p CL
(-f
H-
O
3

•n
o
rt
~3

1
t
XD
C
P
3
rt
H'
rt
X
|


,

. "3
rt
o
• "3
i CD
rt
rt
, H*
1 CD
' tn



!






S
: n
' P
f rt 2 '
• P p
rt rt ,
rt CD ;
CO rt '
' WJ ' 	 '
j H- Pj ,

rt
'- •
rt
ir.
m


wi
                                                                                                         n
n

c-

50
tn

CD
m
2
H

J^
t-H
H

n
o
c
m
                                           150

-------
A
   \


              /  \
                                                     \
                                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

-------
                             : rppr~U ; ~-Ti:rr; i 'i o i ^n;i:^T;i_;'::
                             -!-K-^!:^-:  -i  -r- !- -Kl  . rihll'l-f!:-!-
                                                      . -
                                                      i i .'  • :  i. ^._  !	
                                                      ~    """
                                       Lj_!J_..l_LU_LJ_Li_U_4_j_L .
                                       L.L J_i ._!_£' L l_iLtl. Xl ;


2    4   6    8    10   12
                                                        26   2^  50

-------
4-
                                         •4-!.; I.14-1 -4	-LJJ-J...  ,-Uf-J I  '.!. i i
                                         Li I...  J L 1..J 1 i_i JILL. .'-.I . _.j 1 L  1 . '.
                                                                                     ^CT^.oLz-% £
                                                                                      I  I —	p^ 'n '•
                                                                                     .'Ul- LT";  ' i

                                                               --.--J-H-hr


                                                                          1  t-  ' vi_ji  _.'  L;.

                                                                          ;  l-LlgWlH-'r-
                                                        JO     12    14    16    IB    20    12-    Z.A    26   28   30

-------
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

-------
                      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

-------
            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

-------






















4_J
C.
c.

rH




CO
01
rH
•H
E

0
O
O

0
0
o
M
\D
rH
-*x^
f.
CJ
C.
•H

rH






<0
^— ^
C C
O 3

O.
CO 0)
Ol jT
_ 1 j_j

JC 0
o ^-*
c

0)
X d
•H }-<
CO 3
o

^-*





CO
0)
•H
l-l
D
4J
a
a
. CJ

o
CM
CO

•o
c
o
CJ
o>
CO

rH









O
0
o
•%
o
o
0
A
O
o
o

o
I-l

-*^
•0
rH



03

6


o
o
o

o





t-J
O
t — f ;
C C ^

C
•— t
- cc
J
J U-.

U-l
rH
O
CC





i. - ~ O O


C cC
•r-i r3 —
CM CO
u~i CO 0-*
tC O CO 1^ rH




C- ^ -s
CO '^ --'


r c a -H
-H CJ g
3
-I .C 4=i cr
o
o
0
*
o
o
o







rH -u di 4-i C 0) 4-1 G 03
--x, 3 ^ " -H -H 3 CO
\ 1
rH
a
OJ

4=
0
C
•H
cx

rH
CO
(3.
-H
j^
U
O to C !-i u
E fcC s cu
l-l O l-l > r-
CJ 4= O C r-
^ > CJ
-^ 0 *-> o
e "^
-1 l-l rH CM
-i cu a ^-
"3 ^> CO C3
o
o
U"}
*«
CO
"**^


0)
4-1
C
01
C 43 -rl >, 2
O
4-1
cd
I 1
o

CuO C. S •«-> O-O
0 O 0 -H <

-a o -a u
u*i V-i u-J
rH CM O i— t C C
3 O O
3 )-i O cr
-1 T3 - CO
r-l O
3 O rH CO rH
OJ
6C
0
43

rH
a
c
)-!
3
O
4-1

CO
•H
C
c
01
4-1

0
0
0
"
o
o
0
*
o
o
CM
*v
i— 1 CO
"^^ CU
43 4H
O U
rH 4J
C3
r~H c


0
o
o
«
0
o
o
*«
o
0
o
•V
CM.
-^s.
cu
rH
cu
c.
CO CO
rH
•u cu
a \-i
43 l-i
c»3
r-l 43

o
c
o

o
o
o

o
iH
^ —
^4
0)
•u
c
a
UH

T3
a
4-1
C
01
T3

rH














CO
OJ
E
•H
4-1
OJ
ti-4
•H
rH

l-l
CJ
CJ
        to
        H
                             to
                             01
                            o
                            o
                            o
CJ

