EPA-600/1-78-022

  March 1978
                    Environmental Health Effects Research Series
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                RESEARCH REPORTING SERIES

Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology.  Elimination of traditional grouping  was  consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are:

      1.  Environmental  Health Effects Research
      2.  Environmental  Protection Technology
      3.  Ecological Research
      4.  Environmental  Monitoring
      5.  Socioeconomic Environmental Studies
      6.  Scientific and Technical Assessment Reports (STAR)
      7.  Interagency Energy-Environment Research and  Development
      8.  "Special" Reports
      9.  Miscellaneous Reports
This report has been assigned to the ENVIRONMENTAL HEALTH EFFECTS RE-
SEARCH series. This series describes projects and studies relating to the toler-
ances of man for unhealthful substances or conditions. This work is generally
assessed from a medical viewpoint, including physiological or psychological
studies. In addition to toxicology and other medical specialities, study areas in-
clude biomedical  instrumentation and health research techniques  utilizing ani-
mals — but always with  intended application to human health measures.
This document is available to the public through the National Technical Informa-
tion Service. Springfield, Virginia 22161.

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                                   EPA-600/1-78-022
                                   March 1978
PLAN FOR A NATIONAL ENVIRONMENTAL SPECIMEN BANK
                      By
              George M. Goldstein
           Clinical Studies Division
      Health Effects Research Laboratory
     U.S. Environmental Protection Agency
      Research Triangle Park, N.C. 27711
                 Prepared for
        The National Science Foundation
            Washington, D.C. 20550
      HEALTH EFFECTS RESEARCH LABORATORY
      OFFICE OF RESEARCH AND DEVELOPMENT
     U.S.  ENVIRONMENTAL PROTECTION AGENCY
      RESEARCH TRIANGLE PARK, N.C. 27711

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                            DISCLAIMER
     This report has been reviewed by the Health Effects Research
Laboratory, U.S. Environmental  Protection Agency, and approved for
publication.  Mention of trade names or commercial  products does
not constitute endorsement or recommendation for use.
                                n

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                                  FOREWORD
     The many benefits of our modern, developing, industrial society are
accompanied by certain hazards.  Careful assessment of the relative risk of
existing and new man-made environmental hazards is necessary for the estab-
lishment of sound regulatory policy.  These regulations serve to enhance
the quality of our environment in order to promote the public health and
welfare and the productive capacity of our Nation's population.

     The Health Effects Research Laboratory, Research Triangle Park,
conducts a coordinated environmental health research program in toxicology,
epidemiology, and clinical studies using human volunteer subjects.  These
studies address problems in air pollution, non-ionizing radiation, environ-
mental carcinogenesis and the toxicology of pesticides as well as other
chemical pollutants.  The Laboratory develops and revises air quality
criteria documents on pollutants for which national ambient air quality
standards exist or are proposed, provides the data for registration of new
pesticides or proposed suspension of those already in use, conducts research
on hazardous and toxic materials, and is preparing the health basis for
non-ionizing radiation standards.  Direct support to the regulatory function
of the Agency is provided in the form of expert testimony and preparation
of affidavits as well as expert advice to the Administrator to assure the
adequacy of health care and surveillance of persons having suffered imminent
and substantial endangerment of their health.

     This report documents one aspect of an International effort, supported
by EPA, to provide a comprehensive environmental  monitoring program to
assess the relative risk of environmental hazard to the health and well-
being of our population and to aid in the improvement of our environmental
quality.  This program, the National Environmental Specimen Bank, will
serve as an environmental  warning system by providing real  time chemical
analysis of collected specimens.  In addition, this system would permit
the use of tomorrow's more sensitive and more specific methods of chemical
analysis on stored samples.   The advantages of such a program will
permit us to assess the effectiveness of our present environmental
control techniques by monitoring pollutant trends, as well  as establishing
environmental  baseline levels of new pollutants or pollutants of current
concern not previously investigated.
                                      John H. Knelson, M.D.
                                            Director,
                               Health Effects Research Laboratory

                                     iii

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                               ABSTRACT

     The U.S. Environmental Protection Agency (EPA) has been concerned about
the potential dangers to human health and the environment from the ever in-
creasing influx of new man-made substances into our ecosystem.  Since 1974,
the EPA has been actively engaged in studying the feasibility of establishing
a program, The National Environmental Specimen Bank System (NESBS), that
would provide a formalized, systematic approach to assess the environmental
impact of these substances at a national, as well as international level.
The NESBS would provide real time monitoring, assessing the adequacy of pre-
sent pollutant control techniques, as well as providing samples for retro-
spective analyses.
     With the realization that the NESBS is a viable concept, both from the
standpoint of need and by the availability of present technology, it must
be decided if such a system is in fact, feasible to establish and maintain
under actual operating conditions.  It is proposed, therefore, that a five-
year pilot bank program be initiated.  During this time, a limited number of
samples should be collected, analyzed, and stored in a central facility.  The
pilot bank program should focus on validating credible collection, storage,
and analytical data.
     The functioning NESBS should provide future generations with an impor-
tant resource for evaluating their current environmental status.
     This report was submitted in fulfillment of NSF Grant ENV74-19000 A01 by
the U.S. Environmental Protection Agency under the partial sponsorship of the
National Science Foundation, the National Bureau of Standards and the U.S.
Environmental Protection Agency.  This report covers the period June, 1974
through May, 1977.
                                    iv

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                                  FOREWORD
     The many benefits of our modern, developing, industrial society are
accompanied by certain hazards.  Careful assessment of the relative risk of
existing and new man-made environmental hazards is necessary for the estab-
lishment of sound regulatory policy.  These regulations serve to enhance
the quality of our environment in order to promote the public health and
welfare and the productive capacity of our Nation's population.

