United Slates          .
EnviionmenMl PioU'ciioij
Ayenr.y
Office of Hi.-alih •ind
Environmental Assassin en I
Wasluniiion DC ^0460
EPA 600 8-82-008I-"
August 1983
Final Report
                                                             FINAL

                                                             RliPORT

                                                           PB84-100056

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                        NOTICl:

This  document has  been reviewed in ncconiuiK c wi.;h
U.S..  Lnv i ronme.ntai  I'rot.cction  Agency  polic;,' ;iniJ
approvi'J i'or publication.   '-lent i <>n of t r;nK: runncs
or commercial products Jui'S not riHiSt i. tut '-• en Ju r ••;(_•-
nont  or- recoiniiieiiuia t. ion for'use.

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                                   EPA-600/8-82-008F
                                         August 1983
                                        Pinal Report
Document for Toluene
             FINAL

             RLTORT
  US ENVIRONMENTAL PROTECTION AGENCY
     Office of Research and Development
 Office of Health .-ind Environmental Assessment
  Environmental Criteria and Assessment Office
      Research Tnangie Park. NC 27711

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                                    PREFACE
     This document has been  prepared  by  the  Environmental  Criteria  and  Assess-
ment OfTice of the U.S. Environmental Protection Agency (EPA).  The document  was
originally  developed  to  support  U.S. EPA  decision-making regarding  possible
regulation of toluene as a hazardous air pollutant.  The scope  of the  document
has  since  been  expanded  to  address  multimedia aspects and  thus  enables  the
document to serve as a "source document" for  other U.S. EPA  programs requiring
comprehensive information concerning the health effects of  toluene.
     The Health Assessment Document for Toluene was reviewed and critiqued by  the
Environmental Health Committee of the U.S.  EPA Science Advisory Board in August
1982.  This committee provides advice on scientific matters  to the Administrator
of the U.S. Environmental Protection Agency.
     In  the  development  of  the  assessment  document,  the scientific literature
has been critically evaluated and  the conclusions presented  in such a manner that
the toxicity  of toluene  and related characteristics  are qualitatively  identi-
fied.  Observed effect levels and ether measures of dose-response relationships
are discussed, where appropriate,  in order that the nature of the adverse health
responses are placed in perspective with observed environmental  levels.
                                      ii

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                                   ABSTRACT
     Toluene  is  the  most  prevalent  hydrocarbon  in  the  atmosphere.    Levels
generally range from  0.11-57 ppb.  Levels  in water generally are below 10 ppb.
Gasoline usage and automobile exhaust represent the largest atmospheric source.
Ov4r 3 million metric  tons of toluene  are produced annually in the  United States.
     ^"ailable evidence  associated  with effects  upon humans and  experimental
animals indicates that the health effect  of primary concern is dysfunction of the
central nervous  system (CNS).    However,  observed effects are associated with
exposure levels greatly in excess of those levels in the environment.   Dysfunc-
tion of the CNS may occur during short-term (£8 hours) exposure  tn 100-300 ppm.
     Toluene  has not demonstrated any overt signs  of kidney or liver damage upon
animal experimentation.  It was non-carcinogenic in rats exposed  to 300 ppm for
24  months.    However,  the full  extent  of  toluene's  carcinogenic  potential  is
currently being evaluated, at higher exposure levels,  in a lifetime bioassay of
rodents  in   the  National  Toxicology  Program.    Toluene  is   classified  as
provisionally non-mutagenic, and its teratogenic potential has   not been  fully
explored.
     The results  of the available evidence indicate  that  exposure  to  environ-
mental levels of toluene  is  unlikely to  constitute  a  significant hazard to the
general population.
                                      ill

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                               TABLE OF CONTENTS
LIST OF TABLES                                                                xi

LIST OF FIGURES                                                               xv

AUTHORS, CONTRIBUTORS,  AND  REVIEWERS                                         xvi

SCIENCE-ADVISORY BOARD  ENVIRONMENTAL  HEALTH  COMMITTEE                        xix

1.  EXECUTIVE SUMMARY                                                        1-1

    1.1.   ENVIRONMENTAL  SOURCES,  FATE,  AND  LEVELS                           1-1
    1.2.   EFFECTS ON HUMANS                 '                                1-3
    1.3.   ANIMAL STUDIES                                                   1-5
    1.4.   ABsohPTioN,  iJi.-yr.-uBijTiON,  METABOLISM,  ELIMINATION,
           AND  RELATED  PHAiiMACOKI.NETICS                                      1-6
    1.5.   CARCINOGENICITY, MUTAGENICITY,  AND  TERATOGENIC1TY                 1-6
    1.6.   EFFECTS ON ECOSYSTEMS                                             1-7
    1.7.   HEALTH EFFECTS SUMMARY                                            1-8
    1.8.   RESEARCH  NEEDS                                                   1-9

2.  INTRODUCTION                                                             2-1

3.  PHYSICAL  AND CHEMICAL PROPERTIES                                         3-1

    3.1,   SYNONYMS  AND TRADE  NAMES                                          3-1
    3-2.   IDENTIFICATION NUMBERS                                            3-1
    3-3-   STRUCTURE, MOLECULAR FORMULA, AND MOLECULAR WEIGHT                3-1
    3.4.   PHYSICAL  F'R&PFRTIES                                              3-1

           3.4.1.    Description                                              3-1
           3.4.1*.    Other Physical Properties                                3-1
           3-^.3-    Sigr.if icance of Physical Properties with
                     Rucpect to Environmental Behavior                        3-2

    .3.5.   CHEMICAL  HhOPEff] ii,S                                              3-3
    3.6.   REFERENCES                                                        3-5

14,  PRODUCTION', USE, AND  RELEASES  TO  THE ENVIRONMENT                         i)-i

    4.1.   MANIiTACTChlNG  PRDCEHS TECHNOLOGY                                  4-1.

           U. 1. I.    y^tfoleum  Refining Processes                             '(-1

                     A. I, 1.1.   CATALYTIC  REFORMING                           4-1
                     H.i.1.2.   PYHOLYTIC  CRACKING                             4-1

           4.1.2.    By-Product of  Styrene Production                         4-3
           4.1.J}.    By-Product of  Coke-Oven  Operation                        4-3

    4.2.   PROD'jCEK."                                                         4-3
    4.3.   USERS                                                            4-10
    4.4.   ENVIRONMENTAL  RELEASE                                            4-13

                                       iv

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                           TABLE OF CONTENTS (cont.)
           I.U.I.   Emission from Production Sources                       4-13
           4.4.2.   Emission from Toluene Usage                            4-18
           4.4.3.   Emission from Inadvertent Sources                      4-23
           4.4.4,   Non-anthropogenic Sources                              4-23
           4.4.5.   Sum of Emissions from All Sources                      4-26

    4.5.   USE OF TOLUENE IN CONSUMER PRODUCTS                             4-25
    4.6.   REFERENCES                                                      4-29

5.  ABATEMENT PRACTICES IN INDUSTRY                                         5-1

    5.1.   ABATEMENT PRACTICES FOR INADVERTENT SOURCES                      5-1
    5.2.   ABATEMENT PRACTICES FOR SOLVENT USAGE                            5-2
    5.3.   ABATEMENT FOR COKE OVEN EMISSIONS                                5-2
    5.4.   ABATEMENT FOR EMISSIONS FROM MANUFACTURING SITES                 5-2
    5.5.   ABATEMENT PRACTICES FOR RAW AND FINISHED WATERS                  5-3
    5.6.   ECONOMIC BENEFITS OF CONTROLLING TOLUENE EMISSIONS               5-3
    5.7-   REFERENCES                                                       5-3

6.  ENVIRONMENTAL FATE, TRANSPORT, AND PERSISTENCE                          6-1

    6.1.   AIR                                                              6-1

           6.-1.1.   Fate in Air                                             6-1
           6.1.2.   Transport                                               6-5

    6.2.   AQUATIC MEDIA                                                    6-5

           6.2.1.   Fate                                                    6-5
           6.2.2.   Transport                                               6-6

    6.3.   SOIL                                                             6-7

           6.3.1.   Fate                                                    6-7
           6.3.2.   Transport                                               6-6

                    6.3-2.1.  SOIL TO AIR                                   6-8

    6.4.   ENVIRONMENTAL PERSISTENCE                                        6-8

           6.4.1.   Biodegradation and Biotransformation                    6-8


                    6.4.1.1.  MIXED CULTURES                                6-8
                    6.4.1.2.  PURE CULTURES                                6-10

    6.5.   REFERENCES                                                      6-13

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                           TABLE OF CONTENTS (cont.)

                                                                           Page

7.  ENVIRONMENTAL AND OCCUPATIONAL CONCENTRATIONS                           7-1

    7.1.   ENVIRONMENTAL LEVELS                                             7-1

           7.1.1.   Air                                                     7-1
           7.1.2.   Aqueous Media                                           7-9

                    7-1.2.1.  SURFACE WATERS                                7-9
                    7-1.2.2.  INDUSTRIAL WASTEWATERS                        7-11
                    7. 1.2.3.  PUBLICLY-OWED TREATMENT WORKS (POTW)         7-11
                    7.1.2.1.  UNDERGROUND WATER                            7-11
                    7.1-2.5.  DRINKING WATER                               7-11
                    7.1.2.6.  RAINWATER                                    7-15

           7.1.3-   Sediment                                               7-15
           7.1.1.   Edible Aquatic Organisms                               7-15
           7.1.5.   Solia Wastes and Leachates                             7-15

    7.2.   OCCUPATIONAL CONCENTRATIONS                                     7-15
    7.3.   CIGARETTE SMOKE                                                 7-20
    7.1.   REFERENCES                                                      7-20

8.  ANALYTICAL METHODOLOGY    .                                              8-1

    8.1.   AIR                                                              8-1

           8.1.1.   Ambient Air                                             8-1

                    8.1.1.1.  SAMPLING                                      8-1
                    8.1.1.2.  ANALYSIS                                      8-2
                    8.1.1.3.  PREFERRED METHOD                              8-3
                    8.1.1.1.  DETECTION LIMITS                              8-1

           8.1.2.   Occupational Air                                        8-1

                    6.1.2.1.  SAMPLING                                      8-1
                    8.1.2.2.  ANALYSIS                                      8-5
                    8.1.2.3.  PREFERRED METHOD                              8-6
                    8.1.2.1.  DETECTION LIMIT                               6-7

           8.1.3.   Forensic Air                                            8-7
           8.1.1.   Gaseous Products from Pyrolysis of Organic Wastes       8-7

    8.2.   WATER                                                            8-7

           8.2.1.   Sampling                                                8-7
           8.2.2.   Analysis                                                8-8

                    8.2.2.1.  PURGE AND TRAP                                8-8
                    8.2.2.2.  HKADSPACE ANALYSIS                            8-9
                    8.2.2,3.  SORPTION ON SOLID SORBENTS                   8-10
                                       vi

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                           TABLE  OF  CONTENTS  (cent.)

                                                                           Page

    8.3.    SOILS AND SEDIMENTS                    ,                          8-10

           8.3.1.   Sampling                                               8-10
           8.3.2.   Analysis                                               8-11

    8.U.    CRUDE OIL AND ORGANIC  SOLVENTS                                   8-11
    8.5.    BIOLOGICAL SAMPLES                                              8-12

           8.5.1.   Blood                                                  8-12
           8.5.2.   Urine                                                  8-12
           8.5.3.   Mother's Milk                                          8-13

    8.6.    FOODS                                                           8-13
    8.7.    CIGARETTE SMOKE                                                 8-13
    8.8.    REFERENCES                                                      8-14

9.  EXPOSED POPULATIONS                                                    9-1

    9.1.    REFERENCES                                                      9-3

10. ESTIMATE OF HUMAN EXPOSURE                                             10-1

    10.1.  EXPOSURE VIA INHALATION                                         10-2
    10.2.  INGESTION EXPOSURE BASED ON MONITORING DATA                      10-9

           10.2.1.  Exposure from Drinking Water                           10-9
           10.2.2.  Exposure from Edible Aquatic Organisms                 10-9

    10.3.  OCCUPATIONAL EXPOSURE                                            10-9
    10.4.  CIGARETTE SMOKERS                                              10-10
    10.5.  LIMITATIONS OF EXPOSURE ESTIMATE BASED ON MONITORING DATA       10-10
    10.6.  COMPARISON BETWEEN EXPOSURE DATA BASED ON THEORETICAL AND
           EXPERIMENTAL VALUES                                            10-11
    10.7.  REFERENCES                                                     10-12

11. EFFECTS ON HUMANS                                                      11-1

    11.1.  EFFECTS ON THE NERVOUS SYSTEM                                   11-1

           11.1.1.  Central Nervous System                                 11-1

                    11.1.1.1. ACUTE EFFECTS                                11-1
                    11.1.1.2. SUBCHHONIC AND CHRONIC EFFECTS               11-9

           11.1.2.  Peripheral  Nervous System                             11-16

    11.2.  EFFECTS ON THE BLOOD AND HEMATOPOIETIC TISSUE                  11-22

           11.2.1.  Bone Marrow                                           11-22
           11.2.2.  Blood Coagulation                                     11-29
           11.2.3.  Phagocytic Activity of Leukocytes                     11-30
           11.2.4.  Immunocompetencs                                      11-30

                                      vii

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                           TABLE  OF  CONTENTS  (cont.)
    11.3.   EFFECTS  ON- THE  LIVER                                            11-30
    11.1.   EFFECTS  ON THE  KIDNEYS                                          11-31
    11.5.   EFFECTS  ON THE  HEART                                            11-38
    11.6.   EFFECTS  ON MENSTRUATION                                         11-39
    11.7.   EFFECTS  ON THE  RESPIRATORY TRACT  AND THE  EYES                   11-10

           11.7-1.   Effects of Exposure                                   11-140

    11.8.   EFFECTS  ON THE  SKIN                                            11-12
    11.9.   SUMMARY                                                         11-12
    11.10. REFERENCES                                                     11-16

12.  ANIMAL TOXICOLOGY                                                      12-1

    12.1.   SPECIES  SENSITIVITY                                             12-1

           12.1.1.   Acute  Exposure  to Toluene                              12-1

                    12.1.1.1.  ACUTE INHALATION                             12-1
                    12.1.1.2.  ACUTE ORAL TOXICITY                         12-17
                    12.1.1.3.  ACUTE EFFECTS  FROM INTRAPERITONEAL
                              INJECTION                                   12-17
                    12.1.1.1.  ACUTE EFFECTS  FROM SUBCUTANEOUS INACTION    12-18
                    12.1.1.5.  ACUTE EFFECTS  FROM INTRAVENOUS INJECTION     12-18
                    12.1.1.6.  ACUTE AND SUBACUTE EFFECTS  OF PERCUTANEOUS
                              APPLICATION                                 12-13

           12.1.2.   Subchronic and  Chronic Exposure to Toluene            12-18

    12.2.   EFFECTS  ON LIVER, KIDNEY, AND LUNGS                            12-22

           12.2.1.   Liver                                                  12-21
           12.2.2.   Kidney                                                12-27
           12.2.3-   .Lungs                                                  12-27

    12.3.   BEHAVIORAL TOXICITY AND  CENTRAL NEHVOUS SYSTEM EFFECTS         12-28

           12.3.1.   Effect of Solvent-Sniffing Abuse                      12-29
           12.3-2.   Effects on Simple and Complex Behavioral Performance   12-32
           12.3.3.   Effect on Electrical Activity of the  Brain and Sleep   12-38
           12.3.1.   Effect on Neuromcdulators                             12-11
           12.3-5.   Minimal Effect  Levels                                 12-11

    12.1.   EFFECTS  ON OTHER ORGANS                                         12-12

           12.1.1.   Blood-Forming Organs                                  12-12
           12.1.2.   Cardiovascular  Effects                                12-16
           12.1.3.   Gonadal Effects                                       12-16

    12.5.   SUMMARY                                                         12-1?
    12.6.   REFERENCES                                                     12-50

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                           TAELE OF CONTENTS (cont.)

