United States
                Environmental Protection
                Agency
               Environmental Criteria and
               Assessment Office
               Research Triangle Park NC 27711
EPA-600/8-83-014A f
July 1983       **
External Review Draft
                Research and Development
r/EPA
Health Assessment
Document for
Chromium
 Review
 Draft
 (Do Not
 Cite or Quote)
                               NOTICE

                This document is a preliminary draft. It has not been formally
                released by EPA and should not at this stage be construed to
                represent Agency policy. It is being circulated for comment on its
                technical accuracy and policy implications.

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                                        EPA-600/8-83-014A
                                        July 1983
                                        External Review Draft
     Health  Assessment
Document for Chromium
                  NOTICE

   This  document is a preliminary draft.  It
   has not been formally released by EPA and
   should not at this stage be construed to
   represent Agency policy.  It is being
   circulated for comment on its technical
   accuracy and policy implications.
       U.S. Environment Protf~tion
       Rc-^c.i V. '.'•••:•.-: •/
       ?/:,J So.h': L      •-' Street
       Chicago, Illinois  60604
      U.S. ENVIRONMENTAL PROTECTION AGENCY
         Office of Research and Development
     Office of Health and Environmental Assessment
      Environmental Criteria and Assessment Office
         Research Triangle Park, NC 2771 1
                  July 1983

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                                      DISCLAIMER








         The report  is  an external  draft for  review  purposes only  and does  not



    constitute Agency Policy.  Mention of trade names  or commercial products does not



    constitute endorsement or  recommendation for use.
U,S. Environmental Protectfcrt "Agency
                                          ii

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                                    PREFACE








     The Office of Health and Environmental Assessment has prepared this health




assessment to serve as a "source document" for EPA use.  This health assessment



document  was developed  for use  by  the  Office  of Air  Quality  Planning and



Standards to support decision-making  regarding  possible  regulation of chromium




as a hazardous air pollutant.



     In  the  development of the assessment  document,  the scientific literature



has been  inventoried,  key studies have been  evaluated and summary/conclusions



have been prepared so  that  chemical's toxicity  and related characteristics are



qualitatively  identified.   Observed  effect  levels  and  other  measures  of



dose-response relationships are discussed, where appropriate,  so  that the nature



of  the  adverse  health  response  are placed   in  perspective  with  observed



environmental levels.
                                      iii

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     The Office  of  Health and Environmental Assessment  (OHEA)  of U.S. EPA is

responsible   for  the  preparation   of  this   health   assessment   document.

Dr. Si Duk Lee,  as  the  Project  Coordinator,  had overall  responsibility for

coordination and direction of the document, preparation and production  effort.

The chapters addressing physical and chemical properties, sampling and analysis,

air quality and toxicity  data were  prepared  by  Life and  Environmental Sciences,

Syracuse Research Corporation, Syracuse,  NY.   The principal  authors of  these

chapters are listed below.



        Dr. Dipak Basu
        Karen Blackburn
        Dr. Bruce Harris
        Dr. Michael W. Neal
        Frederick W. Stoss

     The OHEA Carcinogen  Assessment Group  (CAG) was responsible  for  preparation

of the sections on carcinogenicity.  Participating members of the CAG are listed

below  (principal  authors  of present  carcinogenicity materials are  designated

by •).


        Roy Albert,  M.D.  (Chairman)
        Elizabeth L. Anderson,  Ph.D.
        Larry D. Anderson, Ph.D.
        Steven Bayard, Ph.D.
        David L. Bayliss,  M.S.
        Chao W. Chen,  Ph.D.
       •Herman J. Gibbs,  M.S.,  M.P.H.
        Bernard H,  Haberman,  D.V.M., M.S.
       •Charralingayya B.  Hiremath, Ph.D.
        Robert McGaughy,  Ph.D.
       *Debdas Mukerjee,  Ph.D.
        Dharm V. Singh, D.V.M., Ph.D.
        Todd W. Thorslund, Sc.D.
                                      iv

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    The following individuals  provided peer-review of this draft or earlier-
drafts of this document:

    U.S. Environmental  Protection Agency

    Joseph Padgett
    Office of Air Quality Planning and Standards
    U.S. EPA

    Karen Blanchard
    Office of Air Quality Planning and Standards
    U.S. EPA

    Mike Waters,  Ph.D.
    Office of Health Research
    Health Effects Research  Laboratory
    U.S. EPA

    Jerry F. Stara, D.V.M.
    Office of Health and  Environmental  Assessment
    Environmental  Criteria  and Assessment Office
    U.S. EPA

    Lester D. Grant, Ph.D.
    Office of Health and  Environmental  Assessment
    Environmental  Criteria  and Assessment Office
    U.S. EPA

    Carol  Sakai,  Ph.D.
    Office of Health and  Environmental  Assessment
    Reproductive  Effects  Assessment Group
    U.S. EPA

    External  Peer Reviewers

    Dr.  Ann Baetjer
    John Hopkins  School of  Hygiene
    615  North Wolfe Street
    Baltimore,  MD   21218

    Dr.  Derek Hodgson
    Professor,  Chemistry  Department
    University of North Carolina
    Chapel  Hill,  NC  27514

    Dr.  Marshall  Johnson
    Thomas  Jefferson Medical College
    Anatomy Department
    1020 Locust Street
    Philadelphia,  PA 19107

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Dr. Magnus Piscator
University of Pittsburg
Graduate School of Public Health
Environmental Epidemiology
Pittsburgh, PA  15261

Dr. Bruce Stuart
Stauffer Chemical Company
Farmington Avenue
Farmington, CT  06032

Dr. Robert Tardiff
1*423 Trapline Court
Viena, VA  22180

Dr. Norman M. Trieff
University of Texas Medical Branch
Department of Pathology, UTMB
Gavleston, TX  77550

Dr. Jim Withey
Health Protection Branch
Dept. National Health and Welfare
Tunney's Pasture
Ottawa, Ontario
CANADA K1A 012
                                 vi

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                               TABLE OF CONTENTS

                                                                           Page

DISCLAIMER		    ii

PREFACE 	   iii

LIST OF AUTHORS 	    iv

TABLE OF CONTENTS 	   vii

LIST OF TABLES 	    xi

LIST OF FIGURES 	   xiv

1.   INTRODUCTION 	   1-1

2.   SUMMARY AND CONCLUSIONS 	   2-1

     2.1  BACKGROUND INFORMATION 	            2-1
     2.2  ANALYSIS OF CHROMIUM	   2-2
     2.3  BIOLOGICAL SIGNIFICANCE AND ADVERSE HEALTH EFFECTS
          OF CHROMIUM	   2-3

        2.3-1   Chromium Pharmacokinetics 	   2-3
        2.3.2   Subcellular and Cellular Aspects of
                Chromium Toxicity 	   2-4
        2.3.3   Systemic Toxicity of Chromium	   2-5
        2.3.4   Chromium Carcinogenesis 	   2-5
        2.3-5   Dermatological Aspects of Chromium 	   2-6
        2.3.6   Chromium as an Essential Element 	   2-7

     2.4  HUMAN BIOLOGICAL MONITORING 	   2-7

        2.4.1   Chromium in Blood	   2-7
        2.4.2   Chromium in Urine 	   2-8
        2.4.3   Chromium in Human Hair 	   2-9

     2.5  HUMAN HEALTH RISK ASSESSMENT OF CHROMIUM 	   2-9

        2.5.1   Health Effects Summary	   2-9
        2.5.2   Populations at Risk 	  2-10

3.    BACKGROUND INFORMATION 	   3-1

     3.1  CHEMICAL AND PHYSICAL PROPERTIES 	   3-1
                                      vii

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                           TABLE  OF  CONTENTS  (cont.)
     3.2  PRODUCTION,  USE,  AND RELEASES TO THE ENVIRONMENT 	   3-6

        3.2.1    Production  of Chromium Compounds	   3-6
        3.2.2    Uses of Chromium and Its Compounds	   3-6
        3.2.3    Releases to the Environment	  3-12

     3.3  ENVIRONMENTAL FATE AND TRANSPORT	  3-15

        3.3.1    Air		  3-15
        3.3.2    Water and Sediments 	  3-17
        3.3.3    Soil			  3-19

     3.4  LEVELS OF CHROMIUM IN VARIOUS MEDIA	  3-20

        3.4.1    Ambient Air	  3-20
        3.4.2    Aquatic Media			  3-24
        3.4.3    Aquatic Suspended Materials and Suspended	  3-27
        3.4.4    Soil	  3-29
        3.4.5    Food 	  3-29
        3.4.6    Cigarettes	  3-34

     3.5  INDICES OF EXPOSURE AND DOSE-RESPONSE RELATIONSHIPS	  3-34

        3.5.1    Chromium in Blood 	  3-35
        3.5.2    Chromium in Urine	  3-37
        3.5.3    Chromium in Human Hair	  3-39

     3.6  SUMMARY	  3-40

4.   SAMPLING  AND ANALYSIS"	   4-1

     4.1  SAMPLING AND STORAGE 	   4-1

        4.1.1    Air	   4-1
        4.1.2    Water	   4-3
        4.1.3    Soil and Sediments	   4-4
        4.1.4    Food	   4-5
        4.1.5    Biological  Samples	   4-5

     4.2  SAMPLE PRETREATMENT	   4-5

        4.2.1    Wet and Dry Ashing 	   4-6
        4.2.2    Precipitation	   4-6
        4.2.3    Solvent Extraction	   4-7
        4.2.4    Chromatographic Method	   4-7
                                     viii

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                           TABLE OF CONTENTS (cont.)
     4.3  METHODS OF ANALYSIS 	   4-8

        4.3.1   Atomic Absorption Spectrometry (flame) 	  4-12
        4.3-2   Atomic Absorption Spectrometry (flameless)	  4-12
        4.3.3   Emission Spectroscopy 	  4-14
        4.3.4   Neutron Activation Analysis 	  4-15
        4.3.5   X-ray Fluorescence 	  4-16
        4.3.6   Colorimetric 	  4-1?
        4.3.7   Gas Chromatography 	  4-17
        4.3.8   Chemiluminescence	  4-18
        4.3.9   Polarography 	  4-19
        4.3.10  Mass Spectrometry	  4-19
        4.3.11  Catalytic Method 	  4-19
        4.3.12  Liquid Chromatography 	  4-20

     4.4  CONSIDERATIONS IN ANALYSIS 	  4-20
     4.5  COMPARISON OF METHODS	  4-22

5.   CHROMIUM METABOLISM IN MAN AND ANIMALS	   5-1

     5.1  ROUTES OF CHROMIUM ABSORPTION 	   5-1

        5.1.1   Chromium Absorption and Deposition by Inhalation 	   5-1
        5.1.2   Gastrointestinal Absorption of Chromium 	   5-7
        5.1.3   Chromium Absorption Through the Skin	   5-9

     5.2  CHROMIUM TRANSPORT, METABOLISM,  DISTRIBUTION,
          AND ELIMINATION 	  5-11

        5.2.1   Transport and Metabolism 	  5-11
        5.2.2   Distribution 	  5-14
        5.2.3   Elimination 	  5-19

     5.3  SUMMARY 	  5-22

6.   CHROMIUM AS AN ESSENTIAL ELEMENT 	   6-1

     6.1  CHROMIUM DEFICIENCY 	   6-1
     6.2  GLUCOSE TOLERANCE FACTOR	   6-4
     6.3  SUMMARY 	   6-6
                                      ix

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                           TABLE OF CONTENTS (cont.)

                                                                           Page

7.   CHROMIUM TOXICOLOGY 	   7-1

     7.1  ACUTE EFFECTS OF CHROMIUM EXPOSURE IN MAN AND ANIMALS 	   7-1

        7.1.1   Human Studies	   7-1
        7.1.2   Animal Studies 	   7-1

     7.2  CHRONIC EFFECTS OF CHROMIUM EXPOSURE IN MAN AND ANIMALS 	   7-6

        7.2.1   Evaluation of the Carcinogenicity of Chromium 	   7-6
        7.2.2   Genotoxicity 	  7-86
        7.2.3   Developmental Toxicity and Other Reproductive Effects ... 7-106
        7.2.*J   Chromium Hypersensitivity 	 7-112
        7.2.5   Other Toxic Effects of Chromium	 7-119

     7.3  SUMMARY OF TOXIC EFFECTS OTHER THAN CANCER FOLLOWING
          EXPOSURE TO CHROMIUM COMPOUNDS 	 7-140

8.   CURRENT REGULATIONS AND STANDARDS	   8-1

     8.1  OCCUPATIONAL EXPOSURE	   8-1
     8.2  EXPOSURE TO CHROMIUM IN AMBIENT WATER	   8-1
     8.3  EXPOSURE TO CHROMIUM IN AMBIENT AIR	   8-5

9.   REFERENCES	.-'...		   9-1

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                                LIST OF TABLES

Table                                                                      Page

3-1     Physical Properties of Selected Trivalent Chromium Compounds 	   3-3

3-2     Physical Properties of Selected Hexavalent Chromium Compounds ....   3-4

3-3     Manufacturers and Their Production Capacities of Sodium
        Chromate and Sodium Bichromate 	   3-8

3-4     Principal United States Manufacturers of Chromic Acid 	   3-9

3-5     United States Chromium Consumption Pattern in 1979 	  3-10

3-6     Sources and Estimates of United States Atmospheric Chromium
        Emissions in 1970	3-14

3-7     Regional Distribution of Principal Chromium Emissions 	  3-16

3-8     Five Forms of Chromium Transported in the Yukon and
        Amazon Rivers	  3-18

3-9     Total Chromium Concentrations Measured in the Ambient Air of
        Selected Sites in the United States During 1977-1980 	  3-21

3-10    Chromium Levels in a Few Surface Waters and Groundwaters	  3-25

3-11    Chromium Concentrations in U.S. Drinking Waters 	  3-26

3-12    Concentration of Chromium in Sediments	  3-28

3-13    Chromium Content in Selected in United States' Soils	3-30

3-14    Chromium Content in Various U.S. Foods 	  3-31

3-15    Concentration of Chromium in a Few Commerical Grade Acidic
        Foods 	  3-33

4-1     Analytical Methods for the Determination of Chromium	   4-9

6-1     Estimated Adequate and Safe Intake (EASI) for Chromium 	   6-2

7-1     Inhalation Exposure of Mice to Chromium-Containing Dust 	   7-8

7-2     Carcinomas Produced with Chromium Compounds in Rats 	  7-13

7-3     Lung Tumors Found and Microscopically Confirmed	  7-15

7-4     Exposure Schedule for Bioassay of Chromium Compounds by
        Intrapleural Injection 	  7-18
                                      xi

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                                LIST OF TABLES

Table                                                                      Page


7-5     Compounds Reported to Have Been Tested for  Carcinogenicity
        by Intrapleural Implantation	   7-20

7-6     Experimental Conditions Used to Study the Effect  of Intrafemoral,
        Intraperitoneal, and Intravenous Administration of  Chromium 	   7-23

7-7     Levels of Hexavalent Chromium in Fractionated  Residue  Dust	   7-2H

7-8     Compounds Reported to Have Been Tested for  Carcinogenicity
        by Intramuscular Implantation	   7-27

7-9     Carcinogenicity of Chromium Compounds in Experimental  Animals  ...   7-30

7-10    Location of Chromate Manufacturing Plants Which Participated
        in Epidemiologic Studies and Plants from Which Vital Statistics
        Were Obtained for Each Study	   7-37

7-11    Observed Number of Deaths, Standardized Mortality Ratios  (SRMs),
        and 95$ Confidence Limits (95%  CL) for Deaths  Due to Cancer of the
        Trachea, Bronchus, and Lung and the Number  of  Reported Deaths
        for Which No Certificate Could  Be Obtained,  By Year of Initial
        Employment, Exposure Category and Total Duration  Employed,  For
        Workers Initially Hired as Hourly Employees 	   7-^6

7-12    Lung Cancer in Workers in the Chromate Pigment Industry	   7-58

7-13    Age-Specific Lung Cancer Death  and the Gradient Exposure  to
        Total Chromium	   7-71

7-1*1    Combined Age-Specific Lung Cancer Death Rates  and Total
        Chromium Exposure (in ng/m )	     7-73

7-15    Relative Carcinogenic Potencies Among 53 Chemicals  Evaluated by
        the Carcinogen Assessment Group as Suspect  Human  Carcinogens ....   7-79

7-16    Number of Cigarettes Smoked Per Day	   7-83

7-17    The In Vitro Mutagenicity Bioassay of Chronic  Compounds  	   7-87

7-18    Chromium Produced Clastogenic Effects and Cell Transformation  ...   7-99

7-19    Teratogenic and Fetotoxic Effects of Chromium	7-109

7-20    Studies Suggesting NOAELS or NOELS	7-121
                                     xii

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                            LIST OF TABLES (cont.)

Table

7-21    Clinical Findings in Workers Employed in Chromium-Plating
        Plants 	7-125

7-22    Perforation of Nasal Septum in Chromate Workers 	  7-127

7-23    Perforation of Nasal Septum in Chromate Workers 	  7-129

7-24    Nasal Medical Findings in a Chromium-Plating Plant 	  7-130

7-25    Medical Complaints of Workers in "hard" Chromium Electroplating
        Plant 	  7-137

8-1     Recommended Occupational Standards and Recommended Criteria for
        Chromium Compounds in the United States 	  8-2

8-2     Recommended Standards for Chromium in Ambient Waters in the
        United States 	  8-3

8-3     Ambient Water Quality Criteria for the Protection of
        Human Health 	  8-i*

8-U     Calculated Ambient Water Quality Criteria for the Protection
        of Aquatic Life 	  8-6
                                     xiii

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                        LIST OF FIGURES
Figure
3-1
5-1
5-2
7-1

Simplified Flow Chart for the Production of Metallic Chromium
Rate of blood clearance of intravenously injected Cr(III)
51
Whole-body elimination of intravenously administered Cr(III)
Histogram representing frequency distribution of the potency
Page
3-7
5-15
5-21

indices of 52 suspect carcinogens evaluated by the Carcinogen
Assessment Group	   7-78
                              xiv

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                                1.  INTRODUCTION
     The 1970 Clean Air Act and its 1977 amendment mandate EPA to regulate, under
Section 112,  those  pollutants that "may reasonably be anticipated to result in an
increase in mortality or an increase in serious irreversible,  or incapacitating
reversible, illness."  It also states that EPA must regulate,  under Section 111
(d), those pollutants  that "may reasonably  be anticipated to  endanger  public
health or welfare."
     For  this  reason,  the  Office of  Air Quality  Planning and Standards  has
requested that the Environmental Criteria  and Assessment Office prepare a scien-
tific assessment for  chromium  so that it can be determined whether the regulation
would be warranted under these sections of the Clean Air Act,  since human expo-
sure to  chromium  has been a matter of public  health  concern.   Therefore,  this
chromium document will serve as a scientific  data  base  for regulatory decision
making by the agency. The health assessment document should represent an inter-
pretive summary of relevant studies  rather than a compendium of  all  available
papers.
     The  present  document represents a comprehensive data  base that  considers
all sources of chromium in the  environment, the  likelihood for  its exposure to
humans, and the possible  consequences to man and lower organisms  from its absorp-
tion.  This information is integrated into a format that can serve as the basis
for qualitative and quantitative risk assessments, while at  the same time identi-
fying gaps in our knowledge that limit accurate health assessment at this time.
Thus, it is expected that this document may serve the information needs of other
government agencies  and  the private sector  that may  be  involved  in  decision
making and regulatory activities.
                                      1-1

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                          2.   SUMMARY AND CONCLUSIONS








2.1. BACKGROUND INFORMATION








     Chromium is  a  metallic  element which,  when found in nature,  is  a stable




mixture of four separate isotopes.  Inorganic chromium compounds occur in valence




states ranging from -2 to +6;  however,  in the environment the Cr(III) and Cr(VI)




states are the most stable. Chemically, the Cr(III) state is the most stable and




important form of inorganic chromium.  Cr(VI) compounds  are the most commercially




important  form  of chromium,  and they  also appear to  be  the most  significant




chromium compounds from an environmental standpoint.   Because Cr(VI) is readily




reduced in the presence of organic material, it is rarely found in nature apart




from deposition by anthropogenic sources.



     Although chromite ore is  not  currently mined in the United States, several




chromium chemicals are domestically  produced from imported  ores.   Sodium chro-




mate,  sodium dichromate,  and  chromic acid are three of  the more  important




commercial chromium compounds produced in the United  States.  Metallurgical uses




constitute about  60% of  the  largest  market demand for  chromium.  Chemical uses




are the second largest consumption  sector at 21$,  followed by refractory uses at




18$.



     Chromium emissions are released into the air, water,  and land environments




from a variety of industrial  source  categories.   The largest chromium emission




sources to  the  air  are ore -refining plants, coal combustion units, refractory




plants, and  steel and alloy  plants.   Principal  sources  of chromium  in water




systems include electroplating operations, leather tanneries, and textile manu-




facturing operations.  Significant sources  of  chromium-containing solid wastes
                                      2-1

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 that  are land disposed  include  chromite ore  refining operations and  chromium

 chemical production plants.


      Recent  monitoring of the ambient air  in  many urban and rural area  of  the

 country  has  shown 24-hour average chromium  concentrations  to  be  in  the  range of


 0.0052  to  0.1568 (ig/m3.  The  maximum concentration  determined  during any  one

                                o
 measurement  was  about 2.U8 ng/nr.   The  chromium  concentration  in U.S.  waters


 varies with  the type of surrounding  industrial  sources and the type of  underlying


 soils.   An  analysis  of 383^ tap  waters in  representative  U.S.  cities  showed  a


 chromium concentration ranging from O.1* to 8 ppb.  Chromium levels in  soils vary


 with  soil  origin and  the  degree  of contamination from anthropogenic  chromium


 sources.  Tests on domestic  soils have shown chromium concentrations ranging from

 1 to  1000 ppm, with the average concentration  ranging from  1H to  about  70 ppm.




 2.2. ANALYSIS OF CHROMIUM




     Based on versatility, sensitivity, and precision, the  three  most important


 methods  that have found wide application  for  the analysis  of  chromium   are


 graphite furnace AAS, (atomic absorption spectrophotometry) x-ray fluorescence,


 and neutron  activation analysis.  Of  these  three methods,  graphite furnace  AAS

 has the  advantage of  being the  least expensive method.   The  disadvantage of

 graphite furnace AAS  is  that  it  cannot be  used  for simultaneous multi-element

 analysis.  X-ray fluorescence  has  the advantage over both neutron  activation  and


 graphite furnace AAS analysis in that  it  can  differentiate between the various


 oxidation states of  Cr  without prior pretreatment  of  the  samples.   The use of

 neutron activation analysis normally requires  about a 2-week  cooling period if


post-irradiation separations  are  not  performed.    Thus,  the  technique is   not

suited for  on-line or rapid  analysis of chromium.
                                      2-2

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     The choice of a particular analytical  method  for Cr analysis is dictated by




several factors including the  type of sample to be analyzed, concentration of Cr




in the sample,  and the  scope of  the analysis.  These factors, in combination with



others such  as the desired precision  and  accuracy and  the cost  of analysis,




should be weighed in selecting a particular analytical method.








2.3.  BIOLOGICAL SIGNIFICANCE AND ADVERSE HEALTH EFFECTS OF CHROMIUM








2.3.1.  Chromium Pharmacokinetics.  There is little specific information on the




deposition and adsorption of chromium by the lungs.  As with other particles and




aerosols, deposition will depend on  particle size  and aerodynamic diameter.  The




deposition  of  particles  larger  than  1  |im will  be governed  by  gravitational




settling and impact.  Deposition of  these large particles will occur in the upper



portion of the respiratory tract.  Deposition of smaller particles, £0.5 |im, is




predominantly  governed by diffusion,  and  occurs  in  the  deep  portions of the




lungs.  A portion of the  chromium absorbed following inhalation may result from




mucocilliary  clearance • of particles   in  the  upper  airways with  subsequent




swallowing and gastrointestinal  adsorption.   It has been shown in experimental




animals, however, that the. efficiency of gastrointestinal  adsorption  is low and




of  the order of  <5%.   All  aspects of  chromium  metabolism  are  confounded by



limitations  in current understanding of the  role of valence state and  chemical




form on interaction within biological systems.  Chromium may be absorbed via the




skin,  lungs, or  gastrointestinal tract. Data relating to pulmonary  absorption



are  extremely  fragmented, and  quantification of  absorption is  not possible.




Dermal absorption has  been better studied  in  that factors  affecting the process




are more clearly delineated.  Dermal absorption appears to  be affected  by valence




state,  the  particular  salt  employed,  concentration  applied,  and  pH.   Gastro
                                      2-3

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intestinal absorption is reported  to  be poor  in  all instances;  however,  data




relating  to  factors  affecting gastrointestinal  absorption are  incomplete  and




contradictory. Transport occurs primarily via plasma siderophilin, while Cr(VI)



penetrates red blood  cells.




     Reduction of  Cr(VI)   to  Cr(III)  appears  to occur  rapidly  in  biological




systems, while the  mechanism and kinetics are not completely understood.  Oxida-




tion of Cr(III) from  tissue depots has been proposed, but not demonstrated.




     Chromium, when given in excess amounts, accumulates with time.  Lung, liver,




spleen, kidney, and bone marrow appear to be primary sites of deposition.  Excre-




tion is primarily via the urine, and a small amount possibly excreted through the




gastrointestinal tract.  Distribution and elimination kinetics appear to conform




to a three compartment model.








2.3.2.  Subcellular and Cellular Aspects of  Chromium Toxicity.   Cr(VI) readily




crosses  cell  membranes.    While  early  data  indicate that  cell  membranes  are




impermeable to Cr(III),  later  studies indicate  that  there  is some transport,  but




to a much smaller extent than  with the hexavalent salts.   Intracellularly, only




Cr(III) has been identified.   Recent data suggest that Cr(IIl) may be the  intra-




cellularly reactive species in mutagenesis.  Acute studies indicate target organ




toxicity  results  from cellular  necrosis  once  intracellular chromium reaches




critical  levels.   Effects  of  excess  chromium  on  cellular metabolism  following




either acute or chronic studies has not been investigated thoroughly.




     The mutagenicity of chromium compounds has been examined in a wide variety




of in vitro assays  such as  the reverse and forward mutation, gene conversion, and




DNA modification tests.  In general,  soluble Cr(VI) compounds  are less active




when metabolic activating systems are present and more active in their absence.




Insoluble Cr(VI) and  Cr(III)  compounds are  generally  less active  in  cellular
                                      2-1

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mutagenicity assays.   The  reduction  of Cr(VI)  to Cr(III) by  cellular agents




present in metabolic activation systems may  explain  Cr(VI)'s  lack of mutagenic




activity.  More recent evidence implicating  both Cr(VI)  and Cr(III) in induced



mutagenesis has been reported  in DNA  interaction and DNA polymerase infidelity




assays.








2.3.3.  Systemic Toxicity of Chromium.








     2.3.3.1.   ANIMAL  DATA — Ingestion of  soluble  Cr(Vl)  solutions can cause




local irritation  but,  generally,  chromium salts  are  relatively non-toxic when




administered  orally.    Acute  exposures   (intraperitoneal)  result in kidney




failure,  liver, heart, and  brain micropathology.  Chronic target organ effects




have  not  been described.   Inhalation exposures  result  in characteristic lung




alterations, such  as  congestion,  granuloma, and  thickening of  alveolar walls;




adverse  effects  in other organs following inhalation exposures  have  not been




reported.








2.3.4.  Chromium  Carcinogenesis.   It  is presumed  that all  forms of Cr(VI) are




carcinogenic but the degree of carcinogenicity is  modified  by the solubility of




the  specific  compounds.   Using the International Agency for Research on Cancer




(IARC)  criteria,   animal  studies  have provided  sufficient  evidence  for the




carcinogenicity of  the following Cr(VI) compounds:   calcium chromate, strontium




chromate,  and zinc  chromate.   Both Cr(lII)  and Cr(VI)   compounds  have been




ineffective in producing lung tumors by inhalation in animals.   Similarly, nega-




tive  results  have  been obtained following the ingestion of Cr(III).   Chromium




has,  however,  been shown  to  be carcinogenic  by intrabronchial, intrapleural,




intramuscular implantation,  or subcutaneous  injection.   Cr(III) compounds have
                                      2-5

-------
been studied less extensively than Cr(Vl);  however,  animal studies indicate that




Cr(VI) is more likely to be the etiologic agent in human chromium-related cancer.




     The epidemiologic studies of chromium have demonstrated an association with




respiratory  cancer  in  chromate-producing  industries.   The  strength  of  the




association  is  evidenced  by  the  high  relative  risks of  lung  cancer  and  the




consistency  of  results  by  different  investigators  in  different  countries.




Results of three epidemiologic studies of chrome pigment workers  are also sugges-




tive of an association with lung cancer.   Less clear,  however,  is the question of




which  form of  chromium is  carcinogenic.    One  epidemiologic study of  chrome




pigment workers  (Davies,  1978,  1979)  suggested that zinc chromate  was  carcino-




genic,  while  lead  chromate was  not.   The  data on  the  lead chromate  pigment




workers, however, were  limited by smal]  sample size.   Most  of the epidemiologic




studies did  not attempt to distinguish the  carcinogenic species  of  chromium.




     Using the  IARC  criteria,  epidemiologic  studies provide sufficient evidence




that  chromium  is  a human  carcinogen;  also,  the  animal  bioassay  studies  have




provided sufficient  evidence  for  the carcinogenicity of Cr(VI).   The  carcino-




genic  evidence  of  Cr(III)  is  inconclusive.  Cr(VI)  is  mutagenic  in  multiple




tests while the data for Cr(lII) is inconclusive.




     Using the  IARC classification scheme,  the  level of carcinogenic  evidence




available for the combined animal and  human data would place chromium in Group 1,




meaning  that  there  is  decisive  evidence  for  the   human  carcinogenicity  of




chromium.









2.3.5   Dermatological   Aspects of  Chromium.   Chromium  reactions  of   the  skin




can be classified as follows:  primary .irritations,  including ulcers (corrosive)




reactions), scars,  and nonulcerative  contact dermatitis; and allergic contact




dermatitis, including  both eczematous and  noneczematous (NAS,  1974).   Contact
                                    2-6

-------
 dermatitis  can be caused by  both  Cr(III) and Cr(VI)  compounds, although most




 cases  reported are attributed to exposure to Cr(VI).








 2.3.6. Chromium as an Essential Element.  A number of clinical symptoms appear  in




 both  humans  and  animals  maintained on  chromium  deficient diets.   Symptoms




 reported  in humans consist  of glucose intolerance, weight loss, and  confusion,




 while  in  animals,  decreased fertility,  corneal opacity and  aortic plaques have




 also been observed.  The symptoms  in humans have  been reported to be reversed  in




 some cases  by  supplementing the diet with chromium  (III).  Individuals prone  to




 chromium deficiency include the elderly,  diabetics,  pregnant  women, malnourished




 children, individuals  who are offspring or siblings  of diabetics,  and persons




 with early coronary heart disease.  The Estimated  Adequate and Safe Intake  (EASI)




 level  for chromium ranges between 0.01  to 0.20  rag/day, depending on age (NAS,




 1980), while  the  average human  intake of chromium  is  17  ng/day  (range of 0  to




 224 jig/day)  (NAS,  1980).   There  is at present  no Recommended  Daily Allowance




 (RDA) for this metal.








 2.4. HUMAN BIOLOGICAL MONITORING








 2.4.1.    Chromium in Blood.  Chromium is absorbed  through both  the respiratory




 tract and gastrointestinal  system  (U.S. EPA,  1978).  Exact values for chromium




 absorption from the digestive tract are not known.  Cr(III)  is poorly absorbed,




whereas chromate is better absorbed (Mertz,  1969).




     In the respiratory tract, water and serum soluble chromium(IV)  is absorbed



into the blood system,  whereas insoluble Cr(III)  precipitates, particles and the




inert oxides and hydroxides of Cr(III) have long residence times in  lung tissue



 (U.S.   EPA,  1978).
                                      2-7

-------
     Once in the blood stream, chromium compounds are bound by proteins  (Gray and




Sterling, 1950).  It  has  been shown  that  ionic Cr(Vl) (injected intravenously)




passes through the membrane of red blood cells and  binds  to the globin moiety of



hemoglobin.   Hopkins and  Schwartz  (1964)  reported that,  in  physiological




amounts,  cationic  Cr(III) is bound  to siderophilin  and transported  to other




tissues.




     Chromium disappears quickly from the  blood and is taken  up by other tissues




in the body, where it is concentrated  much more heavily  (by a factor of 10 to 100)




than in  the blood.  Therefore,  blood levels of chromium are not a usable indi-




cator of chromium nutritional status (Mertz, 1969;  Mertz and Roginski,  1971).








2.4.2.  Chromium in Urine.  A wide range of  values  for chromium content in urine




has been reported.   Hambidge  (1971) reported chromium  levels in urine of 8.4 ppb




for adults and 5.5 ppb for children over a 24-hour  period.  Imbus et al.  (1963)




reported median urinary concentrations of chromium  for adult males of 3.77 |ig/&.




Renal  excretion  is the  major  pathway  of  chromium elimination,  with  >80%  of




injected chromium excreted in this manner (Mertz, 1969).




     Franchini et al.  (1975) and Borghetti  et al. (1977) reported  on workers




exposed to chromium  in the chromium-plating industries. They showed that urinary




excretion and renal  clearance of diffusible  chromium were two biological indices




to evaluate the degree of current exposure and the body burden of the compound,




respectively.  Franchini et  al.  (1978)  confirmed their earlier  results with an




experimental investigation using rats.




     Other authors have demonstrated  a close relationship between the amount of




Cr(VI) in the  air and  urinary excretion  (Gylseth et al., 1977) or urinary excre-




tion of the metal corrected for  creatinine (Tola et al.,  1977).   Gylseth et al.




(1977) reported  in  an  abstract  that welders  exposed to  a concentration  of
                                      2-8

-------
0.05 mg/m^  (measured  as  chromium)  had  a  urinary  chromium concentration  of


=40 \ig/i, measured after work.  The duration of exposure was not reported.




2.4.3.    Chromium in Human  Hair.   Schroeder and  Nason  (1969)  reported a mean


chromium concentration of 0.69 + 0.063 ppm for women.   Hambidge et al. (1972)


measured chromium concentrations at various distances from the hair root.  They


reported that variation in the concentrations were due  to past fluctuations in


chromium  nutritional  status.   Hambidge and  Rodgerson  (1969)  reported higher


levels of chromium in the hair of nulliparous women (0.2 to 2.81  ppm) than in the


hair of parous women (0.04 to 1.14  ppm).  However, a later study by Hambidge and


Droegnueller (1974)  found  changes in hair chromium levels  due  to  pregnancy not to


be statistically significant.  Hambidge and Rodgerson (1969) reported that hair


chromium levels in 3- to  8-month-old  infants  were significantly higher than in


those of 2- to 3-year-old children.   By  the second year of life, mean chromium


levels in hair approached values present in older humans.





2.5. HUMAN HEALTH RISK ASSESSMENT OF CHROMIUM





2.5.1. Health Effects Summary.  Although a number of epidemiologic studies have


found an association between exposure  to  chromium and lung cancer, the data that


could be used  for estimating the cancer risk due to exposure  to  chromium are


limited to the study of Mancuso (1975). Mancuso  (1975) reports age-specific lung


cancer mortality  data for  chromate production workers in terms of total elemental


chromium exposure.  Using this  information, CAG  estimated  the  lifetime cancer

                                                        q
risk due to a constant exposure  to  air containing 1 jig/nr of elemental chromium
                                      2-9

-------
to be 1.2 x 10 2.  This is considered an upper-bound estimate, since it




is based on a model that is linear at low doses.







2.5.2  Populations at Risk.  There is no toxicological information to




indicate that any specific subpopulation is highly sensitive to the toxic




effects of chromium.  An additional burden of chromium may result from




environmental sources in individuals exposed to chromium in the workplace,




although predominant exposure would be from the workplace in most instances.




The National Institute for Occupational Safety and Health (NIOSH, 1975)




has estimated that 175,000 workers are exposed to Cr(VI), and 200,398




workers are exposed to Cr(III).  The same individuals may be represented




in both groups of employees.
                                   2-10

-------
                           3.  BACKGROUND INFORMATION








3.1.  CHEMICAL AND PHYSICAL PROPERTIES




     Chromium was  discovered  in 1797  by  the French  chemist,  Louis Vanquelin.




Metallic chromium is steel gray  in color, melts at 1857 + 20°C, boils at 2672°C,




and has a specific  gravity of 7.20  at 28°C (Weast,  1980).  As found in nature,




chromium is  a mixture of four stable  isotopes of mass numbers 50, 52,  53, and 5*1.




     The inorganic chemistry of chromium and its compounds has been extensively




studied.  However, its  physical  or chemical  forms and  the mode by which they are




incorporated  into  biological   systems are  poorly  characterized.    Inorganic




chromium compounds occur  in valence  states  ranging  from -2 to  +6.   In natural




systems, the Cr(III) and Cr(VI) states are the two most stable forms of chromium.




     Chemically,  the Cr(III)  state  is  the most stable  and  important  form of




chromium.   In neutral  and  basic  solutions, Cr(III)  forms  binuclear and poly-




nuclear compounds in which adjacent  chromium atoms  are linked through hydroxy-




(OH) or  oxo-(0) bridges.   Interestingly,  Cr(III)  forms  stable complexes with




amino acids and peptides  (deMeester and Hodgson, 1977; deMeester et al.,  1977).




Cr(III) also has a strong tendency to  form hexacoordinated octahedral complexes




with ligands, such as water, ammonia, urea, ethylenediamine, halides, sulfates,




and  organic acids.   These relatively stable  complex formations  (Cotton and




Wilkinson,  1962) can prevent precipitation  of Cr(III) at  pH values at which it




would otherwise precipitate.




     Cr(VI)  exists in solution as hydrochromate, chromate, and dichromate ionic




species.  The proportion of each ion in solution  is dependent on pH.  In strongly




basic and neutral pHs,  the chromate form predominates. As the pH is  lowered, the




hydrochromate concentration increases.  At very low pHs, the dichromate species
                                      3-1

-------
predominates.  In  the  pH ranges encountered in  natural  water,  the predominant




forms are hydrochromate ions (63.6$)  at  pH 6.0 to 6.2 and chromate ion  (95.7$) at




pH  7.8  to 8.5 (Trama  and  Benoit,  1960).  The  oxidizing ability  of  Cr(VI) in



aqueous solution is pH  dependent.  The oxidation potential of Cr(VI) increases at




lower pHs.  The ability of Cr(VI) to oxidize organic materials and the tendency




of  the  resulting Cr(III)  to form stable  complexes with  available  biological




ligands afford a reasonable  mechanism  by which chromium can interact with the




normal biochemistry of man (Towill et al., 1978).




     The physical properties of a few environmentally significant chromium com-




pounds are shown in Table 3-1.  The same parameters  for the Cr(VI) compounds are




shown in Table 3-2.  It should be mentioned that there is considerable disagree-




ment in the literature  concerning the physical parameters  given in Tables 3-1 and




3-2  and  therefore  these values  should  be  accepted  with  reservation.    The




disagreement in the values is possibly  due to  the  reactions of these compounds




with other substances,  namely with moisture and air at high temperatures, impuri-




ties, and structural and  compositional changes occurring during the experimental




determinations.




     Reactions of Cr(III) in alkaline solution depend on the concentration of OH"




ions.  When hydroxyl ions are added  to  Cr(III)  solutions  in quantities insuffi-



cient for precipitation,  basic ions are  formed, which  then  polymerize with the




formation of hydroxy- or  oxy-bridged  compounds  as shown below:




                                  3+                       2+
oi u»u_;_ 	 f 01 ^ripu;/- j
(in solution)
F ( H n } Pr^n^rrfii ni i •<- 	 r
L viipU yp-Vji — u— oi ^iipUV(-J L
(oxolation)
	 ^ L^r^uHMHpu;,-J
°H" i
A standing
2 OH 2 4
(olation)
                                     3-2

-------
U)
                                                                           TABLE 3-1

                                                  Physical Properties of Selected Trivalent Chromium Compounds1
Compound Formula
Chromic acetate Cr(CH.COO) • H 0
332
Chromic chloride CrCl,

Chromic chloride, [CKH.OkCl-lCl • 2H-0
hexahydrate <2 1 Z 2
[Cr(H20)6]Cl3
Chromic formate, [Cr(HCOO) ] • 6H-,0
hexahydrate * i
Chromic oxide Cr_0,
Chromic phosphate, CrPO,. • 2H-0
hydrated CrPO,j • 6H20
Chromic sulfate Crp(SOj.)-
Chromic sulfate, Cr,(sok}_ • 15H 0
hydrated 2 M 3 2
Cr2(SO^)3 • 18H20
Density,
g/cm-5
NR
2.76 (15»C)

1.76
NR
NR
5.21
2.U2 (32.5°C)
2.121 (1lt°C)
3.012
1.867 (17°C)
1.7 (22»C)
Melting
Point, °C
NR
«1150

83
NR
decomposes
above 300
2266
NR
100
NR
100
-12H20, 100
Boiling
Point, °C
NR
1300
(sublimes)
NR
NR
NR
4000
NR
NR
NR
-10H20, 100
NR
Solubility
in Water, g/ 100 mi
slightly soluble
insoluble

58.5 at 25»C
soluble
soluble
insoluble
slightly soluble
insoluble
insoluble
soluble
120 at 20 »C
   •Source:  Weast,  1980; The Merck Index, 1976
   NR = Not reported

-------
                                                                       TABLE 3-2

                                             Physical Properties of Selected Hexavalent Chromium Compounds8
Compound Formula
Ammonium chromate (NHj^)2 CrOj.
Ammonium di chromate (NHj.)2Cr207
Barium chromate BaCrOj.
Chromium (VI) oxide CrO_
Lead chromate PbCrOj,
Hercurous (I) chromate HgjCrOjj
M«r curie (II) ohromate HgCrOj.
Potassium chromate K-CrOj.
Potassium dichromate K2Cr20_
Sodium ohromate Ma CrOj.
Sodium dichromate Na^CrpO- • 2H-0
dihydrate
Density, b
g/'cm3
1.9112
2.15525
4.49825
2.7025
6.1215
NH
NR
2.73218
2.67625
2.72325
2.34825
Melting
Point, °C
180
decomposes
180
decomposes
decomposes
197
844
decomposes
decomposes
971
398
792
84.6
(incongruent)
Boiling
Point, "C
NR
NR
NR
decomposes
decomposes
NR
NR
NR
500
decomposes
NR
400
decomposes
Solubility in
Water, g/100 mi
40.5 at 30»C
30.8 at 15»C
3.4 x ID'1*
at 160»C
67.45 at 100°C
5.8 x 10~6 at
25 *C
very slightly
soluble
slightly soluble,
decomposes
62.9 at 20»C
4.9 at 0°C
102 at 100»C
87.3 at 30»C
180 at 20 "C
 Source:  Weast, 1980; Hartford, 1979
bThe lower figures indicate the tenperature («C) at which the densities were measured.
NR = Not reported

-------
Such ions are  of  the proper size to cross-link protein  fibers  and may play an


important part in the chemistry of tanning.   The  single  hydroxyl-bridged rhodo


and erythro binuclear Cr(III) amine complexes  also have been extensively studied


(Veal et al., 1973;  Cline et al.,  1981).


     When a  sufficient  amount  of a  base  is added to Cr(III) salt solution,  a


hydrous oxide of indefinite compositipn, Cr2Og'xH20, is precipitated.  On addi-


tion of more base  to the hydrous oxide,  the precipitate redissolves, probably due

                                                       O_Y                  _
to  formation  of  complex  ions of  the  type  [Cr(OH)x3     (e.g.,  [Cr(OH)^] ,


[Cr(OH)6]3").


     Cr(III)  compounds   are reduced  to  Cr(II)   compounds by  hypophosphites,


electrolysis, or reducing metals,  such as Zn, Mg, and Al  in acid solution.  In

                                                     2_
basic solution, Cr(III)  is readily  oxidized  to  CrCK    by hypochlorite, hypo-


bromite, peroxide, and oxygen  under  pressure at  high temperature.   Heating of


chromium compounds in air in the presence of alkalies also yields chromate.  In


acid solution, Cr(III) is harder to  oxidize  and needs  strong  oxidizing agents,


such as concentrated HClOj., sodium bismuthate, and permanganate.


     All Cr(VI) compounds except CrFfi are oxo-compounds.  Cr(VI) rarely occurs in


nature, apart from anthropogenic sources,  because it is  readily reduced in the


presence of  oxidizable organic matter.    However, after  it is  introduced into


water, Cr(VI) frequently  remains unchanged in  many natural  water sources because


of low concentration of reducing matter.  Cr(VI) occurs most commonly in the form


of  chromate  or dichromate,  both of  which are  produced  on  a  large  scale in


industry.  The dissociation equilibrium of  chromic  acid  solution indicates the


weaker acidity of E^CrO^.   The dissociation  of HpCr-O., appears  to be that of a


strong acid.  Acid solutions of dichromate are powerful oxidizing agents:
                                      3-5

-------
     2_
Cr>2°7   + 14H+ + 6e~ - > 2Cr+3
                                             H20, E° =  1.33V

 Dichromate  salts  are  the leading commercial form of Cr(VI).


 3.2.  PRODUCTION, USE, AND RELEASES TO THE ENVIRONMENT


 3.2.1.   Production of Chromium  Compounds.  The  industrial  processes  for the

 production  of chromium metal and the various  compounds have been described by

 Hartford  (1979).  A  simplified  flow chart depicting these processes appears in

 Figure  3-1.

     There  has  been  no mining of  chromite ore in the United States since  1961.

 Chromium  ore  and  ferrochrome  alloys  are  imported mainly from the Soviet Union,

 South Africa, Turkey, and Zimbabwe.

     The  two  primary industrial compounds of chromium made directly from chrome

 ores are  sodium chromate  and sodium dichromate.  Secondary chromium compounds

 produced  in  substantial quantities  include potassium  chromate  and  potassium

 dichromate, ammonium dichromate, chromic  acid,  and various formulations of basic

 chromic sulfate used principally for leather tanning. The United Stated manufac-

 turers of the three  important  chromium compounds, their  production capacity, and

 the amount produced are discussed below.

     The manufacturers of both sodium chromate and  sodium  dichromate  and their

 annual production capacity in  1982 are given in  Table 3-3.  The estimated produc-

 tion capacity is based on a 100? sodium dichromate weight.

     The manufacturers of chromic acid and their annual production capacity in

 1982 are given in Table 3-4.


3.2.2.    Uses of  Chromium  and Its  Compounds.   The  United States  consumption

pattern of chromium  and its  compounds for the  year 1979  is  shown in Table 3-5.
                                      3-6

-------
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-------
                             TABLE 3-3

         Manufacturers and Their Production  Capacities  of
              Sodium Chromate and Sodium Bichromate3
                                     Annual  Production Capacity
        Manufacturers                in 1982,  10^ metric tons
Allied Chemical Corp.                         65  (59)
 Baltimore, MD

American Chrome and Chems.,  Inc.              45  (41)
 Corpus Christi, TX

Diamond Shamrock                             94  (85)b
 Castle Hayne, NC

    TOTAL                                  204  (185)


  Source:   SRI  International,  1982;  1982 U.S. Industrial Outlook,
  Chemical Marketing Reporter,  1982.

   Diamond  Shamrock  will   increase  capacity  to  118,000  tons
  (106,200 Mg) in January  1983.
                            3-8

-------
                       TABLE  3-4
                                                     a b
Principal United States Manufacturers of Chromic Acid '
                           Annual  Capacity  in  1982,
                                 •3            c
    Manufacturers              10  metric  tons
 Allied  Chemical  Corp.                21
  Baltimore, MD

 Diamond Shamrock Corp.               24
  Castle Haynes,  NC
  Source:   SRI  International,  1982;  Hartford,  1979

  Data on actual production of chromic acid are held to be
  confidential    to    avoid    disclosing    proprietary
  information on individual  companies.

 °The  estimates for  production  capacity  are  based on  a
  100? chromic  anhydride  (CrO_)  basis.
                          3-9

-------
                                    TABLE 3-5

               United States Chromium Consumption Pattern in 1979a
                                   Quantity Consumed13'0          % Fraction of

                                     10^ metric tons           U.S. Consumption
Metallurgical
  wrought stainless and heat             235                          44
     resisting steels
  tool steels                              6                           1.1
  wrought alloy steels                    44                           8.1
  cast alloy steels                       15                           2.7
  alloy cast irons                         8                           1.4
  nonferrous alloys                       15                           2.7
  other                                    6                           1.1
    Total                                329                          61.0

Refractories
  chrome and chrome-magnesite             16                           3.0
  magnesite-chrome brick                  23                           4.2
  granular chrome-bearing                 42                           7.8
  granular chromite                       16                           3.0
    Total                                 97                          18.0
Chemicals
pigments
metal finishing
leather tanning
drilling muds
wood treatment
water treatment
chemical manufacture
textiles
catalysts
other
Total
Grand Total

29
24
18
5
7
7
9
4
<2
9
114
540

5.4
4.4
3.3
0.9
1.3
1.3
1.7
0.7
0.3
1.7
21.0
100
aSource:   Hartford, 1979,  Mineral Commodity Summaries, 1980.

 Exclusive of scrap.

 Columns  may not total exactly due to rounding.
                                    3-10

-------
     It  can  be  seen from  Table 3-5  that  metallurgical  and  chemical  usages




constituted  82$ of  the  total United States  chromium  consumption in  1979.




Metallurgical grade chromite ore is usually converted into one of several types



of ferrochromium or  other chromium metal that  are  alloyed with  iron  or other




elements,  such  as  nickel  and cobalt.   A great  variety of  useful  steels are




produced from these alloys.   Because  of  their high  melting points and chemical




inertness, chromite ore and chrome alloys are used by the  refractory  industry in




furnaces  as  linings, in  the  manufacture of  furnace   bricks,  and  as  coating




materials  to close pores  and  to join bricks within the furnace.  Other uses of




chromite  refractories include  nonferrous  alloy  refining,  glass making,  and




cement processing.  The pattern of chromium consumption in the United States has




been consistent over the last  20 years.  However,  the use  of chromite and chrome




alloys in the refractory industry is  beginning to decline as open hearth furnaces



are replaced by basic-oxygen furnaces  (Hartford, 1979).  In the future, growth in




chromium usage  is expected  in the metallurgical and chemical  sectors.  A recent




study  (Morning, 1977) has projected a  3.4$ growth annually in the United States




chromium  consumption, leading  to a  total chromium  demand  of 1  million tons




(900,000 metric tons) in  the  year 2000.



     The consumption pattern of imported chromium  for metallurgical,  refractory,




and chemical usage is shown in Table 3-5.  Chromium-containing pigments can be




primarily  classified into chromate   color  pigments   based   on lead chromate,




Cr(III) oxide greens, and corrosion inhibiting  pigments based primarily on zinc




chromate.  In metal finishing, chromic acid is used  in  chromium  plating of metal




surfaces.  The  chrome tanning of leather  is one step in a complicated series of




operations leading from the rawhide to the finished product.  The annual consump-




tion of hides by the leather industry is decreasing (Hartford,  1979),  and  the use




of  Cr(VI) compounds  for tanning  purposes may be  on  the decline.   Chromium
                                      3-11

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chemicals, such as chromium lignosulfonates, are used in drilling muds during the




drilling  of wells  to  combat  fatigue  corrosion  cracking  of  drill  strings



(Hartford, 1979).  Chromated copper arsenate is widely used as a wood preserva-



tive, especially in treating  utility poles, building lumber,  and  wood founda-




tions.  Chromates are  used to inhibit  metal corrosion in recirculating water



systems,  such  as  cooling  towers,  locomotives,  and  automobiles.    Sodium



dichromate and  various  chromic  salts are  employed  in the textile  industry to



improve washfastness and to oxidize the  dyed textile.  A large number of chromium



compounds  are  used  as  catalysts  in various  chemical reactions.   Barium  and



calcium  chromates  are   used  as  activators  and dipolarizers  in  fused  salt



batteries.  Chromium dioxide (CrO?) is used as a ferromagnetic material in high-



fidelity magnetic tapes.







3.2.3.   Releases to the Environment.    Although  the chromium industry  in the



United States  has adopted various pollution control  measures, some release of




chromium compounds into  the environment is  occurring.  Chromium compounds from



industrial operations enter the environmental air, water, and soil from several



sources.    Kilns,  smelting  furnaces,   boilers,  leaching  tanks, open boiling



vessels, plating tanks, and other installations emit dusts and mists containing



chromium to the atmosphere.



     Chromium  is a trace component of coal  and oil and  is released  to the



atmosphere upon combustion of these fuels.  Fly ash emitted from coal-fired power




plants contain  10 to  600 ppm chromium,  depending on the type of boiler firing



(Block and Dams, 1976).   For power plants equipped with electrostatic precipita-



tors (ESP) to prevent particulate  emissions,  the total chromium concentration in




the emission can be reduced (NAS,  1971*).  Rinaldi et al. (1980) have shown that



the chromium concentration  of particulates  from controlled coal combustion may
                                      3-12

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be higher than the  chromium concentration of particulates from uncontrolled coal
combustion, due  to preferential  chromium enrichment  on small  particles  that
escape the ESP device.
     Wood contains chromium and it  is  likely  that the burning of wood in fire-
places and campfires may  contribute small  amounts of  chromium  into the atmos-
phere.  Forest fires would therefore be a potential non-anthropogenic source of
atmospheric chromium (NAS, 1971*).  No estimate of the amount of chromium emitted
from  forest  fires  could  be  obtained.   However,  Lantzy and MacKenzie (1979)
estimated  chromium flux in the atmosphere  from anthropogenic  (industrial and
fossil  fuel)   and  non-anthropogenic  (continental  dust,   volcanic dust,  and
volcanic gas) sources.  The ratio of anthropogenic to non-anthropogenic atmos-
pheric flux of chromium  was  estimated to be 1.61.   Incineration of municipal
refuse and sewage sludge is also expected to contribute  small amounts of chromium
into the atmosphere (Rinaldi et al., 1980; Fiscus et al., 1978).
     Asbestos mining is another source of  chromium emissions  to the atmosphere
since  asbestos  has been  found to  contain  as  much  as  0.15?  chromium (Towill
et al., 1978).  The wearing of vehicular brake linings, therefore, represents a
potential  source of chromium  in  the  atmosphere.   Catalytic  emission control
systems  in automobiles  using  copper  chromite  reduction  catalysts  represent
another source of chromium emissions to the atmosphere (IARC,  1980).  Also, when
chromate  chemicals are used as corrosion  inhibitors  in  recirculating cooling
waters, some chromate is  lost to the atmosphere as mist.
     According to a GCA Corporation (1973) report, United States industrial and
inadvertent sources of  chromium emissions  into  the atmosphere under controlled
operations amounted to 18,200 metric  tons in  the year  1970.   The  amount of
atmospheric chromium  emissions from  different  sources is  shown in Table 3-6.
Although  the  total estimated chromium  emissions  given  by  the  two  studies in
                                      3-13

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                                   TABLE 3-6
                     Sources and Estimates of United States
                    Atmospheric Chromium Emissions  in  1970a

Chromium
Emissions, metric
GCA Estimates
Source
Industrial Sources:
refining
steel and alloy
material handling
chemical processing
refractory
Inadvertent Sources:
coal combustion
oil combustion
cement production
incineration
Asbestos Mining
Total
Uncontrolled

18,700
2,407
1,100
835
4,784

7,900
336
NR
NR
9
36,100
Controlled

11,200
595
750
106
1,650

1,420
336
254
143
0
16,500
tons/ year
Goldberg
Controlled

3,800
NR
NR
NR
6

7,030
69
NR
NR
7
10,900
 Source:   GCA Corporation,  1973;  Goldberg,  1973
NR = Not reported
                                   3-14

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Table 3-6  are  somewhat  comparable,  there  is  a  substantial  difference  in




estimated emissions from individual sources.