     The Health Effects Research Laboratory, Research Triangle Park,
conducts a coordinated environmental health research program in toxicology,
epidemiology, and clinical studies using human volunteer subjects.  These
studies address problems in air pollution, non-ionizing radiation, environ-
mental carcinogenesis and the toxicology of pesticides as well  as other
chemical pollutants.  The Laboratory develops and revises air quality
criteria documents on pollutants for which national ambient air quality
standards exist or are proposed, provides the data for registration of new
pesticides or proposed suspension of those already in use, conducts research
on hazardous and toxic materials, and is preparing the health basis for
non-ionizing radiation standards.  Direct support to the regulatory function
of the Agency is provided in the form of expert testimony and preparation
of affidavits as well as expert advice to the Administrator to assure the
adequacy of health care and surveillance of persons having suffered imminent
and substantial endangerment of their health.

     This report documents one aspect of an International effort, supported
by EPA, to provide a comprehensive environmental  monitoring program to
assess the relative risk of environmental hazard to the health and well-
being of our population and to aid in the improvement of our environmental
quality.  This program, the National Environmental Specimen Bank, will
serve as an environmental warning system by providing real time chemical
analysis of collected specimens.  In addition, this system would permit
the use of tomorrow's more sensitive and more specific methods  of chemical
analysis on stored samples.  The advantages of such a program will
permit us to assess the effectiveness of our present environmental
control techniques by monitoring pollutant trends, as well as establishing
environmental  baseline levels of new pollutants or pollutants of current
concern not previously investigated.
                                      John H.  Knelson, M.D.
                                            Director,
                               Health Effects  Research Laboratory

                                     i i i

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                               ABSTRACT

     The U.S. Environmental Protection Agency (EPA) has been concerned about
the potential dangers to human health and the environment from the ever in-
creasing influx of new man-made substances into our ecosystem.  Since 1974,
the EPA has been actively engaged in studying the feasibility of establishing
a program, The National Environmental Specimen Bank System (NESBS), that
would provide a formalized, systematic approach to assess the environmental
impact of these substances at a national, as well as international level.
The NESBS would provide real time monitoring, assessing the adequacy of pre-
sent pollutant control techniques, as well as providing samples for retro-
spective analyses.
     With the realization that the NESBS is a viable concept, both from the
standpoint of need and by the availability of present technology, it must
be decided if such a system is in fact, feasible to establish and maintain
under actual operating conditions.  It is proposed, therefore, that a five-
year pilot bank program be initiated.  During this time, a limited number of
samples should be collected, analyzed, and stored in a central facility.  The
pilot bank program should focus on validating credible collection, storage,
and analytical data.
     The functioning NESBS should provide future generations with an impor-
tant resource for evaluating their current environmental status.
     This report was submitted in fulfillment of NSF Grant ENV74-19000 A01 by
the U.S. Environmental Protection Agency under the partial sponsorship of the
National Science Foundation, the National Bureau of Standards and the U.S.
Environmental Protection Agency.  This report covers the period June, 1974
through May, 1977.
                                    iv

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                               CONTENTS


FOREWORD	 111

ABSTRACT	  1v

FIGURES

     1.  NESB PILOT BANK	  19
     2.  PILOT BANK SAMPLES	  21

  I.  CONCEPTS	   1

 II,  HISTORY	   3

III.  ACCOMPLISHMENTS	  17

 IV.  INTERNATIONAL PROGRAMS	  23

  V.  CONCLUSIONS AND RECOMMENDATIONS	  25

BIBLIOGRAPHY	  26

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I.   CONCEPTS
     Chemicals are an important part of our daily lives.  Like many things,
however, chemicals have both good and bad effects.  Thus, while some chemi-
cals in trace amounts are essential to human life, others can be deadly.
For example, scientists discovered that asbestos and vinyl chloride, two
chemicals commonly used to make a variety of consumer goods, caused
cancer in workers some 20 to 30 years after they were exposed to the
substances.  And recently, consumers learned that chlorine, which is
routinely added to drinking water in many municipalities to kill disease-
causing bacteria, apparently reacts with chemical pollutants in the
water to form barely detectable, but potentially dangerous, amounts of
carcinogenic agents.
     These examples are only two of the discoveries made possible in part
because scientists have developed more sensitive, accurate ways of detecting
and measuring trace chemicals present in very low concentrations in food,
tissues, water and air.  Now people are beginning to wonder which of the
allegedly harmless chemicals in our present day environment might prove to
be health hazards.
     Finding an answer to this question is the major reason that the Envi-
ronmental Protection Agency (EPA), the National Bureau of Standards (NBS)
and the National Science Foundation (NSF) are studying the feasibility of
establishing a National Environmental  Specimen Bank System (NESBS).  The
NESBS would provide a formalized, systematic approach to assess the environ-
mental  impact of these substances at a national as well as an international
level.
     The concepts of the NESBS, real time monitoring and retrospective ana-
lytical capabilities, are derived from its dual function.  First,
                                    1

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representative portions of samples included in the bank would be analyzed
at the time of introduction to provide real time monitoring and evaluation
of pollutant trends. Elevation of these trends would serve as early
warning sentinels so that oroper control measures could be taken to halt
rising human body burdens before irreversible damage could occur.
     Second, a specimen bank would enable measurement scientists to use to-
morrow's more sensitive and specific methods of chemical analysis on today's
samples.  The imoroved measurement methodoloqy would enable health scientists
to determine accurate levels for substances that would be either undetectable
or poorly analyzed by today's less sensitive methodoloqy.  The existence of
a specimen bank would provide the opportunity to determine what the body
burden of newly recognized toxic substances was in the past and to determine
if their levels had changed with time.
     It is anticipated that this formalized, systematic approach, defining
our current environmental hazards, will replace the present system of ran-
domized studies (often without proper validation) used by many State and
Federal regulatory agencies as well as other interested parties in proposing
environmental quality standards and limits for control technology.  If these
types of monitoring programs are to continue, as they must to protect our
environment as well as the health of our population, then it is necessary
to establish and define the basic scientific information required to sustain
such a specimen banking system.  The NESBS, when operational, will provide
future generations with an important resource for evaluating their current
environmental influences.