                                                                           Page

13.  PHARMACOKINETIC CONSIDERATIONS IN HUMANS AND IN ANIMALS                13-1

    13-1.  ROUTES OF EXPOSURE AND ABSORPTION                               13-1
    13.2.  DISTRIBUTION                                                    13-9
    13-3.  METABOLISM                                                     13-14
    13.4.  EXCRETION                                                      13-19
    13.5.  SUMMARY                                                        13-28
    13.6.  REFERENCES                                                     13-30

14.  CARCINOOENICITY, MUTAGENICITY, AND TERATOGENICITY                      14-1

    14.1.  CARCINOGENICITY                                                 14-1
    14.2.  MUTAGENICITY                                                    14-2

           14.2.1.  Growth Inhibition Tests in Bacteria                    14-2
           14.2.2.  Tests for Gene Mutatjons                               14-4

                    14.2.2.1. ASSAYS USING BACTERIA AND YEAST              14-4
                    14.2.2.2, TK MUTATION IN L5178Y MOUSE
                              LYMPHOMA CELLS                               14-6

           14.2.3.  Tests for Chromosomal Mutations                        14-6

                    14.2,3.1. MICRONUCLEUS TEST IN MICE                    14-6
                    14.2.3-2. MOUSE DOMINANT LETHAL ASSAY                  14-6
                    14.2.3-3. CHROMOSOME ABERRATION STUDIES                14-7
                    14.2.3-4. SISTER CHROMATID EXCHANGE                   14-14

    14.3.  TERATOGENICITY                                                 14-16

           14.3.1.  Animal Studies                                        14-16

    14.4.  SUMMARY                                                        14-25
    14.5.  REFERENCES                                                     14-26

15. SYNERGISMS AND ANTAGONISMS AT THE PHYSIOLOGICAL LEVEL                  15-1

    15.1.  BENZENE AND TOLUENE                                             15-1
    15.2.  XYLENES AND TOLUENE                                             15-2
    15.3.  TOLUENE AND OTHER SOLVENTS                                      15-3
    15.4.  REFERENCES                                                      15-4

16. ECOSYSTEM CONSIDERATIONS                                               16-1

    16.1.  EFFECTS ON VEGETATION                                           16-1

           16.1.1.  Introduction                                           16-1
           16.1.2.  Effects of Toluene on Plants                           16-1

                    16.1.2.1. ALGAE                                        16-1

                              16.1.2.1.1.  Closed System Studies           16-1
                              16.1.2.2.2.  Open Studies                    16-4
                                       ix

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                             I

                           TABLE OF CONTENTS  (cont.)

                                                                           Page

                    16.1.2.2.  EFFECTS ON  HIGHER  PLANTS                      16-4

    16.2.   BIOCONCENTRATION,  BIOACCUMULATION,  AND  BIOMAGNIFICATION
           POTENTIAL                                                       16-7
    16.3.   EFFECTS ON MICROORGANISMS                                      16-14
    16.4.   REFERENCES                                                     16-17

17.  EFFECTS ON AQUATIC SPECIES                                             17-1

    17-1.   GUIDELINES FOR EVALUATION                                       17-1
    17.2.   EFFECTS OF ACCIDENTAL SPILLS                                    17-2
    17.3-   LABORATORY STUDIES OF TOXICITY                                  17-3

           17.3.1.  Lethal Effects                                         17-3

                    17.3.1.1.  FRESHWATER  FISH                               17-3
                    17.3.1.2.  MARINE FISH                                 17-13
                    17.3-1-3.  FRESHWATER  INVERTEBRATES                     17-15
                    17.3-1.4.  MARINE INVERTEBRATES                        17-15

           17.3.2.  Sublethal Effects                                     17-18

                    17.3-2.1.  FISH                                        17-18
                    17.3.2.2.  INVERTEBRATES                               17-22

    17.4.   REFERENCES                                                     17-25

18.  HEALTH EFFECTS SUMMARY                                                 18-1

    18.1.   EXISTING GUIDELINES AND STANDARDS                                18-1

           18.1.1.  Air                                                    18-1
           18.1.2.  Water                                                  18-2
           18.1.3.  Food                                                   18-2

    18.2.   INHALATION EXPOSURES                                            18-3

           18.2.1.  Effects of Single Exposures                             18-3
           18.2.2.  Effects of Intermittent Exposures Over
                    Prolonged Periods                                      18-6

    18.3.   ORAL EXPOSURES                                                  18-9
    18.4.   DERMAL EXPOSURES                                                18-9
    18.5-   RESPONSES OF SPECIAL CONCERN                                   18-10

           18.5.1.  Carcinogenicity                                       18-10
           18.5.2.  Mutagenicity                                          18-10
           18.5.3.  Teratogenicity                                        18-11

    18.6.   REFERENCES                                                     18-13

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                                LIST OF TABLES

Number                                                                   Page

 4^1        U.S. Production of Isolated Toluene in 1978                   4-2

 4-2        Isolated and 'cion-Isolated Toluene Available in the
              United States in 1978                                       4-5

 4-3        Producers of Isolated Toluene from Catalytic Reforming
              in 1978                                                     4-6
 4-4        Producers of Isolated Toluene from Pyrolysis Gasoline

 4-5        Producers of Isolated Toluene from Styrene By-Product         4-9

 4-6        Producers of Isolated Toluene from Coke-Oven Crude
              Light Oils                                                  ^-10

 4-7        Consumption of Isolated and Non-Isolated Toluene in
              Different Usages                                            4-12

 4-8        Consumers of Toluene for the Manufacture of Benzene by
              HDA Process                                                 4- '.3

 4-9        Producers of Toluene Diisocyanat.e (TDI) in 1978               4-15

 4-10       Other Toluene Chemical Intermediate Users in 1978             4-16

 4-11       Toluene Air Emission Factors from Production Sources          4-18

 4-12       Estimated Atmospheric Toluene Emissions from Four Major
              Production Sources                            .              4-19

 4-13       Toluene Emission Factors in Wastewater from Coke Oven
              Operation                      •                             4-21

 4-14       Toluene Released in Different Media from Coke-Oven
              Wastewater                                                  4-22

 4-15       Toluene Emission Factors for Its Uses                         4-23

 4-16       Estimated Toluene Emission from Different Uses                4-24

 4-17       Toluene Released in Aqueous Media from Use as a Solvent
              in Various Industries                                       4-26

 4-18       Toluene Emission from Different Inadvertent Sources           4-27

 4-19       Tc*-al Yearly Release of Toluene into Different Media          4-29

 4-20       Consumer Product Formulations Containing Toluene              4.31
                                       xi

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                            LIST  OF  TABLES  (cont.)

Number

 6-1        Rate Constants for Reactions of Toluene with Reactive
              Species in the Atmosphere                                   6-3

 7-1        Atmospheric Concentrations of Toluene                         7-2

 7-2        Distribution of U.S. Surface Waters Within a Certain
              Toluene Concentration Range                                 7-6

 7-3        Percent Distribution of U.S. Wastewaters Within a
              Certain Toluene Concentration Range                         7-7

 7-4        Detection Frequency of Toluene in Industrial Wastewaters      7-9

 7-5        Toluene Concentrations in Different Work Areas of a
              Rotogravure Plant in Milan, Italy                           7-15

 7-6        Toluene Exposure Levels for Different Occupational Groups     7-16

 7-7        Toluene Concentrations in Work Areas of Leather
              Finishing and Rubber Coating Plants                         7-17

 7-8        Toluene Concentrations in Selected Work Areas of Tire
              Manufacturing Plr\nts                                        7-19

 9-1        Population Distribution and Inhalation Exposure Levels
              of Toluene from Different: Sources                           9-2

 10-1        Concentration of Toluene  (mg/nr) at Different
              Distances (m) From A Sources Emitting 200 Million
              kg/Year Toluene                                            10-5

 10-2        Population Distribution and Inhalation Exposure Levels
              of Toluene from Different Sources                          10-7

 10-3        Toluene Exposure Under Different Exposure Scenarios          10-10

 10-4        Exposed Population and Exposed Amount of Toluene From
              Dispersion Modelling                                       10-15

 11-1        Effects of Controlled 8  Hour Exposures to Pure Toluene
              on Three Human Subjects                                    11-3

 11-2        Effect of Toluene Exposure on the Performance of
              Perceptual Speed and Reaction Time Tests                   11-6

 11-3        Mean Concentrations of Organic Solvents in the Breathing
              Zone of 40 Car Painters                                    11-15
                                      xii

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                            LIST OF  TABLES  (cont.)

Number

11-4        Performance Tests:  Means, Standard Deviations, and
              Significances Between the Group Means (Age-Matched)
              Groups                                                     11-16

11-5        Rorschach Personality Test Variables:  Means, Standard
              Deviations, and Significances Between the Group
              Means (Age-Matched Groups)                                 11-17

11-6        Encephalopathic effects of Chronic Toluene Abuse             11-20

11-7        Results of Neurological.and Muscular Function Tests
              of Toluene-Exposed Female Shoemakers                       11-23

11-8        Results of Blood Examinations Performed on Toluene-
              Exposed Airplane Painters                                  11-27

11-9        Analysis cf Paint Used by Painters                           11-29

11-10       Hematologic Examination of 8C9 Rotogravure Workers           11-33

11-11       Renal Function Investigations of Glue Sniffers               11-42

11-12       Toluene Induced Metabolic Acidosis                           11-44

11-13       Frequency of Lens Changes and Distribution by Exposure
              Time in 69 Age-Matched  Pairs of Car Painters and
              Railway Engineers                                          11-50

12-1        Acute Effects of Toluene                                     12-2

12-2        Subchronic Effects of Toluene                                12-7

12-3        24 Month Chronic Exposures of Fischer 344 Rats Exposed
              6 Hours/Day. 5  Days/Week, to Toluene by Inhalation         12-22

12-4        24 Month Chronic Exposure of Fischer 344  Rats Exposed
              6 Hours/Day, 5 Days/Week, to Toluene by Inhalation         12-23

12-5        Behavioral Effects* of Toluene                                12-36

12-6        Central Nervous System Effects of Toluene                    12-40

12-7        Myelotoxicity Effects of  Toluene                             12-44

12-8        Weekly Blood Picture of Normal Rats  and Rats Exposed to
              600 and 2500 pptn of Toluene 7 Hours/Day, 5 Days/Week,
              for 5 Weeks                                                12-47
                                      xiii

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                            LIST UK TABLES  (cent.)

Number

13-1        Uptake of Toluene in Thin and Obese Her. During Exposure
              to a Toluene Concentration of 375 rag/m^ (100 ppm)          13-6

13-2-s        Partition Coefficients for Toluene at 37'C                   13-12

13-3        Toluene Concentration in Workplace Air and Peripheral
              Venous Blood of Exposed Workers                            13-34

14-1        Epidermal Tumor Yield in 20 Week Two-Stage Experiments       14-4

14-2        Miorobial Mutagenicity Assays                                14-6

14-3        Rat Brine Marrow Ceil Aberrations Following Intraperitoneal
              Injection of Toluene                                       14-11

14-4        Frequency of Unstable and Stable Chrososoa« Changes and
              Chromosome Counts in Subjects Exposed to Benzene or
      '        Toluene or Both                                            U-13

14-5        Effect of Occupational Toluene Exposure and Smoking on
              Chromosomal Aberrations and Sister Chrotcatid
              Exchanges                                                  14-14

14-6        Chromosome Aberrations in Rotoprinting Factory Workers       14-16

14-7        Teratcgenicity Evaluation of Toluene in CFY Rats and
              CFL.P Mice                                        ,          14-20

14-8        Teratogenic Effects of Exposure to Toluene, Benzene,  and
              a Combination of Toluenp and Bsrzene in CFY Rats           14-23

14-9        Teratogenioity and Reproductive Performance Evaluation
              in Rats Exposed to Toluene                                 14-25

16-1        Concentrations of Toluene in Stoppered Flasks                16-3

16-2        Toxic Effects of Toluene to Algae                            16-7

16-3        Toxic Effects of Toluene Vapor on Carrots, Tomatoes,
              and Barley                                                 16-9

17-1        Acute Toxicity of Toiuens to Fish and Aquatic
              Invertebrates                                              17-5
                                      xiv

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                                LIST OF FIGURES

Number                                                                        Page
 6-1        Proposed Reaction Pathways of Toluene Under
              Atmospheric Conditions                                         6-4

 6-2        Microbial Metabolism of Toluene       •                          6-15

12-1        Toluene Levels in Tissue and Behavioral  Performance           12-33

13-1        Metabolism of Toluene in Humans and Animals                   13-18

16-1        Phytoplankton Growth in Vrrious Concentrations of Toluene      16-4

16-2        Growth of Chi orella vulgaris in Medium Containing Toluene      16-6
                                       xv

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                     AUTHORS,  CCOTRIBUTORS, AND REVIEWERS


     This health assessment  document  for  toluene  was prepared -by the Syracuse

Research Corporation under  contract  with U.S.  EPA  Environmental Criteria and

Assessment Office (Mark Greenberg, Project Manager).

     The following scientific staff members of the Syracuse Research  Corporation

(Syracuse,  New  York)  are  listed, by  chapter,  as  principal  and contributing

authors .


     Chapter 1:        Executive Summary

     Chapter 2:        Introduction

     Chapter 3:        Physical and Chemjcal  Properties
                       Author:  Dipak K. Basu

     Chapter M:        Production, Use, and Releases  to  the Environment
                       Author:  Dipak K. Basu

     Chapte" 5:        Abatement Practices in Industry
                       Author:  Dlpak K. Basu

     Chapter 6:        Environmental  Fate, Transport  and Persistence
                       Prinicipal Author:   Dipak K.  Basu
                       rer.trit*;tor:  Arthur Roser.bcrg

     Chapter 7:        Environmental  and Occupational Concentrations
                       Author:  Dipak K.. Basu

     Chapter 8:        Analytical Methodology
                       Author:  Dipak K. Basu

     Chapter 9:        Exposea Populations
                       Author:  Dipak K. Basu

     Chapter 10:       Estimate of Human Exposure
                       Author:  Dipak K. Basu

     Chapter 11:       Effects on Humans
                       Author:  Stephen J. Bosch

     Chapter 12:       Animal Toxicology
                       .Principal Author:  Ethel Ryan
                       Contributor:  Stephen J.  Bosch

     Chapter 13:       Phannacokinetic Considerations in Hucans and  In Animals
                       Author:  Joan T. Colman
                                      xv i

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     Chapter 1U:        Carcinogenic!ty,  Mutagenicity, and Teratogenicity
                       Author:   Stephen  J.  Bosch

     Chapter 15:        Synergisms and Antagoni3iD3  at the Physiological Level
                       Author:   Ethel Ryan

     Chapter 16:        Ecosystem Considerations
                       Author:   Knowlton Foote

     Chapter 17:        Effects  on Aquatic Species
                       Author:   Richard  H.  Sugatt

     Chapter 18:        Health Effects Summary
                       Prinicipal Author:  Patrick Durkin
                       Contributor:   Stephen  J.  Bosch
     The  following individuals  reviewed  portions  of the  document  during its

preparation and  provided  valuable  comments.   Individuals  are listed in alpha-

betical order.

Dr.  Nancy  Adams,  Health Standards  Division,  Occupational  Safety  and Health
     Administration, Washington ,  DC.

Dr.  Michael  Bolger,  Division of   Toxicology,  Food and  Drug Administration,
     Washington, DC.

Amy   Borenstein,   Office  of   Program   Management   and  Evaluation,  U.S. EPA,
     Washington, DC.

Josephine Brecher, Criteria and Standards Division,  Office  of Water  Regulations
     and Standards, U.S. EPA,  Washington, DC.

George Cushraac, Department of  Transportation,  Washington,  DC.

Arnold Edelman, Office of Toxics  Integration,  U.S.  EPA,  Washington,  DC.

Dr.  Penelope  Fenner-Crisp, Criteria and  Standards Division,  Office  of  Drinking
     Water, U.S. EPA, Washington, DC.

Dr.  John  Fowle, Reproductive   Effects  Assessment  Group,  Office  of  Health and
     Envirotnental Assessment,  U.S.  EPA,  Washinton,  DC.

David Friedman, Office of Solid Waste,  U.S.  EPA, Washington,  DC.

Frank  Gostoraski,  Office  of  Water  Regulations   and  Standards,  U.S.  EPA,
     Washington, DC.