2
O
t— i
H
<
pi
2
0
U
f.
43
a.
? c.
o
rH rH
CJ
•H

rH




        w
                             CO
                             0)
                             e
                            vO
                             U
                             C
                            -H
                                             CO
                                             0)
                                            CO
                                             C
                                             O
                                             cj
                                             cu
                                             CO


O
o
o

0
o
o
M
o
rH
s,
Ol
to
o
43

rH











CO
4-1
C
o
£
C3
C
\^
3
O
4-1
s matches
1-1
c
c
0)
4-1

0
o
0
"
o
o
CM
«h
rH
"-X,
43
o
rH

rH
barrels
o
o
o
•t
0
0
0

CM
"•x^.
QJ
rH
O.
a
cj

13
03
43

rH
O
o
O
0

O
rH
^•~x
J-j
0)
•o
c
01
U-l

-a
0)
4-1
C
01
•a

rH













CO
O)
E
•H
4-1
0)
WH
•H
rH
 CO
 0)
 r-l
 O
 CO

CO
C^4
                                                                           cr
                                                                           CO
                                                                            O
                                                                            CO
                                                                                   C
                                                                                   O)
                                                                                   E
                                                                                   cd
                                                                                   c
                                                                                   l-l
                                                                                   3
                                                                                   O
                                                                                                                                O
                                                                                                                                60
                                                                                                                               O
                                                                                                                               O
         0)
         60
         O
        43
                                                                                                                                            CO
                                                                                                                                            O)
 CO
 s

 05
•H
 C

 CU
                                                                   O
                                                                   o
                                                                   CM
                                                                   43
                                                                   O
 crj
43

O
O
o
  •s
CM

 01
•H
 Cu

 CO

-a
 n)
43
                                                                                                                                                                    CO
                                                                                                                                                                    Ol
                                                                                                                                                                    E
                                                                                                                                                                   •H
                                                                                                                                                                    4-1
                                                                                                                                                                    0)
                                                                                                                                                                   U-l
                                                                                                                                                                    l-l
                                                                                                                                                                    CO
                                                                                                                                                                    O
                         01
                        •o
                         c
                         4)
                        U-4
                         C
                         ai
                        "3
                                                                                   176
                                                                           <

                                                                           §
                                                                                                                               u
                                                                                                                               <
                                                                                            _)
                                                                                            ^
                                                                                            o-
                                                                                                                                                                   fcj

-------
                     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

-------
                               FIGURE 4
                           r>
0
         A
         fl
                 c
1
I!
Ii
•5 J
* j
H
h
ii
n
V
f
;
j
i
i
L
el
|;
H.
3
5 '

1 •
1 ill
4 $ i
a
                           i!
i
si
                           i
                          « i


                          )
                        D,
                        n 3  3
                        I . "!


                        fll


                        •



                        1
       r5 ^?*r^rn'
       ^iJjUJil;
    D
    r>.
                              jl
           D

          f
  6
                                      8
 fi

 5
IA

11
" 3
  H
  :A
                                             'V«.
                                                       *>,

                                                       0 a
f\


'• ~\
                               v<
                                                        l.i
                             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

-------
                     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

-------
                      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

-------
26. Carborundum Corporation
27. TRW
28. Industrial Environmental Research Lab, Cincinnati
                             196

-------
                    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

-------
                   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)
                                    200

-------
      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
                             202

-------
           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

-------
     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
                            204

-------
           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.
                            205

-------
      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

-------
      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

-------
         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

-------
                                           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

-------
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

-------
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

-------
           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

-------
           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

-------
      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
                        214

-------
      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

-------
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

-------
              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

-------
       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
 t-.-i •—>••'• IP'.ii.'J b ^rr-^.1 ~^*p —?—#~ ******—jEV-T
x^a^ifi^asgsa^
-•\sr?*?52i:r?^^'.f.->7SSa3sH£
     "** i ^** - i?*^ T*V /- g:jt ij^^S^*.!^.^^_?^™**'-ji? -i\^^*^
                                        -^^fe^^S^S^?f^?E5^
                                        ^^^r^^^-rsggg^a^^
                                        ~.i^±-fe^:-?^' J> <-tP>"-^-|^fc5X^^^S:
 Probe of chunky material
Probe of large chialike material
           waiter c. me crone associates, inc.
                           235