     The  geographical  distribution of  atmospheric chromium  emissions  in  the



United States is presented in Table 3-7.  It is evident from Table 3-7 that the



Great Lakes area,  the Southeast and the East coast south from New York constitute




the bulk of atmospheric chromium emissions in the United States.



     Chromium compounds occur in a variety of industrial wastewaters and poten-



tially  may enter  surface water  and  groundwater  supplies.    Wastewaters  from



electroplating operations, leather tanning, and textile manufacturing represent



the types  of  chromium-containing  streams  that  may ultimately enter surface and



groundwaters  (Hartford, 1979).  It has been estimated that 220 metric tons/year



are discharged in Southern California coastal waters (Schafer, 1977).  Ottinger



et al.  (1973) estimated that  6200  metric  tons  of  chromium are lost annually in



the sludge of solvent-based paints and another  437  metric tons  are discharged as




paint residues.



     Solid waste  streams  containing Cr(VI) constitute the primary problem area




involving  chromium solid wastes  (Hartford,  1979).  Wastes  resulting from the



roasting and  leaching steps in the chromate manufacturing process traditionally



contain residual  Cr(VI).   If landfilled,  the  residual  Cr(VI)  can slowly leach



into surrounding  waters via desorption and disproportionation  (Hartford, 1979).



An estimate of the total amount of chromium released into soil and groundwater as



a result  of the leaching  of chromium-containing solid wastes is not available.








3.3.  ENVIRONMENTAL FATE  AND  TRANSPORT








3.3.1.  Air.  Little information exists  in the literature regarding the nature of



the  chemical  species present  in  the atmosphere  away from  obvious  sources of
                                      3-15

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                                    TABLE 3-7

              Regional  Distribution  of Principal  Chromium Emissionsa
EPA Region/States
I/CT,MD,MA,NH,RI,VT
II/NJ,NY,PR,VI
III/DE,MDfPA,VA,WV,DC
IV/AL,FL,GA,KY,MS,NC,SC,TN
V/IL,IN,MI,MN,OH,WI
VI/AR,LA,NM,OK,TX
VII/IA,KS,MO,NB
VIII/CO ,MT , ND ,SD ,UT , WY
IX/AZ,CA,NV,HA
X/AK,ID,OR,WA
TOTAL
Annual Chromium Emissions
From Sources in this Region,
metric tons (Mg)b
103
3,1^0
2,800
3,000
4, 830
74
177
107
174
898
15,300
Percent of Total
U.S. Chromium
Emissions
0.6
19.0
17.3
18.5
29.0
0.5
1.1
0.7
1.1
5.4
93.2
 Source:   GCA Corporation,  1973

 Sources  include ferrochrome production,  refractory production,  cement production,
chrome steel production,  and coal  and  oil combustion.
                                        3-16

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pollution.  Under  normal  conditions,  Cr(III) and  Cr(0)  in the  air  should not



undergo any reaction  since these species are chemically inert (Towill et al.f



1978).   Cr(Vl)  in air could  eventually react with  dust particles  or  other



pollutants  to  form Cr(III)  (NAS, 197*0.   However,  the exact  nature  of such



atmospheric reactions has not been extensively studied.



     Low concentrations of chromium enter the  atmosphere as a result of  indus-



trial activities and  soil-derived aerosols  (Towill et al., 1978).  Chromium is



removed from  air by  atmospheric  fallout and precipitation.   The total  yearly


                                                            2          2
deposition of chromium in  urban areas may vary from  0.12 ug/m  to  3 Hg/m  (Towill



et  al.,  1978).   Depending upon  the  locale, between 44 and  96? of the total



deposition occurs by wet precipitation.  In  general,  urban  areas  have higher wet



and total deposition  than rural areas.   Chromium concentration in a wet deposi-



tion may vary from 0.004  to 0.060 \ig/mi  and 0.0006  to 0.034  (ig/fi, for urban and



rural areas, respectively (Towill et  al., 1978). The precipitated chromium from



the air enters the surface water  or soil.



     Chromium  particles of aerodynamic  equivalent  diameter  <20 [im may  remain



airborne for long periods  and may be transported great distances by wind currents



and  diffusion  forces  (Sehmel   and  Hodgson,   1976).    Therefore,  atmospheric



conditions  play  an important  role in  determining  the  chromium concentration



around emission sites; however,  no data relating atmospheric chromium content to



atmospheric or meteorological conditions could  be  found  in the literature.







3.3.2.  Water and Sediments.  Surface  runoff, deposition  from air, and release of



municipal  and  industrial  wastewaters  are  the  sources  of  chromium  in surface



waters.   Chromium may be transported  in five  forms  (see Table  3-8) in surface



waters.
                                      3-17

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                                   TABLE  3-8

       Five Forms of Chromium Transported in the Yukon and Amazon Rivers*

      ~~Percent  Present Tn
          Physical Form                   Amazon River    Yukon River


In solution and organic complexes             10.4              12.6

  Adsorbed                                    3.5              2.3

Precipitated and co-precipitated              2.9              7.2

  In organic solids                            7.6              13.2
  In sediments                               75.6              64.5


•Source:   Towill et  al.,  1978
                                     3-18

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     It can be seen from Table 3-8  that  most  of the chromium in surface waters




may be present in particulate form as sediment.  Some of the particulate chromium



would  remain  as suspended  matter  and  ultimately  be  deposited  in  sediments.



DeGroot and Allersma (1973) found a chromium ratio in water to suspended matter



to be 1 to 2.3 for the Rhine River.   In the heavily polluted Qishon-Gadura River



in  Israel,  chromium  concentrations  in  water  were  <10  ppb,  while  sediment



contained from 220 to 610 ppm chromium (Towill et al., 1978).



     The exact chemical  forms of chromium in surface waters  are not well defined.




Although most of the soluble chromium in surface water may be present as Cr(VI)




(Towill et al.,  1978), a small amount may be present as Cr(III) organic complexes



(DeGroot and Allersma, 1978; Fukai,  1967).  Schroeder  and Lee  (1975) studied the



transformation between  Cr(III) and  Cr(VI) in natural waters.   They  found that



only 3% of Cr(III) was oxidized by 0- in 30 days at ambient temperature.  Cr(VI)



is the major stable form of chromium in seawater (Fukai, 1967); however, Cr(VI)



may be reduced to Cr(III) by organic matters present in water and may eventually




deposit in sediments.  Lu and Chen  (1976)  found that  chromium was not signifi-



cantly released from sediments into seawater under either  oxidizing or reducing




conditions.








3.3.3.  Soil.  Most soil chromium is in mineral,  absorbed,  or precipitated form.



Chromium probably occurs as the insoluble Cr(III) oxide  (Cr^O-j'nHpO) in soil, as



the  organic matter  in  soil  is  expected  to  reduce any  soluble chroraate to



insoluble Cr?0o. Chromium in soil can be transported to  the atmosphere by way of



aerosol formation  (John et al.,  1973; Zoller  et al.,  197*0«   Chromium is also



transported from soil through runoff and  leaching of water.  Runoff could remove




both  chromium  ions  and  bulk precipitates  of  chromium with final deposition on



either a different land  area or a water body.  In addition,  flooding of soils and
                                      3-19

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 the subsequent anaerobic decomposition of plant matters may increase  dissolution


 of Cr(III)  oxides in  the soil  (Towill et  al.,  1978).





 3.4.  LEVELS OF CHROMIUM IN VARIOUS MEDIA





 3.4.1.  Ambient Air.  The chemical form of chromium in air depends  on the source


 of emission.  The vast majority of chromium in the atmosphere, originating from


 such  sources as metallurgical  production, coal and oil combustion,  cement pro-


 duction, and incineration,  is usually in the Cr(III) or Cr(0) state.  However, a


 small amount  of  total chromium in the  atmosphere may be present in the Cr(VI)


 state.  Chrome production, chrome plating, and cooling  tower drifts  are primary


 examples of the sources of Cr(VI) in the atmosphere (Towill et al.,  1978).  The


 mass median diameter of chromium in air particulate matter is in the range of 1.5


 to 1.9  ^im  (Cawse,  1974;  Lee and von Lehmden,  1973).   Chromate salts are often


 used  in the cooling tower  water  as a corrosion  inhibitor.   Therefore, cooling


 tower drift consisting of  water  droplets formed mechanically within the towers


 and carried by wind into the  surrounding  area may be a  source  of  high Cr(VI)


 concentrations in air.  Air  concentrations of chromium near a cooling tower at


 the Oak Ridge Gaseous Diffusion Plant, Oak Ridge, Tennessee, were about 50 ng/m^


 from distances up to 660 feet  (200 m) from the tower  (Alkezweeny et  al., 1975).

                                              p
 Hourly  chromium  deposition  was  about  1 mg/m   at 100 feet  (30 m)  and  about


 0.01  mg/m  at 330 feet (1000 m) from the tower.  Therefore, substantial amounts


 of chromium may be  present in the respirable particulate fraction  and  can be


 deposited in the respiratory tract.


     The concentrations of  total  chromium measured in  the  ambient  air of many


 urban and nonurban areas of the United States  during  1977 to  1980  are given in


Table 3-9.   The  data in Table 3-9 were obtained from  the  U.S.  EPA's National
                                     3-20

-------
TABLE 3-9
Total Chromium Concentrations Measured in the Ambient Air
of Selected Sites in the United States During 1977-1980
Site
Grand Canyon, National Park, AZ
Los Angeles, CA
Water bury, CT
Atlanta, GA
Hawaii County, HId
Kansas City, KS
Iberville Parish, LA
Acadia National Park, ME°
Baltimore, MD
Worcester, MA
Bayonne, NJ
Newark, NJ
Niagara Falls, NY
Year
1977
1977
1978
1979
1977
1980
1977
1977
1978
1980
1977
1978
1980
1977
1977
1979
1977
1978
1979
1977
1978
1980
1978
1979
1980
1979
1980
Total Chromium Concentration, ^g/m
Arithmetic Maximum Observed
Mean5 Value0
0.0058
0.0188
0.0342
0.0326
0.0089
0.0062
0.0063
0.0167
0.0276
0.0191
0.0063
0.0059
0.0052
0.0052
0.1568
0.0935
0.0063
0.0099
0.0067
0.0105
0.0149
0.0123
0.0181
0.0129
0.0091
0.0389
0.01411
0.0134
0.0666
0.2178
0.1396
0.0441
0.0194
0.0216
0.0413
0.0724
0.0358
0.0159
0.0128
0.0052
0.0052
2.4870
0.4589
0.0167
0.0239
0.0166
0.0253
0.0324
0.0508
0.0301
0.0333
0.0369
0.5590
0.0603
  3-21

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                                 TABLE  3-9  (cont.)
Site
                                              Total  Chromium Concentration,
                                              Arithmetic      Maximum Observed
Year
 Mean
                                                    b
Black Hills National Forest,  SD
1978
0.0090
  Value"
Akron, OH



Cincinnati, OH



Steubenville, OH

1977
1978
1979
1980
1977
1978
1979
1980
1978
1979
0.0126
0.0188
0.0166
0.0204
0.0083
0.0116
0.0451
0.0150
0.0517
0.1212
0.0610
0.0528
0.0389
0.0710
0.0377
0.0294
0.4316
0.0718
0.2602
0.6839
0.0295
Chattanooga, TN



Norfolk, VA



Tacoma, WA


1977
1978
1979
1980
1977
1978
1979
1980
1977
1978
1980
0.0122
0.0140
0.0112
0.0150
0.0067
0.0069
0.0083
0.0119
0.0099
0.0249
0.0104
0.0453
0.0463
0.0760
0.0705
0.0152
0.0158
0.0291
0.1456
0.0330
0.1425
0.0283
 Source:  Unpublished data in the  National Aerometric Data  Bank maintained  by  the
Monitoring and Data Analysis Division of EPA,  Research Triangle Park, NC.
 Values represent annual average.

°Values represent maximum 24 hour  averages.
d
 Background sites; all other sites are determined  to be  populated urban  areas.
                                              3-22

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Aerometric Data Bank, which  is  maintained by the Agency's  Monitoring and Data



Analysis Division (MDAD) at Research Triangle Park,  North Carolina.  None of the



data in Table 3-9  have been previously published. The  ambient chromium data have



been collected  by monitoring networks operated  by  various  State and local air




pollution  control agencies as  required  by the  Clean Air Act.   After passing



editorial and validation checks which are performed  by EPA regional  offices, the



data are forwarded  to MDAD  for  incorporation into the National Aerometric Data




Bank.



     During  the 1977 to  1980  period, the mean  chromium concentration for the




urban areas given in Table 3-9 ranged from 0.0052 \ig/nr (the  background level) to



0.1568  (ig/m^.   The  highest mean value of 2.U87 |ig/m^  was  recorded in Baltimore,



Maryland in  1977.  For the  sites  with  >2 years worth of data,  no discernible



upward  or downward concentration trends  are evident.  For example, in Newark, New



Jersey,  the  mean chromium  concentration  dropped from 0.0181  ng/nr in 1978 to



0.0129  \ig/a^ in 1979 and to 0.0091  \ig/n? in 1980.  However, in Norfolk, Virginia,
over  the same  period  the chromium  levels rose  from 0.0069 ng/nr in  1978  to



0.0083 tig/m^  in 1979 and  to 0.0119 rng/m3 in  1980.   In Akron,  Ohio,  the  mean
 chromium concentration in  1978 was determined  to  be 0.0188  |ig/m  .  in  1979,  the




 concentration  in Akron  dropped  to 0.0116 ng/nr  but  in  1980  it rose again  to



 0.0204 ng/m^.   The mean chromium concentrations  in  nonurban,  background  areas
such as national parks ranged from 0.0052 ng/m3 to 0.0090 jig/m3 over the 1977  to




1980 period.




     Specific  industrial sources such as  power plants, incinerators,  and  iron




and steel plants may signifi'cantly increase  the atmospheric  chromium concentra-




tions found in certain areas due to the relatively high chromium content of their




emitted particulate  matter.  Lee and von  Lehmden  (1973) reported the  chromium




content of particulate emissions from coal-fired power plants to be between 1 and
                                      3-23

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100 ppm, cement plants to be between 100 and 1000 ppm,  iron and steel plants to



be  between  10 and  100 ppm,  and municipal incinerators  to be between  100  and




1000 ppm.







3.4.2.  Aquatic Media. The chromium levels detected in a few surface waters and




groundwaters are presented in Table 3-10.  The amounts of chromium found in these



waters are usually related by the authors to anthropogenic input.  For example,



it has been shown by Kopp and Kroner (1967)  that water from Lake Michigan near



industrial discharge points contained a  higher level of chromium than lake water



contaminated with lesser  industrial  input  (5  to 19 ppb, compared to 2 to 4 ppb).



     The valence of chromium in surface  water can be either VI or III.  Although



Cr(VI) is the more stable species in  seawater, Fukai (196?) provided data to show



that both Cr(III) and Cr(VI) were present.   Surface  seawater  contained Cr(III)



and Cr(VI) in the range of 0.02 to 0.14  ppb and 0.28  to 0.36 ppb, respectively,



while seawater  at  depths  of 5, 500, and  1000 m  contained  about  the same level




(0.2 ppb) of Cr(III) and  Cr(VI).



     In a survey of 14 groundwater  and 69 surface water  supplies in 83 United



States  cities  in Illinois,  Indiana, Michigan,  Minnesota, Ohio,  and Wisconsin,



the chromium level in the raw waters used for drinking waters were found to range



between <5.0 and 17.0 ppb (see Table 3-11) (U.S.  EPA,  1975).



     The chromium concentration in various United States drinking water supplies



is presented  in  Table 3-11-  In a survey of 2595 water samples  from 969 water




supplies in the United States,  only four samples showed chromium levels above the



detection limit  of  50 ppb  (McCabe et al.,  1970).  The maximum chromium concen-



tration in water detected in this survey was 80 ppb.




     In a more recent survey (1974 to 1975) with an analytical method of better



sensitivity,  3834  tap waters from 35 geographical locations  representative of
                                     3-24

-------
                                    TABLE  3-10
             Chromium Levels in a Few Surface Waters and Groundwaters*
Source
Lake Tahoe
Colorado River
Columbia River
Mississippi River
Missouri River
Ohio River
U.S. surface waters
U.S. surface waters
Natural water, Oak Ridge, TN
Water near cooling tower,
Oak Ridge, TN
Uncontaminated stream, NY
Contaminated stream, NY
Uncontaminated well , NY
Contaminated well , NY
Illinois River
Spring water, CA
Well water, CA
Stream water, CA
Seawater , CA
Frequency of
Detection %
NR
12
87
23
10
20
25
25
NR
NR
NR
NR
NR
NR
NR
6
5
0
0
Cone, (ppb or \ig/i) in Samples
with Detectable Chromium Level
Average
<0.62
NR
NR
NR
NR
NR
<1
9.7
NR
NR
<10
1250
<10
6000
21
NR
NR
NR
NR
Range
<0.07 to <0.91
10 to 30
1 to 10
3 to 20
8 to 10
4 to 16
<1 to 19
1 to 112
50 to 120
2500 to 2790
NR
NR
NR
NR
5 to 38
0 to 21
0 to 13
NR
NR
 Source:   Towill et al.,  1978
NR = Not  recorded
                                         3-25

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                                   TABLE  3-11
                 Chromium Concentrations in U.S. Drinking Waters3
Water
Tap water, Dallas, TX
100 largest cities, U.S. (1962)
380 finished waters, U.S.
Concentration, in
Median
4b
0.4°
7.5b
g/J, or ppb
Range
1 to 20
0.2 to 35
1 to 29
  (1962-1967)
3834 tap waters, U.S.
  (1974-1975)°
83 Midwestern cities, U.S.6
115 Canadian municipalities
  (1976-1977T
1.8
NR

<2.0
 0.4 to 8g
<5.0 to 17.0

 <2.0 to 4.1
 Source:  NAS, 1977
 Average value;  sampling date unavailable
Median value
 Greathouse and Craun,  1978
 iJ.S. EPA, 1975;  sampling date unavailable
f
 Mferanger et al., 1981
     of areas had detectable levels
NR = Not recorded
                                      3-26

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the U.S.  population were monitored  for metal  content  (Greathouse and  Craun,




1978).  The detection limit for Cr in  this  survey was 0.1  ppb.  The results of




this  survey,  presented in  Table 3-11, also  indicated that  28$  of  the  area



surveyed had Cr levels above the  detection  limit.   It should be mentioned that




the mean value of Cr in this study may be a little higher that reported since the




tap waters were not adequately flushed  before collection.




     In  two  surveys  (1976 to   1977)  of  115  municipalities  across  Canada




encompassing 63% of the Canadian population, the median and range of Cr concen-




trations were determined  (Meranger  et  al.f  1979,  1981).   The  median  and range




(within parentheses) of Cr concentrations in Canadian raw,  treated, and distri-




buted  waters  were  determined  to be  £2.0  (£2.0  to 5.0) ppb,  £2.0  (£2.0  to




4.0) ppb, and £2.0  (£2.0 to 4.0) ppb, respectively.








3.4.3.  Aquatic Suspended Materials and Sediments.  The concentration of chromium




in suspended materials  in several United States rivers was found to range between




37  and  460  ppm  on a dry weight basis  (Turekian  and Scott, 1967).   Chen et al.




(1974)  determined  the  concentration  in dry season  suspended  silts of Southern




California  waters  to be  =500 ppm in "natural" areas and  2000 ppm in urbanized




areas.



     Chromium concentrations determined for a variety of  bottom sediments are




shown  in Table 3-12.  For the areas  in the  United States  sampled,  sediment




chromium levels ranged from about 1 to 450 ppm.   An examination of Table 3-12




shows that  the chromium concentration in sediments from several Wisconsin lakes




and Southern Lake Michigan does not significantly decrease with depth.  A similar




finding  concerning  sediment chromium content  was made by  Bruland et al. (1974)




upon  the analysis  of chromium  levels in sediments of different depths from the




Southern California  basin.   These studies  of  sediment  chromium content versus
                                      3-27

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                                   TABLE 3-12

                     Concentration of Chromium in Sediments*
Chromium Concentration (ppm)
Area
Delaware Bay
New York City Bight:
Background
Sludge dumping area
Puget Sound
Houston ship channel , TX
Neches River, TX
Sabine River (low in
industrial activity)
TX-LA Border
Southern Lake Michigan:
surface sediments
sediments from >15 to
100 cm depth
estimated background
Illinois River
Non- industrial stream, IL
Buzzard Bay, MA
Wisconsin Lakes:
surface sediment
sediment from >50 cm depth
Median
NR
106
105
NR
NR
NR
NR
77
52
NR
17
6
33
NR
NR
Range
33 to 447
2 to 310
50 to 209
43 to 15*1
39 to 254
8 to 288
41 to 89
35 to 165
32 to 68
20 to 40
2 to 87
3 to 7
NR
1 to 49
0.8 to 35
•Source:  Towill et al.,  1978
NR = Not recorded
                                       3-28

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depth appear  to indicate  that  natural  background  chromium  levels  contribute




heavily to the chromium levels found in surface sediments.



     The  concentration  of  chromium  in  an  incinerated  sewage  sludge  ash was



determined to be 5280 ng/g.   The  concentrations  of metals in ash residue after



incineration is =4 times those present  in dried  sludge (Fraser and Lum, 1983).



Therefore,  sewage  sludges  are  expected  to  contain  high  concentrations  of




chromium.








3.4.4.  Soil.   The  concentration  of chromium in soil varies  in accordance with



soil origin.  Soils derived from nonserpentine areas can contain from traces to



300 ppm chromium.   Soils  derived  from serpentine areas can contain up  to  2% by



weight of chromium.  The chromium concentrations in selected  soils from various



parts of the U.S. are shown in Table 3-13.   Most  chromium in soils is apparently



insoluble.  Extraction  of  soils  with 2.5$ acetic  acid,  ammonium  acetate at pH



4.8, and  even with  0.1  N  HC1 have shown that only 0.01 to  4$ of the total soil




chromium is extractable (Towill et al.,  1978).








3.4.5.   Food.   The  chromium content  of  a variety  of foods  is  presented in



Table 3-14.  The values  given in Table 3-14 represent the average of several food



items  in each  category.    It can  be seen  from  this table  that  the  chromium



concentrations  in  different categories of food  determined  by Schroeder et al.



(1962) were lower than  the values determined by other  investigators.  It is not



known whether  the observed discrepancies are  due  to geographical and  seasonal



variations  in  trace  element content of foods or due  to  errors  in analytical



determinations.



     The  chromium content  in  acidic  foods is often higher than  other categories



of  foods.  The  values of Cr content  in a few commercial acidic foods which had
                                      3-29

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                                TABLE 3-13

            Chromium Content in Selected United States' Soils*
Location
     Soil
Characteristic
  Chromium content
     ppm or mg/g
Range           Median
Pennsylvania


Peninsular Florida

Florida

Missouri

New Jersey

Michigan
agricultural surface
and subsoil

surface and subsoil

surface and subsoil

on and off road soil

various soils

various surface soils
    NR            HI


 <1 to 1000       50

 <1 to 500        NR

    NR            71

 29 to 75         NR

3.2 to 17.6       NR
•Source:  Towill et al.,  1978
NR = Not recorded
                                 3-30

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                                   TABLE 3-14

                     Chromium Content in Various U.S. Foods
    Sample
Mean Concentration
  (ppm or [ig/g)
                                                         Reference
Fresh vegetables

Fresh vegetables

Fresh vegetables

Frozen vegetables

Canned vegetables

Fresh fruits

Fresh fruits

Fruits

Canned fruits

Dairy products

Whole fish

Meat and fish

Meat and fish

Sea foods

Sea foods

Grains and cereals

Grains and cereals

Fruit juices
      0.14

  0.03 to 0.05

      0.14

      0.23

      0.23

      0.09

      0.19

      0.02

      0.51

      0.10

  0.05 to 0.08

      0.23

      0.11

      0.12

      0.47

      0.01

      0.22

      0.09
Thomas et al., 1974

Schroeder et al.,  1962

Toepfer et al., 1973

Thomas et al., 1974

Thomas et al., 1974

Toepfer et al., 1973

Thomas et al., 1974

Schroeder et al.,  1962

Thomas et al., 1974

Schroeder et al.,  1962

Okuno et al., 1978

Toepfer et al., 1973

Schroeder et al.,  1962

Zook et al., 1976

Meranger and Soners, 1968

Schroeder et al.,  1962

Toepfer et al., 1973

Meranger, 1970
                                      3-31

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been in contact with stainless steel surfaces during harvesting, processing, or



preparation  for  market  are  shown  in  Table 3-15   (Stoewsand  et al.,  1979).



Jennings and  Howard  (1980) reported  slightly higher levels  of Cr  in British



commercial alcoholic  beverages  than the Cr  content in U.S. wines as  given in



Table 3-15.  The chromium contents in wines, beers, and spirits were reported to



be 0.45, 0.30, and 0.135  mg/£, respectively.   However, it is difficult to assess



the daily chromium intake in the United States from the Cr content in individual



foods.  The  chromium  contents in a selected  diet  composite were determined by



Kumpulainen et al.  (1979).  This study is the most recent diet study conducted



by FDA-USDA  (personal communication  with  Dr. W.R.  Wolf,  USDA).   The  chromium



intake  from  typical American  diets containing  43? fat was  determined  to be



62 + 28 |ig/day.  The corresponding intake from typical American diets containing



25$ fat was determined to be 89 ^ 56 |ig/day.








     3.4.5.1.  BIOCONCENTRATION IN FOOD CHAINS -- Several authors have found that




chromium concentrations  decrease  in higher trophic level  organisms  in aquatic



ecosystems  (Towill et al.,  1978).    For  example,  Mathis  and  Cummings  (1973)



detected =10 ppm chromium in worms, =5 ppm in clams,  =1.2 ppm in omnivorous fish,



and =1 ppm in carnivorous fish.  Lack of assimilation of chromium is probably the



major  reason that the organisms  of the higher trophic levels  contain lesser




amounts of chromium.



     A  bioconcentration  factor (BCF)  is the  ratio of the  concentration of  a



chemical in aquatic species to the concentration in the water in which they live.



An appropriate BCF can be used with data concerning food intake  to  calculate the



amount  of  chromium which  might  be  ingested  from  the consumption of  fish and




shellfish.   Residue  data  for a  variety of  inorganic  compounds  indicate  that



bioconcentration  factors  for  the  edible  portion of most aquatic  animals is
                                      3-32

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                                TABLE 3-15

     Concentration  of Chromium in a Few Commercial Grade Acidic Foods'
Commodity
Red cabbage
Red cabbage brine
Sauerkraut
Sauerkraut brine
Sauerkraut

Sauerkraut brine

Different brand
of honey
Vinegar
Hard cider
Cheese whey
Wine, Catawba
Wine, red concord
Wine, red concord
Container
glass
glass
can
can
flexible
pouch
flexible
pouch
glass

bulk
bulkb
bulk
lastiglass
( 1 year )
redwood
(8 months)
stainless
steel
(7 months )
Sample
PH
—
3.4
—
3.5
__

3.5

3.5 to 3-7

2.8
3.5
4.8
3.1
3.2
3.2
Cr Concentration,
ppm or jig/g (wet wt.)
2.6
10.3
0.3
0.3
0.2

0.2

0.04 to 0.18

0.01
0.004
0.01
0.02
0.07
0.02
fsource:   Stoewsand  et  al.,  1979
 Before acidification
                                    3-33

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similar,  except  that  for some  compounds  bivalve molluscs  (clams,  oysters,



scallops, and mussels) should be  considered a separate group.  An analysis (U.S.



EPA, 1980b) of data from a food survey was used to estimate that the per capita



consumption  of  freshwater  and  estuarine  fish  and  shellfish  is  6.5 g/day




(Stephan, 1980).  The per capita consumption of  bivalve molluscs is 0.8 g/day and




that of all other freshwater and estuarine fish and shellfish is 5.7 g/day.



     The BCF  for Cr(VI) in  fish muscle  appears  to be  less  than  1.0  (Buhler



et al., 1977; Fremm and Stokes,  1962), but values  of  125 and 192 were obtained



for oyster and blue mussel,  respectively (U.S. EPA, 1980c).  For Cr(III), BCF



values of  116, 153, and 86 were  obtained with  the American oyster (Shuster and



Pringle, 1969)t soft shell clam, and blue mussel  (Cappuzzo and Sasner, 1977),



respectively.  It  appears  that the two  valence states  of chromium(III  and VI)



have about the same BCF values, and  that the geometric mean of 130 can be used for



bivalve molluscs.  If the values of 0.5 (BCF for fish and mussels) and 130 (BCF



for bivalve molluscs) are used with  the  consumption data, the weighted average



bioconcentration factor for chromium in the edible portion of all freshwater and



estuarine aquatic organisms consumed by Americans can be calculated to be 16 on



the  basis  of  per  capita  consumption  of 0.8  g/day and  5.7 g/day  for  bivalve



molluscs, and fish and shellfish, respectively (U.S.  EPA, 1980b).








3.4.6.  Cigarettes.  Chromium has been determined  to be a  component of cigarette



tobacco.  Tobaccos  grown in the United States have been found to have a chromium



content ranging from 0.24 to 6.3 mg/kg (IARC,  1980).








3.5. INDICES OF EXPOSURE AND DOSE-RESPONSE RELATIONSHIPS



     Past exposure to low levels of chromium may be associated with higher than



normal levels of chromium in the hair.  Creason et al. (1975), however, reported
                                     3-34

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 that  this  association is tenuous  in  young children and women,  as  a result  of



 variation  in chromium  levels  related to  age and pregnancy.   Blood and  urine



 chromium  levels have  also  been  shown  to be  elevated following  exposure  to



 chromium,  although marked  variation  occurs  in  the  linearity between  exposure



 levels  and the  levels in body  fluids  as  a  result  of  sequestering and release  of




 chromium from body depots.  The only toxicologic index of exposure to chromium  is



 the development of perforated nasal septums.  This occurs only in the presence  of



 Cr(VI)  and is  associated  with  exposure to levels of 0.1 mg/nr*;  it  is not  known



 whether lower exposure will  also cause this disorder.








 3.5.1.   Chromium in Blood.  Chromium is absorbed through both  the  respiratory



 tract and gastrointestinal  system  (U.S.  EPA,  1978).   Exact values for  chromium



 absorption from the digestive  tract are  not known.  Cr(III) is poorly absorbed,



 whereas chromate is better absorbed (Mertz, 1969).



     In the  respiratory tract, water and  serum soluble chromates are  absorbed




 into the blood  system, whereas insoluble Cr(III)  particles and the inert oxides



 and hydroxides  of Cr(III) remain in lung tissue  (U.S. EPA, 1978).



     Once in the blood stream,  chromium compounds are bound  by  proteins (Gray and



 Sterling, 1950).   It  has  been  shown that ionic Cr(VI)  (injected intravenously)



 passes through the membrane of  red  blood  cells and binds to the globin moiety  of



 hemoglobin.   Once  inside  the  erythrocyte,   chromium  ions are  unable  to pass



 through the cell membrane and  move back  into  the plasma.   Hopkins and  Schwartz



 (1961!)  reported that,  in physiological  amounts,  cationic Cr(III)  is bound  to



 siderophilin and transported' to other tissues.



     Chromium disappears quickly from the blood  and is  taken up by other tissues



in the body, where it is  concentrated much more heavily (by a factor of 10 to 100)
                                     3-35

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than in the blood.  Therefore, blood levels of chromium are not a usable indica-



tor of chromium nutritional status (Mertz, 1969;  Mertz and Roginski, 1971).



     A wide range  of  values for chromium content  in blood has  been reported.



Schroeder et al. (1962) reported chromium levels in serum of 0.52 and 0.17 ppm,




whereas Doisey et  al.  (19&9,  1971)  found  a  chromium concentration  of  2  ppb in



serum.  Other chromium values reported have  ranged from 0.11 to 55 ppb in human



plasma, and from 5  to  54 ppb in red blood cells (Underwood, 1971).  Imbus et al.



(1963), working with United States subjects, found blood chromium levels ranging



from 13 to 55 ppb with a median of 27 ppb,  while Hamilton et al. (1973), studying



subjects from the United Kingdom, reported a blood level of 70 ppb chromium.



     However, researchers have discovered, through the use of new technology in




flameless AAS analyses, that the amount of chromium  in normal blood and urine is



in the range of 0.3 ppb, whereas previous measurements had  shown 0.73  to 150 ppb.



It was  considered  by  Guthrie et  al.  (1978),  Kayne  et al.  (1978)  and Anderson



(1981) that measurements  made before  1978 are probably  high as the result of




inadequate background correction for non-atomic absorption.  Thus, measurements



of chromium in normal body  fluids made  before  1978  are  probably useless (Toxic



Material News, 1982).



     Kayne et al. (1978) used flameless AAS with a tungsten halogen light source



for background correction  to determine the serum chromium levels in 8  normal male



subjects.  The tungsten halogen  light  greatly improved the background correction



in the near UV region where chromium is detected as compared to the standard Dp



light source.  With the elimination of non-atomic absorption, the mean level of



serum chromium was determined to be 0.1*1 \ig/l.  These results were in agreement



with those of Versieck et  al.  (1978)  in which serum chromium was determined by




neutron activation analysis.  Serum  obtained from 20  healthy subjects (duplicate



samples from  14  subjects)  had a range  of chromium  levels from 0.0382 to 0.351
                                     3-36

-------
     with a mean value of 0.16 |ig/fi,.   Using this data, the authors concluded that




normal human chromium levels in serum are in the sub-ppb range.








3.5.2.  Chromium in Urine.  A wide range of values for chromium content in urine




has been reported.  Hambidge (1971)  reported chromium levels in urine of 8.4 ppb




for adults and 5.5 ppb for  children  over a 24-hour period.  Imbus et al.  (1963)




reported median urinary concentrations of  chromium for adult males of 3-77 ng/£.




Renal  excretion  is the  major pathway  of chromium  elimination,  with  ^80$ of




injected chromium excreted in this manner  (Mertz, 1969).




     As discussed with chromium levels in  serum,  difficulties in determining the




low  levels  of chromium  in  urine were not  resolved  until  1978,  and presently,




normal  human levels  of chromium are considered  to be  in  the  sub-ppb range




(Anderson,  1981).   Using flameless  AAS with background correction by either a




tungsten  halogen  lamp or a continuum  source,  echelle,  wavelength modulated AA




system  (CEW-AA), normal  human  urinary chromium levels  were  determined to be >1




\ig/i  (Guthrie  et  al., 1978), >0.9  fig/A for most of 66 samples  (Kayne et al.,




1978),  and  between 0.05 to 0.58 \ig/i  for 48 males  and 28 females (Anderson et




al., 1982).  Veillon et al.  (1979) obtained excellent agreement between chromium




determinations of pooled urine samples using CEWM-AA  and chromium determinations




using stable isotope dilution methods measured by GC/MS.  The mean values  for the




respective methods  were  0.34  + 0.1  and 0.32 + 0.02  fig/Jl.   Increases in  urinary




chromium in humans receiving a daily supplement of 200 \ig Cr as CrCl- ranged from




mean levels  of  0.2 \ig/l prior to treatment  to 1.02  and 1.13 V-g/Si after  2 and 3




months.   This suggests  that  the high levels  of urinary  chromium  reported in




earlier studies were not likely to have resulted from changes in chromium intake.




Although Anderson (1981) cautions against  accepting absolute values for chromium
                                      3-37

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from earlier studies, comparisons and trends determined in a given study may be




valid.



     Franchini et al. (1975)  and Borghetti  et  al.  (1977) reporting about workers



exposed  to  chromium  in  the  chromium-plating industries  showed that  urinary



excretion and renal  clearance of diffusible chromium are  biological indices to



evaluate the  degree  of  current  exposure and  the  body burden  of the compound.



Franchini et  al.  (1978)  confirmed  their  earlier results  with an experimental



investigation using rats.



     Other authors have demonstrated a close relationship between the amount of



Cr(VI) in the air and urinary excretion  (Gylseth et al., 1977) or urinary excre-



tion of the metal corrected for creatinine  (Tola et al., 1977).  Gylseth et al.



(1977)  reported  in  an  abstract  that  welders  exposed to  a  concentration of



0.05 mg/m   (measured as  chromium)  had a urinary  chromium  concentration of



=40 ng/Jl, measured after work.



     Krishna et al.  (1976) studied 30 chrome workers who had nasal perforation.


                                                                      Q

The atmospheric concentration of chrome ranged  from  0.21  to 0.80 mg/m .  Urine



samples from the workers were collected at  the beginning of the  day's shift and



again  at  the end of the  day's   shift.   Before  exposure,  eight workers  had  a



concentration of chrome in the urine of j>0.20 ng/m£, whereas after exposure, 20



workers had such values.  In an  unexposed control group, there was no change in



the urine chrome values.   While  all  the workers tested had  nasal perforation, 66?



of them (20 of 30) had a urinary  chrome concentration of 2:0.20 jig/ai£»



     Mutti et al. (1979)  studied 22  welders  who worked with high chromium alloyed



electrodes.  The concentration of the breathing zone levels of chromium, deter-



mined  during the  1-month  exposure monitoring  period, ranged  from  0.017 to



          3                                                   3
1.000 mg/m  for  total chromium, and from  0.002  to  0.350  mg/m  as hydrosoluble



chromium.  Urine samples from the workers  were collected at the beginning and at
                                      3-38

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the end of the experiment.  These results suggest that the urinary concentration



of chromium at the end of a working period  is  affected  by recent exposure to the



compound.  At the same airborne concentration, the greater chromium body burden



is associated  with  greater excretion levels.  Even  after  a week following the




last  exposure, urinary  chromium  levels   provide  useful  indications  on  body




burden.



     Tandon  et al.  (1977)  have  reported  urinary  excretion of  chromium among




electroplaters and polishers in an  industrial  setting.   A  total of 12  subjects



were examined.  The range in duration of exposure to chromium in electroplating




processes ranged from 2 to 20 years  (mean duration for  12 workers = 11.1 years).



Chromium levels in workers' urine samples taken before  starting work ranged from



91 to  1116 \i&/l (mean value =  326.5 ng/£).   Urinary  chromium levels  did not



necessarily correlate to duration of exposure, but  a  slight  trend was  indicated.



Subjective complaints  included coughing, breathing difficulty,  dermal  itching,




depression, indigestion, body ache, and edema of the lower limbs.  Mean urinary




chromium  level  in  appropriate  controls was  38.1  \ig/i  (range:   0  to 78 \ig/SL).



Subjective complaints  did  not  appear  to correlate either with urinary  chromium



levels or with duration of exposure.  Urine samples collected at the  end of the



work  day,  however, would  perhaps  have provided  a  better  correlation between



exposure and complaints of illness.








3.5.3.   Chromium in Human Hair.    Schroeder  and  Nason  (1969)  reported a mean



chromium  concentration of  0.69 + 0.063 ppm for women.   Hambidge et al. (1972)



measured chromium concentrations at various distances from the hair root.  They



reported that  variation in the concentrations were due to  past fluctuations in



chromium  nutritional  status.   Hambidge and  Rodgerson  (1969)  reported higher



levels of chromium  in the hair of  nulliparous women (0.2 to 2.81  ppm) than in the
                                      3-39

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hair of parous women (0.04 to 1.14  ppm).  However, a later study by Hambidge and



Droegnueller  (197^) found changes in hair chromium levels due to pregnancy not to



be statistically significant.  Hambidge and Rodgerson  (1969) reported that hair



chromium levels in  3-  to 8-month-old infants were significantly higher  than in




those of 2-  to 3-year-old children.  Chromium  is obtained  through breast milk



during  nursing.   By the  second year  of  life, mean  chromium levels  in hair



approached values present in older humans.








3.6. SUMMARY








     Chromium  is  a metallic element  which,  when found in  nature,  is a stable



mixture of four separate  isotopes.  Inorganic chromium compounds occur in  valence



states ranging from -2 to +6; however, in the environment the Cr(III) and Cr(VI)



states are the most stable.  Chemically, the Cr(III) state is the most stable and



important form of inorganic  chromium  complexes.   Cr(VI)  compounds comprise the



most commercially important form of chromium, and they also appear to be the most




significant chromium compounds from an environmental standpoint.  Because Cr(VI)



is readily reduced in the presence  of organic material,  it is  rarely found in



nature apart from deposition by anthropogenic sources.



     Although chromite ore is not currently mined in the United States,  several



chromium chemicals are domestically produced  from imported  ores.   Sodium chro-



mate, sodium dichromate,  and chromic acid are three of the more important  commer-




cial chromium compounds   produced  in  the  United States.   Metallurgical  uses



constitute about 60$ of the largest market demand for  chromium.   Chemical uses



are the second largest  consumption  sector at 21$, followed by refractory  uses at



18$.
                                     3-40

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     Chromium emissions are released into the air, water, and land environments




from a variety of  industrial source categories including fossil fuel combustion,



cement production, incineration, cooling towers, refractory production, leather



tanneries,  steel  and  alloy  production,  electroplating,  and  chromite  ore



refining.   The largest chromium emission sources to the  air  are ore refining



plants, coal  combustion units,  refractory plants, and steel  and  alloy plants.



Principal sources of chromium  in  water systems  include  electroplating opera-



tions, leather  tanneries,  and  textile  manufacturing operations.   Significant



sources  of  chromium-containing  solid  wastes  that  are  land  disposed include



chromite ore refining operations and chromium chemical production plants.



     Recent monitoring of  the ambient air in many urban and rural areas of the



country has shown annual average chromium concentrations  to be in the range of



0.0052 to  0.1568  \ig/m .   The maximum concentration determined during any one



24-hour measurement  was about 2.48 ng/m .  The  chromium concentration in U.S.



waters varies  with  the type of surrounding  industrial  sources and the type of




underlying soils.  An analysis of 3834  tap waters in representative U.S. cities



showed a chromium concentration range of  from 0.4 to 8 ppb.  Chromium  levels in



soils vary with soil  origin and the degree of contamination from anthropogenic



chromium sources.   Tests  on domestic soils  have shown chromium  concentrations



ranging from  1  to 1000  ppm,  with the average concentration ranging from 14 to



about 70 ppm.
                                      3-41

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                           4.   SAMPLING AND ANALYSIS







     Chromium is ubiquitous in the environment and is an essential micronutrient




for man.  Therefore, the analysis of chromium encompasses a multitude of media,




namely, air,  water,  soil and sediments, foods, and a variety of biological solids



and fluids.  Numerous methods  are available for the determination of chromium in



these media.   This section  is  intended to be a general overview of the available



methods commonly used for chromium analysis.



     The analysis of chromium in a certain medium usually involves three distinct



steps, namely,  sampling and storage,  sample  pretreatment,  and analysis.   The



individual step,  as it pertains to the determination  of chromium  in  various




media, is discussed below.








1.1. SAMPLING AND STORAGE








1.1.1.  Air.








     4.1.1.1.   AMBIENT AIR  — Dusts  and  fumes of  chromium compounds  in the



ambient air are usually collected by high volume sampler at a flow rate of about



20 to  30 m^  hr~1  (Demuynck  et  al.f  1976).   Typical  filter media include cellu-



lose, polyethylene,  polystyrene, PVC, and glass-filter.  The suitability of the



filtering media  primarily  depends on their background impurity level, particle



retention  efficiency,  and  tendency  to become clogged.   Dams  et al.   (1972)



evaluated different  filter  materials and concluded that polyethylene filters are




most suitable for the collection of chromium particulates in atmospheric air.
                                      4-1

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     When size fractionation  is  required,  multistage cascade impactors  can  be




used for the collection of aerosols  (Broekaert  et  al.,  1982;  Winchester et  al.,




1981).  At a nominal sampling rate of  80 H min"  f  chromium  particles  of  aero-



dynamic diameter in the range of 0.04 to 25 |im have been separated by this method




(Broekaert et al.,  1982).




     Although no  specific information  is available, it is reasonable  to assume




that the filter paper protected from external contamination can be stored for  an




indefinite period prior to chromium  analysis.
     4.1.1.2.   OCCUPATIONAL AIR —  Chromium in occupational air is collected in




a way similar to ambient air.  A known  volume of air is drawn through a polyvinyl




chloride (PVC) or  cellulose filter  to collect chromium.  The  sampling rate is




maintained at  1.5 H min~   (NIOSH,  1977).   However,  it  has  been  determined by




Kneebone and Freiser  (1975)  that although the PVC filter can  be  stored for at




least 10 days, cellulose filters  cannot  be stored for more than  several hours




without some loss of Cr(VI).








     4.1.1.3.   STATIONARY SOURCE —  The sampling of chromium from stationary




sources, for example, stacks  of refuse incineration and  fossil  fuel burning




facilities, can be done by  collecting  the sample isokinetically (Block and Dams,




1976; Greenberg et  al.,  1978).  The sampling train  typically consists of a copper




probe, a rotameter, a manometer, and a pump with a capacity of 20 nr hr~ .  The




diameter of  the  nozzle is  adjusted to the stack  gas velocity to  achieve iso-




kinetic sampling conditions  (Block  and Dams,  1976).   Depending on gas tempera-




tures, glass-fiber, Teflon-fiber, and Whatman 41  filters have  been used (Block




and Dams, 1976; Greenberg et al., 1978).  However,  the use of glass-fiber filters




may result in a high blank value for chromium (Greenberg et al., 1978).
                                      4-2

-------
     For  studying distribution  of particle  size,  the  isokinetic  sampling is




typically   done   in  combination   with   a   multi-stage   cascade   impactor



(e.g., Andersen  cascade  impactor)  (Greenberg  et al.,  1978;  Block  and Dams,



1976).  The different stages of the cascade impactor can be used to determine the



cutoff aerodynamic diameters of the collected particles.



     The same method used for storing the filters from ambient air samples can be



used for the filters  collected from the sampling of stationary sources.








4.1.2.  Water.








     4.1.2.1.   DRINKING  WATER  —  To collect  distributed water  samples  from



household taps, the taps  are typically run  to waste at their maximum flow rate



for 5 minutes  to  clear  the  lines  of overnight standing water  (Meranger  et al.,



1981).  If suspended  solids are suspected in the water, filtration is typically



done on location  by  passing the  water samples through a 0.45  \an Millipore mem-




brane filter  (Meranger et al.,  1979).   In order to get representative samples,



some investigators  (Greathouse and  Craun,  1978) have collected monthly  samples



of finished water for a period of 1 year instead of one grab sample.  The water



samples are collected in  clean linear polyethylene screw-cap collection bottles.



Concentrated nitric acid  (1 mJl/100 m£ water sample) is added to each bottle and



they are filled to the brim to avoid  any air space.  The filled and capped  bottles



are transported in  heavy plastic  coolers  containing gel-type  freezer  packs.



Immediately upon receipt  in the laboratory, samples should be refrigerated at 4°C



(MSranger et al., 1981).



     Recently,  a  novel  method for  preconcentration  of  trace metals  in water



samples has  been proposed  by  Smits and  Grieken  (1981).   In this method,  a



cylindrical  plexiglass   filtration  unit  consisting   of   a  Nucleopore  0.4 urn
                                      4-3

-------
pore-size membrane  and  a 2,2'-diaminodiethylamine  (DEN)-cellulose filter  has
been used.  The Nucleopore filter was used to separate the suspended particles in
water and the DEN-cellulose filter allowed straightforward preconcentration of
trace cations by  a simple filtration step. The collection efficiency for Cr(III)
by this method has been claimed to be 90-100$.

     4.1.2.2.   RAW AND SURFACE  WATER — The sample  collection,  preservation,
transportation, and storage of raw and surface water samples are similar to those
for drinking water samples (Cranston and Murray,  1978;  Pankow et al.,  1977).  A
more representative sample can be obtained  by collecting  several  small samples
from different parts of the water body than by collecting one large sample at a
single point because of the inhomogeneity of the water body.

     4.1.2.3.   WASTEWATER — The sample collection,  preservation, transporta-
tion, and storage of  wastewater samples are similar  to those  for  other water
samples.   Grab samples  (Larochelle  and Johnson,  1978)  are often  collected but
wherever  a process  change is  suspected, a 24-hour  composite sampling  is  the
preferred or even necessary method.

4.1.3.  Soil  and Sediments.   Grab  samples  are typically  collected  for  the
analysis  of chromium  in soil  and sediment.   In  order  to  evaluate the chromium
content in  recent deposits,  samples  are collected from the upper few inches (2 to
3 inches) of sediment or  soil  (Pankow et al.,  1977).   The samples are usually
stored in polyethylene bags  or bottles  (Iwata et  al.,  1981; Pankow  et al., 1977).
The samples are  sieved through nylon sieves  (2 mm)  to remove gravel and leaves
(Iwata  et  al.,  1981).    The  collected samples  should  be   stored  at  4°C or

-------
refrigerated during  transportation and  storage to  minimize bacterial  action




(U.S. EPA, 1979).







M.I.4.  Food.  No specific information for sample collection and storage of food




samples could be found in the literature.  It is reasonable to assume that grab




samples collected in polyethylene bags or bottles and refrigerated during trans-




portation and storage should be an acceptable procedure for sample collection and




storage.








4.1.5.  Biological Samples.  Blood samples are typically drawn from donors with




10 mil vacutainers with disposable syringes and aluminum needles.  Heparin is used




as  the  anticoagulant.   Plasma samples can  be  obtained  by centrifuging freshly




drawn heparinized whole blood.  They  can be transported  at  4°C  in polyethylene




tubes and frozen to -20°C during storage. Urine should be collected from donors




in  polyethylene  bottles,  acidified,  and stored at 4°C  (Davidson  and Secrest,




1972).   Other  solid tissue samples  can be collected in  polyethylene bags and




transported at 4°C.  The tissue samples can be stored at -20°C.








4.2. SAMPLE PRETREATMENT








     Sample pretreatment is often  required  as  a technique to release the metal




from  the  sample matrix,  and  for  concentration and  separation from potential




interferences.   A  few  of the typical  pretreatment methods  have been discussed




below.  It should be emphasized that the pretreatment of samples is dictated by




the subsequent method of analysis, and in some  cases  the samples may not require




any pretreatment.
                                      4-5

-------
4.2.1.  Wet and Dry Ashing.   This pretreatment method is often used for chromium



analysis  in  air particulate,  biological samples,  foods,  soil,  and  sediment



samples in  combination  with atomic absorption, atomic  emission,  spectrophoto-



metric, and neutron activation analysis.  In  the wet digestion method, the sample



is digested with an acid or a mixture of acids depending on the sample matrix.  A




number of acid(s) mixtures including HNOg, H2S04 (NIOSH, 1977),  HN03/HF mixture



(Pankow et al., 1977), f^SO^/H^ mixture (Kumpulainen et al., 1979),  HNOg/H202



mixture  (Abu-Samra et al.,   1975),  HClO^/HgOg mixture  (Davidson and  Secrest,




1972), HNO-XHClOjj  mixture (Gallorini et al., 1976), HN03/H2S01}/  HCIO^ mixture



(Kuennen et al., 1982; Feldman et al., 1967) and HNC^/HjSOjj mixture (Bryson and



Goodall, 1981) have been used.  The two commonly used dry ashing procedures are



graphite  furnace  ashing  (Meranger  et al.,  1979;   Slavin,  1981)  and  low-



temperature oxygen plasma  ashing (Kumpulainen et al., 1979).  Some investigators



also  have  used a  combination of  dry  and wet  ashing  for  the  pretreatment of



samples (Kumpulainen et al.,  1979). Irrespective of the procedure, a particular




ashing method  is always optimized  to  minimize matrix interference and maximize



the chromium yield from the sample.








4.2.2.  Precipitation.  The direct precipitation of chromium from aqueous solu-



tion  by  such  reagents  as hydroxyquinoline  and  tannic  acid is  generally not



suitable  for  environmental  samples   containing  low  chromium  concentration.



However, the method of co-precipitation of chromium has been  successfully used in




recent  years.    In  this  method,  Cr(III)  is co-precipitated with Fe(III)  as



hydroxide at a pH of 8-8.5 (Cranston and Murray,  1978;  Pik  et al.,  1981).  The



co-precipitation of Cr(VI) has been accomplished by reducing Cr(VI) to Cr(III) by




the addition of Fe(II) and subsequent co-precipitation of hydroxides of Cr(III)



with Fe(II) at pH 8 (Cranston and Murray, 1978),  or by co-precipitating Cr(VI) by
                                      4-6

-------
Co(II)  and  ammonium  pyrrolidinedithiocarbamate  (APDC)  addition  (Pik  et al.,




1981).  The co-precipitated chromium is filtered from the solution and analyzed



by x-ray fluorescence or by flameless atomic absorption spectrometry.







4.2.3.  Solvent Extraction.  In this procedure, Cr(VI) is complexed with APDC at




a pH of 1.8-3.0 and extracted with an organic solvent, typically methyl isobutyl



ketone  (MIBK).   If Cr(III) is to be  extracted,  it must first  be oxidized to



Cr(VI) with reagents such as silver nitrate and potassium peroxydisulfate or with



potassium  permanganate  and sodium  azide  (Towill  et al.,  1978).   In  the gas



chromatographic  procedure,  chromium  is  chelated  with  1,1,1-trifluoro-2,4-



pentanedione or  trifluoracetylacetone  (HFTA) and  the chelate is extracted into



benzene.  This  method  has been used by a number of investigators (Lovett and Lee,



1976; Gosink,  1976).







