United States
                   Environmental Protection
                   Agency
Office of Health and
Environmental Assessment
Washington DC 20460
                   Research and Development
EPA/600/S8-85/020 Aug. 1986
s>EPA         Project  Summary
                   Evaluation  of Methods for
                   Analysis  of Human  Fat,  Skin,
                   Nails,  Hair,  Blood,  Urine  and
                   Breath
                   L Sheldon, M. Umana, J. Bursey, W. Gutknecht, R. Handy, P. Hyldburg,
                   L Michael, A. Moseley, J. Raymer, D. Smith, C. Sparacino, and
                   M. Warner
                    This research program surveyed and
                   evaluated the methods and procedures
                   used to identify and quantitate chemi-
                   cal constituents in human tissues and
                   fluids including fat, skin, nails, hair,
                   blood, urine, and breath. These meth-
                   ods have been evaluated to determine
                   their ease and rapidity, as well as cost,
                   accuracy, and precision. During this
                   evaluation, a second goal was to deter-
                   mine the feasibility of correlating a pre-
                   ferred method with a specific tissue/
                   fluid and with easily identifiable
                   chemical and physical characteristics of
                   the analyte.
                    Because of these goals, the search
                   strategy, as well as the literature evalu-
                   ation focussed on analytical methods.
                   The literature search was restricted to
                   lists of chemicals of current interest to
                   the U.S. Environmental Protection
                   Agency, to references  cited in
                   "Chemicals Identified in Human Biolog-
                   ical Media, A Data Base," and the
                   "Chemical Abstracts Data Base." The
                   information retrieved was summarized
                   and classified by sampling and analysis
                   methodology.
                    This Project Summary was devel-
                   oped by EPA's Office of Health and En-
                   vironmental Assessment, Washington,
                   DC, to announce key findings of the re-
                   search project that is fully documented
                   in two separate reports (see Project Re-
                   port ordering information at back).
Introduction
  For a long time, air monitoring consti-
tuted the major means of assessing
workers' exposure to chemicals in in-
dustry. But this monitoring takes into
account only exposure via the pul-
monary route and, even for respirable
chemicals, it  does not indicate the ac-
tual uptake by the exposed worker.
  Biological  monitoring, the routine
analysis of human tissues or excreta for
direct or indirect evidence of exposure
to chemical substances, has also been
used to learn more about early  detec-
tion of health impairment due to  indus-
trial chemicals.  The types of analyses
include the following measurements:
  • concentration of the chemical in
   various  biological  media such as
   blood, urine, tissue and hair;
  • concentration of metabolites of the
   original chemical in the same me-
   dia; and
  • determination  of adverse/nonad-
   verse biological changes of the or-
   ganism resulting from exposure to
   reaction  of the organism to expo-
   sure.
  Hence, biological  monitoring is used
in the assessment of human exposure.
A main goal  of such monitoring is to
ensure that the current or past levels of
worker exposure are safe. Biological
monitoring is being applied increas-

