EPA-600/1-76-029
                                                September  1976
ANALYSIS OF BLOOD, HAIR, URINE, AND DUST SAMPLES

              FOR HEAVY METALS
                     By

               Anna M. Yoakum
          Stewart Laboratories, Inc.
            5815 Middlebrook Pike
            Knoxville,  TN  37921
           Contract No. 68-02-2266
               Project Officer

               Dr. Carl Hayes
          Pooulation  Studies  Division
     Health Effects Research Laboratory
     Research Triangle Park, N.C. 27711
                                ...UN AGENCY
    U.S. ENVIRONMENTAL PROTECTION AGENCY
     OFFICE OF RESEARCH AND DEVELOPMENT
     HEALTH EFFECTS RESEARCH LABORATORY
     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.   Approval  does not signify that the contents necessarily
reflect the views and policies  of the U.S. Environmental Protection
Agency, nor does mention of trade names or commercial products
constitute endorsement or recommendation for use.

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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
establishment 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,
environmental 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.

     The chemical analyses provided under this contract support a
collaborative survey by the Center for Disease Control and the Environmental
Protection Agency to assess metal absorption in children living in the
vicinity of primary non-ferrous smelters.  The results of the overall survey
will be reported separately.
                                                H.  Knelson, M.D.
                                               iDirector,
                                     Health  Effects  Research  Laboratory

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                           ABSTRACT

     Communities from ten states in the United States and two
cities in Mexico were studied.   The communities were chosen for
their proximity to primary non-ferrous smelter industries.

     Three lead and five zinc smelter areas were sampled for  blood,
hair, and dust.  Urine, blood,  hair, and dust were collected from
fourteen copper smelter sites and four control cities.

     Samples were analyzed for arsenic, lead, cadmium, copper and
zinc.
                              IV

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                              TABLE OF CONTENTS
                                                                          Page
  I.   Introduction 	  1

 II.   Summary	2

III.   Conclusions  	  4

 IV.   Recommendations	5

  V.   Discussion of Analytical Approach and Results  	  11

      A.  Preliminary Treatment for Sample Preparation 	  11
      B.  Special Research Studies 	  44
      C.  Analytical Methodology and Discussion  	  46
      D.  Review of Quality Control Program	51
      E.  Assessment of Analytical Data	85

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I.   INTRODUCTION







    Documentation of excessive absorption of heavy metals by children




    living near nonferrous metal smelters in the U. S.  A. and Canada




    activated a nationwide study to determine the distribution of several




    heavy metals around primary lead,  zinc,  and copper  smelters.   Pre-




    school children were selected as test subjects because signsof absorp-




    tion of heavy metals are more likely to  appear with them than with




    older children or adults.  The purpose of this project was to analyze




    tissue and dust samples collected  from test subjects living within




    a two mile radius of each smelter  so that heavy metal absorption could




    be evaluated.







    Control sites were selected to provide background reference data as




    to the existing level of these heavy metals in areas not associated




    with nonferrous smelters.

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II.  SUMMARY
     The study sncompassed collections from ten states in the U. S. A.




     and two cities in Mexico.  Tissue and dust samples from preschool




     children and their homes were differentiated for analysis in accord




     with the type smelter or control being evaluated.







     Three lead and five zinc smelter areas were sampled for blood, hair,




     and dust.  Urine, blood, hair, and dust were collected from fourteen




     copper smelter sites and four control cities.







     All dust samples were analyzed for As, Pb, Cd, Cu, and Zn.  Hair




     samples from lead and zinc smelter sites were analyzed for Cd and Pb;




     while copper smelter sites and control areas received As, Pb, and Cd




     determinations.  Urine collections (copper smelter sites and control




     areas) were measured for specific gravity and As.  Lead and FEP were




     analyzed in blood from all three smelter types.  Additionally, Cd




     was included for lead smelter sites;  Cd and Zn for zinc smelter sites;




     and Cu and Zn for copper smelter sites.  Control bloods were tested




     for all five blood parameters.







     Internal and external quality control programs were integrated with




     sample analyses.  Accuracy and precision data derived from these pro-




     grams are reported and analyzed in depth.







     Analyses for each collection site were reported in progressive order




     in accord with a numeric field assignment for individual subjects from

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each participating family.   The laboratory analyst and date for each




determination, including quality controls, was shown within this




format.

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III.   CONCLUSIONS




      The quality of the analytical results produced during this




      study has been evaluated in terms of uncertainty measurements




      which represent at least 95 percent confidence interval based on




      measurement error and variability between samples.   The




      detrimental effects contributed by the samples themselves were




      successfully overcome in all sample types except urine.  Overall




      accuracy and precision,  expressed as relative percent, for each sample




      category is as follows:   dust, + 3%, hair,  + 5%, blood, + 10%,




      and urine, +49%.

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IV.   RECOMMENDATIONS







     Based on the experience gained in the performance of this contract,




     the following recommendations are suggested for the improvement of




     future projects of a similar nature:







     A.   Dust - The smear technique employed in the collection of the dust




         samples has two significant shortcomings.







         A sample of this type does not lend itself to any standard




         analytical quality control program.  Since the entire sample




         cannot be removed, blind sample splits and recovery studies




         cannot be conducted.







         Data obtained can be reported only in terms of micrograms of




         acid soluble metal per towelette  (or per sample).   Since the




         actual amount of sample collected varied widely, the ability




         to express the data as micrograms of metal per gram of dust would




         have allowed for an exact comparison of analytical data between




         collection sites, rather than the relative comparison which results




         from the method employed.







         From a collection standpoint, dust is, perhaps, the most difficult




         sample type addressed in this project.  Paint and other materials




         heavily laden with metals may be  preferentially dissolved by the




         organic liquids in the towelettes.  The abrasive nature of the




         sampling procedure contributes to the incorporation of these con-

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    taminants in the samples, and there is no procedure for obtain-




    ing an initial weight of the dust as it is collected.







    A recommended dust collection technique, which the contractor




    has used, involves dusting a surface of sufficient size to render




    a sample adequate for complete analysis.  A small camel's hair




    brush is used to sweep the sample into a small whirlpak.  This




    disallows external contamination from the hands of field personnel,




    surface finishes, and/or the implement used for collection.  The




    brush must, however, be thoroughly cleaned after each use.







B.  Urine - In order to combat the tendency for urine samples to




    preferentially precipitate with time lapse and temperature varia-




    tions, a change in the collection protocol would be necessary.







    Specific gravity measurements should be performed in the field




    as each sample is collected.







    Since time lapse and temperature variations occur during storage




    and transport to the laboratory, urine samples should be shaken




    and aliquoted in the field as soon after collection as possible.




    If blind splits are to be made, this should also be performed at




    that time.  This procedure would allow for homogeneity of each




    sample, and it would assure uniform analytical handling.







C.  Blood - Since clotted blood does not lend itself to uniform sample




    aliquoting techniques, two avenues are open as approaches to pre-




    clusion of this problem.  First, sample splits could easily be




    made from the syringe while the blood is fresh and not clotted.

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    Secondly, a more uniform mixing with an anticoagulant could be




    applied.  The former would, at this time, appear to be the safest




    and simplest.







    All blood samples should be ejected from the syringe into containers




    appropriate for shipment.  Inverted, leaky syringes held together




    with various types of tape do not constitute proper shipping vessels.







D.  Hair - The extreme variation of metals content in hair indicates




    that small samples are not representative of the overall hair of




    the subject.  Thus, a concerted effort should be made to collect




    as large a sample as possible from each subject.







    Zip-loc bags provide excellent transport containers; however, they




    should be locked carefully as soon as they receive a sample of hair.




    A secure seal allows for transport in a condition isolated from




    external contamination.  Each hair sample should be handled with




    gloves and completely enclosed within the plastic bag.







    Human scalp hair is widely used for assessing environmental ex-




    posure to metals.  Trace metal content of hair is, likewise,




    reported to be an indicator of deficiency conditions in both humans




    and test animals.  Because of these considerations and because of




    the ready availability of hair as a biopsy tissue, scalp hair lends




    itself well to environmental monitoring of humans for trace metal




    exposure.







    A number of problems are apparent which could have significant




    influence on the interpretation of data for hair as a valid

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biological indicator for environmental exposure.  Findings from




a series of pilot research studies conducted by the contractor,




as in-house satellite investigations, clearly indicate the need to




conduct a more closely controlled investigation in the area of




trace metal analysis of hair.   There is, thus, a need for the




design of an analytical study to define, investigate, and establish




the parameters which can adversely influence the interpretation of




hair data.







The recommended research should study, for a period of at least




two years, the trace metal content of scalp hair samples collected




under carefully controlled conditions from a healthy population




living in an area free from major point source atmospheric metal




contamination.  All pertinent background information relating to




the sampling population should be made available so that relation-




ships between trace metal contents and personal covariants can be




evaluated.  This requires an extensive, well-administered question-




naire.  Participation in the program should be limited to those




subjects who are willing and cooperate fully in providing all




pertinent background information to be included in the study;




therefore, the selection of the sampling population should be care-




fully controlled to assure that all vital areas are included in the




study.







Since factors other than environmental exposure can play a signifi-




cant role in trace metal concentrations and distributions, the study




should address itself to the following potential factors:

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    (1)  Seasonal variations—i.e., hair growth rate and




         frequency of hair care.







    (2)  Hair care—i.e., types and frequency of use of




         chemical products and their contribution to




         elemental alteration of  hair.







    (3)  Inherent variabilities—i.e.,  physical and physio-




         logical hair characteristics and metals content along




         single strands of hair.







    (4)  Time effects—i.e., storage time for washed versus




         unwashed hair prior to analysis.







    (5)  Participant characteristics—i.e.,  race, medical




         treatment,  residence location, smoking habits, diet,




         sex, age, etc.






E.  General Sampling Protocol - Consistency in adherence to a sampling




    protocol at each collection site is essential from a quality con-




    trol standpoint.  Certain basic guidelines should be followed in




    order to obtain data which can be compared directly and correlated




    with known varying parameters.  These parameters should have been




    primarily limited to 1) geography,  baseline conditions, and




    environmental exposure characteristics and 2) inherent biological




    uptake of heavy metals.







    In order to produce data which represent a real profile of these




    conditions, one should properly and legibly code and permanently

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label each sample as it is collected.  Each individual sample




should be treated precisely the same as all other samples of




that particular type.  The protocol should be realistic in terms




of maximum time required to perform each field procedure.  For




example, frozen samples shipped over long distances should arrive,




at the laboratory, in the same condition as those shipped from




proximal collection sites.