4.2.4.  Chromatographic Method.  A number of chromatographic materials including




alumina, cation-exchange resin, anion-exchange resin, and chelating ion-exchange




resin have been used for the cleanup of impurities from chromium samples.  In the



anion-exchange  procedure,  the  solution  containing Cr(VI)  is  allowed  to  pass



through the resin bed at an optimum pH.  The Cr(VI)  retained on the resin bed is



subsequently eluted with a suitable eluent. The major drawback of this procedure



is that the  recovery  of Cr(VI) is often  poor.  The problem,  however, has been



alleviated by using an ascending flow technique or by in situ reduction of Cr(VI)



by Fe(II)  (Pankow  and  Janauer, 1974).   Marino and Ingle  (1981)  used medium



strength anion-exchange resin  to obtain  a satisfactory  recovery  of Cr(VI) from



the resin bed.




     In the cation exchange procedure,  the resin is used to retain the cationic



impurities while anionic Cr(VI) passed through the resin bed.  This procedure has
                                      4-7

-------
 been used  by Kneebone and Freiser (1975) for the analysis of Cr in occupational
 samples.
     The  chelatory  ion-exchange  procedure  was  employed by Leyden et al.  (1972)
 to absorb  quantitatively Cr(III)  from  a buffered solution.   By this procedure,
 Cr(VI) was detected by reducing it to Cr(IIl) with the addition of sodium  bisul-
 fite.
     The alumina procedure has been used for the separation of both Cr(III) and
 Cr(VI).  Larochelle  and Johnson (1978) adsorbed Cr(VI) on alumina column and used
 HC1 for the subsequent elution of Cr(VI).  Wolf et al. (1972), on the other hand,
 used an alumina column for the separation of Cr(III).   In this procedure, Cr(IIl)
 is precipitated with other cations by the addition of  excess 8-hydroxyquinoline.
 The dried precipitate  is dissolved in chloroform,  diluted with an equal volume of
 benzene, and  passed  through  an activated alumina  column.   Chromium was  eluted
 with a mixture of chloroform and benzene.

 4.3. METHODS OF ANALYSIS

     Chromium  can  be determined  by a  variety  of analytical  methods.    A  few
 analytical methods used for the determination of Cr are given in Table 4-1.   It
 should be emphasized that the detection limit and the percent-relative standard
 deviation (% coefficient of variation)  values given in Table 4-1 should be taken
 as  values   representative  of   the  specific  pretreatment  techniques  and
 instrumental methods  used  rather than  definite  data.  For example, both  the
 detection limit and percent CV values  obtained by  the same  instrumental method
may vary several-fold  depending on the  extent  of  preconcentration of the sample
and the method used for eliminating  interferences.   These values  may also vary
 considerably from laboratory  to laboratory  and  even within the same laboratory.
                                      4-8

-------
Catalytic method
                                                                       TABLE 1-1

                                                  Analytical  Methods  for  the Determination of Chromium
Method
Graphite furnace AA


Flame AA


Spectrophotometrio
Type of Sample
blood, urine, and other
biologic samples3
natural waters
raw, treated, and dis-
tributed water0
raw, treated, and dis-
tributed water0
natural waterd
blood, urine, and other
biologic samples6
tissue samples^
Preconcentration
none
co-precipitation
with Fe(OH)
none
APDC-MIBK
extraction
APDC-MIBK
extraction
an ion exchange
MIBK extraction
APDC-MIBK
extraction
Selectivity
total Cr
Cr(III); Cr(VI)
can be reduced
to Cr(III) by
Fe(II)
total Cr
Cr(Vl); Cr(III)
can be oxidized
to Cr(VI)
Cr(VI); Cr(III)
can be oxidized
to Cr(VI)
Cr(VI)
Cr(VI)
Cr(VI); Cr(III)
can be oxidized
Detection
Limit
0.1 ug/l
0.001 ug/l
2 ug/l
0.6 ug/l
0.05 ug/l
0.1 ug/l
10 ug/l
1.2 ug/l
* CV
(at sample
concentration)
5.2*
(10 ug/l)
5*
(0.14 ug/l)
15* (2 ug/l)
20* (2 ug/l)
5* (3 ug/l)
20* (0.1 ug/l)
15* (25 ug/l)
0.4* (800 ug/l)
Interference

Matrix interference
can be avoided by
wet ashing.
none reported
none reported
none reported
none reported
none reported
none reported
Fe, Hi and PO^
may interfere





3-
                         air particulates"
Neutron activation       air psrtloulatesh


                         freshwater^
                                                      none
                                                      none
                                                      none
to Cr(VI)

Cr(VI)
                                                                           total Cr
                                                                           total Cr
1 ng          6* (<100 ug/l)  Pb,  Cu,  Cr(III),
                              Fe(III), and V(?)
                              may  interfere

30 ng/n            15*        none reported
              (1.3 ng/m3)

0.12 ug/l     21* (1.4 ug/l)  none reported

-------
                                                                      TABLE M-1  (oont.)

                                                     Analytical Methods for the Determination of Chromium
Method
Gas chromatography
(electron capture
detection)
Gas chromatography
(AAS detection)
Gas chromatography
(MS detection)
Liquid chromatography
(coulometric detection)
X-ray fluorescence
Type of Sample
natural waters
blood, serum, orchard
leaves
bloodm
natural water11
dried solution deposit0
Preconcentration
HTFA-benzene
extraction
HTFA-benzene
extraction
HTFA-benzene
extraction
1 i sample concen-
trated to 10 mi
none
Selectivity
Cr(III) and
Cr(III)
Cr(III)
Cr(VI)
total Cr
Detection
Limit
Cr(VI) 0.1 ug/l
<1 ng
0.5 pg
0.8 ug/l
1.5 ug/gP
* CV
(at sample
concentration)
2.6* (1.9 ug/l)
<6* (1 ng)
9* (10 ng/g)
<2* (90 ug/l)
'" ?.
Interference
none reported
none reported
none reported
SO,.'2, PO.3'
may inerflre
absorption by
   (energy dispersive)
j_ X-ray fluorescence
I  (energy dispersive)
   Differential pulse
   polarography
   Emission spectroscopy
   (inductively coupled
   plasma source)

   Mass spectrometry
surface water and drinking
waterq
natural waters
natural waters
variety of samples
chelating ion-
exchange membrane
                              none
                              none
                              none
Cr(III)
                     Cr(VI)
                     total Cr
                                                   total Cr
              (1 jig/cm*1)      matrix and dif-
                              ference in
                              particle size
                              may cause error

0.8 ug/l      10-15*          excess alkali and
              (1 ug/l)        alkaline earth
                              metals may inter-
                              fere

10 ug/l       34* (61 ug/l)   excess Cu(II) and
                              Fe(III) may inter-
                              fere

1.8-6         05*             none reported
                                        0.05-1 ug     20* (photo-     Any species having
                                                      graphic)        the sane m/e ratio
                                                      3* (electrical) as Cr nuclide may
                                                      0.5* (isotope   interfere
                                                      dilution)

-------
   Chemiluminescence
   (lophine)
                                                                      TABLE  1-1  (cont.)

                                                     Analytical Methods for the Determination of Chromium
Method
Chemil uminescence
Detection
Type of Sample Preconoentration Selectivity Limit
natural waters and orchard none Cr(III) 0.02 (ig/l
leaves1*
% CV
(at sample
concentration)
201 (at 2.3
ppm)
Interference
Fe(III), Fe(II),
Co(II), S032'.
and NO-" may
Interfere
                            natural waters'
        ion exchange resin   Cr(VI)
0.015
Pe(III), Fe(II),
Co(II), Cr(III),
Mnott", and
other cations and
anions may interfere
   Davidson and Secrest, 1972

£r  Cranston and Murray, 1978

Z? °M*ranger et al.f 1981

   Sankow and Janauer,  1974

   Veldman et al., 1967

   fBryson and Goodall,  1981

   ^Kneebone and Freiser, 1975

   T)emuynok et al., 1976

   1Bhagat et al.,  1971

   •'salbu et al.,  1975

   T,ovett and Lee, 1976

         , 1976
"Volf et al., 1972

"uarochelle and Johnson, 1978

°Camp et al., 1975

pKuhn, 1973

fyanGrieken et al., 1977

rCrosmun and Mueller, 1975

8Quinby-Hunt, 1978

 Boumanns and deBoer, 1972

uAhearn, 1972

VElser, 1976

WHoyt and Ingle,  1976

*Marino and Ingle, 1981

-------
The following sections briefly describe some of the methods used for the analysis



of chromium.








1.3.1.  Atomic Absorption Spectrometry  (flame).   This method has  been used by




various  investigators  for the  determination  of  Cr  in  surface water,  sewage



effluent, and biological samples.  The determination of Cr by the air-acetylene



flame is prone to interference by other elements (Thompson and Wagstaff, 1979).



This  problem  can be avoided  by using  nitrous oxide-acetylene  flame  but  this



results  in  a  decrease  in the  detection limit  (Thompson and Wagstaff,  1979).



Therefore, most  investigators have used  air-acetylene flame for the determina-



tion of Cr.  However, for samples with low Cr concentration,  pretreatment of the



samples  providing preconcentration of  Cr and the reduction  of  the interfering



effects  from  other  ions are required.   Thompson and  Wagstaff  (1979)  used an



evaporative technique on  a  hot plate  to concentrate the sample 5-fold.   A 2%



ammonium perchlorate solution  (W/V) was used to suppress the interelement inter-




ference  effects.    Better   methods  for  pretreatment  of  samples  include



ion-exchange separation (Pankow et al.,  1977;  Pankow and  Janauer,  1974),  APDC-




MIBK  extraction  (Gilbert and  Clay,  1973) and MIBK  extraction  (Feldman et al.,



1967).  The  flame AAS technique is not a preferable method for samples with very



low Cr  concentration since this  method has a  much  higher detection limit than



flameless AAS (Slavin,  1981).








4.3.2.  Atomic  Absorption  Spectrometry   (flameless).    In  the flameless  AAS



method, the sample is atomized directly  in a graphite furnace,  carbon rod, or



tantalum filament instead of  a flame.  It is one of the most attractive methods




for the analysis  of solid and liquid biological  samples since the method does not



generally require sample preparation. Despite  the high sensitivity of the flame
                                     4-12

-------
less AAS method, it may suffer from certain disadvantages.   For example,  matrix
interference, background or  nonspecific absorption effects,  volatilization of
some Cr during dry ashing, and adsorption of Cr on the walls of crucibles  during
dry ashing can  cause error in  analysis.  The matrix interference effect  in the
case  of water  samples  can  be  overcome by  chelation  and solvent  extraction
(Meranger et al., 1979,  1981) or  by the co-precipitation technique (Cranston and
Murray, 1978), and in the case of biological samples, by wet disgestion (Davidson
and Secrest, 1972) prior to introduction of the sample into  the graphite furnace.
Background correction for non-specific  absorbance  can be made with a deuterium
arc corrector  (Cranston and  Murray, 1978) or  better  yet,  by continuum source,
echelle,  wavelength-modulated,   atomic  absorption  spectrophotometer (CEWM-AA)
(Kumpulainen  et al.,  1979).   The  volatilization  of  Cr  during dry  ashing at
temperatures  of 700°C or higher,  particularly during the  analysis  of complex
matrices  such  as  food, may  be substantial  (Kumpulainen  et al.,  1979).   Low
temperature  ashing,  for example, with oxygen  plasma  at  150°C,  would eliminate
this  problem and  the  problem of Cr adsorption on crucible walls.  However, the
low temperature ashing  may not  be  suitable for some biological samples (bovine
liver),  and  some biological  materials such  as brewer's  yeast may contain acid
insoluble material that can strongly adsorb chromium (Kumpulainen et  al., 1979).
However, these  problems have been overcome by utilizing dry ashing at 500°C with
sulfuric acid and  hydrogen peroxide as ashing  acids (Kumpulainen et  al., 1979).
The problems of sorption and volatilization can also be eliminated by the use of
"platform" technique described by  Slavin (1981).
      Versieck  et  al.  (1978)  reported that  chromium levels in serum  from normal
subjects was 0.16 jig/i, when determined by carefully conducted  neutron activation
analysis.   In  a  survey of  previous  reports  of chromium  levels  determined by
flameless AAS, the range of values was 0.73 to  150 ng/&, and the  levels tended to
                                      4-13

-------
become  lower  as  improvements  in  detection limits  occurred.   These  results



prompted an intensive study of possible sources of artifacts  in the flameless AAS



analysis of biological fluids by Kayne et al.  (1978)  and Guthrie et al.  (1978).



Guthrie et al. (1978), in a study of  the  effects of purge gas, char temperature,



sample  volume,  and  graphite tubes on  the determination of  low ppb  levels  of



chromium, demonstrated  that all  these parameters affected  the  background non-



atomic absorption, and  that there  was a  direct correlation  between background



absorption and  the apparent chromium content  of the  sample.   It was suggested



that the background  correction was inadequate as the result of the low intensity




of the D  lamp at  the  near UV wavelength used in chromium analysis.  Kayne et al.



(1978) came to a  similar conclusion, and modified the background correction by



replacing the usual  D  lamp with  a  tungsten  halogen lamp.   This  modification



provided adequate background correction, and  serum chromium  levels determined




for 8 normal male subjects  averaged  0.14 \ig/H,  while  most  measurements  from 66



randomly chosen urine samples were =0.9  ng/&.  Guthrie  et al. (1978), Kayne et



al. (1979),  and  Anderson (1981) warned that earlier reports of  chromium levels in



urine  and  serum  should be considered  artificially  high  in  light of  these




findings.
4.3.3.  Emission Spectroscopy.   In  emission  spectroscopy,  prepared samples are



excited with a flame,  arc, spark, or plasma and the resulting light is dispersed



with a monochromator.   The characteristic emission lines of the excited elements



are recorded electronically or on a photographic plate.   Because  of its better



sensitivity, inductively compiled plasma atomic emission spectrometry (ICP-AES)



has been more extensively used in recent  years than the other modes of excitation



(Towill et al., 1978; Slavin, 1981).  Since  the  sample  is generally introduced
                                      4-14

-------
into the  plasma source  by pneumatic  nebulization, all  solid and  biological



samples require wet digestion prior to analysis.  The wet digestion also tends to




minimize the matrix effects.  A HF/HN03 digestion at 170°C has been employed for



the simultaneous multi-element  analysis of air  particulate  matters  (Broekaert



et al.,   1982).     Similarly,   wet   digestion   by   direct  chelation   with




1,1,1-trifluoro-2,4-pentaedione (also known as trifluoroacelytacetone) (HTFA) or



hexafluoroacetylacetone (HHFA)  has been used for the cleanup and volatilization



of Cr in biological samples (Black and Browner, 1981) prior to ICP-AES analysis.



Recently, a direct  atomization technique for raw agricultural  crop samples has



been presented (Kuennen et al., 1982).  The technique employs a 30-minute pres-



sure dissolution of sample composite with 6M HC1 at 80°C in linear polyethylene



bottles.  The dissolved samples after proper filtration  by Teflon filters can be



aspirated directly  into  the  plasma  source.  Combined with  real  sample matrix



calibration technique, it has been suggested that this method provide comparable



recovery and precision as  obtained by  the  more  time-consuming  conventional wet




ashing methods.








4.3.4.  Neutron Activation Analysis.  Neutron activation analysis is one of the




most  sensitive modern analytical techniques for   the  determination of  trace



elements.  Neutron activation analyses are applicable to many kinds of environ-



mental samples including air particulates,  dusts, soils, fresh and marine water,



sediments,  biological  liquids  and solids,  and  foods.   The samples  are  often



irradiated without prior chemical treatment.  A detection limit of 0.12 ng/& in




freshwater  (Salbu   et al.,  1975)  and  0.2 jig/g  in  biologic materials  (Spyrou



et al., 1974)  has  been  reported for samples analyzed without  chemical  proces-



sing.   Lower  detection  limits generally  can  be achieved  if  the samples  are



chemically processed to separate and concentrate chromium.   For example, in the




chemical processing of samples  typically by ion-exchange  separation,  detection
                                     4-15

-------
limits of 0.1 ng/fc for river water,  3 ng/& for seawater, and in the ppb range for


biological  and  environmental  samples  have  been  reported   (McClendon,  1974;


Robertson and Carpenter, 1974).


     One distinct advantage of this method is the reduced problems arising from


reagent contamination.  Even if  chemical  processing is required, post-irradia-


tion contamination is of no consequence in introducing error to the final result„


In addition to the problem of acquiring a neutron source, this method has another

                                                                          ph
disadvantage.  Due  to the intense  x-ray  or  bremsstrahlung activity from   Na,


3 Cl,   K, 5 Mn,  and ^ p in many  samples, the irradiated samples usually must be


cooled for several weeks before measuring  Cr concentration.  However, chemically


separating  the offending  nucleides can  reduce  the cooling  period  to  about


24 hours (McClendon, 1974).





4.3.5.  X-ray Fluorescence.  The x-ray fluorescence method is commonly applied


to solid samples.  In this  technique, the sample  is bombarded with high energy


photons, for example, low energy  x-ray or gamma photons or  with particles such as


protons.  The intensity of the characteristic emitted x-ray is currently measured


by two techniques, namely, energy-dispersive analysis and wavelength-dispersive


analysis.  The resolution of wavelength-dispersive analysis is much better than


energy-dispersive analysis  which often  permits the  former method to determine


the oxidation states of an element (Quinby-Hunt,  1978).


     This technique is useful  for trace analysis  in solid samples, namely,  air


particulates  (collected  on filters),  sediments,  biological  specimens,  and


filtered suspended  solids  from aqueous media.   However,  for  aqueous samples,


sample preparation leading  to the deposition  of the element on a filtering medium


is required.  This deposition can be done  by a variety of  methods.  The obvious


and simple method  is the evaporative  drying of the sample on a Mylar film (Tanaka
                                     4-16

-------
et al.f 1981).  Other methods of sample deposition include retention on chelating



filters  (Smits and  Van Grieken, 1981),  vapor  filtration  technique (Greathouse




and Craun,  1978) and co-precipitation (Pik et al.,  1981).  The last three methods



not only allow deposition of Cr on  a filtering media, but also permit separation



and concentration of Cr from the aqueous phase during the deposition step.



     Sample preparation in the case of. solid samples is an important requirement



for this method.   In order to avoid absorption and scattering by sample matrix,



the size and shape of  the particles on the films should be controlled by pressing




the deposits into thin wafers (Towill et al., 1978).








4.3.6.  Colorimetric.  The usual colorimetric method for the determination of Cr



involves oxidation of Cr(III) to Cr(VI), followed by complexation with diphenyl-



carbazide.   However,  the sensitivity of  the method is such  that  samples with



relatively high Cr concentrations can  be determined fay this method.  This method



has been proposed by  NIOSH  (1977)  for the  determination  of Cr in occupational




atmosphere.  However, the NIOSH method of digesting the filter with 0.5 N H^O^



for the determination of Cr(VI) may not be suitable for use with welding fumes.



A carbonate leaching method  has been  described for the determination  of Cr in



welding fumes and other complex matrices (Thomsen and Stern, 1979).  Bryson and



Goodall  (1981)  have  described a  modified  diphenylcarbazide  spectrophometric



method for the determination of Cr in biological tissues.
4.3-7.  Gas Chromatography. .



     The gas chromatographic method is  suitable  for a variety of environmental




samples. The analysis of water samples by this method  does not generally require



pretreatment before complexation (Lovett and Lee, 1976).  Samples  that are solid,
                                     4-17

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namely, particulate matter, and biological samples are first digested to get Cr




in solution.  The Cr is  then  quantitatively  chelated with 1,1,1-trifluoro-2,4-
                                                  »



pentanedione  (HFTA)  or  hexafluoroacetylacetone  (HHFA)   to form  a  thermally



Stable, volatile Cr(III)  complex.  To determine Cr(VI), it should be reduced to




Cr(III)  by a reducing  agent, such  as sodium  sulfite.    The  Cr(IlI)-HFTA  or




Cr(III)-HHFA complex is  extracted  into an organic solvent, usually  benzene or




hexane,  and  an  aliquot  is injected into the GC  column.   Recently a  direct




chelation  (without pretreatment)  method has  been   described  for  biological




samples  (Black and Browner, 1981).




     The detection of Cr(III)  complex can  be  accomplished  by a variety of detec-




tors.  The sensitivity  of this  method  is dependent  on  the detector used.   A




number of  detectors including electron capture (Lovett and Lee,  1976),  atomic




absorption  (Wolf,  1976),  mass spectrometric  (Wolf et al., 1972),  and  ICP-AES




(Black and Browner, 1981) have been used.




     The gas  chromatography with  any  of  the  detection  methods described  has




excellent sensitivity for Cr determination, but the mass spectrometric method is




extraordinarily sensitive and specific.








4.3-8.   Chemiluminescence.  Luminal (5-amino-2,3-dihydrophthalazine-1,4-dione)



and  lophine  (2,4,5-triphenylimidazole)  emits light  when oxidized  by hydrogen




peroxide.  The first oxidation in basic  solution is catalyzed by Cr(III) and the




second by Cr(VI).  The design  of different Chemiluminescence instruments for the




determination of Cr vary primarily in  the technique  for  mixing  the sample and




reagents.  The three types of sample modules which are commonly used are discrete




sampling system, flow system, and  centrifugal analyzer (Hoyt and Ingle,  1976).




All these sample modules  have been used for  the analysis  of Cr  (Towill  et al.,




1978).
                                     4-18

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     The chemiluminescence method has been  used primarily for the analysis of Cr




in water  and  in biological samples  (Marino and Ingle,  1981;  Hoyt  and  Ingle,



1976).  The method is fast, economical, and has a high sensitivity.  The sensi-



tivity can be further increased  by preconcentration  of  the samples (Marino and




Ingle, 1981).  However, this method  has  not been as widely used as some of the




other methods.








4.3.9.  Polarography.  Two recent variations of the method, namely, single-sweep




polarography and differential  pulse  polarography,  have both been  used for the



detection of Cr in natural  waters (Whitnack, 1975;  Crosmun and Mueller, 1975).



The  polarographic  method  has  a comparatively  lower  sensitivity  than  other



methods and is not currently popular for Cr analysis.








4.3.10. Mass  Spectrometry.   The spark-source  mass  spectrometric  method  is



applicable to virtually any matrix,  but the results  are  only  semiquantitative




(Towill et al.,  1978).  The precision and accuracy of  this  method can be greatly



improved by using the isotope dilution technique.  This method has been used to



establish the Cr  content  in NBS  bovine  liver  (Dunstan  and Garner, 1977).   The



mass  spectrometric  method  is  rarely  used  for  the  routine  determination  of



chromium.








1.3.11. Catalytic Method.   This  method has  been used for  the  determination of



Cr(VI) in occupational samples (Kneebone and Freiser,  1975). In this method, the



microdetermination of Cr(VI) is done by  means of  Cr catalyzed  oxidation  of



o-dianisidine by hydrogen peroxide.    The  reaction  rate is  monitored  spectro-



photometrically.  This  method has  limited  application  (occupational  samples)
                                     4-19

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because of  interferences  by various  cations,  and it  is rarely  used  for  the




determination of Cr.







4.3.12. Liquid Chromatography.  This  method in combination with  a  coulometric




detection has been used for the determination of Cr in water samples  (Larochelle




and Johnson, 1978).  In this method Cr(VI) is separated by an alumina column and



the separated solution is passed  through a flow-through coulometric detector.



Although the method has been purported to have good sensitivity and precision, it



has found little application  in the  analysis of chromium.








4.4. CONSIDERATIONS IN ANALYSIS








     The determination of  Cr in samples containing trace amounts of the  compound



requires special precautionary measures from the initial  sample collection step



to the final analytical manipulations of the samples.  For example,  contamination




of samples during collection  should  be avoided  by the use of Cr-free equipment.



This is particularly true  for the  collection of biological  samples.   Biological



tissue samples are sometimes collected by hospital personnel who  use stainless



steel scalpels,  trays, utensils, etc.  containing about  18? Cr.  This may produce



unacceptable contamination in the  samples.   Similarly,  sample containers should



be free from the possibility of sample contamination.  Polyethylene bottles and



bags are  particularly suitable as sample  containers.   Even with polyethylene



bottles, adsorption of Cr  on  the surface  of  the  container from water samples may



be a  serious problem.  Therefore,  acidification of aqueous  samples  prior to



storage is necessary to avoid adsorption losses.



     The possibility of sample contamination and losses  during analytical pre-



treatment of the samples  should  be  avoided.    Care  should be taken that only
                                     4-20

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reagents of the highest purity are used.   Even so,  the quantity added should  be




limited to a minimum to avoid  unnecessary  buildup  of  contaminants.   The use  of




Cr-containing grinding or homogenizing equipment can introduce Cr-containination



into the samples.   Grinding such samples with an  agate mortar  and  pestle  is a




better procedure.   The analysis  of  Cr,  especially  in biological samples,  is




complicated by extreme matrix effects, possible volatility of some Cr complexes,




and the inherent property of  Cr-complexes  to  bind  non-specifically  to reaction




vessels, graphite tubes, or  other equipment.  The methods used to minimize these




problems have been discussed in section 4.3-2.




     The problem of developing accurate  data  from  Cr  analysis,  particularly  in




food and biological samples, is amply illustrated by the large variations in the




interlaboratory comparison  data  (Towill  et al.,  1978).  The  problem is further




aggravated by the non-availability of Standard Reference Materials (SRM).  Only




recently has  the National  Bureau of Standards  (NBS) issued the Cr certified




materials, such as brewer's  yeast  (SRM-1569),  bovine  liver  (SRM-1577),  orchard




leaves  (SRM-1571),  spinach leaves  (SRM-1570),  pine  needles  (SRM-1575), and




tomato leaves  (SRM-1573).   In  view of the absence of  suitable  comparison  with




SRMs, the older data should be interpreted with skepticism.




     Another problem in dealing with the analytical methods  is their ability  to




distinguish between Cr(III)  and  Cr(VI).   This is  particularly  important since




Cr(VI) has been associated with health hazards,  while Cr(III) is of substantially




less concern  and  is in fact necessary for the maintenance of a normal glucose




tolerance factor (Marino and Ingle,  1981).  The problem for the determination  of




the two oxidation states of Cr is not  critical in  foods,  sediments,  soils, and




biological samples,  because  Cr is genrally present in the Cr(III) state in these




samples.   However,  Cr may  be  present  in both oxidation states  in  ambient and




occupational air,  and in water samples.  It should also be recognized that Cr(VI)
                                     4-21

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may be present in both water soluble and water insoluble form in these samples.




It  is  necessary to  distinguish  these two  forms  of Cr(VI),  as they may  have




different genotoxic properties (Thomsen and Stern, 1979).  Some of  the  methods



that distinguish between Cr(VI)  and Cr(III) have already been  discussed.   The




analytical method for the determination of soluble and insoluble forms of Cr(VI)




in occupational air samples has been described by Thomsen and Stern  (1979).








4.5. COMPARISON OF METHODS








     Based on versatility,  sensitivity,  and precision,  the three most important




methods that have found wide application are graphite furnace AAS, x-ray fluore-




scence,  and neutron  activation  analysis.    Of these  three methods,  graphite




furnace AAS has  the advantage of being the least expensive method.  The disadvan-




tage of graphite furnace AAS  is  that  it  cannot  be used for  simultaneous multi-




element determinations.  Both neutron activation and  x-ray fluorescence, on the




other hand, are commonly used for simultaneous multi-element analysis.  X-ray




fluorescence has the advantage over  both neutron activation and graphite furnace




AAS analysis in that it can differentiate between the various oxidation states of




Cr without prior pretreatment  of  the samples.  The disadvantage of x-ray fluore-




scence is that it often requires time-consuming pretreatment  of the samples.  The




use of  neutron activation analysis  normally requires  about  a  2-week  cooling




period if post-irradation separations are not performed.  Thus, the technique is




not suited for on-line or  rapid analysis of chromium.




     The choice of a particular analytical method for Cr analysis is dictated by




several factors including the  type of sample to analyzed, concentration of Cr in




the sample, and the scope of  the analysis.   These factors,  in combination  with
                                     4-22

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others such  as  the desired precision  and accuracy and  the cost  of  analysis,



should be weighed in selecting a particular analytical method.
                                     4-23

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                   5.   CHROMIUM  METABOLISM  IN  MAN  AND  ANIMALS








5.1.  ROUTES OF CHROMIUM ABSORPTION








5.1.1.  Chromium Absorption and  Deposition  by  Inhalation.  An important route of




exposure  to  chromium compounds  is through inhalation of  chromium  containing




particles and aerosols.  Although there are no specific data on  the deposition of




chromium in the lungs, some general considerations of factors affecting deposi-




tion as presented  in U.S.  EPA  (1982) would  likely  also  apply to particles or




aerosols containing chromium.   During inhalation  (and exhalation)  a portion of




the  inhaled  aerosol  may be deposited  by  contact with  airway surfaces  or be




transferred to unexhaled  air.  The remainder is exhaled.  The portion transferred




to unexhaled air may be either deposited by contact with airway  surfaces or later




exhaled.  These phenomena are complicated by interactions that may occur between




the  particles,  other  gases such  as  endogenous  ammonia,  and  the water  vapor




present in the airways.




     The mechanisms involved in the deposition of inhaled aerosols are affected




by physical and chemical  properties, including  aerosol particle  size distribu-




tion,  density,  shape, surface  area,  electrostatic  charge, hygroscopicity or




deliquescence, chemical composition, gas diffusivity and solubility, and related




reactions.  The geometry  of the respiratory airways  from nose and mouth to the




lung parenchyma also influences aerosol deposition; the important morphological




parameters  include  the   diameters,  lengths,   inclinations to  vertical,  and




branching angles of airway segments.   These factors affect the extrapolation of




data  between  different   species  that  have  respiratory   tracts  of  different




geometry.    Physiological  factors that  affect  deposition  include  breathing
                                      5-1

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 patterns, air flow dynamics in the respiratory tract,  and  variations of  relative




 humidity  and temperature within  the  airways.  Clearance from the respiratory




 tract  depends on many factors,  including site of deposition, chemical  composi-




 tion  and  properties  of the deposited particles, reaction products, mucociliary




 transport  in the  tracheobronchial  tree, macrophage  phagocytosis in  the deep



 lung,  and pulmonary  lymph and blood flow.




     Aerodynamic  properties  of  aerosol  particles  depend  upon  a  variety  of




 physical  properties,  including  the size  and  shape of the  particles  and their




 physical  densities.   Two important aerodynamic properties of aerosol particles




 are the inertial properties, which are most important for particles larger than




 0.5  M-m in  diameter   and  are related to  the settling speed in  air  under the




 influence of the earth's gravity,  and  the diffusional  properties, which  are most




 important for particles  smaller than  0.5 |im in diameter and are related to the




 diffusion coefficient (Fuchs,  1961).   When particles are inhaled,  their aero-




 dynamic  properties,   combined  with  various  anatomical  and breathing  charac-




 teristics,  determine their  fractional  deposition  in various  regions  in  the




 respiratory tract.




     The  respiratory tract  includes  the passages of the nose, mouth,  nasal




 pharynx,  oral pharynx,  epiglottis, larynx,  trachea,  bronchi,  bronchioles  and




 small  ducts  and alveoli of  the pulmonary acini.   With  respect to respiratory




 tract  deposition  and clearance  of  inhaled  aerosols,  three  regions   can  be




 considered:  (1) extrathoracic,  the airways  extending  from the nares down to the




 epiglottis and larynx at the entrance to  the  trachea  (the mouth is  included in




 this region  during mouth breathing);  (2) tracheobronchial  region,  the primary




 conducting airways of the lung from the trachea to the terminal broncioles (i.e.,




 that portion of the lung respiratory tract having a ciliated epithelium); and 3)




pulmonary region, the parenchymal  airspaces  of the lung,  including the respira-
                                      5-2

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tory bronchioles, alveolar  ducts,  alveolar sacs,  atria, and alveoli (i.e., the
gas-exchange region) (Morrow et al., 1966).
     The  behavior of  inhaled  particles in  the  respiratory airways  and their
alternative  fate  of  either  deposition  or  exhalation   depend  upon  aerosol
mechanics under  the  given physiological and  anatomical condition (Yeh et al.,
1976; DuBois and  Rogers,  1968).   Contact  of  particles  with moist airway walls
results  in  attachment  and irreversible removal of  the  particles from the air-
stream.   The  contact process can occur  during inspiration  or  expiration of a
single breath  or subsequently if a particle  has  been transferred to unexhaled
lung air  (Engel et al., 1973; Davies, 1972; Altshuler,  1961).
     There  are five  primary physical  processes  that lead  to  aerosol particle
contact with the wall of the airways.  Gravitational settling occurs because of
the influence  of the earth's gravity on airborne particles.  This mechanism has
an important influence  on the  deposition of particles  larger  than  0.5 fim D  .
                                                                            36
Settling has an important role in the deposition of environmental aerosols  in the
distal region  of  the bronchial  airways and in the  alveolar region.   Impaction
dominates deposition of particles larger than 3 H*n D  in  the nasopharyngeal and
                                                    3.©
tracheobronchial regions (Pattle,  1961; Bohning et al.,  1975).   In this process,
changes in airstream direction or magnitude of air velocity  streamlines or eddy
components are not followed  by airborne particles  because  of their inertia.  The
passages of the nose  contain smaller airways, and the convective mixing spaces of
the nasal turbinates would be expected to collect  some particles aa small as 1 or
2 urn D    by impaction.   Hence,  impaction is an important process affecting the
      3.6
inhalation deposition  in  the human airways of environmental aerosol particles
>1 jim in aerodynamic diameter.  Deposition by diffusion results from the random
(Brownian) motion  of very  small  particles  caused  by  bombardment  of the  gas
molecules in air.
                                      5-3

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     Particles  larger  than 0.5  |im have relatively  small  diffusional mobility



compared with sedimentation or inertia;  diffusion  primarily affects deposition



of particles with physical diameters smaller than 0.5 ^m.  Interception consists



of noninertial incidental meeting of a particle and the lining of the airway and



thus depends on the physical size of the particle.



     Interception along with electrostatic attraction of particles to the walls



of the respiratory airways are probably minor mechanisms of deposition in most



circumstances.  It is  important  to note that the  diffusivity and interception



potential of a particle depend on its physical size, while the inertial proper-



ties of  settling  and  impaction depend  on  its aerodynamic  diameter.   Chromium



absorption following inhalation  exposures  has received  only limited attention.



The effects of valence state and chemical form on inhalation absorption are not



well defined.   The  problem is further  complicated  by the  inability to clearly



differentiate between pulmonary absorption  and subsequent absorption through the



gastrointestinal tract, and gastrointestinal absorption resulting from clearance



from the respiratory tract followed  by  swallowing.   In  addition,  absorption is



often estimated by urinary excretion, and in many studies fecal contamination of



urine specimens has not been ruled out.



     Langa'rd et al. (1978) exposed  rats  to zinc  chromate(VI)  dust at a concen-



tration of 7.35 mg/m .   In initial  studies whole blood  chromium concentrations



were evaluated following 100,  250, and 350 minutes  of exposure.  Before initia-



tion  of  exposure,  the  mean  blood chromium concentration  was  0.007 ng/mJl.



Following exposure,  values were 0.024, 0.22,  and 0.31  jig/mi  at  100, 250, and 350



minutes,  respectively.   These  data indicate a very rapid pulmonary absorption of



a portion of the inhaled dose.



     The  next portion  of the  study involved four consecutive daily exposures of



6 hours duration to zinc chromate at a level of 7.35 mg/m  .  Blood concentrations
                                      5-U

-------
appeared to peak after the second exposure session.  Mean blood chromium values




from  samples  taken immediately after each exposure  session were:   0.30, 0.56,




0.46,  and  0.34 ng/mS, for  days 1  through 4.   The  mean blood  chromium level




following 2 months of exposure for 6.5 hours/day, 5 days/week was quite similar




(0.495 |ig/m&).    There  were  no   significant  differences   in  absorption,  as




reflected by  blood chromium level, between  the sexes  or between day and night




exposure regimens.




      Baetjer et al. (1959a) exposed guinea pigs via intratracheal administration




to 200 (xg chromium as sodium chromate(VI), potassium dichromate(VI), or chromic




chloride(III), all of which are water soluble salts.  For the Cr(VI)  compounds 10




minutes post-instillation, 15? of the dose remained in the  lungs, 2Q% was found




in the blood,  and  5%  was distributed among various soft tissues.   Clearance of




chromium upward from the trachea and  subsequent swallowing is assumed to account




for the majority of the  dose not found in the  blood and tissues.  During the first




24 hours, = 13? of the dose was excreted,  11? remained in the lungs, and 16$ was




found in the blood and other  tissues.  These  data indicate that as much as 60? of




the administered dose may have  been subsequently ingested.   Similar  data are not




available for  inhalation exposures.   After  140 days,  chromium had been essen-




tially  cleared  from  all  tissues  except  lung  and  spleen.     For  chromic




chloride(III)  10  minutes post-instillation,  69? of the administered  dose was




retained in the lungs, with 4?  in the blood and  other tissues combined.   Twenty-




four hours post-exposure, the lungs retained 45? of the administered dose; lung




retention after 30 and 60 days was 30 and 12?,  respectively.




     Of the water  soluble  salts studied, Cr(III) is absorbed  much more slowly




from  the lungs than Cr(VI)  possibly  as a result  of binding  to  extracellular




macromolecules.  In addition,  analyses  of lungs from  experimental  animals and




human autopsy specimens indicate that water soluble salts undergo conversion to
                                      5-5

-------
very insoluble moieties with long residence times  in lung tissue (Baetjer et al.,



1959a).  The characteristics and potential biological effects of these insoluble



Complexes which are only slowly released from lung tissue are unknown.



     Wada et al.  (1983) exposed male  SD strain  rats  to CrCl  at an atmospheric



concentration of 14.1 mg/m  (Cr),  and observed that the chromium was associated



with both high and low molecular weight proteins.  The chromium which remained in



the lungs was associated with the high molecular weight fraction and this frac-



tion  slowly decreased with  time following  exposure.    The level  of chromium



associated with the  low molecular weight fraction remained constant  for the 5



observation  days  following treatment;  however,  chromium associated  with this



fraction accumulated with  time in the  liver.  The authors suggested that the low



molecular weight protein was involved in the absorption  and transport of chromium



following inhalation.



     Visek  et al.  (1953)   also studied  the  fate of 5 CrCl_ following  intra-



tracheal  instillation.    Seven days  post-exposure,  55%  of  the chromium  was



excreted  in the  feces  and 1%  in  the  urine.    These  data  agree with Baetjer



et al. (1959a),  in that  a  large portion  of the administered dose appears to have



been cleared to the gastrointestinal  tract.  Tissue concentrations in this study



indicated that =5$ of the administered dose was absorbed from the lungs.



     No  data are available which  could be  used to accurately  estimate total



pulmonary absorption following inhalation exposure.   The contribution of gastro-



intestinal absorption to body  burden  following inhalation or intratracheal expo-



sure is not  clear.  The assumption has  been made that gastrointestinal absorption



is so poor that  its  contribution would be negligible.  However, with one estimate



of  pulmonary absorption  at   only  5%  (Visek et al.,  1953),  gastrointestinal



absorption  could  play a significant  role (Section 5.1.2).   Although available
                                      5-6

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data do indicate a significant  effect  of valence state on  absorption,  too few



examples have been studied to allow a thorough understanding of the process.








5.1.2.  Gastrointestinal Absorption of Chromium.  Gastrointestinal absorption of



chromium also appears to be dependent on valence state.  Donaldson and Barreras




(1966) have examined absorption of 51CrCl3(III) and Na251CrOl|(VI) in rats and in



human  patients.   In  conjunction with this  study,  they conducted a  series of



in vitro evaluations to clarify factors affecting absorption.



     Based  on fecal  excretion,  mean  absorption of orally  administered    Cr




compounds  in human  patients  appeared to  be  -Q.H%  for CrCl- and =10.6? for



NapCrOj,.  Approximately  0.5 and 2.1$ of  the doses of CrCl^, respectively, were



excreted  in the  urine  after  2H hours.   When  CrCl-  was  administered intra-



duodenally,  absorption  of  CrCl   was not  appreciably increased; however, intra-



duodenal administration of NaCrOj. resulted in an estimated 50% absorption.



     In rats, *2% of the intragastric dose of both CrCl  and Na-CrOj. appeared to



be absorbed based on  fecal excretion  (i.e., it was assumed that  any radioactivity



not recovered in the feces represented the absorbed portion of the dose, although



it should  be noted that the percision  of these estimates  may be questioned).



Jejunal administration increased  apparent CrCl_  absorption to  8$, while NapCrOj.



was increased to  =25$.   Similar low levels of absorption of  less  than 1$ were



estimated by determining whole body counts 2 days after  administration of radio-
labeled CrCl_ or NagCrOjj (Sayoto et al., 1980).




     IH vitro studies  by Donaldson  and Barreras (1966) showed that CrCl., (Ill)




was  bound  by  both neutralized and  acid  gastric juice, while  Ha-CrO^  (VI)  was



bound  by  acid gastric  juice alone.   Binding  effectively  prevented  uptake by




intestinal rings.  Acid gastric  juice,  in addition to binding Na^rO^ (VI), also




was capable of reducing Cr(VI) compound to Cr(III).
                                      5-7

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     These  absorption  values must  be  considered rough  estimates.   Short-term



urinary excretion values cannot take into account either deposition of chromium



into body sinks or the potential contribution of intestinal excretion.  Absorp-



tion estimates by difference from fecal excretion also cannot take into account



the potential role of enterohepatic cycling.
     Mackenzie et al. (1958) administered either KgCrO^ (VI) or  CrCl, (Ill) in



the  drinking water of  rats at  a  concentration of  25  mg/A.   After  1  year of



exposure, animals  receiving the hexavalent compound had  average  tissue levels



that  were  9 times higher  than those receiving  the  trivalent salt.   This was



interpreted  to indicate much greater gastrointestinal absorption of the hexava-



lent  salt.   The possible  role of differences in  elimination  kinetics was not



addressed.



     Mertz  et  al.  (1965)  administered  a  single dose  of 0.15,   1.0,  or  10 \ig



  CrCl~/g body weight by stomach tube to  rats.   By comparing whole body radio-



activity 4 to  10 days after an intragastric dose with that following an intra-



venous  dose, they suggest 2  to 3% as  a  rough estimate  of  gastrointestinal



absorption.  Absorption was found  to  be independent of  dose  level  or dietary



chromium history (deficient versus supplemented).



     Visek et  al.  (1953)  gave  a single dose  of  CrCl  (III) by gavage to  rats.



Tissue distribution indicated <0,5% absorption after 4 days.  Urinary excretion



suggested higher  absorption estimates,  but fecal  contamination  of the samples



was suspected.



     MacKenzie  et  al.  (1959)  administered Na?CrOj,  (VI)  as a single  dose by



stomach tube.  Urinary excretion of 6$ of the dose after  14 days in fasted rats



and 3$  in  non- fasted  rats was observed and used as  an  estimate of absorption.
Blood chromium concentrations when  rats were given Na^rO^  were 2-fold higher


                       51
than for animals  given  CrCl_.  This is not surprising,  since Cr(III) is cleared
                                      5-8

-------
 from the  serum much more rapidly than Cr(VI) disappears from the erythrocytes.

 Urinary excretion data  for Cr(III) compound were not reported.

     Additional evidence for the poor absorption of CrCl_  (III) was presented by

 Mertz  et  al.  (1965).  They found  that a  100-fold greater oral dose was required

 to  alter  glucose tolerance in rats than  the intravenous effective dose.

     Ogawa  (1976) examined differences in absorption and metabolism via several

 routes of exposure  using CrCl (III) and  Na?CrO  (VI) administered to rats.  They

 found  gastrointestinal  absorption to be  2.4 and 1.4$, respectively, for the two

 salts.   When  animals were  fasted for 48 hours, absorption of both  salts was

 increased to  11$.

     Although  precise quantitation of  absorption  efficiency is  impossible,  a

 reasonable  estimate from the  available  literature  discussed above is that both

 valence states are  absorbed at efficiences of <5%.




 5.1.3.    Chromium  Absorption Through  the  Skin.   Percutaneous  absorption  of

 chromium  appears to be  related  to valence  state, the particular salt employed,

 and the concentration applied.

     Mali (1963) conducted a series of in vitro  and in vivo  studies to determine

 penetration of potassium dichromate(VI) and  chromic sulfate(III)  through the

 skin (the source of skin was  not mentioned).   They found that neither compound

 diffused  spontaneously  through  intact,  isolated   epidermal membranes.    The

 diffusion  constant  for  diffusion  of   the  Cr(VI)  compounds  through  dermis
          C    *)
 (314 x 10~  cm /min)  indicated  unimpeded  absorption;  however,  the  diffusion
                                                  C   2
constant  for the trivalent compound was  26.6 x 10   cm /min.

     Significant amounts (30$) of Cr(III) were bound to dermal proteins in vitro,

while  only  very  small amounts (1$) of  Cr(VI) were bound (Mali,  1963).   These

results were confirmed in human volunteers.  It was found that in vivo potassium
                                      5-9

-------
dichromate but not  chromic  sulfate penetrated intact epidermis.   In addition,


reduction of Cr(VI) to Cr(III) in the skin tissue was demonstrated.  The rate of


this reduction was pH dependent.


     Samitz and Katz (1963,  196*0 presented additional data which indicate that


Cr(III) binds to skin in  vitro,  and that binding following  exposure  to Cr(VI)


salts is dependent upon reduction to Cr(III).


     Samitz and  Gross  (1961)  presented  preliminary  evidence that there  is  no


difference  in  absorption of  potassium  dichromate(VI)  as compared  to chromium


nitrate(III) in vivo in guinea pigs.  Samitz  and Shrager (1966)  reported addi-


tional evidence that permeability of the skin to Cr(III)  is dependent upon which


salt is employed.   Their data indicate  that  absorption of  chromic  sulfate  is


negligible, absorption  of  chromium nitrate  is  somewhat greater  than  chromic


sulfate absorption, and absorption of chromic chloride is as great as potassium


dichromate.


     Wahlberg (1965) reported that absorption of a given salt is dependent upon


the concentration applied.   In this  study,  application of  0.017 M  to  0.239 M


solutions of Cr(III) resulted  in percentages of absorption which were not statis-


tically different  from absorption  experiments  under the same  conditions  for


Cr(VI).  However, at concentrations from 0.261 to 0.398 M, significantly more of


the Cr(VI) compound  was absorbed.  Absolute absorption rates for chromic chloride


were maximal with application of 0.239 to 0.261 M solutions, reaching levels of

                             2
315 to 330 n moles Cr/hour/cm  .   In contrast, absorption  rates of sodium dichro-

                                                        2
mate reached a maximum of 690 to ?25n moles   Cr/hour/cm  at a concentration of


0.261 to 0.398 M.


     In conclusion,  percutaneous absorption of chromium  is dependent not only on


valence state,  but is also affected by the anion, as well as the concentration


and pH of the  applied solution.
                                     5-10

-------
5.2.  CHROMIUM TRANSPORT,  METABOLISM,  DISTRIBUTION,  AND ELIMINATION








5.2.1.  Transport and Metabolism.  The mechanism of chromium transport is depen-




dent upon  the  valence state  after  reaching the  bloodstream.   Cr(VI)  readily




crosses erythrocyte membranes. Kitagawa et  al. (1982) demonstrated in vitro the




inhibition of  Cr(VI)  penetration  into erythrocytes  by  U-acetamido-4'-isothio-




cyano-stilbene-2,2'-disulfonic acid,  an  inhibitor  of  anion transport.   After




entry, the Cr(VI)  undergoes  reduction to Cr(III)  and  this reduction has  been




investigated in  other cell systems.   In  studies with  cultured hamster fibro-




blasts, Levis  et al.   (1978)  have shown  that only Cr(III)  could  be  identified




intracellularly following  treatment  with potassium  dichromate.   The mechanism




for intracellular reduction is not completely understood.  Several  investigators




have proposed a role for the  mixed function  oxidases.  These proposals are based




upon  the  reduction  of mutagenic  activity  following   addition  of  microsomal




preparations to Cr(VI)  (Gruber and Jennette,  1978;  Lofroth,  1978;  Petrilli and




DeFlora, 1978a).   While rat  liver  preparations are  very  effective in reducing




mutagenic activity, lung preparations show  only  minimal activity  (Petrilli and




DeFlora, 1978b).   Addition of reduced  cofactors, such  as GSH, NADH,  or NADPH,




also  resulted  in  reduced  activity.   LSngard (1979)  presented  evidence  that




although reduction of Cr(VI) may take  place at  any intracellular  site  where




electron donors are available (which includes the microsomes),  the primary site




of  reduction is  within the mitochondria.    In  relation to  chromium  binding to




hemoglobin, Kitagawa  et al.   (1982) reported  that  the  reduced  cofactor GSH was




necessary for the in vitro binding of Cr(VI) to purified hemoglobin,  suggesting




that either the chromium or the hemoglobin has been reduced for binding to occur.




Evidence that the reduction of Cr(VI) to Cr(III) may be  of biological  importance




was provided by Aaseth et  al. (1982).   Cr(VI) was shown to bind to erythrocytes
                                     5-11

-------
under in  vitro  incubation conditions.   When GSH was  added to the  incubation



media, the binding of chromium to the erythrocytes decreased as a result of the



reduction of Cr(VI) to Cr(III) and the inability of Cr(III) to penetrate the cell




membrane.  Reduction  of chromium binding was also  observed when intracellular GSH



was decreased by the action  of diethylmaleate.   In this case, it  was  proposed



that the lower binding resulted from the failure  to reduce Cr(VI)  to Cr(IIl) in



the cytosol.  Jennette (1982) demonstrated that  a Cr(V) intermediate was formed



during the iri vitro  reduction of  Cr(VI)  to Cr(III)  by rat liver  microsomes and



that this intermediate may be the chemically reactive form of chromium.



     Localization In vivo of Cr(III)  within the body cells of the rat appears to



be  time  dependent,  with  initial high  concentrations within  the cytosol  and



subsequent translocation to mitochondrial and nuclear fractions (LSngard, 1979).



In  addition,  the partitioning  of intracellular  chromium appears  to  be  dose



related.  Tandon et al.  (1979)  found  that chromium nitrate(III) doses of 1, 2 or



3 mg/kg administered intraperitoneally  resulted  in  an increased  percentage in



the mitochondrial  fraction  with increasing  dose.    Early hypotheses  assumed



intracellular binding of Cr(III) to proteins; however,  in vitro data developed by



Sanderson (1976) indicated that intracellular chromic ion exists  in the form of



coordination complexes with small organic anions.



     Transport of Cr(III)  is facilitated by specific  binding with siderophilin



(Hopkins and Schwartz, 196U).  Following very large doses,  binding  with  other



serum proteins occurs.  Red blood cells appear to be essentially  impermeable to



Cr(III).   Impermeability  of  all  cell membranes  to  Cr(III) compounds has  been



traditionally accepted;  however,  Levis et al. (1978)  have shown uptake of CrCl_



by cultured hamster fibroblasts.   In  addition, Tandon  et  al.  (1979)  have  shown



significant  intracellular levels of chromium  following in  vivo exposure  to



Cr(III) nitrate.
                                     5-12

-------
     A  number  of  investigators  have  demonstrated  that  in  pregnant  rodents




exposed to inorganic  chromium only a very small fraction  of the administered dose




is transported to the fetus in  utero.  Visek et al. (1953) gave   Cr,  as a single




dose, in the form of  sodium chromate(VI)  or chromium chloride(III) intravenously




to rats on days  15 to 20 of gestation.  Fifty  (iCi per dose was administered, and




specific activity ranged from 200 to 1250 |iCi/mg.   Litters were examined 2*1 hours




post-injection.  Regardless of chemical  form or time of  administration, recovery




of ^1Cr per total litter never exceeded  0.13$ of the administered dose.




     Mertz et al.  (1969)  administered  5 Cr  as chromium acetate(III)  to rats.




Single doses were given  either intravenously  or  by gavage  at mating.  Repeated




doses were given either by  daily intubation during gestation or by administration




in the drinking water, at a concentration of 2 mg/8,.  Five (iCi were given intra-




venously and either 5 or 250 |iCi by gavage.  Specific activity ranged from 30 to




100 (iCi/iig.  Pups were sacrificed no later than 3 hours following birth.




     No   Cr was detected in the young at birth following a  single dose by either




route to the  dam at mating. Repeated  dosing by gavage during gestation resulted




in labeling of the litters  at birth with from 0.5 to 1.5% of the mothers' total




body activity.   Exposure  of  dams  via  the drinking  water  did  not result  in



            51
transfer of   Cr to the litters.




     Matsumoto et al. (1976) examined placental transfer following subcutaneous




administration of CrCl-(III) to ICR mice.  Six control  mice were injected with




saline, 11 mice  were injected  with CrCl- (as  Cr)  at a  dose of 9-76 mg/kg,  and




6 mice were injected with  19.52 mg/kg.   Mice were injected  9 times every other




day from the  first to the sixteenth day of gestation.




     Although there appeared to be  a  trend  of increasing chromium  in  the pups




with dose, levels were not  significantly different  from  controls for either dose
                                     5-13

-------
group.  The high  dose  dams  did exhibit placental chromium  concentrations that



were significantly elevated above controls.


                                       51
     In contrast to these studies, when   Cr was administered to rats in the form



of glucose tolerance factor (a low molecular weight organic complex isolated from



yeast) by stomach tube  during gestation,  from 20 to  50%  of  the  dam's  radio-



activity was detected in the litters  (Mertz et al.,  1969).   This is consistent



with the role of chromium as an essential trace element.



     In  conclusion,  there are  limited data  that indicate  transfer of  small



amounts of chromium from mother to offspring when chromium is administered as a



salt  during  gestation.   This appears  to  be true for  both Cr(III) and  Cr(VI)



salts.   These  data lend support to a  hypothesis  for chromium teratology which



relies on  a  mechanism  other than direct  interaction between  chromium and the



fetus.







5.2.2.    Distribution.   Once  absorbed,  Cr(III) compounds  are cleared  rapidly



from the blood  and more slowly from the tissues.  This also indicates that blood-



chromium  levels are inadequate indicators  of  body burden.   Clearance of Cr(VI)



from the  blood is  slower, presumably  due  to uptake by erythrocytes followed by



reduction to the relatively impermeable Cr(III).



     Hopkins (1965)  injected  0.1 pig   Cr  (as chromium  chloride )/100 g  intra-



venously. The blood chromium content as a  percent of  the  15 minute blood concen-



tration  at  various time intervals was: 30 minutes,   94$;  1  hour, 87%; 2 hours,



69%;  4 hours,  66%; 8 hours,  47*;  24 hours,  17*; 48 hours,  9%;  96 hours, 5%.