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II.  HISTORY
     The storage of tissue for analytical measurements is a natural out-
growth of an environmental monitoring system. The need for such a moni-
toring system was realized in the late 1960's by the National Air Pollu-
tion Control Administration, one of the predecessors to the U.S. Environmental
Protection Agency, with the initiation of human population studies.
     The objectives of these studies, (1) to evaluate existing environmental
standards, (2) to quantitate pollutant burdens in exposed populations and,
(3) to quantitate health benefits of pollutant control, were addressed by
coupling sensitive health indicators to comprehensive environmental moni-
toring.  These studies were conducted in communities representing a pollutant
exposure gradient, thus allowing replicated dose response studies over time.
     Environmental monitorinq was divided into two separate aspects, expo-
sure monitoring and tissue pollutant burden monitoring.  Exposure monitorinq
consisted of ambient air monitoring, whereas pollutant burdens, the levels
of environmental residues greater than that required of optimal growth and
development, were measured in selected tissues.  Exposure monitoring was
then correlated with health indicators and covariates to assess the health
impact of the various classes of air pollutants.
     From the very onset of the human pollutant burden studies, short-term
tissue banking became an integral part of this program.  A tissue or groups
of tissues were collected to test a specific hypothesis.   These tissues
were stored until they could be analyzed, and were then discarded.  As the
pollutant burden program expanded,  the need for a fully developed tissue
banking system became more apparent.

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     This banking system was envisioned as having a dual function.  First,
to provide real time monitoring and evaluation of pollutant trends.  Second,
a specimen bank would enable measurement scientists to use tomorrow's more
sensitive and specific methods of chemical analysis on today's samples.
     In EPA's continuing effort to establish a National Environmental Spec-
men Bank, a two day working session was held in February 1973 at EPA's
Rational i-Hvironmental Research Center, Research Triangle Park, North Caro-
lina to discuss and nropose plans for the establishment of a National Envi-
ronmental Specimen BanK System.
     The broad objective? of this working session were:
     1.   Establish current trends in human pollutant burdens (short-term
          banking).
     2.   Create a specimen bank that would provide retrospective analy-
          tical capability (long-term banking).
     One major recommendation from the working session was that a sample
banking system should be established at the national level that would cross
all agency lines and provide human tissues representative of that period in
time from which the sample was taken.  The proper storage of these tissues
would permit retrospective analysis using improved methodologies that were
likely to be available.
     In 1972, the National Academy of Sciences/National Research Council
(NAS/NRC) addressed tissue banking when they stressed the lack of coordi-
nation  in the numerous programs by components of Federal and State govern-
ments,  private industry and academic institutions to collect, store and ana-
lyze specimens of environmental interest.
     In an effort to  upgrade the availability and long-term  protection of
environmental samples and  to make the  information gathered with each

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 II.  HISTORY
     The storage of tissue for analytical measurements is a natural out-
 growth of an environmental monitoring system. The need for such a moni-
 toring system was realized in the late 1960's by the National Air Pollu-
 tion Control Administration, one of the predecessors to the U.S. Environmental
 Protection Agency, with the initiation of human population studies.
     The objectives of these studies, (1) to evaluate existing environmental
 standards, (2) to quantitate pollutant burdens in exposed populations and,
 (3) to quantitate health benefits of pollutant control, were addressed by
 coupling sensitive health indicators to comprehensive environmental moni-
 toring.  These studies were conducted in communities representing a pollutant
 exposure gradient, thus allowing replicated dose response studies over time.
     Environmental monitoring was divided into two separate aspects, expo-
 sure monitoring and tissue pollutant burden monitoring.  Exposure monitorinq
 consisted of ambient air monitoring, whereas pollutant burdens, the levels
 of environmental residues greater than that required of optimal growth and
 development, were measured in selected tissues.  Exposure monitoring was
 then correlated with health indicators and covariates to assess the health
 impact of the various classes of air pollutants.
     From the very onset of the human pollutant burden studies, short-term
 tissue banking became an integral part of this program.  A tissue or groups
 of tissues were collected to test a specific hypothesis.   These tissues
were stored until they could be analyzed, and were then discarded.   As the
 pollutant burden program expanded, the need for a fully developed tissue
 banking system became more apparent.

-------
     This banking system was envisioned as having a dual function.  First,
to provide real time monitoring and evaluation of pollutant trends.  Second,
a specimen bank would enable measurement scientists to use tomorrow's more
sensitive and specific methods of chemical analysis on today's samples.
     In EPA's continuing effort to establish a National Environmental Spec-
men Bank, a two day working session was held in February 1973 at EPA's
National i'r.viror^ental Research Center, Research Triangle Park, North Caro-
lina to discuss arid nropose plans for the establishment of a National Envi-
ronmental Specimen Ban< System.
     The broad objective? of this working session were:
     1.   Establish current trends in human pollutant burdens (short-term
          banking).
     2.   Create a specimen bank that would provide retrospective analy-
          tical capability (long-term banking).
     One major recommendation from the working session was that a sample
banking system should be established at the national level that would cross
all agency lines and provide human tissues representative of that period in
time from which the sample was taken.  The proper storage of these tissues
would permit retrospective analysis using improved methodologies that were
likely to be available.
     In 1972, the National Academy of Sciences/National Research Council
(NAS/NRC) addressed tissue banking when they stressed the lack of coordi-
nation  in the numerous programs by components of Federal and State govern-
ments,  private industry and academic institutions to collect, store and ana-
lyze specimens of environmental interest.
     In an effort to  upgrade the availability and long-term protection of
environmental samples and to make the  information gathered with each

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collection readily available, the Subcommittee on the Geochemical Environ-
ment in Relation to Health and Disease (GERHD) of the National Committee
for Geochemistry, National Academy of Sciences/National Research Council at
their Asilomar Workshop in California (1972) recommended that a group of
specialists be convened to study this problem at its Capon Springs Workshop
in May 1973.
     The concepts from the EPA Specimen Bank Conference were reinforced by
the Capon Springs Workshop (May 6-12, 1973).  The workshop recognized the
need for a NESBS to ensure the continuing availability of a comprehensive
collection of scientifically selected environmental specimens and information.
     The GERHD proposal envisioned the NESBS as a central coordinating insti-
tution, relying on specimens collected from a combination of presently exis-
ting single-purpose collections and materials banked under its own collection
program.  A strict quality control methods standardization program would
ensure that items needed for future environmental studies are acquired, vali-
dated and properly preserved.
     The NESBS could be the mechanism used to continually monitor the envi-
ronment and assess the effectiveness of control practices.  To accomplish
this task, the GERHD Workshop set forth the following objectives:
     1.   Establish mandatory criteria for the sampling, storage, and
          measurement of the various types of specimens that are to be
          accepted into the System.
     2.   Provide historical  specimens for:  (a) the measurement of con-
          taminants not previously investigated, (b) re-evaluation as
          analytical  methods are improved, and (c) measurements of trends.