-------
                         Sample
Probe of chrysotile
Probe of typical spherical particle
Probe of chrysotile bundle
                                   ^^^^^^^al^^^^S^&S^S^^S^sS^S
Probe of antigorite lath

                 Probe of small lathlike fiber
         welter c, me crone associates, inc.
                         236

-------
              ':• Hi'it-'Lc.
                                               (CHRYSOTILE','
  FIBER CONCENTRATION BY NUMBER.. PER  LITER  :
  FIBEP CONCENTRHTION BY MASS.. PER LITER  :
  VOLUME FILTERED  :  100-3.0 ML
  GRID •BClUHP.ES  COUNTED .     40
  TOTAL SUSPENDED  SOLIDS:         0.000  MG  PER LITER
  PH =   0.0
                            0. 42E+05
                            0 . 0 0 1 G R A M 3 ;-: 10 t- •
                         DE': CRI PTIVE  ST AT I 3T I CS

                            HO. OBS.  =     32
  VARIABLE
    MEAN
                                            STANDARD
                                            DEVIATION
                                              ST^MDhRj
                                               Er.SCR
1 LENGTH
2 I.JIDTH
3 ASPECT RrtTIO
4 HrtSS
0. 10305E + 01
0. 33'.:-4iE-01
0.31521E-02
0. lt^47E-01
                           0. 3 5 4 4 is E -^ 0 0
                           0. 1 170'?E-02
                           0.49323E+03
                           0.43S7£E-02
                0.34213E-01    0.
                0 . 2 2 3 2 1 E - y 2    0 . 3'? -
                0.S6033E-01    0.11*
                                                0 4 o   - -'
           Sr'EUNESS
    KURTOSIS
                                             M IN
                                            RANGE
0. 22302E"-0 1
0 . 3 C 3 6 3 E -!• 0 1
0 . 12 ? 4 1 E + 0 1
0.47002E+01
0. is203 lE + 01
0. 3L=73iE + 01
0.llScOE+01
8.21751E-02
0.
0.
0.
M
43J-14E + 01
ISloOE+OO
10143E + -J3
3 o 7 0 0 E + 0 0
                             0. 202'-2IOE-
                             0. 73333EJ-
                             0  2 0 0 0 0 E ~
                                                           yO  0. 4 j?'?
                                                           01  0. 1-"14
                                                           01  0. ?40'?'
                                                           0 3  0 . 3 £ £ 3
                                    237

-------
SAMPLE
— O
*' Q
30
31
"7 •"!•
LE

2
6 .
6 .
0.
1.
1.
0.
0.
0.
1.
0.
1.
1.
0
1.
0.
o .
1 .
0.
0.
0.
0.
|T1 .
1.
o.
4.
-,
0 .
1 .
fl .
1.
1 .
MGTH

0950
3473
4433
2324
3153
2507
'3633
3223
4035
3717
6359
3314
O e t~t "^
4341
3717
2421
6052
2507
3631
3223
9633
4433
6359
1700
3631
3 4 1 4
3645
5245
i i 5>i
4035
291 1
5331
1,!