Alan  Jennings,  Office  of  Program  Management  and   Evaluation,  U.S.  EPA,
     Washington, DC.
                                     xvii

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Frank Kirvan,  Office of Air Quality Planning and Standards, U.S.  EPA, Durham, NC.

Stephen Kroner, Monitoring  and  Data Support  Division, Office of Water Regula-
     tions and Standards, U.S. EPA,  Washington,  DC.

Dr. Donna Kuroda,  Reproductive  Effects  Assessment Group, Office of Health and
     Environmental Assessment, U.S.  EPA,  Washington,  DC.

Wanda LeSleu-Biswas,  Office of Solid Wast", U.S.  EPA, Washington, DC.

Craig McCormack, Office of Toxic Substances,  U.S.  EPA,  Washington, DC.

Dr. Thomas McLaughlin, Exposure Assessment Group,  Office  of Health and Environ-
     mental Assessment, U.S. EPA,  Washington, DC.

Audrey McBath, Industrial Enviro;nnental Research Laboratory,  Cincinnati, OH.

Dr. Lakshrr,: Misl'jra, Consumer Products Safety  Commission,  Bethesda, MD.

Dr. Debdas  Mukerjee,  Enviror/aentai  Criteria  and Assessment Office,  U.S.   EPA,
     Cincinnati, OH

Dr. Dharm Singh., Carcinogen Assessment Group, Office of Health and Environmental
     Assessment, U.S. EPA, Washington, DC.

Michael Slinak, Monitoring and  Data Support  Division, Office of Water Regula-
     tions  and Standard.**, U.S. EPA, Washington,  DC.

Dr.  Douglas L.  Smith, National Institute  for  Occupational Safety and Health,
     Cincinnati, OK.

Wade Talbot,  Office of Health Research,  U.S.  EPA,  Washington,  DC.
                                     xviii

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                            SCIENCE ADVISORY BOARD
                        ENVIRONMENTAL HEALTH COMITTEE

     The substance of this  document  was  independently peer-reviewed in  public
session by the Environmental Health  Committee,  Environmental  Protection
Science Advisory  Board.

Chairman ;  Environmental  Health
     Dr. Roger 0.  McCleilan,  Director of Inhalation Toxicity Research" Institute,
     Lovelace Bioraedical and Environmental Research Institute,  Albuquerque,  New
     Mexico  87H5

Acting Director, Scier.ee Adv. sory Board
     Dr- Terry F. Yosie, Science Advisory Board, U.S.  EPA,  Wasnington,  DC  20*160

Metspers
     Dr-  Herman  E.  Collier,   Jr.,   President,  Moravian  College,  Bethlehem,
     Pennsylvania 18018

     Dr. Morton Corn,  Professor  and  Director,  Division  of  Environmental Health
     Engineering, School of Hygiene  and  Public  Health, The Johns Hopkins Univer-
     sity, 615 N. Wolfe Street, Baltimore, Maryland  21205

     Dr. John  Doull,  Professor of  Pharmacology  arid Toxicology, University  of
     Kansas Medical Center, Kansas City. Kansas  66207

     Dr. Edward F.  Ferrand,  Assistant Commissioner for  Science and Technology,
     New York City  Department  of  Environmental Protection,  51  Astor Place,  New
     York,  New York  10003

     Dr. Herschel E. Griffin,  Associate Director and Professor of Epidemiology,
     Graduate School of Public Health,  San Diego  State University,  San Diego,
     California  92182
                                     xix

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     Dr.  Jack D. Hackney, Chief, Environmental Health Laboratories,  Professor of
     Medicine,  Rancho  Los Amigos Hospital Campus of the University of Southern
     California, 7601  Imperial  Highway, Downey,  California  902^2
   \
     Dr.  D.  Warner North, Principal,  Decision Focus, Inc.,  5 Palo Alto Square,
     Palo Alto,  CaJ ifornis
     Dr.  William  J.   Schull,  Director  and  Professor of  Population Genetics,
     Center for Denographic and Population Genetics,  School  of Public Health,
     University of Texas  Health Science  Center  at Houston, Houston, Texas 77030

     Dr.  Michael  J.  Syssons, Professor,  Department  of Bi ostatistics,  School  of
     Public Health,  University  of  North Carolina,  Chapel  Hill, North Carolina
     2771 ',

     Dr.   Sidney  Weinhouse,  Professor  of Biochemistry,  Senior  Member,  Pels
     Research  Institute,   Temple University  School  of  Medicine,   Philadelphia,
     Pennsylvania 191 <*0
Consultant
     Dr.  Bernard  Weiss,  Division  of  Toxicology,  University of  Rochester  of
     Medicine,  Rochester,  New York

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                             1.   KXF.CUTIVE SUMMARY

1.1.   ENVIRONMENTAL SOURCES,  FATE.  AND LEVELS  '
     Toluene, a  homolo?  of t^en'/ene  that  contains a single methyl groop,  is  a
clear, colorless liquid at rcxim temperature.  The molecular formula of toluene is
C_H  and -the molecular weight  i.4 Q2.13.  The structural formula is given below.
     Other physical properties  of  toluene include a melting point  of  -95°C,  a
boiling point of 110.6°C, a flash poin", of i4.44°C,  a  vapor pressure of 28.7 torr
at 25°C, and a density of 0.8669 g/mZ  at  20°C.   Toluene is  slightly soluble ir.
both fresh and salt water (535 mg/2 and 379 mg/S., respectively) at a temperature
of  25°C.    The  physical  properties  of  toluene  indicate that  toluene in  the
environment  is  likely to  be present  in  the  air, and  that  toluene originally
present  in  water may  be  transferred to  the  atmosphere.   Toluene  can  undergo
photochemical  reactions,  psrticularly  under  atmospheric smog conditions.   In
aqueous media under the conditioas of water chlorination, toluene may be chlor-
inated  followed  by subsequent  hydrolysis to  benzaldehyde.   This  reaction may
account for the benzaldehyde  detected  in  some finished drinking waters.
     The general population may be exposed to  toluene through inhalation of air,
ingestion of  food  or water,  or dermal  exposure.   The four  largest sources of
emission of  toluene  to the atmosphere  are, in  descending order of importance,
automobile use, industrial use of  toluene  as a solvent, coke ovens, and toluene-
producing industries.  In addition to air, toluene has  been detected in drinking
water  and  in the  flesh  of  edible  fish.   Dermal  exposure  to  toluene  occurs
pritaarily in the workplace.  The estimated quantities of toluene  taken in by the
general public from each  source are between a  trace and 9** mg/week by inhalation
(depending on whether an individual  resides in an urban or rural  area or near an
industry  that  uses  toluene)  and  0.0  to 0.75 mg/week  from  food and  water.
Occupational exposure  (up to  18,000 mg/week) or  cigarette  smoking (14 mg/week
from  140 cigarettes)  will   increase   an  individual's  exposure  to  toluene.
Although there  are technical  problems with  estimating  inhalation exposure to
toluene, there is reasonable agreement  between the  values obtained  by dispersion
                                       1-1

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modelingand those obtained from calculations using monitoring data.  It should be
noted that the exposure scenario discussed above  does not account for inhalation
and  dermal  exposure   to  toluene   from   gasoline   during  vehicular  filling
operations, or frotu. solvents or other toluene-containing consumer products.  No
quantitative estimate  of  either  the  number of people exposed or the  extent  of
exposure  can   be  provided  for  these  sources,   although  consumer  usage  could
contribute significantly  to total exposure.
     The  total amount  of «-.oluene produced  in   the  United  States  in  19?8 was
3595 million k<3-  The majority (96.5?)  is  produced by catalytic reformation from
selected  petroleum fractions,  and  the  remainder  is  produced  from  pyrolytic
cracking,  and  as a recovered  by-product   of  styrene  production and  coke  oven
emission.  This value of 3595  million kg is for isolated toluene  and accounts for
only 115 of the total toluene produced;  the remaining 89? of the toluene produced
is  not  isolated as  pure toluene  but  is a  benzene-toluene-xylene mixture used in
gasoline.  Toluene  is  also  used  as feed stock for the production of benzene and
other chemicals,  as a  gasoline additive, and as  a solvent.
     Activities  associated  with automobiles  (marketing and evaporation of gaso-
line and automobile exhaust) are the  largest single atmospheric source of toluene
release  (677  million kg/year),  with  industries  using toluene as a  solvent  (the
paint  and  coating,  adhesive,  ink,  and  pharmaceutical  industries)  being the
second  largest source of toluene in the atmosphere (375 million  kg/year).  These
two sources account for 75? of the toluene emitted to the atmosphere.  The amount
of  toluene released to  other media in  the environment  is  small  and is equal to
approximately  0.15? of  the  total  amount released to the atmosphere.
     The preferred method for the monitoring of  toluene in ambient air consists
of  sorbent  collection,  tnermal  elution,  and GC-FID determination.   For a 25  I
sample, the detection  limit is  <0. 1  ppb  (0.38 ug/nH).   Purge and ~,rap with GC-
photoionization  detection is the most widely used method  for  the analysis of
toluene in aqueous samples.  With a 5 m£ sample,  the  method  has a detection  limit
of  0.1 ppb (0. 1 ug/2,).
     Toluene is the most  prevalent aromatic hydrocarbon in the  atmosphere, with
average  measured levels  ranging from O.'M  to  59  ppb (=0.53  to  200 ng/rn^).
Toluene  has  also  been detected  in  surface  waters and in  treated wastewater
effluents at levels generally below 10 ppb  (10 \ig/H). A concentration of toluene
as  high as 19 ppb (72 ug/2.) has been detected in a  drinking  water supply.  In  a
study of toluene  levels in  the  tissue  of  edible aquatic  organisms,  95$ of the
                                       1-2

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samples contained less than 1 ppm (1 mg/kg) of toluene.   The atmosphere is the
major  environmental  receiver  for  toluene.     It  has  been  estimated  that
approximately 124 million people in the U.S. are exposed to atmospheric toluene
at a concentration level greater than 0.27 ppb (1.0 ng/nr) •
     Toluene released  to the aquatic  or soil environment  is at  least  partly
removed by biodegradation.  There  is  little information on the rate  and extent of
biodegradation in  soil:  however,  in one study,  a  half-life of between  20 and
60 min was  observed in  soil  containing toluene-degrading  bacteria,  and  in  a
second study  20  to  60?  of toluene  was  removed following  percolation  through
140 era of sand.  Because of the limited number of studies available, the extent
of toluene degradation in soil cannot be determined, although studies  with pure
cultures indicate that a variety of bacteria and  fungi can use toluene, and some
pure cultures have been isolated that can use toluene as  a sole source of carbon.
Toluene is also readily  biodegraded  in aqueous media, both in surface  water and
during  wastewater treatment;  however,  disappearance of  toluene  from  aqueous
media  is mainly  through  evaporation  and  transport to  the atmosphere.   The
conversion of  toluene to compounds  that can  be  used as sources of carbon and
energy suggests that toluene will be degraded rapidly by microbial species pro-
liferating at the expense of the  compound, and will  not accumulate significantly
in the environment.
1.2.  EFFECTS  ON HUMANS
     Toxicity  studies  of humans have  primarily  involved  evaluation of indivi-
duals exposed to toluene via inhalation in experimental  or occupational settings
or during episodes of  intentional abuse.  The results of these studies indicate
that although  people exposed occupationally may be  at risk, exposure to ambient
levels of toluene is not likely to constitute a significant hazard to the general
population.
     The health effect of primary concern is dysfunction of  the central nervous
system  (CMS).  Acute experimental  and  occupational  exposures to toluene in the
range  of 200 to  1500  ppm  (=750  to  5600 mg/nr)  have elicited dose-related CNS
alterations  such  as  fatigue,  confusion,  and  incoordination,  as   well  as
impairments  in reaction time  and  perceptual  speed.    Following  initial CHS
excitatory   effects   (e.g.,   exhilaration,    lightheadedness),   progressive
development   of   narcosis   has  characterized  acute  exposures  to  excessive
concentrations  of   toluene   (i.e.,  levels   approaching   the  air  saturation
concentration  of   approximately  30,000 ppm   ( = 113,000   mg/m )).    Repeated
                                       1-3

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occupational exposures to  toluene' over a period of  years  at levels of  200 to
400 ppm  (=750  to  1500 pg/m3) have  resulted  in  some  evidence  of  neurologic
effects,  and  chronic  exposure  to  mixtures  of  solvent  vapors  containing
predominantly  toluene at  levels  of  30  to  100 ppm  (=100  to 100 mg/m3)  have
resulted  in impaired  performance  on  tests  for  intellectual and  psychomctor
ability and muscular  function.   Prolonged abuse of  toluene or solvent  mixtures
containing toluene have,  on occasion, led to residual or permanent CNS  effects.
     Early  reports  of occupational  exposures  ascribed  myelotoxic effects  to
toluene, but the majority of recent evidence indicates that toluene is  not toxic
toward the blood or bone  marrow.  The myelotoxic effects previously attributed to
toluene currently are considered to have  been the result of concurrent  exposure
to  benzene,  which  was typically present  as a  contaminant.   Acute exposures to
toluene have not resulted  in  any definite effects on  heart  rate  or blood pres-
sure.
     Liver enlargement was reported in an  early study of  painters exposed to 100-
1100 ppm (=400 to 4100 mg/m ) toluene for  2 weeks to more than 5 years,  but this
effect was not associated with clinical evidence of liver disease or corroborated
in  subsequent  studies.   Chronic occupational exposure to  toluene  or  intensive
exposure  via glue or  thinner sniffing generally  has not  been  associated with
abnormal  liver function.  Evidence of  renal  dysfunction  has been observed  in
workers who were accidentally overexposed to toluene  and in toluene abusers, but
studies of workers exposed to 100  to  1100 ppm  (=400  to  4100 mg/m  ) toluene for
2 weeks to 5 years and 60 to 100  ppm  (=200 to 400 mg/m3)  toluene for over 3 years
did not report  abnormal urinalysis findings.   Several  reports  have  appeared
recently  that   associate  deliberate  inhalation  of  toluene  with metabolic
acidosis.
     Subjective  complaints  of dysmenorrhea have been reported by  women exposed
for over 3 years t.o 60 to 100 ppm (=200 to 400 mg/m ) toluene and concommitantly
to  20 to 50 ppm gasoline  in a "few" working places.  Disturbances of menstruation
have  also  been  reported in  female  workers  exposed  concurrently to  toluene,
benzene, xylene, and  other unspecified solvents.  The limited available data do
not,  however,  specifically  associate  occupational  exposure  to  toluene  with
menstrual effects.  Information on the possible reproductive effects of toluene
in  males is not available.
     Single  short-term exposures  to moderate levels  of toluene  have,  on occa-
sion, been reported to cause transitory eye and respiratory tract irritation, but
                                      1-4

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irritative effects have generally not  been observed  in workers  exposed repeti-
tively to toluene*  Dermal contact with toluene may cause skin damage due to its
degreasing action.
1.3.  ANIMAL STUDIES
     The most  pronounced  effect of  toluene  in animals is  on  the CNS.   Acute
exposure via  inhalation to  high  levels  of  toluene  has  been  associated  with
depression of activity.   Levels below  1000 ppm (=3800  mg/m^) have little or no
effect on gross manifestations of  behavior,  although more  sensitive  methods of
assay (i.e., detection of changes  in cognition •arid brain neuromodulator levels)
have indicated effects at lower levels.
     Early  studies  with animals suggested thdt  toluene induced  myelotoxicity,
but most studies that used toluene that contained.negligible amounts  of benzene
have not produced injury on blood-forming organs.
     Inhalation of  concentrations  of up to  1085  ppm (=^100 mg/m  ) toluene  for
6 weeks  or  300 ppm  (=1100 mg/m^)  for  24 months,   or  ingestion  of  590 mg
toluene/kg  body weight/day  for 6  months produced no evidence of  liver damage;
however, several studies noted an increased liver weight or slight histological
changes suggestive  of possible  liver damage  at higher  levels  of  vapor  exposure
(=2000 ppm  (=7500 mg/m  ) in rats), or in animals  treated by the  intraperitoneal
route (=0.i) g/kg).
     Renal  injury was  noted  in rats,  dogs,  and guinea pigs after  subchronic
inhalation  of  toluene  vapors at  levels  in  excess of  600 ppm (=2300 mg/nr) in
three studies, but  other subchronic  exposures  in which rats, dogs, guinea pigs,
and  monkeys inhaled (concentrations up to 1085 ppm  (=U100 mg/mjj) or  ingested
(590 mg toluene/kg  body weight/day)  toluene did not produce **enal damage.
     Although no effect was observed on the lungs  of rats, guinea pigs,  dogs, or
monkeys after intermittent exposure  to  1085 ppm (=4100 mg/m )  toluene vapor for
6 weeks,  in rats  after inhalation  of  up to 300  ppm ( = 1100 mg/m  ) toluene  for
2U months,  or  in  rats  after  ingestion of 590 mg toluene/kg body weight/day for
6 months, other studies  have  noted irritation effects  in the respiratory tract in
dogs, guinea pigs, and rats.  Ser.sitization of the heart in mice, rats, and dogs
has  been associated with inhalation  of toluene.
     The acute oral toxicity (LDr )  of toluene in rats  is in the range of 6.0 to
7.5 g/kg, which indicates only slight toxicity in  this  species.  Inhalation LC<-Q
values have been  reported  in the  range of 500 to 700 ppm (=1900 to 2600 mg/nr)
for mice and 4000  ppm ( = 15,000 mg/m  ) for rats.  Acute dermal toxicity appears to
                                       1-5