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     3.    Make  samples  available  for  use  in measuring rates of changes of
          persistent  environmental  substances  (both natural and man-made),
          through  a program  of  systematic sampling and careful storage of
          specimens for retrospective examination.
     4.    Establish a centralized data and information storage and  retrieval
          system appropriate to the scientific community,  and to other inter-
          ested users.
     5.    Provide  samples  that  can  be used in  assessing the long-term envi-
          ronmental effects  of  new industries  and technologies, or  of other
          activities.
     6.    Provide  information useful  for  the assessment of current  environ-
          mental policies, and  for  the establishment of revised environmental
          policies.
     7.    Establish a framework for national coordination  of current and
          future specimen  banks and collection activities  to minimize dupli-
          cation of efforts.
     8.    Foster cooperation and  establish working arrangements for the
          international exchange  of information  and  specimens.
     The GERHD  subcommittee  concluded that the U.S.  Government should esta-
blish, on a permanent basis, a  National Environmental Specimen Banking Sys-
tem.  Initial  coordinating and  funding of the  multi  agency system  should  be
considered by the  National  Science Foundation.   The  Environmental  Protection
Agency should be  considered  as  the most logical  organization  to establish
the system.

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     Four  tasks were proposed to begin development of the NESBS.
          Task  I.   Conduct  an inventory and assessment of the value of
     existing specimen  collections as potential candidates for partici-
     pation  in  the  NESBS.
          Task  II.  Establish a steering committee composed of represen-
     tatives from a variety of concerned groups that are providing funds,
     participating  in specimen collection, and operating monitoring pro-
     grams.  This committee would be responsible for:
               1.   Developing the organizational and managerial structure
                   of  the  NESBS.
               2.   Identifying the types of specimens and information to be
                   stored  in the Banking System.
               3.   Developing interim protocols for sampling, sample handling,
                   and storage of specimens to be included in the bank.
               4.   A plan  for a data handling, storage, and retrieval system.
          Task  III.  Identify research needs as determined during the imple-
     mentation  of the NESBS.  Areas already identified are:
               1.   Sampling strategies
               2.   Sample  processing procedures
               3.   Measurement strategy
          Task  IV.  Conduct meetings at national and international level, of
     user and  research  groups, to .exchange current information that would be
     relevant  to the  NESBS.
     During this time,  the  rapid  growth of EPA's human pollutant burden  pro-
gram dictated  the need  for  a sophisticated system of standardized protocols
for sample collection,  preparation, storage  and  analysis.

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     In December, 1973, this need was addressed by the EPA and the NSF in
a meeting to formulate plans for the development of a National Environmental
Specimen Bank.  At this meeting, a four point proposal was generated, with
joint funding being provided by NSF, EPA and NBS.
     The four point proposal contained the following elements:
     1.   Survey of Existing Specimen Collections
     2.   Evaluation of Specimen Collection Survey
     3.   Research protocol development
     4.   Planning document for the organization and management of the NESBS
1.   Specimen Collection Survey
     The initial chase of the specimen bank program, the specimen collection
survey, was designed to provide a broad data base on the various aspects of
specimen banking.  This data base would subsequently be utilized in deve-
loping guidelines for the NESBS.  Oak Ridge National Laboratory, working
through an interagency agreement with EPA, has conducted a national
survey of existing specimen collections in an attempt to identify those
places in the Continental United States that are currently, or have been
storing material collected  in either research or monitoring activities.
The  survey objectives were  to identify, (1) where collections were located,
 (2)  who maintained the collections,  (3) what the collections  consisted of,
 (4)  what analysis had  been  performed on the material  in the collections,
 (5^  how the sample collections  had  been preserved and stored, and  (6) the
accessibility of the stored materials and associated  data  to  both research
and  regulatory  personnel.   The  survey was designed  to include collections
 in the following areas: geological,  atmospheric, human  tissues,  plant and
 animal tissues  and water  samples.   The  survey  began in  May, 1974 and was
 completed  in  August 1975.   Of the  4506  letters  of survey  intent  that were

                                    8

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mailed, 649 positive responses were compiled into the data base  (Van
Hook and Huber, 1976).
2.   Specimen Collection Evaluation
     NBS has critically evaluated the results of the survey as to their
utility and applicability to the NESBS.  Few of the survey respondents an-
swered the questions in sufficient detail to give a definite answer, but
most of the collections could be of use for taxonomical purposes.
     In developing guidelines for the evaluation of this survey, a large
portion of the recent literature concerning sampling and storage of envi-
ronmental specimens has been examined.  In addition, the advice and opin-
ions of workers in various aspects of the field has been obtained.
     It became readily apparent from the conflicting data in the literature,
that additional research must be conducted in the areas of sample handling
and preparation.  Gross contamination of the samples, from the time of the
sample collection stage, through the analysis stage, appears to be a major
problem (Becker and Maienthal, 1977) and (Maienthal  and Becker, 1976).
3.   Protocol Development
     For a system such as the NESBS to be of any value, the credibility of
the methodology protocols must be above reproach.  EPA realized this condi-
tion at the conception of the NESBS plan, and it has been working with the
National  Bureau of Standards in the development of state-of-the-art method-
ology for sampling, sample handling, analysis and long-term storage.
     It is anticipated that this formalized, systematic approach will  be
invaluable to EPA in defining our current environmental hazards, and in
addition, the methodology protocols would be readily available to other inter-
ested user groups.