Provided the recommended protocols for field sample procedures,




as discussed for each sample type, are followed, all collections




could be shipped directly to participating laboratories on an




individual basis.  This would eliminate variables such as time




differentials, thawing of samples, inhomogenous aliquots, and




misplacement of partial batches.







Also, uniformity of sample sizes submitted for analysis should be




held at a reasonable tolerance level.  All containers to be used




in a project of this type should be procured from single line suppliers




and, preferably, from a given manufacturing batch or lot.






Shipment schedules for submission of samples for analysis should be




planned in advance and modes of transport carefully investigated.







A continuous rapport and conscientious use of quality control among




persons responsible for field sampling, laboratory analyses, and




project management are imperative in the obtainment of valid and




useful analytical data.
                           10

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V.  DISCUSSION OF ANALYTICAL APPROACH AND RESULTS







    A.  Preliminary Treatment for Sample Preparation - This project included




        receipt of human tissue and house dust from twenty-four sampling




        sites.  Of these sites, three were near lead smelters, five near




        zinc smelters, twelve near copper smelters, and four were from con-




        trol areas (Table 1).  Because of deviations from the prescribed




        sampling protocol, a laboratory screening of those samples not




        meeting the minimum requirements for analysis was established.  The




        criteria on which samples were rejected are shown in Table 2.   The




        acceptable size limits were prescribed in the contract; however,




        additional measurements were required in the laboratory in order to




        differentiate samples which were very near the limit and could not




        be visually determined.  The various procedures for handling these




        samples will be discussed on an individual basis for each sample




        type, and the rejections made for each collection site will be




        enumerated and related to their cause.







        1.  Procedures for Sample Screening—Each shipment contained




            samples from several collection sites and, therefore, required




            a preliminary sorting and assignment to an organized sequence




            for analysis.  Once this process had been achieved, each




            sample type was individually screened for analytical accept-




            ability.







            a.  Blood—The tubes were checked for cracks and/or probable




                contamination, and notations were recorded when appropriate.




                All samples which were not whole blood or could not be





                                   11

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             TABLE 2.   Criteria for Rejection of Samples
                       from the Project.
Sample Tissue Type

      Blood



      Hair


      Urine



      Dust
   Criteria for Rejection

< 1 ml
not complete whole blood
indeterminant code identification

< 0.2 gm
indeterminant code identification

< 25 ml
indeterminant code identification
samples not contracted

taken on materials other than
towelettes
indeterminant code identification
                               13

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    identified from labels were counted and discarded.




    After thawing, the blood volumes were individually compared




    visually against a reference 1 ml blood sample.  Those




    samples determined to be <1 ml were counted and discarded;




    however, because of excessive clotting and the large number




    of samples which were close to the 1 ml volume, some samples




    were processed and analyzed for FEP and later discarded for




    further analysis.  This was done after the blood was trans-




    ferred to a graduated cylinder for volumetric measurement.




    FEP determinations were made prior to this transfer because




    of the prescribed time requirement in the method.  All




    blood samples which had been rejected were reassembled and




    enumeration checks were finalized.







b.  Hair—Since hair and dust were frequently shipped in bags




    together, a separation and verification of code assign-




    ments of these samples was necessary prior to screening.




    Samples obviously too small and/or with unidentifiable




    labels were rejected.  Improperly sealed bags were noted




    as were containers other than Zip-Iocs.  A second screen-




    ing of hair was attempted by weighing borderline samples




    on a top loading balance.  If the weights were between




    0.20 and 0.25 gm, the samples were reentered and coded for




    analysis.  Some of these samples fell below the 0.2 gm




    limit when reweighed on an analytical balance after they




    had been cleaned and dried.  These final discards were




    added to all of the hair previously rejected, and proper




    notations were recorded.





                        14

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    c>   Urine—Some shipments contained urine from lead and zinc




        smelter sites, which were not to be included for analysis




        under this contract; therefore, these samples were rejected.




        Each of the samples included for analysis was compared,




        after thawing, to a reference urine of a 25 ml volume.  All




        samples failing to meet this criterion were enumerated and




        rejected.







    d.   Dust—After separation from hair samples (see Hair above),




        dust was observed for adherence to sampling protocols and




        analytical preferences.  Since only ten samples per collec-




        tion site were to be chosen, properly sealed bags with no




        evidence of contamination received top consideration.




        Apparent sample size and legibility of label codes were




        then utilized as secondary criteria.  One entire shipment




        of dust from Corpus Christi, Texas, was discarded because




        the samples had been taken on cotton balls and blanks were




        unavailable.  Rejected dust samples were enumerated after




        all these selection criteria had been imposed.






2.   Sample Conditions and Handling Techniques—The sample conditions




    and the various procedures for handling these samples are pre-




    sented on an individual basis for each sample type according




    to  collection site.  A summary of samples received versus samples




    analyzed is given in Table 1.







    a.   Lead Smelters—The lead smelter samples were to be analyzed




        for:  FEP, Pb, and Cd in blood; Pb and Cd in hair; and Pb,
                          15

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Cd, Zn, Cu, and As in dust.  Of the 187 bloods received,




162 were completely analyzed; of the 208 hair samples,




126 were completely analyzed; and of the 158 dust samples,




the prescribed 30 were appropriate for complete analysis.




The following is a resume of the condition of sample ship-




ments and preliminary analytical treatment of samples




received from each lead smelter.
                   16

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                   HERCULANEUM, MO
General - Samples received 9/18/75 with no instructions




          therefore all samples initially coded







  Blood - All thawed; 28 badly clotted therefore duplicate




          FEP's run; condition required special mixing for




          Cd; 11 run for FEP only







   Hair - Zip-Iocs sealed; 36 rejects







  Urine - Received 108 not to be analyzed—all were coded







   Dust - Zip-Iocs not sealed; 77 screened
                      17

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                      BIXBY, MO
General - Samples received 9/25/75







  Blood - Partially thawed; 2 badly clotted therefore




          duplicate FEP's run; condition required special




          mixing for Cd







   Hair - Zip-Iocs and sealed; 39 rejects







  Urine - Samples received but not to be analyzed







   Dust - Zip-Iocs and sealed; 30 screened
                      18

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                     GLOVER, MO
General - Samples received 9/25/75







  Blood - Partially thawed; 1 badly clotted therefore




          duplicate FEP run; condition required special




          mixing for Cd







   Hair - Zip-Iocs and sealed; 7 rejects







  Urine - Samples received but not to be analyzed







   Dust - Zip-Iocs and sealed; 21 screened
                       19

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b.  Zinc Smelters - The zinc smelter samples were to be analyzed




    for FEP, Pb,  Cd, and Zn in blood;  Pb and Cd in hair; and




    Pb, Cd,  Zn, Cu, and As in dust.   Total analyses were com-




    pleted for 370 of the 415 bloods received,  297 of the 445




    hair samples, and 30 of the prescribed 50 from a total of




    241 dust samples.  A resume of each zinc smelter follows.
                         20

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                  BARTLESVILLE, OK
General - Samples received 9/18/75 with no instructions




          therefore all samples initially coded







  Blood - All thawed;  all in inverted syringes; 13 with




          white adhesive tape,  59 with black electrical




          tape, 18 with scotch tape; 3 badly clotted




          therefore duplicate FEP's run; condition




          required special mixing for Cd; 3 for FEP only







   Hair - Alligator sandwich bags with twist ties; 15 rejects







  Urine - Received 75 not to be analyzed—all were coded







   Dust - Alligator sandwich bags with twist ties, towelettes




          very dry; 55 screened
                        21

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                 CORPUS CHRISTI, TX
General - Samples received 11/18/75 and 11/20/75; all




          samples had to be re-coded from information




          received from CDC







  Blood - Partially or completely thawed; received 51




          samples, 38 of which were rejects because they had




          been spun down and only plasma separates were




          shipped; condition required special mixing for




          Cd on the 13 whole blood samples






   Hair - Zip-Iocs sealed; 35 rejects






  Urine - Samples received but not to be analyzed







   Dust - Zip-Iocs sealed but all 46 were rejects because




          they had been taken on cotton balls instead of




          towelettes
                       22

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                     MONACA, PA
General - Samples received 11/18/75, 12/5/75, and 12/16/75







  Blood - First and second shipments partially thawed—third




          shipment (4) not iced at all; 31 badly clotted




          therefore duplicate FEP's run; condition required




          special mixing for Cd; third shipment (4) received




          after coding, causing handling and data processing




          problems because of skips in numerical sequence







   Hair - Zip-Iocs and sealed; 13 rejects







  Urine - Samples received but not to be analyzed







   Dust - No samples received
                       23

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                    PALMERTON, PA












General - Samples received 1/9/76







  Blood - Frozen; 2 badly clotted therefore duplicate




          FEP's run; condition required special mixing




          for Cd







   Hair - Zip-Iocs sealed; 33 rejects







  Urine - Samples received but not to be analyzed






   Dust - Zip-Iocs sealed; 68 screened
                       24

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                    AMARILLO,  TX
General - Samples received 1/9/76







  Blood - Frozen; 1 badly clotted therefore duplicate




          FEP run; condition required special mixing for




          Cd







   Hair - Zip-Iocs mostly sealed; 52 rejects







  Urine - Samples received but not to be analyzed







   Dust - Zip-Iocs mostly sealed; 42 screened
                      25

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c.  Copper Smelters - The copper smelter samples were to be analyzed




    for:  FEP, Pb,  Zn, and Cu in blood; Pb, Cd,  and As in hair;




    specific gravity and As in urine; and Pb, Cd, Zn, Cu, and




    As in dust.  Of the 1046 bloods received, 1026 were completely




    analyzed; of the 1156 hair samples, 951 were analyzed; of the




    977 urines, 793 were analyzed; and of the 812 dust, the pre-




    scribed 120 were appropriate for complete analysis.  A resume




    of each copper smelter follows.
                       26

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                     HAYDEN, AZ
General - Samples received 9/18/75 with no instructions




          therefore all samples initially coded; all samples




          completely mixed with MI







  Blood - Partially thawed; labels extremely difficult




          to read; 12 badly clotted therefore duplicate




          FEP's run







   Hair - Zip-Iocs mostly not sealed; 4 rejects







  Urine - Partially thawed; 8 rejects; precipitated






   Dust - Zip-Iocs mostly not sealed; 59 screened
                       27

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                      MIAMI, AZ
General - Samples received 9/18/75 with no instructions




          therefore all samples initially coded; all samples




          completely mixed with HA







  Blood - Partially thawed; labels difficult to read; 18




          badly clotted therefore duplicate FEP's run







   Hair - Zip-Iocs mostly not sealed; 17 rejects







  Urine - Partially thawed; 9 rejects; precipitated






   Dust - Zip-Iocs mostly not sealed; 62 screened
                      28

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                     MORENCI, AZ