Withey  (1983)  replotted this  on semi-log paper  and  resolved the  curve into



initial,  intermediate, and terminal phases with respective  half-lives of  0.56,



5.53 and 57 hours (Figure 5-1).
                                      5-14

-------
 I
8
   <

   Q.
   o
   z
   iu
   CC
   »-
   Z
   uu
   o
   CC
   UJ
   CL
                       t %  = 5.53 HOURS
                  =  57 HOURS
           t %  = 0 .56    \
                 HOURS  \
                  I
                  20
 I
40
 I
60
 I
80
 1
100
                     HOURS AFTER INJECTION
   Figure  5-1.  Rate of blood clearance of intravenously injected   Cr (III)
               from male rats.  Data from Hopkins (1965) as replotted by
               Whitney (1983).
                              5-15

-------
     Visek  et  al.  (1953)  compared clearance and  distribution  following intra-



venous injection of several chromium salts in rats.  Four days following injec-



tion, NaCrO^ (III) blood levels were <0.02$  of  the  administered dose per gram of



blood and  CrCl-  (III)  levels were 0.05$  of  the  administered dose per  gram  of




blood.  In  contrast, Na^rO^VI) levels 4 days  post-injection represented 0.5255



of the administered dose per  gram  of blood.  Nearly complete blood clearance for



this salt was not achieved until H2 days post-injection compared with only 7 days



for the trivalent salt.




     Baetjer et  al.  (1959a)  administered Na^rO^  (VI), K2Cr207  (VI),  or CrCl3



(III) intravenously to  guinea pigs.  For the Cr(VI) salts, values of \ig Cr/10 gm



dry tissue/200 |ig Cr injected for erythrocytes  1  and 3 days post-injection were



28 and 31,  respectively, and  for plasma, 4 and 2,  respectively.   For the Cr(III)



salt, erythrocyte values at  the  1 and  3  day time points were 1  and <1,  respec-



tively, and plasma values were 5 and 3» respectively.



     Visek  et  al.  (1953)  have reported organ distribution of several  chromium



salts following intravenous injection in rats. Sodium chromite (NaCrCO (VI) was



concentrated in  large  quantities by  the reticuloendothelial system, which  in



combination with  the liver  accumulated  90$ of  the dose.   At  42 days  post-



exposure,  organs with  detectable  levels  were:   spleen  > liver  > bone  marrow >



tibia epiphysis > lung  > kidney.  The liver and spleen  contained 33  and 50$ of



their U day values, while the lung contained  10$.   The extensive accumulation of



chromite in the reticuloendothelial system is postulated to be the result of the



formation of colloids by chromite at physiological pH.



     Chromic chloride concentrated in  the liver,  spleen and bone  marrow;  once



deposited,   it  cleared  very  slowly.    At 4  days  post-exposure, CrCl   exposed



animals had lower percentages of the  total  dose  in their livers,  spleens,  and



bone marrow than  those  exposed to  NaCrOp.  More CrCl- than NaCrO? accumulated in
                                     5-16

-------
the kidney, however.  All organs gradually  cleared  chromite  over the period of



the study.  In  contrast, the  liver  in CrCl--exposed rats was the only organ to



clear significant amounts of chromium over the study period (45 days).



     Cr(VI) was  picked  up  to a much  smaller  extent  than either of the Cr(III)



salts.  At the 42-day time point, <1? of the  dose was  found  in  the liver.   For



comparison with the  trivalent  salts, 4-day  tissue concentrations  in animals



given sodium chromate were  (% dose/g tissue):  liver, 0.52%; spleen, 0.91?;  bone



marrow, 0.56?;  tibia  epiphysis,  0.38$.   At  42 days,  corresponding values were:



liver, 0.07?; spleen, 4.8?;  bone marrow, 0.16?; tibia epiphysis,  <0.02?.   The



increase in spleen concentration was attributed to erythrocyte destruction.



     Hopkins (1965) examined the kinetics of distribution with trace quantities



of  chromic chloride in the  rat.   Measured concentrations at  15 minutes post-



injection were highest in lung and  kidney,  followed  by heart,  pancreas,  liver,



bone marrow, spleen,  testis, and brain.   The  heart,  lung, pancreas, liver, and



brain  showed  maximal levels  at this  15-minute time  point, and  subsequently



declined.  The  spleen reached a maximum level 96 hours  post-exposure and the



testis 4 hours  post-exposure  (300  times the  initial level), while  the  kidney



concentration remained unchanged over 96 hours.



     Following intratracheal administration  of chromate to rats, highest concen-



trations were found in the lung,  followed by liver,  kidney, and spleen (Baetjer



et al., 1959a).   In contrast to the injection  studies,  significant concentra-



tions were not found in bone.



     Following a single oral  dose  of chromate administered  by  gavage, highest



concentrations were found in the liver,  followed  by the kidney  and  the  spleen



(MacKenzie et  al.,  1959).   When chromate was administered in the drinking water



for 1 year, highest  concentrations  were found in the  spleen,  followed  by the
                                     5-17

-------
kidney, liver, and bone.  Year long administration of chromic chloride resulted




in the same type of distribution pattern with lower absolute quantities.



     Generalizations regarding behavior of the two valence states are difficult



to make, since almost all testing has involved chromic chloride(III) and sodium



chromate(VI) or  potassium dichromate(VI).    The  data of  Visek et al.   (1953)



showed  differences  in  uptake and  distribution  between  chromic chloride  and



chromite; the behavior of other Cr(III) salts is difficult to anticipate.  Data



from chromium chloride(III)  do indicate that  once  absorbed, this salt is quickly



cleared from the bloodstream and has a long tissue residence time.  Cr(VI) salts



have longer residence times in the  blood  and are  more rapidly cleared from the



tissues.  Comparisons of  tissue distributions  are difficult,  because different



authors choose different organs to examine and time between exposure and analysis



varied considerably.  In almost all instances, the following organs accumulated



significant amounts  of  chromium:  liver, kidney,  spleen,  and  bone  marrow.   In



addition, there are some data which indicate  that the testis,  heart, lung,  and



brain accumulate considerable  amounts.  Tissue distribution does not  appear to be




affected by the  sex of  the animal  or  by  dietary  history in terms  of chromium



content.



     In humans,  Teraoka (1981) studied the  distribution  of 24  elements in 12



Japanese males.  The samples obtained at autopsy were from seven workers exposed



to  heavy  metals and five subjects  with no undue  exposure.   Although absolute



concentrations of chromium varied extensively between individuals, the greatest



concentrations of chromium were found in the Hilar lymph nodes and lungs followed



by spleen, liver, kidney, and heart which  all contained approximately the same



level for each individual.  The distribution  patterns  were similar for the seven



men occupationally  exposed  to metals  and the unexposed control  group.   The



average level of chromium in the Hilar lymph nodes of unexposed men was 8.2 ppm
                                     5-18

-------
 as  compared  with  2400 ppm for the two chromium plating workers and  152 ppm for



 the three chromate refinery workers.  An airplane painter and stone mason who had



 possible exposure to  chromium had levels of 33 and 4.2 ppm, respectively.  The



 order  of organ distribution  appeared to be  similar  regardless of whether the



 exposure to  chromium  was from occupational (high levels) or environmental (low



 levels) sources.



     Similarly, the highest  levels of chromium were detected at autopsy in the



 lungs  of a worker who  died of respiratory cancer 10 years after retiring from a



 chromate  manufacturing  plant  (Hyodo et al.,  1980).    The  chromium  levels  in



 different portions of  the lungs  of this worker varied from  616 to 7100 ng Cr/g



 wet weight  as compared  to values  of 19.3  to 881 ng Cr/g wet weight of similar



 lung sections from five control subjects. The suprarenal gland, brain, and skin



 also had relatively high levels of chromium as compared to the control values.



 It  was reported that Cr(VI) was more  prevalent in the chromate worker; however,



 the wet  oxidation method, using hot nitric acid and  perchloric  acid,  has the



 potential of oxidizing Cr(III) to Cr(VI) and makes these results invalid.








 5.2.3.  Elimination.  Hopkins (1965)  examined  the kinetics of single doses of 0.1



 or  0.01  \ig  Cr administered intravenously  as   CrCr_.   Urinary  excretion  was



 followed  over 4  days,  and  an inspection  of a  semi-log  plot  of  these  data



 indicated at least two  elimination  components, the  first  representing  rapid



 blood  clearance  of chromium  and the second  the  slower elimination  from  soft



 tissues.  Hopkins data also indicate that,  although  the urine is  the  primary



 route of excretion, the intestines also play a small role.




     Mertz et al.  (1965) followed  the  elimination of  single  doses  of intra-



venously administered  chromic chloride over  a longer time period  (72 days).



Whole body retention  of  chromium  as a  funtion  of time was represented by  a
                                     5-19

-------
polynomial with three  distinct regression terms  (Figure 5-2).  This  evidence



supports a three compartment elimination process.  The half-lives for chromium in



the  three  compartments were  estimated from  clearance  rates  to be  0.5 days,



5.9 days, and 83.4 days.  At the end of 72 days, 13.5%  of the original dose was




retained.



     Onkelinx (1977) also  followed  elimination of  an  intravenous dose of chromic



chloride.   Their  data agree  with Mertz  et al.   (1965),  in  that  the  plasma



disappearance curve  could  be  described by the  sum of  three exponential compo-




nents,  representing  three  compartments.  Excretion  has been  quantified  and 3



components have been identified. Urinary  excretion represented 51 to 64$ of the



total excretion, fecal excretion 5  to 8%,  and clearance of chromium into a body



"sink"  31  to  41$.   The sink represents compartments with extremely long half-




lives .



     In a  comparison of Cr(VI) and Cr(III) elimination in rats,  Sayoto et al.



(1980)  administered  radiolabeled Na2CrO|j or CrCl- to animals either by intuba-




tion or intravenous  injection.  Regardless of  the route of administration, the



Cr(VI)  compound was excreted more  rapidly through the  feces  and urine.   The




biologic half life for Na CrOj, and CrCl- after intubation was 22.24 and 91.79



days, respectively.   The  authors  suggested that  these  findings  indicated that



Cr(III) has a higher affinity for body constituents than Cr(VI).



     Yamaguchi  et  al.  (1983)  studied the excretion of  a Cr(III)  and Cr(VI)



following subcutaneous administration to rats of Cr(NO )_ and KpCr^O™.  Within 24



hours after treatment 93.6$ of  the  Cr(VI) was removed from the site of injection



while only  21.3$  of the Cr(III) was  adsorbed.  In 7 days,  48 and  8.0$ of the



Cr(VI)  and Cr(III),  respectively,  were eliminated in the  urine.   The biologic



half-time of Cr(VI) in different organs was determined, with half-times for the



terminal  component for lungs,  liver, kidney,  brain,  heart,  testes  and blood



calculated to be 20.9, 15.7, 10.5,  9.6, 13.9, 12.9 and  13.9 days, respectively.





                                      5-20

-------
  100
   20
ui
U
cc
UJ
0.
   10
1 \
K-
j
t % = 5.9 DAYS
V 	 t% = 0.5
1 1
10 20
~^-^_
DAYS
I I
30 40
t % = 83. 4 DAYS
"""»"*»•-. ^^__
!P^— ^
I I I
50 60 70
~-—-
I
80
                              DAYS AFTER INJECTION
    Figure 5-2*1 Whole-body elimination of intravenously administered
              ^ Cr (III) in male rats (Mertz et al., 1965).
                                   5-21

-------
     Collins  et  al.  (1961) examined  chromium excretion  in dogs  dosed  intra-



venously with chromium chloride  or sodium dichromate.  Acute exposures (dose not



stated) showed  that 25%  of  the Cr(III) salt and  9$  of the Cr(VI)  salt were



excreted in the urine within 4 hours.   Less than 0.5% appeared in the bile.



     In bile fistulated dogs,  1-day  excretion  values were  50% in the urine, 0.5%



in the bile,  and  3.7% in the feces for  the Cr(III) salt, and 20, 0.9, and 1.2$ of



the dose in urine, bile, and feces,  respectively, for the  Cr(VI) salt.  Excreted



chromium was readily dialyzable indicating  that,  if the chromium was bound, the



molecule it was  combined  with was of  small size  or the binding was reversible.



Tubular reabsorption appeared  to represent  >63/& of the amount filtered.




     Davidson et al. (197*0 examined kidney handling of chromium in normal human



subjects and in  dogs  (not pre-treated with chromium).  Their  results indicate



that  in both dogs  and man physiological  quantities of  chromium have a >99/6



reabsorption value.  Their data also  suggest  that  there  is  an  active transport



mechanism in the renal tubule  for chromium  reabsorption.   They hypothesize that



the normal filtered load  for chromium may  be  close  to  the maximal reabsorption



rate.   If this is  the  case, then  any  increase in plasma chromium concentration



would result in a significant  increase in renal excretion of chromium.








5.3. SUMMARY








     There are only limited experimental data available on the pharmacokinetics



of  chromium.    Absorption by  inhalation   exposure  appears  to occur  rapidly,



although it  is difficult  to quantify  the  extent of absorption.   A preliminary



estimate of pulmonary absorption,  following disposition of CrCl  into the lungs



by instillation, indicates that approximately 5% is absorbed.   Following oral



exposure,  gastrointestinal absorption was also low with  estimates that only 5% or
                                     5-22

-------
less of chromium was absorbed.  In vitro studies indicate that gastrointestinal




juices have  the capability to  reduce  Cr(VI)  to Cr(III);  however, there  are



insufficient data  from  in vivo studies to demonstrate whether  this  reduction



process has  the capacity  to  eliminate any  differences in absorption  between



ingested Cr(VI)  and Cr(III)  compounds.   Percutaneous absorption  of  chromium



through unbroken  skin  is  variable and dependent on  valence as  well as  the




specific salt.



     After absorption, chromium is transported by the blood with Cr(III) trans-



port facilitated by specific binding proteins in the blood.  Cr(VI) on entering



the blood  stream  diffuses  into the blood  cells  where  reduction  and binding to



cellular components occurs. Both absorbed Cr(III) and (VI) can be transported to



a limited extent to the fetus in utero after  exposures  of the dams, although the



data do not allow quantitative estimates of fetal  exposure.  Chromium transported



by  the blood  is  distributed to  other organs with  greatest  retention by  the



spleen, liver, and bone marrow.  The major deposition site following inhalation



exposure is  the  lungs,  where chromium  probably  binds  to the  cellular material




before absorption can occur.



     Absorbed chromium is eliminated from the body in a rapid phase representing



clearance  from the blood  and in  a slower  phase representing  clearance from



tissues.   Urinary  excretion  is  the primary route of elimination accounting for



somewhat over 50$ of the eliminated chromium, while fecal excretion accounts for



only 5% of the elimination from the blood.  The remaining chromium is deposited



into deep  body compartments,  such as  bone and soft tissue.   Elimination from



these tissues proceeds very slowly.  Estimated half-lives for whole body chromium



elimination are 22 and 92 days for Cr(VI) and Cr(III), respectively.
                                      5-23

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                      6.  CHROMIUM AS AN ESSENTIAL ELEMENT








 6.1. CHROMIUM DEFIENCY








     The nutritional need for chromium as an essential  element has been reviewed




 by  Golden  and Golden (1981), Anderson  (1981),  Saner  (1980) and Mertz (1974).




 Mertz  (197^)  described  the necessary components  for considering an element as




 essential.  These components  are: the element must be found in living matter, the




 element  must interact  with  living biological  systems,  and deficiency  of the




 element must  produce  a  decrement in biological  function.   Indeed,  chromium is




 found  in  living matter, it  does interact with  living systems,  and deficiency




 syndromes are remedied with its supplementation.




     The National Academy of Science (NAS,  1980)  has summarized daily intakes of




 chromium  by persons  in the  United States  as:    dietary,  a  range  of 37  to




 130 ng/day  with an average of 62 jig/day;  air,  a range of <0.5  to  <4.0  of the




 dietary intake;  and drinking water, a range of 0  to 224  ug/day with an average of




 17 jig/day.  Mertz (1974) provided similar estimates that daily intake of chromium




 in healthy humans was between 5 and 100 jig/day and that this intake resulted in




 blood and urine levels  of  chromium  of 0.5 to 5 and 5 to  10  ng/£, respectively.




 Infants receive chromium through  breast  milk during nursing (Kumpulainen et  al.,




 (1980).   These  levels  of chromium  intake must  be  adequate  since no  serious




 effects from chromium deficiency have been observed in the  general populace.   The




Estimated Adequate and Safe Intake  (EASI)  values for chromium,  provided  by the




National Academy of Science (NAS,  1980),  are listed in Table 6-1.  The lower  EASI




values are based on the  average United States daily intake from a  mixed diet.  No




Recommended Daily Allowance (RDA) has been issued.
                                      6-1

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                                TABLE 6-1




          Estimated Adequate  and Safe Intake  (EASI) for Chromium*








                    Age (years)               EASI  (mg/day)
Infants

Children



0
0
1
H
1
>
.0
.5
to
to
to
11
to 0.5
to 1.0
3
6
10

0
0
0
0
0
0
.01
.02
.02
.03
.05
.05
to
to
to
to
to
to
0
0
0
0
0
0
.04
.06
.08
.12
.20
.20
Adults                                      0.05 to 0.20
•Source:   NAS,  1980
                                        6-2

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     Animal studies have been conducted  to  determine the effects of ingestion of




dietary chromium.  Schroeder et al.  (1964) maintained 54 male and 54 female mice




for life  on  drinking water  containing  5 ppm of Cr(III).  The  mice  exposed to




chromium had survival rates  similar  to  those of control animals  and mean body




weights that were 123/& of control values.   In this study the food was devoid of




all trace metals. In an identical study  using groups of 50 male and female Long-




Evans rats, Schroeder et al. (1965) observed increased longevity in the treated




animals.   In  studies of the effects of the  absence  of chromium  in  the diet,




Roginski  and  Mertz  (1969)  raised rats  in  plastic  cages on  low  protein diets




containing less  than 100 ppb chromium.   A second group  of rats were  given a




chromium  supplement of  2  ppm chromium in the  drinking  water.   The chromium




supplemented rats  had  better weight gain than  the  chromium deficient animals.




This difference in weight gain was more  striking if the animals were allowed free




access  to an exercise  wheel.   When extreme  care  was  taken to  also  prevent




airborne  exposure  to chromium of rats  maintained on  chromium deficient diets,




there was a high incidence of moderate hyperglycemia and glycosuria as compared




to  animals  on chromium  supplemented  diets.   These studies  suggest  that small




amounts of dietary chromium were beneficial to the health  of these  rats.  Preston




et al.  (1976)  maintained  female  guinea pigs  on low  protein  diets  containing




0.125, 0.5, or 50 ppm chromium along with  adequate  levels of vitamins and some




other  trace  metals.  After  8 to  13  weeks  of maintenance on  these  diets, the




animals were mated and allowed to deliver pups.  Although  the  diets had no effect




on  the survival  of animals  not mated, there was a significant decrease in




survival  of mated  animals  on the low chromium diet (13 of  24  animals died) as




compared  to animals on the low chromium  supplemented diet  (4  of  24 animals died)




and the high chromium supplemented diet (3  of 24 animals died).   It was suggested




that  the  added  stress of  mating and pregnancy  along with  chromium deficiency
                                      6-3

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resulted in  the  high level of mortality.   Other parameters  examined included




weight gain  and  glucose loading;  serum cholesterol levels were  not altered by




dietary chromium.  It was concluded that chromium was beneficial  to survival in



guinea pigs as was previously reported for mice and rats.








6.2. GLUCOSE TOLERANCE FACTOR








     Chromium deficiency in the diet results in glucose intolerance in both the




experimental animals and humans  (Saner,  1980).   Schwarz and Mertz  (1959)  have




identified a glucose tolerance factor (GTF) which was required in rats to main-




tain normal rates of glucose removal from the blood stream.  This factor has been




obtained  from  natural  sources including  brewer's yeast  and hydrolyzed  pork




kidney.  In  further  studies,  Mertz and  Schwarz  (1959) demonstrated  that  rats




maintained on  some commercial diets  had normal  glucose removal rates,  while




maintenance on other diets resulted in poor  removal of  glucose from the blood.




When the second diets were  supplemented with GTF, glucose removal  rates returned




to normal values. Mertz and Schwarz (1959) have identified Cr(III) as the active




agent in the GTF.  It was further demonstrated that administration of a variety



of Cr(III) compounds to glucose intolerant  rats resulted in  an increase in the




rate of  removal  of  glucose  from  the  blood.  In  this assay, the  less stable




Cr(III) compounds were  most  effective,  and some Cr(VI)  compounds were totally



without activity.




     The effects of chromium on glucose metabolism were suggested to result from




chromium being a co-factor for insulin.   Mertz et al.  (1965) measured insulin




stimulated  CO    production  in  adipose  tissue  from   chromium  deficient  rats




receiving supplements of 0.0,  0.01, 0.05, or 0.1 [ig Cr/100 g  body weight.   The




adipose tissue of  rats  receiving 0.05 ng Cr/100 g produced  significantly  more
                                      6-4

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COp  than  tissues  obtained from other groups.   Farkas  and Roberson (1965)  per-




formed  similar studies with measurements of  glucose  utilization in rat lenses




taken from  animals  on  chromium deficient and supplemented diets.  The chromium




treatment alone did not affect glucose utilization; however, chromium treatment




in conjunction with insulin significantly increased the utilization of glucose.




Administration of 2 ppm  of chromium in the drinking water of rats also facili-




tated the insulin transport of amino acids into the heart and the incorporation




of amino  acids into protein.   Providing chromium supplements to rats increased




the  animals' sensitivity  to many of the  effects of insulin.




     Saner  (1980) states  that certain groups of persons may be prone to chromium




deficiency;  these  groups  include  the   elderly,  diabetics,  pregnant  women,




malnourished children, offspring and  siblings of diabetics,  persons with early



coronary  heart disease and  their  offspring.   Jeejeebhoy et  al.  (1977)  have




described a female  patient placed on total parenteral nutrition for 3 1/2 years




in whom chromium deficiency was indicated.  Blood chromium levels were reported




as 0.55  ng/m£,  and  hair  chromium  levels as  154 to 175 ng/gm.   In addition to




glucose  intolerance,   weight  loss,  neuropathy,  elevated fatty  acid  levels,




reduced respiratory quotient,  and abnormal  nitrogen metabolism  were reported.




Daily administration of 250 (ig chromium  chloride  for 2 weeks in  the parenteral




infusate, followed by  20  jig/day maintenance,  resulted in a reversal of symptoms.




Freund et al.  (1979) report similar finding of chromium deficiency in a patient




receiving total parenteral nutrition.  Supplementation of the infusate with 150




|ig chromium/day resulted  in  the reversal of  the adverse clinical  findings  of




impaired glucose tolerance, weight  loss,  and confusion.




     Chromium has  been  known to affect glucose tolerance  in humans.  Levin et al.




(1968)  performed  oral  glucose  tolerance tests  on 9  male and 6  female  elderly




subjects.   Of the  subjects tested,  10 daily dietary supplements with 150  fig/day
                                      6-5

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 chromium for 3 to 4 months resulted in normal glucose tolerance in four subjects.




 The remaining six subjects still had abnormal tests after receiving the  chromium




 supplement.  Glinsmann and  Mertz (1966) observed improved glucose  tolerance in




 three  of six diabetics  given  150 to  1000 |ig of Cr(III) for 15 to 120 days.  In a




 similar study, Sherman et al. (1968) observed no  change in the  glucose tolerance




 of ten diabetics who received chromium supplements of 100 |ig/day  for 16 weeks.




 In malnourished infants, Gurson  and Saner  (1971) and  Hopkins et  al. (1968)




 observed dramatic increases in glucose  removal and utilization  following treat-




 ment with Cr(III).  To ensure that  the improved glucose tolerance was the result




 of chromium therapy,  Hopkins  et al.  (1968)  treated five infants  in  a manner




 similar to  the  others with the  exception  that  chromium was  not administered.




 These  five infants showed no improvement in glucose tolerance.   Dieatary chromium




 appears to have some effect  on human glucose tolerance; however, the therapeutic




 effect of chromium  supplementation on subjects  with abnormal glucose tolerance



 was  variable.








 6.3. SUMMARY








     Animal studies have demonstrated that chromium deficient rodents gain less




 weight and have a shorter lifespan than animals maintained on a diet containing




 adequate chromium levels. Chromium deficiency results in glucose intolerance in




 rats and this intolerance can be  reversed  by dietary treatment  with  Cr(Ill).




More effective in reversing glucose intolerance is a chromium complex which has




 been isolated from brewer's yeast  and  designated as glucose  tolerance factor




 (GTF).  GTF may be formed in mammals following ingestion  of  inorganic chromium.




Some humans  with abnormal glucose tolerance, such as the elderly, diabetics, and




malnourished  infants  have   responded  to   dietary  supplements  of  chromium.
                                      6-6

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Although the exact level of chromium needed for good health is not known,  it is




assumed from the lack of observed effects of chromium deficiency that the average




American intake of 0.05 to 0.2 mg/day is  adequate.
                                    6-7

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                            7.  CHROMIUM TOXICOLOGY








7.1. ACUTE EFFECTS OF CHROMIUM EXPOSURE IN MAN AND ANIMALS








7.1.1.  Hunan Studies.  Chromium metal is biologically inert and does  not produce



toxic or  other harmful  effects in man  or laboratory  animals.   When in contact



with the skin, compounds of chromium in the trivalent state combine with proteins



in the superficial layers, but do not cause ulceration  (NAS, 1974).



     Cr(VI) compounds are responsible  for the majority of the health problems



associated with all chromium compounds.   They are irritating and  corrosive, and



may be  absorbed  cutaneously, by ingestion,  or  by inhalation.   Acute systemic



poisoning is rare; however,  it may follow deliberate  or accidental ingestion or



result from absorption through broken skin (NAS,  1974).



     Much of the information on the harmful  effects of Cr(VI) is obtained from



occupational  exposures,  where the predominant effects are  on the respiratory



system and skin (NAS, 197*0.   As  with most information  derived in this fashion,



ejact knowledge about  length of exposure, concentration of  the  chemical, and



other variables are not known, making it  difficult to interpret the results or



assess any quantitative relationships between dose of chemical and its effect.








7.1.2.   Animal Studies.  Cr(III) compounds  have a very low  order of toxicity when



administered orally.   Oral LD_  values for the rat  have been reported as follows:



chromic chloride,  1.87 g/kg;  chromium acetate,  11.26  g/kg;  chromium  nitrate,



3.25 g/kg (Smyth et al.,  1969).  Kobayashi et al. (1976)  have determined oral



LD _ for chromium  trioxide in mice  and rats to  be 135 to  177 mg/kg and  80 to



114 mg/kg,  respectively.   Animals  died  over   a period  of  3  to  35 hours.
                                     7-1

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Symptomatology included diarrhea, cyanosis, tail necrosis, and  gastric  ulcer.



Surviving animals showed increases  in liver and testes weight  without  micro-



scopic changes.



     Cr(VI)  is more acutely toxic.  A primary effect of acute exposures is kidney



failure.   Oral administration  of  high  doses results in gastric  corrosion.   The




oral LD__ of sodium dichromate in humans has been reported as 50 mg/kg (NIOSH,




1979).



     Kidney effects are the  primary result of acute  exposures  to  chromium by



various routes.  Relevant studies are summarized in the following paragraphs.



     Mathur   et al.  (1977)  injected  rabbits   intraperitoneally with  2 mg/kg



chromium nitrate or potassium chrctnate daily for 3 or 6 weeks.   After 3 weeks of



exposure, kidneys from  animals dosed with chromium nitrate showed marked conges-



tion, extravasation of  red  blood cells in  the  intratubular spaces  and  tubular



necrosis.   Further  treatment,  up to  6 weeks,  did not produce  any additional



changes.



     The  kidneys from  animals given potassium  dichromate  showed marked conges-



tion and the walls  of the small blood vessels were thickened.   Glcmerular tufts



were  shrunken  in  some  places, while  proliferation  of  endothelial  cells,



obliterating the Bownan space,  was seen in others.   There  was necrosis  and



desquamation of the epithelium of the convoluted tubules.   Red blood cells were



found in the intertubular spaces.   Changes were similar whether animals  were



exposed for  3 or 6 weeks.



     Hunter  and Roberts (1933) dosed monkeys subcutaneously with 1 to 5 mfc of a



2% solution  of potassium dichrcmate.  Two monkeys were dosed every 3 weeks;  the



first dose was 1 mi, the second  dose was 2 mi, the third and  fourth doses were



4 mi, and the fifth dose was 5 mi. One monkey died following the final dose.  The
                                     7-2

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authors reported injury to the  proximal  and distal convoluted tubules and the




glcmeruli of the kidneys.



     Kirschbaum et al. (1981)  injected rats  subcutaneously with 20 mg/kg sodium



chronate.   Their  data demonstrate that  epithelial cell injury in  the kidney



occurred 2  to  4 hours  post-inject ion.  They postulate that the interaction of




chromium  with  specific  elements  of  the microfilamentous  system,  which  are



responsible for directing intracellular flow of reabsorbed solutes,  may be the



initial effect  of  chromium nephropathy.  Baines (1965) also reports renal damage



following exposure of rats to acute, subcutaneous doses of potassium  dichromate.



     Berry et al.  (1978) have  examined localization of chromium within the kid-



ney.    Rats  were  dosed by  intraperitoneal  injection  with  0.1 mg potassium



dichronate/100 g body weight.   Doses were given 5  times per week for 8 months.



Chromium was localized within  cells  of the proximal renal tubules, specifically



within lysosones.  Chromium was retained throughout  most of the  study period,



being eliminated only when necrosis  involved the entire cytoplasm of the tubule



cells.



     Evan and Dail  (197*0 also report data which indicate effects on the proximal



convoluted tubules following intraperitoneal administration of 10  or  20 mg/kg



sodium chromate and  the  formation of  large  lysosomal vacuoles in this region.



They  also  report  effects  on  mitochondria!  configuration  shortly following



exposure.



     Berndt (1976) reports in  vitro  data which indicate there may  be species



differences in  kidney susceptibility to chromium nephropathy.  In  kidney slices



from rats,  effects were independent of valence state.  Kidney slices from rabbits



were more sensitive  to transport process inhibition by Cr(VI).



     Mathur et  al. (1977) documented  effects  in other target  organs following



acute exposure  to  chromium.  Rabbits were dosed intraperitoneally  with 2 mg/kg
                                     7-3

-------
chromiun as chromium nitrate or as potassium dichronate.  Doses were given daily



and the animals were sacrificed after 3 or 6 weeks.  Administration of Cr(III)



for1  3 weeks  produced  changes  in the  brain,  including  occasional  neuronal



degeneration  in the cerebral cortex, marked chromatolysis, and nuclear changes



in the neurones.   Six weeks of exposure resulted in marked neuronal degeneration



in the  cerebral cortex accompanied by neuronophagia, neuroglial proliferation,



and meningeal congestion.




     Following  3  weeks   of  exposure  to Cr(VI) , congestion with perivascular



infiltration by inflammatory cells was noted.   In addition, some neurones in the



cortex  showed pyknotic  nuclei and dissolution  of  Nissl's substance.   Neuro-



nophagia and  focal  neuroglial  profileration  were also evident throughout  the



cerebral cortex.   Changes  following 6 weeks of exposure  were similar.   The



myocardium  of  animals exposed  to Cr( III) for  3 weeks  appeared to  be normal.



However, following 6 weeks  of  exposure, the myocardium showed marked congestion



and degeneration  of  muscle fibers.    Exposure  to Cr(VI) for  3 weeks  did  not



produce any abnormalities.   Exposure for  6 weeks produced changes  similar  to



those seen in animals exposed  to the  trlvalent salt.



     Tandon et al. (1978) report hepatic changes in rabbits exposed to chromiun.



Exposure conditions were  the sane as in the  previous study.   Exposure to Cr(III)



for 3 weeks produced marked congestion and dilation of the central  veins  and



sinusoids.   Discrete foci of  necrosis were  noted in  liver  parenchjima.   After 6



weeks, in addition to marked  congestion, extensive  hemorrhage  was  seen in the



parenchyma.   Slight nuclear  plecmorphism and mul tinucleated cells  were noted in



the lobules.  Bile duct  proliferation, increased cellularity and  proliferation



of fibroblasts around portal tracts were noted.



     Exposure to Cr(VI)  for 3 weeks  produced  more extensive  pathology than  did



Cr(III)  exposure.  The liver  capsule  was thickened and there was  marked conges
                                    . 7-4

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tion of central veins and adjacent sinusoids.  Large areas  of necrosis were seen



throughout the  parenchyma.   Changes  seen  following 6 weeks  of exposure were



similar to those described following  6 weeks of exposure to the  trivalent salt.



     In summary,  a  number of  organs  appear to be  targets  for acute chromium



toxicity.  Kidney  effects are the  best  docunented, because they  have received the



most intense study.
                                     7-5

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7.2  CHRONIC EFFECTS OF CHROMIUM EXPOSURE IN MAN AND ANIMALS
7.2.1.  Evaluation of the Carcinogenicity of Chromium

    The purpose of this section  is  to  provide an evaluation  of  the  likelihood
that chromium is a human carcinogen and,  on  the  assumption that it  is  a  human
carcinogen, to provide a basis  for  estimating its public  health impact
including a potency evaluation  in relation to other  carcinogens.  The
evaluation of carcinogenicity depends  heavily on animal bioassays and
epidemiologic evidence.  However, information on mutagenicity and metabolism,
particularly in relation to interaction with ONA, as well as the
pharmacokinetic behavior, have  an important  bearing  on  both  the qualitative
and quantitative assessment of  carcinogenicity.   The available  information on
these subjects is reviewed in other sections of  this document.   This section
presents an evaluation of the animal  bioassays,  the  human epidemiologic
evidence, the quantitative aspects  of  assessment, and finally,  a summary and
conclusions dealing with all  of the relevant aspects of carcinogenicity.

ANIMAL STUDIES
    A number of animal studies  have been  performed to determine whether  or not
chromium compounds are carcinogenic.   In  these studies  metallic chromium and
salts of both the hexavalent and trivalent states were  administered by various
routes.  The discussions that follow are  grouped by  these various routes of
administration.

Inhalation Studies
    Baetjer et al.  (1959)  exposed three strains  of  mice (strain  A,  Swiss,  and
C57B1) having high, medium,  and low spontaneous  lung  tumor incidences
respectively, to chromium-containing dust.   The  dust  was  similar to that  found
in the chromium chemical  manufacturing industry, containing 13.7% 003 and

                                     7-6

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6.9% Cr203 along with other metal oxides.  In addition to the chromium
compounds in the dust, I^C^Oy was added at a level  of 1.0%.   The
animals were exposed to the dust-laden atmosphere containing  between 0.5 and 1
mg of total chromium 4 hours/day, 5 days/week, for an average of 39.7 weeks
(range of 16 to 58 weeks).  Table 7-1     describes the specifics for each
exposure group.  At death or termination of exposure, the lungs were examined
using a low power microscope, and abnormal  tissues were submitted for
histologic confirmation of tumors.
    The incidence of lung tumors was not different in exposed mice of any
strain as compared with approximately equal numbers  of the appropriate strains
of unexposed mice of the same age.  There was also no difference in those
strains having high spontaneous tumor incidence in regard to  the average
number of tumors per mouse or the percent of mice with multiple tumors.   The
lung tumors present in both control  and treated animals were  adenomas, which
appeared histologically to be similar; however, in exposed animals, the
adenomas developed slightly earlier in the  strain A  mice.  Three additional
small groups of mice (two groups of 10 Swiss female  mice and  one group of nine
female strain A mice) were exposed to high  concentrations of  chromium dust
(7.8 to 13 mg Cr/m3) in a nose-only chamber 0.5 hours/day, 5  days/week,  for
43, 52, and 20 weeks, respectively.   Again, no increase in the incidence of
lung tumors was observed.
    In a lifetime chronic study, Nettesheim et al.  (1971) exposed 136 C57B1/6
mice of each sex 5 hours/day, 5 days/week,  to an  atmosphere containing
CaCr04 dust at a level  of 13 mg/m3,  with 95% of the  particles  less  than
0.6 u in size.  During the study, seven or  eight  mice were removed  for interim
sacrifice at 6, 12,  and 18 months; the sex  distribution and exact  number of
animals removed at each period  were  not stated.   At  autopsy,  sections  were

                                   7-7

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TABLE  7-1    .  INHALATION EXPOSURE OF MICE TO CHROMIUM-CONTAINING DUST
                         (Baetjer et al.  1959)
Strain
Swiss
Swiss
Swiss
Strain A
Strain A
Strain A
Strain A
C57B1
C57B1
Sex
F
M
F
F
F
F
M
M
F
Number of
Animals at
Start
11
10
127
34
45
110
52
50
61
Number of
Animals at
End
7
6
51
31
45
38
36
13
14
Duration of
Exposure
(weeks)
39
39
58
16
24
38
46
42
41
Estimate
of Total
Cr Inhaled
(mg of Cr)
520
520
692
141
210
359
406
667
589
                              7-8

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 taken  from  all major organs, and with the exception of the lungs, it was
 indicated that no  increase in tumor incidence was observed.  In the lungs,
 tumors  developed in six males and eight females, as compared with three males
 and  two females in the control group.  The tumors were described as
 alveologenic  adenomas and adenocarcinomas, although the number of each type of
 tumor  was not enumerated.  The authors claim in the discussion that CaCr04
 exposure resulted  in a significant increase in lung tumors; however, the
 performance of statistical analysis was not described in the results.  In a
 review  of this study, the International Agency for Research on Cancer (IARC
 1980) maintained that no excess in treatment-related tumors was observed.
 Because of  difficulties in determining the number and sex of the animals
 removed during the interim kills, it is impossible to perform independent
 statistical analysis of this data.
     Two other experimental groups in the aforementioned study were exposed to
 CaCrffy  dust following prior treatment with 100 R whole-body X-radiation or
 infection by influenza virus.  A slightly greater incidence of lung tumors was
 observed after combined exposure to influenza virus, as compared with exposure
 to CaCr04 alone.  As a result of exposure to two potential  cancer-producing
 agents,  these last two groups of animals cannot be used to provide any
 supporting evidence for the carcinogenicity of chromium.   With the limited
 experimental detail presented in this study, it is not possible to determine
 if the  small increase in lung tumors  represents a significant treatment-
 related  increase in tumor incidence.
     In the study described previously,  Baetjer et al.  (1959)  also  exposed 110
 (57 males and 53 females)  mixed-strain  rats (from Wistar  and  McCollum stock)
to the same chromium dust  to  which  the  mice were  exposed;  100 rats  (48 males
and 52 females)  of  similar age  distribution were  kept  as  controls.   The  level
                                       7-9

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of chromium in the air was 1 to 1.5 mg/m^ w-jth exposure again for 4
hours/day, 5 days/week, for > 70 weeks.  During the study,  nearly half of the
experimental  rats died, three developed lymphosarcomas involving the lungs,
while two additional  suspected lymphosarcomas involving the lungs were also
identified.  No rats developed bronchogenic carcinomas.  The authors
considered these findings suggestive of a chromium-induced  tumorigenic
response; however, since lymphosarcomas are common in these rats, the
experiment was repeated.  In the second study (Steffee and  Baetjer 1965),
Wistar rats were exposed to chromium-containing dust using  the same regime as
described previously.  Following autopsy, three alveologenic adenomas were
detected in the treated rats and two in the controls, and four lymphosarcomas
were present in both groups.  From this second study, it was concluded that
lymphosarcomas in rats are not associated with exposure to  chromium-containing
dust.
    Also, Steffee and Baetjer (1965) exposed eight rabbits  and 50 guinea  pigs
to mixed chromium dust containing 1.5 to 2.0 mg of total  chromium/m^ for  4
to 5 hours/day, 4 days/week, for 50 months and life, respectively.   The mixed
chromium dust exposure consisted of 2 days/week exposure to roast dust [as
described previously by Baetjer et al. (1959) supplemented  with 1.0% dry
K2Cr2°7 and tne m'ist of a 5-°?° section °f !<2Cr207» followed by a
1 day/week exposure to the mist of a 17.5% solution of ^CrO^, and a 1
day/week exposure to residue dust (roast dust from which Na2Cr04 was
extracted) supplemented with 1.0% dry I^C^Oy.  There were  no lung
tumors in the rabbits, and the incidence of lung tumors and other body tumors
was similar in the exposed and control guinea pigs.  Under  these experimental
conditions, inhalation of mixed chromium dust did not increase the incidence
of lung tumors.
                                     7-10

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    Laskin (1972) conducted a study in which rats (number and  strain  not
specified) were exposed by inhalation to air containing 2 mg/m^ calcium
chromate (VI).  The total  number of exposures were 589 over a  period  of R91
days.  The author reported one squamous cell carinoma of the lung  and larynx
and one malignant peritoneal  tumor.  Because of incomplete reporting  of the
experiment, this study is  considered to be inadequate to assess the
carcinogenicity of calcium chromate by inhalation.

Intratracheal Instillation Studies
    In further attempts to illustrate a lung tumorigenic response  from
chromium compounds, mixed chromium dust and chromium salts were instilled  into
the tracheas of experimental  animals.  Baetjer et al. (1959)  suspended  a
chromium dust, similar in composition to that used in the inhalation  studies,
in olive oil and zinc chromate and barium chromate in saline  prior to
intratracheal instillation into strain A, Swiss,  and C57B1  mice and mixed
breed rats (Wistar and McCollum stocks).  The mice each  received five to six
instillations of 0.01 to 0.05 mg of chromium at 4- to 6-week  intervals, while
the rats received 15 instillation  at the same dose at 2-week  intervals.  The
total  duration of the studies was  between 32 and  52 weeks.   The mice  treated
with chromium had a similar tumor  incidence as age-matched controls,  and the
rats in both the treated and  control groups had no benign or  malignant  tumors.
In continuing work, Steffee and Baetjer (1965) treated 62 strain A mice with
0.01 to 0.03 mg chromium by intratracheal  instillation of ZnCr04.   The
instillations were performed  at 2-week intervals  for a total  of six
injections, and the animals were observed until death.  The incidence of lung
adenomas was 31 of 62 in  the  treated animals and  7 of 18 in the untreated
controls.  In the controls treated with ZnC03, 3  of 12 animals  developed
                                    7-11

-------
lung adenomas.  The incidence of lung tumors in treated animals  was
statistically different from controls.  In the same study,  the instillation  of
chromium dust, ZnCrO^ and PbCr04 into the tracheas of guinea  pigs  (13 to
21 animals/group) and rabbits (7 to 10 animals/group)  produced no increase in
lung adenomas.  Hueper and Payne (1962) also reported  similar  negative results
after instillation of SrCr04 or CaCrO^ suspended in gelatin; however,  the
experimental detail in the report was insufficient for adequate  evaluation.
There is no convincing evidence that intratracheal  instillation  of  chromium
compounds results in the development of lung cancer.

Intrabronchial Implantation Studies
    Laskin et al. (1970) investigated the carcinogenic effects of chromium
compounds using the intrabronchial  pellet technique.   The compounds used  in
this investigation were:  chromic chromate, chromic oxide,  chromic  trioxide,
calcium chromate, and process residue.  Pellets were prepared  from  molten
mixtures of materials dispersed in  equal  quantities of cholesterol  carrier.
These studies included material  of  differing solubilities and  valences.   A
total  of 500 rats were under observation  for periods up to  136 weeks.   Lung
cancers that closely duplicate human pathology were found in these  studies
(Table   7-2   ).
    With the calcium chromate, eight cancers were found in  an  exposed  group  of
100 animals.  Six of these were squamous  cell  carcinomas and the other two
were adenocarcinomas.
  The National Institute for Occupational  Safety and Health (NIOSH  1975)
criteria document on chromium (VI)  described a written communication from L.S.
Levy in 1975 about an animal  study  done at Chester Beatty Research  Institute,
London.  Random-bred Parton Wistar  rats of both sexes  received a pellet in the
                                   7-12

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TABLE  7-2   .   CARCINOMAS PRODUCED WITH CHROMIUM COMPOUNDS IN RATS
                        (Laskin et al. 1970)
Material
Process Residue
Calcium Chromate
Chromic Chromate
Chromic Oxide
Chromic Tri oxide
Cholesterol Control
Number of
An i ma 1 s
100
100
100
98
100
24
Squamous Cells Adeno- Heptocellular
Carcinoma carcinoma Carcinoma
1 1
6 2 1
1

2

                              7-13

-------
left inferior bronchiolus via tracheotomy under anesthesia.   The rats were
kept for 2 years.  One hundred rats were set up for each of  the
chromium-containing-material  test groups.  The pellets which were implanted
contained 2 mg of test material  suspended 50/50 (w/w)  in cholesterol.
Negative control  groups received either blank metal  pellets  or pellets and
vehicle.  Positive control  groups received 3-methylcholanthrene.  Lungs of all
rats either dying during the study or killed at its termination were examined
both macroscopically and microscopically.  Apart from  those  in the lung,
tumors were similar both in type and number in all  groups.   The bronchial
tumors found and  microscopically confirmed are given in  Table  7-3    along
with the average  induction  periods.  Additional  lung tumors, not of bronchial
origin and not considered by the authors to be causally  related to
implantations are also listed in Table  7~3   .   The majority of bronchial
tumors were large keratinizing squamous cell  carcinomas.  Intrathoracic
invasions, particularly to  the right lung in the hilar region, were common and
metastases to local lymph nodes  and to kidneys were seen.
    Squamous cell carcinomas were found in 8/100 (  P < 0.05) rats receiving
calcium chromate, 3/100 rats receiving zinc chromate (zinc potassium
chromate), 3/100  rats receiving  chromic chromate dispersed in silica, and
1/100 rats receiving ground chromic acid.  It may be that the chromic acid
implantation produced a carcinoma only because it was  converted to a
less-soluble chromium (VI)  material by reaction  with cholesterol.  Because of
its extremely great oxidizing ability, some of it may  be have been chemically
reduced by cholesterol, forming  chromic chromate.  Calcium chromate produced
carcinomas in 5/100 (P < 0.05) rats when mixed with primene, and carcinomas in
7/100 (P < 0.05)  rats when  mixed with diphenylguanadine. Primene 81-R
benzoate and diphenylguanidine failed to produce tumors  when administered  by
                                   7-14

-------
       TABLE  7-3   .  LUNG TUMORS FOUND AND MICROSCOPICALLY CONFIRMED
                    [Levy and Venitt (1975), NIOSH (1975)]
Experi-
mental Corn-
Group  pound
No.    No.    Test Material
No.    Bronchial   Induction
Rats   Carcinoma   Period
in     of Left    in Days
Group  Lung       (Range)
Lung Tumors
not Associated
with Treatment
 1      1     Ground chro-    100
              mite ore

 2      2     Bolton high
              lime residue      "

 3      3     Residue after     "
              alumina pre-
              cipitation

 4      4     Residue from      "
              slurry tank-
              free of
              soluble Cr

 5      5     Residue from      "
              vanadium
              filter

 6      6     Residue from    101
              slurry
              disposal tank
              Sodium dichro-  100
              mate dihydrate
                               Pulmonary aden-
                                oma of left lung
                               Anaplastic car-
                                cinoma of upper
                                left lung
                               Adenoma of right
                                lung
                               Fibrosarcoma of
                                upper left lung
8
9

10
lit
12

8
9

10
11
12

Sodium chromate "
Chromic acid "
(ground)
Chromic oxide "
Calcium chromate "
Chromic chloride "
hexahydrate
ii
1

0
8
0


560


604(473-734) P<0.05
Lymphoma of
right lung
    *Zinc potassium cnromate.
    tP-value is calculated using the Fisher Exact Test (one-tail).
                                             (continued on the following page)
                                  7-15

-------
                      TABLE    7-3 .   (continued)
Experl
mental
Group
No.
13

14
21

22t
23

t24

25
26

15

16

28


20
17
18
19
27
Com-
pound
No.
13

14
15A

15B
16A

16B

17A
17B

15

16

28


20
17
18
19
27
No.
Rats
in
Test Material Group
Zinc chromate- "
type II*
Chrome tan "
Diphenyl- "
guanidine (DPG)
DPG + calcium "
Primene 81-R 100
benzoate
Primene + cal- "
cium chromate
Chromic chromate "
Chromic chromate "
dispersed in silica
Pellet + chol- 150
estorol
Blank Pellet

Pellet + chol- 100
estorol +
Kieselguhr
100% 3-MCA 48
100% 3-MCa
50% 3-MCA
25% 3-MCA
50% 3-MCA 50
Bronchial
Carcinoma
of Left
Lung
3

0
H

7
0

5

0
3

0

II

»


34
36
18
13
36
Induction
Period
in Days
(Range)
708(657-734)




656(502-732)


620(440-732)


698(666-730)








493(217-730)
498(270-701)
474(284-696)
517(297-698)
498(269-732)
Lung Tumors
not Associated
with Treatment





P<0.05


P<0.05




Adenoma of
right lung
Adenocarcinoma
of right lung








*ZTnc potassium cnromate.
tP-value is calculated using the Fisher
Exact Test (one-tail),
                             7-16

-------
 themselves.  No bronchial carcinomas were found in negative control groups or
 in  rats receiving sodium dichromate dihydrate or sodium chromate.

 Intrapleural Injection Studies
     Production of pulmonary tumors by the intrapleural injection of chromium
 compounds has also been attempted.  Hueper and Payne (Hueper 1955, Hueper
 1958,  Payne 1960a, Hueper and Payne 1962, Hueper 1961) described a series of
 studies in rats treated by intrapleural injection of a number of hexavalent or
 trivalent chromium compounds.  Powdered metallic chromium was injected into
 the  pleural cavity of rats, guinea pigs, and mice under the dose schedule
 described in Table 7-4     (Hueper 1955).  No significant increase in tumor
 incidence, either at the injection site or in other organs, was observed.
 Payne  (1960a) implanted chromite roast, from which the soluble NajjCrC^ was
 extracted, into the pleural  cavity of 35 rats.  Each rat received 25 mg of
 this material plus 50 mg of sheep fat, which corresponded to 2 mg of total
 chromium, of which 0.4 was hexavalent chromium.  None of the 35 control
 animals developed tumors, while three of the treated animals developed
 implantation site tumors.  In an earlier study (Hueper 1958) using chromite
 roast not leached of Na2Cr04, none of the 25 treated male Bethesda rats
 developed implantation site  tumors during 24 months; however,  the early death
 of nine of the  treated animals appreciably decreased the animals at risk.  No
 implantation site tumors  were observed within 24 months in 42  rats by Hueper
and Payne (1962)  following eight implantations of 25 mg of trivalent  chromium
acetate in gelatin over a 13-month period.   Using a  single implant of 2 mg of
K2Cr2°7 1nto tne  Plural  cavity, 1 of  39 rats developed a tumor at the
implantation  site.  In this  study,  only CaCr04 elicited a high  tumor
incidence  following  implantation.   Of  the  14  rats  treated with  12.5 mg of
                                    7-17

-------
                         TABLE  7-4   .  EXPOSURE SCHEDULE FOR BIOASSAY OF CHROMIUM COMPOUNDS BY INTRAPLEURAL INJECTION
Species
mice
mice
rats
rats
— rats
CO
rats
rats
rats
rats
guinea pigs
Strain
C57B1
Strain A
Bethesda Black
Bethesda Black
Bethesda Black
Bethesda Black
Bethesda Black
Osborne-Mendel
Osborne-Mendel
NR*
Number of Animals
50
55
25
35
42
39
14
25
25
26
Compound
metallic
chromium powder
mixed chromium dust
chromlte roast
chromlte roast
minus Na2CrO^
chromium acetate
K2Cr207
CaCr04
metallic
chromium powder
chromlte ore
metallic
chromium powder
Compound (mg)
0.001
1 or 2
25
25
25
2
12.5
16.8
36.7
67.2
Number of Injections
6 injections at 2
week intervals
4 Injections at 4
to 6 week Intervals
single Implant
single Implant
1n 50 mg of fat
8 Implantations over
13 months
single Implant
single Implant
6 Injections at
monthly Intervals
6 Injections at
monthly Intervals
6 injections at
monthly Intervals
Reference
Hueper 1955
Baetjer et


al. 1959
Hueper 1958
Payne 1960a
Hueper and
Hueper and
Hueper and
Hueper 1955
Hueper 1955
Hueper 1955

Payne 1962
Payne 1962
Payne 1962



*NR = Not reported.