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     Since January 1975, the Analytical  Chemistry Division  of  the  National
Bureau of Standards has conducted a continuing  research  program to improve
methodologies for the collection, storage,  and  analysis  of  NESBS samples.
     The program currently underway is pursuing an active research pro-
gram to improve methodologies for sampling, sample handling,  and storage
of biological and environmental  samples for analytical  purposes, as well
as evaluating and improving analytical techniques to be  used  for the ana-
lysis of the trace constituents  of interest.  These portions  of the NBS
research program are currently directed primarily towards trace elements,
but future research will be directed toward other substances  of interest,
such as trace organic species.
     A major portion of the current NBS research program has  been  the
experimental evaluation of contamination and losses of the  trace consti-
tuents of  interest during sampling, sample handling, and long-term storage.
One of the initial projects in this program was the evaluation of twelve
polymeric  materials  for their trace element content, and for the possi-
bility of  removing these trace elements when contacted by liquid samples.
This  study was  made  using three  complimentary  trace analytical  techniques,
neutron  activation analysis,  atomic absorption,  and spark source mass
spectrometry.   The utilization of a multidisciplinary analytical  approach
gave  an  almost  complete coverage of trace  elements  of interest.
      The results of  this  study indicated that  many  materials were  grossly
 contaminated by trace  elements from plasticizers,  formulators,  and other
 process  materials.   However,  conventional  polyethylene  and Teflon were
 found to be  reasonably clean  and it was generally found that  less than
 ten percent  of the bulk trace element content  could be  leached out, even
                                 10

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with conditions as severe as a 2-hour hot leach with 6N^ acid.
     A second, and equally important part of the current research program
has been a study and evaluation of long-term storage techniques which would
be adequate for tissue and other biologically active samples.   The effects
of microbiological action on trace constituent concentrations  and distri-
butions are well documented.  However, the mechanism for complete long-term
elimination of that micro-biological activity is not well documented.
Freezing has long been applied as a technique for analytical  storage, however
no study has yet been performed to document the reliability of this method
of storage for more than a short period of time.
     More recent studies into lyophilization have demonstrated minimal
losses and/or contamination of trace elements during the sample processing.
The NBS has now documented the viability of the freeze-drying  technique to
stabilize trace element composition.  Standard Reference Material Bovine
Liver (SRM1577) has been shown to be unchanged for more than  five years.
The bulk meterial for this SRM was freeze-dried, ground, blended, and
bottled.  This material was analyzed and certified for trace  element compo-
sition in 1972.  To the present time, no documented evidence  of trace element
loss or alteration has occured.
     Finally, the technique of low temperature ashing (LTA) has been evalu-
ated for long-term storage and found to have many advantages.   A recent
study at NBS investigated the loss of trace elements during plasma ashing
using both radioactive tracers, and activation analysis of samples before
and after ashing.  The results obtained indicate that over thirty (30) trace
elements are retained quantitatively during LTA.  Five elements, mercury,
                                 11

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osmium, and the halogens (chlorine, bromine, iodine),  are not quanti-
tatively retained.  It was also determined from the above studies that
contamination of the sample was not a measurable problem during ashing.
An added advantage to the LTA technique is that resultant samples are
easily composited and homogenized.
     The latter portion of the current research program has been to
evaluate the effectiveness, and improve where necessary, the major analy-
tical techniques for environmental samples.   The elements specified to
be of primary interest were mercury, lead, arsenic, selenium, nickel,
vanadium, copper, manganese, beryllium, chromium, platinum, and palladium.
4.   Planning Document for the Organization  and Management of the NESBS
     The formulation of a plan for the development and operation of the
bank will require a series of interrelated tasks.  Some tasks have
been considered in detail, while others are  still in the planning stage.
The actual  completion of the NESBS planning  document will  have to await
the outcome of the Five Year Pilot Bank Program.  The  various tasks
that will be considered in preparing the planning document will  include:
     a.   Review of the Problem:
          An identification and description  of the issues which create the
need for an NESBS.  The kinds of environmental  insults and dangers which
could be better managed and the kinds of analyses required to assess the
magnitude and trends of such dangers will  be described.  In addition, the
contribution that such a bank could make to  better environmental management,
will be estimated.  The result of this task  will be a  statement of need,
including the types of information that would be generated, their various
applications as well as a thoroughly documented rationale for the bank.
                                12

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     b.   Development of Specific Bank Objectives:
          Based on the rationale and need, a set of specific objectives
will be developed to identify just what functions the NESBS will be expected
to perform and how it will meet the needs specified in Task a.
     c.   Identify and Specify Sample Type:
          The types of specimens that will be collected as well as the
areas of collection and trophic levels to be sampled will be based upon
existing and anticipated future needs and objectives as stated in Task b.
Specifications of the kinds of demographic and technological data which
must accompany the samples will be developed.
     d.   Evaluate and Formulate Methods of Analysis:
          Analytical  procedures applicable to the NESBS will be evaluated.
Preferred methodology will be specified along with the amount and condi-
tion of sample required to perform such an analysis.
     e.   Analysis of Specimen Collection, Preparation and Storage
          Requirements:
          Based upon  the kinds and amounts of specimens and the required
storage condition of  samples (so as to maintain their  usefulness) the pro-
cedures available for collection, preparation and storage will  be analyzed
and the preferred methods and procedures will be identified.  In addition,
the total  amounts of  specimens to be stored will be estimated.
     e.   Analysis of Users:
          An identification, analysis and evaluation of potential users
will  be undertaken to provide a set of user specifications as an input to
                                13

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the design of the system and the establishment of operating procedures.
Additionally, a set of guidelines will  be established to govern who can  use
the bank and what may be taken from it.
     g.   Design, Physical  Make-up and Location of the NESBS:
          Tasks b through e will provide the basic information upon which
to establish the functional specifications of the system and will  allow
for the development of a system design.
     The physical make-up will depend substantially upon the numbers and
volumes of specimens, the methods of storage, the analytical instrumentation
and laboratory requirements, and the numbers and kinds of people who will
be provided access to the system.
     The location will depend substantially upon the location of the users,
the location of the sample sites, and to an as yet undetermined degree, upon
what now exists which may become a part of the system.
     h.   Design of a Data Storage and Retrieval System:
          The utility of the NESBS will be expressed by the collection,
maintenance, processing, and dissemination of data.  This data system,
whether an existing off-the-shelf system, or a new set of system speci-
fications will be based upon user requirements and specimen sample re-
aui>-ements.  These requirements will be translated into more meaningful
parameters for the design of the data storage and retrieval system.  These
considerations will include:  data volume and compression for storage,
frequency of user access to data, updating, amending data, data format, data
                                14