General - Samples received 9/25/75







  Blood - Partially thawed; 7 badly clotted therefore




          duplicate FEP's run







   Hair - Zip-Iocs sealed; 8 rejects







  Urine - Completely thawed; 25 rejects; heavily precipitated







   Dust - Zip-Iocs sealed; 64 screened
                      29

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                       AJO, AZ












General - Samples received 10/15/75 and 10/30/75







  Blood - Partially thawed; 8 badly clotted therefore




          duplicate FEP's run







   Hair - Zip-Iocs sealed; 15 rejects







  Urine - Partially thawed; 9 rejects; precipitated; 6




          samples received too late to run with scheduled




          batch







   Dust - Zip-Iocs sealed; 59 screened
                       30

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                     ANACONDA, MT












General - Samples received 10/30/75







  Blood - Thawed; 3 badly clotted therefore duplicate




          FEP's run







   Hair - Zip-Iocs mostly sealed







  Urine - Partially thawed;  33 rejects; precipitated







   Dust - Zip-Iocs mostly sealed; 47 screened
                        31

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                     McGILL,  NV












General - Samples received 10/30/75







  Blood - Thawed; 4 badly clotted therefore duplicate




          FEP's run







   Hair - Zip-Iocs mostly sealed; 17 rejects







  Urine - Partially thawed; bad leakage in shipping




          container; 4 rejects; precipitated






   Dust - Zip-Iocs mostly sealed; 35 screened
                       32

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                   SAN MANUEL, AZ
General - Samples received 10/30/75







  Blood - Thawed; 14 badly clotted therefore duplicate




          FEP's run; 29 samples sent in inverted syringes,




          untaped; tubes cracked and bad leakage on samples




          and container; codes very difficult to read







   Hair - Zip-Iocs mostly sealed; 97 rejects







  Urine - Thawed; slight leakage in container; 32 rejects;




          heavily precipitated






   Dust - Zip-Iocs mostly sealed; 69 screened
                       33

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                   WHITE PINES, MI












General - Samples received 11/18/75







  Blood - Partially thawed; 1 badly clotted therefore




          duplicate FEP run; 1 FEP only







   Hair - Zip-Iocs partially sealed or not sealed; bags




          contaminated with blood; 5 rejects







  Urine - Partially thawed; codes very difficult to read;




          9 rejects; precipitated






   Dust - Zip-Iocs partially sealed or not sealed; bags




          contaminated with blood; 74 screened
                        34

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                   COPPER HILL,  TN












General - Samples received 11/18/75, 11/20/75, and 12/16/75







  Blood - Partially or completely thawed; 12 badly clotted




          therefore duplicate FEP's run







   Hair - Zip-Iocs sealed; 4 rejects







  Urine - Partially or completely thawed; some codes very




          difficult to read; 19 rejects; heavily precipitated;




          15 samples received too late to run with scheduled




          batch







   Dust - Zip-Iocs sealed; 62 screened
                      35

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                     DOUGLAS, AZ












General - Samples received 11/18/75 and 11/20/75







  Blood - Partially thawed; 7 badly clotted therefore




          duplicate FEP's run







   Hair - Zip-Iocs sealed; 6 rejects







  Urine - Partially thawed; 13 rejects; precipitated







   Dust - Zip-Iocs sealed; 53 screened
                      36

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                 AQUA PRIETA, MEXICO












General - Samples received 11/18/75 and 11/20/75







  Blood - Partially or completely thawed; 11 badly clotted




          therefore duplicate FEP's run







   Hair - Zip-Iocs partially sealed or not sealed; 31




          rejects







  Urine - Partially or completely thawed; 13 rejects;




          heavily precipitated







   Dust - Zip-Iocs partially sealed or not sealed; 44 screened
                      37

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                      HURLEY, NM











General - Samples received 1/9/76






  Blood - Frozen






   Hair - Zip-Iocs sealed; 1 reject






  Urine - Frozen; codes very difficult to read; 19 rejects;




          precipitated






   Dust - Zip-Iocs sealed; 64 screened
                       38

-------
d.  Control Areas - The control samples were to be analyzed




    for:  FEP, Pb, Cd, Zn, and Cu in blood; Pb, Cd, and As




    in hair; specific gravity and As in urine; and Pb, Cd, Zn,




    Cu, and As in dust.  Complete analyses were achieved on




    368 of the 374 bloods, 229 of the 426 hair samples, 291




    of the 373 urines, and the 40 prescribed dust samples from




    a total of 308.  The following is a resume of each control




    area:
                        39

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                     SAFFORD, AZ
General - Samples received 12/5/75 and 12/16/75







  Blood - First shipment frozen or partially thawed; second




          shipment (10) not iced at all; 11 badly clotted




          therefore duplicate FEP's run; condition required




          special mixing for Cd; second shipment (10)




          received after coding, causing handling and data




          processing problems because of skips in numerical




          sequence






   Hair - Zip-Iocs sealed; 56 rejects






  Urine - Partially thawed; 35 rejects; precipitated







   Dust - Zip-Iocs sealed; 65 screened
                      40

-------
                    PERRYVILLE, MO












General - Samples received 12/5/75 and 12/16/75







  Blood - First shipment frozen or partially thawed; second




          shipment (9) not iced at all; 5 badly clotted




          therefore duplicate FEP's run; conditions required




          special mixing for Cd; second shipment (9) received




          after coding, causing handling and data processing




          problems because of skips in numerical sequence







   Hair - Zip-Iocs sealed; 16 rejects







  Urine - Partially thawed; 18 rejects; precipitated






   Dust - Zip-Iocs sealed; 66 screened
                     41

-------
                   ALBUQUERQUE, NM
General - Samples received 12/5/75 and 12/16/75;  all samples




          not properly labeled—lacked child code, sent




          coding sheets to CDC for corrections







  Blood - First shipment frozen or partially thawed; second




          shipment (9) not iced at all; 7 badly clotted




          therefore duplicate FEP's run; condition required




          special mixing for Cd; second shipment  (9) received




          after coding, causing handling and data processing




          problems because of skips in numerical  sequence






   Hair - Zip-Iocs not sealed; 78 rejects







  Urine - Thawed; 22 rejects; heavily precipitated







   Dust - Zip-Iocs not sealed; 68 screened
                      42

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                   NOGALES,  MEXICO
General - Samples received 12/5/75







  Blood - Frozen; 15 badly clotted therefore duplicate




          FEP's run; condition required special mixing for




          Cd







   Hair - Zip-Iocs mostly sealed;  47 rejects







  Urine - Frozen; 7 rejects;  precipitated







   Dust - Zip-Iocs mostly sealed;  69 screened
                     43

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B.  Special Research Studies - The deviations of the actual samples

    received from the Government specification as set forth in the

    RFP were of such magnitude that it was necessary to conduct two

    special research studies before a final analysis scheme could be

    established.


    1.  Urine Precipitate Problems—Essentially, all urine samples

        contained a precipitate.  Since a number of arsenic com-

        pounds are extremely volatile, heating the sample to dis-

        solve the precipitate was not a feasible approach.  The

        best solution to the problem would have been to prepare the

        entire sample, as received, for analysis.  However, this

        solution to the problem was not economically feasible.

        The extreme variability in sample size would have required

        custom adjustments of the amount of digestion acid required

        for dissolution, as well as the final sample volume.  These

        adjustments would, therefore, create preparation procedures

        unique for each individual urine sample.  Additionally, data

        retrieval, key-punching, and verification involvement would

        have increased by more than a factor of three.


        The only practical approach to the problem was to try to

        remove a representative aliquot from each sample.  One of

        the larger urine samples from a lead smelter area was shaken

        vigorously and divided into four aliquots of 25 ml each and

        one final sample of 19 ml.  All were analyzed for arsenic.

        Results:  As  = 34 ppb + 6 ppb  (or 3.4 +  0.6 yg/100 ml)
                  (95% Confidence Interval)

                  Coefficient of Variation = 8.82%
                             44

-------
    Based on these data, this aliquoting approach was adopted

    as a compromise preparation procedure.  Actual experience

    with the samples in the contract revealed that some samples

    produced a relatively homogeneous sample after vigorous shaking

    while others still contained rather large precipitated aggre-

    gates.


2.   Dust Collection Background Study—The RFP specifications

    called  for dust samples of at least 100 mg.   In actuality,

    the dust samples were collected by a technique  which does

    not allow for an actual measurement of the weight of the dust

    collected.  A dust smear was collected from the top of a door

    facing  using a moist, disposable paper towel, size. 14 X 20 cm

    (44 in2) impregnated with 20% denatured alcohol and 1:750

    benzalkonium chloride.  The towelette most frequently used

    was Wash 'n Dri from Canaan Products, Inc.,  Canaan, Conn.

    Because of the inconsistency in the manner by which the samples

    were taken and because of the varying mass of the dust collected,

    the only analysis avenue open was to employ the sample prepara-

    tion procedure used by Vostal and associates in their study.


    Actual  blank towelettes from the lots used to sample the smelters

    were not available for background determinations.  Consequently,

    background data were obtained from one box of 26 towelettes
      Vostal, J. Tares, E. Sayre, J. W.,  and Charney, E.,
      Environmental Health Prospectives,  May, 1974, p 71.
                           45

-------
        supplied to the Contractor by CDC.   The results obtained for




        the towelette background contribution are contained in Table 3.




        Appropriate background corrections  were applied to all dust




        data.   Data obtained by the above described method are




        representative of the 0.1 N HCL soluble content of the samples.




        No relationship can be established  between this value and the




        total metal content of the dust.







C.  Analytical Methodology and Discussion - A number of modifications in




    the analytical approach and methodology proposed by the Contractor




    for the fulfillment of this contract were required because of the




    size and condition of the samples actually received for analysis.