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ingly to the assessment of exposure in
environmentally exposed nonworker
populations.
  The purpose of this research program
was to survey and evaluate the methods
and procedures used to identify and
quantitate chemical constituents in hu-
man tissues  and fluids  including fat,
skin, nails, hair, blood, urine, and
breath. These methods have been eval-
uated to determine their ease and rapid-
ity, as well as cost, accuracy, and preci-
sion. During this evaluation, a  second
goal was to determine the feasibility of
correlating a  preferred method with  a
specific tissue/fluid and with easily
identifiable chemical characteristics,
such as octanol-water partition coeffi-
cient, water solubility, vapor pressure,
etc.
  Because of these goals, the search
strategy and the literature evaluation fo-
cussed on analytical methods. The liter-
ature search was restricted to  lists of
chemicals of current interest to the U.S.
Environmental Protection Agency
(EPA), to references cited in "Chemicals
Identified in Human Biological Media, A
Date Base" up to 1982  and the
"Chemical Abstracts Data Base." The
Chemical Abstracts  Data Base covers
the complete chemical  literature from
1967 to present, however, the search
performed for this review was restricted
to 1976 to 1983.
  The list of target chemicals included
in this review was compiled from a set
of five documents received  from the
EPA Technical Project Monitor. The doc-
uments contained lists of compounds of
interest to the EPA.
  The chemicals were classified into a
hierarchial scheme based on their phys-
ical and chemical properties. This
scheme first separated  inorganic from
organic compounds. Organic chemicals
were then separated according to func-
tional group  substituents.  Functional
groups that have the greatest effect on a
compound's physical and  chemical
properties and/or analytical behavior
are placed highest in the scheme. For
example, carboxylic acids are ranked
above acid esters. The acid moiety is a
stronger functional group with a greater
influence on aqueous solubility, boiling
point,  extraction behavior, and chro-
matographic properties. If a compound
contains more than  one functional
group, it is placed in the highest chemi-
cal classification. Because this  classifi-
cation is based on the physical/chemical
properties, it should reflect analytical
behavior of many of the compounds to
aid in the overall evaluation process.
  Information from the data  base
"Chemicals Identified in Human Biolog-
ical Media" on (1) literature referenced,
(2)  chemicals  analyzed, (3) biological
matrix, and (4) analytical technique was
entered and stored on computer files.
These data were retrieved according to
analytical method. Under each analyti-
cal  method, citations  referring to that
method were listed according to chemi-
cal class, specific chemical, and biologi-
cal matrix. The "Chemical Abstract Data
Base" was searched using the selected
compounds which were  cross-
referenced by CAS number with the
human  biological matrices of interest.
This information was  used for sorting
retrieved literature citations and dis-
seminating articles to members of the
advisory committee who were  respon-
sible for the preparation of this review.
This information was also  used to
match analytes with biological matrix in
order to determine the feasibility of cor-
relating analytical method with  biologi-
cal matrix and with chemical character-
istics.
  Relevant articles which report the use
of an analytical technique to measure
any of the target chemicals in the seven
biological matrices of interest were ob-
tained. Only those analytical methods
which had at least three  articles cited
v jie included in the review.
  During acquisition of literature cita-
tions, more than 95% of all citations
identified as relevant,  i.e., those which
reported measuring any of the target
chemicals in the seven biological ma-
trices, were retrieved. Missing citations
were not retrieved for  several reasons:
  • the article was in a foreign lan-
    guage which could not be trans-
    lated easily;
  • the reference in the data base was
    incorrect; or
  • a source of the article could not be
    located.
  The full report was written to evaluate
each analytical method. The evaluation
for  each method  included sections  on
instrumentation  and  sample prepara-
tion methods. Much of the information
on analytical instrumentation was taken
from review articles, reference texts, or
our working knowledge  of the  tech-
niques. Information on sample prepara-
tion methods were generally restricted
to the citations in the data bases. The
section  on  method/analyte correlation
parameters attempted to suggest physi-
 cal or chemical  properties of analytes
 which could be used to predict the ap-
 plicability of the given method to the
 analysis of that  chemical in a specific
 biological matrix.

 Conclusions
  Methods and  procedures that were
 cited in both data bases for the identifi-
 cation and quantitation of the target
 chemicals in the  seven human biologi-
 cal  matrices were  summarized. Broad
 generalizations are difficult to formulate
 due to the number of compounds, com-
 plexity of matrices, and the variety ol
 methods reviewed. A noncritical survey
 of the analytes for each method in the
 data base with a brief summary of the
 advantages and disadvantages of each
 of the methods reviewed follows.