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be tjuite low (rabbit LD,-0 of  12.2  g/kg),  but  slight  to moderate irritation has
been noted in rabbit and guinea pig skin and in rabbit  corneas after application
to the skin and eye, respectively.
1.14.  ABSORPTION, DISTRIBUTION, METABOLISM, ELIMINATION, AND RELATED
        PHARMACUK1NETICS
     Toluene is readily absorbed from the respiratory tract. Studies with humans
indicate that the total amount of toluene absorbed is  proportional to the concen-
tration of toluene in inspired air, the length of exposure, and pulmonary venti-
lation, which in turn depends upon the level of physical activity.  Approximately
50? of the amount inspired is retained in the body.  Absorption of toluene from
the gastrointestinal tract is probably fairly complete, based on excretion data
from  experimental  animals.   Toluene is  absorbed  less  readily through the skin
than through the respiratory or gastrointestinal tracts.
     Animals  given  toluene  orally or by  inhalation  had  high  concentration0  of
toluene in their adipose tissue and bone marrow, and moderately high concentra-
tions  of  toluene and  its metabolites  in liver and  kidney.   These  results are
reasonable  based on tissue-blood  partition  coefficients  and known  routes  of
metabolism and  excretion.
      The initial step in the metabolism of toluene is  side-chain hydroxylation by
the  hepatic mixed-function oxidase system,  followed  by  oxidation to  benzoic
acid.   Benzoic acid is then  conjugated  with  glycine to form  hippurlc acid and
excreted in  the urine.   In  both humans and animals,  60 to 75$  of  the absorbed
toluene  can  be accounted for as  hippuric acid  in the  urine,  regardless of the
dose  or whether the chemical was administered orally or by inhalation.  Much of
tne  remaining toluene  is exhaled  unchanged.   The  excretion  of toluene and its
metabolites  is  rapid; the major portion occurs within  12 hours of oral adminis-
tration or the  end of  inhalation exposure.
1.5.   CARCINOGENICITY, MUTAGEN1CITY, AND  TERATOGENICITY
      Inhalation  exposure  to   toluene  at  concentrations  of  up  to  300 ppm
(=1100 mg/m  )   for  24 months   did  not   produce  an  increased  incidence  of
neoplastic,  proliferative,  inflammatory,  or  degenerative  lesions in  various
organs of rats relative to unexposed controls.   It should be noted, however, that
the 300 ppm (=1100 mg/m )  exposure did  not represent a maximum tolerated  dose.  A
bioassay  of  commercial  toluene in  rats and  mice  exposed  via inhalation  is
currently  being conducted  by  the  National  Toxicology  Program Carcinogenesis
Testing Program.  Other studies  indicate  that toluene  is  not  carcinogenic when
applied topically to the shaved skin of laboratory animals and that it does not
                                      1-6

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promote the development of skin tumors following initiation with 7,12-dimethyl-
benz[a]anthracene.
     Toluene  has  been  shown to  be  non-mutagenic  in  a battery  of  microbial,
nanmalian cell, and whole organism test systems,  but assays for sister-chromatid
exchange  (SCE)  and  cytogenetic   effects  nave  provided conflicting  results.
Increased frequencies of SCEs and/or chromosome aberrations were found in some,
but not all, studies of lymphocytes from  workers  who were chronically exposed to
toluene, and SCEs-and/or aberrations were  not induced  in Chinese hamster ovary
cell.3 or human lymphocytes exposed to toluene in culture.  Russian studies have
reported  chromosome  aberrations   in  the  bone  marrow  cells  of rats  exposed
subcutaneously and  via  inhalation to  toluene, but  these findings have not been
corroborated  in  other studies  of rats  following intraperitorieal injection  of
toluene.
     Toluene has been reported to  induce cleft palates and embryotoxic effects in
mice following oral exposure, but  it was  not  teratogenic  in mice or rats follow-
ing  inhalation exposure.   Embryotoxic effects (increased incidence  of skeletal
anomalies  and signs  of  retarded  skeletal  development,  low  fetal weights)  and
increased  maternal  toxicity were  noted, however, in  some  of the rats and mice
exposed  via inhalation.
 1.6.   EFFECTS  ON  ECOSYSTEMS
     The ecological  effects  of  toluene have  been investigated using aquatic and
terrestrial  microorganisms,  aquatic  invertebrates, fish,  and  higher  plants.
Toluen-e  can both stimulate and inhibit growth of bacteria and algae, depending on
the  species  and the  concentration of toluene.   The growth  of  most  species of
bacteria,  algae,  and other  microorganisms is not  inhibited  until  the toluene
concentration  exceeds  10  to TOO mg/H,    Toluene is  acutely toxic  to  aquatic
invertebrates  and fish  at  concentrations ranging between 3 and  1180 mg/i.  The
lowest  concentration shown  to  cause  sublethal  effects  in  aquatic  animals was
2.5 mg/JL.  Chronic  toxicity data  are  available for two  species  of fish;  marine
sheepshead minnows were affected at toluene concentrations of 7.7 mg/8,  but  not at
3.2 mg/i, and fathead minnows were affected at 6  rag/it but not at ^ mg/8,.  Chronic
effects occurred  at  concentrations that were  about  2 to  18  times lower than the
acute LCj-Q for these si.'cies, indicating that chronic effects may occur at lower
levels in more sensitive species.  Toluene concentrations between 0,1 and  1.0 ppm
have been reported occasionally in surface  waters (0.1 to 1.0 mg/X.) and sediments
(0.1 to  1.0mg/kg).   These  concentrations are sufficiently close to the toxic

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concentrations for  sensitive  species to  indicate  that acute or  chronic  toxic
effects may occur in some polluted habitats, especially after accidental spills
of toluene.  Toluene has  only a low bioconcentration potential and is metabolized
and rapidly  depurated  from  fish, which  indicates  that toluene is  unlikely  to
biomagnify through aquatic food webs. Toluene, however, has been shown to impart
an unpleasant  taste to  fish  that  inhabit contaminated water.   The impact  of
toluene spills or chronic low-level  pollution on ecosystems  is unknown.  Adverse
effects may  occur  but  probably are  limited  by rapid  rates of loss  of  toluene
through evaporation and  biodegradation.
1.7.  HEALTH EFFECTS SUMMARY
     Considerable information  is available  on the  effects  of toluene on humans
and experimental animals after inhalation exposures.  The data on oral  exposure
are much less satisfactory, although one acceptable subchronic oral study using
rats is available.  No  information on dermal  exposures suitable for use in numan
risk assessment was encountered.
     Based on a few studies involving controlled exposures of humans to toluene
vapors  as  well  as some  reports  of  occupational  incidents  and  voluntary  abuse
("glue  sniffing"),  the  dose-response  relationships  for  the acute  effects  in
huaans  of  single short-tenn exposures to toluene can be estimated as:

      10,000  to 30,000  ppm         :    Onset  of  narcosis  within  a  few
      (=38,000 to  113,000 mg/m  )        minutes.  Longer exposures may
                                       be lethal.
      >'i,000  ppm                   :    Would  probably cause rapid
      (=15,000 mg/m  )                   impairment of reaction time and
                                       coordination.  Exposures of one
                                       hour  or longer might lead to
                                       narcosis and possibly death.
      1,500 ppm                    :    Probably not lethal  for exposure
      (=5600  mg/nr)                     periods of up to eight hours.
     300 to  800 ppm               :    Gross  signs of incoordination
      (=1100  to 3000 mg/m )             may be expected during
                                       exposure periods up to
                                       eight  hours.
     400 ppm                      :    Lacrimation and irritation
     (=1500  mg/m )                     to the eyes and throat.
     100 to  300 ppm               :    Detectable signs of  incoordination
     (=400 to 1100 mg/m  )              may   be  expected  during  exposure
                                       periods up to eight hours.
                                       1-8

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     200 ppm                      :     Mild throat  and eye  irritation.
     (=750 mg/m3)
     50 to 100 ppm                :     Subjective complaints
     (=200 to 400 mg/m3)                (fatigue  or  headache)
                                       but  probably no
                                       observable impairment
                                       of reaction  time or
                                       coordination.
     >37 ppm                      :     Probably  perceptible to most
     (=150 mg/m-0                      humans.

Because of the deficiencies in the studies on which these estimates are  based, as
well as variations in sensitivity to  toluene that  may be expected in  the human
population, these estimates should be regarded as approximations  only.
     The  subchronic  and  chronic  inhalation  data  lend  themselves less to the
definition  of  dose-response  relationships.   Most  of the  reports  on human
exposures failed  to  define precisely  levels or  durations of  exposure,  involved
relatively small numbers  of exposed individuals, and did not  adequately control
exposure to other toxic agents.  The animal data are of little use in supporting
the human  data  because-humans  appear  to  be mpre sensitive  to toluene  than the
experimental  animals on which data are available.
     Qualitatively, dermal exposure to toluene can  cause skin damage,  as is the
case with many solvents,  but systemic signs of  intoxication are likely to occur
only in cases of gross overexposure.

1.8. RESEARCH NEEDS
     Although  the available  data  from  human  and  animal  toxicologic  studies
indicate that  ambient  exposure to toluene does not currently present  a human
health hazard,  it is apparent  that further investigation  is  needed in several
areas.    Some of the most  important areas In which incomplete  information  is
available, and  that  should  be considered in foitnulating  research  needs, are
represented below.  This list  of research  needs, however,  is  not structured  in
terms  of  relative priorities  among the various areas of investigation noted
below.
    1.   Monitoring data.  Up-to-date monitoring data pertaining to atmos-
         pheric levels around point sources involving solvent use,  ambient
         air levels in rural  and remote areas, and drinking water levels are
         needed to more accurately evaluate human exposure to toluene.
                                     1-9

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2.   Consumer exposure.   General population  exposure  to toluene from
     gasoline usage/spillage during vehicular filling operations, from
     use of  paint  and varnish thi nners/removers, arid  from  the use of
     other toluene-containing  consumer products  rtamina unevaluated,
     although exposure  from  these  sources could  substantially cont-
     ribute to total exposure.  Data are needed on the magnitude, fre-
     quency,  duration, and extent of exposureCs) from these sources to
     properly assess general population toluene exposure.

3.   Neurobehavioral  toxicity.    It  is evident  that  the CNS  is most
     sensitive to the effects of toluene.  Although  the effects of acute
   '  high level  toluene exposure are fairly well do direr) ted, there is a
     paucity of data regarding the behavioral aned to be detenr' ,ed to
     properly evaluate potential  risk fr-e ambient exposure.

!*.   Carcinogenic! ty.   Although  it  is  improbable  that  toluene  is  a
     strong carcinogen, the possibility that it  is a weak one cannot be
     excluded  on  the  basis  of  the  information  that   Is  presently
     available.   The results of  the ongoing NTD  carcinogenesis bio-
     assays  cf  toluene  should  help resolve  the  issue,  and  mitigate
     concern for the apparent  deficiencies  cf the CUT  (198C) bioassay.

5.   Mutagenicity.   Toluene has  been  shown to be  non-rautagenic in  a
     variety  of  microbial and raamialiar;  systems,  but the  results  of
     cytogenetic   assays   (sister-chromatid  exchange  and  chromosome
     aberration)  are conflicting.   Additional  testing  is needed  to
     resolve  the  possibility  that toluene may  be  a weak clastogen or
     mutagen.

6.   Teratoeenicity.   Toluene has been reported  to  be  teratogenic to
     mice  fallowing  oral  exposure 'Nawrot  and Staples,  1979),  but not
     to mice or rats  following inhalation  exposure (Huddle and Ungvary,
     1978; Litton  BJonetics,  Inc.,  1978b; Tatrai et  al.,  1980).  The
     uncertainty  over  the teratogenicity  of toluerte should serve as  a
     stimulus  for  further  research,   including  evaluation  of  the
     association  between   teratogenic  effects  ap.d  chronic lew level
     exposure to toluene.

7,   Reproductive  effects.   Th^  reports  of  dysraenorrhea in   female
     vrorkers   (Matsushita  et  al.,   1975),  degeneration  of  germinal
     epithelium in the testes of  male  rats (Matsushita et al.,  1971),
     and increased  follicle-stimulating  hormone (FSH) levels  in rats
     (Andersson et  al.,   1980)  that have  been  associated with toluene
     exposure suggest that  the  reproductive effects  of  this  compound
     should also  be  considered  in formulating research  needs.   .Again,
     this should include evaluation of effects associated with chronic
     low level exposures  to toluene.

8.   Respiratory Defense  Mechanism.   Various  gaseous  air pollutants,
     (e.g., nitrogen oxides and ozone)  have boen demonstrated to  affect
     respiratory  tract  defense  mechanisms,  resulting  in  increased
                                 1-10

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susceptibility to respiratory infectic/i in some mammalian species
(e.g., rat, mome) studied.   Complex  dose-response  relationships
have been demonstrated, wnereby specific patterns of duration and
frequency  of exposure,  as  well  as   the  concentration  of  the
particular gaseous air pollutant, appear to be important factors.
To date, little or no information has  been published in regard to
the evaluation of potential.effects  of  volatile organic substances
such  as  toluene  on  respiratory   defense  mechanisms.    It  is
recommended that such studies be conducted in the near future.
                            1-11

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                               2.  INTRODUCTION

     EPA's Office of Research and Development has prepared this health assess-
ment to serve  as a "source document" for Agency use.  The scope of this docu-
ment addresses  toluene  in relation to  the  total  environment.   It is expected
that this document will serve the information needs of many government agencies
and private  groups that may be  involved  in decisionmaking activities related
to toluene.
     In the development of the assessment document, existing scientific litera-
ture has been  surveyed  in detail.  Key  studies have been evaluated and summary
and conclusions  have been prepared so that  the chemical's  toxicitv and related
characteristics  are qualitatively identified.
     The present  document represents an up-to-date evaluation of the available
'toluene data base.  The document assesses  all major sources of toluene in the
environment,   general   ambient   concentrations  representing  potential  human
exposure  levels,  and health effects demonstrated  to  be  associated with expo-
sure  of  man or  lower organisms.  More  detailed, updated evaluations regarding
sources,  emissions,  ambient air  concentrations,  and public  exposure levels
will  be  carried out in  the future should EPA decide  to undertake  specific
regulatory  action(s)  for  toluene.
      The  information  found  in  this document is  integrated  into  a  format designed
as  the  basis  for  performing  risk assessments.  Where  appropriate,  the  authors
of  the  document  have  attempted to  identify  gaps in  current,  knowledge  that
limit risk  evaluation capabilities.
                                      2-1

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                     3.  PHYSICAL AND CHEMICAL PROPERTIES

     Toluene is a homolog of  benzene in which one hydrogen atom has been replaced
by a methyl  group.   Some of the relevant  physical  and chemical  properties  of
toluene are described below.