    This section discusses these modifications and the analytical methods




    actually employed.







    1.  FEP Analysis of Blood—The original RFP specified that blood




        samples would be maintained on ice  in the dark from time of




        collection until an aliquot was taken for FEP determination at




        the contracting laboratory.  Based  on the assumption that the




        samples would be handled in the prescribed manner, the method




        chosen by the Contractor for the FEP determination was the




        method of Joselow which measures the fluorescence of zinc




        protoporphyrin.  When the samples arrived, it was learned that




        many had been collected for three months or more and stored in




        a frozen state for varying periods of time.  It was, therefore,




        necessary to change from the method of Joselow to the method of




        Sassa; and measure all erythrocyte porphyrins.  The analytical
                                46

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                TABLE 3.   Towelette Background Data






Concentrations are expressed as total micrograms per towelette
Sample No.
R 3214
R 3215
R 3216
R 3217
R 3218
R 3219
R 3220
R 3221
R 3222
R 3223
Mean Value
Lead
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
<0.25
Cadmium
<0.025
<0.025
<0.025
<0.025
<0.025
<0.025
<0.025
<0.025
<0.025
<0.025
<0.025
Arsenic
<0.025
<0.025
<0.025
<0.025
<0.025
<0.025
<0.025
<0.025
<0.025
<0.025
<0.025
Copper
1.75
1.25
1.63
1.81
1.88
1.40
1.50
1.38
1.63
1.88
1.61
Zinc
3.25
3.00
4.50
1.00
4.25
4.13
11.8
3.50
4.75
3.25
4.34
                               47

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    method of Sassa is the referee method employed by CDC for FEP
                    o
    determinations.
2.   Analysis of Blood for Heavy Metals—The original sample prepara-
    tion proposed by the Contractor called for the samples to be
    lyophilized and for the organic matrix to be destroyed by oxygen-
    flask combustion.  Prior to this step, however, a microsample
    (50 yl) was to be removed for lead analysis by direct flameless
    atomic absorption spectroscopy.  The presence of many macroclotted
    blood samples made it impossible to withdraw valid microsamples
    for the blood lead analyses.  To overcome this problem, the
    proposed sample preparation scheme for blood was replaced with
    a wet, acid-oxidation preparation of the total sample (NIOSH
    Methods P & CAM 101 and 139).

    The acid digest solution was then analyzed by conventional
    atomic absorption for copper and zinc and the lead was deter-
    mined by direct flameless atomic absorption (using a graphite
    furnace with multi-linear temperature programming and simultane-
    ous background correction).

    At this time, a request was received from the Project Officer
    to add an analysis for cadmium on selected blood samples.  Pre-
    liminary investigation revealed that the blank correction for
    the acid-digestion sample preparation was too high for cadmium
    to give reliable analytical data.  The following analytical
    procedure was found to give acceptable results in all cases
    except for those with excessive macroclots.  Blood samples, in
    o
     Granick,  S.,  Sassa,  S.,  Granick,  J.  L. ,  Levere,  R. D., and  Kappas,
     A.  (1972)  Proc.  Nat.  Acad.  Sci.  USA.   69,  9,  2381-2385-

                          Aft

-------
    their original collection containers, were placed on a vortex




    mixer for two minutes.  A 250 yl sample was withdrawn with an




    Eppendorf pipette and diluted to five ml with distilled water.




    An equal aliquot of sample and 1% ammonium sulfate solution




    (Cd free) were placed in a pyrolytic graphite tube and analyzed




    by flameless atomic absorption.







3.  Analysis of Hair for Heavy Metals—All hair samples were washed




    with agitation for 30 minutes in a non-ionic detergent




    (7-X-O-Matic) and thoroughly rinsed with distilled deionized




    water.  Samples were then dried (vacuum oven 60° C, 0.5 atm.)




    and weighed.  The original proposal called for the organic matrix




    to be destroyed by oxygen-flask combustion.  This, however, was




    replaced with the acid-oxidation preparation technique being




    used for blood and urine since the results obtained by both




    techniques are comparable.  Conventional atomic absorption was




    employed for the analysis of cadmium and lead in hair while the




    gaseous hydride atomic absorption method was used for arsenic.







4.  Analysis of Urine for Arsenic—The method originally proposed




    and actually used for the analysis of arsenic in urine is the




    NIOSH Method No. P & CAM 139.  The samples are ashed with a




    mixture of nitric, perchloric, and sulfuric acids to destroy the




    organic matrix.  The ash is treated with ammonium oxalate to




    remove traces of nitric acid and the solution is analyzed by




    atomic absorption of the gaseous hydride (arsine)„







5.  Analysis erf House Dust—The Contractor originally proposed to
                           49

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follow the sample preparation procedure for house dust given




in the RFP as the recommended method.  Specifically, the "as




received" sample was to be weighed and then sieved through a




0.5 mm screen while being shaken for five minutes at 260




oscillations per minute.  The sieved portion was then to be




weighed and extracted with quartz distilled nitric acid (6N)




at 50° C for 30 minutes.  The extract was then to be filtered




and analyzed for Pb, Cd, Zn, and Cu by atomic absorption.







An aliquot of the filtered HNC>3 acid extract was to be treated




to remove all traces of nitric acid and then analyzed for arsenic




by the gaseous hydride-atomic absorption technique.







Additionally, on a selected number of house dust samples, an




extract was to be made on a portion of the unsieved sample and




a comparative analysis was to be made on both sieved and unsieved




portions.






As stated in the previous section of this report, the dust samples




were collected on moist, commercial towelettes.  The sample prepara-




tion method prescribed for use in the RFP is applicable only to




dust free from the collection matrix.  Consequently, the entire




sample, towelette plus dust, was soaked at room temperature for




16 hours in 20 ml of 0.1 N HCL.  The eluates were decanted, final




volume was adjusted to 25 ml, and analyses were performed for the




elements of interest—conventional atomic absorption for Pb, Cd,




Zn, Cu, and atomic absorption of a gaseous hydride for As.






In summary, the employed analytical methodology represents
                        50

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        current, state-or-the-art approaches which are basically sound




        from a technical standpoint.  Documentation of the accuracy and




        precision of the methods are contained in the section of this




        report which deals with the internal analytical quality control




        program.  The limits of detection for the various elements in




        the appropriate analysis solutions are contained in Table 4.







D.  Review of Quality Control Program - This contract required two




    quality control programs—a documented internal analytical quality




    control program, not to exceed 10% of the total effort, and an




    external control program, not to exceed 1%.  All quality control




    efforts were in addition to the contract samples.







    The internal quality control program encompassed blind split sample




    analyses for hair, blood, and urine; blind random analyses of




    standard reference samples; recovery and precision studies from




    two large, composite samples (blood and hair); and the analysis of




    standard sample splits with two reference laboratories.







    The external control program was to consist of blind split sample




    analyses, blind known samples to be analyzed concurrently with




    contract batches of samples, and duplicate determinations to be




    performed by reference laboratories.







    Each 50th blood and urine sample was split into two parts by the




    Contractor and one of the aliquots of each was analyzed by the




    the Contractor.  The other portion was delivered to the EPA Project




    Officer for analysis in EPA1s laboratory.  The Contractor was not




    required to split hair samples for duplicate analysis as a part of




    the Scope of Work of the contract.






                                51

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                  TABLE  4.  Limits of Detection


Concentration units are micrograms per liter (parts per billion)


    Element                                Detection Limit, ppb

Arsenic                                             1.

Copper                                             10.

Zinc                                               10.

Lead
  Conventional A.A.                                10.
  Flameless A.A.                                    0.5

Cadmium
  Conventional A.A.                                 1.
  Flameless A.A.                                    0.02
                                52

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This section of the final report deals with the internal analytical




quality control program.  Because of the importance of laboratory




analyses and the resulting actions which they produce, a program to




insure the reliability of the data is essential.  Data from a valid




analytical quality control program provide an assessment and measure-




ment of the precision and accuracy of the analytical results.  In




addition, a properly designed and conducted program will identify




any segment of the total effort which is "out of control."







1.  Accuracy—Accuracy refers to the degree of difference between




    observed and known, or actual values.  Two approaches were




    used to establish the accuracy of the analytical data—recovery




    studies and the analysis of standard reference materials.




    a.  Recovery Studies—These studies were conducted on an




        actual sample and not on reference standards.  At the




        outset of this project, a pint of blood was secured from




        the local blood bank.  The entire sample was lyophilized




        and homogenized to form a large, stable blood composite




        for use in the internal quality control program.  Known




        amounts of copper, zinc, and lead were added to an aliquot




        of the control blood sample.  Using the composite lyophilized




        blood sample, with known concentrations of 17.5 yg/g zinc,




        4.60 yg/g copper, and 0.79 yg/g lead, recoveries were 98%,




        99%, and 97% respectively.  Individual results of this




        study are found in Table 5.







    b.  Standard Reference Materials (SRM) Analysis—Two standard sam-




        ples were used to provide a measure of the control of accuracy
                             53

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                           TABLE  5.  Recovery Study Data

      Concentration units are micrograms per gram lyophilized whole blood
                                      Copper
Sample No.

R 3007
R 3008
R 3009
R 3010
R 3011
   Sample
Concentration
     4.
     4.
     4.
     4.
 60
 60
 60
 60
     4.60
          Added Spike    Sample Plus  Spike    Percent  Recovery
  ,0
  .0
  ,0
  .0
             3.0
      7.78
      7.35
      7.65
      7.45
      7.36

Average Recovery
102.
 97.
101.
 98.
 97.

 99.%
Sample No.

R 3012
R 3013
R 3014
R 3015
R 3016
    Sample
Concentration
    17.
    17.
    17.
    17.5
    17.5
.5
.5
.5
                                       Zinc
          Added  Spike     Sample Plus  Spike     Percent  Recovery
10.0
10.0
10.0
10.0
10.0
     26.1
     27.0
     28.3
     25.4
     28.0

Average Recovery
 95.
 97.
103.
 92.
103.

 98.%
                                       Lead
Sample No.

R 3017
R 3018
R 3019
R 3020
R 3021
   Sample
Concentration

     0.79
     0.79
     0.79
     0.79
     0.79
           Added  Spike     Sample  Plus  Spike    Percent  Recovery
              1.0
              1.0
              1.0
                   1.68
                   1.88
                   1.70
                   1.64
                   1.73

             Average Recovery
                          94.
                         105.
                          95.
                          92.
                          97.