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processing, and data traceability.  The NESBS requirements may dictate a
mix of data storage media and associated access systems to be most cost-
effective.  This mix could consist of:
          1.   Photographic storage media (microfilm and microfiche)
          2.   Hard copy volume
          3.   Digital storage media with interactive and non-interactive
               modes
     i.    Develop a Plan for Management and Operation of the Bank:
          The management and operation  plan for the NESBS will  consider the
following:  the maintenance requirements of the physical building(s)  housing
the bank, the storage facility and the  data storage and retrieval  system;
the procedural requirements for the use of the bank, insuring the  economic
utilization by the users; outline of NESBS support requirements; formulation
of a management plan acknowledging the  EPA as the lead agency with unambi-
guous authority for insuring that the NESBS meets its objectives by satis-
fying the user's needs, and recognition of the need to update the  specimen
and data base.
     j.    Budget Plan:
          The budget plan will take into consideration the one-time set
up costs at the inception of the NESBS  and then identify all  of the cost
elements and their contribution to the  total  maintenance costs.

     At the present time, NBS is developing state-of-the-art methodology
for sample collection, preparation, storage and analysis, thus  satisfying
the requirements of tasks d and e.  In  addition, NBS and EPA have  prepared
documentation addressing Tasks a,b,c, and f.   Tasks a,b,c and f are regarded
                                15

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as the statement of need, or feasibility study (Rook and Goldstein, 1977).
The successful completion and acceptance of these tasks would warrant
the completion of tasks g through j.
     The tasks required to develop the NESBS have been designed to give the
maximum amount of information in a step-wise fashion while remaining cost
effective.  Each series of tasks terminates at a GO-NO GO stage, with the
exception of the methodology tasks.  As a result of the urgent need for
methodology protocols and the extended time required for the decision to
develop the NESBS, the methodology protocols will continue until the final
GO-NO GO stage.  It is my opinion that regardless of the outcome of the
NESBS, the methodology protocol development would provide the scientific
community with state-of-the-art standardized protocols for sample collection,
preparation, storage and analysis for a diversity of elements and chemicals
in a variety of ecologically important materials.  The cost benefit of this
alone is uncalculable.
     The initial survey and evaluation task which provided us with the pres-
ent status on specimen collections and sampling, storage and analysis tech-
niques cost approximately $100,000.00.  Protocol development for sampling,
specimen preparation, storage and analysis is. currently costing about
$200,000.00 per year.  The preparation of the NESBS need statement cost
about $50,000.00.
     It is estimated that the remaining tasks g through j may cost about
$300,000.00 to $400,000.00.  These tasks would provide detailed descriptions
for the physical design of a banking system, a data storage and retrieval
system, operation and management plan and an operation budget.
                                   16

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III. ACCOMPLISHMENTS
     On 19 and 20 August, 1976, the NBS and U.S.EPA co-sponsored a workshop
to review technical developments and to make recommendations on the imple-
mentation of the NESBS.  The Workshop consisted of a review session where
past specimen banking considerations were discussed; a technical session
where recent analytical research relevant to sample banking was discussed;
and a planning session where planning and design of a prototype banking
system was outlined.
     On review of the issues that created the need for the NESBS, there
was unanimous agreement that not only was the concept of sample banking
still of vital importance to the assessment of low-level  environmental
contaminiation but that many of the original issues which mandated the
implementation of the NESB were heightened due to recent environmental
pollutant episodes.  The Kepone episode in the James River and the
Chesapeake Bay was pointed to as a prime example were existing specimens
of documented validity would have been extremely useful  to assess the
change in the environment of that pollutant.  There were not and are not
samples of aquatic life or shell fish available from the Chesapeake Bay
or James River which can be used for determination of Kepone levels
before the start-up of the Kepone production in that area.  Limited samples
were available from the Virginia Institute of Marine Sciences dating back
3-4 years.  These samples have proven invaluable in establishing the
extent to which Kepone has affected the marine life.  Had earlier samples
been available from a banking system, a far better assessment of Kepone
baseline levels prior to the dumping episodes would have been available
                                17

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to environmental  officials.   With the large increase of man-made chemicals
now being put into our environment, it is evident  that  issues  such  as
the Kepone insult in Virginia are surely to be on  the increase rather
than to remain as an isolated situation.
     The single most important point to come from  the Workshop was  that the
NESBS can serve many important functions, not just that of long-term retro-
spective analysis.  The results of sample banking  would surely impact  on
monitoring and health effects research of the EPA, and  would be of  great
assistance to ongoing programs within the Department of Agriculture,
Food and Drug Administration, and other agencies.
     A set of specific objectives for the sample bank were identified.
(Figure 1)  Those objectives are summarized as follows:
     1.   The collection, preservation, and storage of selected environmental
          samples using methodologies that had been documented to minimize
          or eliminate alteration of trace constituents.
     2.   The real-time analysis of selected trace constituents using
          methods of documented validity to obtain monitoring trend
          data.
     3.   Research in analytical methodology utilizing both the accumu-
          lated long-term data base and samples that have been stored  in
          the NESBS.  This research will lead to a self-improving set  of
          monitoring data.
     4.   The periodic review of the operation of the banking system
          relative to its valid input of samples and output of analytical
          data.
                                18

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 RESEARCH IN
 ANALYTICAL
METHODOLOGY
                   NESB ADMINISTRATION
 REAL TIME
MONITORING
LONG TERM
 STORAGE
     RESEARCH IN
SAMPLE PRESERVATION
     AND STORAGE
                                       \
          RESULTS
      TIME SERIAL DATA
     FOR TREND ANALYSIS
                     RESULTS
                   SAMPLES FOR
             RETROSPECTIVE ANALYSIS
                   QUALITY ASSURANCE
                    TIME DIFFERENTIAL
                       COMPARISONS
                     Figure 1.  NESB PILOT BANK
                                19