3.1.   SYNONYMS AND TRADE NAMES
     Methacide
     Methylbenzene
     Methyl benzol
     Phenylraethane
     Toluol

3.2. IDENTIFICATION NUMBERS
     Chemical Abstracts Service (CAS) No.:  108-88-3
     Registry of Toxic Effects of Chemical Substances (RTECS)  No.:  XS5250000

3.3.  STRUCTURE, MOLECULAR FORMULA,  AND MOLECULAR WEIGHT
                                   CH.
     Molecular Formula:  ^,. Hg
     Molecular Weight:  92.13

3.4.  PHYSICAL PROPERTIES

3.4.1.  Description.  Toluene is a clear, colorless liquid at ambient temperature
that has  a benzene-like odor.   It is  both  volatile and  flammable  (Windholz,
1976) .

3.4.2.  Other Physical Properties.
     Melting Point  (Weast, 1977):                     -°5°C
     Boiling Point  (Weast, 1977):                     110.6°C
     Density (g/m2., 20°C) (Weast,  1977):             0.8669
     Specific Gravity (15.6/15.6°C) (Cier, 1969):    0.8623
     Vapor Pressure (25°C) (Weast, 1977):            28.7 torr
     Vapor Density  (air   1)  (Weast, 1977):          3.20
                                                           •
                                      3-1

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     Vapor Pressure (25°C) (Weast,  1977):
     Vapor Density (air   1)  (Weast,  1977):
     Percent in Saturated Air
     (760 mm, 26°C) (Walker,  1976):
     Density of Saturated Air-Vapor
     Mixture (760 mm (air   1).
     26°C) (Walker, 1976):
     Solubility (Sutton and Calder, 1975):
        Fresh water (25°C)
        Sea water  (25°C)
     Flammable Limits (percent
     by volume in air) (Walker, 1976):
     Flash Point (closed cup)  (Walker, 1976):
     Autoignition Temperature  (Walker, 1976):
     Log Octanol-Water Partition
     Coefficient (Tute,  1971):
     Odor Threshold in Air (Walker, 1976):
        Coke derived
        Petroleum  derived
     Surface Tension  (20°C) (Walker,  1976):
     Liquid  Viscosity (20°C)  (Walker, 1976):
     Refractive Index (68°F)  (Cier, 1969):
     Conversion Factor (in air, 25°C):
28.7 torr
3.20
1.09

534.8 mg/i
379.3 Bg/i

1.17 to 7.10
40°F
552°C

2.69

4.68 ppm
2. 14 ppm
28.53 dynes/cm
0.6 cp
1.49693
1 ppm =_ 3.77 mg/m3
1 mg/nr = 0.265 ppm
3.4.3    Significance  of  Physical  Properties  with  Respect  to  Environmental
Behavior.  The .volatility of toluene, as indicated by its relatively high vapor
pressure,  indicates  that a  substantial fraction  of environmental  toluene  is
likely  to  be present  in the vapor  phase mixed with air.   The  relatively high
volatility of toluene,  combined  with  its low solubility in water,  may  lead  to
intermedia transfer of toluene from water to the air phase.  The details of the
environmental fate of toluene as  determined  by  its  physical and chemical proper-
ties are discussed in  Chapter 6.
     The log octanol-water partition  coefficient  for toluene  may have signifi-
cance in determining its affinity toward organics in  soil and aquatic organisms.
The details of the  bioconcentration factor for toluene based on the octanol-water
partition coefficient  value are discussed in Chapter 9.  The knowledge of physi
                                      3-2

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cal properties such  as flammable limits and flash  point is important for the safe
handling  and  transport  of  toluene;  data on  density  and  solubility may  be
necessary for health effect studies.
3.5.  CHEMICAL PROPERTIES
     Toluene  undergoes  substitution  reactions,  either  on  the  aliphatic  aide
group (-CH-,) or on  the  benzene  ring.   These substitutions occur exclusively at
the ortho (2) and para
                           positions marked in the following figure:
     Nitration,  sulfonation,  halogenation, methylation,  and chloromethylation
 are  some examples of substitution reactions.   These reactions  occur at a rate
 between  2.1  and  467  times  faster with toluene than with benzene (Cier, 1969).
     The methyl  group  in toluene  is susceptible  to dealkylation  to  produce
 benzene  (Bradsher, 1982).
                                       Thermal
                                          or
                                      Catalytic
                                                                     CH,
      At one time,  the most significant use of toluene was in the production of
 benzene by  the  above  reaction  (Cier,  1969).
      Toluene undergoes a reversible disproportionation and transalkylation reac-
 tion  in the presence  of a  catalyst  (Cier,  1969).
                                                             CH.
     Hydrogenation  of toluene takes place readily to produce methylcyclphexane
 (Cier,  1969).
               n
                              +   H_
                                        catalyst
o
                                       3-3

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     The reverse process of dehydrogenation of methylcyclohexane is the princi-
pal mode of toluene manufacture.  Methylcyclohexane is found in  petroleum frac-
tions, along with other naphthenes (Cier, 1969).
     Oxidation of  toluene  under catalytic  conditions yields benzoic acid as  a
principal product  (Cier, 1969).
              n
                                        catalyst
     Chlorinati-on of  toluene  under  actinic  light  conditions  yields  methyl  sub-
stitution products (Cier, -1969) .
                   hv
                                                      HC1,
     The hydrolysis of benzalchloride produces benzaldehyde (Gait, 1967)
     The above reactions may have some significance with respect to chlorination
 of .drinking water.  The oxidation of toluene that occurs in drinking water may be
 one  of the sources  of  benzaldehyde  and benzoic  acid  detected in drinking water
 (U.S. EPA, 1980).
     In  the  presence  of  catalysts and  in the absence of  light,  chlorination
 produces o- and £-chlorotoluene  (Cier, 1969).
     In the vapor phase, toluene is relatively unreactive toward RO radicals and
0, found in the troposphere.   It is, however, relatively more reactive toward OH
radicals;  the  products of the  reaction are normally  benzaldehyde  and cresols
(Brown et al.,  1975).   This  reaction  may  have  significance with respect to the
fate of toluene in the  atmosphere and is discussed in detail in Section 6.1.

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     Toluene forms azeotropes with a number of solvents, including paraffinics,
naphthenics, and alcoholic hydrocarbons,  Azeptropes are important in the puri^
fication of toluene, in solvent technology, and in the recovery of toluene from
reaction mixtures (Cier, 1969).
     Toluene is  marketed as  nitration  grade  (1°, boiling  range  of 1°C), pure
commercial grade (2°C), and all other grades.  Generally accepted quality stan-
dards for the first two grades are given by the  American  Society for Testing and
Materials (Cier,  1969).  The actual concentration of toluene  is  not stipulated in
these  specifications;  however,  the nitration  grade  (1°)  and pure commercial
grade (2°)  toluene  are of 99.5 to 100?  and 98.5  to  99. 'J? purity, respectively
(USITC,  1979).   All other grades include toluene and are used as solvent grade
and  for  blending aviation and  motor  gasoline.    The  non-fuel  toluene  (solvent
grade) is of 90  to  98.4$ purity  (USITC,  1979).
     Commercial  toluene may  contain  benzene as  an impurity.   Therefore,  all
health  effect  studies involving  toluene  should  specify  the quality of toluene
used for experimentation.   If benzene  is  present  in the  toluene,  it  must be
demonstrated  that  the observed health  effects  are  not wholly or partly due to
benzene.  Because  of  this contamination, it may also be necespary to determine
the  amount  of benzene released  to the  environment  due  to industrial usage of
toluene.
      In  general, toluene is  quite stable in air,  and most of the chemical reac-
tions  discussed  above require specialized conditions.   While some of the reac-
tions  may have environmental  significance, the  majority of the chemical reac-
tions  discussed  above  are  conducted under conditions of  commercial and  research
applications.
 3.6.  REFERENCES

 BRADSHER,  C.K.   (1982).   Toluene.   In:   McGraw-Hill  Encyclopedia  of  Science  and
 Technology,  5th ed.  Vol.  13.,  McGraw-Hill Book  Co.,  NY.   p.  759-760.
                                       3-5

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BROWN, S.L., CHAN,  F.Y., JONES, J.L., LIU, D.H., MCCALEB,  K.E.,  MILL,  T.,  KAPIOS,
K.N.,  and SCHENDEL, D.E. (1975).  Research Program on Hazard Priority Ranking  of
Manufactured  Chemicals,  Phase II—Final  Report,  chemicals  1i-20.   Prepared  by
Stanford  Research   Institute,  Menlo  Park,  CA.    National  Science  Foundation,
Washington,  D.C.    Available from:    National Technical  Information Service,
Springfield,  VA (NTIS PB 263  161).

CIER,   H.E.   (1969).   Toluene.    In:    Kirk-Othmer Encyclopedia  of Chemical
Technology, 2nd ed.  Standen, A..  Editor. New York:   John Wiley  and  Sons, Inc.,
Vol. 20, p. 528.

GAIT,   A.J.  (1967).  Heavy Organic Chemicals.   Oxford, England:   Pergamon Press,
Ltd.,   249  pp.

SUTTON, C. and J.A. CALDER, 1975.   Solubility  of Alkylbenzenes in Distilled Water
and Seawater  at 25°C.   J.  Chem. Eng.  Data, 20:  320-322.

TUTE,  M.S.   1971.   Principles  and  Practice  of Hansch Analysis:  a guide  to
Structure-activity Correlations  for  the  Medical  Chemist.    Adv.  Drug Res.,
5:  1-77.

U.S.  EPA.   1980.   Ambient  Water  Quality Criteria for Toluene,  Office of Water
Regulations  and   Standards,  Criteria   and   Standards   Division.     U.S.   EPA,
Washington, DC.  Available ,from NTIS,  Order No.   PB  81-117855,  Springfield, VA.

USITC (UNITED  STATES   INTERNATIONAL  TRADE  COMMISSION).    (1979).    Synthetic
Organic Chemicals:   United  States Production  and  Sales,  1973,  USITC  Publication
1001,  USITC,  Washington, DC.  20^36.

WALKER, P.-  1976.  Air Pollution Assessment  of Toluene.   Report prepared by Mitre
Corporation.   Prepared  for U.S.   Environmental  Protection  Agency.    Available
through NTIS  Order  No.  PB 256735,  Springfie.IJ, VA.
                                       3-6

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WEAST,  R.C.,  Ed.    (1977).    CRC  Handbook of  Chemistry and  Physics,  58th ed.
Cleveland, OH:  Chem.ical Rubber Co.

WINDHOLZ, M.,  Ed.   (1976).   The Merck Index:  An Encyclopedia of Chemicals and
Drugs,  9th ed.  Rahway, NJ:   Merck and Co.,  Inc.
                                       3-7

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              4.   PRODUCTION,  USE,  AND RELEASES  TO THE ENVIRONMENT

14.1.  MANUFACTURING PROCESS TECHNOLOGY
     Toluene is produced  primarily  from three sources:  (1) petroleum refining
processes,  (2)   indirectly   as  a  by-product   of   styrene   production,   and
(3) indirectly as a by-product  of coke-oven operations.
4.1.1.  Petroleum Refining Processes.  Low levels of toluene  are present in crude
p^roleu-.  Toluene is produced from petroleum by two  processes:  (1) catalytic
reforming and (2) pyrolytic cracking.
     4.1.1.1,  CATALYTIC REFORMING —  The largest quantity of toluene  produced in
the United  States is  generated in the  catalytic  reforming  process.   The total
estimated toluene produced  in  this process  in  1978  was 3110 million kg.  This
represents about 87% of the total amount of toluene produced  in the United States
in 1978  (fable 4-1).
     Catalytic  reforming involves  the  catalytic dehydrogenation  of selected
petroleum fractions that  are  rich  in  naphthenic hydrocarbons to yield a mixture
of aromatics  and  paraffins.   The proportions of aromatics  and paraffins in the
reformate  depend upon the  feedstock  used  and the  severity of  the reforming
operation (Cier,  1969).  At  present, reforming operations are geared  primarily to
producing a benzene-toluene-xylene (BTX)  reformate   from which  the  individual
aromatics are recovered  (.Cier,  1969).  Toluene is isolated from  the reformate by
distillation, followed by washing with sulfuric acid  and redistillation.  Only a
small  fraction  of  catalytic  refcrmate,  however,  is utilized  for isolating
toluene.  The unseparated toluene in  catalytic  refornate is used for gasoline
blending.
      4.1.1.2.  PfROLYTIC CRACKING — The second largest quantity of toluene comes
from  pyrolytic cracking.  Of  the total isolated toluene produced in  the United
States in 1978, approximately 9J  (324  million kg)  was obtained from  this source
(Table 4-1).
     When heavier hydrocarbons, such  as  hydrocarbon condensates, naphtha, and
gas  oil, are pyrolytically cracked  for the  manufacture of olefins, pyrolysis
gasoline is produced as a by-product.  The amount of  pyrolysis gasoline produced
from  pyrolytic   cracking depends  on the   feedstock  and the  manufacturing
conditions  (Mara  et al.,  1979). The  by-product,  pyrolysis  gasoline,  contains  a
high percentage of aromatics.   Toluene  can be isolated from pyrolysis  gasoline by

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                                   TABLE 4-1
                 U.S. Production of  Isolated Toluene  in  1978a
Production
Process
Catalytic reforming
Pyrolytic cracking
Styrene by-product
Coke oven by-product
TOTAL
Amount
Produced
(10b kg)
3110
324
135
26b
3595
Percent
of Total
86.5
9
3.8
0.7
100
aSource:  Little, 1981
 This value does not include toluene obtained from tar distillers.
                                      4-2

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distillation, removal of any olefins and diolefins,  and  redistillation.  Not all
pyrolysis gasoline produced  in  the  United States is used for the production of
isolated toluene.
4.1.2.   By-Product  of  Styrene  Production.   When  styrene  is  produced  by the
dehydrogenation  of ethylbenzene',  some toluene  is  also  synthesized as  a by-
product.  The toluene isolated from the by-product is  unsuitable for  chemical or
solvent use.  Therefore,  toluene obtained from  this  source  is  used either for
gasoline  blending or as  feed for  the  manufacture  of  benzene  by  the hydrode-
alkylation process (Mara et  al.,  1979).  In 1978,  approximately  135 million  kg of
isolated toluene, which was about 4J of the total, was obtained  as the by-product
of styrene production (Table  4-1).
4.1.3.  By-Product of Coke-Oven  Operation.  The  production of coke by the  high-
temperature  carbonization of coal  yields  coal-tar  and  crude light  oil  as by-
products; both of these  by-products  contain toluene.  The production of toluene
from  'distillation of  coal-tar  is  minimal  (Mara et al., 1979);  however,  some
toluene is isolated  from  crude light  oil.  As shown in  Table 4-1, approximately
26  million  kg of  toluene were  isolated from coal-derived toluene  in the year
1978.   This  amounted to about 0.7? of the total, isolated toluene produced during
the same  year.
4.2.  PRODUCERS
      Of the  total  toluene produced in the  United States for internal consumption,
only  about   11$  is isolated  as  toluene  (Table  4-2).   The  remainder stays in
gasoline  as  a benzene-toluene-xvlenp  (BTX) "ixtur".   TV.c Luuai  amount,  of toluene
available in the  United States in 1978, both  isolated  and  non-isolated, is  shown
in  Table  4-2.
      The  identification of isolated  toluene producers,  their estimated toluene
producing capacity,  and the estimated  amount of toluene  produced  in 1978 from
catalytic  reforming, pyrolytic  cracking, and styrt.r"?  by-product  are shown in
Tables  4-3,  4-4,  and  4-5.    The identification  of the  producers  of isolated
toluene  from coke-oven   by-product  is  given  in Table 4-6;  the  capacity for
isolated  toluene  production  and the  actual amount  of toluene  produced are not
given  because the data  are  unavailable.   It  should be  pointed out that many
producers captively  consume  the  toluene that they produce.
      During  1979, the  production  of toluene  from  coke-oven   operators  had  a
reported  increase  of 17.6$ over  1978 (USITC,  1979).  The production  of toluene
from  petroleum refiners has  been  reported to have decreased  by  4.3J  during the
                                       4-3

-------
                                   TABLE 4-2

                  Isolated and Non-Isolated Toluene Available
                        in the United States in 1978a
Source
Catalytic reforming
Pyrolytic cracking
Styrene by-product
Coke oven by-product
Imports
Exports
SUBTOTAL
TOTAL

Isolated
3,110
324
135
26
192
-364
3,^23

Quantity
(10fc kg)
Non-Isolated as BTX
27,000
197
NA
96
NR
27,293
30,716
 Source:  Little, 1981
NA = not applicable, NR = not reported
                                     *-«»

-------
                           TABLE 4-3




Producers of Isolated Toluene from Catalytic Reforming in 1978a
Company and Location
Amerada Hess - St. Croix, VI
American Petrofina - Big Spring. TX
Beaumont , TX
Ashland Oil - Catlettsburg, KY
N. Tonawanda, NY
Arco - Houston, TX
Wilmington, CA
Charter Oil - Houston, TX
Coastal States - Corpus Christi, TX
Commonwealth - Penuelaa, PR
Crown - Pasadena, TX
Exxon - Baytown, TX
Getty - Delaware City, DE
El Dorado, KS
Gulf - Alliance, LA
Philadelphia, PA
Port Arthur, TX
Kerr McGee - Corpus Christi, TX
Marathon - Texas City, TX
Mobil - Beaumont , TX
Monsanto - Chocolate Bayou, TX

Pennzoil - Shreveport, LA
Phillips - Sweeney, TX
Guayama, PR
Quintana-Howell - Corpus Christi, TX
Shell - Deer Park, TX
Toluene
Capacity
(10b kg)
460
164
125
99
39
125
49
39
56
395
46
411
b
20
194
92
49
148
72
280
33
c

33
335
56
197
Isolated Toluene Produced
(105 kg)
310
110
84
67
26
8U
33
26
3b
» 266
31
277
NA
13
130
62
33
100
49
189
22

NA
22
226
38
133
                              1-5

-------
                              Table 4-3.   (cont.)
V
Company and Location
Sunoco - Corpus Christ!, TX
Marcus Hook, PA
Toledo, OH
Tulsan, OK
Termeco - Chalmt-tta, LA
Texaco - Port Arthur, TX
Wesr-ville, NJ
Union Oil - Lemont , IL
Union Pacific ~ Corpus Christi, TX
TOTAL
Toluene
Capacity
(10b kg)
138
151
2U7
66
115
92
T32
56
99
M613
Isolated Toluene Produced
(106 kg)
93
102
166
*Ul
78
62
89
38
67
3108
 Source:  Little, 1981
 1980 capacity for this producer was 85 million kg.
 I960 capacity for this producer was 72 million kg.
NA = not applicable.