                          97.%
                                         54

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during the analysis stage of the contract.  These two




samples were Lyophilized Bovine Liver (SRM 1577) from




the National Bureau of Standards and Dried Animal Whole




Blood (Code No. A-2/1974) from the International Atomic




Energy Commission.







Prior to the analysis of any samples from the contract,




eight aliquots from each of these reference materials were




prepared for analysis in order to verify the proposed




analytical methodology.







The bovine liver sample was analyzed first.  Results of




these tests are contained in Table 6 .   These data show




no significant difference between the actual analysis




mean and the certified value for cadmium, lead, and zinc.




However, a relative error of +3.59% was observed in the




case of copper.  Subsequent investigation traced the bias




to the master copper standard employed for the atomic




absorption analysis.  This problem was corrected, and the




animal blood standard samples were analyzed.  Results for




these samples are found in Table  7.  All of the elements




analyzed, including copper, showed no significant difference




between the actual analysis mean and the recommended values




for the standard.







In order to provide a measure of accuracy from analysis




batch to batch, a blind sample of one of these standard




reference samples was analyzed with the samples from each
                     55

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        TABLE 6.   Analysis of Standard Reference Material 1577 Bovine Liver

                  Source:   National Bureau of Standards

                  Concentrations are expressed as micrograms per gram (ppm)
Sample No.            Copper            Zinc            Lead            Cadmium

R 3030                 199.             137.            0.33              0.27
R 3031                 209.             135.            0.25              0.31
R 3032                 201.             131.            0.31              0.28
R 3033                 204.             128.            0.35              0.25
R 3034                 192.             128.            0.35              0.26
R 3035                 188.             133.            0.36              0.29
R 3036                 186.             130.            0.36              0.27
R 3037                 197.             138.            0.36              0.26


Mean Value
  (95% Confidence
   Interval)           197. + 6.6       133. + 3.3      0.33 + 0.03       0.27 + 0.02

Coefficient of
   Variation             4.02%            2.93%        11.4%              7.02%

Standard Values
  (95% Confidence
   Interval)           193. + 10        130. + 10       0.34 + 0.08       0.27 + 0.04
                                         56

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TABLE 7.  Analysis of Dried Animal Whole Blood, Reference Material
          (Code No. A-2/1974)

          Source:  International Atomic Energy Commission

          Concentrations are expressed as micrograms per gram (ppm)
Sample No.

R 3022
R 3023
R 3024
R 3025
R 3026
R 3027
R 3028
R 3029
Copper

 50.5
 51.6
 44.3
 45.0
 51.5
 47.2
 44.1
 43.9
Zinc
88,
90.
89,
86,
88,
90,
91,
88.7
Lead

0.86
0.89
0.99
0.90
0.96
0.90
0.97
0.96
Mean Value
  (95% Confidence
   Interval)

Coefficient of
   Variation

Standard Values
  (95% Confidence
   Interval)
 47.3 + 2.9


  7.26%



 45. + 6
89.2 + 1.2


 1.57%



89. + 9
0.93 + 0.04
5.03%
0.97 + 0.22
                                 57

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        smelter and/or  control  site.   These  data  are  tabulated




        in Tables  8  and 9.   An  examination of  these data  shows




        no significant  deviation from the  standard value  of  the




        reference  samples.







    Based on both  the recovery  studies and the actual analysis of




    these standard reference materials,  it can be stated  that at




    the 95% confidence  interval the overall  accuracy  of  the  analytical




    data is 98% or better.







2.   Precision—Precision is a measure of mutual agreement among




    individual measurements of  the same property under prescribed




    similar conditions.  In an   analytical quality control program,




    precision is determined, not on reference  standards,  but by




    the use of actual samples which incorporate the  inherent matrix




    variables associated with the sample type  under  investigation.







    a.  Quality Control Charts—Analytical quality control charts




        visibly represent the continuing validity of  routine ana-




        lytical data and the performance of  individual analysts




        within a given laboratory.  Limits for these  control charts




        are calculated  using the repeatability standard deviation




        for the control sample.  Upper and lower control limits




         (UCL and LCL) are defined as + 3 standard deviations while




        upper and  lower warning limits (UWL  and LWL)  are set at




        + 2 standard deviations.  For any given period,  if all  points




        on the control chart are within the  control  limits and  some-




        what randomly distributed about the  mean value,  it can be
                            58

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          TABLE 8.  Analysis of IAIA Animal Blood (Code No. A-2/1974)
Sample No.

GF 6129
GF 6131
GF 5079
GF 5080
GF 5861
GF 6040
GF 6240
GF 6385
GF 7164
GF 7363
Batch No.

   2
   2
   3
   3
   4
   4
   5
   5
   6
   6
Copper
   ,5
   .1
   .4
50.
46.
44.
42.8
47.9
51.2
48.
42.
44.6
43.0
   ,7
   .3
Zinc

89.0
94.5
86.3
85.2
88.6
89.0
91.8
93.4
82.9
85.6
Lead

0.86
0.96
0.89
0.90
0.99
0.97
0.90
0.96
0.93
0.99
Mean Value
  (95% Confidence
   Interval)

Coefficient of
   Variation

Standard Values
  (95% Confidence
   Interval)
                     46.2 + 2.3
                      7.06%
                     45. + 6
                 88.6+2.7     0.94+0.03
                  4.23%
                 89. + 9
                               4.87%
                               0.97 + 0.22
                                       59

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               TABLE 9.  Analysis of NBS Bovine Liver (SRM 1577)
Sample No.           Batch No.

GF 5582                 2
GF 5583                 2
GF 5824                 3
GF 5893                 3
GF 5811                 4
GF 5938                 4
GF 6217                 5
GF 6359                 5
GF 7072                 6
GF 7310                 6
GF 7473                 6

Mean Value
  (95% Confidence
   Interval)

Coefficient of
  Variation

Certified Values
  (95% Confidence
   Interval)
Copper

 201.
 194.
 190.
 194.
 189.
 193.
 195.
 201.
 203.
 189.
 194.
 195. + 3.3
   2.51%
 193. + 10
Zinc

135.
128.
133.
130.
136.
128.
128.
130.
126.
134.
131.
  2.51%
130. + 10
 Lead

 0.33
 0.35
 0.25
 0.36
 0.31
 0.36
 0.35
 0.36
 0.30
 0.39
 0.40
131. + 2.2     0.34 + 0.03
12.5%
 0.34 + 0.08
                                       60

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assumed that the precision of the analyses is consistent

with the precision of the internal quality control sample.


The quality control sample used to generate the quality

control charts relating to blood analyses was an aliquot

of the lyophilized blood composite used for the. recovery

study.  A fresh urine composite sample was used for the

arsenic control.  Representative quality control charts

generated through the use of these samples are illustrated

by Figures 1-10.  Typical variability in precision among

analysts is demonstrated by a comparison of Figures 1-3

and 4-6.  These charts profile the precision of the ana-

lytical methodology employed for the analysis of lead,

zinc, and copper in blood and arsenic in urine.  Comparable

charts were also developed for the analysis of arsenic,

lead, and cadmium in hair and cadmium in blood.


A summary of the statistical profile generated by the

control chart data is contained in Table 10.  The following

measures apply:
     Standard Deviation (s)
     the variance.
The square root of
     s =
                     -x2
                  n-l
          where x is the mean
          of the samples.
                    61

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         Coefficient of  Variation (V)  or Percent
         Relative Standard Deviation (% RSD).


                  100 s
                                          where x is the mean and s
             	,  the  standard deviation of
                                          the  sample.
                   x
         95% Confidence Interval.   The interval expressed by

         x + t(v,F=.975)s/^n,  where v = n-1 and where t

         is taken from tables  of the t-distribution,  is

         sufficiently wide to  grant that the mean of  the whole

         population has a 95%  chance of falling  in  the interval.

         Such tables may be found in most of the CRC  mathematics

         publications.


    The average coefficient of variation for the four elements

    is 3.91%.  The range exhibited by V is 1.15% to 5.98%.


b.  Composite Hair Analysis—Two composite hair samples were

    a part of the internal quality control program.  In

    order to establish the inherent variability of a given

    hair sample, a special study was designed using one of

    these composites.  The entire sample (^ 10 grams) was

    washed and dried according to routine standard procedures.

    The sample was then cut into one-quarter inch lengths

    and mixed thoroughly.  Ten aliquots of the dried hair,

    weighing 0.5 gm each, were removed and analyzed individually.

    The remainder of  the hair was initially processed as one

    sample; then, after dissolution, ten aliquots were removed

    from the prepared sample solution and analyzed.
                          62

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                              72

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                        TABLE 10.  Summary of Control Chart Data



                              Copper (ppm)     Zinc (ppm)     Lead  (ppb)     Arsenic  (ppi

Mean                              4.96           17.5           46.8             2.94

Standard Deviation abs.           0.17            0.20           2.80            0.15

Uncertainty Limits
of Mean                   abs. +  0.07          + 0.08         + 1.3          +  0.04
(95% Confidence Interval) rel %+  1.31          +0.43         +2.69         +1.24

Coefficient of Variation          3.44%           1.15%          5.98%           5.06%
                                          73

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        A comparison of  the  precision measurements between the




        4.49 gm dissolved  composite (Table 11 )  and the ten




        individual 0.5 gm  dry hair aliquots  (Table 12) clearly




        differentiates the reproducibility of the analytical




        instrumentation, as  defined by the former, from the




        inherent elemental variability in hair, as shown by the




        latter.   These data  conclusively demonstrate the signifi-




        cance and magnitude  of this variation in trace metal




        concentrations when  small individual hair samples are




        compared analytically.  A comparative summary of these




        statistical profiles is found in Table 13.







        Another study was  conducted on a second hair composite.




        The composite was  divided into thirteen individual 0.5 gm




        samples.  These  samples were submitted to the laboratory




        as blind samples in  the last three batches of the project.




        The analytical results are given in  Table 14.






        Based on these  three studies, the innate variability of




        certain elements in  hair was clearly demonstrated.  This




        fact will have a significant bearing on the accurate




        interpretation of  the blind sample  split analyses con-




        ducted for this  contract.  This topic will be discussed




        in detail in a  later section of this report.