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     The conclusions expressed by the Workshop  participants was  that  a
formalized, systematic approach is needed to  assess  the  environmental impact
of the ever increasing influx of man-made substances into  our  ecosystem.
The technology to initiate this type of program is  presently available  and
should be formulated into a pilot bank program.   This program  would  utilize
and integrate our newly developed techniques  with those  currently used.
The pilot bank program would serve as the "test site" for  program design
and implementation.  The various aspects of the program  would  be evaluated
at each stage of development.  The knowledge gained from operating this
pilot study would be incorporated into the NESBS plan.
     The identification of specific sample types which should  be included
in the pilot sample bank during initial start-up of the  system was
discussed. The major focus of attention was on  the absolute  requirement
to minimize both the number of samples and sample types  in the pilot
program so that the banking system did not become overwhelmed  with
either samples or analyses during its first years of operation.   Unreason-
ably large numbers of avoidable errors would destroy its credibility
before it was even in full operation.
     Thus, all participants recommended the inclusion of a modest sample
set per year for the first five years of operation with the focus on
validating credible storage and analytical results.
     A reasonable figure of approximately two thousand samples a year,
split into four matrix types, was recommended (Figure 2).   Also, it was
                                 20

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           TWO GEOGRAPHIC  LOCATIONS
               2000 SAMPLES PER YEAR
 HUMAN TISSUE
ACCUMULATOR
ATMOSPHERIC
  INTERGRATOR:
  LICHEN, MOSS,
   FILTERS
 FOOD
GRAIN
                AQUATIC
            ACCUMULATOR
          SHELLFISH BIVALVE
             Figure 2.  PILOT BANK SAMPLES
                           21

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recommended that the samples  be  obtained  from  no more than two different
locations in the beginning phases,  to minimize problems with  logistics,
sample collection,  and transport.
     Nine different sample types representing  all major phases of  environ-
mental samples were considered for  inclusion in the  bank.  These included
air particulates, sediment, water,  botanical,  biological, and human
samples. It was recommended that samples  which represented environmental
accumulators or integrators be emphasized as initial candidates, both
due to the ease of analytical manipulation of  those  samples and because
they represented time integrators of major pollutants present in our
environment.
     The first sample that was recommended for inclusion  unanimously was
a soft tissue sample that had an accumulator function in  the  human body,
most likely liver or kidney.   The second  and equally important  sample
type was an accumulator of aquatic  origin. Agreement was reached  that a
shellfish bivalve such as oyster which  passes  large  quantities  of  water
through its system every day  and which  tended  to  mirror  increased  concen-
trations of many toxic pollutants was  a good choice. The third  sample
type was a food material representing  a major  input  into  the  human diet.
Consensus was unanimous that  a food grain or composite  of grains was  the
best choice.  The fourth sample type was  a collector of atmospheric or
airborne pollutant materials. It was recommended  that material  such a
lichen or moss was a good  indicator of long-term  trends  in  atmospheric
pollutants and should be included in the initial  bank.   These four
sample types were chosen for their diversity  and  their  utility  to  environmental
                                22

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           TWO GEOGRAPHIC LOCATIONS
              2000 SAMPLES PER  YEAR
 HUMAN TISSUE
ACCUMULATOR
ATMOSPHERIC
  INTERGRATOR:
  LICHEN, MOSS,
   FILTERS
 FOOD
GRAIN
                AQUATIC
            ACCUMULATOR
         SHELLFISH BIVALVE
             Figure 2.  PILOT BANK SAMPLES
                           21

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recommended that the samples  be  obtained  from  no more than two different
locations in the beginning phases,  to minimize problems with logistics,
sample collection,  and transport.
     Nine different sample types representing  all major phases of  environ-
mental samples were considered for  inclusion in the  bank.  These included
air particulates, sediment, water,  botanical,  biological, and human
samples. It was recommended that samples  which represented environmental
accumulators or integrators be emphasized as initial candidates, both
due to the ease of analytical  manipulation of  those  samples and because
they represented time integrators of major pollutants present in our
environment.
     The first sample that was recommended for inclusion  unanimously was
a soft tissue sample that had an accumulator function in  the human body,
most likely liver or kidney.   The second  and equally important  sample
type was an accumulator of aquatic  origin. Agreement was reached  that a
shellfish bivalve such as oyster which  passes  large  quantities  of  water
through its system every day  and which  tended  to  mirror  increased  concen-
trations of many toxic pollutants was  a good choice. The third sample
type was a food material representing  a major  input  into  the  human diet.
Consensus was unanimous that  a food grain or composite of grains was  the
best choice.  The fourth sample type was  a collector of  atmospheric or
airborne pollutant materials. It was recommended  that material  such a
lichen or moss was a good indicator of long-term  trends  in  atmospheric
pollutants and should be included in the initial  bank.   These  four
sample types were chosen for their diversity  and  their  utility  to  environmental
                                22

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monitoring programs.   However, a second and even more important consideration
was that there is now enough scientific evidence to be reasonably assured
that the storage and analysis of the trace element components in these
materials could now be carried out with integrity (Rook and Goldstein,
1977).
     One of the last items for consideration at the Workshop was the
evaluation and formulation of  viable analytical methods for the real-
time and retrospective analysis of samples from the NESBS system.  Part
of the NBS research effort has been to publish a compilation of analytical
methods currently being used for SRM certification analysis.  These
methods will  be available in the near future.
     The concept of specimen banking to assess the environment was rein-
forced at an International Workshop "The Use of Biological  Specimens
for the Assessment of Human Exposure to Environmental Pollutants" held in
Kirchburg, Luxembourg, 18-22 April, 1977 (Goldstein, 1977).
     The conclusions and recommendations from this Workshop stated that
where the scientific expertise is already available, that biological moni-
toring should be instituted to provide direct evidence of human exposure to
chemicals in the environment.  In addition, research should be instituted in
those areas where the scientific expertise is lacking or insufficient.
IV.  INTERNATIONAL PROGRAMS
     The problems of environmental pollution are worldwide in scope and
transgress all national and political boundaries.  Finding answers to
problems of environmental concern in areas of pollutant identification,
sources, effects, control, etc. are the concerns of many nations.
                                23