-------
                                   TABLE

             Producers  of Isolated Toluene frora Pyrolyaia Gasoline'"1



                                       Toluene
                                       Capacity       Isolated Toluene Produced
Company and Location                   (105 kg)                (10° kg)


Arco - Chanelview, TX                    105                       76

Commonwealth - Penuelas,  PR               ^9                       36

Dow - Freeport, TX                        13                        9.1)

Gulf - Cedar Bayou, TX                    66                       IS

Mobil - Beaumont, TX                      16                       15

Monsanto - Chocolate Bayou, TX           132                       96

Union Carbide - Taft, LA                  66                       48

     TOTAL                               UH7                      328,4
aSource:  Little,  1981
                                     K-7

-------
                                  TABLE 14-5




            Producers  of Isolated Toluene froa .Styrene By-Product3
Company and Location
American Hoechst - Baton Rouge, LA
Arco - Beaver Valley, Ih
Cos-Mar - Carville, LA
Dow - Freeport , TX
Midland, MI
El P-A3Q Natural Gas - Odessa, TX
Gulf - Donalasville, LA
Monsanto - Texas City. TX
Standard Oil (Indiana1) -
'I'exas "ity, TX
SJHOCO - Corpus Christi , TX
U.S. Steel - Houston, TX
TOTAL
Styrene
Capacity
(105 kg')
uoo
100
590
660
UO
68
270
630
380

36
54
3)400
Isolated Toluene Produced
(10" kg)
16
U
24
26
5.
2.
11
27
15

1.
2.
134.




5
1




14
2
8
Source:  Little,  1981
                                    U-8

-------
                                   TABLE U-6

      \ Producers of  Isolated  Toluene from Coke-Oven Crude Light Oils3
          Plant
          Annco                                 Middletown, OH

          Ashland Oil                           Catlettsburg, KY
                                                N. Tonawanda, NY

          Bethlehem Steel                       Bethlehem, PA
                                                Sparrows  Pt., MD

          CF and I                              Pueblo, CO

          Interlake                             Toledo, OH

          Jones and Laughlin                    Aliquippa, PA

          Lone Star                             Lone Star, PA

          Republic Steel                        Youngster, OK
                                                Cleveland, CH

          U.S. Steel                            Clairton, PA
                                                Geneva, UT



aSource:  Little, 198l

-------
same period  (USITC,  1979).   This  resulted  in  a net  decrease  of  't.2S  in the
overall  isolated  toluene  production  in 1979 as compared to  1978  (Table li-1)
(USITC, 1979).
U.3. USERS
     As mentioned in Section li.2.,  most of  the toluene produced as BTX mixture is
never  isolated  but  remains  in  various refinery  streams  for use  in  gasoline.
Isolated  toluene,  on  the  other  hand,  is  used  for  different  purposes;  the
consumption of isolated  toluene  in  different  usage  is  shown  in Table  ^-7.  The
fluctuating, but  largest, single use of isolated toluene  is in the production of
benzene through the hydrodpalky]ation (HDA) process.  The  fluctuation in the use
of  isolated toluene exists because the HDA process is used as an  effective means
of  balancing supply  and  demand  for  benzene  (Mara et al.,   1979).    The U.S.
producers  of  benzene through  the  KDA process,   their  capacity,  and the amount
produced are shown  in Table  .-8.
     The second  largest  use of isolated toluene is back-blending into gasoline
for increasing  the  octane ratings.   Approximately  1^65  million  kg of isolated
toluene,  representing  35.1?  of  1978 consumption, was used  for  gasoline back-
bler.ding.
     The third  largest use of toluene is in solvent applications, with the major
use being  in the paint and coatings industry.  Significant amounts also are used
in  adhesives,  inks, Pharmaceuticals, and  other  formulated  products.   With the
establishment  of  federal and  state  laws limiting the  emission  of  aromatic
solvents  in the workplace  and in the general environment,  the demand for toluene
as  a solvent  has declined  significantly  (amount unspecified)  since 1975  (Mara
et  al.,  1979).    Identification  of  specific  users  of toluene as  a solvent is
difficult  because the users  are  toe  widespread.
     Another major use of isolated toluene is as  a raw material in the production
of  toluene diisocyanate  (TDI), benzyl  chloride,  benzoic acid, xylene, and vinyl
toluene.  Manufacture of phenol,  cresols,  toluene sulfonic acids, nitrotoluenes,
terephthalic  acid,  caprolactam,  and  styrene  are some  of  the  minor  uses  of
isolated  toluene  (Mara  et al.,   1979).    A  small  amount of  isolated toluene
(6.6 million kg,  <1J of  total) is  used for the manufacture of p_-cresol (Little,
1981).   The  latter  compound is  used primarily for  the  manufacture  of the-
pesticide 2,6-di-tert-butyl-p-cresol (BHT).  Judging from the percent of toluene
used in  the  manufacture  of  BHT,  its  emission  from this manufacturing process
should be considered insignificant.
                                      lJ-10

-------
                                    TABLE I*-7




      Consumption of Isolated and Non-Isolated Toluene in Different Usages'
Usage
Non-isolated:
Gasoline as BTX
Isolated:
Benzene dealkylation
Gasoline back-blending
Solvent for paint and coatings
Solvent for adhesives, inks,
and Pharmaceuticals
Toluene diisocyanate
Xylene
Benzoic acid
Benzyl chloride
Vinyl toluene
Miscellaneous others
Net export
TOTAL
Amount Used /year
(10° kg)

27,293

1,675
1,465
263

132
200
98
65
36
25.
39
172
4,170
Percent of
Total Use in
Each Category

100

40.2
35.1
6.3

3.2
4.8
2.4
1.6
0.9
0.6
0.9
4.1
100.1
aSource:   Little, 1981

-------
                                   TABLE H-8




      Consumers  of Toluene  for  the  Manufacture of Benzene by HDA Process3
Company and Location
American Petrofina - Port Arthur, TX
Big Spring, TX
Ashland Oil - Catlettsburg, KY
Coastal States - Corpus Christi, TX
Commonwealth - Penuelas, PR
Crown - Pasadena, TX
Dow - Freeport, TX
Gulf - Alliance, LA
Philadelphia, PA
Monsanto - Alvin, TX
Phillips - Guayama, PR
Quintana-Howell - Corpus Christi, TX
Shell - Odessa, TX
Sunoco - Corpus Christi, TX
Toledo, OH
Tulsa, OK
TOTAL
Toluene Used Benzene Production Capacity
(1(T kg) (10b kg)
59
103
9V
156
298
59
65
122
52
103
103
191
18
52
163
39
1674
77
130
120
200
380
77
84
160
67
130
130
250
23
67
210
50
2155
aSource:  Anderson et al., 1980
                                         1-12

-------
     The identification 9f primary users of toluene as a chemical intermediate,
their production capacity, and  the amount produced is shown in Tables 4-9 and
4-10.  It should be pointed out that the amount of isolated tolueqe used in the
United States in 1978  (excluding  net export) was 4000 million  kg according to
Table 4-7.  However, Table 4-2 shows that the total amount of toluene available
for internal consumption during the  same period (excluding net export) was only
3600 million kg.  This discrepancy is due to the fact  that  Table 4-7 is based on
data that are only estimates, and  the  data  in Table  4-2  were obtained from the
manufacturers  who  reported  their net  toluene production  to the  U.S.  Inter-
national Trade  Commission.
4.4. ENVIRONMEOTAL  RELEASE
     The three  primary sources of toluene release or emission to  the environment
are  production,  usage,  and inadvertent sources.   In  addition to these anthro-
pogenic  sources, some  toluene  is released  into the  environment  from natural
sources.
4.4.1.   Emission  from Production Sources.   Toluene  can  be  released into the
environment  during its production as  process losses, fugitive  emissions, and'
storage losses.  Process emissions are those  that originate from the reaction and
distillation  vents  deliberately  usad  for  venting gases.    Storage  emissions
.originate  from  losses during  loading and  handling of  the product  used for
manufacturing processes and storage of the final product.  Fugitive emissions are
those  that  have their  origin  in plant equipment leaks.  The air  emission factors
used to estimate the total omission of toluene from different  production sources
have been obtained  from  Mara et al. (1979) and  the  values  are given in  Table
4-11.
      Based  on the emission factors indicated in Table  4-11,  the amount  of toluene
emitted into  the atmosphere  from  the four production  sources  has been estimated
in Table  4-12.    Atmospheric releases  of  toluene from  each  source  shown  in
Table  4-12  are  from production of both isolated and non-isolated toluene.   It  is
assumed that  the air emission is dependent only"on the manufacturing process and
is the  same for both isolated and  non-isolated  toluene from  the  same  process.
     The manufacturing processes may lead also to toluene release in other media.
The  release  of  toluene in water from petroleum refineries performing catalytic
reforming and pyrolytic  cracking processes  is assumed to  be  negligible because
the  concentration of toluene  has  been  determined  to be below the quantification
limit in more than  90? of  discharged water  from the refineries  (Little,  1981).
                                      4-13

-------
                                   TABLE 4-9




               Producers of Toluene Diisocyanate  (TDI) in  19?8a
Company and Location
Allied Chemical - Moundsville, WV
BASF Wyandotte - Geismar, LA
Dow Chemical - Freeport, TX
Du Pont - Deepwater, KJ
Mobay Chemical - Baytown, TX
New Martinsville, WV
Olin - Astabula, OH
Lake Charles, LA
Rubicon Chemical - Geismar, I A
Union Carbide - S. Charleston, WV
TOTAL
TDI Capacity
(10° kg)
36
45
15
32
59
45
14
45
18
25
364
Toluene Used
(106 kg)
20
25
25
17
32
25
7
25
10
13
199
aSource;  Mara et al.,  1979
                                     4-14

-------

Other Toluene

Company and Location
Arco - Houston, TX
Sunoco - Marcus Hook, PA
TOTAL
Kalaraa - Kalama, WA
Monsanto - St. Louis, MO
Velsical - Beaumont, TX
Chattanooga, TN
Pfizer - Terre Haute, IN
Tenneco - Gar field, NJ
TOTAL
Monsanto - Bridgeport, NJ
Sauget , IL
Stauffer - Edison, NJ
UOP - E. Rutherford, NJ
TOTAL
Dow - Midland, MI
TABLE 1-10
Chemical Intermediate Users in
Production
Capacity
(10° kg)
Xvlene Producer's
89
92
181
Benzoic Acid Producers
64
5
23
27
3
7
129
Benzyl Chloride Producers
36
36
5
1
78
Vinyl Toluene Producers
27

1978a

Toluene Used
(10b kg)
48
50
98
33
2
12
14
1
3
65
16
16
3
0.5
35.5
25
Source:  Mara et al., 1979
                                      1-15

-------
                                   TABLE 4-1
             Toluene Air Emission Factors from Production Sources
                                         Emission Factor
                                      (kg lost/kg produced)
Source
Process
Storage
Fugitive
Total
Catalytic reforming    0.00002

Pyrolytic cracking     0.00015

Styrene by-product     0.00001

Coke oven by-product   0.00050
               0.00006

               0.00060

               0.00060

               0.00060
               0.00002

               0.00015

               0.00015

               0.00015
               0.0001

               0.0009

               0.00076

               0.00125
 Source:  Kara et al.,  1979
                                          1-16

-------
                                   TABLE  14^12

                  Estimated  Atmospheric Toluene  Emissions  from
                        Four Major Production Sources
Production Source
  Total Amount
    Produced
(million kg/yr)
 Total        Total
Emission     Emission
 Factor    (103 kg/yr)
Catalytic reforming  - Isolated         3,110
                     - Non-isolated    27,000

Pyrolytic cracking   - Isolated           324
                     - Non-isolated       197

Styrene by-product                        135

Coke oven by-product - Isolated            26
                     - Non-isolated        96

     TOTAL
                     0.0001


                     0.0009


                     0.00076


                     0.00125
              3,011


                469

                103

                153

               3,736

-------
     Coking operations, however, can lead to toluene release in other media.  The
toluene-containing wastewaters  from  coking  plants  that  originate  from waste
ammonia  liquor,   final cooler  blow  down,  and  benzol plant  wastes  have  the
following distribution (Little,  1981):
          Direct discharge:  33$
          Publ?cly Owned Treatment Works  (POTW):  25%
          Quenching:   40$
          Deep well injection:  2%
     Two-thirds of  the wastewater  from the quenching operation is recirculated
and  actually not  discharged.    Therefore, only  73$  of  the  total  wastewater
containing toluene is  actually  discharged  to the environment.
     The average  volume of effluents  produced from coke-oven operation (Little,
1981),  the  toluene concentration  in  these  effluents (Little,  198D,  and the
emission factors  in these effluents are  given in Table 4-13.
     For a total coke  production of 44 x  10^ kg  in 1978 (Little, 1981),  the total
                                                                       Q
amount  of toluene discharged in wastewater is  calculated  to be 44 x 10  x 4.43
x  10~   x 0.73 = 142 x   10  kg.  Some toluene in wastewater may finally  enter other
media,  because  wastewater from the  quenching  operation is sent  to  sumps that
generate  only  solid  and  gaseous  wastes  (Little,   1981).    Therefore,  the
distribution  of total  released  toluene in  untreated wastewater can be  estimated
as  given in  Table 4-14.
4.14.2.  Emission  from  Toluene Usage.  The emission of toluene from  various usages
has been estimated  from  emission  factors and  the amounts  used.  The values for
the emission  factors  obtained from Mara  et al.  (1979)  are shown in Table 4-15.
     The  atmospheric   emission  of  toluene  from  its  production sources, such as
gasoline  in  non-isolated  BTX  and  the isolated form  (for  back-blending),  has
already been  included  in Table fc-12.  The emission  factor  for miscellaneous uses
has been assumed  to be the average of other toluene usages excluding its use as  a
solvent.   All the  toluene  used in paint  and  coatings has been  assumed to be
ultimately  released  to the  atmosphere   (Mara  et al.,  1979).    Therefore,  an
emission factor of 1.0 has been  estimated  for this usage.   Fifteen  percent  of the
toluene used  as a solvent for adhesives,  inks,  and Pharmaceuticals is  recovered
for fuel use  (Mara  et al.,  1979);  the remainder  is  emitted to the  atmosphere.
Hence,  an emission  factor of 0.85  has been assumed for this usage.
     Based on the emission  factors given  in Table  4-15,  the estimated  toluene
emissions from its various usages  are shown in  Table  4-16.
                                      4-18

-------
                                   TABLE U-13

                                                                       a
       Toluene  Emission Factors in Wastewater from Coke Oven Operation
Effluent
Liters of Effluent
 Produced/kg Coke
Toluene
 Con/i
(mg/J.)
  Emission
   Factor
(kg/kg coke)
Waste ammonia liquor

Final cooler blow down

Benzol plant wastes

    TOTAL
       0.16

       0.13

       0.20
   3-1

  17.0

   8.6
  0.496 x 10

  2.21  x 10
                                                                          -6
  1.72 x 10
                                        x  10
-6

-6
 Source:  Little,  1981

-------
                                  TABLE 11-14

       Toluene Released  in Different Media  from  Coke-Oven  Wastewater3
Medium
Air
W^ter
Land
POTW
Percent of
Total Released
20
33
22
25
Amount released/yr
(103 kg)
28
^7
31
36
Toluene releases fros quenching are arbitrarily assumed to be evenly distr
buted between land and air.