Atomic Absorption Spec-
 trophotometry
  Atomic  absorption spectrophotome-
try  has  been the method of choice foi
 many  investigators who  require c
 highly specific, low cost technique foi
elemental analysis with ultratrace  de
tection limits. The advantages of this
single-element  method have beer
 known for the past 15 years, but the use
 of modern instrumentation has pro
duced analytical  data of greater repro
ducibility and has resulted in increasec
sample  throughput.  One of the  best
documented environmental measure
 ments (lead in whole blood) is usualh
performed by  atomic  absorptior
methods.
  The biggest disadvantage of this tech
nique is not the inherent limitations o
the method with  respect to analyt
specificity and sensitivity, but problem
with matrix effects. A variety of diges
tion conditions  have been developei
which in combination with  deuteriur
 background correction  and/o
chelation-extraction, such  as APDC
 MIBK, have been  successful in reducin
matrix effects and form the basis fc
most atomic absorption procedures.

 Neutron Activation Analysis
 (NAA)
  Neutron activation allows  multiel
 ment analyses on a single sample wit'
 out pretreatment.  Under conditions <
 high neutron flux, maximum samp
 size (approximately 1 g) and no interfe
 ences, NAA provides detection limi
 which can only be matched by flam
 less atomic absorption spectrophot

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metry. Essentially all of the elements of
environmental interest (with the excep-
tion of beryllium and lead) from nitro-
gen to the heavy elements can be ana-
lyzed simultaneously by this technique.
  The principal  limiting factor in neu-
tron activation analyses is the presence
of interferences that  result  in the
gamma-ray signal from the various
sources not being resolved. Detection
limits may increase by several orders of
magnitude  when interferences are
present. Poorer precision and accuracy
also result.
  Accessibility  to a  neutron source
notwithstanding, NAA can be relatively
simple and inexpensive with the capa-
bility of measuring a large number  of
elements simultaneously. When  either
chemical methods or extensive time
resolution is required to minimize inter-
ferences; the cost, time,  and labor re-
quired for analysis increase signifi-
cantly.


Electrochemical Methods
  The facility of ion selective  poten-
tiometry in  the analysis of inorganic
ions is a distinct advantage of this ana-
lytical technique. Although limits of de-
tection are typically  10~6 M, which  is
somewhat higher than competing tech-
niques, fluoride and cyanide selective
electrodes have been quite successfully
used in the analysis of blood and urine.
Their simplicity  in this  regard  makes
them highly recommended.
  Anodic stripping voltammetry  (ASV)
has been applied to the analysis  of
blood, urine, and hair for a large num-
ber of metals including zinc, cadmium,
lead, chromium, and copper. Since ASV
is capable of differentiating between
labile and nonlabile metal species, it is
clearly the method of choice when infor-
mation regarding metal  speciation  is
desirable. When total metal determina-
tions are needed, ASV is comparable to
other methods with respect to labor re-
quired and accuracy.
  Cyclic voltammetry has been applied
to several metal  determinations and, in
addition, some  organic compounds
such as p-amino-diphenylamine. Cyclic
voltammetry is an easy technique to im-
plement and the  analysis is fast;  blood
samples  have been  analyzed without
further treatment. The equipment cost
is relatively  inexpensive  compared  to
other instruments used in the simulta-
neous determination of metals. The
limit of detection (LOD) is higher than
for ASV.
Emission Spectroscopy
  A large number of metals may be de-
termined using emission spectroscopy.
The limit of detection for these metals is
adequate for environmental trace analy-
sis, and many published data were
found on the use of either DC or argon
plasma  ionization methods. However,
by far, the most widely used technique
is atomic absorption (AA) spectroscopy.
Because of the wealth  of data on
method performance and the availabil-
ity of instrumentation, AA would be
considered the method  of first choice,
but ICP  may be faster, more accurate,
more precise and have lower LOD.

Proton Induced X-Ray Emission
(PIXE)
  Proton induced X-ray analysis may be
used for multielement analysis  of a
large number of samples. Although bio-
logical samples can be analyzed  in a
variety of forms, including untreated
specimens, best results are achieved by
digesting the organic material and  plac-
ing the  resulting solution on a Mylar
film for irradiation. The elements nor-
mally measured  for PIXE range from
phosphorous  (2= 15) to lead (2 = 82)
with limits of detection  at the  sub-
nanogram level.
  The method is simple,  allows  high
sample throughput, and  is currently be-
ing performed inexpensively by several
laboratories.