3.  Interlaboratory Comparison Analyses







    a. Comparative Arsenic Analyses—Concurrent with this contract




       effort, the Contractor, was employed  by EPA to analyze

-------
           TABLE 11.  Analytical Data from 10 Aliquots of Composite Hair Sample
                      (4.49 grams) - Prepared as One Single Sample.


                      Concentration units are micrograms per gram
Sample No.

BB 007-1
BB 007-2
BB 007-3
BB 007-4
BB 007-5
BB 007-6
BB 007-7
BB 007-8
BB 007-9
BB 007-10
Lead

10.0
10.0
10.0
 9.85
 9.80
 9.90
10.0
10.1
10.0
10.0
Cadmium
    45
    50
    45
    45
    45
    45
    45
    45
    50
  1.45
Arsenic

 <0.05
 <0.05
 <0.05
 <0.05
 <0.05
 <0.05
 <0.05
 <0.05
 <0.05
 <0.05
Copper
 24,
 24.
 24.4
 24.6
 24.
 24,
 24,
 24,
 24.6
 24.3
.3
.6
.3
.3
,3
.3
Zinc

497.
497.
497.
497.
497.
497.
500.
494.
494.
494.
Mean Value  abs.
(95% Confidence
  Interval)
 9.97 + 0.06
Standard Deviation,
  abs.               0.09
  1.46 + 0.02
                0.02
 <0.05
 24.4 + 0.10
                                    0.14
             496.  ^
Coefficient of
  Variation
 0.90%
  1.44%
                   0.58%
                  0.3*
                                           75

-------
                  TABLE 12.  Analytical Data for Ten Individual 0.5 gram

                             Aliquots of Composite Hair Sample

                             Concentration units are micrograms per gram
Sample No.
Z 019
Z 020
Z 021
Z 022
Z 023
Z 024
Z 025
Z 026
  027
  028
Lead

 8.80
10.0
11.3
12.5
10.0
10.0
10.0
17.5
11.3
10.0
Cadmium

 0.67
 0.67
 1.11
 1.22
 0.67
 0.75
 1.50
 2.13
 1.38
 0.88
Arsenic

 <0.05
 <0.05
  0.10
 <0.05
 <0.05
  0.10
  0.06
 <0.05
 <0.05
 <0.05
Copper
 24.9
   .1
   .1
   .1
30.
27.
27,
24.6
25.9
27.1
28.6
25.1
29.1
Zinc

506.
593.
521.
516.
481.
491.
526.
518.
488.
538.
Mean Value abs.
(95% Confidence
 Interval)
11.1 + 1.76
Standard Deviation,
  abs.               2.46
Coefficient of
  Variation
22.1%
 1.10 + 0.34
                0.48
43.4%
 <0.06
 27.0 + 1.34   518. + 22.9
                                   1.87
                   6.95%
                                 32.
                 6.19%
                                             76

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                     TABLE 13.  Summary-Hair Study Precision Data
Mean Value, 95%
Confidence Interval,
  Composite
  Individual
                               Lead
 9.97 + 0.06
11.1  + 1.76
Standard Deviation
  Composite                 0.09
  Individual                2.46

Coefficient of Variation
  Composite                 0.90%
  Individual               22.1%
                   Cadmium
1.46 + 0.02
1.10 + 0.34
                 0.02
                 0.48
                 1.44%
                43.4%
                    Copper
24.4 + 0.10
27.0 + 1.34
                   0.14
                   1.87
                   0.58%
                   6.95%
                 Zinc
496 + 1.35
518 + 22.9
                 1.90
                32.
                 0.38%
                 6.19%
                                          77

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            TABLE  14.   Analysis of  Quality Control Hair Composite No. 2

                       Concentration units are micrograms per gram
 ample  No.            Batch No.

 F  5264                 4
 T  5377                 4
 tf1  5487                 4
 IF  5578                 4
 IF  6727                 5
JF  6774                 5
iF  6846                 5
 IF  6920                 5
 JF  8270                 6
JF  8347                 6
;F  8389                 6
iF  8482                 6
^F  8561                 6

lean Value abs.
(95% Confidence
 Interval)

standard Deviation,  abs.

 loefficient of Variation
 Lead

 5.00
12.5
11.3
13.8
11.3
12.5
11.3
15.0
10.0
12.5
11.3
10.0
 7.50
11.1 + 1.57

 2.6

23.5%
Cadmium

 <0.25
  0.38
  0.25
  0.25
  0.25
  0.38
  0.25
  0.25
  0.75
  0.38
  0.75
  0.25
 <0.25
  0.36 + 0.11

  0.18

 51.3%
Arsenic

 0.25
 0.075
 0.35
 N. A.*
 0.40
 0.33
 0.28
 N. A.*
 0.28
 0.28
 N. A.*
 N. A.*
 0.25
 0.09

32.7%
*N.  A. = Not Analyzed.  Sample was controlling a zinc smelter
                                        78

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    arsenic in particulate matter.   The analytical methodology




    employed in this effort was NIOSH Method P & CAM 139 (arsenic




    in urine and air)  which is the  identical method used for




    Contract No. 68-02-2266.  In the initial phase of the




    aforementioned project, a correlation study was conducted




    to compare the NIOSH arsenic method with EPA flameless




    atomic absorption technique. Results of these analyses on




    sample splits provided and analyzed by EPA are given in




    Table 15.  The correlation coefficient for these data is 0.9943.







b.  Comparative Blood Lead Analyses—On December 30, 1975, the




    Contractor received twelve reference blood samples from




    the Center for Disease Control  (CDC).  The Contractor ana-




    lyzed these samples by the graphite furnace atomic absorption




    technique.  In addition, the three samples with the highest




    lead values were also analyzed  by conventional atomic absorp-




    tion.







    A comparison of the data obtained by the Contractor with the




    CDC standard values is shown in Table 16.  Performance




    evaluation by CDC was based on  the following criteria:




         For values ^ 40 yg/100 ml, + 15%




         For values < 40 yg/100 ml, + 6 yg




    In only one case out of twelve, did the Contractor's data




    differ from the CDC value by more than + 6 yg.  Sample 5-27




    differed by 6.2 yg.







c.  Interlaboratory Study Conclusions—Based on the results of
                         79

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TABLE 15.   Comparison of Analytical Results of the NIOSH and EPA Arsenic Methods


           Concentrations are micrograms Arsenic per milliliter



                           Contractor Results             EPA Results
      Sample No.        (NIOSH Method P & CAM 139)       (Flameless AA)

         4                       2.00                        1.7

         7                      20.0                        19.2

         12                     15.A                        19.5

         13                      1.56                        1.6

         14                      5.60                        4.8

         15                      6.10                        6.5

         20                     44.0                        44.8
                                      80

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                   TABLE 16•   Comparative Data - Blood Lead
         (Concentrations are expressed as ug Pb/100 ml whole blood)
Contractor Code
      CDC Code
               Contractor Result
               CDC Value
GF 6958
GF 6958*
GF 6959
GF 6960
GF 6961
GF 6962
GF 6963
GF 6964
GF 6965
GF 6965*
GF 6966
GF 6967
GF 6968
GF 6969
GF 6969*
5-29
5-29 Direct Aspiration
5-18
5-36
5-15
5-12
5-27
5-24
5-35
5-35 Direct Aspiration
5-33
5-32
5-30
5-28
5-28 Direct Aspiration
76.6 + 3.6
76.4 + 2.1
12.1 + 2.1
25.5 + 2.0
13.4 + 1.0
22.7 + 0.8
38.0 + 0.9
37.2 + 1.3
55.4 + 1.0
49.1 + 1.5
40.8 + 0.8
56.1 + 0.7
44.9 + 1.0
78.3 + 1.8
81.3 + 2.0
71.8 + 4.6

14.4 + 1.9
26.4 + 2.8
16.3 + 2.2
19.7 + 1.7
31.8 + 1.1
33.2 + 1.7
50.2 + 2.6

37.2 + 2.4
59.4 + 3.6
41.5 + 2.6
79.0 + 3.1

*Direct Aspiration Atomic Absorption Data
A sample of NBS bovine liver was also analyzed with these samples.
are expressed as yg Pb/gram of SRM.
                                               These results
Contractor Code
GF 6970
NBS Code
 SRM 1577
Contractor Result
   0.33 + 0.04
NBS Certified Value
   0.34 + 0.08
                                    81

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        these two interlaboratory comparisions between the Contractor




        and the reference laboratories of EPA and CDC, it can be




        concluded that the accuracy data generated by the internal




        quality control program of the Contractor provide a valid




        assessment of the quality of the analytical data produced




        under this contract.







4.   Blind Sample Split Analyses—A large portion of the internal




    analytical quality control program encompassed blind split




    sample analysis of hair,  blood, and urine.  A measure of the




    statistical profile of these data is the coefficient of varia-




    tion for the split sample pairs.  Complete data for this phase




    of the contract effort are appended.  Comparative precision




    data for the appropriate type of composite control sample and




    the mean value for the actual sample splits are contained in




    Table  17.  If all pertinent criteria for effective quality con-




    trol have been satisfied, the Coefficient of Variation should




    be approximately equal for the control samples and the actual




    sample splits.






    An analysis of the data immediately indicates that some phase




    of the program is "out of control."  The final segment of this




    report section is devoted to an assessment of the internal




    analytical quality control program and related ramifications.







5.   Assessment of Analytical Quality Control Program—A quality




    control program in the laboratory has two primary functions.




    First, the program should monitor the reliability of the results
                             82

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   TABLE 17.  Coefficient of Variation Comparison for Control
              Samples and Mean of Actual Sample Splits.
                                                V for Mean of Actual
                  V for Control Samples            Sample Splits	

Hair
  Lead                    5.98%                       15.2%
  Cadmium                 6.15%                       17.5%
  Arsenic                 5.06%                       14.8%

Blood
  Lead                    5.98%                       22.0%
  Copper                  3.44%                       10.6%
  Zinc                    1.55%                        9.0%
  FEP                  Not Determined                  4.51%

Urine
  Arsenic                 5.06%                       24.9%
                              83

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reported.  It should continually provide an answer to, "How




good are the submitted results?"  This phase may be termed




"measurement of quality."  The second function is the control




of quality in order to meet the program requirements for reli-




ability.