-------
                                  TABLE H-15
                    Toluene Emission Factors  for  Its  Uses'
Emission Factor
(kg lost/kg used) ,
Usage
Benzene production
Solvent for paint
and coatings
Solvent for adhesives,
ink, Pharmaceuticals,
and others
Toluene diisocyanate
Xylene production
Eenzoic acid
Benzyl chloride
Vinyl toluene
Miscellaneous
Process
0.00005
NA
NA
0.00077
0.00005
0.00100
0.00055
0.00055
NA
Storage
0.00010
NA
NA
0.00032
0.00010
O.OOOHO
0.00030
0.00030
NA
Fugitive
0.00005
NA
NA
0.00019
0.00005
0.00010
0.00015
0.00015
NA
Total
0.00020
1.0
0.85
0.00128
0.00020
0.00150
0.00100
0.00100
0.00100
 Source:  Mara et al., 1979
NA = not applicable.
                                        H-21

-------
                  TABLE H-16




Estimated Toluene Emission from Different Uses
Source
Benzene production
Solvent for paint and
coatings
Solvent for adhesives,
Pharmaceuticals, and
others
Toluene diisocyanate
Xylene production
Benzoic acid
Benzyl chloride
Vinyl toluene
Miscellaneous others
TOTAL
Amount Used/yr
(10b kg)
1675
263
inks ,
132
200
98
65
36
25
39
2533
Emission
Factor
(kg lost/kg used)
0.0002
1.0
0.85
0.00128
0.0002
0.00150
0 . 00 1 0
0.0010
0.0010

Total Emission/yr
(103 kg)
335
263,000
112,000
256
20
98
36
25
39
375,809
                  H-22

-------
     It can  be  concluded from  Table  4-16 that, among  the  different  usages of
toluene, the maximum emission  (excluding  inadvertent  sources)  occurs  from sol-
vent application (see Section 4.14.3.).
     The released toluene from the  different user sources shown in Table 4-16 has
been assumed to  enter only  one  medium,  air.   The use of  toluene as  a solvent,
however,  has been  found  to  produce toluene  'in  wastewater   (Little,  1981).
Table 4-17 shows the total estimated release of toluene to  aqueous media from its
use as a solvent in different industries.
4.4.3.  Emission from Inadvertent Sources.   Because gasoline  consumes  a vast
amount  of  total  toluene  produced  (Table  4-7),  this use  constitutes the largest
source of  environmental  emission of toluene.   The  emission  of toluene from its
use in gasoline  can occur from three distinct  sources:   evaporation from its use
in the automobile, evaporation  from marketing activities  (handling and transfer
of bulk quantities), and emission  from automobile exhaust.
     Other inadvertent sources of toluene emissions into the environment include
transportation spills into surface water and land,  other manufacturing processes
not producing toluene, different combustion sources, and  cigarette smoke (Table
4-18).   The inadvertent release of toluene from other  manufacturing proctjses
occurs primarily from feedstock contamination,  by-product  formation, and the use
of  oil.   An  example of the  latter  source  is in  the manufacture of acrylonitrile
in which wastewater ponds are covered with oil  to control the release of volatile
orgariics.
      The   release  of toluene  into  different  media  from  various  inadvertent
sources is  shown  in Table  4-18.   Intermedia  transfers  of the compound will
possibly change  the emission values given  in Table 4-18 because of the volatility
of  toluene.
4.4.4.     Non-anthropogenic Sources.--  Substantial amounts  of  toluene  can be
released into the environment from petroleum seepage  in the oceans and on land,
and from  the weathering  of exposed coal strata.   However, no estimate of total
environmental release of toluene from these natural sources is available.  Some
vegetations  can  be  natural  sources of  toluene  in  the environment.  Toluene is
naturally  produced by the tropical tolu  tree.   It has  been identified in roasted
filberts,  in peanuts and macadamia nuts^  in grape essence,  and in cooked potatoes
(NRC,  1980).  It is,  however,  unlikely  that  significant  amounts of toluene are
released into the  environment from the  vegetations  in  the  United States (NRC,
1980; Seila, 1979).
                                      4-23

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                                  TABLE 14-17
Toluene Released in Aqueous Media from Use as a Solvent  in  Various  Industries'
Toluene Cone.
in
Wastewater
Source (|ig/Ji)
Ink formulating 1600
Textile products 14
Gum and wood chemicals 2000
Paint formulating 990
Leather tanning 78
Pharmaceuticals 515
TOTAL
Wastewater
Discharged
Percent (10° 8,/d)
Occurrence
87 0.092
i>S 2000
78 0.11
87 2.8
25 200
62 250

Amount of
Toluene
Released
(103 kg/yr)b
0.033
3.8
0.17
0.72
1.2
24
29.9
 Source:  Little, 1981
 Based on 300 operating d/yr-

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                                  TABLE 4-18
             Toluene Emission from Different Inadvertent Sources'
Environmental Release
(103 kg/yr)
Source
Gasoline marketing
Automobile gasoline evaporation
Automobile exhaust
Transportation spills:
Oil
Gasoline
Toluene
Propylene oxide manufacture
Polychloroprene manufacture
Ethylene-propylene rubber manufacture
Ethylene-propylene terpolymer
production
Wood preserving industry
Insulation board manufacture
Hardboard manufacture
Acrylonitrile manufacture
Combustion processes:
Coal refuse piles
Stationary fuel combustion
Forest fires
Agricultural burning
Structural fires
Cigarette smoke
Others
TOTAL
Air
19,000
18,000
640,000

MR
NR
NR
36
460
90

it, 200
NR
NR
NR
59

4,400
13,000
7,000
1,000
<1,000
53
8
703,306
Water
NR
NR
NR

400
680
2.2
NR
NR
NR

NR
6.3
neg.
neg.
NR

NR
NR
NR
NR
NR
NR
NR
1,089
Land
NR
NR
NR

5.6
230
11
NR
NR
NR

NR
NR
NR
NR
NR

NR
NR
NR
NR
NR
NR
NR
247
 Source:  Little, 1981
NR = not reported
                                        4-25

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4.4.5.   Sum of  Emissions from  All Sources.   The  emissions of  toluene into
Different media  from  all sources are given  in Table 4-19.   The estimates also
include toluene emission from coke production.  The emission  of toluene from coke
oven operation is based on an emission factor of 0.00024  (Mara et al., 1979) and
an estimated coke production of 44 x 109 leg  (Little, 1981)  for the year 1978.
     It is evident from Table 4-19 that the toluene released  into the environment
predominantly enters one  medium, the  atmosphere.   The three largest sources of
toluene  emission  in  descending order  are  auto  exhaust,  solvent  use,  and
evaporative loss from automobile  and service stations.  A  large amount of toluene
from  land  and  water   spills  is  also  likely  to  enter air as  a result  of
evaporation.   The large  figure  for the combined  release  of toluene  into the
atmosphere  explains  the reason  for  its  presence as the aromatic hydrocarbon of
highest concentration  in  the ambient atmosphere  (Chapter  7).
4.5. USE  OF TOLUENE IN  CONSUMER  PRODUCTS
     The  consumer  products  shown in Table 4-20  and analyzed prior to 1969 may
contain some toluene.  The percent of toluene  in these products also is indicated
in the same table.  The emission of  toluene into the environment  from this source
is already  included under Section 4.4.2.
      Information available  through  the  Food  and  Drug Administration   (FDA)
(Bolger,  1981) shows that of the  19,500 cosmetic products  registered with the FDA
through August  14,  1979,  664 products contain varying  percents of toluene.  One
of the  products  contains  more than  50?  toluene,  166 products contain 25 to 50?
toluene,  492 products  contain  10  to 25? toluene,  1 product contains  1  to 5?
toluene,  and 4  products  contain 0.1$  or  less toluene.   The  use  of  toluene is
related to  nail  base coats,  nail  enamel, nail polish removers, and other manicure
products.
                                      4-26

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                                  TABLE 4-19




             Total Yearly Release of Toluene into Different Media
Source
Production (see Tables
4-12 and 4-14)
Usage (see Tables 4-16
and 4-17)
Inadvertent (see 'lable
4-18)
'Coke production
TOTAL

Air
3,764
375,809
708,306
10,560
1,098,439
Environmental
(103 kg/yr)
Water
47
30
1,089
NA
1, 166
Release
Land
31
NA
247
NA
278

POTW
36
NA
NA
NA
36
NA = not available.
                                    4-27

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                                   TABLE 4-20
               Consumer Product Formulations Containing Toluene
Product
Percent Toluene Content
    ; China cement, solvent type




     Contact rubber cement




     Microfilm cement, cotton base




     Model cement




     Plastic cement, polystyrene




     Shoe cement




     Tire repair, bonding compounds




     Paint brush  cleane. 3




     Stain, spot, lipstick, rust  removers




     Nail polish




     De-icers, fuel antifreeze




     Fabric dyes




     Indelible inks




     Marking inks




     Stencil inks




     Solvents and thinners
20 to 30




may contain toluene




27 to 30




up to 20 to 25




214




may contain toluene




>80




contain 25 to 90 BTX




may contain toluene




35




30



<_60




may contain' toluene




80 to 90




140 to 60




may contain toluene
 ^Source:   Gleason  et  al.,  1969
                                        4-28

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M.6.  REFERENCES

ANDERSON, G.E,,  LIU,  C.S.,  HOLMAN,  H.Y. and KILLUS, J. P.  (1980).  Human Exposure
to  Atmospheric   Concentrations  of  Selected   Chemicals.   Prepared  by  Systems
Applications, Inc.,  San  Rafael,  CA,  under Contract  No.   EPA 68-02-3066.  U.S.
Environmental Protection Agency, Research Triangle Park, NC.

BOLGER, M.  (1981).  Private communication between M. Green burg", ECAO,  EPA and M.
Bolger, Toxicologist, Food  and Drug  Administration,  Washington, DC.  April 13,
1982.

CIER,  H.E.    (1969).   Toluene.   In:    Kirk-Othmer Encyclopedia  of Chemical
Technology, 2nd ed.  Standen, A.,  Editor.  New  York:  John  wiley and Sons, Inc.,
Vol.   20, p. 528.

GLEASON,  M.N.,  GOSSELIN,  R.E.,  HODGE,  H.C. and  SMITH,  R.P.   1969.   Clinical
Toxicology  of  Conmercial Products:   Acute  Poisoning,  3rd  ed.   Baltimore,  MD:
Williams  and Wilkins, Co.,  pp. VI.1-132.  (Cited in  Slimak,  1980).

LITTLE,  A.D.    (1981).   Exposure  Assessment of  Priority Pollutants:   Toluene.
Draft  report  prepared by Arthur  D.  Little,  Inc.,  Cambridge,  MA,  for the U.S.
Environmental Protection Agency,  Research Triangle Park,  NC.

MARA,  S.J., SO, E,C. and SUTA,  B.E.  (1979).   Uses,  Sources, and Atmospheric
"Emissions of Alkylbenzene  Derivatives,  Final  Report.    Prepared  by SRI  Inter-
national, Menlo Park, CA,  under Contract No. 68-02-2835.   U.S. Environmental
Protection  Agency, Research Triangle Park,  NC.

NRC (NATIONAL  RESEARCH COUNCIL).  (1980).   The Alkyl  Benzenes.   Committee on
Alkyl  Benzene  Derivatives,  Board on Toxicology and  Environment Health Hazards.
Assembly of Life Sciences,   National Research Council.  Washington, DC;  National
Academy  Press.
                                      t-29

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SEILA,  R.L.  (1979).  Non-Urban Hydrocarbon Concentrations in Ambient Air North
of Houston,  Texas.    EPA Report  No.  EPA-600/3-79-010,  Environmental  Sciences
Research  Laboratory,  Office  of Research and  Development, U.S.  EPA,  Research
Triangle Park, NC.  Available through NTIS,  Springfield, VA.  Order No. NTIS PB
293227.

SLIMAK,  M.    (1930).   Exposure  Assessments  of Priority  Pollutants:   Toluene.
Report  (draft)  prepared  by Arthur  D.  Little,  Inc.,  MA.   Prepared  for U.S.
Environmental   Protection   Agency.   Monitoring  and   Data  Support  Division,
Washington, DC.

USITC  (UNITED  STATES INTERNATIONAL  TRADE  COMMISSION).    (1979).   Synthetic
Organic  Chemicals:  United  States Production and Sales, 1978, USITC Publication
1001,  USITC,  Washington, DC 20^36.
                                      1-30

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                      5.  ABATEMENT PRACTICES IN INDUSTRY

     The four major potential sources of toluene release to the environment, in
order of importance (Table U-19),  are  (1) inadvertent sources, such as vehicular
emissions and losses during gasoline transfer,  (2)  solvent use in paint, coating
adhesives, and inks,  (3)  coke  production,  and (4) manufacturing  sites  such as
petroleum refineries and chemical -plants.  Therefore, the institution of pollu-
tion control devices for these four major  sources  can  be expected to produce a
large impact on the overall toluene level  in the environment.
5.1.  ABATEMENT PRACTICES FOR INADVERTENT  SOUBCES
     The two ma lor  sources  of  vehicular  emissions  of toluene in the atmosphere
are  exhaust  emissions  and evaporative  emissions  from  the gas  tank  and  the
carburetor-   Crankcase emissions have been  eliminated  essentially through the
use of positive crankcase ventilation technologies (U.S. EPA, 1980).
   '  The installation  of  catalytic  converters  on automobiles has resulted in a
   i
significant  reduction of hydrocarbon  emissions  from automobiles.   Generally,
tailpipe catalysts control systems remove unsaturated and aromatic hydrocarbons,
including  toluene, more  efficiently than paraffinic  hydrocarbons  (U.S.  EPA,
1980).  Therefore, both the photochemical reactivity and the  mass of hydrocarbons
emitted are  reduced by  the  catalytic  converter systems.
     Evaporative emissions  from automobiles have been reduced through the use of
adsorption  regeneration carbon canister technologies  (U.S. EPA,   1980).   Such
systems are more effective, however, for regular  grade  gasoline containing ?5 to
27$  aromatics than  for  premium grade  unleaded gasoline containing H3$ aromatics
(U.S.  EPA,  1980).
     Most  of the  current diesel exhaust  emission studies  are  concerned with
emission  controls  through either engine design  or the  use of  fuel additives.
Other  control  options,  such  as  catalytic  reactors,  appear   to  be  viable
(Santodonato et al.,  1978).
     Other  major  sources  of automobile  emissions  are  losses  from  spilled
gasoline and losses during fuel transfer.  The former  can  be  reduced by educating
the  public  about the  necessity  of restricting  spillage both  for economic an».J
environmental reasons.   The loss of  gasoline during  fuel  transfer is already
controlled in many areas  of the country by incorporating vapor recovery systems
(NRC, 1980).
                                      5-1