Spectrophotometry (Colorime-
try)
  Spectrophotometry has  the advan-
tage of being a simple and quick proce-
dure utilizing instrumentation which is
widely available and relatively inexpen-
sive. It is capable of determining com-
pounds and metal complexes at the low
microgram to high nanogram range
with good precision (2-3%). The disad-
vantages of this technique are the labor
intensive sample preparation steps, and
the nonunique nature of the measured
signal (compared to AA/ICP).  This
method is suitable for many analytical
measurements, particularly where the
operating laboratory does not have AA/
ICP spectrometers. When available, AA
or ICAP would usually be the methods
of choice.

Gas Chromatography
  Gas Chromatography  is  used for the
separation and determination of volatile
or semivolatile organic compounds. Re-
cent developments in capillary column
Chromatography provide for very high
resolution to give separation of com-
plex samples. The use of specific
column packings allows the determina-
tion of polar and some compounds with
low volatility.
  Effluent from the GC column may be
analyzed using a specific (e.g., electron
capture, nitrogen/phosphorus) or a non-
specific (e.g., flame ionization, thermal
conductivity)  detector. Limits of detec-
tion range from subpicogram to micro-
gram depending upon the detector
used and the compounds determined.
  Gas Chromatography is a simple tech-
nique for the determination  of organic
compounds and is used extensively on
human biological extracts. Compounds
which  are  nonvolatile  (B.P. >300°C),
very polar,  or heat labile cannot be de-
termined directly by this method. For
these compounds, high performance
liquid  Chromatography is  generally
used. Some  polar and/or nonvolatile
compounds may be derivatized to im-
prove gas  chromatographic perform-
ance.
  Most biological samples  cannot be
analyzed directly. Analytes are  usually
extracted and fractionated prior to anal-
ysis. Although GC analysis  itself is  a
simple rapid procedure, sample prepa-
ration techniques are often complicated
and time-consuming.
Mass Spectrometry
  Mass spectrometry  is a  sensitive,
specific but expensive technique.  The
various forms of mass spectrometry
that  can  be employed  complicate the
question  of when the technique should
be used. Spark source mass spectrome-
try is extremely valuable in elemental
analysis. Atomic absorption  spec-
trophotometers and inductively cou-
pled argon plasma  spectrometers are
substantially  less expensive for these
analyses; however, for a semiquantita-
tive scan of a  broad range of elements,
spark source mass spectrometry could
be the method of choice. Many com-
mercial laboratories offer spark  source
mass spectrometric assays as a service,
so purchase would not be necessary for
an occasional need. The main  use of
mass spectroscopy is in the  identifica-
tion and  quantitation of organic com-
pounds.
  Isotope ratio mass spectrometry is
the preferred  method for accurate de-
termination of isotope ratios. If such an
assay is required, specialized laborato-
ries make effective use of the  instru-
mentation.

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  Although gas chromatography/mass
spectrometry as a combined technique
is one of the most widely used analyti-
cal  methods, the decision to use the
technique should be evaluated care-
fully. Gas chromatography is far more
cost-effective and is usually more sensi-
tive. However, the analysis of biological
samples for various  organic con-
stituents by gas chromatography pre-
sents various difficulties which may re-
quire the  specificity inherent  in the
mass spectrometric technique. For ex-
ample, if identification of a pesticide is
desired  for forensic or regulatory pur-
poses, the use of gas chromatography/
mass spectrometry with authentic
standards will give the needed informa-
tion. Biological samples often contain
material which  coelutes with  a com-
pound  of interest and only the
specificity of the  mass spectrometric
technique will give a successful assay.
When standards for the compounds  of
interest  are not available, identification
can be carried out with a high degree of
confidence  only by the mass spectro-
metric technique.
  In summary, mass  spectrometry
should be used when its specificity is
required, a  less expensive technique is
not adequate, or when certain types  of
information are available from no other
source,  for example,  accurate isotope
ratios or accurate mass measurement.