-------
 CaCr04»  eight  developed  tumors  at the  site of  implantation.  These studies
 suggest  that intrapleural  implantation of some hexavalent chromium compounds
 might be carcinogenic, with  CaCr04  producing the most dramatic response.
 Hueper (1961)  reports further evidence that many hexavalent chromium compounds
 produce  tumors upon  intrapleural implantation, while trivalent compounds are
 less  effective;  however,  no  experimental detail, including dose, was provided.
 A summary of the tumor incidences reported are presented in Table  'J-H" .
     Baetjer et al.  (1959)  also  used intrapleural injection to assess the
 carcinogenicity of mixed chromium dust,  containing both trivalent and
 hexavalent chromium, in  male (30 animals) and  female (25 animals) strain A
 mice.  The mice received four doses of dust suspended in olive oil, with each
 dose  containing 0.07 mg  of chromium (Table 7-5    ).  No increase in tumor
 incidence or number  of lung  tumors  per mouse was observed during the period
 extending 52 weeks after the first  treatment.  Davis (1972) injected trivalent
 chromite [FeO(CrAl)203l  into the pleura! cavity of 25 BALB/c mice and
 observed only  small  granulomas  in the  lungs.   The animals were treated with a
 single injection of  5 mg of  finely  ground ore  (
-------
     TABLE  7-5   .  COMPOUNDS REPORTED TO HAVE BEEN TESTED FOR
            CARCINOGENICITY BY INTRAPLEURAL IMPLANTATION*
                            (Hueper 1961)
Compound
Calcium chromate
Sintered calcium chromate
Strontium chromate
Lead chromate
Barium chromate
Sodium di chromate
Zinc yellow
Chromic chromate
Chromite roast
residue
Chromium acetate
Sheep fat controls
Valence
+6
+6
+6
+6
+6
+6
+6
+6, +3
+6, +3
+3
NA§
Number of Rats
with Tumorst
n
20
17
17
3
1
0*
22
26
5
1
0
Percent
57
49
74
9
3
0
63
74
14
3
0
tThere were 35 rats per group at the start.
§NA = not applicable.
                                  7-20

-------
nor a greater number of tumors per tumor-bearing  lung  than  the  72 control
mice.  More recently, Stoner et al. (1976)  and Shirnkin et al.  (1978)  reported
Similar negative results for trivalent 02(504)3  following
intraperitoneal  administration to strain A  mice.   In this study,  groups  of  20
mice received 100, 50, or 20% of the maximum tolerated dose (2,400 mg/kg) in
24 injections given three times per week.  The animals were examined  for
tumors at the end of 30 weeks.  In all of these studies,  positive controls
were used to demonstrate the sensitivity of the strain A mouse  to the
development of chemically-induced multiple  lung tumors.
    Although increased risk of lung cancer  has been associated  with the
chromium industry as discussed below, it has proven difficult  to  demonstrate a
carcinogenic response in the lungs of experimental  animals.  Trivalent
chromium compounds have not produced lung tumors  after inhalation,
intratracheal implantation, or intrapleural implantation, while hexavalent
chromium was not carcinogenic by inhalation or intratracheal  instillation.
Some hexavalent chromium compounds did produce tumors  following intrabronchial
or intrapleural  implantation; however, the  small  number of  animals (14)  used
by Hueper and Payne  (1962) in the study of  CaCrO/^ and  the lack  of detail in
the report of Hueper (1961), where a number of hexavalent chromium compounds
were reported to be carcinogenic, make it difficult to evaluate the
carcinogenicity of these compounds to rodent respiratory tissue.   For these
reasons, the experimental studies of respiratory cancer in  animal studies do
not provide substantial confirmation for the industrial observation.   However,
the limited data does suggest that of the two valences, hexavalent chromium is
more likely to be the etiologic agent in chromium-induced cancer.
                                     7-21

-------
 Injection  Studies For Sites Other Than Lung
     Attempts have been made to demonstrate chromium-induced carcinogenesis in
 other  than  respiratory tissue.   In an early study, Hueper (1955) injected
 either powdered chromium or chromite ore into the marrow cavity of the femur
 of  rats, rabbits, and dogs.  The experimental conditions are described in
 Table   7-6   .  Of the animals treated, only one rat developed a tumor at the
 site of injection, and other tumors observed in the treated and control groups
 were not considered treatment-related.  Similar negative results were obtained
 by  Hueper  (1955) following intraperitoneal  injection of chromium powder in
 rats and mice and intravenous administration of chromium powder in mice, rats,
 and  rabbits.  The experimental conditions used in these studies are also
 presented in Table  7-6  .  Although some tumors were present in treated rats,
 these  tumors were similar to those in the controls except for two treated rats
 that developed unique insulomas of the pancreas after intraperitoneal
 administration of powdered chromium, and two treated rats that developed lung
 adenomas.  While none were observed in the  controls, the causal  relationship
 between chromium exposure and these tumors  is highly questionable.
     Injection site tumors developed in animals after subcutaneous
 administration of chromium compounds in some, but not all,  studies.  Payne
 (1960a) treated groups of 52 C57B1  mice (26 males and 26 females) with size
 fractionated particles of chromium residue  dust or chromic  phosphate.   The
 animals received a single subcutaneous injection of 10 mg,  after which they
 were observed for life.   The composition of the dust fraction as related to
 trivalent and hexavalent  chromium is presented in Table  7-7   .  There was a
 low incidence of injection site tumors (3 of 52)  observed in animals  treated
with the unfractioned  residue  dust,  while no tumors were present in the
 controls or animals  treated  with smaller particles, even though  these  smaller
particles  had a higher proportion of hexavalent  chromium.   In  a  study  of
                                    7-22

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TABLE   7-6   .  EXPERIMENTAL CONDITIONS  USED  TO  STUDY  THE  EFFECT  OF  INTRAFFMORAL,  INTRAPERITONEAL,  AND  INTRAVENOUS  ADMINISTRATION  OF  CHROMIUM
                                                               (Hueper  1955)
Route
intrafemoral
Intrafemoral
Intrafemoral
Intrafemoral
"71 Intrafemoral
Intrafemoral
intraperitoneal
intraperltoneal
intravenous
intravenous
intravenous
Species
rats
rats
rabbits
dogs
rats
rabbits
mice
rats
mice
rats
rabbits
Strain
Osborne-Mendel
Wistar
Dutch
Mixed breed
Osborne-Mendel
Dutch
C57B1
Histar
C57B1
Wistar
NR*
Number of Animals
(M, F)
25 M
25 M
8 F
5 F
15 M, 10 F
4 F
50 M
25 M
25 M
25 M
8 F
Chromium
Compound
powdered chromium
powdered chromium
powdered chromium
powdered chromium
chromite ore
44% Cr203
chromite ore
44% Cr203
powdered chromium
powdered chromium
powdered chromium
powdered chromium
powdered chromium
Dose
100 mg
100 mg
140 mg
170 to 399 mg
followed in 24
months by 340
to 798 mg
36 mg
147 mg
1 mg/week
for 4 weeks
5 mg/week
for 6 weeks
0.25 mg/week
for 6 weeks
9 mg/week
for 6 weeks
2.5 mg/kg/week
for 6 weeks,
treatment repeated
4 months 1 ater
Duration of
Observation
24 months
24 months
3 to 58 months
60 months
24 months
20 to 50 months
21 months
NR*
18 months
NR*
3fi months

-------
TABLE  7-7   .  LEVELS OF HEXAVALENT CHROMIUM IN
            FRACTIONATED RESIDUE DUST
                  (Payne 1960a)
Material
                   Weight of Chromium as Cr/dose
            Hexavalent (mg)                 Total  (nig)
Vehicle
 (tricaprylin)

Dust residue extracted
 with H20

Dust residue
 5 to 10

Dust residue
 < 2

Chromic phosphate
                                 0


                               0.037


                               0.17


                               0.45

                               0.003
                                               0


                                             0.50


                                             0.69


                                             0.68

                                             2.64
                        7-24

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Identical  design,  Payne (1960b)  treated  mice  with  sintered  003,  sintered



CaCr03, and CaCrO^   Only one injection  site  tumor was  observed,  and this



was in an animal  treated with CaCr04.   Roe  and  Carter  (1969)  reported that



20 weekly injections of CaCr04 at  a  dose of 5 mg for the  first  2  weeks and



0.5 mg for the remaining 18 weeks  resulted  in a 75% (18 of  24)  incidence of



injection site tumors.   Although the title  of Roe  and  Carter's  (1969) article



and the legend to the tabulated results  described  the  route of  administration



as subcutaneous injection, the experimental section and the conclusion



describe the treatment as an intramuscular  injection into the flank.  As a



result of this uncertainty, it is  unclear whether  any  chromium  compound has



been demonstrated to produce injection site tumors following  subcutaneous



administration.








Implantation Studies



    Intramuscular implantation has also been used  with varying  success to



demonstrate that chromium compounds  are carcinogenic.   Hueper (1958) and Payne



(1960a) implanted chromite roast or  chromium residue dust mixed with sheep  fat



into the thigh of Bethesda black rats.  The respective incidence  of injection



site tumors was 3 of 31 for animals  given 25 mg of chromite roast and 1 of  35



for animals receiving the same dose  of dust.  In both  studies,  no tumors were



present in the sheep fat-treated control animals.   Payne  (1960a)  also



implanted 10 mg of chromium dust in  the thigh of 52 C57B1 mice  and observed  no



injection site tumors.  Hueper and Payne (1959) and Payne (1960b) used similar



techniques in the investigation of pure chromium compound.   In  a  small study,



Payne  (1960b) observed two injection site tumors in six Bethesda  rats after



implantation of a gelatin capsule  containing 12.5  mg of CaCrO^  In the



study by Hueper and Payne (1959),  25 mg of  a chromium  compound  was mixed with
                                   7-25

-------
sheep fat prior to implantation  into  groups  of 35 Bethesda black rats.  The
implantation site tumor  incidence  was  8  of 35 for CaCrO^ 8 of 35 for
Sintered CaCrO/^, 15 of  35 for  Cr03, and  0 of 35 for BaCrO^  On
implantation of 10 mg of sintered  CaCr04 into the thighs of 52 C57B1 mice,
nine implantation site  tumors  developed, however, only one tumor developed
following the implantation of  CaCrO^.  Hueper (1961)  reported on the
development of implantation site tumors  following treatment of rats with a
number of chromium compounds (Table    7-8  ); however, there was no
experimental detail in  this report, including dose given, and thus, it  is
difficult to relate this study to  the  other  reports of implantation site
tumors.  The intramuscular implantation  technique has provided relatively
consistent findings that some  hexavalent chromium compounds are tumorigenic in
laboratory animals.
    Neither trivalent nor metallic chromium  compounds have produced
implantation site tumors following intramuscular implantation.  Hueper  and
Payne (1962) implanted  25 mg of  chromic  acetate into  the thigh of 35 Bethesda
black rats.  After a 24-month  observation period, only one animal developed an
injection site tumor.   Using powdered chromium, Sunderman et al.  (1974)
observed no tumors after 112 weeks in 24 male Fisher  rats.  Atomic absorption
spectroscopy indicated  that each rat  received 2 mg of dust.  Similar results
were obtained by Hueper (1955) following repeated injection of powdered
chromium into the thigh of 25  C57B1 mice.  Each animal received two injections
of 0.1 mg at 2-week intervals, and this  was  repeated  3 weeks later for  a total
dose of 0.4 mg.  No tumors developed  in  the  15 mice that survived 3 to  13
months.  Although trivalent chromium  was not tumorigenic following
intramuscular implantation, only one  compound was tested under a  limited
experimental protocol,  and it  is only speculative that zero valent and
                                   7-26

-------
TABLE   7-8   .  COMPOUNDS REPORTED TO HAVE BEEN TESTED FOR CARCINOGENICITY
                       BY INTRAMUSCULAR IMPLANTATION
                               (Hueper 1961)
Compound
Calcium chromate
Sintered calcium
chromate
Strontium chromate
Lead chromate
Barium chromate
Sodium di chromate
Zinc yellow
Chromic chromate
Chromite roast
residue
Chromium acetate
Sheep fat control
Valence
+6
+6
+6
+6
+6
+6
+6
+6, +3
+6, +3
+3
NAt
Number of Tumors*
9
12
15
1
0
0
16
24
1
1
0
Percent
25
34
43
3
0
0
46
69
3
3
0
   *Tnere were 35 rats/group at tne start.
   tNA = Not applicable.
                                    7-27

-------
trivalent chromium would continue  to  give  negative  results with further
testing.

Oral Studies
    Trivalent chromium has been tested  for carcinogenicity by  the  oral route
in mice and rats.  Schroeder et al.  (1964) exposed  a  group of  108  (equal
numbers of male and females) Swiss mice to drinking water containing  5 ppm  of
chromium as chromium acetate.  The lifetime exposure  to  this level  of chromium
had no effect on longevity of females,  and only  a  slight decrease  in  longevity
in males.  There was no increase in the tumor incidence  in the treated animals
as compared with controls.  In a similar study,  Schroeder et al.  (1965)
exposed 46 male and 50 female Long Evans rats to drinking water containing  5
ppm of chromium as chromium acetate.   Again, lifetime exposure to  this level
of chromium had only slight effect on longevity, with no increase  in  tumors in
treated as compared to control animals.  It should  be noted that  only one dose
level of chromium was used in this study,  and from  the lack of overt  signs  of
toxicity, it may be concluded that higher dose levels could be tolerated.
Higher dose levels would increase the likelihood of detecting  a carcinogenic
response from a weak carcinogen.  Lane and Mass  (1977) observed carcinogenic
activities of chromium carbaryl as well as synergistic activities  with
benzo(a)pyrene in rats following tracheal  grafting  techniques.
     Ivankovic and Preussman (1975) incorporated  0^03 into the diets  of  60
male and female BD rats.  The 0^03  was baked into  bread at levels of 1,
2, or 5%, and fed to the rats 5 days/week for 2  years.  The only  effect  of
treatment was a dose-dependent decrease in liver and  spleen weight.  Both the
longevity and tumor incidence in the  treated animals  were  similar  to  that of
control animals.  Again, the lack of  major toxic effects of treatment may
                                   7-28

-------
 indicate that this chromium compound could have been tested at higher levels
 in the diet.  The author's commented that the negative may have resulted from
 the poor absorption of chromium from the gastrointestinal tract.

 Summary of Animal Studies
    A summary of the animal carcinogenicity studies of chromium is presented
 in Table  7-9    .  At present, it has not been successful to induce tumors in
 laboratory animals following exposure by inhalation and ingestion routes.
 Both trivalent and hexavalent chromium have proven ineffective in producing
 lung tumors by inhalation.  Similar negative results have been obtained
 following the ingestion of trivalent chromium; however, only low doses were
 used and hexavalent chromium has not been tested.  There is some evidence that
 chromium, particularly some hexavalent chromium compounds, are carcinogenic
 following subcutaneous injection or intrabronchial, intrapleural, or
 intramuscular implantation; however, implantation site tumors have only
 consistently been demonstrated using intramuscular implantation.  Of all  the
 chromium salts, calcium chromate is the only one which has been found to be
 carcinogenic in rats after intrabronchial and intramuscular implantations.
 Calcium chromate, strontium chromate, and zinc chromate can produce local
 sarcomas in rats at the site of application.  Although the studies available
 indicate that metallic chromium powder and trivalent chromium are not
 carcinogenic, these compounds have been studied less extensively than
 hexavalent chromium.   The relevance of studies using intramuscular
 implantation to human  risk following inhalation or oral  exposure to chromium
compounds is not clear;  however,  these animal  studies may indicate that
hexavalent chromium is  more likely to be  the etiologic agent  in human
chromium-related cancer.   Using the International  Agency for  Research on
                                   7-29

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                                        TABLE  7-9    .  CARCINOGENICITY OF CHROMIUM COMPOUNDS  IN  EXPERIMENTAL  ANIMALS
VjJ
O
Route of
Administration Compound
inhalation chromium containing
dust


Inhalation chromium containing
dust


inhalation chromium containing
dust
inhalation CaCr04 dust






inhalation chromium containing
dust




Inhalation chromium containing
dust
Inhalation chromium containing
mist and dust


Species/Strain
mice/Strain A



mice/Swiss



m1ce/C57Bl

m1ce/C57B1






rats/mixed breed
Wlstar and McCollum




rats/Wistar

rabbits
guinea pigs


Dose Duration
as Chromium of Exposure
0.5 to 1 mg/m3 4 h/d, 5 d/wk
for 16 to 54 wk


0.5 to 1 mg/m3 4 h/d, 5 d/wk
for 39 to 58 wk


0.5 to 1 mg/m3 4 h/d, 5 d/wk
for 41 to 42 wk
4.33 mg/m3 5 h/d, 5 d/wk
for life





1 to 1.5 mg/m3 4 h/d, 5 d/wk
for > 70 wk




1 to 1.5 mg/m3 4 h/d, 5 d/wk
for life
1 to 5 to 2 mg/m3 4 to 5 h/d, 4 d/wk
for the life of
guinea pig or 50
months for rabbits
Findings
No increase in the
incidence of lung
tumors or number of
tumors /lung
No Increase in the
incidence of lung
tumors or number of
tumors/lung
No lung tumors
observed
6 of 136 males and 8
of 136 females
developed lung tumors
as compared with 3 of
136 females and 2 of 136
male controls; the
significance is not clear
Increased incidence of
lymphosarcomas
involving the lungs, 3
of 100 in experimental
and 0 of 85 in control
group
No change 1n lung
tumor Incidence
No increase in the
incidence of lung
tumors

Reference
Baetjer et al .
1959


Baetjer et al .
1959


Baetjer et al.
1959
Netteshelm
et al. 1971





Baetjer et al .
1959




Steffee and
Baetjer 1965
Steffee and
Baetjer 1965


                                                                                                                           (continued  on  the  following page)

-------
TABLE  7-9
(continued)
Route of
Administration
Intratracheal
Intratracheal
Intratracheal
Instillation
Intratracheal
Instillation
Intratracheal
1ntniat1on
Intrapleural
Injection
Intrapleural
Injection
Intrapleural
Injection
Intrapleural
injection
Intrapleural
injection
Compound
chromium dust or
BaCrO-4 OP ZnCr04
ZnCro4
chromium
dust
ZnCrOa or
PbCr04
ZnCr04 or
PbCr04
Cr powdered metal
mixed chromium dust
FeO(CrAl)203
chromlte ore
Cr powdered metal
Species/Strain
mice/Swiss A
Swiss, C57B1
mice/Strain A
rats /mixed breed
Hi star and McCollum
rabbit
guinea pigs
m1ce/C57Bl
mice/Strain A
m1ce/Ba1b/c
rats/Osborne Mendel
rats/Osborne Mendel
guinea pigs
Dose
as Chromium
0.01 to 0.05 mg/
Injection
0.01 to 0.03 mg/
injection
0.02 ing/injection
2.3 to 2.8 mg/
Injection
0.7 to 0.86 mg/
0.001/mg
0.07 mg/1nject1on
1 mg
36.7 mg (of ore)
16.8 mg
67.2 mg
Duration
of Exposure
5 to 6 injections
at 4 to 6 wk
Intervals
6 Injections at
2 wk intervals
15 injections at
2 wk Intervals
3 to 5 Injections
at 3 mo Intervals
6 Injections at
6 wk intervals
6 Injections at
2 wk intervals
4 injections at
4 to 6 wk Intervals
single injection
6 Injections at
1 mo Intervals
6 injections at
1 mo intervals
Findings
No Increase in the
incidence of lung
tumors or the number
of tumors/lung
No statistical Increase
1n lung tumors, 31
No lung tumors
observed
No lung tumors
observed
No Increase 1n lung
tumor incidence
No significant Increase
1n Injection site
tumors
No Increase in lung
tumor Incidence or
number of lung tumors/
mouse
Only small
granulomas observed
No significant Increase
1n injection site
No significant increase
in injection site tumors
Reference
Baetjer et al .
1965
Steffee and
Baetjer 1965
Baetjer et al.
1959
Steffee and
Baetjer 1965
Steffee and
Baetjer 1965
Hueper 1955
Baetjer et al .
1959
Davis 1972
Hueper 1955
Hueper 1955
                                                          (continued on the following page)

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TABLE 7-9
(continued)
Route of
Administration
intrapleural
implant
Intrapleural
Implant
Intrapleural
Implant
Intrapleural
implant
Intrapleural
Implant
Intrabronchial
Intrafemoral
Intrafemoral
Compound
chromlte roast
minus Na2CrO$
K2Cr207
CaCr04
Cr(C2H302)3
chromlte roast
variety of
chromium compounds
metallic chromium
chromlte ore
Species/Strain
rats/Bethesda Black
rats/Bethesda Black
rats/Bethesda Black
rats/Bethesda Black
rats/Bethesda Black
rats/Parton
Wistar
rats/Osborne-Mendel
rats/Wistar
rabbits/Dutch
rats/Osborne-Mendel
rats/Dutch
Hose
as Chromium
25 mg (of roast)
0.35 mg
4.2 mg
5.2 mg/implant
25 mg (of roast)
2 mg
100 mg
100 mg
140 mg
15 mg
64 mg
Duration
of Exposure
single Implant in
sheep fat
single Implant 1n
in sheep fat
single Implant in
1n sheep fat
8 implants over
13 mo
single Implant
single Implantation
single injection
single Injection
Findings
3 of 35 animals
developed implant
site tumors; none
in controls
1 of 39 animals
developed implant
site tumors; none
in controls
8 of 14 animals
developed implant
site tumors, none
In controls
No Implantation site
tumors 1n 24 animals
No implant site
tumors
See Table
No injection site
tumors developed
except for a single
tumor 1n one rat
No Injection site
tumors
Reference
Payne 1960a
Hueper
and Payne 1962
Heuper
and Payne 1962
Hueper
and Payne 1962
Hueper 1958
Levy and
Venitt 1975
as reported
1n NIOSH
(1975)
Hueper 1955
Hueper 1955
                                                         (continued on the following page)

-------
TABLE  7-9  .   (continued)
Route of
Administration Compound
Intrafemoral metallic chromium
1ntraper1toneal 02(804)3
intraperltoneal metallic chromium
1ntraper1tonea1 metallic chromium
Intravenous chromlte ore
Intravenous metallic chromium
Intravenous metallic chromium
Species/Strain
dogs/mixed breed
mice/Strain A
mice/C57Bl
rats/W1star
mice/Strain A
mice/C57Bl
rats/Wistar
rabbi ts/NR
nose
as Chromium
170 to 399 mg
f ol 1 owed by
340 to 798 ng
1.3 to 6.6 mg/
Injection
1 mg /Injection
5 mg/1nject1on
1.95 to 3 mg
0.25 mg/1njection
9.0 mg/inject1on
2.5 mg/kg
Duration
of Exposure
second treatment
given 24 mo
after first
24 Injections given
3/wk
4 Injections at
1 wk intervals
6 Injections at
1 wk intervals
single injection
6 injections at
1 wk intervals
6 injections at
weekly intervals
repeated in 4 mo
Findings
No injection site
tumors
No increase in lung
tumor incidence or
number of lung
tumors/mouse
No tumors observed
Pulmonary adenomas in
2 of 25 animals; none
in controls
No Increase in lung
tumor incidence or
number of lung tumors/
mouse
No tumors in mice
while tumors in rats
were identical to
controls
No adverse effects
after 36 months
Reference
Hueper 1955
Stoner et
al. 1976;
Shimkin et al .
1978
Hueper 1955
Hueper 1955
Shimkin and
Leiter 1940
Hueper 1955
Hueper 1955

-------
                                                                TABLE 7-9    .  (continued)
—i
 i
Route of
Administration
subcutaneous








Intramuscular


Intramuscular

Compound Species/Strain
chromium residue dust m1ce/C57B1
chromium residue dust,
5 to 10 p
chromium residue dust,
< 2 p
chromic phosphate
sinstered 003
sinstered CaCrO.3
CaCr04
metallic chromium mice/C57Bl


CaCr04 mice/C57Bl
sinstered CaCr04
Dose
. as Chromium
0.5 mg
0.69 mg

0.68 mg

2.64 mg
0.52 mg
0.37 mg
0.33 mg
0.1 mg


3.3 mg
3.3 mg
Duration
of Exposure
single injection








2 Injections at
2 wk intervals
repeated 3 wks later
single Implant

Findings Reference
3 of 52 animals Payne 1960a,b
receiving chromium
residue dust and 1 of
52 receiving CaCr04
developed injection
site tumors



No tumors in 25 Hueper 1955
animals

9 of 52 mice treated Payne 1960b
with sinstered CaCrO^
   Intramuscular


   intramuscular


   intramuscular
chromium residue dust   m1ce/C57Bl
metallic chromium
Cr(C2H302)3
rats/Fisher
                      10 mg  (of dust)
2 mg
rats/Bethesda  Black    5.2 mg
single implant


single Injection


single Implant
had implantation site
tumors, while 1 of 52
treated with CaCr04
developed tumors

No injection site
tumors developed

No tumors 1n 24
animals

1 Implantation site
tumors in 35
treated animals
                                                                        Payne 1960a
Sunderman et
al. 1974

Hueper
and Payne 1962

-------
Cancer (IARC) classification scheme,  this level  of evidence in rats  would be
sufficient for concluding that calcium chromate  and some relatively  insoluble
hexavalent chromium compounds are carcinogenic in animals.
                                  7-35

-------
 EPIDEMIOLOGIC STUDIES
 Chromate  Production
    The early association of respiratory cancer with employment in the
 chromate  industry has been reviewed by Baetjer (1950a).  The first case report
 appeared  in 1911, with a total of 122 reports of respiratory cancer in
 chromate  workers collected between this date and 1950.  These cases were
 predominantly in German and American industries, with only one case reported
 in Scotland in 1890, and no cases reported in England or France.  The workers
 in the chromate industry were exposed to chromite ore (trivalent chromium) and
 sodium monochromate, sodium bichromate, and chromic acid (hexavalent
 chromium), along with other vapors and gases associated with the chromate
 manufacturing process.  The early German investigators suggested that
 hexavalent chromium was the etiologic agent in respiratory cancer, since
 respiratory cancer was not associated with the mining of trivalent chromite
 ore.  These early observations do not provide information on the relative
 incidence of respiratory cancer in the chromate workers as compared with the
 general population, nor were the studies sufficiently large or controlled to
 support the conclusion that chromium exposure was related to respiratory
 cancer.   However, these early reports were the impetus for initiating a number
 of epidemiologic studies of the chromate manufacturing industry in the United
 States, Great Britian, and Japan.  Although a relatively large number of
 epidemiologic studies have been conducted of this industry, the individual
 studies were often analyzing cancer incidence from the same cohort of workers.
 In order to clarify the interrelationship of the  cohorts in these  studies,  the
 location of the plants from which each study derived its exposed population is
presented in Table  7-10   .   It should be made clear that studies  of the same
plant  by different investigators  often resulted in  the vital  statistics of
                                7-36

-------
                 TABLE  7-10
                                       LOCATION OF CHROMATE MANUFACTURING PLANTS WHICH PARTICIPATE!! IN EPIOEMIOLORIC STUDIES AMI PLANTS FROM
                                                        WHICH VITAL STATISTICS WERE OBTAINED FOR EACH STUDY
Location
  of
 Plant
                 Machle
                  and
                firegorlus
                 1948
Brlnton
et al.
1952
(also pub.
as part of
PHS 1953)
Mancuso
 and
Hueper  Mancuso
 1951    1975
Baetjer
 1950a
Hayes
et al .
1979
 H111
  and
Ferguson
 1979
Taylor
 1966
Enterllne
  1974
Bldstrup
 1951
Bldstrup
  and
Case 1956
Alderson
 et al .
 1981
         Hatanabe
Ohsakl    and                  Korallus
et al.   Fukuchl    Satoh et    et al 1.
1978      1975      al. 1981     198Z
filen Falls.
 NY
Vjo
-~j
Jersey City, NJ
 Plant »1    +
 Plant 12    +

Baltimore,
 MD          +

Kearny, NJ   +

Newark, NJ   +

Palnes-
 vllle, OH   +

Bolton
 England

Rutherglen
 England

Eaglescllff
 England
                                                                                               +t
                                                                                                                                                                    wing page)
                                                                                                                                             (continued on the
    + » Participating chromate manufacturing plants.
    - = Non-participating chromate manufacturing plants.
    * = The plant in the Watanabe and Fukuchl (1978) and the OhsaM  et al. (1978) studies may be one 1n the same, 1t Is Impossible to tell from the
literature.  Both plants were reported to be located on Hokkaido Island, Japan, however.
    t - Plant #1 1n Jersey City, New Jersey 1s the larger of the two plants.  Machle and r.regorius (1948) reported that It had 350 employees compared
to 150 employees 1n Plant 12.

-------
                                                                      Table  7-10   .   (continued)
                             BMnton
                             et al.
                              , ,                                            H1"                                                                     Watanabe
         Location    and       (also pub.   and                      Hayes     and                                      Bldstrup   Alderson   Ohsakl      and                Korallus
           of      Gregorlus   as part of  Hueper  Mancuso  Baetjer  et al.  Ferguson  Taylor  Enterllne    Bldstrup      and       et al.    et al.    Fukuchl   Satoh et    et al.
          Plant      1948       PHS  1953)    1951     1975     1950a   1979     1979      1966     1974        1951       Case 1956   1981      1978       1975     al. 1981     1982


         Hokkaido
          Islands
 I        Tokyo,
*•*>        Japan
CO
         Leverkusen
           W.  Germany --           ---.._.

         Verdingen,
         W. Germany  --           ---.___


            *• -  Participating chromate manufacturing plants.                                                   ~~	~~—"	~~	
            - =  Non-participating chromate manufacturing plants.
            * =  The plant In the Watanabe and Fukuchl (1978) and the Ohsakl et al. (1975) studies may be one 1n the same, 1t 1s Impossible to tell from the
         literature.  Both plants were reported to be located on Hokkaido Island, Japan, however.
            t =  Plant »1 1n Jersey City, New Jersey 1s the larger of the two plants.  Machle and Gregorlus (1948) reported that 1t had 350 employees compared
         to 150 employees 1n Plant 12.

-------
individual workers being used in more than one study.  The result was that in
many cases, these epidemiologic studies verified the observations of previous
studies rather than adding evidence from additonally exposed population
groups.
    In response to the early reports associating lung cancer with the chromate
industry, the industry initiated a retrospective epidemiologic study of the
seven chromate plants in the United States (Machle and Gregorius 1948).  A
total of 1,445 workers were employed in the seven plants, with each plant
employing between 50 and 500 workers.  The company group life insurance
records were used to determine cause of death.  Adequate records were
available for six of the plants, and the cohort of men actively working in the
chromate industry consisted of 11,019 man-years of experience.  In this
cohort, 156 deaths were observed, 32 of which were from lung cancer.  An
additional 10 lung cancer deaths were reported of the 37 deaths from the plant
with work records unsatisfactory for the purpose of the epidemiologic
analysis.  The period of study varied with each plant from 4 to 17 years,
depending on the availability of mortality data.   Of the total deaths observed
in the chromate industry, 21.8% (42 of 193) were from lung cancer compared to
an expected 1.3% (10 of 733) as calculated from a comparable industrial  group
not exposed to chromium (industrial  life insurance policyholders for the year
1947, Metropolitan Life Insurance Co.).  This is statistically significant at
P < 0.01.  When examined individually, five of the seven plants were reported
to have respiratory cancer proportionate mortality ratios from 13 to 31 times
that expected.   The crude respiratory cancer moraltity rate per 1,000 males
was also found  to be significantly (P < 0.01) increased over the crude lung
cancer mortality rate of the group of life insurance holders.   This  was  true
for both the age group 50 and under  and the age group 50 and over.   A slightly
                                  7-39

-------
 increased crude mortality rate from digestive system cancer was also reported
 (1.18 per 1,000 versus 0.59 per 1,000,P <0.01).
    Estimates of exposure were not made in this study, since analytical data
 were not available for a large portion of the period studied, and work records
 did not report the shifting of personnel to different positions in the plant
 during the course of employment.  A limitation to this study is that no age
 adjustment was done in the comparison of the mortality rates of the control
 group with the mortality rates of the exposed group.  In this regard, however,
 it should be noted that there was a dramatic difference in lung cancer
 mortality between the chromate workers and the control  group for both the 50
 and under age category and the over 50 age category.  Thus, it is unlikely
 that a preponderance of older persons among the chromate workers was
 responsible for the large difference in lung cancer mortality between the
 chromate workers and the control  group.  Another limitation is that the
 authors used a lung cancer mortality rate for oil  refinery workers in 1933-38
 for comparison with the chromate workers of 1930-47.  Lung cancer increased
 dramatically in U.S. males between 1930 and 1947 (about five times).  Thus, it
 is possible that the lung cancer mortality rate for the control  group may have
 a lower rate than what would have been found for the period 1930-1947.
 However, the difference in lung cancer between the controls and the chromate
 workers is so dramatically different, it is unlikely that this difference
 could be explained by selection of a control  group from a slightly different
 time period than that of the exposed group.
    Brinton et al. (1952) used the disability records between January 1,  1946
 and December 31, 1950 of the seven United States chromate plants to determine
occupational  diseases associated  with the chromate industry.   These data  were
 subsequently published as part of the Public  Health Service (1953) report,
                                7-40

-------
 "Health of Workers in a Chromate-Producing Industry."  The cohort was limited
 to men who belonged to the company disability plan, which prior to 1949 was
 voluntary.  After 1949, the participation was mandatory in all  but two plants,
 although participation in these plants was near 100% by the end of the study.
 Only deaths that occurred within 1 year of beginning disability status were
 included in this study, and the determination of the cause of death was made
 solely on what was listed as "cause of death" on the death certificate.  The
 health experience of a "large group of industrial  workers (predominantly
 white)" was used for comparison.  The incidence of most disabilities was
 comparable between chromate workers and the reference population with the
 exception of cancer.  Cancer at all sites had an incidence of 7.1 per 1000 in
 chromate workers and 0.7 per 1000 in the control populaiton (P < 0.01);
 however, it should be pointed out that the actual  number of individuals who
 developed cancer was small (26 individuals) in the group of chromate workers.
 Brinton et al. (1952) further studied the cancer experience in  the chromate
 industry using death records obtained from the group disability insurance plan
 between the years 1940 and 1950.  During this time, information was available
 from two plants, while one plant had records from  1943 to 1950, three plants
 from 1946 to 1950, and one plant from 1949 to 1950.  During this 11-year
 period, 44 deaths from cancer at all  sites and 32  deaths from respiratory
 cancer were observed.  This corresponds to a 4.5-fold increase  in cancer at
 all sites and  a nearly 29-fold increase in respiratory cancer in the chromate
 workers as compared to males in the United States.  There was an indication in
the data that  the increased risk of respiratory cancer was greater in nonwhite
 as compared to white chromate workers; however, the number of individuals was
 small  when the cohort of chromate workers  was subjected to this further
subdivision.   The authors  commented on the limitations the study placed on
                                    7-41

-------
the make-up of the cohort as a result of the use of death records obtained
solely from the company disability plan and suggested that the observed
increased  risk of respiratory cancer represents a minimal value.
    Baetjer (1950a, b) studied a cohort consisting of lung cancer patients in
two Baltimore hospitals to determine if employees of the local chromate plant
were overrepresented in this group.  The records from Johns Hopkins Hospital
were examined from 1925 to 1946, and from Baltimore City Hospital from 1930 to
1948; 198  and 92 confirmed cases of lung cancer in males were obtained,
respectively, from each institution.  Control  groups consisted of age-matched
male patients with a hospital stay of >10 days chosen at random from the
hospital records.  An additional control was used for Johns Hopkins; this
consisted  of patients with cholelithiasis, since patients with this disease,
as well as lung cancer patients, may preferentially select a large medical
facility.  At Johns Hopkins, 7 of 198 lung cancer patients had worked in the
chromate industry; none of the 226 randomly sampled controls and none of the
177 cholelithiasis controls had worked in the  chromate industry.  At the
Baltimore  City hospital, 4 of 92 lung cancer patients and 0 of 499 control
patients reported exposure to chromium compounds.  The percentage of chromate
workers in the lung cancer group was statistically significantly (P < 0.01)
greater than expected.   If one were to calculate the unadjusted odds ratios of
having lung cancer and being exposed to chromium, the odds ratios would be
about 32 for the Johns Hopkins cases and about 23 for the Baltimore City
Hospital  cases.   Both are statistically significant (P < 0.05).
    Mancuso and  Hueper (1951) used the vital statistics  found for all
employees who worked for >^ 1  year in the Painesille,  Ohio chromate plant  during
1931 to 1949 to  investigate chromate production-associated lung cancer.  Of
the 2,931 deaths of  males in the county in which the plant was located, 34
                                 7-42

-------
(1.2%) were from respiratory cancer, while of the 33 deaths among the chromate
workers, 6 (18.2%) were from respiratory cancer.   This  difference is
significant at P < 0.01.  Mancuso and Hueper (1951)  indicated that 95% of the
workers were exposed exclusively to insoluble chromite.  Chemical analysis of
the organs of two deceased workers, one who had died from lung cancer and one
who had died from bladder cancer, revealed that the  lungs appeared to be the
major storage depot for chromium, with approximately 390 and 250 ug of
chromium per 10 g of tissue detected in each individual respectively.  The
high level of chromium in the first subject, the one who had died from lung
cancer, was still detected despite the fact that the individual  had not been
exposed to chromium for a period of 3.4 years.   Chromium levels  measured in
the lungs of nonexposed individuals were nearly zero.  Mancuso and Hueper
(1951) suggest that the presence of insoluble chromium  in the lung may be an
etiologic factor in the observed higher incidence of cancer; however, the
present data are too limited to support this conclusion.
    Mancuso (1975) followed the vital  status until  1974 of 332 chromate plant
workers from his earlier study (Mancuso and Hueper 1951) who were employed
from 1931-1937.  Over 50% of the 332 employees  in this  cohort had died by
1974.  Mancuso divided this group of workers into those employed from 1931-32,
1933-34, and 1935-37.  Mancuso found that 63.6, 62.5, and 58.3% of the cancer
deaths in the 1931-32, 1933-34, and 1935-37 groups,  respectively, were lung
cancer deaths.  The latency period for the lung cancer  deaths was found to
cluster around 27-36 years.  The author found that lung cancer deaths were
dose-related to insoluble (trivalent), soluble  (hexavalent), and total
chromium exposure, and thus, the author concluded that  lung cancer mortality
was associated with both trivalent and hexavalent chromium.  Because workers
in this study were exposed to both trivalent and  hexavalent chromium, and as
                                      7-43

-------
exposure to one increased so did  exposure  to  the  other,  an  observed
dose-response to trivalent chromium may merely  be a  reflection of  a
dose-response to hexavalent chromium or vice-versa.   Thus,  it  is questionable
whether the author's conclusion with regard to  a  dose-response to  both
hexavalent and trivalent chromium is correct.  Furthermore, the  lung  cancer
death rates, which purport to show this dose-response,  are  based on very  small
numbers, and thus the finding of  dose-response  is probably  questionable.   For
six deaths due to lung cancer, chromium levels  well  above control  values  were
found in the lungs as long as 15  months to 16 years  and 3 months after  last
exposure to chromium suggesting that the lung retains chromium for a
considerable period of time.
    More recently, Taylor (1966)  studied a group  of  1,212 chromate workers for
24 years using Old-Age and Survivors Disability Insurance  records. All  of
these workers were employed in three United States chromate plants (Baltimore,
Maryland; Painesville, Ohio; and  the larger of  the two plants  in Jersey City,
New Jersey) for 2.3 months during  the 4-year period of 1937  to  1940.   The  vital
statistics of the group were obtained through 1960,  and where  deaths  occurred,
the cause was determined from death certificates.  Causes of death in this
cohort were compared with age and cause-specific  mortality  rates  for  civilian
males in the United States.  The  most dramatic  increase in  the standardized
mortality ratios  (SMR) for chromate workers occurred in respiratory cancer,
with excesses of 8.5 times (71 observed and 8.3 expected)  observed by the
termination of the study in 1960.  Also, the length  of experience  in  the
chromate industry, used as the only indication  of the extent of  exposure, was
compared with the incidence of lung cancer in this cohort.   After 1937,
duration of employment was determined from insurance records,  while prior to
this date, duration of employment was estimated by extrapolation from the
                                  7-44

-------
age-specific employment  experience  of  the  cohort.  Using this method to
determine length of exposure,  Taylor found that respiratory cancer mortality
showed a dose-response by length of time exposed to chromate.
    Enterline (1974) recalculated the  expected deaths for the study by Taylor
(1966) but provided no explanation  as  to why this  was done.  Enterline
calculated a respiratory cancer SMR of 942 for the 20-year observation period
1940-1960.  Taylor had calculated a respiratory cancer  SMR of 850 for the
period 1937-1960.  Enterline (1974) showed that the relative risk of
respiratory cancer was greater in workers  in the  age  group <45 years  (14.1
times) as compared with the older age  group  of  55 to  64 (6.8 times).
Enterline also showed that the risk of respiratory cancer'was highest  shortly
after the cohort was identified, suggesting  "a  short  latent period  which  is
probably the result of exposure to a  very potent  carcinogen."
    Hayes et al. (1979) conducted a cohort mortality  study of workers  at  a
Baltimore, Maryland chromium chemical  production  plant.  The  plant  underwent
extensive changes in the mill  and roast operation and bichromate operations  in
1950 and 1951 when a new facility was built to house these  operations and in
the chromic acid and special products operations  in 1960 when a  new facility
was built to house these operations.  The new facilities were constucted  for
the purpose of lowering employee exposure to chromium.   In  this  study, vital
status was determined on newly hired workers between 1945 and 1974 who had at
least 90 days of employment.  This cohort consisted of 2,101 employees, 1,803
laborers, and 298 managers, of which vital  statistics were obtained for 88% of
the group as of  1977.  A comparison of lung cancer in this cohort was made
using SMRs from  the cause-specific mortality rates of Baltimore males and
tested for significance using Poisson probability distribution.   Workers were
divided  into two exposure groups; the high  or questionable exposure group
                                      7-44a

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consisted of employees who worked in the old facilities and workers of unknown
exposure, and the low exposure group consisting of workers employed only in
the new facilities.  In this study, no information was available on the actual
levels of exposure or the extent of difference in exposure between the new and
old plants.  Analysis of lung cancer mortality by specific jobs was done after
matching lung cancer cases by race, age, time of initial  employment, and
duration of employment to employees that died from non-cancer causes.
    The SMR for lung cancer in the entire cohort of hourly workers was 202
(95% confidence limits of 155 to 263), which was statistically significant
(P < 0.01).  The SMRs for both short (90 days to 2 years) and long (>3 years)
term workers of the high and low exposure group are presented in Table 7-11 .
There was an apparent dose-response relationship, as associated with length of
employment, for the group initially hired 1950-59.  The lung cancer SMR was
statistically significant (95% confidence limits did not  include 1.00) for the
workers with "high and questionable exposure" employed >3 years, and who had
initially been employed 1950-59.  In analyzing lung cancer by job description,
only workers in the special  products department or employees who worked in
both the special products and bichromate departments, the so called "wet-end"
of the production process (the production process in the  mill and roast
department is referred to as the "dry end"), showed a significantly (P < 0.05)
elevated relative risk of lung cancer of 2.6 and 3.3, respectively.
    In a study of the same plant investigated by Hayes et al. (1979),  Hill  and
Ferguson (1979) used a novel statistical analysis in an attempt to demonstrate
any trends in the risk of lung cancer associated with the modernization of the
plant.  The statistical  method used was "probability window analysis"  which
provided a method of comparing the number of cases of lung cancer in any equal
time period.  In comparing the number of lung cancers prior to 1951 (the time
                                  7-45

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    TABLE   7-11  .  OBSERVED NUMBER OF DEATHS,  STANDARDIZED MORTALITY RATIOS (SMRs),  AND 95% CONFIDENCE LIMITS (95% CL)
                                FOR DEATHS DUE  TO CANCER OF THE TRACHEA,  BRONCHUS,  AND LUNG
                       AND THE NUMBER OF REPORTED DEATHS FOR WHICH NO CERTIFICATE COULD BE OBTAINED,
                       BY YEAR OF INITIAL EMPLOYMENT,  EXPOSURE CATEGORY AND TOTAL DURATION EMPLOYED,
                                      FOR WORKERS INITIALLY HIRED AS HOURLY EMPLOYEES
                                                    (Hayes et al. 1979)
Duration of
Employmentt
  Cause of
  Death
   Low Exposure

   Observed
No. of Deaths
                                                                  Exposure Category*
SMR (95% CL)§
Questionable and- High Exposure


              SMR (95% CL)§
   Observed
No.  of Deaths
Short
Long
Trachea, bronchus    NA
 and lung

Cause not            NA
 determinedll

Trachea, bronchus    NA
 and lung

Cause not            NA
 determined'!
                   INITIALLY HIRED 1945  to  1949

                          NA


                          NA


                          NA


                          NA
                                       20


                                       25


                                       13
                  1.8
                  (1.1 to 2.7)

                  NA
                  3.0
                  (1.6 to 5.2)

                  NA
    *Based upon whether work exposure was exclusively in a new facility.   See text.
    tDuration of employment, short (90 days to 2 years), long (>3 years).
    §Calculated using an assumption that the observed number of deaths  is  distributed as  a  Poisson  random
variable, P = 0.025, in each tail.
    ^ICause not determined are those reported deceased for whom no death certificate  could be obtained.   If these
are distributed by cause of death in a similar way as the known deaths  (cancer of the trachea,  bronchus,
and lung 15%) the reported SMRs would he increased slightly.
    NA = Not applicable.
                                                                                           (continued  on  the  following  page)

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                                                TABLE  7-11   .   (continued)


Duration of
Employmentt

Short

Long


All


Low Exposure
Cause of Observed
Death No. of Deaths

Trachea, bronchus 2
and lung
Cause not 3
determinedfl
Trachea, bronchus 3
and lung
Cause not 0
determinedfl

Trachea, bronchus 0
and, lung
Cause not 0
determinedfl
Exposure Category*
Questionable and High Exposure
Observed
SMR(95% CL)<) No. of Deaths SMR (95% CL)§
INITIALLY HIRED 1950 to 1959
0.7 (0.1 to 2.6) 12 1.8
(0.9 to 3.1)
NA 7 NA
4.0 (0.8 to 11.7) 9 3.4
(1.6 to 6.5)
NA 0 NA
INITIALLY HIRED 1960 to 1974
NA 0 NA
NA 0 NA
    *Based upon whether work exposure  was  exclusively  in  a  new  facility.   See text.
    tDuration of employment; short  (90 days  to  2  years),  long  (X3 years).
    §Calculated using an assumption  that the observed  number of deaths  is  distributed as a Poisson random
variable, P = 0.025, in each tail.
    HCause not determined are those  reported deceased  for whom  no death certificate could be obtained.   If these
are distributed by cause of death in a similar  way  as  the known deaths  (cancer of the trachea, bronchus,
and lung 15%) the reported SMRs  would  be increased  slightly.
    NA = Not applicable.

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of start up of the new bichromate plant) with those after 1951, Hill  and
Ferguson (1979) report a statistically significant (P < 0.01)  decline from 23
to 7, and the authors suggested that this reduced risk of lung cancer resulted
from the engineering improvements that took place in 1951.  Comparing  the time
periods 1932-41, 1942-51, 1952-61, and 1962-71 the number of bronchogenic
carcinoma deaths falling within these time periods ("windows") are 9, 7, 1,
and 0, respectively.  The authors found a significant (P <  0.01)  difference
when comparing the four groups under the null  hypothesis that  all  classes have
equal probabilities of having cancer mortality.  The Carcinogen Assessment
Group would agree with the review of this study by IARC (1980) in  which  they
stated that no conclusion on improved safety in this chromate  plant could be
made, since the analysis only compared lung cancer cases and not  rates of lung
cancer.  Also, this analysis does not allow for the long latency  period
generally associated with lung cancer in the chromate industry.
    Chromate plants in both England and Japan have also been studied  to
determine if there is an association between employment in  this industry and
lung cancer.  In an early survey, Bidstrup (1951) performed lung  X-rays  of 724
workers employed in the three chromate plants in England in 1949.   This  survey
detected only one case of lung cancer, while the expected number  would have
been 0.4, as indicated from the mass radiography units of the  Ministry of
Health.  Of the 724 workers examined, 237 were employed for M5 years, and
although the numbers were too small  for definite conclusions,  Bidstrup (1951)
suggested that it was unlikely that a 25-fold increased risk of lung  cancer,
as reported in studies of United States plants, was associated with employment
in the British plants.  Bidstrup's study was a cross-sectional  study  of  only
currently employed workers, however.  Workers  with lung cancer or  with
symptoms of lung cancer have probably dropped out of the working  population.
                                       7-48

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Thus, the results are likely to be underestimates  of  what  the  difference  in
the incidence or the risk of lung cancer in  this group  of  workers  would be.
    Bidstrup and Case (1956) performed a follow-up study between  1949  and  1955
on the 723 workers who participated in the radiographic survey.   During this
time, 217 workers were lost to the study as  a result  of change in  employment.
Of the remaining men, all but 59 were alive  in 1955 when the  study was
terminated.  The age and cause specific deaths of  these chromate  workers  were
compared with the number expected based on the vital  statistics for the male
population of England and Wales.  There was  no difference  in  either death
rates for neoplasms other than lung cancer or deaths  from  other causes;
however, for lung cancer, there were 12 observed  deaths with  only  3.3
expected.  This increase of 360% was statistically significant (P  = 0.005).
There was also a trend for greater risk in the age group  <45  years (7  observed
and 1.3 expected); however, the numbers were too  small  to  demonstrate  if  this
trend was statistically significant.  The effects  of  place of residence,
social class, and smoking habits on lung cancer incidence  were considered  too
small to account for this 360% increase in risk.   The authors note that this
study was of short duration and continued follow-up of this cohort would
probably give rise to an even greater increased risk  of lung  cancer.  Of
interest in this study is the fact that the authors reported  that 217  workers
were lost to follow-up because of change in employment.  Change of employment
may well indicate a change in health status  and thus  may  suggest  that  the
difference in lung cancer mortality again may be  underestimated.
    Alderson et al. (1981) conducted a cohort mortality study of  workers  at
three chromate plants in Great Britain.  This was  a follow-up of  the earlier
studies by Bidstrup (1951) and Bidstrup and  Case  (1956).   Subjects were
eligible for this study if they had had an X-ray  examination  at work,  had
                                    7-49

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worked for a minimum of one year's  continuous  service  and  were  employed  from
1948 to 1977.  Two of the plants, those at  Bolton  and  Rutherglen  were  closed
in 1966 and 1967, respectively.   Following  the closing of  these two plants,
production became concentrated at the Eaglescliffe plant.   The  national
mortality rates for England and  Wales were  used to calculate expected
mortality for the plants at Bolton  and Rutherglen.  For all  plants  together
the observed lung cancer mortality  to that  expected was statistically
significant (observed/expected = 2.419, P < 0.001).  For the individual
plants, the observed/expected ratio was not significant (observed/expected  =
1.00 with 5 observed and 4.98 expected P <  0.44) at the Bolton  plant while  it
was significant (P < 0.001) at the  Eaglescliffe (observed/expected  = 2.156
with 36 observed and 16.20 expected) and Rutherglen (observed/expected = 2.854
with 75 observed and 26.28 expected) plants.  It should be noted  however that
the cohort at the Bolton plant was  relatively  small (number = 202)  and not
large enough to be reasonably able  to detect a difference  between the  observed
and the expected lung cancer deaths.  It should also be noted that  the
observed to expected nasal cancer mortality was statistically significant
(P < 0.05) at the Rutherglen plant.  The authors were  interested  in
determining if plant modifications  affected the relative risk of  disease in
men in the earlier study by Bidstrup and Case  (1956).   The authors  found that
the observed/expected ratio of lung cancer deaths decreased from  3.0
(P < 0.01) for those who worked before the plant modifications, to  2.0
(P < 0.005) for those who worked before and after the modifications, to 1.9
(P = 0.290) for those who worked after the modifications were completed.
    Because of the confounding factors of age  distribution of the workforce,
duration of employment, duration of follow-up, and the influence  of
environment, the authors did a multivariate analysis of the data.  For each
                                  7-50

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individual, the analysis compared the risk of developing lung cancer based on



the following factors:  duration of employment, duration of follow-up,



calendar period of employment, factory age at entry, and estimated degree of



chromate exposure.  The authors found the greatest contribution to the risk of



lung cancer mortality to be that of duration of employment followed by



duration of follow-up, calendar period of employment, entry age, and estimated



degree of chromate exposure.  In this study the authors reported that an



earlier report of the study cohort (case unpublished) presented data on the



smoking habits of the workers for 70 percent of the workers who had completed



a questionnaire.  Results from this survey found that the percentage of heavy



smokers was lower for the study cohort than that reported for England and



Wales (Todd 1962).  Thus, the authors concluded that there was no evidence



from the respondents to the questionnaire of a major risk of lung cancer due



to smoking compared to the population as a whole.  Such a conclusion must be



considered questionable, however, considering that 30% of the cohort did not



respond.  Other studies have found that the percentage of smokers among



nonrespondents to questionnaires is higher than among respondents (noil and



Hill 1964, Criqui et al 1979.



    Watanabe and Fukuchi (1975) followed a group of 136 chromate production



workers who worked or had worked in a chromate production factory on Hokkaido



Island, Japan.  The criteria for inclusion in the cohort was that one must be



exposed for more than 9 years to chromium compounds.  The follow-up period was



14 years, from 1960 to 1973.  Eight cases of lung cancer and two possible



cases of lung cancer were identified during the follow-up period.  The author



did not state how the ten cases were identified.  Seven of these ten cases had



died during the observation period, including the two questionable cases.  The



date and cause of the deaths were ascertained by their death certificates
                                     7-51

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and/or hospital  records.   Based  on  national  vital  statistics  data,  the
expected number  of lung cancer deaths  for  this  population  would  be  0.33.   Thus
the observed number of lung cancer  cases  is  21.2 times  greater  (if  the  2
questionable cases are counted with the observed lung cancer  death)  or  15.2
times greater (if only the five  certain cases of lung cancer  are counted).
Both observed numbers are significantly  (P < 0.05)  greater than  expected,
however.
    Ohsaki  et al. (1978)  studied the incidence  of  lung  cancer in a  cohort  of
67 active chromate workers and 487  retired chromate workers at  a factory on
Hokkaido Island  in Japan.  It  is not known at this  time whether  this  is the
same factory as  was in the Watanabe (1975) study.   The  average  exposure was
22.8 years (10 to 36 years), with 133  workers exposed for  > 10 years.   In  this
population, the  authors diagnosed 10 patients with  lung cancer,  and  determined
from death records that an additional  four cases had occurred.   The  incidence
of lung cancer in these chromate workers  was 658 per 100,000  compared to 13.3
per 100,000 for  the Japanese population.   (It is presumed  that  the  latter  rate
is for the male  population, although the  authors did not state  this
explicitly).  No correction was  made in this study  for  smoking  habits, even
though all  but two of the individuals  with lung cancer  were heavy smokers.
    Sano and Mitohara (1978) reported  that of 36 deceased  chromate  workers in
Tokyo, 19 died of cancer of the  respiratory  organs. Although this  report
merely related case histories, the  authors maintained that it supports the
risk of cancer associated with the  chromate  industry.   Of  particular  interest
in this report were the total  metal  analyses of two workers exposed  to
chromium who died of respiratory cancer.   Along with excessive  levels of Cr,
there were elevated levels of  Ni, Co,  Be,  V, and Mn in  the lungs, indicating
exposure to other possible carcinogens.
                                    7-52

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    Satoh et al. (1981) conducted a mortality and morbidity study of 896
workers engaged in the manufacture of chromium compounds for one or more years
during the period 1918 to 1975 at a plant in Tokyo, Japan.  It was reported
that during the period 1934 to 1975, 84% of the chromium compounds
manufactured were hexavalent and 16% were trivalent.  The plant was closed in
1975; workers were followed until 1978 or until death.  Data on the cause and
date of death were collected from death certificates or other "reliable
written testimony."  In addition to the 896 workers that were followed, there
were 165 chromium workers who were not included in the study due to a lack of
"necessary information."  All of these were retired workers whose registered
permanent residence and current status were unknown and whose vital status as
of the end of 1978 could not be determined.  Of these 165, approximately 80%
had left chromium work prior to 1949, and for 65% the date of birth was
unknown.  The average number of years as a chromium worker for the 165 not
included in the study was about 7 years, less than the 10-year average for the
896 workers that were included in the study.  The authors analyzed the
mortality data by four different time periods, 1918-49, 1950-59, 1960-69,
1970-78, and the overall  time period 1918-78.  They reported the observed and
expected number of deaths and SMR for various diseases and found that there
was an excess risk of lung cancer for each of the time periods (mortality
ratio = 9.5 for the 1918-78 time period, P < 0.005).  No excess risk  of death
from any other disease was found for any of the time periods or for the
overall  time period.   Satoh et al.  also analyzed the data by length of working
experience (1-10 years, 11-20 years, and 21+ years) for the time periods
1950-1959, 1960-1969,  and 1970-1978.  For lung cancer deaths,  but not for
deaths from "all  other cancers,"  the ratio of observed to expected deaths
increased by length  of working experience.
                                        7-53

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    Satoh et al.  studied morbidity  by  examining  health  insurance  records for



the period 1974 through 1977 to determine  if  any sickness occurred  in the 81



chromium-exposed workers as compared with  the 82 nonexposed workers  in the



plant during that period.  The author  reported that  although the  numbers were



small, there was no apparent difference between  the  exposed and the  nonexposed



groups in the number of cases for any  major disease  category.   It should be



noted that respiratory cancer and perforation of the septum are compensate



and thus do not appear in the health insurance records.   Three years after the



end of the chromium exposure, all 94 workers  who had been exposed for  1 to 28



years (average 14.9 years) were given  a complete series  of liver  and kidney



function tests.  All values were reported  to  fall within the  normal  range



characteristic of Japanese males.