The control of analytical performance in the laboratory is




based on the assumption that a "valid sample" has been submitted




for analysis.  A "valid sample" implies that the sample was




properly taken, preserved, and delivered to the laboratory in




a condition appropriate for all analytical techniques to be




employed.  Further, the validity of controlled analytical per-




formance is dependent upon the use of currently recognized




analytical methods substantiated by the recording and reporting




of subsequent laboratory results in a systematic, uniform, and




permanent fashion.  It must be recognized, however, that quality




control begins with the sample collection and does not end until




the resulting data are reported.  The laboratory control of




analytical performance is but one essential link in obtaining




reliable data.  Each single phase of a quality control effort




will only be as good as the poorest, least controlled area which




has an effect on the results.







A review of  the internal analytical quality control program




precision, accuracy, and interlaboratory comparison data on




standard samples revealed that all phases of the analytical per-




formance in  the laboratory were "under control."  Based on the




data from the composite hair study and visual observations of
                         84

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        the blood and urine samples as they were received in the laboratory,




        the "out of control" portion of the project was readily identifi-




        able as the samples themselves.  Analytical quality control is




        based on the assumption that a homogeneous sample, free from




        obvious external contamination, has been analyzed.  All urine




        samples received for analysis contained varying amounts of pre-




        cipitate, and many of the blood samples contained macroclots.




        Hair samples have an inherent variability and cannot be considered




        homogeneous tissue samples.  In addition, the hair samples were




        extremely small; and most split analyses were performed on samples




        sizes of "j 0.25 gin.







        When all pertinent factors are considered, it can be concluded




        that the Coefficients of Variation obtained for the blind sample




        splits represent sample inhomogeneity rather than the nonreproduc-




        ibility of precision for the analytical methodology.  This con-




        clusion is in no way detrimental to the internal analytical




        quality control program.  It does, in fact, demonstrate that the




        program was properly designed and conducted since the "out of




        control" fraction was isolated and identified.






E.  Assessment of Analytical Data - The problem of less than ideal samples




    was readily apparent as soon as the first shipment of samples arrived




    in the laboratory.  Appropriate method modification to minimize the




    overall effect on the project were suggested by the Contractor and




    approved by the Project Officer.  These modifications were designed to




    overcome sample shortcomings in all areas except those which were
                                85

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exclusively dependent on sample homogeneity—specifically, blind




sample split precision.   An overall assessment of the analytical




data for each sample type will now be presented.






1.  Hair—With the exception of the samples used in the blind




    split quality control program, the total hair sample as received




    was analyzed.  The uncertainty of the data obtained from those




    samples which were analyzed in their entirety (>90%) and not




    split for quality control, correlates directly to the accuracy




    and precision of the analytical method as previously presented.




    The mean value for the blind sample split analyses is the most




    accurate representation for these samples.  This mean value has




    a comparable accuracy to the samples which were analyzed as a




    whole.






2.  Blood—The situation for blood is identical to that for hair.






3.  Dust—All dust samples were analyzed as total sample received.




    On an absolute basis, the data are as good as the accuracy and




    precision of the methods employed .






4.  Urine—The urine analyses are the least precise data in the entire




    study.  Because of an extreme variation in sample size, it was




    not analytically feasible to perform the analyses on the total




    sample as received.  The nature and amount of precipitate changed




    drastically from sample to sample.  Every effort was made to




    remove a representative aliquot for analysis; however, the magni-




    tude of the Coefficient of Variation for the blind sample splits
                            86

-------
(24.9%) indicates that the difficulty was not overcome in many




cases.  While the urine data are less precise on an absolute




basis, it should be pointed out that on a relative basis the data




are still meaningful for collection site comparisons.   For




example, the median value for arsenic in urine for site AJ is




9.77 yg/100 ml, for site HA, 3.06 yg/100 ml, and for site PV,




0.49 yg/100 ml.  Thus it is apparent that the lack of precision




in the urine data definitely does not preclude assessment of




the relative significance of the analytical results for the




study.  It should, however, be pointed out that the validity




of specific gravity measurements taken on precipitated samples




has not been established.







The measures of accuracy formulated in section D of this report




can be applied directly to an evaluation of all data for a given




element analyzed in a specific matrix.  A summary treatment of the




analytical means of each type of determination for the four matrices




from various collection sites is shown in Tables 18 through  21.




The accuracy of these values, and the individual data from which




they were generated, is basically equivalent to that achieved




analytically on standard reference materials.







In the past, it has been customary to exclude "less-than" values




from chemical analytical data before performing statistical ana-




lysis.  This is regrettable as the less-than values may reflect




the condition of an important subset of the data, and their




omission possibly precludes the likelihood that the finished
                         87

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              TABLE 18.   Analytical Data Summary - Hair
             Concentration units are micrograms per gram

        "Range indicated is the mean + 2 standard deviations"
Collection Site
   Lead
  Number of
  Positive Results
  Cadmium
  Arsenic
AJ
AL
AM
AN
AP
BV
BX
CC
CH
DO
GL
HA
HK
HL
MG
MI
MN
MO
NG
PL
PV
SA
SM
WP
All Sites
19.2 + 0.3
17.5 + 0.3
38.4 + 0.7
25.4 + 0.5
23.5 + 0.4
56.7 + 1.
170. + 3.
45.8 + 0.8
22.8 + 0.4
26.7 + 0.5
51.1 + 0.9
26.4 + 0.5
64.0+1.
20.4 + 0.4
15.9 + 0.3
31.7 + 0.6
20.4 + 0.4
17.6 + 0.3
15.9 + 0.3
44.4 + 0.8
12.6 + 0.2
11.9 + 0.2
19.3 + 0.3
13.9 + 0.3
29.3 + 0.5
2.68 + 0.08
0.69 + 0.02
4.11 + 0.12
1.94 + 0.06
2.22 + 0.06
9.14 + 0.26
7.94 + 0.23
2.78 + 0.08
2.80 + 0.08
1.88 + 0.05
6.35 + 0.18
1.81 + 0.05
4.26 + 0.12
1.65 + 0.05
1.33 + 0.04
1.90 + 0.06
2.13 + 0.06
1.96 + 0.06
1.29 + 0.04
7.40 + 0.21
1.18 + 0.03
1.09 + 0.03
1.26 + 0.04
1.84 ± 0.05
2.95 + 0.08
1.91 + 0.19
0.10 + 0.01

20.3 + 2.1
0.48 + 0.05



0.14 + 0.02
0.43 + 0.04

2.09 + 0.21

0.18 + 0.02
0.37 + 0.04
0.47 + 0.05

0.31 + 0.03
0.41 + 0.04

0.09 + 0.01
0.10 + 0.01
0.23 + 0.02
0.43 + 0.04
2.14 + 0.22
1577
1451
1126
                                  88

-------
     TABLE 19.   Analytical Data Summary - Blood
      Concentrations are yg/100 ml whole blood
"Range indicated is the mean + 2 standard deviations"
Collection
Site
AJ
AL
AM
AN
AP
BV
BX
CC
CH
DO
GL
HA
HK
HL
MG
MI
MN
MO
NG
PL
PV
SA
SM
WP
All Sites
Number of
Positive
Results
22.
26.
28.
26.
35.
26.
33.
28.
18.
30.
22.
30.
26.
34.
19.
27.
25.
23.
37.
35.
21.
23.
23.
17.
27.
FEP
1 +
1 +
8 +
8 +
8 +
5 +
9 +
6 +
5 +
0 +
2 +
4 +
8 +
1 +
4 +
3 +
9 +
8 +
0 +
6 +
0 +
1 +
8 +
3 +
1 +
Lead
1.7
2.0
2.2
2.1
2.8
2.1
2.6
2.2
1.4
2.3
1.7
2.4
2.1
2.7
1.5
2.1
2.0
1.8
2.9
2.7
1.6
1.8
1.9
1.3
2.1
1941
12.5
17.7
22.7
13.4
19.6
28.9
13.7
19.6
16.5
20.5
12.6
21.2
18.8
17.1
+ 2.0
+ 2.8
+ 3.6
+ 2.1
+ 3.1
+ 4.6
+ 2.2
+ 3.1
+ 2.6
+ 3.3
+ 2.0
+ 3.4
+ 3.0
± 2-7

0
0


0
0
0


0

0

Cadmium

.21 + 0.03
.12 + 0.02


.51 + 0.07
.12 + 0.02
.16 + 0.02


.16 + 0.02

.32 + 0.05

9.1 + 1.5
17.3
14.8
13.9
15.3
17.9
16.9
15.2
18.0
18.6
17.5
1921
+ 2.8
+ 2.4
+ 2.2
+ 2.4
+ 2.9
+ 2.7
+ 2.4
+ 2.9
+ 3.0
+ 2.8

0

0
0
0
0


0

.22 + 0.03

.29 + 0.04
.37 + 0.05
.24 + 0.03
.18 + 0.03


.26 + 0.04
844
Co]
105.
99.4

90.3
95.7



97.7
94.6

152.

75.5
96.6
126.

106.
104.

92.7
78.1
106.
106.
103.
1392
Pper Zinc
± 7.
± 7-

+ 6 .
+ 6 .



~t" 6 .
+ 6.

+ 10.

+ 5.
+ 6.
± 8-

± 7.
± 7.

+ 6.
+ 5.
+ 7.
+ 7.
± 7-

372.
368.
371.
372.
320.
552.

295.
346.
339.

521.