X-Ray  Spectroscopy
  X-ray  emission spectroscopy can be
used in chemical analysis for both quan-
titative and qualitative  studies.  X-ray
spectroscopy is applicable to the deter-
mination of most metals in multiele-
ment analysis. The technique is expen-
sive and not widely available.

Nuclear Magnetic Resonance
(NMR)
  Nuclear magnetic resonance  spec-
troscopy is applicable to a wide range of
organic  compounds, provides informa-
tion on chemical structure, and can be
quantified.  The  operation of NMR  in-
struments is rather complex; however,
no significant sample preparation is re-
quired. The equipment  is very expen-
sive and not widely available.

Thin Layer Chromatography
  Thin layer chromatography is a sepa-
ration technique based  on an adsorp-
tion or partition process and employs a
thin, flat bed of sorbent. Thin layer chro-
matography can be used for the separa-
tion and semi-quantitation of complex
mixtures and has. proven useful as a
cleanup technique for analvtes present
in biological fluid extracts and environ-
mental samples.
  With the exception of some macro-
molecules, reactive compounds, and
volatile substances; virtually  any or-
ganic compound and many.inorganic
ions can be detected by thin layer chro-
matography. The limit of detection is a
function of the specific .detection sys-
tem utilized. Semi-quantitative thin
layer  chromatography  is fast, simple
and can be accomplished with a mini-
mum  of expensive equipment. Quanti-
tative thin layer chromatography with
modern instrumental detection systems
is both  accurate  and sensitive but re-
quires rather expensive  equipment.

High Performance Liquid Chro-
matography
  High performance liquid chromatog-
raphy is a simple and standard separa-
tion technique applicable to virtually
any mixture of organic  compounds as
well as certain ionic species. It is usually
applied to complex matrices for which
gas chromatography is not usable, such
as  matrices containing high  boiling
point  chemicals or compounds which
decompose under GC  conditions. A
wide variety of detectors can be inter-
faced to  a liquid chromatography
column such as ultraviolet detectors, re-
fractive index and fluorimetric detec-
tors.

Miscellaneous Methods
  During this survey, two reports were
found dealing with the determination of
compounds of interest using some mis-
cellaneous methods. The scarcity of
publications did not warrant a special
section for these  techniques in this re-
view.
  One such report describes the evalua-
tion of the permeability of phenolic
compounds through human stratum
corneum using a desorption technique.
In another report, laser microprobe
mass analysis (LAMMA) is used to de-
termine metal ions and organic com-
pounds in solid samples which require
spacial resolution.

Methods for Breath Analysis
  Several  combinations of sampling,
concentration, and analysis methods
were cited in the reviewed literature for
the identification  and quantitation of
target compounds in human breath. A
brief noncritical review of the  advan-
tages  and disadvantages of each
method follows.
  In 1977 an international workshop or
"The Use of Biological Specimens foi
the Assessment of Human Exposure tc
Environmental Pollutants" summarizec
the data on the location or compartmen
talization  of xenobiotics  and/or thei
metabolites in various biological ma
trices including breath. They concludec
breath  is not useful to evaluate expo
sure to arsenic, beryllium, cadmium
chromium, fluorine,  lead, mercury
molybdenum, manganese, nickel, vana
dium, zinc, copper, cobalt, platinum
palladium, tin, methyl  mercury, DDT
phenoxy herbicides, pentachlorophe
nol, PCB, PBB,  PCN, tetrachloro
dibenzo-p-dioxins, benzofuran, azoben
zene, polycyclic hydrocarbons, amini
and nitro derivatives, organophospho
rus esters, cyanides, nitriles, mycotox
ins, antibiotics, and  hormonal sub
stances. They concluded breath level:
correlate with body burden in the case:
of  chlorinated  solvents, plasti
monomers, fluorinated propellants
nonsubstituted aliphatic and aromati
volatile hydrocarbons, alcohols,  ethers
ketones, and carbon monoxide. It wa
also concluded selenium breath level
could possibly be useful as an indicate
specimen but further research was re
quired to confirm selenium.