    Korallus et al. (1982) reported the lung  cancer  SMRs for  two  West  German



chromate-producing plants during the period  1948-1979.   The population of



North Rhineland-Westphalia was used as the comparison group  in  calculating the



SMRs.  The respiratory cancer SMRs for both  plants,  Leverkusen  and  Uerdingen,



1.92 and 2.24 respectively, were both statistically  significant  (P  < 0.05)  for



the period of the study.  The authors  reported that  when the  study  period  was



divided  into six four-year intervals  (1948-52, 1953-57, 1958-62,  1963-67,



1968-72, 1973-77) and  one two-year interval   (1978-79) that  the  respiratory



cancer SMRs for these  intervals  generally decreased  in both  plants  over  the



period of the study.   This decrease was rather inconsistent  in  the  Uerdingen



plant, however.  Additionally, three  subcohorts  were defined:  members with



beginning of exposure  prior to January 1, 1948  (Group I), members exposed



before and after the  "change  in  manufacture"  (the change in  manufacture  began



in  1948  and was completed  in  1957 at  Leverkusen  and in 1963 at Uerdingen)



(Group  II), and members  with  beginning of exposure and after either "change in
                                    7-54

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manufacture" or before this provided at least half the exposure time was after
completion of the changeover (Group III).  The "change in manufacture"
represents the incorporation of the "no-lime" processing of the chrome ore
which is believed to result in a reduction of the carcinogen risk.  For both
plants there was a clear drop in the lung cancer SMR from Group I to Group
III.  For Uerdingen, the lung cancer SMRs were 2.76, 2.60, and 0.96 for Groups
I,  II, and III.  For Leverkusen, the corresponding lung cancer SMRs were 2.85,
1.97, and 0.54.  In neither the Leverkusen plant nor the Uerdingen plant,
however, were the SMRs for Groups I, II, or III significantly different
(P  < 0.05) from each other.  Thus, the decrease in the respiratory cancer SMR
from Groups I to III might be due to chance.  A reduction in nasal
perforations, symptomatic of chromium exposure, was also observed for Groups I
to  III.  This latter finding is statistically significant at P < 0.05 (chi
square test for linear trend in proportions) in both plants and would support
a conclusion of a decrease in chromium exposure from Group I to Group III.

Chrome Pigment Industry
    Two studies have been conducted on the mortality in the chrome pigment
industry in which workers were exposed only to hexavalent chromium.  Langard
and Norseth (1975)  reported on three pigment plants in Norway that were in
operation between 1948 and 1972.  One of the plants was brought on line only
in 1972, however, the year the study ended.  Between these dates, 133 workers
were identified as  being employed at the three plants.  Of the 133, 24 had
been employed > 3 years, and of this cohort, six cases of cancer, three of
which were lung cancer and one which was gastrointestinal  cancer, were
identified through  the Cancer Registry of Norway.   All  three of the lung
cancer cases  had  been employed 5 years or longer.   Two other cancer cases, one
                                  7-55

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with prostate cancer and the other with  nasal  cancer were  identified  among the
133 workers.  These cases did not  qualify  for  membership in the  cohort
employed for > 3 years, however.   Data  from the  Cancer  Registry  indicated an
expected number of lung cancer cases  among those employed  of  0.079; thus, the
observed number of cases was 38 times that expected.   Exposure levels as
determined by personal  monitoring  were  reported  for the plants for the year
1972, the year in which the study  ended, with  chromium  levels in the  two older
plants ranging between  0.04 and to 1.35 mg/m^, and levels  in  the new  plant
between 0.01 and 0.08 mg/m3.  The  distribution of the  number  of  employees
per year was not presented, although  it  was indicated that only  seven or eight
employees worked in the plants between  1948 and  1950,  with this  number
increasing slowly to a  level of 30 workers by  1972.  Although an increased
risk of lung cancer was indicated, two  of  the  individuals  with lung cancer
were moderate to heavy smokers.  Nevertheless, a relative  risk of lung cancer
of 38 would not be explained by differences in smoking  between the study
cohort and the Norwegian population.
    Davies  (1978, 1979) studied three chromate pigment  plants in England, of
which plants A and B produced both zinc and lead chromate, while plant C
produced only lead chromate.  The cohort of exposed  workers  consisted of
employees with _>. 1 year service that  were  first  hired  between 1933 and 1967
for plant A, 1948 through 1967 for plant B, and  1946 through  1961 for plant  C
(these years were governed by the availability of complete employment
records), and for which vital statistics were  available as of 1977.   Using
these guidelines, 396, 136, and 114 subjects were obtained from  plants A,  B,
and C, respectively, and these groups were further  subdivided into  high  and
medium exposure and low exposure groups.  The  observed mortality from lung
cancer in the different plants by exposure category  was compared to  the
                                       7-56

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the expected mortality as  calculated  from  national lung cancer mortality rates
for all  males in England and  Wales.   These data are presented 1n Table
The exposure categories of high  and medium were combined because they were
similar.  Adjustments to the  expected number  of lung  cancer death values were
made in the following manner: (1)  the expected number of lung cancer cases was
adjusted upward because the proportion of  unskilled and semiskilled workers in
the study population was  higher  than  in the general population and these
persons are known to smoke more  than  the national  average;  (2) the expected
number Was adjusted downward  for factory B and upward for factory C to  reflect
the respective differences in local  lung cancer mortality in comparison to the
national lung cancer mortality.   The  author did not state how these
adjustments were calculated.   An elevated  risk of  lung cancer was present only
in the high exposure groups in plants A and B, while  plant  C, which
manufactured only lead chromate  showed no  elevated risk.  Also, workers in
plant A that were hired after production modification in 1955 showed no
increased risk of lung cancer, even  though there was  a minimum follow-up
period of 15 years.  The  authors suggest that these data indicate that  zinc
chromate was associated with  the etiology  of lung  cancer, while lead chromate
was not, and although the  data was limited by the  small  sample size, the
authors claim that engineering controls had effectively lowered the  risk  of
lung cancer in plant A.
    Frentzel-Beyme (1983)  reported that the observed  number of lung  cancer
deaths exceeded the expected  among workers in five chronate pigment  plants  in
the Netherlands and West  Germany.   In only one  factory,  however, was this
excess statistically significant.   The authors  did not find a lung  cancer
mortality dose-response by intensity  of exposure or duration of exposure.   The
numbers of deaths in each  exposure category were  rather snail, however.
                                     7-57

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                     TABLE  7_12  •  LUNG CANCER IN WORKERS IN THE CHROMATE PIGMENT INDUSTRY
                                                  (navies 1979)
High and Medium Exposure
Plant and Year of Number of Observed Expected
Initial Employment Men Lung Cancer Lung Cancer
Plant A
1932 - 1954 175 18t 8.17
^ 1955 - 1967* 62 0 1.14
CO
Plant B
1948 - 1967 116 7§ 1.43
Plant C
1946 - 1967 95 1 2.46
Low Exposure
Number of Observed Expected
Men Lung Cancer Lung Cancer

77 2 2
14 0 0.16


20 0 0.1

19 1 0.37
    *Plant modification in Plant A in 1955 considerably reduced  employee  exposure  to  chromates.Thus,  results
were divided into two time periods, 1932-54 and 1955-67,  for  the purpose  of  analysis.
    tP < 0.01.
    §P < 0.001.

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Chromium Plating
    Royle (1975) studied  mortality in  the  chromium  plating  industry  of  England
in a retrospective study  between 1969  and  1972,  and also  reported  on the  first
2 years of a prospective  study which began in  1972.  Workers  in  this industry
are exposed to hexavalent chromium in  the  form of chromic acid mist  and some
sodium dichromate dust.   In the retrospective  study, 1,238 chrome  plating
workers employed for > 3  months were traced along with  1,284  manual  laborers
used as controls.  The control subjects were matched to the exposed  workers
for sex and the last date they were known  to be  alive,  while  the subgroup of
workers that were currently employed in the chrome  plating industry  were  also
matched for smoking habits.  Similar  success was achieved in  tracing both
groups, and from the response to questionnaires, there  was little  difference
in the smoking habits of  the groups.   The  death  rate was  higher  in the  chrome
plating group, with 109 deaths observed as compared to  85 in  the control
group.  In examining cause-specific deths, there was a  significant (P < 0.05)
difference in the death rate for cancer at all sites, 3.15% in chrome platers
as compared to 1.63% in controls, while deaths from malignancy of the lung and
from malignancy of the gastrointestinal tract  were  each increased  although not
significantly.  Other causes of death  were similar  in the two groups.  In a
small group (220 subj.ects) with high exposure, there was  greater mortality
associated with employment of 1 year  or more as  compared  with exposure  for
less than 1 year.  The results of the  first 2  years of  the prospective  study
indicate an increased proportionate cancer mortality with 12  cancer  deaths
(the tables indicate 12,  although the  text reports  9) of  33 deaths reported  in
the chrome-exposed group  compared with 3 cancer deaths  of 19  deaths  in  the
control group.
    In addition to the mortality study, Royle  also  conducted  a morbidity  study
                                     7-59

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of the chrome plating workers.  The controls were the same as those in the
mortality study.  The author stated that detailed results of the morbidity
study would be published in a separate article.  Among platers, it was
reported that there was a significant increase in the prevalence of a large
majority of different respiratory symptoms.  The possible causes of this were
examined in some detail.  In the course of this investigation, it was found
that a significantly larger proportion of controls (8.3%, 93 controls) than
platers (3.6%, 36 platers) had been engaged in asbestos processing.  Thus, the
risk of lung cancer due to chromium exposure in platers compared to that of
the controls may have been underestimated.  Results of this study are
inconclusive because of the relatively short (3 year) follow-up period in this
retrospective study, and because results from the prospective study are only
preliminary.
    Okubo and Tsuchiya (1979) conducted a cohort study of 889 Tokyo chrome
platers an unspecified number of controls selected from the same factories as
the chromium platers.  The follow-up period was April 1, 1970 to September 30,
1976.  Vital statistics were ascertained using the records of the Tokyo Health
Insurance Society of the Plating Industry.  The type of work and chromium
exposure history of the members of this society were investigated by a
questionnaire sent to the manager of each factory.  The recovery rate of the
questionnaire was 70.5%.  Survival  information on retired subjects was
obtained from the offices of the Japanese family registration system.  Among
the 889 male chromium platers, 19 deaths were observed or about 50% of that
expected.   The expected number of deaths was calculated using the annual  male
mortality rates, by age, for Tokyo.  In contrast, the authors reported a
slightly higher percentage of deaths in the control  group.
    The authors noted that two possible types of error may have occurred in
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this study:   1)  deaths from the chromium group  were  incorrectly  assigned  to
the control  group and 2) the 30% of nonresponders  may have  represented  a
significant  number of factories where chromium-related deaths  may  have
occurred.
    In addition  to the problems noted by the author, the follow-up period for
this study,  six  years, is probably not long enough to be able  to detect
differences  in lung cancer mortality resulting  from  chromium exposure.   Also,
the size of  the  cohort, 889, is relatively small  to  be able to detect
significant  differences in lung cancer mortality.
    Silverstein  et al.  (1981) found a statistically  significant  increase
(P < 0.001)  in the lung cancer proportionate mortality ratios  for  both  male
and female white employees in a die-casting and electroplating plant.  In this
plant workers were exposed to chromium during electroplating,  but  nickel  and
copper were also used to electroplate.  The other operations of the plant were
zinc alloy die-casting  and buffing, polishing,  and metal cleaning  of zinc and
steel parts.  Because of the employees' exposure to  other potential
carcinogens, no conclusion can be made from this study regarding the
association of chromium electroplating and lung cancer mortality.

Ferrochromium Workers
    Langard et al. (1980) studied ferrochromium workers at a ferrochromium
plant in Norway who were predominantly exposed to trivalent chromium.  In an
industrial hygiene study of the plant in 1975, ambient chromium levels  of
between  0.01 and 1.34 mg/m3 were detected in the ferrochromium department,
and, of  this chromium,  it was determined that 11 to 33% was hexavalent.  The
study consisted of 976  employees who worked for > 1  year, were alive after
1953, and initially employed prior to 1960.  The study group was divided into
                                    7-61

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10 subgroups by job description,  with  only  325  subjects  specifically
associated with ferrochromium  production  and  considered  exposed to  chromium.
Comparisons of cancer incidence  for  all  sites and  for  different sites  were
made between the Norwegian  male  population  and  ferrochromium  worker population
using the Norwegian Cancer  Registry.   Poisson distribution  was used to test
for statistical significance.   A comparison of  total mortality was  also made.
Of the 10 job-related subgroups,  only  the ferrochromium  workers had a
significant difference between the observed and expected number of  lung cancer
cases (7 observed versus 3.1 expected, P  <  0.05).   Note:  The statistical
significance was calculated by Langard et al. to be P  =  0.08. CAG  tested the
difference using the Poisson distribution and found the  significance to be
P < 0.05.  Total mortality  and cancer  incidence at all sites  were similar to
the general population.  The relative  risk  of lung cancer among the
ferrochromium workers as compared to the Norwegian male  population, may be
underestimated, however, since the county in which the plant  was  located,
Hordaland, had an age-adjusted lung cancer incidence 58% that of  Norway's  lung
cancer incidence rate.  If  this 58% is multiplied by the expected number of
lung cancer cases calculated from national  rates,  this newly  calculated
expected number is different from the  observed  at P <  0.01.  If nonchromium
exposed workers in this plant were used as a control  population,  the  risk  of
lung cancer in chromium-exposed workers is increased to  8.5,  which  is
significant at P < 0.01.  The ferrochromium workers were possibly exposed  to
two other carcinogens, asbestos and polycyclic  aromatic  hydrocarbons;  however,
this was unlikely to have been an important factor, the  author  indicated,
since a group of 243 ferrosilicon workers in the same  plants  were believed  to
be exposed to these two known carcinogens to the same  degree, and no  increased
risk of lung cancer (0 observed and 2.78 expected) among them was observed.
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However, the sample size of ferrosilicon workers (Number = 243) would have to
be considered to small to be able to detect any signifiant excess of lung
cancer.
     In studies of the ferrochromium industry in Sweden, Axelsson et al. (1980)
concluded that there was no association between employment in the
ferrochromium industry and risk of lung cancer.  The study cohort consisted of
1,876 men employed for one or more years between 1930 and 1975, all of whom
were alive as of 1951.  Observed cases were included for comparison if they
occurred 15 years after first exposure.  The exposed population was compared
with males in the county in which the plant was located, and comparisons were
tested for significance by two-sided P-values using the Poisson distribution.
The workers were subdivided into four groups consisting of arc-furnaces;
transport, metal grinders, sampling; maintenance; and office workers.  It was
estimated that these groups were exposed to 2.5, 0.5 to 2.5, 2.5, and 0
mg/m3 of Cr+3 and Cr°, and 0.25, 0.01 to 0.05, 0.05, and 0 mg/m3 of
Cr+6, respectively.  Medical  examination of employees during the last 3
years of the study detected three cases of perforated septum of the nose,
confirming that some exposure to Cr+6 had occurred in at least a portion of
the work force.  No statistically significant difference in cancer mortality
between that observed in the workers and that expected based on national
mortality data was found.   The only significant increase in cancer incidence
was an increase in respiratory cancer (4 observed and 1 expected, P = 0.038)
in the 315 maintenance workers.   Of the four cases of respiratory cancer, two
were diagnosed as mesotheliomas, and the authors suggested that these may have
resulted from exposure to  asbestos.  It was also noted that the control
population  was mainly rural  dwellers,  while the exposed group consisted  of
urban dwellers,  and that rural  residents in Sweden generally smoked less.
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From the results of this study, the authors concluded that  no association
existed between exposure to predominantly Cr"1"3 and Cr° and  the development
of cancer.  Because of the confounding due to smoking and exposure to
asbestos, no definite conclusions can be drawn from this  study.

Summary of Epidemiologic Studies
    Epidemiologic studies of chromate production  facilities in the United
States, Great Britain, West Germany, and Japan have all found an  association
between occupational  exposure to chromium and lung cancer.   Workers in  the
chromate production industry are exposed to both  Cr(III)  and Cr(VI) compounds.
Most of the epidemiologic studies did not attempt to determine which chromium
compounds were the etiological  agents.  Three studies of  the chrome pigment
industry, one in Norway, one in Great Britain, and the third in the
Netherlands and Germany, have also found an association between occupational
exposure to chromium and lung cancer.  The predominant chromium exposure in
the chrome pigment industry is  to Cr(VI).  One study of the chromium plating
industry in England (Royle 1975) reported that workers exposed primarily to
Cr(VI) (chromic acid mist and some dichromate dust) had a significantly
(P < 0.05) higher mortality from cancer at all sites than the control  group;
respiratory cancer mortality was not found to be  elevated,  however, and the
follow-up period was only for 3 years.  Thus the  results  are inconclusive.
Results of a Japanese study (Okubo and Tsuchiya 1979) of  chrome platers was
negative.  A proportionate mortality study by Silverstein et al.  (1981) of a
die-casting and electroplating  plant where chromium was used to electroplate
found an excess of lung cancer  mortality, but because of  the presence of other
carcinogens in the plant, no conclusions regarding the association of chromium
and lung cancer can be drawn from this particular study.  Two studies of the
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 ferrochromium industry reported an increased risk of lung cancer mortality.
     In most of the studies, smoking data was inadequate for any detailed
 analyses with regard to smoking as a confounding variable with respect to lung
 cancer.  However, the lung cancer relative risks found by most of the studies
 were higher than the risks that would be expected on the basis of difference
 in smoking habits between the study group and the controls.  The relative risk
 of lung cancer for smokers in the United States as compared to nonsmokers is
 about 10-fold (U.S. Department of Health and Human Sevices 1982).  In
 contrast, Brinton et al. (1952) found a relative risk of lung cancer mortality
 for chromate production workers of about 29; the odds ratio (unadjusted) of
 being a lung cancer case and being occupationally exposed to chromium was
 calculated by the CAG from the data by Baetjer et al. (1959) to be about 32
 for cases at the Johns Hopkins Hospital  and 23 for cases at the Baltimore City
 Hospital.  Langard and Norseth (1975) calculated a risk of dying from lung
 cancer for workers in the chrome pigment industry of 38.  Lastly, in regard to
 the issue of smoking as a confounding variable, the dose-response in terms of
 length of time worked in the industry seen in the Hayes et al.  (1979) and the
 Taylor (1966) studies and the dose-response found by Mancuso (1975)  in terms
 of mg/m3-years of total  chromium would not likely be explained  by
 differences in smoking habits between the study population and  the comparison
 population.  Using the International  Agency for Research on Cancer (IARC)
classification scheme for the assessment of human evidence of carcinogenicity,
the Carcinogen Assessment Group would classify the human evidence for chromium
as sufficient.
                                        7-65

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 QUANTITATIVE ESTIMATION



    This quantitative section deals with the unit risk for chromium in air and



 the potency of chromium relative to other carcinogens that the CAG has



 evaluated.  The unit risk estimate for an air pollutant is defined as the



 lifetime cancer risk occurring in a hypothetical  population in which all



 individuals are exposed continuously from birth throughout their lifetimes to



 a concentration of 1 ug/m3 of the agent in the air that they breathe.  This



 calculation is done to estimate in quantitative terms the impact of the agent



 as a carcinogen.  Unit risk estimates are used for two purposes:  1) to



 compare the carcinogenic potency of several  agents with each other, and 2) to



 give a crude indication of the population risk which might be associated  with



 exposure to these agents, if the actual exposures are known.



    The data used for the quantitative estimate can be of two types:  1)



 lifetime animal  studies, and 2) human studies where excess cancer risk has



 been associated with exposure to the agent.   It is assumed, unless evidence



 exists to the contrary, that if a carcinogenic response occurs at the dose



 levels used in a study, then responses will  also  occur at all  lower doses with



 an incidence determined by the extrapolation model.



    There is no solid scientific basis for any mathematical extrapolation



model  that relates carcinogen exposure to cancer  risks at the extremely low



 concentrations that must be dealt with in evaluating environmental  hazards.



 For practical  reasons such low levels of risk cannot be measured directly



 either by animal  experiments or by epidemiologic  studies.  We must,  therefore,



depend on our current understanding of the mechanisms of carcinogenesis for



guidance as to which risk model  to use.  At  the present time the dominant view



of the carcinogenic process involves  the concept  that most agents  that cause



cancer also cause  irreversible damage to DNA.   This  position is  reflected by
                                     7-66

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the fact that a very large proportion of agents that cause cancer are also



mutagenic.  There is reason to expect that the quanta!  type of biological



response, which is characteristic of mutagenesis, is associated with a linear



non-threshold dose-response relationship.  Indeed, there is substantial



evidence from mutagenesis studies with both ionizing radiation and a wide



variety of chemicals that this type of dose-response model  is the appropriate



one to use.  This is particularly true at the lower end of the dose-response



curve; at higher doses, there can be an upward curvature probably reflecting



the effects of multistage processes on the mutagenic response.  The linear



non-threshold dose-response relationship is also consistent with the



relatively few epidemiologic studies of cancer responses to specific agents



that contain enough information to make the evaluation  possible (e.g.,



radiation-induced leukemia, breast and thyroid cancer,  skin cancer induced by



arsenic in drinking water, liver cancer induced by aflatoxin in the diet).



There is also some evidence from animal experiments that is consistent with



the linear non-threshold model (e.g., liver tumors induced in mice by



2-acetylaminfluorene in the large scale EDf)j study at the National  Center



for Toxicological  Research and the initiation stage of  the two-stage



carcinogenesis model in rat liver and mouse skin).



    Because it has the best, albeit limited, scientific basis of any of  the



current mathematical extrapolation models, the linear non-threshold model  has



been adopted as the primary basis for risk extrapolation to low levels of  the



dose-response relationship.



    The quantitative aspect of the carcinogen risk assessment is included  here



because it may be  of use in the regulatory decision-making  process, e.g.,



setting regulatory priorities, evaluating the adequacy  of technology-based



controls, etc.   However, it should be recognized that the estimation of  cancer
                                     7-67

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 risks to humans at low levels of exposure is uncertain.  At best, the linear
 extrapolation model used here provides a rough, but plausible estimate of the
 upper-limit of risk; i.e., it is not likely that the true risk would be much
 more than the estimated risk, but it could very well be considerably lower.
 The risk estimates presented below should not be regarded as an accurate
 representation of the true cancer risks even when the exposures are accurately
 defined.  The estimates presented may be factored into regulatory decisions to
 the extent that the concept of upper risk limits is found to be useful.
    Although there are many epidemiologic studies demonstrating that chromium
 (Cr+6) is a potential human carcinogen, few of the studies provide adequate
 exposure data for use in risk estimation.  One study by Mancuso (1975)
 provides what the CAG feels is limited but adequate information for this
 purpose, however.  The Mancuso study was based on a cohort of 332 white male
 workers employed in a chromate plant between 1931 (when the plant began to
 operate) and 1937 and followed to 1974.
    In his study, Mancuso reported lung cancer death rates by levels of
 exposure to soluble, insoluble, and total chromium concentrations.  Because
 only lung cancer mortality for total  chromium exposure was reported by age
 group, the CAG has used the only dose-response data for total  chromium in its
 risk estimation.
    Exposure information in the Mancuso study was derived from an industrial
 hygiene study of the plant conducted in 1949.  In this study,  time-weighted
 averages of exposure to insoluble, soluble,  and total  chromium per cubic meter
were calculated for each occupation and for  each worker in every department.
Using this data and company personnel  records,  Mancuso was able to calculate
an estimate of exposure to soluble, insoluble,  and  total  chromium by duration
of exposure (in mg/m3-years)  for each  member of the 1931-37 cohort.   The
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plant began operation In 1931.  In 1949, after the industrial  hygiene study
had been conducted, the company initiated a comprehensive program designed to
reduce exposure to employees and improve manufacturing efficiency.  Until  that
time, however, the company had not undertaken any programs for the purpose of
reducing employee exposure.  It should be noted,  however, that Bourne and  Yee
(1950) who conducted the industrial  hygiene survey in 1949 reported that  "in
order to meet price and quality competition, improvements in  equipment and
processes have been made periodically during the  past 18 years, and it is  the
universal experience of industrial hygiene personnel  that greater process
efficiency is almost invariably associated with a more healthful  working
environment.  Therefore, there seems little doubt that atmospheric
contamination in the past was greater than in 1949."   Nonetheless, no
concerted effort was made to reduce employee exposure until  late in 1949  and
because this particular plant was a relatively modern plant  at the time of the
industrial hygiene survey, it is unlikely that improvements  in efficiency  over
the period 1931 to 1949 would have reduced employee exposure  to a great
extent.  Thus, the GAG considers Mancuso's data to be a reasonable
approximation of what workers in the study cohort were exposed to during  their
entire working history.  We recognize the possibility that the exposure may be
slightly underestimated because it seems likely that  a greater proportion  of
the "total exposure" was contributed by the exposure  prior to 1949 than post
1949.
    The effects of underestimating the exposure concentration, along with
other uncertainties on the estimation of potency, will  be addressed in the
Discussion section.
    As stated in the previous paragraphs, the CAG used only  Mancuso's data and
lung cancer deaths by total  chromium exposure in  its  risk estimation.  The
                                    7-69

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CAG, however, believes that only some of the hexdvalent chromium compounds can

for certain be said to be carcinogenic.  Thus, the risk estimate presented

here for total chromium will obviously underestimate the true risk from

exposure to those chromium compounds that are carcinogneic.   The risk estimate

and carcinogenic potency of chromium as discussed in the following pages

places chromium in the first quartile of those carcinogens for which CAG has

done risk estimates.  Thus, the potency of chromium compounds that are

carcinogenic is very high in comparison to that of other compounds for which

the CAG has done risk estimates.  This, the CAG feels,  is important

information, but it should be cautioned that it is to be used only in

conjunction with those chromium compounds evaluated by  the CAG to be

carcinogenic.




Data Available for Estimating the Cancer Risk

   Table  7-13  , which is taken directly from Mancuso  (1975), presents the

age-specific lung cancer deaths, the corresponding person-years, and the range

of exposure to total chromium.

    To estimate the lifetime cancer risk due to exposure to  chromium, we

assume that an exposure, D (mg/m3-years) , as presented  in Table 7-13   , is

equivalent to the continuous exposure d (ug/m3) calculated by



                   d = JL x __§. x 240 x 103 Ug/m3
                             24   365
where Le is the midrange in each age category,  f is the fraction of time in

age exposed, and 8/24 and 240/365 are the fractions of a day and year,

respectively, that a worker spent at the plant.   For instance,  if D = 8

mg/m3-years, Le = 60, and f = 0.65,  then d = 44.96 ug/m3.   The

assumption of f = 0.65 implies that  the cohort  exposure to chromium began
                                        7-70

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     TABLE  7-13  •  AGE-SPECIFIC LUNG CANCER DEATH AND THE GRADIENT EXPOSURE  TO TOTAL  CHROMIUM
                                           (Mancuso 1975)
Exposure to Total Chromium (mg/m^-year)
Age <1.00 1.0-1.99 2.0-3.99 4.0-5.99 6.0-6.99 7.0-7.99 8+*
45-54 Deaths 1 2
Person-years 886 459
55-64 Deaths 1 3
Person-years 707 356
65-74 Deaths 1 1
Person-years 235 166
24330
583 348 159 140 262
14231
462 250 113 98 203
21103
182 80 42 41 81
    *Data in the last column are not used in our risk assessment  because the range of exposure in
this class is not known and it does not  appear reasonable to assume  that all  three age groups had  an
identical exposure distribution in this  class.

-------
approximately at age 20.  The assumption is that the particular exposure
pattern (unknown to us) leading to the cancer mortality rates as observed is
equivalent to the continuous constant exposure starting from the age when
exposure began.  This assumption may or may not be realistic.  However, it
would be more unrealistic to make a different assumption concerning the
exposure pattern when all that is given is an exposure which itself was
calculated by taking the weighted average of the duration of exposure for each
respective job the man had.
    Since the person-year in each category presented in Table  7-13   is very
small, the exposure categories are combined as shown in Table  7-14   to
increase the statistical stability.  The last column of Table 7-14     is given
for the purpose of identifying which exposure categories in Table 7-13    are
combined.  The midrange of age and exposure concentration is used in
Table 7.^4   .  Data in this table are used to estimate the lifetime cancer
risk due to chromium exposure.

Choice of Dose-Response Model
    It has been widely recognized (e.g., Doll  1971)  that the age-specific
incidence curve tends to be linear on doubly logarithmic graphs, or
equivalently, the age-specific incidence follows the mathematical  form
where b and k are parameters that  may be related  to  other  factors  such  as
dose, and T may be one of the following  three  cases:
    1.  T is age when  cancer is  observed,
    2.  T is the time  from the first  exposure  to  observed  cancer,  or
                                        7-72

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                 TABLE  7-14   .   COMBINED  AGE-SPECIFIC  LUNG CANCER  DEATH  RATES
                            AND  TOTAL  CHROMIUM EXPOSURE (in ug/m3)
Age
50
50
50
60
60
60
70
70
Concentration
(ug/m3)* Death Person-year
5.66
25.27
46.83
4.68
20.79
39.08
4.41
21.29
3
6
6
4
5
5
2
4
1345
931
299
1063
712
211
401
345
Background Exposure Range
Ratet as Presented in Table 7-13
6.05 x lO-4
6.05 x ID'4
6.05 x lO-4
1.44 x IO-3
1.44 x IO-3
1.44 x ID'3
1.57 x IO-3
1.57 x IO-3
_< 1
2.0
6.0
<_ 1
2.0
6.0
1 1
2.0
.99
- 5.99
- 7.99
.99
- 5.99
- 7.99
.99
- 7.99
. r> ,
f = 0.65 in the formula described in the section  "Data  Available For Estimating  the  Cancer
Risk."  The concentrations presented in  this  table are  the averages  of  several exposure
categories weighted by corresponding person-years.
    tBackground rate is estimated from 1964 U.S.  Vital  Statistics.   The year 1964 is selected
because it is estimated that a large proportion  of lung cancer deaths occurred during that year.

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    3.  T is the time from exposure to cancer minus the minimum time for a
        cancer to be clinically recognized.


    This model has been shown to arise from the somatic mutation hypothesis of

carcinogenesis (Armitage and Doll  1954, Whittemore 1978, Whittemore and Keller

1978).  It has also been shown to arise from the epigenetic hypothesis when

the reversible cellular change is programmed to occur randomly (Watson 1977).

These authors and many others have used this model to interpret and/or

estimate potency from human data.

    Since the data that could be used for risk estimation are limited, a

simple model  that fits the data should be used.  Therefore, the observed

age-specific incidence is assumed to follow the model


                            I(t,d) = B(t) + h(t,d)


where B(t) is the background rate at age t and h(t,d) = Q(d) tk-1 with
                p
Q(d) = q.d + q d , a function of dose d.

    Once the parameters q^ q2, and k are estimated, the lifetime

cancer risk associated with an exposure d by age t, taking into account the

competing risk, can be calculated  by


                         t               s
                P(t,d) =/ h(s,d)exp{- [/ h(y,d)dy + A(s)]}ds
                         0               0


where exp[-A(s)] is the probability of surviving to age s and h(t,d) =

I(t,d) - B(t), the age-specific incidence after adjusting the background rate.



Estimation of the Risk Model
    To estimate the parameters  in h(t,d)  we assume,  as  is  usually  done,  that

the number of lung cancer deaths, X,  at age t,  follows  the Poisson
                                     7-74

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distribution with the expected value
                          E(x) = N x  (B + Q(d)  tk-!)

where N is the person-year associated with X, B is  the  background  rate  at  age
t, and Q(d) = q^ + q2d2.
    Using the BMDP computer program P3R and the theory  relating  the  maximum
likelihood and non-linear  least square estimation by  Jennrich  and  Moore
(1975), the parameters q1 , q?, and k  are estimated  by the  method of
maximum likelihood as q.  = 1.11 x 10~7, q2 = 1.84 x 10"9,  and  k  =
2.915.
    Thus, the age-specific cancer death incidence at  age t due to  chromium
exposure d ug/m3 is given  by
                            h(t,d)  = Q(d)  t

where
                 Q(d)  = 1.11 x 10'7d + 1.84 x lO'V

    The model  fits the data well  as one can see from the goodness  of  fit
statistic
                          X2 =z (0-E)2/E = 1.60

which has, asymptotically, a chi-square distribution with 5  degrees of  freedom
under the model  specified.  The observed and predicted  values  used in
calculating X2 are (3, 2.5), (6,  7.2), (6, 5.1), (4, 3.11),  (5,  6.7), (5,
4.1), (2, 1.4) and (4, 4.3).
    Taking into account the competing risk, the lifetime probability  of lung
cancer death due to exposure to chromium d ug/m3 is  given by
                                    7-75

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                    L
           P(L,d) = / h(t,d)exp{-[(Q(d)/2.915)  t2-915  +  A(t)]}dt
                    o
where L is the maximum human lifetime and is  mathematically  equivalent  to

infinity, since the probability of surviving  beyond  L  is  0.

     At low doses, approximately,


                            P(L,d) = d x P(L,1)


where P(L,1) is the lifetime cancer risk due  to  exposure  to  1  ug/m^ of

chromium.  The unit risk, P(L,1), has been adopted by  the CAG  as  an indicator

of the carcinogenic potency of a chemical compound.




Calculation of the Potency at 1 ug/m3

    To calculate the unit risk, P(L,1), we need  to know exp[-A(t)], the

probability of surviving to age t.  Since this probability can only be

estimated, it is assumed that the survival probability is constant over a

5-year interval as provided in the U.S. Vital Statistics.

    Using this approximation and by integrating  the formula, P(L,1), we have
P(L,1) = Z[exp(-3.87 x ICT* t^
                                       exp(-3.87 x 10-  t

       = i.ie x in-2



where (t^j, t-j) is a 5-year interval and PJ is the probability of

survival up to the age t-j.j.  PJ is assumed to be a constant over the

interval and is estimated from the 1975 U.S. Vital Statistics.
                                       7-76

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Relative Potency  of  Chromium
    Figure  7-1    1s  a  histogram  representing the frequency distribution of
potency indices  of 52  suspect  carcinogens  evaluated by the CAG.  Table 7-15
presents the potency index  for these  52 suspect carcinogens.  The potency
index for a compounds  is a  rounded-off potency expressed in term of
(mMol/kg/day)'1.   When human data  are available for an agent, they have been
used to calculate the  index.   When no human  data  are  available, animal oral
studies have been used in preference  to animal inhalation studies.
    Based on the occupational  study and the  assumption that the daily  air
intake of a 70 kg man  is 20 m3, the potency  index for chromium 1s calculated
as 4 x lb+3, which lies in  the first  quartile of  the  distribution.   This
provides the relative  potency  of chromium  in comparison  with  other suspect
carcinogens evaluated  by the  CAG.

JJI scussion
    The following discussions  are  intended to provide some  insight about  the
uncertainties on the estimated carcinogenic potency of  chromium.   No
statistical confidence limits  can  be meaningfully calculated  because it  is not
reasonable to assume that the  exposures were measured without error.
    1.  As noted previously,  the risk is estimated on the  basis  of the total
chromium obtained from all  the soluble and insoluble chromium to which workers
were exposed.  If indeed only  some compounds of  Cr+6 are carcinogenic, then
the potency presented  above is likely to be underestimated.  However, it is
difficult to evaluate  the level of underestimation because the proportion of
the total chro-nium exposure that is carcinogenic is unknown.   If one could
assume that the carcinogenic  chromium compounds  constituted half of  the total
chromium exposure for  each  of  the exposure groups in Table 7-13     , then the
                                      7-77

-------
                                  4th
                               quartile
   3rd
quartile
   2nd
quartile
   1st
quartile
                                      1x10
       4x10
       2x10
                            246
                         Log Of Potency Index
Figure 7-1    .   Histogram representing frequency distribution of the  potency
                indices of 52 suspect carcinogens evaluated by the Carcinogen
                Assessment Group.
                                   7-78

-------
TABLE  7-15.   RELATIVE CARCINOGENIC POTENCIES AMONG 53 CHEMICALS EVALUATED
  BY THE CARCINOGEN ASSESSMENT GROUP AS SUSPECT HUMAN CARCINOGENS1»2»3
Compounds
Acrylonitril e
Aflatoxin B}
Aldrin
Ally! Chloride
Arsenic
B[a]P
Benzene
Benzidine
"leryl 1 iun
Cadmium
Carbon Tetrachloride
Chlordane
Chlorinated Ethanes
1,2-dichl oroethane
hexachloroethane
1,1, 2, 2-tetrachl oroethane
1 ,1 ,1-trichl oroethane
1,1 ,2-t rich! oroethane
Chi oroform
Chromium
OOT
Dichlorobenzidi ne
1 ,1-dichl oroethylene
Oieldrin
Slope
(mg/kg/day)"1
0.24(W)
2924
11.4
1.19x10-2
15(H)
11.5
5.2xlO-2(w)
234(W)
4.86
6.65(W)
1. 30X10-1
1.61
6.90x10-2
1.42x10-2
0.20
1.6x10-3
5.73x10-2
7x10-2
41
8.42
1.69
1.47x10-1(1)
30.4
Molecular
Weight
53.1
312.3
369.4
76.5
149.8
252.3
78
184.2
9
112.4
153.8
409.8
98.9
236.7
167.9
133.4
133.4
119.4
104
354.5
253.1
97
380.9
Potency
Index
1x10+1
9xlO+5
4x10+3
9x10"!
2x10+3
3x10+3
4x10°
4x10+4
4x10+1
7x10+2
2X10+1
7x10+2
7xlOn
3x10°
3x10+1
2x10-1
RxlOn
8x10°
4x10+3
3x10+3
4x10+2
1x10+1
1x10+4
Order of
Magnitude
0°9io
Index)
+1
-t-6
+4
0
+3
+ 3
+ 1
+5
-t-2
-t-3
+1
+3
0
+1
-1
+ 1
-H
+4
+3
+3
+1
+4
                                                   (continued on the following page;
                                  7-79

-------
TABLE  7-15
(continued)
Compounds
Dinitrotoluene
Diphenylhydrazine
Epichlorohydrin
Bis(2-chloroethyl )ether
Bis(chloromethyl ) ether
Ethylene Dibromide (EDB)
Ethylene Oxide
Formaldehyde
Heptachlor
Hexachlorobenzene
Hexachl orobutadi ene
Hexachl orocycl ohexane
technical grade
alpha isomer
beta isomer
gamma isomer
Nickel
Nitrosamines
Dimethyl nitrosamine
Diethylnitrosamine
Di butyl nitrosamine
N-ni trosopyrrol i di ne
N-nitroso-N-ethylurea
N-nitroso-N-methyl urea
N-nitroso-di phenyl ami ne
PCBs
Slope
(mg/kg/day)"1
0.31
0.77
9.9xlO-3
1.14
9300(1)
8.51
0.63(1)
2.14xlO-2(I)
3.37
1.67
7.75xlO-2

4.75
11.12
1.84
1.33
1.15(W)

25.9(not by q*)
43. 5 (not by q*)
5.43 l
2.13
32.9
302.6
4.92xlO-3
4.34
Molecular
Weight
182
180
92.5
143
115
187.9
44.0
30
373.3
284.4
261

290.9
290.9
290.9
290.9
58.7

74.1
102.1
158.2
100.2
117.1
103.1
198
324
Potency
Index
6x10+1
lxlO+2
9X10-1
2x1 0+2
lxlO+6
2x1 0+3
3xlO+1
6X10-1
lxlO+3
5xlO+2
2X10+1

lxlO+3
3xlO+3
5xlO+2
4x10+2
7xlO+1

2xlO+3
4x1 0+3
9x10+2
2x10+2
4xlO+3
3x1 0+4
1x10°
lxlO+3
Order of
Magnitude
(1o9lQ
Index)
• +2
+2
0
+2
+6
+3
+1
0
+3
+3
+1

+3
+3
+ 3
+3
+2

+ 3
+4
+3
+2
+4
+4
0
+3
                           (continued  on  tne  following  page)
           7-80

-------
                                 TABLE 7-15
(continued)

Compounds
Phenols
2,4,6-trichlorophenol
Tetrachlorodloxin
Tetrachloroethylene
Toxaphene
Trichloroethylene
Vinyl Chloride

Slope
(mg/kg/day)'1
1.99xlO-2
4.25xl05
5.31X10'2
1.13
1.26xlO-2
1.75x10-2(1)

Molecul ar
Weight
197.4
322
165.8
414
131.4
62.5

Potency
Index
4x10°
lxlO+8
9x10°
5xlO+2
2xlOn
1x10°
Order of
Magnitude
0°9lp
Index)
+1
+8
+1
+3
0
0
Remarks:
    1.  Animal  slopes are 95% upper-limit slopes based  on  the linear multistage  model.
        They are calculated based on animal  oral studies,  except  for those  indicated  by  I
        (animal  inhalation), W (human occupational  exposure), and H  (human  drinking water
        exposure).  Human slopes are point estimate,  based on linear non-threshold model.

    2.  The potency index is a rounded-off slope in (mMol/kg/day)"-^  and  is  calculated by
        multiplying the slopes in (mg/kg/day)"^  by  the  molecular  weight  of  the compound.

    3.  Not all  the carcinogenic potencies presented  in this  table represent  the same degree
        of certainty.  All  are subject to change as new evidence  becomes available.
                                           7-81

-------
risk at low doses  would  be  approximately twice as much as that estimated by
our model.
    2.  As  indicated  previously, there  1s  a possibility that the use of 1949
hygiene data may result  in  a  slight  underestimation of what workers were
actually exposed to.   However,  because  the plant was  relatively modern at that
time, the underestimation is  unlikely to be very considerable.  If an
underestimation of 2  tines  were assumed, then the unit risk would be reduced
from 1.2 x  10~2/ug/m3 to 6.0  x  lCT3/ug/m3.
    3.  The risk presented  in this  report  may be somewhat overestimated in the
sense that  it is implicitly assumed  that the smoking  habits of chromate
workers were similar  to  the general  white  male population, while it is
generally accepted that  the proportion  of  smokers is  higher for industrial
workers (thus the higher background  incidence rates)  than for the general
population.  As a  sample calculation, we found that if the background rate of
lung cancer mortality for the cohort in Table  7-14     is  increased by 40%,
then the corresponding unit risk would  be  reduced by  about 25%, or from 1.2  x
10-2 to 8.7 x 10-3.
    The background age-specific rate of lung  cancer at ages 50, 60, and 70
could be 40% more than those  presented  in  Table   7-14   , should it be  assumed
that 80% of c-iromate  workers  are ever-smokers  (individuals who  smoke  at least
100 cigarettes during their lifetimes)  and only  50% of the  general white  male
population are ever-smokers.   It could  also  result  from  other assumptions
about the smoking habit of  the  chromate workers  and the  general population.
For instance, the background  rate could be increased  by  approximately 40%,
based on the age-specific  cancer rates  provided  by  Doll  (1971), if the
proportion of smokers in each smoking level  is distributed  as  follows:
                                    7-82

-------
                   NUMBER OF CIGARETTES SMOKED PER DAY
                          0    1-14    15-24    25 or more
Chromate Workers         0.3     0.2     0.3        0.2





General  Population       0.5     0.2     0.2        0.1
                                   7-83

-------
SUMMARY



    It is pressumed that all  forms of Cr(VI)  are carcinogenic but the degree



of carcinogenicity is modified by the solubility of the specific compounds.



Using the International  Agency for Research on Cancer (IARC)  criteria, animal



studies have provided sufficient evidence for the carcinogenicity of the



following Cr(VI) compounds:  calcium chromate, strontium chromate, and zinc



chromate.  Both Cr(III)  and Cr(VI) compounds  have been ineffective in



producing lung tumors by inhalation in animals .  Similarly,  negative results



have been obtained following  the ingestion of Cr(III).  Chromium has been



shown to be carcinogenic by intrabronchial, intrapleural,  intramuscular



implantation, or subcutaneous injection;  however.  Cr(III)  compounds have  been



studied less extensively than Cr(VI); however, animal  studies indicate that



Cr(VI) is more likely to be the etiologic agent in human chromium-related



cancer.



     The epidemiologic studies of chromium have demonstrated  an  association



with respiratory cancer  in chromate-producing industries.   The strength of the



association is evidenced by the high relative risks of lung cancer and the



consistency of results by different investigators in different countries.



Results of three epidemiologic studies of chrome pigment workers are also



suggestive of an association  with lung cancer.  Less clear, however, is



question of which form of chromium is carcinogenic.  One epidemiologic study



of chrome pigment workers (Davies 1978, 1979) suggested that  zinc chromate was



carcinogenic while lead  chromate was not.  However, the data  on  the lead



chromate pigment workers was  limited by small  sample size.   Most of the



epidemiologic studies did not attempt to  distinguish the carcinogenic species



of chromium.



    Although a number of epidemiologic studies have found  an  association
                                       7-84

-------
between exposure to chromium and lung cancer, the data that could be used for
estimating the cancer risk due to the exposure to chromium are limited to the
study by Mancuso (1975).  Mancuso (1975) reports age-specific lung cancer
mortality data for chromate production workers in terms of total  elemental
chromium exposure.  Using this information, CAG estimated the lifetime cancer
risk due to a constant exposure to air containing 1 ug/m^ of elemental
chromium to be 1.2 x 10"^.  This is considered an upper-bound estimate,
since it is based on a model  that is linear at low doses.

CONCLUSIONS
    Using the IARC criteria,  epidemiologic studies provide sufficient evidence
that chromium is a human carcinogen.  Using IARC criteria, the animal  bioassay
studies have provided sufficient evidence for the carcinogenicity of
hexavalent chromium.  The carcinogenic evidence for trivalent chromium  is
inconclusive.  Hexavalent chromium is mutagenic in multiple tests while the
data for trivalent  chromium is inconclusive.
    Using the IARC classification scheme, the level  of carcinogenic  evidence
available for the combined animal  and human data would place  chromium into
Group 1, meaning that there is decisive evidence for the  human carcinogenicity
of chromium.
                                      7-85

-------
7.2.2.    Genotoxicity.








     7.2.2.1.  JJJ VITRO MJTAGEMCITY — In an attempt to understand the funda-



mental  biological activity  of  metals and its  relationship  to carcd.nogenesis,



numerous in vitro experiments have been conducted.  Many of these studies attempt



to exploit the strong relationships between molecular events involved in muta-



genesis and carcinogenesis.   In particular, the interaction of xenobiotics with



nucleic acids  is  believed  to be a  critical  event in mutagenesis  and/or cell



transformation.  Cultures  of mammalian  cells  and bacteria, as well as cell-free



systems, have been used to explore the potential mutageni city/carcino geni city of



chromiun salts.  Although mutageni city assays  employing bacterial  tester strains



have received widespread use in screening compounds for mutageni city (e.g., Ames



assay), these  assay systems have  not  been frequently  employed  for  screening



metals. Nevertheless, positive results  have been obtained with several metals in



both in vitro mutageni city assays and assays using DNA repair deficient bacteria.



A summary of the results obtained from  in  vitro mutagenicity assays of chromium



compounds are presented in Table 7-17.



     In an early study of the mutagenicity of chromium in bacteria, Venitt and



Levy (1971*) tested three soluble Cr(VI) salts (Na2Cr04, KgCrO^, and CaCrO^) for



mutagenic activity using a spot test.  Following application of each compound to



the plate at levels  of  0.05,  0.10,  or 0.20 jimol, a 3-fold increase was observed



in the number of  reversion colonies in the tester organism E.  coli WP2 (try").




Similar results were observed with  both  E.  coli  WP2 and WP2 uvrA  (absence  of



excision-repair) were exposed to Na^CrOj.  in suspension, followed  by removal  of



the test compound and plating on minimal  plates.   Repair deficient  E.  coli WP2



(exrA,  error prone DNA  repair) and WP2 uvrA were compared with wild type E. coli



WP2 for mutagenic response  in  the spot  test  to KpCrOj..   Since  all  tester
                                     7-86

-------
                     TABLE 7-17



The In Vitro Mutagenloity Bloassay of Chronic Compounds

Test
Reverse mutation E.



Reverse mutation E.
E.
Reverse mutation E.
E.
E.
Reverse mutation E.
E.
^j E.
i
GO
~~-j Reverse mutation E.
Reverse mutation E.

Reverse mutation E.
E.
Reverse mutation S.
Indicator
Orgmiam
coll W2 (try")



coll W>2 (try")
coll W2 uvrA
coll M>2 (try")
coll V*>2 uvrA
coll H?2 exrA
coll H>2
coll H>2 uvrA
coll CM571

coll Hs30R
coll Vi>2

coll K>2
coll B/r WP2
typhimiriia
Conpoir.d
Tested
Na_CrO,

K CrO,
I*— \>i\y ,
CSCrO,J
Na CrO,
Na,Crt)!
£ H
K CrO,
fCCrO?
2 H
2 "
KpCr.O-
JC Cr 0
K^Cr^oI
I— £ *
K 2Cr^
K2C"2°7
K CrO
2 1»
K2Cr20?

ohr ornate*
Valence
State
46

•*6
46
46
46
46
46
46
46
46
46

46
46
46

46
46
46
Metabolic
Activation
no

no
no
no
no
no
no
no
no
no
no

no
no
no

no
no
no
Dose
0.05, 0.10, and
0.20 [imol/plate


0.5 to 5 mg/nfc
0. 5 to 5 mg/ttt.
0. 05 fimol/plate
0. 05 nmol/plate
0. 05 n mol/plate
1. 0 x 10~^M
1.2 x 10 ,M
1.2 x lO'^M

13. 0 x 10"^M
13.0 x TO'^M
0. 5 |ig/nA

NR
NR
NR
TA92


Reverse mutation S.


typhlmurlun
diohr ornate*

chr ornate*
46

46
no

no
TA1 978. TA92












di chr ornate*

chr ornate*

diohr ornate*

46

46

46

no

yes

yes

NR

100 and 200
nmol/plate
100 and 200
nmol/plate
100 and 200
nmol/plate
100 and 200
nmol/plate
Application Response
spot test 4-

4-
4-
suspension assay +
suspension assay +•
spot test +
spot test 4
spot test 4-
suspension assay +•
suspension assay +
suspension assay

suspension assay 4-*
suspension assay +*
fluctuation test 4-

spot teat 4-
spot test
plate 4-
incorporation
plate +
incorporation
plate 4
incorporation
plate +
incorporation
plate
Incorporation
plate
incorporation
Refer en oe
Venitt and
Levy , 1 971


Venitt and
Levy , 1 971
Venitt and
Levy , 1 971

Nisid oka, 1975



Nakamtro et al .,
1978
Green et al . ,
1976
K an em at s u
et al., 1980
L of roth and
Ames, 1978


Lofroth, 1978







-------
TABLE 7-17 (oont .)
Indicator Compound
Test Or^nisn Tested
Reverse mutaticn S. typhlmurium K,Cr?07
TA1535, TA1537,
TA1538, TA100,
TA98
Reverse mutaticn S. typhlmuriun Na2Cr20_
Na_Cr_0,
2 27
Reverse mutaticn S. typhlmuriun Na.Cr-O-
TA1535, TA1537
TA98, TA100 CrO,
3
-~j CaCrO..
i ^
co K_Cr,(L
d. 2 f
2nCtQh'7t\W)y
«l e.
Na_Cr 0_
~2 2-y
Crt).
3
CaCrO,,
H
1C Cr,0_
2 2 Y
aiCrOa«3h(OH)_

Forward mutaticn S ohl aos acoharomyces K?Cr_0_
pom be
Forward mutaticn Chinese Hamster KpCr.O.
to 8-aaguanine V79 cells ZnCrO^
resistance FbCrO,.
Valence
State
46


46
46

46
46

46

46

46

46

46

46

46

46

46

46
46
46
Metabolic
Activation Dose
no


no
yes

no
no

no

no

no

yes

yes

yes

yes

yes

no

no
no
no



10 to
10 to

10 to
10 to

10 to

10 to

10 to

10 to

10 to

10 to

10 to

10 to

102nM

NR


10 ng/plate
80 ng/plate

200 ng/ pi ate
200 (ig/plate

200 tig/plate

200 (i ft/ pi ate

200 (ig/plate

200 |ift/ pi ate

200 us/ plate

200 ng/plate

200 lift/ pi ate

200 |ig/plate



0.35 to 0.78 lie/at.
1 to
5 to
4 Hft/nA
10 Hft/nA
Application Response
spot test


plate 4-
incorporation
plate
incorporation
plate 4-
incorporation
pi at e 4^
incorporation
plate 4-
incorporation
plate +
incorporation
plate 4-
incorporation
plate
Incorporation
plate
Incorporation
plate
incorporation
plate
incorporation
plate
incorporation
suspension 4-'

in culture medim +
in culture medim +•
In culture medium
Reference
Kanematsu
et al . , 1 980

DeFlora, 1978;
Petrilll and
DeFlora, 1978

Petrel 11 and
DeFlora, 1977,
1978

















Bonatti et al .,
1976
New bo Id et al .,
1979


-------
                                                                      TABLE 7-17 (cont.)
 I
GO
Test
Gene conversion
Gene conversion
Reverse mutation
Reverse mutation
Reverse mutation
Reverse mutation
Indicator
Orgmism
S ohi aos accharom ycea
pom be
Saccharemyces
cerevisiae D7
E. coll (try") Cr
E. coll Ft30R
y
S. typhimuriun
TA98, TA1537,
TA1535, TA100
S. t yphimuri un
TA100
Compound
Tested
K2Cr207
Cr03
•2( 33 a^SO,, -210,0
Cr(CH3COO)-
CrtCCSO^.-^H-O
CrCl-«6fLO
CrK(SO,t)2.12H20
Valence
State
46
+6
+3
+3
+3
+3
+3
+3
+3
Metabolic
Activation Dose
no 102 to 105nM
no 10~2 to 10~3
no NR
no 130 x 10~3M
no/yes 20 mg/plate
no/ yes
no/ yes 800 ^g/ pi ate
no
no
Application Response
suspension +
suspension +
spot test*
suspension assay +•
plate
incorporation
plate
incorporation
plate
incorporation
Reference
Bonatti et al .,
1976
Fukunajp et al .,
1982
Venitt and
Levy, 1971
Natemiro et al .,
1978
Petrilli and
DeFlora, 1977
Petrilli and
DeFlora, 1978a,b
    • = Refer to text  for fvrtner information.