383.
352.
424.
351.
340.
352.
362.
372.
346.
362.
374.
379.
1762
± 18
± 18
T J_O
"T" J-O
± 16
± 27

± 15
+ 17
± 17

+ 26

± 19
± 17
± 21
± 17
± 17
+ 17
± 18
+ 18
± 17
± 18
± 18
± 19

                         89

-------
      TABLE 20.   Analytical  Data Summary - Dust

 Concentrations  are absolute micrograms per towelette
"Range indicated is the mean + 2 standard deviations"
allection
Site
AJ
AL
AM
AN
AP
BV
BX
CC
CH
DO
GL
HA
HK
HL
MG
MI
MN
MO
NG
PL
PV
SA
SM
WP
U.1 Sites
lumber of
Positive
Results
Lead
44.2 + 0.8
17.7 + 0.4
35.9 + 0.6
45.2 + 0.8
48.4 + 0.9
76.2 + 1.4
293. + 5.3

82.5 + 1.5
142. + 2.6
37.5 + 0.7
56.4 + 1.0
81.0+1.5
44.1 + 0.8
17.4 + 0.3
19.6 + 0.4

22.2 + 0.4
37.4 + 0.7
86.1 + 1.6
25.2 + 0.5
24.1 + 0.4
25.5 + 0.5
50.2 + 0.9
59.8 + 1.1
220
Cadmium
1.73 + 0.05
0.88 + 0.03
1.50 + 0.04
2.22 + 0.06
1.34 + 0.04
8.64 + 0.25
0.49 + 0.01

1.74 + 0.05
2.86 + 0.08
0.71 + 0.02
2.35 + 0.07
1.18 + 0.03
0.61 + 0.02
0.82 + 0.02
0.85 + 0.02

1.51 + 0.04
0.92 + 0.03
10.3 + 0.30
1.23 + 0.04
1.14 + 0.03
0.42 + 0.01
1.51 + 0.04
2.10 + 0.06
220
Arsenic
3.97 + 0.4
0.09 + 0.009
0.50 + 0.05
34.2 + 3.4
1.18 + 0.12
0.77 + 0.08
0.27 + 0.03

0.40 + 0.04
3.67 + 0.37
0.09 + 0.009
8.93 + 0.9
0.12 + 0.01
0.27 + 0.03
0.91 + 0.09
0.56 + 0.06

0.41 + 0.04
1.43 + 0.14
2.35 + 0.24
0.15 + 0.02
0.31 + 0.03
0.41 + 0.04
0.44 + 0.04
2.87 + 0.29
214
Copper
128. +1.5
559. + 6.
423. + 5.
122. + 1.4
123 + 1.4
17.6 + 0.2
2.14 + 0.03

78.1 + 0.9
462. + 5.4
123. + 1.4
709. + 8.
1023 + 12.
408. +5.
56.9 + 0.7
107. +1.2

158. + 1.8
63.2 + 0.7
140. + 1.6
2.73 + 0.03
21.3 + 0.25
35.3 + 0.41
1784. + 21.
295. + 3.
220
Zinc
260. + 2.
96.6 + 0.7
241. + 1.8
111. + 0.8
89.7 + 0.7
546. +4.
114. + 0.9

149. + 1.1
192. + 1.5
43.8 + 0.33
471. + 4.
179. + 1.4
50.2 + 0.38
39.3 + 0.30
110. + 0.83

275. + 2.
98.3 + 0.7
1616. + 12.
60.0 + 0.5
145. + 1.1
40.7 + 0.3
434. +3.
247. +2.
219
                          90

-------
                     TABLE 21.   Analytical  Data  Summary -  Urine

                       Concentrations  are micrograms  As/100 ml
                                                     Uncertainty*  Calculated  Using
Collection
Sites
AJ
AL
AN
AP
CH
DO
HA
HL
MG
MI
MO
NG
PV
SA
SM
WP
All Sites
Mean Coefficient
of Variation
10.8%
30.6%
12.3%
24.7%
10.5%
38.2%
16.5%
33.5%
22.8%
15.4%
3.9%
16.9%
61.9%
43.5%
27.7%
20.7%
24.9%
Mean
Value
11.53
1.33
3.82
2.19
2.19
2.75
3.81
1.19
2.41
2.53
2.28
2.30
1.34
1.90
2.80
2.50
3.07
Actual Site V
+ 2.5
+ 0.8
+ 0.9
± 1-1
+ 0.5
± 2-1
± 1-3
+ 0.5
± 1-1
+ 0.8
+ 0.2
+ 0.8
± 1-7
± 1-7
+ 1.6
+ 1.0

Mean V for Project
+ 5.7
+ 0.7
+ 1-9
± 1-1
+ 1.1
± !-4
± 1-9
+ 0.6
+ 1.2
± !-3
± l-1
+ 1.1
+ 0.7
+ 0.9
± I-*
± I-2
+ 1.5
Number of
Positive Results
1034
    Uncertainty measurements indicate tolerance limits of + two standard
     deviations.
                                         91

-------
statistics are representative of the population from which the




samples were taken.  A technique exists for working  with data




having less-thans all of the same value (i.e. truncated data),




but no technique has been established for analyzing data having




variable less-thans.







If all less-thans are considered to be in fact zero, a mean taken




from the data is a low bound on all possible means that may be




taken from the data by any technique.  Similarly, if the less—




thans are considered positive at the value stated for the




less-than value, a high bound on the mean may be set.  The mean




best fitting the data must lie between these two extremes.







The data in Tables  18-22  have been treated in the customary




manner.  A comparison of the means determined from consideration




of only positive results and the means determined by the high




and low bounds on these groups of data in which less-than values




appeared is contained in Table 22.  For most of the data from




this study, the calculation of a mean from only positive results




produces very little alteration of the population representation.
                         92

-------
TABLE 22.   Mean Ranges of Groups Containing Less-than Values

Collection
Site
AJ
AJ
AL
AL
AL
AL
AM
AM
AN
AP
AP
AP
AP
BV
BV
BX
CC
CH
CH
CH
DO
DO
DO
GL
GL
GL
GL
HA
HA
HK
HL
HL
HL
MG
MG

Sample
Type
Hair
Hair
Dust
Hair
Hair
Urine
Blood
Hair
Hair
Hair
Hair
Hair
Urine
Blood
Blood
Dust
Hair
Hair
Hair
Urine
Hair
Hair
Hair
Blood
Dust
Dust
Hair
Hair
Urine
Dust
Hair
Hair
Urine
Hair
Hair


Element
Cd
Pb
As
As
Cd
As
Cd
Cd
Cd
As
Cd
Pb
As
Cd
Pb
As
Cd
As
Cd
As
As
Cd
Pb
Pb
As
Zn
Pb
As
As
As
As
Cd
As
As
Cd
Mean
of
Positives
2.684
19.185
0.090
0.096
0.690
1.328
0.120
4.118
1.943
0.477
2.224
23.450
2.187
0.508
28.909
0.271
2.779
0.144
2.803
2.185
0.430
1.876
26.739
12.559
0.094
43.817
51.161
2.086
3.812
0.117
0.183
1.647
1.193
0.369
1.330
Mean
High
Bound
2.660
18.850
0.077
0.089
0.486
1.231
0.117
3.562
1.868
0.467
2.030
23.202
2.131
0.502
28.624
0.222
2.536
0.139
2.718
2.057
0.426
1.733
25.985
12.091
0.087
39.443
49.636
2.066
3.723
0.108
0.174
1.474
1.135
0.357
0.860
Mean
Low
Bound
2.653
18.739
0.072
0.083
0.390
1.223
0.115
3.501
1.854
0.464
1.983
23.133
2.129
0.501
28.577
0.217
2.486
0.133
2.712
2.051
0.425
1.699
25.895
12.013
0.084
39.436
49.456
2.066
3.721
0.105
0.169
1.437
1.130
0.354
0.679
                              93

-------
TABLE 22.   Mean Ranges of Groups Containing Less-than Values
           (con't)
Collection
Site
MG
MI
MN
MO
NG
NG
NG
NG
PL
PV
PV
PV
PV
SA
SA
SA
SA
SM
SM
SM
WP
WP
WP
WP
Sample
Type
Hair
Urine
Blood
Urine
Hair
Hair
Hair
Urine
Hair
Dust
Hair
Hair
Urine
Hair
Hair
Hair
Urine
Hair
Hair
Hair
Hair
Hair
Hair
Urine

Element
Pb
As
Cd
As
As
Cd
Pb
As
Cd
As
As
Cd
As
As
Cd
Pb
As
As
Cd
Pb
As
Cd
Pb
As
Mean
of
Positives
15.888
2.531
0.223
2.281
0.410
1.289
15.922
2.301
7.389
0.154
0.092
1.176
1.340
0.100
0.092
11.861
1.902
0.228
1.262
19.318
0.428
1.844
13.907
2.502
Mean
High
Bound
14.570
2.463
0.220
2.252
0.391
0.878
15.393
2.208
7.294
0.128
0.082
1.049
1.049
0.100
0.972
11.672
1.728
0.189
0.990
15.182
0.419
1.574
13.348
2.462
Mean
Low
Bound
14.197
2.461
0.220
2.251
0.385
0.664
15.209
2.204
7.289
0.123
0.075
1.026
1.025
0.096
0.945
11.633
1.718
0.163
0.811
13.799
0.417
1.525
13.220
2.460
                                 94

-------
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TECHNICAL REPORT DATA
fl'luiisc read In^lritctwm on the reverse before completing)
1 REPORT NO |2
EPA-600/1 -76-029 1
4. TITLE AND SUBTITLE
ANALYSIS OF BLOOD, HAIR, URINE, AND DUST SAMPLES
FOR HEAVY METALS
7 AUTHOR(S)
Anna M. Yoakum
9 PERFORMING ORGANIZATION NAME AND ADDRESS
Stewart Laboratories, Inc.
5815 Middlebrook Pike
Knoxville, TN 37921
12, SPONSORING AGENCY NAME AND ADDRESS
Health Effects Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Rpqparrh THanglp Park, N r P7711
15. SUPPLEMENTARY NOTES
3. RECIPIENT'S ACCESSI ON" NO.
5 REPORT DATE
September 1976
6. PERFORMING ORGANIZATION CODE
8. PERFORMING ORGANIZATION REPORT NO.
10. PROGRAM ELEMENT NO.
1AA601
11. CONTRACT/GRANT NO.
68-02-2266
13. TYPE OF REPORT AND PERIOD COVERED
14. SPONSORING AGENCY CODE
EPA-ORD

16. ABSTRACT
Communities from ten states in the United States and two cities in Mexico
were studied. The communities were chosen for their proximity to primary
non-ferrous smelter industries.
Three lead and five zinc smelter areas were sampled for blood, hair, and
dust. Urine, blood, hair, and dust were collected from fourteen copper smelter
sites and four control cities.
Samples were analyzed for arsenic, lead, cadmium, copper and zinc.
17. KEY WORDS AND DOCUMENT ANALYSIS
a. DESCRIPTORS b.lDENTIFI
arsenic sampling
lead Ulood
cadmium .
-r IE?
smelters
13 DISTRIBUTION STATEMENT 19 SECURI
DTI TAT TO PIIPI TP UNLL
KLLLMoL 1U rUbLIU 2o SECURI
UNCL
ERS/OPEN ENDED TERMS C. COSATI Field/Group
06, A, F
TY CLASS (This, Report/ 21 NO. OF PAGES
ASSIFIED 12°
TY CLASS (Thispage) 22 PRICE
ASSIFIED
EPA Form 2220-1 (9-73)
                                                         106

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