0
0
0
8
0
o
0
0
O
0
0
0
0
0
0
0
O
o
0
o
0
0
0
0
o
0
0
o
o
0
0
0
IDTH

. 1412
0403
. 0232
. 0232
. 0232
. 0403
. 0232
. 0232
. 8202
. 0202
. 0232
. 0403
. 0605
. 0232
. 0232
. 0202
. 0232
. 0232
0202
. 0202
8202
. 0605
. 0232
0 2 -3 2
. 0403
. 1316
. 0232
0 2 0 2
0403
. 0403
. 0403
. 0202
AS
RH
14.
21.
15.
10.
64.
31.
34.
1 1.
20.
63 .
24.
33 .
20.
17.
43.
12.
21 .
44.
1 3 .
16.
43.
r .
24.
53.
•3
•-, ^
w <_< ,
101.
26.
4-1.
10.
~7'"»
-.' «— .
7* *5
PcCT
7 1 0
3571
0 0 0 0
7143
0000
•"' •'• a; ~?
0 o e o
•-. o cr -'
4236
0 0 0 0
0 0 0 0
2357
0 0 0 0
6667
1429
571 4
0 0 0 0
4236
'"' fi f ~~
0 0 0 0
0 0 0 0
0 0 0 0
£ -^' O -'
-. o =• ~*
0030
r-i n "1 fi
6667
4236
0 0 0 3
0 0 0 0
0 0 0 0
0 0 0 0
0 0 0 0
                                                   0 . 0 9 6 '2
                                                   0 . 0 0 3 2
                                                   0 . 0 0 0 c-
                                                   0. 0005
                                                   0 . 0 0 3 3
                                                   8 . 0 0 4 7
                                                   O. 0013
                                                   6 . 0 0 0 6
                                                   0 . 0 0 0 4
                                                   0. 0013
                                                   8 . 0 0 1 3
                                                   0 . 0 0 5 0
                                                   0 . 0 1 0 5
                                                   0 . 0 0 0 9
                                                   0 . 0 0 2 5
                                                   0 . 0 0 0 2
                                                   O 0 0 1 1
                                                      0 0 0
                                                   8 . 0 0 0 3
                                                   0 . 0 0 0 9
                                                   8. 0037
                    0 0 i 1
                    0 0 1 4
                                                   0 .
                                                   8 .
                                                   8 . 3670
                                                   8 . 0 0 3 3
                                                   8 . 0 0 0 5
                                                   0 . 0 0 6 6
                                                   8 . 0 0 i 5
                                                   0. 0043
                                                   8 . 0 0 1 -
                        238

-------
                         ': M M P L Z"  :


                      DISTRIBUTION B'f LENGTH

    LENGTH             NUN BE,"'      PERCENT         CUMULATIVE
                                                      PERCENT

0 . 0      0 . 3
0.3      1.0
1. n      1.5
1. =      2.0
2. l      2.3
2.3      3.0
3.0      3.3
3.5      4.0
4-0      4.3              .            ,,.v,_,
                                                      100.00
                      DISTRIBUTION  BY  WIDTH

    WIDTH              NUMBER       PERCENT        CUMULATIVE
                                                     PERCENT

3. 0     0. 1             30           93  75             QT  7=;
°- !     0-2              2           6.23            100^00
                    DISTRIBUTION BY rtSPECT  RrtTIO
1
1
y
«'
i
i
0
0
0
1
34,
21 .
•;• cj
|P _
•^' .
T|
0.
0.
0.
3 .
.I1 <
'V I
0 0
J1 1
13
13
90
0 0
0 0
13
1-
:•!
-<-• i
r..t
HI i U
NUMBER
PER
-ENT
CUMULATIVE
PER
-J'
10




^

•-
^
y

o
0




50
60





1
1
1
1

1
1
i
1

^
•f:
i

C1

9
0

M

o
00
10
20
70
"
-
•"
i
c
I
•

•_!
J
Ij
J
J

J
"\ '• ,' ^ ™v




1
10
20
30

40
30
60
70
3 0

90

0 O
1 10
120
130
140
1 ""'""
1
1
1
1

^
2
-i i.i
60
70
3 0
'-* n

'-'-'
00
4
3
-?
r
4
4
1
•?
i
i
—
kl
0
1
0
O
O
0
0
o
o
„
tj
o
o
12
25

21
12
12

^
.}

0
o
•J* .
0 .
o .
o.
0.
0 .
o .
0.

o
o
o.
. 50
00

'"' i
50 '
50
13
'"' *™I
13

0 O
0 0
13
00
00
0 0
0 0
10
0
0

O 0
0 0
0 0


•^








12
%J i

39
7 1
1 i
3-1

'-< ."*!
96

q^r
o^r
-• r-'
1 fi n
100
1 0 0 .
1 00.
1
1
1
1
*

1
1
1
0 0 .
00 .
0 0 .
f ' t"'
~ *
0 0 .
0 0 .
,V-(
LENT
.50
. 50

-.. -^
•— ' i
— -^
30
75
'"' """
'-' '
,— i •" •
'»' i
flf!
0 O
0 0
0 0
0 0
0 0
0 O
0 0

0 0
0 0
f 1 H
                              239

-------
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

-------