    NR =  Not reported

-------
organisms  yielded approximately the sane mutagenic response regardless of the



presence or absence  of DMA  repair pathways, the authors concluded that  chromium



directly interacted with  DNA,  with subsequent mispairing  of  bases occurring



during cell division.  Other possible explanations for these  results  were not



considered.   The soluble  Cr(III)   compound ,  Cr^SO^SO^ «2H20  [This  was the



formula  given  in  the   report, although   the   test   compound   was   probably



Cr?(SOi|)-?»K_SOi. «24H20.],   gave   negative  results  (specifics  in  experimental



protocol were  not  given), along with soluble salts of tungsten, molybdenum,  zinc,




cadmiun, and mercury.



     Nishioka  (1975) obtained positive mutagenic  results  with  K2Cr_07(VI) in  a



suspension assay  using the tester strain E. coli WP2 and WP2 uvrA, but not in the



tester strain E.   coli CM,-™, .   Since  strain CMC_,  is  a re combination- deficient
              —  -   571
strain, it was postulated that metal mutagenesis needed seme  component  of  the



recA allele.  Positive results were also reported for K  Cr 0  by Kanematsu et  al .



(1980) in the spot test  using strain  E.  coli  WP2; however,  E.  coli B/r  WP2  was



negative  (no  dose information  was presented  in this  study).   Nakamuro  et  al .



(1978) used E. coli HsSOR,  a  uvrA  minus mutant requiring arginine , to study  the



mutagenicity of K2Cr2CL(VI) and KjCrO^VI) ,  and Cr(CH3COO)3( Ill) in a suspension



assay.  The  respective mutagenic frequency of these compounds was 13^»  ^5, and 30


            Q

mutants x 10  viable cells.   Although positive mutagenic responses were reported



for all test substances, the low survival of between  iJ and 10$ at the doses which



were reported as  mutagenic may lead  to  artifacts,  particularly if  spontaneous



revertants arise during growth on the trace levels of arginine used to supplement



the top agar, as suggested  by Green  (1976).



     Green et al .  (1976)  also observed mutation in E.  coli WP2  using a fluctua-



tion test.  At  levels  of 0.5 (ig/md K^CrO^,  '\H8 of  250 tubes were positive as



compared to 64 in controls.  At a high dose of 2.5 ng/m2,f  only 10 of 150 tubes
                                     7-90

-------
were  positive  as compared with 31* tubes in  the  controls.   The apparent  anti-



mutagenic effect at the  higher dose was attributed to the toxicity of  the K?CrOj..



      The mutagenicity of chromium compounds has been assessed in the Ames  assay,



which uses  specially constructed  histidine dependent  strains  of  Salmonella



typhijaurium; however, in  the  original report on the validation of  this  assay,



McCann  et  al.    (1975) stated  that  the test was not  suitable for metals as  a



result  of the  high  concentration  of magnesium, citrate,  and  phosphate  salts in



the media.   Chromate  and  dichromate  but  not  the  chromic ion (the  specific salt



was not mentioned) were reported in an abstract to cause frameshift mutations in



,3. typhimurium strains containing the his D3052 and his C3076 mutations, as well



as his G 46 strains containing the R factor.  The greatest response was  observed



in TA92 with  five  re vert ants/nmol  Cr.  Kanematsu  et  al. (1980)  has  reported



negative results with  K^Cr^Q^ in the spot  test using strains TA1535, TA100, TA98,



TA1537, and  TA1538; however,  the lack of  exposure  information in this  report



makes it difficult to  evaluate.  In a recent  study of the mutagenic  activity of



chromic chromate (18.5$ Cr(VI),  and 50$ total chromium) in S_.  typhimurium  strain



TA1535, Witmer et al. (1982)  reported that  mutagenic  activity,  expressed  as



number  of revertants - on  test plates -f number  of  revertants  on  spontaneous



plates, was increased  as the standard salt concentration of the minimal media was



reduced.  At least  a  portion of  this  observed increased  activity resulted from



the poor survival of the cells in the low  salt media  and the  associated  decrease



in the  number  of  spontaneous  colonies.   Although  the  author  suggested that



variation in  the j>.  typhimurium assay may  be useful in further evaluation of the



mutagenic activity of  inorganic compounds such as chromiun, further  validation



of this method  will be required before these results can be compared with the



other  available data on  mutagenicity of chromiun.
                                     7-91

-------
     In further studies,  Lofroth  (1978) found that chromate and dichromate were




mutagenic in strain TA1978 as  well  as TA92.   On addition of a mammalian micro-




somal activation system (S-9 mix) prepared from the livers of Aroclor pretreated




male rats, the number of revertants decreased from  206 and  357 (assays without




activation system) to 89 and 89 (assays with activation system) for TA1978 and




TA92, respectively.  In the absence of NADP in the S-9  mix,  the inhibition of




chromium  mutagenicity  did not occur.   It was also reported  that  addition of




Cr(VI)  to an aqueous solution of  NADP  did  not change  the  valence  state of




chromium.  It was  concluded that NADP plus additional factors in the  S-9 mix were




necessary to  inactivate Cr(VI), possibly  by  reduction to Cr(III).   Similarly,




Petrilli  and  DeFlora (1977)  obtained positive  responses with Na^Cr^d-,  CrO-,




CaCrCL, and K Cr-CL when assayed  in strains TA1537,  TA1535,  TA98, and TA100 at




levels  between  10 and 200 |ig/plate.  When the response was expressed as rever-




tants/jig  of  Cr(VI) ,  there was no  statistically  significant  difference in the




activity of each of these compounds.  The Cr(III) compounds,  CrK(SO.)2 and CrCl_,




gave negative results.



     DeFlora  (1978)  and  Petrilli  and DeFlora  (1978a,b) further  studied the




effect  of metabolic activation on the mutagenicity of Cr(VI) and (III) .  DeFlora




(1978)  observed  a decrease in the mutagenicity  of Na?Cr_CL  in S_.  typhimurium




strain  TA100 in the presence  of S-9 mix prepared from  rat liver.  When Na_Cr20™




was added at 40 ng/plate,  the number of revertant colonies decreased from 705 to




M20, 370, 283, 228, and 221 with  the incorporation, respectively, of 0, 10, 20,




30, 40, and  50 \ii of S-9  fraction/pi ate.  Petrilli and DeFlora  (1978b) extend




these   observations   to   the   Cr(VI)   compounds   NapCapO-,   CrCL,   KpCrO^,




ZnCrOj»2i(OH)p (zinc yellow also contained 10$ CrO_), and PbCrCL'PbO, with muta-




genic activity in strains TA1535,  TA1537, TA98, and TA100  detected in the absence




of S-9 mix but not in the presence of S-9 mix, prepared  from rat liver.   Little or
                                     7-92

-------
no loss of rautagenic activity occurred when the added S-9 was prepared from lung



or muscle.  Also, human plasma added to the top agar had no effect on chromium



mutagenicity,  but lysates of erythrocytes  did result in  the  loss  of mutagenic



activity.  The loss of mutagenicity was associated with reduction potential of



components in the S-9  mix,  with  reduced GSH,  TPNH, and  G6PD plus S-9  mix (to



generate TPNH)  all  inhibiting the mutagenicity of Cr(VI) .  In a  study using



varying levels of TPNH, the mutagenic response was  correlated with the amount of



Na_Cr?07 (originally 52 |ig Cr(VI)Xplate) that remained as Cr(VI) .   Conversely,
Cr(III) as CrK(S04)2, CrCl3, and CKNO^, which were inactive in the Ames assay,



were  converted  to active mutagens  by  the  addition of  nontoxic  levels of the



strong oxidizer  KMnOj.  ( Petrilli and DeFlora,  1978a).    The presence  of  this



oxidizing  agent  reversed the  effect  of S-9  mix on the mutagenicity of Cr(VI) .



These data supported the  conclusions that the valence state of chromium was an



important  factor in producing a mutagenic response in the Ames assay, and that



naturally  occurring biological reducing agents were capable of reducing Cr(VI)



to Cr(III) .  Although the studies  show  that  Cr(VI) compounds produce a positive



response,  these studies do not indicate whether Cr( III)  or (VI) is the ultimate



mutagen which interacts with the DNA of the  cell.



     Cr(VI) compounds have also been demonstrated to  be mutagenic  in vitro in



eucaryotic test  systems, while Cr(III)  was inactive.   Bonatti  et al . (1976)


                                                2        5
exposed  yeast,  Schizosaccharomyces  pom be ,   to  10   to 10 \M  of KpCr-O-  in  a



forward mutation  assay and a test  for gene conversion.  Forward mutation was



observed in 7 of 480,054 colonies  exposed to 10  (iM K^r^Oj for 7 hours, which



was not statistically increased from controls in which no mutants were observed



in 84,546 colonies; however, if comparison was made to the historical spontaneous



mutation   rate  (no  mutants   in  10   colonies),  then  chromium  significantly



increased the mutation rate.   In the study  of gene  conversion, there was a dose
                                     7-93

-------
related increase in the four allelic combinations  examined.   The authors noted



that there were limitations in this study, but that the data suggest that K Cr 0



waa mutagenic under the test  conditions.



     Using another  strain of yeast, Sac char omyces cerivisiae U?,  Fukanaga et al.



(1982) examined the genetic activity  of  chromium  trioxide.   The  compound was


                                                 -2       9
incubated with  the  cells  at  concentrations  of 10   to  10   M for a  period of



24 hours. Following incubation,  the  cells were plated to  determine viability and



recombinance with cross-over in the ade locus.   The highest concentration tested



caused nearly 100$ cell death while viability  at the lowest  concentration was



11%.   The cross-over  frequency in  treated  cells  was dose dependent  with the


  _o

10~  M concentration resulting in  a  frequency  of  1.4$ as compared  to  0.02$ for



untreated cells.  It was also reported that  only growing and not resting cells



were susceptible to the effects of chromium,  suggesting  to the authors  that



chromiixn may affect the fidelity of DNA polymerase.



     Forward mutagenesis to 8-azaguanine resistance in cultured Chinese hamster



V79/1* cells was used by Newbold  et al.  (1979) to assess the genetic activity of



both Cr(VI) and (III).   The highly soluble  and slightly soluble Cr(VI) salts,



K?Cr_07  and 2iCrOj,, respectively,  both produced   dose  related increases  in



8-azaguanine resistant  cell   colonies, while  the  insoluble Cr(VI)  compound,



PbCrOj., and the soluble Cr(III)  compound, Cr(CH_COO)_, were inactive.   The more



soluble KpCr_0_ was =5-fold more active than the  ZnCrCL.   It  was stressed that



both the valence state  and solubility of a chromium compound affected  its muta-



genic  potential.    Although   high  solubility  was  necessary for  a  mutagenic



response, the authors suggested  that low solubility would facilitate a carcino-



genic response, since  compounds  of  lower solubility might remain in  the  body



longer, providing a low dose continuous exposure to chromium.

-------
     7.2.2.2.   EFFECTS  CN DMA AND  DNA  REPLICATION  — It is  apparent  from  the



results  of  bioassays in  both prokaryotic  and eucaryotic  systems  that sane



chromiun compounds are mutagenic.   In general, soluble Cr(VI)  compounds were



positive in reverse and forward mutagenicity assays,  while  insoluble salts  and



Cr(III) compounds  are inactive.  Metabolic activation  inhibited the mutagenicity



of Cr(VI),  probably  by  the  reduction of  the Cr(VI)  to  Cr( III) by  cellular



reducing agents such  as GSH.   Other studies have been performed which indicate



that chromiun  compounds  interact  with DNA  and decrease the fidelity of  DNA



polymerase, and this  mechanism may  participate in chromium induced mutagenesis.



     The difference in the  sensitivity of recombination deficient  strains  of




bacteria as compared  to wild  type to the toxic effects of chromium compounds  has



been used as an indicator of DNA damage.  Nishioka (1975) exposed both rec~  and



rec+ strains of Bacillus  subtil is to K2Cr207(VI), K^rO^CVI), and CrCl^III)  by



placing 0.05 n& of 0.05 M solution  on  a filter  disc placed on the agar surface.



For the two Cr(VI) compounds, the zone  of growth inhibition was greater for  the



rec~ strains as compared with the rec+, indicating DNA  damage,  while there was no



difference in the  size of the zone of inhibition in  the assay  of CrCl..   The



positive rec-effect with  K^Cr 0  was diminished sharply when the reducing agent,



Na?SO,, was mixed with the test compound, indicating that the Cr(VI) oxidation



state was necessary to cause  DNA damage.  Similarly, Kanematsu et al.  (1980), in



a survey of 127 metal compounds in the  E. subtil is rec assay,  reported that three



Cr(VI) compounds  (KpCrCL, K^Cr 0„, and CrCL) were  positive at  0.05 md of a 0.005



and 0.1 M solution, respectively, for  the salts and oxide.  Cr(III),  Cr^SO^,,



was negative, and it  was  noted that the sample was toxic to the bacteria at  the



dose tested.



     Nakamuro et al. (1978)  examined  three Cr(VI)  compounds (K2Cr20>, K^rO^,



and CrO_) and three Cr(III)  compounds  (Cr(CHLCOOK, Cr(NO_)_, and CrCl-) in  the
                                     7-95

-------
rec~ assay with J3. subtills.  The compounds were tested  using 0.02 md aliquots of




solutions between 3.2 x  10~1  to  1.6x  10"  M for the Cr(VI) compound and 1.3 to




1.6 x 10"  M for  theCr(III) compound.  All compounds tested were positive except




for CrCl (III).   Although DNA damage as indicated by the  rec effect was observed




with   two   of   the  Cr(III)   compounds,  the   order   of   activities   was




K2Cr2°7 > K2Cr°4  > Cr°3  > Cr(CH3°°)3 > Cr(NO_)_, with the Cr(VI) compounds being



more active than  the Cr( III)  compounds.




     In an attempt to determine the mechanism of DNA damage induced by chromium,




Levis et al. (1978) examined the effects of KpCrpO™  exposure on DNA synthesis in




cultured BHK cells.  The cells  were  treated with 10~3,  10"**,  10~5,  and 10"6 M




K_Cr 0  for 1 to  4 hours, followed by  determination  of 3H-thymidine uptake into




DNA  and  the intracellular  pools.   At  the  two intermediate  doses,  increased




specific activity was observed in both the nucleotide pool and  DNA,  which was




attributed to chromium effects on the membrane transport of  the exogenous labeled




thymidine.   When DNA synthesis  was adjusted for the  changes in  the specific




activity of the pools, transient  and reversible inhibitions  of DNA synthesis were



              -6                                                      -5
observed at 10    M, and ccmplete  irreversible inhibition occurred  at 10   M.,  It




was reported that inhibition of DNA synthesis occurred prior to, and to a greater




extent than, inhibition  of  protein or  RNA  synthesis.  Although these  effects on




DNA synthesis may be partly related to the effects  of chromiun on the cellular




membrane, Raffetto (1977) reported evidence of  direct interaction with DNA in




A18&jR  cultured  cells,  as  indicated by  unscheduled DNA  synthesis  following




treatment with K-CrJD7 (VI).   In cells treated for  1 hour with  16,  4,  or  1 \ig




Cr(III)/m2, (as CrCl-), no increase was produced.  This  evidence suggests that at




least Cr(VI) can  alter DNA synthesis by more than a  single mechanism.




     Loeb and coworkers  (Sirova  and Loeb, 1976; Loeb et al.,  1977) have demon-




strated that chromiun compounds  adversely  affect the fidelity of DNA transcrip-
                                     7-96

-------
tion.  The compounds, CrCl- and CrOg, were incubated with avian myeloblastosis


virus DNA polymerase, a template with restricted base composition, and comple-

                                          •32
raentary deoxynucleoside radiolabeled  with J  P and noncomplementary deoxynucleo-


side labeled with 3H.  Following incubation,  the error frequency was  determined



to be greater when either metal compound was added to the incubation system as



compared to the controls.  The error  prone avian myeloblastosis virus DNA poly-


merase lacks exodeoxynuclease active  for removal of  noncomplementary  base, thus



providing optimum conditions for detecting the nonfidelity of DNA  synthesis.  In



later studies, Tkeshelashvili et al.  (1980)  demonstrated a similar increase in


misincorporation of complementary bases  using synthetic polynucleotides  and CrO_



and CrCl  in an assay system containing E. coli DNA  polymerase.   Both compounds



appeared to  have  approximately the same  activity.   Nearest neighbor  analysis


indicated  that the  noncomplementary  bases  were  incorporated  as  single base


substitution.   Noncomplementary nucleotides were also incorporated with native



DNA  from bacteriophage  $x174  (am3) and  E. coli DNA polymerase in vitro in the



presence of CrO_.   Infidelity in DNA  synthesis was detected  by loss of  an amber


mutation which was assessed by growth of infected strains of E.  coli  nonpermis-



sive and permissive for this mutation.


     Tamino  et  al.  (1981)  examined  the  in  vivo and in vitro  binding of both


Cr(VI) (K_Cr207) and  (III)  (CrCl_) to  DNA from  cultured BHK cell, and i.n  vitro to


commercial calf thymus DNA  and synthetic  polynucleotides.   The interactions of


Cr(III) with these nucleic acids iji vitro was dependent  on the G  and  C  content,


with synthetic poly(G)  and poly(C) binding  1.05 chromium ions per 100 nucleo-


tides.  Iri vitro and in vivo reaction also changed the ultraviolet  (UV) absorp-


tion spectra and thermal stability of the DNA.  When  Cr(VI) was used there was no



observed change in the  UV spectra whether exposure occurred i_n vitro or  i_n vivo.


The manner of treatment with Cr(VI) did  affect the thermal stability  of the DNA
                                     7-97

-------
with in  vitro exposure resulting in  the same types  of changes observed  with




Cr(III), while little change in thermal  stability occurred with in vivo exposure




to Cr(VI).  The data indicate that Cr(III) has some capacity to cross biological




membranes  and  interact with cellular DNA,  and that the types  of  interactions




between chromium and polynucleotides may be different for Cr(VI) and (III).




     Although  in  vitro  mutagenicity  indicated  that  Cr(VI)  compounds  were




generally active and Cr(III) compounds nonactive, the assays of chromium induced




DNA damage  in the rec  assay,  unscheduled DNA synthesis in mammalian cultured




cells, and increased jLn vitro infidelity of DNA synthesis indicated that at least




some activity is associated  with both valence states  (Nishioka, 1975; Nakamuro et




al., 1978; Raffeto, 1977; Loeb et al., 1977; and Tkeshelashvili  et  al.,  1980).








     7.2.2.3.  CHROMIUM    INDUCED    CHROMOSOMAL    ABERRATIONS    AND    CELL




TRANSFORMATIONS — Iri vitro, chromium  has  been shown to result  in the appearance




of chromosomal aberrations and  cell  transformation (Table 7-18).  Fradkin et al.




(1975) observed morphologic  changes  and loss of anchorage dependent  growth in




BHK21  cells treated with  0.25  and 0.5 ng  of CaCrO^»2HpO.   Chromate transformed




cells maintain the ability to grow independent of anchorage even after the cells




were re-isolated and  freed  of exposure to CaCrO^»2ELO.  Tsuda  and Kato (1977)




observed morphologic transformation  in primary hamster embryo  cells after expo-




sure to K2Cr20,,(VI)  at a level of 0.1,  0.2, and 0.5 \ig/wSi  for  24 hours.   The




transformation frequency  increased  from  0.179$ for  the  control  cultures  to




0.760,  2.86,  and  2.70? for cells exposed to 0.1,  0.2, and 0.5 |ig/m!l,  respec-




tively.    A  transformation frequency  of  2.10$ was  achieved with  the  positive




control, N-methyl-N'-  ni tro-N-ni tros oguan i dine (MNNG) at a level of 0.5 \ig/mH.




Conflicting results  of exposure to  CrCl-(III) and  K?Cr?0 (VI)  on  morphologic




transformation of BALB/c mouse  fetus cells in culture was obtained in two experi-
                                     7-98

-------
                        TABLE 7-18



Chromium Produced Clastogenlc Effects and Cell Transformation
Test
Cell transformation



Cell transf orm at i en




Cell transf crmaticn

Cell transf crmaticn

vj> Host mediated oell
V-D transf crmaticn

Clastogenic

Clastogenic







Clastogenic


Clastogenlc

Clastogenlc


Indicator
Cells
BHK21



primary embryo
hamster cells



BALB/o cells

primary embryo
hamster cells
primary embryo
hamster oells

BALB/c oells

CHO cells







mouse FM3A cells


hamster embryo
cells
V79 cells


Compound
Tested
CaCrO^O



K Cr20_
C- C. f



CrCl
K2Cr2°7
CaCrO,
K Cr04
ZnCrOjj
NaCrCv


CrCl
K2Cr207
K2Cr2°l
Na Cr267
K.CrCv
Na CrOu
CrO
CaC?Cv
CrClJ*
Cr(NO )
KCr(SOJ.
CKCOOOC)
K2?r2°7
CrO,
Cr2?S04)3
K2Cr2°7
C- C. \
K2Cr207


Valence
State
46



46




4-3
46
46
46
46
46


+3
46
46
46
46
46
46
43
+3
43
43
46
46
+3
46

46


Dose
0.25 and 0. 5 ug/mt



0.0, 0.1, 0.2, and
0.5 |ig/n£



0.04 and 0.4 tig/at
0. 015 and 0. 1 |ig/mt
0.006 mM
0.01 mM
0. 01mM
2.5 iig/100g body
weight on day 11
of gestation
0.04 and 0.4 ng/n*.
0.015 and 0.1 tig/mt
0. 1 and 0.3 ng/mt
0. 1 and 0.3 ng/nA
0.25 |ig/mfc
0.25 ng/mt
0. 1 and 0.25 \ig/JA
0.5 jig/ml
5 and 50 ng/nA
50 and 150 ng/mt
150 ng/nfc
5 and 20 |ig/n£
3.2x10^ to 3.2x10"^M
i.oxio^: to I.OXIO~;M
3.2x10 to 1.0x10~3M
0.5 iig/mi

0.35 to 0.8 |ig/nA


End point
Morphologic altera-
tions , loss of
anchorage dependent
growth
Morphologic altera-
tions



Morphologic altera-
tions
Increased suscep-
tibility to viral
tranf crmatlcn
Morphologic altera-
ations

Chromosomal aberra-
tions
Chromosomal aberra-
tions/ 100 cells
(17 in control cul-
tures )





Chromosomal aberra-
tions

Chromosomal aberra-
tions
Chromosomal aberra-
tions

Response
+



Transf crmatlcn
frequency increased
frcm 0. 76% for con-
trols to 2. 70< for
the high dose group
+«
+•
2 x control levels
2 x control levels
4 x control levels
+


4 at 0. 4 ug/mt
4- at both doses
36 and 56
58 and 169
42
41 and 65
60 and 62
102
21 and 31
1 5 and 21
32
38 and 39
4
4

+.

4


Reference
Fradldn
et al.,
1975

Tsuda and
Kato, 1977



Raffetto,
1977
Caato et
al., 1979
DiPaolo
and Casto,
1979
Raffetto,
1977
Levls and
Majone,
1979






Umeda and
Nishimura,
1979
Tsuda and
Kato, 1977
New bold
et al .,
1979

-------
                                                                   TAB1£ 7-18 (cont.)
Test
Clastogenic




Clastogenic


Clastogenic







^,
i
— Clastogenic
o
o



Clastogenic

Clastogenic


Claatoganic



Clastogsnie

Claatogenic

Indicator
Cells
cul tired hum an
leukocytes




cultured human
lymphocytes

CHO cells









Don Chinese
hamster cells



cultured human
lymphocytes

polychromatic
erythrocytes from
NMRI mi 08
gel cells from
Boleophthalmus
dussunierl

human lymphocytes

human lymphocytes

Compound
Tested
K2Cr2°7
K2C™4
Cr(CH-COO),

Cr(NO )
CrCl J J
K Cr 0
CrCl* '

K Cr20-
Na_Cr20,
K-CrOj.
Na-Crfj,
cro
CrCl
Cr(NOJ
ff* M / On. V
ftX/ I \ Owj. ) A
CrfCOOCHp

CrCl «6H,0
Cr 2(*0~'
4fc,0
Ci°L
it— CrOi,
K2Cr207
CaCrCv
CrO,
K CrO,
<_ *T

Na Cr 0



CrO

Cr03

Valence
State
+6
+3

+3
+3
+6
+6

+6
+6
+6
+6
+6
+3
+3
4-3
4-3

4-3
4-3
46
46
46
46
46
46


46



46

46

Dose
0.125 to i». OxID^M
0.5 to 8. OXIOT'M
U to 32 x 10 M
£
32.0 x 10"7M
32.0 x 10 M
10 , to 10~?M
10"° to lO^M

0. 1 and 0.3 )ig/mt
0. 1 and 0. 3 (Jg/nA
0. 25 ng/nft
0.25 ng/n*
0.1 and 0.25 ng/nft
5 and 50 ug/nft
50 and 150 (ig/nA
150 ng/nfc
5 and 20 ng/nA

32 ug/nA
0.32 [ig/nA
0. 8 }ig/nA
0. 8 |ig/BA
0. 025 to 0. 1 (ig/nA
0.01 to 0.02 (ig/nA
0.025 to 0.1 jig/nA
2 x 48.5 fflg/kg, IP
2 x 24.25 mg/ kg, IP
2 x 12.12 mg/kg, IP
1 or 5 mg/kg IM

24 or 30. 5 ppm in
the aquaria! water
Occupational exposure

Occupational exposure

End point Response
Chromosomal abarra- +
tions +
4- ,
at >16 x 10 M
„
-
Chromosomal abbera- +
tions

Sister chromatid 4-
exchange 4
+
+
+
—
M
_
-

Sister chromatid +•
exchange
+,
+
4-
Sister chromatid +
ex: hangs +
*
Micron ucleus test 4-
+
-
Chromosomal aberra- +.
tions

+
Sister chromatid 4-
ewhanga
Chromosomal aberra- +•
tions
Reference
Nakamuro
et al .,
1978



Stella
et al.,
1982
Levls and
Majone,
1979







Ohno et al . ,
1982



Gomez-Arroyo
et al., 1981

Wild, 1978


Krishnaja
and Rege ,


Stella et al .
1982
Sarto et al .,
1982
IP =  I ntra peritoneal  injection
IM =  Intramuscular  injection

-------
ments by Raffetto (1977).  Although both experiments showed an increase  in  the




number of transformed foci, this increase was not dose-related, and there was  a




10-fold difference between  the  two experiments performed under similar  condi-




tions.  The authors suggest  caution in interpreting these findings.  Casto et  al.




(1979), using  a different  experimental  system, demonstrated  that the  Cr(VI)




compounds,  CaCrOy, KpCr.,  and  ZnCrO^,  at concentrations  of  0.006, 0.01,  and



0.01 mM enhanced the  virally induced transformation frequency  of  primary embryo




hamster cells.   The  increase  in transformations  was  =2 for the Ca and  K salts,




and  4  for the  Zn salt.  Other  metals  assayed that had  no indication  from  the




literature of genotoxic properties did not enhance the rate of viral  transforma-




tion.



     Increase  in cell  transformation  has also occurred  in cells  isolated from




hamster embryos exposed in utero to Cr(VI) (DiPaolo  and Casto, 1979).  Initially,




it  was demonstrated that   NaCr01»t»fLO  at levels  of  1.0,  2.5,  and  5.0 jig/mi




increased  the  transformation frequency  of  isolated hamster  embryo   cells  in




culture by 0.7,  2.1, and  3.W%,  respectively.  Following this  initial  study,




pregnant Syrian golden hamsters  (number not reported) were given intraperitoneal




injections  of NaCrO^ at  2.5 mg/100 g body weight on day 11 of gestation.  It was




reported that cells isolated from embryos excised 2 days  later produced elevated




numbers of transformed  colonies; however, the data were not presented for these




studies.   It was concluded that transplacental exposure to chrcmate resulted in




alterations of fetal cells.



     A number of studies have also demonstrated that exposure of cells in culture




to  either  Cr(VI) or  (III) compounds produces chromosonal aberrations.   Raffetto




(1977) exposed BALB/c mouse cells to CrCl-dll)  and K2Cr207(VI) for either 48 or




96 hours.   Increase in the number of chromosomal  aberrations was noted for Cr(VI)




at  a level of  0.1 ug/md, but not for Cr(III) at the same level after a U8 hour
                                     7-101

-------
exposure;  however, with 96 hours of exposure, Cr(III) at 0. U ng/mJ,  (O.OU




was also tested) and Cr(VI) at 0.1 and 0.015 (ig/mX, produced  significant  (PcO.05)




increases in the number of chromosomal  aberrations.   Newbold et  al.  (1979)  also




detected dose dependent chromosomal damage with K Cr_0? at  levels between  0.35




and 0.8 ng/m?, in V79 cells.  Similar results were reported  by Levis and  Majone




(1979),  in which  both  Cr(III)  compounds  [CrClg,   Cr(N03)3>  KCr(SO)||)2,  and




Cr(COOCHj_] and Cr(VI) compounds (K2Cr2CL, Na2Cr207, K^rO^, Na^rO^, CrO_, and




CaCrCL) produced increases in chromosomal aberrations in CHO cells  in  culture.




Again, Cr(VI) compounds were more  active, with positive responses obtained  at




doses  of  0.1  to 0.5 ng/mJl, while marginal  positive responses  with Cr(III)




occurred at  doses  of  5 to  150  |ig/mH.   Although  both  valence  states  produced




chromosomal  aberrations,  only  Cr(VI)  induced  increases   in  sister chromatid




exchange, and even the increase  observed  with Cr(VI) was small in  comparison  to




the response elicited  by  the positive  control, Mitomycin C.  Sister-chromatid




exchange was also observed by Ohno et  al. (1982) in Don Chinese hamster cells




following treatment with the Cr(VI) compounds CrO ,  K^CrQ^,  and  K2Cr2CL at  con-




centrations of  0.32,  0.8,  and  0.8  fig/mS,, respectively.   The Cr(III) compounds




CrCl •6H?0 and  Cr_( SCLK'UfkO  were less  active  than the  Cr(VI)  compounds  at



levels of 32 and 6 [ig/m&, respectively,  with the increases due to the  sulfate not




significantly elevated over control values.  Again, as in the previous study, the



levels of chromium induced sister-chrcmatid exchange was 10 times less than  that




of  the positive control,  mitomycin C.   However, in  the  study  by Umeda and




Nishimura (1979), only the Cr(VI) compounds,  K2Cr2
-------
     The effectiveness of  chromium  compounds  in producing chromosomal aberra-




tions was affected by the  valence  state  of the  compound.  Tsuda and Kato (1977)




demonstrated that 0.5 ng/mSl of K Cr-CL produced chromosomal aberrations in 51 %




of hamster embryo metaphases examined;  however,  addition  of the reducing agent,




Na SO ,  at a level of 0.645 |ig/mS,  resulted in a decrease  in abnormal metaphases




to 6%.  This provides further evidence that  Cr(VI) was more active than Cr( III)




in producing chromosomal  aberrations.



     Nakamuro et al .  (1978)  used peripheral blood human leukocytes in short-term




culture to  elevate the clastogenic  effects  of  a number  of  Cr(VI) and Cr(III)




salts.  Following isolation and 24 hours of culture, the cells were exposed  to




Cr(VI)  (K2Cr20?, K2Cr04)  and Cr(III)  (Cr(CH3COC»3, CKNO^, CrCl^  chromium  at




the  respective  concentrations  of 0.125  to 4,  0.5 to  8,  4  to  32,  32, and




32 x 10"6 M.  Significant  increases  (P<0.01)  in total chromosomal aberrations
were observed at  the  higher doses with all compounds  except  CrtNO^)^ III)  and




CrCl (III).  It was apparent that human leukocytes responded similarly to other




cultured cells in regard to sensitivity to chromium induced chromosomal aberra-




tions.



     In vivo  clastogenic activity of  KCrO^ has  been  observed  in mice  by  the




micronucleus test (Wild, 1978).  NMRI mice (4 animals of each sex) were treated




by two intraperitoneal injections of  0.0,  12.12, 24,  25, or 48.5 mg/kg of J^CrO^




in saline, followed in  6 hours by examination  of  bone  marrow for micronucleus-




containing erythrocytes .  Of the 1000 cells examined per mouse,  there were 3.1,




4.8, 9.6, and 15.0/K micronucleated polychromatic erythrocytes in the control and




treated animals,  respectively.  The responses  in the  two  highest  dose groups




represent significant increases over control values.  Chromosomal aberrations,




including  breaks,  fragments,  rings,  exchanges and  unclassified makers,  were




observed in the gills  of the fish Boleophthalmus dussumieri  after  exposure to
                                     7-103

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 sodiun dichronate(VI) (Kushnaja and Rege, 1982).  The number of total aberrations




 increased  from  1 to  6 and  16 after intramuscular injection of Na Cr~0  at doses




 of 0.0, 1, and 5 rag Cr/kg, respectively.  Increases  in total aberrations of 0, 7,




 and 7 were also observed  when the fish were maintained in  water  for 96 hours




 containing chromium  at  levels  of 0.0, 24.0,  or  30.5 ppm (the high  dose level




 killed  50% of  the fish).    Other  heavy metals  and mitomycin C also produced



 chromosomal aberrations in this test system.




      Human lymphocytes  exposed either  in  vitro to  chromium or isolated  from




 workers  occupationally exposed  to  chromium  have been  used  to  investigate




 chromium induced chromosomal damage.   Increase in both sister chromatid exchange




 and chromosomal aberrations were observed when cultured hunan  lymphocytes  were




 exposed to the  Cr(VI) compounds K^r^, CaCrO^, and CrO_  (Gomez-Arroyo, 1981;




 Stella et  al., 1982); however,  the  Cr(III) compound, CrCl , was totally inactive




 (Stella et al., 1982) in this assay.   In blood samples taken from workers  in the




 chromium plating industry  (Stella  et  al., 1982; Sarto et al.,  1982)  there  were




 increases  in  chromosomal changes in the  peripheral  lymphocytes.  Stella  et al.




 (1982) obtained blood samples  from 12 male workers and  10 control  donors  not




 exposed to chromium.   There was no significant difference in  the  incidence  of




 sister chrcmatid exchange in the 5 older workers (24 to 47 years of  age);  how-




 ever, in the  7 youngest workers (17 to 23 years  of  age)  there was a significant




 increase.  The control subjects had  an  age associated increase in sister chroma-




 tid exchange,  while the  level in the exposed  population appeared constant.  Sarto




 et  al. (1982)  studied the peripheral lymphocytes of workers in two "hard" plating




 (one of the plants  was the  same as  that studied  by  Stella et  al., 1982) and two




 "bright" plating industries.   In the  "hard" plating industry,  exposure was  to




chromium alone, while  in  the  "bright" plating  industry the workers were  also




exposed to  nickel.  As in the previous study,  the younger  workers had significant
                                    7-104

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increases in  chromosomal  aberrations while the  number  of aberrations was  not




significantly elevated from control levels in  the older workers.  There was also



a positive association between  urinary chromium levels and the number of chromo-



somal aberrations in the two "hard" plating plants.  The authors concluded that




these data support the genotoxicity of the soluble Cr(VI) ion.



     7.2.2.4.  SUMMARY — In vitro and in  vivo assays of genotoxicity have tried



to  clarify  the mechanism  of chromium carcinogenicity  and have  supported  the



potential for Cr(VI)  to  be  the active carcinogenic species.   Cr(VI) has demon-




strated  consistently  postive  mutagenic  activity  in  a number  of  bacterial



systems.  This activity was demonstrated in the absence of a metabolic activation



system.  In the presence of a mammalian activation system,  the mutagenic activity



of  Cr(VI)  disappeared.   Because Cr(VI)  shows only marginal,  if any, mutagenic



activity, it was  suggested that the mammalian enzymes or  cofactors  in the activa-



tion system reduced Cr(VI) to Cr(III).  Both Cr(III) and Cr(VI) have been demon-



strated to interact with DNA in  bacterial assays,  and Cr(VI)  has inhibited DNA




synthesis and increased unscheduled DNA synthesis in mammalian cells in culture.



In in vitro studies,  both Cr(III) and Cr(VI) have  increased the infidelity of DNA



replication.  As  observed  with interaction with  DNA, both valences of chromium



have  been  demonstrated to produce clastogenic effects  in mammalian cells with




Cr(VI) being more active than Cr(III).  The effects observed included a variety



of  chromosomal  aberrations, sister  chromatid exchange, and the  appearance of



micronuclei  in  polychromatic erythrocytes.   Increased  chromosomal damage also



has  been observed in  human lymphocytes  cultured  from  subjects  occupationally



exposed  to chromium.   For  all the  observed genotoxic  effects, it  has  been



suggested that Cr(III) may  be the predominant intracellular species a result of




the reduction of absorbed Cr(VI) by  cellular  components.
                                     7-105

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 7.2.3.    Developmental Toxicity and Other Reproductive Effects.








      7.2.3.1.  DEVELOPMENTAL  TOXICITY  ~ Chromium salts  have been shown to  be



 teratogenic and embryotoxic in mice and hamsters following intravenous or intra-




 peritoneal  injection.   Golden hamsters  were  given single intravenous CrO  (VI)




 injections  of 5,  7.5,  10,  or 15 mg/kg  on  the eighth day of gestation  (Gale,




 1978), or an intravenous Cr03 dose  of 8 mg/kg  on day 7, 8,  9,  10, or 11 (Gale and



 Bunch, 1979).



      In  the first  study using mice,  treatment groups were as follows:   15  dams



 exposed  to 5 mg/kg,  18  dams  exposed to 7.5  mg/kg,  21  dams exposed to  10 mg/kg,




 and  H dams  exposed to 15 mg/kg.   In all but  the 5 mg/kg group, dams  exhibited



 signs  of chromium  toxicity,  including  weight loss and tubular  necrosis of the



 kidneys.  The highest dose was lethal  to 75% of  the dams.



     Chromium given  on  the eighth  day  of gestation resulted in increased fetal



 wastage  at  doses  of >7.5 mg/kg (statistics not  reported).  The major external




 anomaly  noted was  cleft palate.   Incidence  of  cleft palate  was significantly



 elevated above the control level  for all  treated groups (method not stated).  The



 primary  internal   abnormality recorded  was  hydrocephaly, with  an  increased



 incidence noted in all treatment groups.  In addition, a wide range of skeletal



 defects were noted.



     In the second study,  six pregnant  female hamsters were treated in  each time-



 group  with  three  concurrent  controls.  Fetal  death was greatest in litters



 treated  on  the seventh  day  of gestation  when  there was an 8H%  incidence of



 resorptions.  Resorption incidence was elevated to a small  degree on days 8 and 9



 (20 and 13$, respectively), but was not affected on days  10 and 11.




     As in  the previous  study,  cleft palate was the primary external abnormality



recorded, occurring  most  frequently in  litters  from  dams exposed on  day 7 of



gestation and  at elevated  incidence rates with respect to controls on days 8 and






                                    7-106

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9, but not on days 10 and 11  (determined based on tables  of binomial  confidence




limits).   The  only  internal  abnormalities  noted were kidney defects  in  groups




exposed on days 7 or 8 of gestation.  Group sizes were too small  to  establish  a




significant difference from controls.  As in the previous study, treated females




exhibited signs of chromium toxicity which included body weight loss  and tubular




necrosis of the kidneys.



     In a  continuation  of this work, Gale (1982) tested Cr(III) in  noninbred




(LVG) and inbred (CB, LHC, SDH, MHA, and PD4)  strains of hamsters for  reproduc-




tive and  teratological  effects.  The dams (10 animals/group) were treated by




intravenous injection on the  8th day of  gestation with a single dose  of 8 mg/kg




of  Cr(III).   On  termination  at day  15 of gestation,  it was determined  that




strains CB, LHC, and PD4 were resistant to any adverse effects  of  treatment.  In




the other  strains,  LVG, LSH,  and MHA  there was  an increased incidence of cleft




palate (33, 18, and   15 animals, respectively, while  control groups had no  more




than 1 pup/group with cleft palate)  and  other  external abnormalities.   The  dams




of  the  strains  with the  highest  incidences  of  fetal abnormalities  also  lost




weight during the period  of gestation following  treatment.  Strain  differences




were reported  for the reproductive effects  of other metal  compound in these




animals; however, the genetic  mechanism which results in these  differences in




susceptability were  not  understood.




     Matsumoto  et al.  (1976)  administered chromium  chloride  (+3),  19.52 mg/kg




(as Cr) , intraperitoneally to ICR mice  on  day  7,  8,  or  9 of gestation.  Eight




control dams received a.  saline injection.  Treatment group sizes were 5, 6, and  7




on  days 7,  8,  and 9, respectively.   The embryonic  and  fetal  death rates  were




elevated following treatment  on days 8 and  9  (P<0.001, method  not stated).   The




incidence of external malformations was significantly greater  than in controls




in litters of dams  exposed on day 8 of  gestation (PcO.001,  method not stated).
                                     7-107

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Recorded anomalies included exencephaly and open eyelids.  In addition, there was
a small increase in skeletal  defects in litters from dams treated on days 8 and
9.
     In a second experiment,  pregnant dams were given intraperitoneal injections
of chromium chloride at dose levels of 9.76,  14.64,  19.52, or 24.4 mg/kg as Cr on
day 8 of gestation. Group sizes were:  control (11 dams), 9.76 mg/kg (13 dams),
14.64 mg/kg (13 dams),  19.52  mg/kg  (7 dams), 24.4 mg/kg (9 dams).  Fetal weights
were significantly reduced in all  treatment groups (P<0.05,  method not stated).
The incidence of  external malformations was significantly greater  than control
values for the 14.64 mg/kg dose group and all higher dose groups (P<0.05, method
not stated).   Anomalies  with  greatest incidence  in rats  were open  eyelids,
exencephaly, and  acephalia.   In addition, skeletal  defects  were noted in these
same groups.
     lijima et al. (1979) dosed pregnant ICR mice with CrO (VI) by subcutaneous
injection.  Mice were injected  with a single 10 or 20 mg/kg dose on day 7, 8, 9»
10, or 11 of gestation.  The  20 mg/kg  dose was lethal to one-third of the dams.
In this  group,  a significant  increase  in external  anomalies was  recorded in
litters from dams exposed on day 8  of  gestation.   Cleft palate was the primary
abnormality.  In addition, increased fetal and embryonic death rates were noted
in groups exposed to  20 mg/kg on day 8  or day 11.
     The four studies  discussed in the  text above are summarized in Table 7-19.
     As  discussed previously  (Section 5.1.4.),  existing  data indicate  that
inorganic Cr(VI) and (III) are  not transported across the placenta to any appre-
ciable extent during  the period of organogenesis.   These data  indicate that the
observed teratogenic  effects  may well  be the secondary result  of maternal toxi-
city.
                                     7-108

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                 TABLE 7-19



Teratogenic and Fetotoxic Effects of Chromium
Compound Route Species
CrO, i.v. hamster
CrO, i ,v . hamster
i
o
CrCl, i .p . mouse
CrO- i.v. hamsters
3 (strain
LVG)
hamsters
(strain
CB)
Dose
5, 7.5, 10,
or 15 mg/kg
on day 8 of
gestation
8 mg/kg on
day 7, 8, 9,
10, or 11 of
gestation
9.76, 14.64,
19.52, or 24.4
mg/kg on day
8 of gestation
8 mg/kg on day
8 of gestation
8 mg/kg on day
8 of gestation
Fetal Effects
increased fetal death in
7.5, 10, and 15 mg/kg
groups, increased incidence
of cleft palate in all
groups, hydrooephaly and
skeletal defects
increased fetal death
following administration
on day 7, increased
incidence of cleft palate
following administration on
days 7, 8, or 9
depression of fetal weights
in all Cr treated groups,
increase in rate of external
abnormalities for groups
treated with 14.64, 19.52,
or 24.4 mg/kg
increased incidence of cleft
palate
no effect
Maternal Effects Reference
depressed weight gain and Gale, 1978
kidney tubular necrosis
at all doses above 5 mg/kg
weight less, tubular necrosis Gale and Bunch, 1979
of kidneys
not reported Matsunoto et al., 1976
body weight loss Gale, 1982
no effect

-------
             TABI£ 7-19 (oont.)



Teratogenio and Fetotoxlo Effects of Chromium
Conpound Route Species
hamsters
(strain
LHC)
hamsters
(strain
--J LSH)
i
— hamsters
0 (strain
PD4)
hamsters
(strain
MHA)
CrO. s.o. mouse
Dose
8 mg/kg on day
8 of gestation
8 mg/kg on day
8 of gestation
8 mg/kg on day
8 of gestation
8 mg/kg on day
8 of gestation
10 or 20 sag/kg
on day 7, 8, 9,
10, or 11 of
gestation
Fetal Effects Maternal Effects Reference
no effect no effect
increased incidence of cleft body weight loss
palate
no effect no effect
increased incidence of cleft body weight loss
palate
increase in external malfcr- lethal to 1/3 of dams lijiaa et alt, 1979
mations in 20 mg/kg group
when dosed on day 8, as well
as Increase fetal death when
dosed on day 8 or 11

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     7.2.3.2.   OTHER REPRODUCTIVE EFFECTS — Behari et al.  (1978) examined the



effects of  chromium  on testicular tissue  in  rabbits.   Animals  were injected



intraperitoneally with  chromium nitrate  or potassium dichromate  at doses of



2 mg/kg for 3  or  6 weeks.   Both  forms  of chromium resulted  in  a decrease in



testicular  succinic  dehydrogenase  at  3  and 6 weeks.   Adenosine  triphosphatase



was inhibited at both time  points with both compounds, but the Cr(III) compound



resulted  in  more  severe  depression.    Acid  phosphatase  was   significantly



depressed at 6 weeks in anjjnals given  the trivalent salt.



     The administration of  Cr(III)  resulted in thickening of the tunia albuginea



and congestion of blood  vessels  following 3 weeks of treatment.   In addition,



cells in the seminiferous epithelium showed degenerative changes.   At the  6 week



time point, these  degenerative changes  were more  pronounced,  and there was a



complete absence of  spermatocytes  in  the  lumen.   Cr(VI)  produced mild edema of



the interstitial tissue of the testes at  3 weeks.  Following 6 weeks of exposure,



edema was more marked  and  congestion  of the blood vessels was noted. Although



the seminiferous epithelium appeared  normal, the tubules  were devoid  of sperma-




tocytes .



     Hopkins  (1965)  postulated that   incorporation  of chromium into sperm and



subsequent  passage  into the  epididymis  may be a  problem.   This  hypothesis is



based upon  observation of chromium uptake by testis, subsequent decline of  tes-



ticular  radioactivity,  and   subsequent  increase  in  radioactivity  in  the



epididymis  (see  Section 5.2.2.).    Gross  and  Heller   (19^6)  have reported



sterility in rats exposed  to  zinc  chromate and potassium chronate in the  diet




(Section 7.2.5.) .



     7.2.3.3   SWWARY — Chromium has adversely affected fetal development and



male  reproduction in  experimental animals.   Hamsters  administered chromium



trioxide intravenously on day 8 of gestation had an increased incidence of cleft
                                     7-111

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palates in the young when examined on  day  15 of gestation.  The  malformations




were strain specific and associated with maternal  toxicity.  Maternal  toxicity




was  not observed  in  the strains  where fetal  malformation was  not  present.




Increases  in  external  malformations  also were  observed  in  mice, following




subcutaneous administration of CrO_.




     Other reproductive effects of chromium  include  testicular  degeneration  in




rabbits receiving 2 mg/kg/day for 6 weeks  of  either Cr(III) or  Cr(VI) compounds




by intraperitoneal injection.   The Cr(III) compound produced more severe effects




in this study than did the Cr(VI) compound (Behari et al.,  1977).   The relevance




of  these observations  to  effects observed  after  environmental  exposure are




questionable, since the route of exposure  was not natural.








7.2.M.    Chromium Hypersensitivity.








     7.2.1.1.  CHROMIUM SENSITIVITY AND CONTACT  DERMATITIS  —  Chromic  acid and




the  chromates  are powerful  skin irritants,  and,  in lower  concentrations, the




chromates are sensitizers (NAS, 197*O.   Workmen  exposed  to  the  steam of boiling




dichrouate solutions  developed an acute  primary  irritant contact  dermatitis




(Schwartz et al., 1957;  White,  1931*).  White (193*0 described a diffuse erythema-



tous dermatosis that resulted  from dichromate; seme  progressed to an exudative




phase.




     Various chromium compounds have  been  implicated in  giving  rise  to  allergic




dermatitis with varying  degrees of eczema.   Parkhurst (1925) reported the case  of




a woman employed in blueprint  production using a process in which a 156 potassium




dichromate solution was  used as a fixative.  A 0. 5% potassium dichromate  solution




was rubbed on the right  thigh of the woman, and soom after the application,  there




was a local sensation of itching and  burning.  Twelve hours later,  the  patient
                                     7-112

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developed  a  follicular  erythematopapular  deraatitis  at  the  exposure  site.


Itching and burning was reported when a similar application was made to the left


thigh.

     Smith (1931)  reported a case of chromium sensitization in a man who had been


hospitalized after occupational exposure  to ammoniun dichrcmate.   The  patient


had ulcerations in the  skin of both hands, and complained of asthma, and muscular


weakness and tenderness.  He had  a previous history of  asthma and  hay fever and


had further asthmatic attacks  upon exposure to chromium.  Following a patch test

                                                P
with  1$  ammonium  dichromate solution on a  1 cm   area of normal  skin on  his


forearm, the man  developed a  mild erythema after  24 hours.   After  3 days,  the


erythematous area doubled in size,  and there was the appearance of  vesicles.   An


intradermal injection of 0.1  m2,  of a 0.5% aqueous  solution of ammonium dichro-


mate was given in the right forearm 8 days  later.  Within  an hour,  the patient


developed  a  generalized pruritis  with  soreness at  the  injection site.   This

progressed with the development of a vesicular erythematous dermatitis covering


the  entire hands  and lower parts  of the  forearm.   In addition, he developed


diaphoresis and sibilant  rales.  These symptoms which  required  hospitilization


abated after  his  exposure to Cr(VI)  ceased.  Three control  subjects  who were


similarly injected showed no reaction.

     Hall  (19UU) reported on 132 aircraft workers who developed dermatitis after


contact  with a primer  consisting of a suspension  of zinc chrcmate  powder and

magnesium silicate in a xylene solution of certain resins, including a  phenol-


formaldehyde resin.  Those workers  who  had dermatitis from  the primer and who

were  allergic  to  zinc  chromate  pigment   had  a  mean  duration of  employment of


7 months (range:  1 week to 9 years).  A battery  of  patch tests showed that 90 of


the workers (68$)  were sensitive  to the zinc chromate pigment only. (Apparently,


the zinc chromate pigment was  a mixture of zinc chrcmate and calcium carbonate.)
                                     7-113

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Only a few workers had positive patch  tests  for other compounds encountered on



the job.



     Forty-five cases of  allergic  contact  dermatitis, observed in the Helsinki



area from  1945  to 1918,  were reported  by  Pirila and Kilpio  (1949).   Positive



patch-testing with a 0.5%  aqueous solution  of potassium dichromate (pH 4.15) was



observed in  41  of the  workers.    The  workers  were  involved in  the  following



occupations:    11 bookworkers,  10 cement  and  lime  workers,   7 radio  factory



workers using a photostatic procedure,  4 metal  factory workers, 4 painters and



polishers, 3 fur workers,  and 6 others.




     Engebrigsten (1952) reported eight cases of cement eczema among 300 to 400



Norwegian workers exposed "more or less directly" to cement dust that contained



0.002 to 0.020% water-soluble  Cr(VI), which was  described only as "water-soluble



chromates."  Positive patch testing was reported in  7 of  8 patients exposed to



0.5$ aqueous  solution  of  potassium dichromate.   Positive reactions  to cement



patch tests were observed in 4 of 8 patients.   Of the 10  persons  who  served as




controls, none  gave  any positive  reactions.  Engebrigtsen (1952)  subsequently



tested the same eight patients with a cement slurry that had been washed free of



Cr(VI), and none of the people reacted positively.



     Denton et al. (1954)  patch tested a patient with a "strong specific hyper-



sensitivity to potassium chromate"  with three solutions:   (1) a 50 ppm solution



of potassium dichromate,  (2)  1 ppm water-soluble Cr(VI) filtrate  from American



Portland  cement,  and  (3) 4 ppm water-soluble  Cr(VI) filtrate from  American




Portland cement.  Each patch  test  resulted in  an erythematous,  edematous, and



papulovesicular reaction.   There was  no reaction to  distilled water and none of



the control subjects  reacted to any of the  three Cr(VI) solutions.




     Six out of 200 employees  who worked in a diesel locomotive repair shop were



incapacitated by chromate dermatitis (Winston and Walsh, 1951).  All were exposed
                                     7-114

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to an  alkaline  diesel locomotive  radiator  fluid that  contained  0.08$ sodium



dichromate.   Positive  patch testing was  reported for  both sodium dichromate



(pH 4.25) and the radiator fluid  (pH 10).



     Walsh (1953),  in a summary report on chromate hazards in industry, described



the following patch test results:   2%  "chromate  acid"  applied for 24 hours on



superficial skin abrasions  produced a  crusted lesion  in 3 weeks;  0.5% sodium



dichrcmate, reapplied daily for  3  days, produced  a  crusted lesion in 3 weeks;



0.5% potassium  chromate,  applied  8 hours/day for 3  days,  produced lesions in




3 days; 0.5% sodium chronate,  0.005$ sodium dichromate,  and pure zinc chrcmate



also produced lesions in 3 days after being in contact with skin for 8 hours/day



for 3 days.  Lead  chronate did not  produce a  reaction  after the same exposure



period.  A 10$ solution of Cr(III)  nitrate produced  redness after the solution



was reapplied daily for 3 days.



     Edmundson (1951) obtained only two positive reactions with patch testing in



56 men who had  chrome  ulcers.   All men were said to have  a history of chrome



dermatitis.  He concluded that the  presence of chrome ulcers did not necessarily



indicate sensitization.



     Other investigators  (Morris, 1955; McCord et al., 1931; Levin  et al., 1959)



have demonstrated sensitization  to  other chromium  compounds  in  workers employed



as tanners and  lithographers.   Many of the workers  gave positive results for



patch tests.