Apnea Direct Injection/Gas
Chromatography
  The apnea sampling method con
bined with direct injection of the expire
air  into a gas chromatograph is an ei
tremely simple and fast method whic
requires no specialized  equipmen
Since no  concentration is achieve!
however, the limit of detection  is reli
lively high for this method.

Breathing Valve/Direct Injec-
tion/Gas Chromatography
  This method, like the apnea/direct it
jection/GC method, is simple and fast.
breathing  valve, air tank, and sampl
collection  bag are required making
slightly more complicated than the a|
nea method. Again, the sample is  ni
concentrated before injection into  tr
gas chromatograph, resulting in red
lively high limits of detection. Becaus
very little sample  manipulation  is pe
formed, recoveries are high. Specific r
covery values were not found in the li
erature reviewed.

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Breathing Valve/Adsorbent
Concentration/Gas Chromatog-
raphy
  This method is complicated and ex-
pensive compared to the direct injection
techniques. The concentration
achieved, however, results in very low
detection limits (as low as 1 part per
trillion). This allows a much larger num-
ber of trace components to be identified
and quantitated, but also leads to more
problems with background interfer-
ences. Some reported recoveries are
high, better than 95%, with coefficients
of variation of 3-6%.
Cryogenic Trapping/Gas Chro-
matography
  This method concentrates  the com-
pounds of interest without concentrat-
ing the oxygen and nitrogen matrix. The
extended sampling period (60 minutes)
results in the equivalent of sub-ppb de-
tection limits although contaminants in
the supply air could increase the limit of
detection for those compounds. The
cryogenic trapping method is sensitive
but also complicated, time-consuming,
and requires a specialized  cryogenic
trapping system. No recovery data are
available for this method.

Tenax GC Cartridge/GC/Mass
Spectrometry
  This method concentrates  the com-
pounds of interest by directly adsorbing
the breath or breath collected in a Ted-
lar bag  onto a Tenax cartridge. The
Tenax cartridge is later thermally de-
sorbed and injected into a  GC/mass
spectrometer. This method has excel-
lent sensitivity but it is costly and re-
quires elaborate equipment.  The per-
cent recovery for laboratory samples
ranges from 77 to 110 percent.