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     Since 1975, the United States Environmental  Protection Agency and
the Environmental Agencies of the Federal  Republic of Germany (FRG) have
been pursuing collaborative research in areas of the tissue bank program as
part of a bilateral agreement on the environment.  This cooperative program
allows for the input of expertise from the two countries, thus reducing
research time and cost.  During this time period, the U.S. effort has focused
on research areas related to trace elements, whereas, the FRG has concen-
trated their efforts in the area pertaining to organic substances.
     Meetings are held on a yearly basis to discuss and exchange research
findings.
                                    24

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V.   CONCLUSIONS AND RECOMMENDATIONS
1.   The National Academy of Sciences/National  Research Council  stressed
     the lack of coordination in the numerous programs by components of
     Federal and State government, private industry and academic institu-
     tions to collect, store and analyze specimens of environmental  interest.
2.   A formalized, systematic approach, defining our current environmental
     hazards, must replace the present system of randomized studies  (often
     without proper validation) so as to protect our environment as  well
     as the health of our population.
3.   It is necessary to establish and define the basic scientific information
     required to sustain a specimen banking system.
4.   The conclusions and recommendations of two workshops (NBS/EPA - Aug. 1976
     and WHO/CEC/EPA - Apr. 1977) proposed that where the scientific expertise
     is already available, that biological monitoring should be instituted  to
     provide direct evidence of human exposure to chemicals in the environment.
                                  25

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                            BIBLIOGRAPHY
Van Hook, R.I. and Huber, E.E.  National Environmental Specimen Bank Survey
  (1976) U.  S. Environmental Protection Agency Publication #EPA-600/1-76-006.

Becker, D.A. and Maienthal, E.J.  Evaluation of the National Environmental
  Specimen Bank Survey. (1977)  U. S. Environmental Protection Agency
  Publication #EPA-600/l-77-015.

Maienthal, E.J. and Becker, D.A.  A Survey of Current Literature on
  Sampling,  Sample Handling, and Long term Storage for Environmental
  Materials. (1976)  National Bureau of Standards  Publication #NBS-TN-929.

Rook, H.L. and Goldstein, G.M.  Recommendations of the EPA/NBS Workshop
  on the National Environmental Specimen Bank.  (1977)  U. S. EPA
  Publication #EPA-600/1-77-020.

Goldstein, G.M.  The National Environmental Specimen Bank; Its Concepts,
  History and Objectives. International Workshop on "The Use of Biological
  Specimens for the Assessment of Human Exposure to Environmental Pollutants.
  April 18-22, 1977, Luxembourg.  Sponsored by WHO, U. S. EPA, and CEC.
                                  26

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                                  TECHNICAL REPORT DATA
                                readluidj'uctioiU: on the reietse before com/;/.
1  REPORT NO.
  EPA-600/1-78-022
4. TITLE AND SUBTITLE
 PLAN FOR A NATIONAL  ENVIRONMENTAL SPECIMEN BANK
                                                          6. PERFORMING ORGANIZATION CODE
                                                          3 RECIPIENT S ACCESS'O> NO
              5. REPORT DATE

                March 1978
7 AUTHOR(S)
                                                          8. PERFORMING ORGANIZATION REPORT NO.
 George M. Goldstein
9. PERFORMING ORGANIZATION NAME AND ADDRESS
 Clinical Studies  Division
 Health Effects  Research  Laboratory
 U.S. Environmental  Protection  Agency
 Research Triangle Park,  NC 27711
              10 PROGRAM ELEMENT NO.

               1AA601  	
              11. CONTRACT/GRANT NO.
12. SPONSORING AGENCY NAME AND ADDRESS
 Health Effects  Research  Laboratory
 Office of Research  and  Development
 U.S. Environmental  Protection Agency
 Research Triangle Park,  N.C.  27711
                                                          13. TYPE OF REPORT AND PERIOD COVERED
RTP,NC
              14. SPONSORING AGENCY CODE
               EPA 600/11
15. SUPPLEMENTARY NOTES
16. ABSTRACT
      The U.S.  Environmental  Protection Agency (EPA) has  been  concerned about the
 potential dangers  to  human  health and the environment from  the  ever increasing flux
 of new man-made  substances  into our ecosystem.  Since 1974, the EPA has been actively
 engaged in studying the  feasibility of establishing a program,  The National  Environ-
 mental Specimen  Bank  (NESB),  that would provide a formalized, systematic approach
 to assess the  environmental  impact of these substances at a national,  as well  as
 international  level.   The  NESBS would provide real time  monitoring, assessing the
 adequacy of  present pollutant control techniques, as well as  providing samples for
 retrospective  analyses.
      With the  realization  that NESBS is a viable concept, both  from the standpoint of
 need and by  the  availability  of present technology, it must be  decided if such a
 system is, in  fact, feasible  to establish and maintain under  actual operating
 conditions.  It  is proposed,  therefore, that a five-year pilot  bank program be
 initiated.   During this  time, a limited number of samples should be collected,
 analyzed, and  stored  in  a  central facility.  The pilot bank program should focus on
 validating credible collection, storage, and analytical  data.   The functioning NESBS
 should provide future generations with an important resource  for evaluating their
 rurrpnt.
17.
                               KEV WORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
 data acquisition
 Information centers
 Environmental  surveys
                                             b.IDENTIFIERS/OPEN ENDED TERMS
   specimen bank
                              COSATi Field/Group
05 B
06 F
13, DISTRIBUTION STATEMENT

 RELEASE TO PUBLIC
 19 SECURITY CLASS /Th.s Report;


 201?ECCttfTY C~LWss '/Thisjage'l
                                                                        21 NO OF PAGES
                                                                        22 TRICE
                                               UNCLASSIFIED
EPA Form 2220-1 (9-73)
                                            27

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