     There have been numerous  case  reports of  contact dermatitis resulting from



exposure to a variety  of  chromium-containing  products,  such as matches, auto-



mobile primer  paint, and  fumes  from  welding rods   (Fregert,  1961;  Engle and



Calnan, 1963; Newhouse,  1963; Fregert and Ovrum,  1963; Shelley, 1964).  In most



of these reports, the subjects developed a positive reaction to patch testing for



a solution of potassium dichrcmate.
                                    7-115

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      A study conducted in France by Jaeger and Pelloni (1950) demonstrated that



 workers with cement eczema were sensitive  to  potassium dichronate.   In their



 study, the  authors  patch tested 32 patients with cement eczema and 168 patients



 with eczema from other causes.  Those  with cement eczema  gave  positive patch



 tests (9W  to an aqueous 0.5% solution  of potassium dichromate, while only 5% of



 the other eczema patients exhibited positive reactions from the dichrcmate.



      However,  Perone et al.  (1971*)  reported  that  only 2  of 95  construction



 workers who regularly worked  with cement  gave  a positive reaction  to  patch




 testing to  a solution of  0.25$  potassium dichromate or a  solution containing



 H50 ng/g Cr(VI) extracted  from cement.   The authors suggested that  the  cement



 dermatitis  present  in the  construction  workers was associated with the initia-



 tive nature  of the  cement rather than a hypersensitivity.



      There  have  been two reported  cases of eczema in men who regularly worked



 with cement and who each  had a  green tattoo which contained  chromium (Cairns  and



 Calnan,  1962; Loewenthal, 1960).  Both men developed a positive reaction to patch



 testing to a solution of  potassium  dichrcmate (concentration range:  0.1 to 2%).



 Negative results  following patch testing were observed  when the  solution  was



 Cr(III) sulfate.  The green pigment in  one  of the tatoos contained Cr(VI),  but



 the  oxidation state of chromium in the other tattoo was  not  determined.



     Krishna  et  al. (1976) reported  that chrome  workers  exposed  to  0.21   to



 0.80 mg/nr  Cr(VI)   displayed  various  dermatologic  disorders,  including skin



 ulcers,  dermatitis,  and  nasal  perforations.   Dermatologic  irritation  has been



 reported by  Clausen and  Rastogi  (1977), in auto workshop employees  exposed  to



 0.01 to 3.75 \ig/nr; by Tan don  et  al.   (1977),  in  electroplaters,  polishers,  and



pigment workers; and by Spruit and Martin  (1975), in offset  printing employees



who  came in  contact  with materials  (usually inks)  containing up to  60,000  ppm



chromium.   Various dermatologic disorders  among workers  exposed  to  0.001   to
                                     7-116

-------
 0.020 rag Cr/m  in  air and chromium on work surfaces have been reported by Lucas



 and  Kramkowski   (1975).    Chromium-induced  dermatitis  or  eczema  has  been
                                                *


 associated  with  workers  having  contact  with  various  chromium-containing lubri-



 cants  and  oils  (Rosensteel  and Lucas,  1975;  Weisenberger,   1976;  Clausen  and



 Rastogi,  1977).    Dermatitis  associated with  various  chromium  pigments  and



 coloring  agents  has  been  described by Scmov  et  al.    (1976), Fisher  (1977),



 Fregert  and  Gruvberger  (1976),  Venediktova   and Gudina  (1976),  and  Evans



 (1977a,b,c).



     Allergic  sensitization  to  chromium-containing compounds  has  been reported



 by  Burry  and Kirk (1975) in  several  industrially exposed individuals,  and  by



 Wahlberg and Wennersten  (1977)  and Kaaber and  Deien  (1977), who used potassium



 dichrcmate  in  the  patch tests.   Husain (1977)  showed  sensitization  in  the



 "general population"  (not  occupationally exposed), where, among  1312 patients



 tested, 11.58? had contact sensitivity  to 0.5$  potassium dichronate  in  a patch



 test (15.59$ of  the males reacted to  the chromate as opposed to 8.18$ of  females



 tested).



     Perisic and Jovovic (1977)  have  identified allergic contact sensitivity to



 chromates among  219 Yugoslavian housewives.   The incidence of allergic  contact



 sensitivity was confirmed in  43  cases (19.62$);  the percentage  of positive cases



was 31.44$ in women holding  permanent  jobs,  23.51$ in cleaners, and  14.58£  in



unemployed housewives.  It was concluded by the authors that chromates contained



in a variety of  household products used  in routine cleaning operations  are  the



cause of allergic  sensitivity to  chromates  in women.   No control or reference



data were available.



     Jovovic et  al. (1977) have confirmed the  allergic  contact sensitivity  of



shoemakers  exposed to  chrcmates  in occupational  settings.  Sixty percent  of the



shoemakers  with  allergic sensitivity showed a  positive reaction  to  potassium
                                    7-117

-------
dichrcmate.  Allergic reactions in various  glues  to which the shoemakers were



exposed were also reported.



     Exposure  to  chromium may  sensitize  certain  individuals  resulting  in



asthmatic attacks upon subsequent re-exposure (NAS, 1 97*0 (see Section 7.2.5.1).



     7.2.4.2.  SENSITIVITY TO CHROMIUM IN PROSTHESES — In recent years, there



has been  an  increase  in the use of  metal-to-pi as tic  prostheses in  orthopedic



surgery.   Many  of the  metal  implants  consist  of  alloys  containing nickel,



chromiun, cobalt, and molybdenun.




     Work by Swanson et  al. (1973) and Coleman et  al. (1973)  has  shown that when



two cobalt/chrome  alloy surfaces rub together, cobalt  and chrome are released



locally, pass into  the blood and circulate throughout the tissue of the body, and



are finally excreted in  the urine.  In patients sensitive  to metal, the vessels



supplying the  bone in which the prosthesis  is  inserted may show obliterative



changes, and in theory, lead to  death  of the bone,  with weakening  of the fixation



between the bone and prostheses, and  consequent loosening  (Anonymous,  1976).



     There is considerable conflicting evidence in this  area, as Deutman et al.



(1977)  reported  metal sensitivity  in H of  66 patients  induced  by metal alloy



implants following total hip replacement arthroplasty;  however,  positive patch



tests  occurred only with  cobalt  and  nickel,  while tests  with chromium were



negative.  Brown et al.  (1977)  gave patch tests for chromium sensitivity to 20



patients with sterile loose hip replacements.  Of these 20 patients,  none were



positive for chromium sensitivity,  and of 17 patients that were reoperated on,



none showed histologic evidence of  delayed hypers ens itivity around the implant.



     In  addition,  contact  dermatitis  and  allergic sensitivity  to   chromium-



containing  dental   prostheses  have  been  reported  by  Levantine  (1974)  and



Ovrutskii (1976).
                                    7-118

-------
     7.2.4.3.   ANIMAL STUDIES ON  CHROMIUM SENSITIVITY — Nunerous investigators


have demonstrated that sensitization of  laboratory  animals  can be produced by


exposure to various chromium compounds, including both Cr(VI) and  (III).


     In studies  of delayed  dermal  hyper-sensitivity to potassium  chromate or

                                                         _ii
chromium  chloride,  guinea  pigs  treated  with  2.4 x 10   M  chromic  sulfate


developed signifiant  cross sections (Gross et  al.,  1968).  The ability  of Cr(III)


compounds  to  evoke  an  allergic  reaction  in chromium-sensitive  guinea  pigs


decreased gradually in the following order:  chromic chloride  > chromic nitrate >


chromic sulfate > chromic acetate > chromic oxalate (Scheiner  and Katz,  1973).


According to the authors, this difference  depended on the free Cr(III)  concentra-


tion in the solution which, in turn, depended on the degree to which the Cr(III)


formed coordination complexes with the  original ligands  present; strong ligands


prevented the  formation of  complete antigens.  Sensitization  of guinea pigs  with


chromium (III) sulfate  (using Triton X-100) was 86$ successful,  whereas sensiti-


zation with an aqueous  solution of potassium chromate was successful in only %%


of the animals tested (Schwartz-Speck and Grundsmann,  1972).


     Jansen and Barrens (1968)  reported that  guinea  pigs could  be sensitized to


both Cr(III) and Cr(VI) by the subcutaneous injection of aqueous solutions  of the


appropriate chromium salts in a 1:1  emulsion  with Freund's complete adjuvant.


     Hicks  et al.  (1979) produced hypersensitization  in  guinea pigs  with  both


Cr(VI) and  (III)  salts.    The  contact  sensitization potentials of the Cr(III)


complexes in guinea pigs  were proportional to the release of chromium from the


complex (Schneeberger and Forck,  1974) .




7.2.5.    Other  Toxic Effects  of  Chromium.   The  literature concerning chronic


and subacute exposure to chromium consists primarily of reports  of no  observable


effect levels  (NOEL) .   Although  the  kidney and  liver  have been  shown  to be
                                     7-119

-------
targets  following acute  exposures,  reports  of  pathology  in  these  organs



following long-term exposure via relevant  routes  were not  found.   There are two



reports which suggest effect levels following oral exposure; these studies are



summarized  in  the following  text.    The  numerous studies  which  suggest free



standing NOAELs are of limited value  as  a  result  of experimental design and are



summarized in Table 7-20.



     Gross and Heller (1946) reported  that 0. 125/K K^rCL in the feed of rats was



tolerated without observable effects,  0.25% resulted in "subnormal condition",



including rough coat  and "subnormal" young born to treated animals.  Doses of 0.5



and 1$ resulted in diarrhea, rough dirty coats, and sterility.   ZnCrOj. adminis-



tered in the feed at  levels of 0.125,  0.25,  0.5,  and  1.0$ resulted in subnormal



appearance, rough  and  dirty coats,  and sterility  at  all  dose  levels.   Group



sizes, duration of treatment,  and criteria  for determining sterility were not



reported.



     Ivankovic and Preussman  (1975) administered  Cr 0   to rats in their feed.



The compound was prepared by incorporating it into bread dough  at levels of 2 or



5%.   In a  2-year  study  (600  feeding days), it  was  estimated that  for the 2%



treatment level,  males  consumed  75 g/kg  (25 g/animal)  over  the  entire  study



period and  females consumed 72 g/kg  (18 g/animal);  at  the 5% treatment level,



males consumed  180 g/kg and females 160 g/kg  for the duration of the study.  Body



weights were monitored and  animals were maintained on normal  rations following



the treatment  period.  At death, animals were autopsied and "all the important



organs" were studied  for micro pat ho logy.  In  addition, a 90-day study was conduc-



ted also using bread  levels of 2 or  5%.  In  this  study,  urinary  protein, sugar,



bilirubin, blood, and sediment were monitored.  During the last 30 days of the



study, treated  animals were mated and the number of viable young produced in each



litter was recorded.   At the end  of treatment, blood samples  were analyzed for
                                     7-120

-------
            TABLE  7-20

Studies Suggesting NGAELS or NOELS
Species
mouse


mouse

rat
(young)

rat
(young)
-•4
1 rat
KJ

dog


Route Con pound
drinking K0CrO,.
water 2 M

feed aiCrO,,

drinking fC_CrO,,
water 2 "

feed K-CrO,.
£. *T

feed Cr 0-
2 3

drinking K,CrO,.
water * ^

Dose Duration
100, 200, 300, N. S.
300, MOO, or
500 ppra
1) N. S.

300 and 500 ppm N. S.


0. 125J N. S.


0, 1, 2, or 9)t 2 years


0. 45, 2.25, 4 years
1.5, 6.75,
or 11.2 ppm
No. at End point a
Start Monitored
N. S. general appearance ,
reproduction

N. S. general appearance ,
reproduction
N. S. general appear an 09 ,
reproduction

N. S. general appearance ,
reproduction

60 / gross and micro-
group acopic pathology,
body weights
2 /group urinalysia Including
albumin, acetone,
bile plgnents
Result Reference
NCEL for all Gross and Heller, 19t6
doses

NCEL Gross and Heller, 19t6

NOLEL; slight Gross and Heller, 1946
roughness of
coal at 500 ppm
NCEL Gross and Heller, 1946


NCEL I van to vie and Preussman,
1975

NCEL Anwar et al., 1961


                   glucose, indican
                   erythrocytes , and
                   specific gravity;
                   gross and micro-
                   scopic pathology of
                   adrenals, bone
                   marrow, brain, heart,
                   intestine, kidney,
                   liver, lung,
                   mesenteric lymph
                   node, parathyroid,
                   pancreas, spinal cord,
                   spleen, stomach,
                   thyroid, and tonsils;
                   weights of liver
                   spleen and kidney

-------
                                                               TABLE 7-20 (cont.)
No. at
Species Route Conpotnd Dose Duration Start
rat drinking K_CrOtt 0, 0. U5, 2.2, 12 mo highest
water 1.5, 7. 7, 11 dose 9
and 25 ppm females ,
12 males
control
10 of
each sex,
all other
groups , 8
males, 8
females
End points
Monitored Result Reference
clinical blood NOEL MacKenzie et al., 1958
chemistry, body
weights gross and
microscopic patho-
logy






rat
cat
drinking    CrCl,
water           J
            0,  25 ppm
feed
chroniun    50 to  100
carbonate,  mg/ oat/ day
chromlun
phos phat e
cat
             inhalation
            chromium
            carbonate
            dust
            from 3.3 to
            to 83  mg/nr
            average  58. 3
            mg/nr
12 mo       12 males,  clinical chemistry     NOEL
            9 females  body weights, gross
                      and microscopic
                      pathology

1 to 3      10         organ weights          NOEL
mo                    macrosoopic patho-
                      logy microscopic
                      pathogy of lung,
                      heart, liver, stonach,
                      spleen, pancreas,
                      kidney, brain,
                      sketal muscles

86 see-     2          gross and micro-       NCEL
slons                 scoplc pathology
which
varied from
10 to 60
min and
a vera ged
28 mln
for one
cat and
57 min
for the
other
                                                                                                                     MacKenzie et  al.,  1958
Akatsuka and Fair hall,
193t
                                                                                                                     Akatsuka and Fair hall,

-------
 blood sugar,  serum  protein, serum bilirubin, and haemoglobin.  Erythrocytes and




 leukocytes were counted.  Liver, spleen, kidney, brain, and ovaries were weighed,




 fixed, and sectioned  at  autopsy.   In addition, lung, heart, pancreas, stonach,




 small intestine, and  urinary bladder were also fixed and sectioned.




      In the  90-day  study, the only treatment related effect was a depression of




 spleen and liver weights.  Spleen weights appeared  to be depressed at both doses




 in  both sexes.  Liver weights appeared to be  depressed in both dose groups for




 females and in high dose males.  Statistical analyses were not  reported.  In the




 2-year study, no treatment related effects were reported.




      7.2.5.1.  RESPIRATORY  EFFECTS  —  Steffee  and  Baetjer  (1965)   exposed




 rabbits,  guinea pigs, rats, and  mice to mixed chromate dust  via inhalation  or




 intratracheal  injection.   Inhalation  exposures  were conducted  5 hours/day,




 4 days/week throughout the lifespan of the animals.  The average air concentra-




 tion  was  estimated to be 3 to 4 mg of CrO_/m .  The average weekly  exposure was




 estimated  to  be 53,  44, and 49 mg/hour  for rabbits, guinea  pigs, and  rats,




 respectively.   The  following exposure related  effects were documented in the




 lungs of exposed animals.   Fifteen percent  of the  rabbits  and  rats  in the




 inhalation exposures, and  rabbits  and  guinea pigs  in  the  intratracheal groups




 exhibited granulcmata. This lesion was  found in only  one control rat and in none




 of  the rabbits or  guinea pigs.   The  incidence of  alveolar  and  interstitial




 inflammation was greater than controls in guinea pigs  exposed  via  both  routes.




 Exposure related effects in liver,  kidney, and spleen were not  found.




      Nettesheim et  al. (1971) exposed mice to calcium chrcmate dust  5 hours/day,




 5 days/week for life.   The exposure concentration was 13 mg/m^  of CaCrCL.   This




exposure concentration depressed weight  gains.   Exposed  animals showed marked




alterations in the  epithelium  of  the  bronchial  tree after  6 months of exposure.




These  alterations were  graded  from epithelial necrosis and atrophy to marked
                                    7-123

-------
hyperplasia.  In addition, bronchiolization of alveoli  (bronchiolar cells lining



alveolar walls) was  observed.   Another frequently observed effect was alveolar



proteinosis.  In  addition to  lung effects, morphological changes were noted in



the  tracheal  submandibular lymph  nodes.   After 2 years of exposure, spleen and




liver were  atrophied.   Ulcerations in the stomach and  intestinal  mucosa were



noted occasionally.




     Early historical recognition of the ulcerative property  of Cr(VI) compounds



in   humans  is  evidenced  by  several  studies  on the   subject  (BScourt  and



Chevallier, 1863; Delpech  and Hillairet, 1869; Legge,  1902).



     Bloomfield  and  Blum  (1928)  reported on  23 men  employed  in six chromium



plating plants in the United States. Their findings are presented  in Table 7-21.



They concluded that  continuous daily exposure to chromic acid at  concentrations



>0.1  mg/nr*  is  likely  to  cause  nasal  tissue  injury.   As  can  be seen  from



Table 7-15, no concentrations <0.12 mg/nr were observed;  hence, injury to nasal



tissue caused by lower concentrations could not be ruled out.




     Four of 33 chromium platers were  found to have septal perforation, although



the  highest measured concentration of  chromium trioxide in the  workplace was



0.003 mg/m .  Of the 33 workers, six had what  the author  considered to be normal



nqses.  He suggested  that in view  of the low chromium concentration,  the lesions



that resulted were due  to exposure to periodic high concentrations  of chromium



trioxide that occurred when ventilation of the tank failed (Lumio, 1953).



     Anodizing operators exposed to concentrations of  chromic acid mist ranging



from 0.09 to 1.2 mg/m  (as CrO ) developed ulceration  of the nasal passages and
atrophic rhinitis (Gresh, 19^; Zvaifler,



     The United States  Public  Health Service conducted  a study on  workers in




seven chromate-producing plants  in  the early 1950s.   The results  are shown in



Table 7-22.   Unfortunately,  the  results of the  physical examinations  on the
                                     7-124

-------
                           TABLE 7-21




Clinical Findings in Workers Employed in Chromium-Plating Plants
Case
1
2
3
it
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
Time Employed in
Chromiun-Platlng Time Over
Occupation Room, mo Tank, h/d
Chromiun plater
Chromim plater
Foreman plater
Foreman plater
Chromiun plater
Chromiun plater
Chromlun plater
Chromiun plater
Chromlun plater
Chromlun plater
Chromiun plater
Chromlun plater
Chromlun plater0
Chromiun plater
Nickel platerd
Backer
Backer
Backer
Wiper
6
20
7
8.5
3.5
0.75
0.25
7
3
36
5
0.75
12
0.67
1.5
8
0.75
0.75
1.5
U
4
2
3
4
7
7
7
7
4
6
6
4
2
0
0
0
0
0
Approximate
CrO. Expostre, Perforated Ulcerated
mg/m Septun Septun
1.5 ++
2.8 -H.
2.5 - -M-
2.5 - *+
5.6 - -M.
0.12
0.12
0.12
0.12
0.2
0.12
0.12
2.8
2.8
?
?
?
?
?
Inflamed
Mucosa" Nosebleed
•M. Yes
+ Yes
++ Yes
++ Yes
-M- Yes
•M. Yes
•M- Yes
++ Yes
++ No
++ No
+ Yes
+ No
No
No
+ Yes
+ Yes
+ No
+ No
+ No
Chrome
Holes
Yes
Yes
No
No
Yes
Yes
No
No
Yes
No
Yes
No
No
No
No
No
No
No
No

-------
                                                                  TABLE 7-21 (cont.)


1
ON

Case
20
21
22
23
Occupation
Foreman6
Foreman6
Clerk6
Inspector6
Time Employed in
Chromlun-Plating
Room, mo
0
0
0
0
Time Over
Tank, h/d
0
0
0
0
Approximate
CrO, Exposure, Perforated Ulcerated
mg/m Septun Septun
0 -
0 -
0 -
0 -
Inflamed
Mucosa Nosebleed
+ No
+ No
No
+ No
Chrome
Holes
No
No
No
No
^Soiree:   MAS,  1974a
c++, marked; +,  alight; -, negative.
 .Used vaseline In nose.
 Cyanide  burns
 Worked in other  departments of factory
mo = month;  h = hoir;  d = day

-------
—I
 I
                                                       TABLE 7-22


                                     Perforation of Nasal Septum in Chrcmate Workers*
All Workers
Time Worked
in Chroraate
Industry
<6 months
6 mont hs to
3 years
3 to 10 years
>1 0 years
TOTAL
Total
No.
41
117
370
369
897
Workers wi th
Perforation
No. %
1 2.4
46 39.3
205 55.4
257 69.6
509 56.7
White Workers
Total
No.
32
89
235
297
653
Workers with
Perforation
No. %
0 0
28 31.5
104 44.3
190 64.0
322 49.3
Nonwhite Workers
Total
No.
9
28
135
72
244
Workers with
Perforation
No.
1
18
101
67
187
*
11.1
64.3
74.8
93.1
76.6
          •Source:  NAS, 1974

-------
workers were  not  related to chromiun  exposures,  and hence, the  data are  of




limited usefulness (Federal Security Agency, 1953).




     Mancuso (1951)  reported  on  physical  examinations  of  a  random  sample  of  97




workers from a chromat e-eheraicaT  plant.  It can be seen from the results  which




are presented in Table 7-23, that 61 of  97 workers ( 63%) had septal  perforation.




The data suggested to the author that Cr(III) may be partly responsible for the




perforations;  however, later  studies have not provided support for  this theory.




     The results of examinations  of nine  workers  in a chrome-pi at ing plant are




shown in Table 7-24.  Analyses of air samples showed chromiun concentrations  of




0.18 to 1.4 mg/m .  Sane degree of nasal septal uloeration was seen  in 7 of the 9




men, with 4 of 7 demonstrating frank perforations  (Kleinfeld and Russo,  1965).




Unfortunately, the effects of chromium  for a specific length of time at a  fixed




concentration were not studied.




     In a  Russian study  conducted  by  Kuperman  (196*0,  10  apparently normal




persons were exposed to Cr(VI)  aerosol  concentrations of  non-reported composi-




tion ranging  from 0.0015 to 0.04 mg/m.   Air containing  Cr(VI)  at 0.01  to




0.024 mg/nr sharply irritated the  nose  when inhaled  for short periods of  time.




The most sensitive  person responded  at a  chromiun  concentration  of 0.0025  to




0.004 mg/m ; however, it  was  not known  if this was a reaction to chromiun  or  to




the acidity of the aerosol.



     Vigliani  and  Zurlo   (1955)  reported nasal  septal perforation  in workers




exposed to chromic acid  and chromates in  concentrations of  0.11 to 0.15 mg/m .




The lengths  of exposure  were not  known.   Otolaryngologic  examinations  of  77




persons exposed to chromic acid aerosol  during chrome  plating  revealed 19$  to




have septal perforation and  48$ to have nasal mucosal irritation.  These people




averaged 6.6 years of exposure to an air  chromiun concentration of 0.4 mg/m .  In




14 persons, papillonas of the oral cavity and larynx were foind.  The diagnosis
                                    7-128

-------
                                     TABIJE 7-23

                  Perforation of Nasal Septum in Chronate Workers*
Ratio of
insol Cr*^ to Chromium Concentration,
sol Cr mg/nr (as Cr)
Workers in plant
<1.0:1 <0.25
0.26 to 0.51
£0.52
1.1 to 4.9:1 <0.25
0.26 to 0.51
>0.52
>5.0:1 <0.25
0.26 to 0.51
>0.52
TOTAL
Office workers 0. 06
No. Workers
Examined

4
7
8
9
32
15
7
2
13
97
4
Workers
Septal
No.

2
3
4
7
20
11
2
1
11
61
0
with
Perforation
%

50
43
50
78
63
73
29
50
85
63
0
•Source:   MAS,  1974
insol =  insoluble; sol = soluble
                                     7-129

-------
                                     TABLE 7-24




                 Nasal  Medical Findings  in a C hromi un-P lat ing Plant*
Case
1
2
3
4
5
6
7
8
9
Age, yr
30
19
19
18
47
U5
23
20
48
Duration of
Exposure, mo
6
2
12
9
10
6
1
0.5
9
Findings
Perforated septan
Perforated septun
Perforated septum
Perforated septum
Ulcerated septum
Ulcerated septun
Ulcerated septun
Moderate injection of septun and turbinates
Moderate injection of septun
"Source:   MAS,  1974
                                         7-130

-------
 of  papillona was  confirmed  by  histologLc  examination.   There were no signs of


 atypical growth or malignant degeneration (Hanslian et  al.,  1967).


     Cohen et al. (197U) have identified a serious health hazard among workers in


 a nickel-chrome plating area.   Thirty-five of  37 (95/6)  employees exposed to

                                                •3
 atmospheric concentrations averaging 0.0071 mg/m  as total chromium were shown


 to  have developed significant nasal  pathology and skin lesions characteristic of


 exposure to chromic acid.  The authors  attributed the  high incidence  of adverse


 health effects to the lack of emphasis  on the  implementation  of good industrial


 hygiene and personal hygiene.  The mechanism  postulated for the occurrence of the


 observed nasal damage resulted from either long-term exposure to levels of Cr(VI)


 below prescribed "safe" levels, or  direct contact of the  affected tissue resul-


 ting from inadequate personal hygiene practices.


     The literature suggests that chromiun compounds are responsible for a wide


 variety of  other respiratory effects.   Studies done  by  German  investigators


 demonstrate mixed results from exposure to  chromiun compounds.  Fischer (1911)


 and Lehmann  (191U)  reported  that  there  were  no marked  clinical  symptoms  in


 persons exposed to chromate  dust.  Other German investigators  (Alwens and Jonas,


 1938; Fischer-Was els,  1938;  Koelsch, 1938;  Lehmann, 1932; Mancuso, 1951)  have


 reported that  prolonged inhalation of chrcmate dust  caused  chronic irritation of


 the respiratory  tract and  resulted  in such manifestations  as congestion  and


hyperemia, chronic  rhinitis,  congestion  of the larynx,  polyps  of  the  upper


 respiratory tract, chronic  inflammation of the  lungs, emphysema,  tracheitis,


chronic bronchitis,  chronic  pharyngitis, and broncho pneumonia.  X-ray findings


included enlargement  of the hilar region (often on only one side), enlargement of


the lymph nodes, increase in perlbronchial and peri vascular lung  markings ,  and


adhesions  of the diaphragm.   Letterer et al.  (197*0 and Lukanin (1930) stated


that a characteristic pneunonoconiosis resulted from  exposure to some chronates.
                                    7-131

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With all of these studies, a correlation between symptomatology, physical signs,




length of exposure, and dose of chromium compounds was not available.




     In  contrast with the above  findings, 897  workers  in  chromate-producing



plants in the United  States had a higher  incidence of severely red throats and




pneumonia, but did not show any increase  in the  incidence of other respiratory




diseases when compared with control groups.   Although bilateral hilar enlarge-




ment was observed, there was no evidence of excessive pulmonary fibrosis in these




workers  (Federal Security Agency,  1953).  The various lung changes described in




these workers may  represent a nonspecific  reaction to irritating material  or a




specific reaction to chromium  compounds.  Many of the conditions mentioned occur




widely in the general adult population (NAS,  197U).




     Inhalation  of massive amounts  of chromic acid mist  has  resulted in acute




pulmonary complications.  Meyers (1950) reported on two patients who were exposed



to an  estimated  chromic  acid mist  concentration of 20 to 30 mg/m .   Symptoms




included  cough,   dyspnea,  chest   pain,  and weight  loss.   The first  patient




developed pulmonary  edema,  and the  author stated  that his exposure  was  more




severe and  prolonged  than the second patient who  developed a  slight pleural




effusion.  In another study,  Zvaifler (19^)  reported atrophic rhinitis in 5 to




10$ of a group  of workers exposed to  the  mist  of a 5% chromic acid solution.




Hyperemia,  congestion,  swelling,  and nasal   inflammation  recurred in  some  of



these persons.




     Gomes  (1972)  examined  303 employers  in  81  electroplating  operations  who




worked in Sao Paulo, Brazil.  Over two-thirds of the workers had mucous membrane




or cutaneous  lesions,  with  many  of  them  having ulcerated or  perforated nasal




septa.  The duration of exposure was not stated,  but  the  author mentioned  that




the harmful  effects  were noted  in less  than 1  year.   In addition,  a direct
                                     7-132

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correlation  between  workers  exposed  to  a  given  airborne  concentration  of


chromiun (VI) and the  development  of harmful  effects could not be made.


     Cohen and Kramkowski  (1973)  and  Cohen  et al. (1971) examined  37 workers


employed by  a chromium-pi at ing plant.   Within  1  year  of being  employed,  12


workers experienced nasal  ulceration or perforation. The  airborne chromiun (VI)


concentrations ranged  from <0.71 to 9.12 M.g/m .


     In a chromiun plating plant where the maximun airborne chromiun (VI) concen-


tration was  3 ng/m ,  no ulcerated nasal mucosa or perforated nasal septa were


found; however, half  of the 32 employees had  varying degrees  of mucosal irrita-


tion  (Markel and Lucas, 1973).  This  was  not thought  to be  significant by the


investigators, because the survey was carried  out at the peak of the 1972 to 1973


influenza epidemic. The length of employment  for the workers was as follows:  >8


years, 15 workers; 4  to 8  years, 7 workers; 1 to  4  years, M workers; <1 year, 6


workers.


     Machle and Gregorius  (1948) reported  an  incidence of nasal septal perfora-


tion  of  ^3.5%  in  354  employees who  worked in  a  chronate-producing  plant that


manufactured sodium chromate and bichromate.  At the time of the study, airborne

                                                    •3
chromate concentrations ranged frcm 10 to 2800 jig/m .   The  plant has  been  in


operation for at least  17 years, and some employees had probably worked in the


plant when reverberatory furnaces, a prominent source of  high  chrcmate exposure,


were used.


     Chromiun exposure  in Australian  shipyards,  resulting from welding opera-


tions, has  been  reported  by Bell  (1976)  to  cause irritation to the  nose and


throat at welding fume ranging from  0.006 to  0.05 mg/  .  Lung biopsy specimens


from welders  confirmed  the  presence  of severe  pneunoconiosis  similar  to that


reported by Stettler  et  al.  (1977).
                                    7-133

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     Various  other  disease  states  such  as  asthma  have  been  attributed  to



chromium,  but, in most cases,  the  etiologLc  relation to  chromium  is doubtful



because of the presence of other chemicals (MAS,  1974).  Asthma was reported as a



complaint among workers employed for >10 years in a ferrochromium plant in Norway



(Broch, 1950).  Bronchial asthma was  also reported in Russian bauxite workers,



where Cr(VI) was involved in  the bauxite caking  process  (Budanova,  1976).   A



clinical test for the diagnosis of actual or imminent bronchial  allergic reac-



tions to Cr(VI) has  been  developed by  Budanova and Makarova (1979).   A correla-



tion was  established between  the  concentration of  leukocyte agglomerates  in



blood stimulated by Cr(VI) ,  with  3% for a  healthy  control group versus =16$ for



workers with heavy bronchial asthma.  It appears, then, that chromium sensitiza-



tion can occur  after inhalation  exposure  as  well as  dermal exposure.   In one



chromium-alloy plant, four cases  of pulmonary disease with nodular fibrosis and



ventilatory impairment were  reported by Princi et  al. (1962), but no such cases



were found in another similar  plant (Pierce and  Scheel, 1965).



     Bo vet  et  al. (1977)  have suggested  that  workers  exposed to  chromium  in



electroplating operations have  increased frequencies of obstructive respiratory



disease.  The lowest dynamic values of pulmonary function  (e.g., vital ca'pacity,



forced  vital  capacity,  forced expiratory volume, and  forced expiratory flow)



were reported for those workers  displaying the  higher urinary chromium levels.



The  effect  of tobacco  smoke  on  pulmonary function  was minor compared  to the



effect  attributed  to chromium exposure.   Forced expiratory flow  and  forced




expiratory volume were  the pulmonary functions decreased to the greatest extent



as a result of occupational  exposure  to chromium.   The  total number of workers



examined was 44,  and no  data on specific compounds, levels or duration of expo-



sure were reported.
                                     7-134

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      Capodaglio  et  al.  (1975)  demonstrated  alterations  in respiratory function



 among bichromate and chromic acid  production workers.  Observed changes in  chest



 X-ray and  respiratory function parameters were hypothesized to be  the  result  of



 exposure  to chronate,  and  the extent  of  the reduced  pulmonary  function was



 correlated to length of exposure.  Data on the specific compounds  and  levels  of



 exposure were not reported  in  the only available review of this  study  (NIOSH,



 1975).



      Workers  in a  chromite mine and  concentration  plant developed  pulmonary



 markings  (ground-glass  types  1 and  2)  that were attributed  to chromite  dust.



 However, free silica dust  was also present in the air. No clinical  or roentgeno-



 logic evidence of fibrosis was found in the chrcmate workers  (Federal Security



 Agency, 1953).



      Royle (1975)   has  investigated   the   occurrence  of  various  respiratory



 symptoms among British  electroplaters exposed  to chromic acid.  A total of 997



 platers  and  1117 controls completed  a Medical  Questionnaire  on  Respiratory



 Symptoms.  There was a  significant (p<0.025) occurrence of  attacks  of bronchitis



 in platers  (28.2?) compared  to  controls (23.7*).  Platers also  had  significantly



 higher incidences of haemoptysis,  perennial nasal catarrah, and Grade 2 habitual



 winter cough and Grade  2 winter phlegm production. Ratative asthma  was  indicated



 in 13.1? of the platers at a 2.5$ level of  significance.  Nasal and  skin ulcers



 were  significantly  higher in  platers.   Smoking histories were  comparable in



 intensity  and longevity for both  populations.    No clinical testing or medical



 examinations were performed  to  verify the findings of  the Medical Questionnaire.



 Controls were more  frequently  exposed  to a variety  of "dusts" in  present or



 previous employment  situations  or both, and were also exposed to  asbestos  much



more so than was  the study population.   Air samples  taken  between  the  years of



 1969  and  1970 were  <0.03  mg/nr in all  but  two cases (these  were reported as
                                    7-135

-------
<0.1 mg/m ) .  Dust  samples  generally ranged from  0.3 to 97.0mg/g,  but were


reported as high as 298  mg/g.  Members of both the study and control populations


were exposed  to a  variety  of additional  compounds  that could  cause similar


symptoms  (asbestos,  cadmiun,  nickel), a  cu r cum stance that, detracts  from the


overall quality of the study as does the use of medical questionnaires without


any medical examinations to  substantiate  these subjective findings.


     Lucas and Kramkowskl (1975)  have examined the occurrence of abnormal medi-


cal findings in a "hard"  chromium electroplating processing plant.  A total  of 11


workers were screened for various medical complaints  associated with exposure to


chromic acid in electroplating operations.   Average  age  of  the workers was 39


years (range 22  to 54) , with  an average occupational exposure of 7.5 years (range


3  to  16  years).    Average  airborne chromium  (VI)   levels  were reported  as

          •3                                    -3
0.004 mg/m , with a range of  >0.001 to 0.020 mg/m , well within acceptable limits


of the current  NIOSH standards.   Ventilation systems appeared to be functioning


adequately, and  work surfaces adjacent to the  electroplating site were free from


contamination by chromium trioxide, CrO .  A summary of medical findings  is found


in Table 7-25.   Despite the degree  of  protection offered by  ventilation and


protective clothing  and equipment,  workers displayed  a  significant  number of


adverse medical  effects  at presumably "safe" levels of exposure.


     In addition,  a  fine nodular  pneumoconiosis has  been  reported in  a few


chromite miners  in South Africa (Sluis-Cremer and duToit, 1968).  The available


evidence suggests that the pneumoconiosis  is due to deposition of chromite dust


in the lung  tissue,  and that  the condition is  benign  and caused  no fibrosis.


Other components of  the  ore, such as  iron,  may also have been responsible for the


observed fine nodular pneumoconiosis.


     7.2.5.2.   RENAL EFFECTS CF CHROMIUM —Several authors have  reported kidney


damage following the  deliberate ingestion or therapeutic application of chromium
                                    7-136

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                                     TABLE 7-25

       Medical Complaints  of Workers  in  "hard" Chromium Electroplating Plant3
   Frequency                                  Symptom
        1                              Nasal irritation

        U                              Nasal soreness >c

        6                              Runny nose (chronic) 'G

        4                              Frequent nose bleeds

        2                              Ulceration of nasal septum

        9                              Scars indicating previous
                                        ulceration of the nasal  septum

        1                              Perforated nasal  septun

        5                              Chronic coughing episodes

        2                              Pulmonary distress indicative
                                        of emphysema

        7                              Current skin sores

        9                              Scars indicating healed  chrome
                                        ulceration on skin

        5                              Gastric distress

        1                              Chemical diabetes

        1                              Ocular pterygium  on corneal
                                        conjunctiva

        1                              Advanced renal  carcinoma (1967
                                        diagnosis; 1968, one kidney
                                        removed;  1973,  metastasized
                                        to second  kidney; 1971*,  cobalt
                                        treatment
^Source:  Lucas and Kramkowski,  1975                           "     ~~
 Previous to occurrence of  nasal  perforation.
 Two occurences were attributed  to  cold  outdoor  temperatures.
 Three individuals were reported  as moderate to  heavy smokers.
                                      7-137

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compounds (Brieger,  1920;  Godlman  and Karotkin, 1935; Major  1922;  Partington,




1950; Rambousek,  1913).   Twelve persons  died after  the application  of  anti-




scabietic ointment in which sulfur had  been  replaced  with Cr(VI).  Necrosis of



the skin developed at sites of  application and was followed  or  accompanied by




nausea,  vomiting,  shock,  and  coma.    Urinalysis revealed  albumin  and  blood.




Post-mortem findings  included  tabular  necrosis  and  hyperemia  of  the kidneys




(Brieger, 1920).




     Only one of the  above  studies (Goldman and Karotkin,  1935) was available for




review.  It is difficult to determine if chromium has a  direct  toxic effect on




the kidneys or  alters the  normal homeostasis of the body, thereby exerting its




effect.  Evidence to  support the first view comes from a study conduted by Mutti




et al.   (1979).   They  examined welders  and chromium platers,  and found that




workers with a higher degree of exposure to chromium showed a pattern of nephro-



toxicity, as evidenced by increases in the indices for renal tubular  damage.  The




length of  exposure and concentration of  chromium  were  not  specified.   It is




interesting to  note  that  the workers  who had  a higher  degree  of  exposure to




chromium also demonstrated higher urinary chromium  values.




     Pederson and  Mersch  (1978)  reported  the incident of  a woman who ingested




10 mil of a 50% chromic acid solution with 150  mS,  of Coca-Cola.  Vomiting occurred




several times in the hospital,  where gastric lavage was performed and activated




charcoal, magnesia, and milk were administered.  Remedialysis was initiated 18




hours after ingestion.  Blood samples were consistent with anemia, which was most




pronounced by the  eighth day of hospitalization.   Slight granulocytopenia was




observed  on  the third day.  There  was significant proteinuria  (1.9 g)  on the




first day.




     Bilirubin levels rose during the  first 3 days.  One day after ingestion, the




patient was admitted to the renal unit;  the serum chromium was 1.37  ug/mJl (2.9^
                                     7-138

-------
mg Cr/SL whole blood) .  Dialysis treatment was  instituted, which accelerated the




removal  of  chromiun from the  serum.   During the patient's  stay,  no signs of




gastrointestinal  or cerebral  disturbance  were  noted.   Hepatic  function was




normal upon discharge, following a hospital stay of  11 days.








     7.2.5.3.  MIS CELLAN30 US TO HC EFFECTS  —Mancuso (1951) reported  that chro-




mate  workers  frequently  showed  excessive  susceptibility  to  inflammatory and




ulcerative  conditions  of the  gastrointestinal  tract caused by  ingestion of




chromium.




     Hepatic injury, apparently due to exposure to chromic acid mist from plating




baths, has been  reported ( Pascale et  al., 1952).  One woman who had  been employed




for 5 years at a chromium plating factory was hospitalized with jaundice and was




found to be excreting significant amounts  of chromium.  A liver biopsy specimen




showed microscopic  changes  resembling those found in toxic  hepatitis.   Eight




coworkers were screened for urinary  chromiun excretion in an  effort to investi-




gate the possibility that  the hepatic damage was  of occupational origin.  Four of




the workers were found to be excreting significant amounts of chromiun.  In three




workers who had  been exposed  to chromic acid mists  for  1 to 4 years , liver biopsy




specimens and a series of  12  hepatic  function  tests  showed  mild  to moderate




abnormalities.  The correlation of degree of hepatic injury with the concentra-




tion of chromic acid mist and information on controls were  not available.




     Frenkiel and Albert  (1976) reported that  traumatic injury of the tympanic




membrane and external  auditory canal,  and changes  in middle ear  mucosa  were




suffered by   a  worker who  had  fallen  into a vat of  chromic acid.  Extensive




chemical burns were sustained,  along with a 20 decibel conductive hearing loss




stemming from an acid burn to the right tympanic membrane.
                                    7-139

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 7.3   SUMMARY OF TOXIC EFFECTS OTHER THAN CANCER FOLLOWING EXPOSURE TO CHROMIUM



 COMPOUNDS.




      Inhalation exposure is the most predominant route of exposure in industry to




 chromiun compounds and the route most extensively investigated.  Local effects on




 the respiratory system are  the primary toxic effects observed in workers exposed




 to chromium in the atmosphere.  Cr(VI) , in the form  of  chromic  acid,  has been




 associated for  many years   with the  development  of  perforations  of  the nasal




 septun.  The Implication of  chromic acid as  the causative agent results from the




 common  occurrence of  this disorder  in the  chromium-pi at ing industry,  where




 exposure is restricted  to this Cr(VI) compound. Other Cr(VI) compounds may also




 participate in the etiology  of perforated nasal septuns, since this disorder has




 been  reported  in  the chronate  manufacturing  industry,  where the predominate




 exposures are to  Cr(III) and  the  Cr(VI)  compounds, sodium chromate and sodium




 dichrcmate; however,  chromic acid  mist may  also be  present in these plants.   It




 is interesting to  note that  nasal  septun perforation has not been reported as an




 occupational  hazard in the chrcme leather tanning industry or the chrcme pignent




 industry, both of which  exclusively use  Cr(VI) , although  these  industries  are




 associated with severe chromium dermatitis.  The lack of perforated nasal septums




 in these industries may result from differences in  the physical or chemical form




 of the chromiun, droplets in the  tanning industry and participates in the pignent




 industry as compared  to  the  chromic acid mist generated in the plating industry.




The measurements  of  chromic acid associated  with  perforated  septums in  the




 chrome-pi at ing industry is  X).1 mg/m3 (see Table 7-21); however, it is not known




 if lower  concentrations  are also  effective.   Also,  severe  irritation  of  the




 throat and lower respiratory tract have been associated with chromiun  compounds




 at concentrations  as low as  0.12 mg/m3. • Again, as with perforated nasal septum,




this respiratory tract irritation is primarily associated with Cr(VI) .   Hyper-
                                    7-1^0

-------
 sensitivity  may result from dermal or inhalation exposure to either Cr(VI) or



 Cr(III); however, there is little information available on the levels of exposure



 necessary to induce an allergic response.



      Little  information  is available  on  systemic  effects  of  inhalation of



 chromium  compounds,  although  Pascale  et al.  (1952)  and Mutti  et al.   (1979)



 reported liver  injury in a chromate worker and  kidney injury in a welder exposed



 to  chromium, respectively.  Acute exposure of animals  using a variety of  routes



 of  administration  (Section  7.1.2)  have  indicated that both Cr(VI) and Cr(III)



 compounds can produce kidney and liver danage ,  although the dose levels employed



 were  relatively higi.  From the evidence available from  both  human  case reports



 and animals  studies,  it can  only be speculated whether the  kidneys  and liver may



 be  target organs following chronic exposure  to chromium  compounds.



      Although inhalation studies of occupational exposure to chromium indicate



 that  exposure to some chromium compounds can result in perforation  of the nasal



 septum, irritation of the respiratory tract,  pne uno coni osis, bronchitis, chronic



 lung  congestion, and possible  liver and kidney damage (as supported by target



 organ toxicity in acute animal studies) ,  there are insufficient data available to



 make  a quantitative risk assessment  for  either  chromium as a class  or individual



 chromiun compounds from these inhalation studies.  The only studies  that provide



 any exposure data  (and this  is  of  limited and  questionable  quality,  since



 measurements were not made contemporary  with exposure and  personal  habits, such



 as  picking of the nose, may  result in high local concentrations of  chromiun) are



 the studies  of  the occurrence  of  perforated nasal sept urns.  Since perforated



 nasal septun  results from the  local destruction of the  mucous  membrane  of the



 nose, this  does  not represent a systemic  effect of chromium.  Also, in the study



 by Nettesheim et al.  (1971), inhalation exposure of mice to chromiun resulted in



marked effects to the respiratory tract.  These effects,  including hyperplasia
                                    7-TH

-------
and necrosis, were likely to have resulted from the  severe local irritation of



the  cells  lining the  air ways.   It would not  be  expected that  exposure  to



chromium by any other route would result in this  disorder,  and  although these



data can be  used to derive an  acceptable  inhalation  exposure level,  they are



inadequate for determining safe levels of chromiun by  all  routes  of exposure.



The limited information on other systemic effects of inhaling chromium, liver and



kidney damage, contains both insufficient exposure data and too few case reports



to form a firm association  between exposure and effect.



     There  are only  a few instances of human exposure to overtly toxic levels of



chromium compounds by ingestion, and these represent  acute exposure to massive



doses which provide little  information on the safe levels of chromium following



chronic exposure.  A number of animal studies  have been performed in which the



chromium compound was  administered in the food, water, or by gavage.  The acute



oral toxicity data indicate that Cr(VI) is approximately 2 or 3 orders of magni-



tude more toxic than Cr( III) , with the latter toxic at the level  of g/kg body



weight  (Section 7.1.2).  The  difference  in valence  state  may  be less relevant



following chronic  or subchronic ingestion  of chromiun,  since it is suggested that



Cr(VI)   is  reduced to  Cr(III)  under the  acid  conditions  of the stomach.   The



determination as to whether Cr(III)  or Cr(VI) is more toxic after chronic expo-



sure, however, cannot  be made, since none of the studies employed a sufficiently



high dose to produce a  toxic  effect.



     The only study in which  an  effect  was observed  was  that  of Ivankovic and




Preussman (1975) in which rats were  fed diets containing 2 or 5% CrpO-tCr"1"-*),  5



days/week for 90 days.  The only observed  effect was a reduction in  the weight of



the liver and spleen in the  treated male rats as compared with liver and spleen



weights of  control animals.   Similar results were observed in female rats main-



tained on the same diet.  Neither organ showed macroscopic or mi ores oo pic abnor-
                                    7-142

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 malities, and the authors concluded that these changes  were not toxicologically



 important.  In a larger 2-year study using the same experimental  procedure and 60



 animals of each sex per  group,  Ivankovic and Preussman (1975) did not mention any



 treatment-related changes  in organ weight, although it  was  mentioned  that  no



 signs  of  chronic  toxicity were observed.   Therefore, it  is unclear whether the



 slight change in organ weight observed in the small nunber of  animals in the 90-



 day study was the result of spurious observation due to  the small  group size, or



 whether the  5% exposure level  represents  a  true NOAEL  (no-observed-adverse-



 effect-level).  No matter whether this 90-day study represents a NQftEL  or an NCEL



 (no-observed-effect-level) , the small  group size makes  this study very tenuous



 as the basis for quantitative risk assessment.



     In the absence of any data on  effect levels following chronic exposure  to



 chromium,  the  U.S.   EPA,  in the  Ambient  Water Quality  Criteria Document  for



 Chromium  (U.S.  EPA,  1980a),  derived acceptably daily  intake values  (ADIs)  of



 0.175 and  125 mg/day/man for Cr(VI)  and  Cr(III), respectively.  These ADIs were




 derived by using the  highest  NCEL available for each valence state.  For Cr(VI),



 the study  of  MaoKenzie  et  al.   (1958) was  used, in which rats  were  exposed  to



 several levels of chromium in the form  of KpCrOj. up to 25 ppm  in the drinking



 water for 1 year, while for Cr(III), the  chronic study of Ivankovic and Preussman



 (1975) was used, in which rats were fed diets containing up  to 5% Cr20_ for  2



 years.   The  ADIs for  both Cr(VI) and  Cr(III)  were expressed  as mg/d of  the



 compound administered.  Although these ADIs were, derived, it is not apparent  from



 the toxi col ogle information  available whether  the ADIs  are more  appropriate for



the specific chromiun compounds tested, KpCrOj. and Cr_0_, rather than the general



 classes of Cr(VI)  and Cr(III) .  This is particularly hard  to determine, since no



toxic  effects  were observed in these chronic animals studies.

-------
                     8.  CURRENT REGULATIONS AND STANDARDS




     A number of recommended standards presently exist for permissible levels of




chromium in both air and water.  The National Academy of Science (NAS, 1980) and




the U.S. Environmental Protection Agency (U.S. EPA, 1980a), in regard to ambient




water quality criteria, have stated that  a distinction  between the Cr(III) and




(VI) forms should  be  made when considering the formulation  of regulations and




exposure criteria.








8.1.  OCCUPATIONAL EXPOSURE




     NIOSH (1973,  1975), OSHA (1978),  and ACGIH (1981)  have recommended various




exposure limits for chromium.  These values are  based on the chemical form of the




chromium compounds or their solubilities.   Table 8-1 outlines the various United




States occupational standards for chromium compounds.








8.2.  EXPOSURE TO CHROMIUM IN AMBIENT  WATER.




     A number of standards exist for chromium in ambient water or drinking water.




In general,  the standards for occupational exposure to chromium in the air allow




for a greater uptake  of chromium  than  by the uptake of chromium  from  drinking




water.  The  U.S.  EPA (1980a) has  estimated the daily intake  of  chromium from



drinking water to be 5 ng/day.  Table  8-2 outlines  various recommended  or esta-




blished standards for chromium in the  United States.




     The U.S. EPA  (1980a) recently  proposed several ambient  water quality cri-




teria for  chromium.   Based on methodology  outlined  in the Federal Register




(M5 FR 79353),  acceptable  daily  intakes   (ADIs),  no-observable-adverse-effect




levels (NOAELs), and bioconcentration  factors  (BCFs) obtained from experimental




animal studies,  separate water  quality  criteria were  proposed.   Table 8-3
                                      8-1

-------
                                     TABLE  8-1

          Recommended Occupational  Standards  and Recommended Criteria for
                      Chromium Compounds  in the United States
Chemical Form
Non- carcinogenic chromium (VI )a
Carcinogenic chromium (VI)
Chromic acid (as chromium trioxide)
Soluble chromic or chromous salt
Insoluble salts or chromium metal
Chromium metal
Chromium (II) compounds
Chromium (III) compounds
Chromium (IV) compounds
water soluble
water insoluble
Chromite ore
Chromium: soluble chromic and
chromous salts
Standard (^g/m^)
25 TWA
50 Maximum
1
50 TWA
100 Maximum
500 TWA
1000
500 TWA
500 TWA
500 TWA
50 TWA
50 TWA
50
500 TWA
Reference
NIOSH, 1975
NIOSH, 1975
NIOSH, 1973
OSHA, 1978
OSHA, 1978
ACGIH, 1981°
ACGIH, 1981
ACGIH, 1981
ACGIH, 1981
ACGIH, 1981
ACGIH, 1981
ACGIH, 1981
wIOSH listed "non-carcinogenic" chromium VI compounds as the monochromates and
dichromates (bichromates) of:   hydrogen, lithium,  potassium, rubidium, cesium,
ammonium, and chromic oxide (chromic acid anhydride).

NIOSH listed "carcinogenic" chromium VI compounds  as all other chromium compounds not
included in the "non-carcinogenic" chromium VI listed above.

These are also known as the TLV values established by ACGIH.

ACGIH listed these classes of chromium compounds as substances associated with
industrial use that have been recognized as carcinogens.
                                         8-2

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                                      TABLE  8-2

                         Recommended Standards for Chromium
                       in Ambient Waters in the United States
Chemical Form
Chromium (VI)
Total Chromium
Medium
drinking water
total
domestic water
Criteria (\Lg/H)
50
50
Reference

U.S. Public Health
Service (USPHS), 1962
U.S. EPA, 1976
Total Chromium


Chromium (VI)
Chromium
supply

freshwater
(aquatic life)

livestock water
community water
systems and non-
community water
systems
 100
1000
  50
U.S. EPA, 1976
National Academy of
Science and National
Academy of Engineering
(NAS/NAE), 1972

40 CFR 141,11
                                          8-3

-------
                                                     TABLE  8-3
                                        Ambient Water Quality Criteria for
                                          the Protection of Human Health3
Chemical NOAEL
Form (mg/£)
Chromium (III)b 50,000
i
"^ Chromium (VI) 25
Rat
NOAEL
(mg/d/kg)
1786
2.50
ADI
for man
(mg/d/man) BCF
125 16
0.175 16
Calculated
Criteria (|ig/&)
59,000
83
aSource: U.S. EPA, 1980a
 Revised ADI and criterion.  Published values  (45  FR 79331)  were incorrect.   If exposure to trivalent chromium
results  only from the eating of fish and shellfish, then the calculated ambient water criterion for chromium
(III) is proposed  as 1200 mg/Jl (1.2 x 10

-------
summarizes the proposed U.S.  EPA (1980a)  ambient water quality criteria for the




protection of human health.



     The U.S. EPA (1980a) also has proposed several ambient  water  quality cri-



teria for the  protection  of  aquatic life.   Table 8-4 summarizes  the  proposed



criteria for the protection of aquatic life.








8.3. EXPOSURE TO CHROMIUM IN  AMBIENT  AIR.



     No federal or  state  (other  than the state of Maine,  i.e.,  0.05 ng/nr for



mean annual and 0.3 \ig/fn  for mean daily limits) ambient air  chromium standards



have been proposed.  No United States emission standards for chromium were found




in the available literature.  TheU.S.S.R. recommends a "sanitary clearance zone"



of  1000 m for  plants  discharging 200 kg Cr(VI) per day, and 2000  m for plants



discharging 1000 kg per day (NAS, 197*0.
                                      8-5

-------
                                    TABLE  8-4

                    Calculated Ambient Water Quality Criteria
                       for the Protection  of Aquatic Life*
Chemical
Form
Freshwater Life
24-hour Average Maximum
(Hg/fc) (|ig/fc)
Marine Life
24-hour Average
(|ig/A)
Maximum
(fig/A)
Chromium (III)
Chromium (VI)
    44
(chronic value
   toxicity)

     0.29
NR
21
10,300 (acute
toxicity value)
    18
 NR
1260
•Source:U.S. EPA, 1980a
NR = Not recorded
                                           8-6

-------
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   U.S. Environmental Protection Agency '
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   230 South  Deaibc-n  Slreet
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