Cascade Impactor Sampling/
Photon Induced X-Ray Emis-
sion  (PIXE) Analysis
  This is a sensitive but very compli-
cated  method for elemental analysis of
breath. Exhaled air (4-6 L) is concen-
trated for PIXE analysis using a cascade
impactor which results in detection lim-
its at the nanogram level. Sample col-
lection efficiency is high (85-99%) which
should lead to high recoveries. The dis-
advantage of this technique  is that a
large amount of specialized and expen-
sive equipment is required  including
cascade impactors,  a computer for
spectral analysis, and a Van de Graff ac-
celerator for PIXE analysis.
Recommendations
  The goal of this research  program
was to evaluate methodologies for de-
termining chemicals of interest to EPA
in human biological matrices. This re-
view included the citations  listed  in
"Chemicals Identified in Human Biolog-
ical Matrices,  A Data Base" and the
Chemical Abstracts  Data Base. Based
on this extensive literature, summary
reviews have been compiled according
to methodology and form the bulk  of
the full report.
  In addition,  based on  the  chemical
and physical characteristics of the com-
pounds of interest and the method by
which they  are determined, some sim-
ple and expected correlations can be
found.
  Some general recommendations can
be summarized as follows:
  1. Many volatile and semivolatile or-
    ganic compounds found in human
    biological  matrices can be deter-
    mined by gas  chromatography
    coupled with a variety of detectors.
    To select  a specific detector, the
    compound type and the matrix
    type being analyzed must be taken
    into account.
  2. Organic compounds of high  boil-
    ing point or that decompose under
    gas chromatographic conditions
    can be determined by liquid chro-
    matography coupled with a variety
    of detectors. Again the selection  of
    a specific detector depends on the
    compound and  matrix type being
    analyzed.
  3. Adsorption onto Tenax cartridges
    is  the  most widely used breath
    sampling  method to preconcen-
    trate organic compounds, includ-
    ing hydrocarbons, chlorinated hy-
    drocarbons,  alcohols, ketones,
    nitrogen-containing compounds
    and others.
  4. Most metals and metal-containing
    compounds can be analyzed by
    atomic absorption spectroscopy or
    inductively coupled argon plasma.
    When the  metals are in solid ma-
    trices such as hair and tissue, they
    can be  determined by neutron acti-
    vation analysis and proton in-
    duced X-ray spectroscopy without
    further sample preparation. Some
    specific metal  matrix combina-
    tions may be best  analyzed by
    electrochemical methods orcolori-
    metric  methods. Other techniques
    applicable to the analysis of metals
    and organometallic compounds
    were found, however, they do not
    appear to be as widespread and
    convenient as the ones mentioned
    above.  With the  exception of
    lithium and beryllium, most metals
    in breath can be determined by
    PIXE analytical techniques.

Body Burden
  Biological monitoring of humans for
xenobiotic body burden entails the col-
lection and analysis of biological speci-
mens (organs, tissues, blood, etc.).
  Biological  monitoring is useful  for
several reasons. First and foremost, it
provides direct evidence of exposure
and absorption of a xenobiotic. It thus
alerts one to the possibility of a human
health hazard. The data obtained by bio-
logical monitoring can be correlated
with the observed level of pollutants in
the environment and with the incidence
of human diseases, which may be due
in part to the presence of the pollutants.
These correlations can be used to deter-
mine  priorities with regard to research
relating to human health and to deter-
mine the necessity of  regulation of pol-
lution sources. Biological monitoring
can point out pollutant problems which
are not otherwise noted, such as could
occur due to bioaccumulation of pollu-
tants  in humans resulting  from  long-
term exposure to very low levels of pol-
lutants in the environment.
  In 1977 an international workshop
was held in Luxembourg on "The Use of
Biological Specimens for the  Assess-
ment  of  Human Exposure to Environ-
mental Pollutants." The objectives  of
the workshop were:
  (a)  To  assess the types of environ-
      mental  pollutants and  human
      specimens most suitable  for bio-
      logical  monitoring and to evalu-
      ate the probable usefulness of bi-
      ological specimen banking.
  (b)  To  examine the state of the art
      and technical feasibility  of pro-
      grams designed to collect human
      biological specimens for  biologi-
      cal  monitoring and  biological
      specimen banking.
  (c)  To  develop guidelines on  sam-
      pling, sample preparation, stor-
      age, and analytical requirements.
  (d)  To  make  recommendations for
      further  research  and develop-
      ment.
  Exposure routes for organic chemi-
cals are  usually inhalation,  ingestion,
and dermal absorption. Once absorbed,
the chemical  may be excreted,  (includ-

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 ing exhalation) stored, or metabolized.
 In general, chemicals are metabolized
 to a more polar  form which leads to
 conjugation and excretion in the urine.
 The fate of any chemical will depend
 upon its volatility, polarity, and chemi-
 cal and biological stability. For example,
 polychlorinated biphenyls (PCB) and
 polychlorinated terphenyls (PCT) are
 nonvolatile, inert, lipophilic compounds
 which  are stored in the adipose tissue.
 More polar compounds such as phenols
 and acids are generally excreted in the
 urine.
  Several  reports found  in  the  pub-
 lished  literature described the simulta-
 neous analysis of many elements in hu-
 man biological  matrices.
  Significant differences in the trace
 levels  of chlorine, potassium, calcium,
 titanium, manganese, iron, copper, and
 lead in hair from different parts of the
 body were found. This suggested that
 the trace elemental absorption and ac-
 cumulation in hair not only depends on
 the particular element, but also on the
 location  of the hair in the body. The
 trace concentrations in white and black
 hair was also found different.
  Seventeen different elements were
 analyzed simultaneously in healthy and
 pathological tissue to obtain informa-
 tion on the cancerous process. Signifi-
 cant differences in the content of vari-
 ous elements were found in the normal
 and pathological tissue. Potassium,
 zinc, and selenium were found in higher
 concentration in the cancerous mucosa
 of the stomach. No significant differ-
 ences  appeared in the elemental com-
 position of blood,  erythrocytes,  hair,
 and  striated muscle taken from the ab-
 dominal wall from patients with gall
 bladder or stomach cancer when com-
 pared with other noncancer diseases.
  Human hair root has been  analyzed
for the presence of several elements.
Hair root, rather than strand may reflect
the most recent exposure influences
and  its analysis is exclusive  of exter-
nally acquired  constituents.  Samples
collected from  23 randomly  selected
rural Florida children, three to six years
of age were analyzed. The results
showed differences in the Fe content by
sex and Cu (and possibly others) differ-
ences caused by intake.
  Human blood serum was analyzed for
 zinc, copper, iron, chromium, man-
 ganese, and selenium as part of a sur-
 vey to determine whether or not the
 Australian aboriginal people received
 optimal diet. Special attention was di-
rected to chromium because of the high
incidence of diabetes mellitus in this
population.
  Multiple-element analysis of human
cerebrospinal fluid  and other tissues
was used to determine diseases of the
nervous system. The  distribution  of
copper, iron, and  zinc  in the  cere-
brospinal fluid was such that deficien-
cies could not be studied. For the major-
ity of trace elements studied, the normal
values could not be determined  much
less the deficiencies. The interpretation
of sporadic high levels was not possi-
ble. High levels of silicon found were
not anticipated. The accumulation of sil-
icon in the body fluids of patients with
renal  failure and on dialysis coupled
with the clinical correlations made sili-
con a candidate for an uremic neuro-
toxin. The higher levels of silicon  in
cerebrospinal  fluid  of clinically-
diagnosed and autopsy-proven cases of
Alzeheimer's disease coupled with the
presence of silicon in the neurofibrillary
tissue of these patients suggest a corre-
lation of silicon levels with this poorly
understood disease.
  Twenty-four elements were  mea-
sured in hair from 20 individuals who
worked in a lead-zinc smelter in Poland.
These same elements were also mea-
sured in hair taken from 20 individuals
considered "normal" or not exposed.
Elements found to be at elevated levels
relative to the controls included As, Se,
Ag, Cd, and Sb.
  L. Sheldon, M. Umafia. J. Bursey, W. Gutknecht, R. Handy,  P. Hyldburg,  L
    Michael, A. Moseley, J. Raymer, D. Smith, C. Sparacino, and M. Warner are
    with Research Triangle Institute, Research Triangle Park, NC 27711
  James Bridges is the EPA Project Officer (see below).
   The Project Summary covers the following reports:
    "Evaluation of Methods for Analysis of Human Fat, Skin Nails, Hair, Blood and
    Urine,"(Order No. PB 85-242 790/AS; Cost: $28.95, subject to change)
    "Methods for Sampling and Analysis  of Breath," (Order No.  PB 85-243
      277'/AS; Cost: $9.95, subject to change).
  The above reports will be available only from:
          National Technical Information Service
          5285 Port Royal Road
          Springfield,  VA 22161
          Telephone: 703-487-4650
  The EPA Project Officer can be contacted at:
          Hazardous Waste Engineering Research Laboratory
          U.S. Environmental Protection Agency
          Cincinnati, OH 45268

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Center for Environmental Research
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