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5.2.  ABATEMENT PRACTICES FOR SOLVENT USAGE
     Solvent vapors originating  from  industrial  use of toluene in coatings and
thinners can be controlled  or  recovered  by applying condensation, compression,
adsorption, or combustion principles.  Control efficiencies of 90$  or  greater are
possible by activated carbon adsorption, provided that particulates  are removed
from the contaminated  airstream by filtration before  the  airstream. enters the
carbon bed  (U.S. EPA, 1980).
     When  recovery  of  the vapor is not  desired,  an incineration method can be
used for controlling emissions.   The  choice  between direct flame and catalytic
incineration methods  must be  based  on  economic  factors  and  on local emission
standards.
     Control of toluene emissions  from gravure printing can be  done  in a number
of  ways  (U.S. EPA,  I960).  Process modifications  involving microwave, infrared,
electron beam,  or  ultraviolet  drying  and subsequent recovery of organic vapors
will reduce emissions.   Another alternative is to replace inks containing organic
solvents with aqueous or solventless inks.  Incineration of the  exhaust gases by
thermal or  catalytic methods provides  another method of emission control.  Last,
solvent  vapors  can be adsorbed  in  activated carbon as a method of  controlling
toluene  vapor emissions into the atmosphere.
5.3-   ABATEMENT FOR JOKE  OVEN  EMISSIONS
     Hydrocarbon  emissions result from the burning of the  stripped coke oven gas
for the  under-firing  of the  coke batteries.   The combustion exhaust gases from
each oven  are combined togecher and vented through a common stack.   Improving the
combustion efficiency  of  ~he coke  batteries  would be a proper method of control
 (U.S.  EPA,  1980).
5.1.    ABATEMENT  FOR EMISSIONS FROM MANUFACTURING SITES
     Current  technology for the control  of  gaseous hydrocarbon emissions from
manufacturing  sites  takes  the  form  of  charcoal  adsorption,   direct  flame or
catalytic   incineration,  chemical  sorbents, vapor  condensation,  process  and
material change,  and  improved  maintenance  (U.S.  EPA,  1980).  The  feasibility of
sorbing  organics  by the wet scrubbing method, using selected aqueous surfactant
systems  as opposed  to plain water, has been demonstrated  (Matunas  et  al.,  1978).
Organic  removal as high  as  90  to  95$  can  be  attained  by  using this method.
Condensation of organics  by the removal of heat may be an  expensive method since
refrigeration must  be used  for the removal of heat  from gases  (Matunar et al.,
1978).
                                       5-2

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5.5.  ABATEMENT PRACTICES FOR RAW AND FINISHED WATERS
     No information could be  found  on  this subject.   Treating water with acti-
vated carbon, howe;er, is expected to remove toluene from drinking waters.
5.6.  ECONOMIC BENEFITS OF CONTROLLING TOLUENE EMISSIONS
     There is no  significant  geographical area  in  the United  States  in which
ambient concentrations  of alkylbenzenes are  known  to be  harmful  to plants or
animal lives (NBC, 1980);  however,  as reactive hydrocarbons, they can contribute
to  the formation of photo-chemical smog that is known to be harmful to life and
property.  Brookshire et al.  (1979)  selected residential properties in six pairs
of  selected  neighborhoods and  found the  property value  could  increase on the
average of $504 annually if  the  air  quality were  improved.  The authors ascribed
about one-half of the enhanced  value to respondent-perceived aesthetic benefits
(visibility) and the other half  to perceived health benefits.  Thayer and Schulze
(1980) extrapolated the results of Brookshire et al. (1979) to the entire south
coast air basin of California  and concluded that  the urban benefits from improved
air quality  amounted  to between $1.6 billion  and $3  billion in the basin.  The
benefits that an improved air  quality would provide for commercial agriculture in
southern California  can  be  added  to the urban benefits described above.  Adams
et  al. (1980) examined the economic impact of ambient oxidants upon 14 selected
crops  in the region.   They  extrapolated their results of  these 14 crops to all
southern   California   commercial   agricultural  products  and   predicted  a
$250 million benefit  to be derived  from control  of oxidants in the air.
     All of  the  cost  benefits discussed above are based on total pollutants in
air.  It is  not  possible to  project the  portion of  these benefits that may be
attributable to  the  control  of  toluene  pollution  alone.    For  a  detailed
description   of   the   cost  benefits   of   controlling  alkylbenzene  'pollution,
interested  readers  are  referred to  a recent NRC  (1980) document.
5.7.  REFERENCES

ADAMS,  R.M.,  CROCKER,  T.D.  and  THANAVIBULCHAI,  N.    (1980).    An  Economic
Assessment   of  Air  Pollution  Damages  to  Selected  Annual  Crops  in  Southern
California.   U.S. Environmental Protection Agency, Washington, DC, 27 pp.
                                      5-3

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BROOKSHIRE, D.S., D'ARGE,  R.C.,  SCHULZE,  W.D.  and THAYER,  M.  (1979).  Methods
for valuing  aesthetics  and health  effects in the  south  coast air  basin:   An
overview.   Paper presented  at  the  72nd  Annual  Meeting  of  the  Air Pollution
Control Association, June 24-28, 1979, Cincinnati,  OH, 27 pp.

MATUNAR, F.C., TRATTNER,  R.B.  and CHEREMISINOFF,  P.N.  (1978).  The absorption of
organic compounds by wet scrubbing methods. Adv.  Instrum.  33.1 : 307-31^.

NRC  (NATIONAL RESEARCH  COUNCIL).   (1980).   The  Alkyl  Benzenes.   Committee on
Alkyl Benzene Derivatives, Board on  Toxicology and Environment Health Hazards.
Assembly of Life Sciences, National Research Council.  Washington, DC:  National
Academy Press.

SANTODONATO,  J., BASU, D., and  HOWARD, P.H.  (1978).  Health Effects Associated
with Diesel Exhaust Emissions, Literature Review and Evaluation.  EPA Publ.  No.
EPA-600/1-78-063,   Prepared for  U.S.  EPA,  HERL.,  NC.

THAYER, M. and  SCHULZE, W.D.   (1980).   An Examination of Benefits  and Costs of
Achieving  Ambient  Standards  in  the  South  Coast  Air Basin.   U.S. Environmental
Protection Agency,  Washington, DC.

U.S.  EPA (U.S.  ENVIRONMENTAL PROTECTION  AGENCY).   (1980).   Volatile Organic
Compound  (VOC)  Species Data  Manual,  2nd ed.,  Publication No.  EPA-A50/4-80-015.
Office  of  Air,  Noise,   and  Radiation,  Office  of Air  Quality Planning  and
Standards, Research Triangle  Park, NC.
                                      5-4

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              6.  ENVIRONMENTAL  FATE, TRANSPORT, AND  PERSISTENCE

     The environmental fate, transport, and  transformation of toluene in three
different media—air, water, and soil, are individually discussed below.
6.1.  AIR
6.T.I.  Fate in Air.  Toluene can persist in the atmosphere.  It is, therefore, a
prime candidate  for  short- and  long-range  transport  away  from urban emission
sources.  The dispersion  of toluene  from a point source to  the ambient atmosphere
can be modeled theoretically by  using dispersion equations.   One such modeling
method has been used in the Integrated Exposure Analysis Section (Section 10) to
determine the transport characteristics of toluene.
     The atmospheric toluene concentration downwind from one of the largest U.S.
automobile manufacturing plants was  measured by Sexton  and  Westburg (1980).  At a
point  6 km  from  the plar.t  site,   the  toluene concentration  was  found to  be
20.5 ppb.   The concentration  of  toluene  was  still 15.1  ppb  at a  point  18 km
downwind.
     Toluene itself-does not absorb  light  at wavelengths longer than  295  nm.  The
solar spectrum in the troposphere  does  not  contain  much  light of  wavelengths
shorter than 295 nm.  Therefore,  toluene can absorb only insignificant  amounts of
sunlight in  the lower atmosphere, but a charge-transfer complex between toluene
and molecular  oxygen absorbs light of wavelengths  to at least  350 nm.  According
to  Wei  and Adelman  (1969),  it is  the photolysis  of  this complex  that may be
responsible  for some of  the observed  photochemical  reactions of toluene.
      Toluene  apparently  is removed  from the atmosphere primarily  through  free
radical  chain processes  (NRC,  1980).  Of the  free radicals in the atmosphere,
hydroxy  (-OH),  atomic oxygen (0), and peroxy (*HO_  or  -R0?, whers R is an alkyl
or  acyl  group)  radicals  are potential initiators for the removal of  toluene.  An
additional  reactive  species is ozone.  The rate constants for tne  reaction of
these species  with  toluene and their relative  significance for toluene removal
are given  in Table 6-1.
      It  is  obvious  from  Table  6-1  that  reactions  with hydroxy  radicals are the
most important processes for the removal of toluene from the atmosphere.  Based
.upon  an  estimated daytime hydroxy  concentration given in Table 6-1 and a rate
constant for the reaction  of -OH radicals with toluene of  6.1  x 10"   cm^ mol"1
sec"  (Perry et  al.,  1977),  the  chemical  lifetime of toluene  in daylight hours
has been estimated to be  ^3 hours.  The atmospheric  residence time of toluene due
                                      6-1

-------
                                  TABLE 6-1
                Rate  Constants  for Reactions  of Toluene with
                    Reactive Species in the Atnosphere
Estimated Average Rate of
Daytime Annual Toluene
Concentration
Species ppcn
Hydroxyl „
radical 1 x 10"°
Atomic
oxygen 3 x 10~
Peroxy _^
radical 1 x 10
Ozone 3 x 10
Rate Constant, Removal, Fraction of
ppm min ppm/min Hydroxyl Rate
9.5 x 103 3.? x 10"^ 1
1.1 x 102 3.3 x 10~7 10~3
2.5 x 10~7 2,5 x 10~11 4 x 1C"8
5 x 10~7 1.5 x 10""8 5 x 10~5
Source:  NRC, 1980
                                     6-2

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to "OH radical  reactions  has been estimated  to  be 1.9 days  by Cupitt (1980),
However,   the  half-life  or  residence  time  value  is  subject  to considerable
uncertainty and may  vary on  a  day-to-day  basis  by  as  much  as an  order of
magnitude  depending  on  sola'r   intensity,  temperature,  and  local   trace  gas
composition of the atmosphere.
     The  reaction  products  formed  from  toluene under  simulated atmospheric
conditions  are  not known  with  certainty.   According  to  the study  of O'Brien
et al.  '(1979), the  gaseous  products  of the reaction  are  o-cresol,  m- and p_-
nitrotolaene,  benzyl nitrate, and benzaldehyde.   Of these  products, o-cresol and
benzaldehyde  are the major  components,  each  composing about  8%  of  the total
product yield.   The mechanisms by  wh.Joh  these products are formed are shown in
Figure 6-1.
     It is assumed that the reaction proceeds  via addition of «OH radicals to the
ring or by abstraction of hydrogen from the methyl side  chain.   Several investi-
gators have determined  the relative  importance of both  reaction pathways.   From
the  amounts of reaction  products  formed,  it was  determined  that the addition
mechanism is of much greater significance than the abstraction mechanism  (Kenley
et al., 1978; O'Brien et  al.,  1979;  Hoshino et al., 1978).
     Other  reaction  products also  are  formed  from   toluene  reactions under
simulated atmospheric  conditions.   Some  of  the  ring  fragmentation products
formed  are acetylene,  acetaldehyde,  and  acetone.   The  total  yield of these
products  is much less than 1J.  Formaldehyde and formic acid are-also formed, but
their  yields  are not  known.   A measurement  of the total gas phase carbon  showed
that 60%  of the oxidation products from the  photodecomposition  of toluene  left
the  gas  phase  and  deposited  on  the walls  of the reaction vessel or  formed an
aerosol  (NRC, 1980).  The distribution of the products  between gas and condensed
phases  (aerosol) in the open atmosphere  (as opposed to the reaction in a  vessel)
is still  not  clear.
      In  addition to the above  photooxidation  products,  photolysis of  toluene  in
polluted  atmospheres (containing NO ) yields ozone and fairly  high  amounts  of
peroxyacetylnitrate  (PAN) (5  to 30$  nitrogen  yield)  and  peroxybenzoylnitrate
(PBzN)  (0 to  5% nitrogen  yield)  (NRC,  1980).   The mechanism of  PAN formation  is
either  by the fragmentation  of  the aromatic  ring or by the secondary reactions
involving products of toluene  photolysis.   PBzN  is formed by  the photooxidation
of benzaldehyde produced  from the  photooxidation of toluene (NRC, 1980).   The
formation of  the  peroxy   compounds  is significant because these products  are
                                       6-3

-------
CH,
               .OH
                       addition
         OH
             •f   HO,
CH,
                                                                   CH,
                                                OH
                                                                         OH
                                                NO.
                                          CH,
                                                              CH.
                                                OH
                                                NO,
CH-
                                                                      CH,
             .OH
                               FIGURE 6-1




  Proposed  Reaction Pathways of Toluene Under Atmospheric Conditions




                          Source:   NRC, 1980
                                6-H

-------
strong eye irritants  tind oxidizing agents,  and may induce  plant  damage (NRG,
I960). For  an  excellent  review of  the photochemical  fate  of  toluene  in the
atmosphere, the reader is referred to a recent NRC document  (NRC, 1980).
6.1.2.   Transport.   The volatility of  toluene  and  its low solubility in water
permit it  to  volatilize from  water  surfaces  to the  atmosphere  (MacKay  and
Wolkoff,  1973).  Studies of actual  and simulated oil  spills in seawater indicate
that virtually all hydrocarbons smaller than C1t- will be lost to the atmosphere
within a few days (McAuliffe, 1977).  The reverse process, that is, transfer of
toluene  from air to hydrosphere through rain,  is also  known to occur (Walker,
1976); however,  washout should be considered an insignificant  removal process
for toluene from air (NRC, 1980).  The  probability of  physical removal of toluene
from the atmosphere was also speculated to be unlikely by  Cupitt (1980).
6.2.  AQUATIC MEDIA
6.2;1.   Fate.   Sauer  et al. (1978) concluded from their studies of the coastal
waters of the Gulf of Mexico that toluene and other alkylbenzenes are persistent
in "the marine environment.  The probable modes of toluene loss or transformation
from the aquatic environment are discussed below.
     Oxidation:   Reaction of toluene in water  with  hydroxy  radicals generated
from the irradiation of hydrogen peroxide produces benzaldehyde,  benzyl alcohol,
and cresols (Jefcoate et al., 1969).  No data were found in  the literature from
which a relevant rate  of oxidation of toluene in  the aquatic environment could be
detenu ned.
     It  has  been observed  (Carlson  et  al., 1975) that  toluene may form small
amounts  of  chlorine-substituted products during chlorination  under conditions
used  for water  renovation.    The  extent  of chlorination  increases  with  the
decrease of pH and increase  of co tact tire.   At a water temperature of 25°C and a
                                 h
chlorine concentration of 7 x 10   M,  the percent chlorine  uptake was determined
to be 11.1 and 2.9$  at water pH  of  3 and 7, respectively (Carlson et al., 1975).
With  ether  conditions remaining the  same,  no  chlorine  uptake  was observed at
water pH of 10.1.  Therefore,  chlorination  of renovated water which is usually
carried out at pH levels near 7  may not  be of significant environmental concern.
     Hydrolysis:   No  data  have  been  found that  would  support  any  role  of
hydrolysis in the fate of toluene in  the aquatic medium.
     Bioaccumulation:  No measured  steady-state  bioconcentration factor  (BCF) is
available for  toluene but,  using  the equation  of Veith et  al. (1979)  and the
measured  octanol-water   partition  coefficient  of 2.51  (U.S.  EPA,  1980)  (as
opposed to the theoretical  value for  log BCF of  2.69  [Chiou  et  al., 1977]), the

                                      6-5

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U.S. EPA (1930) has estimated the BCF as 27.1.  A  factor of 3-0/7.6  = 0.395 has
been used  to  adjust the estimated  BCF  from the 7.6$ lipids on which the Veith
et al. (1979)  equation is based to the 3? lipids th?.t is the weighted  average for
consumed fish and shellfish in the United States.   Thus, the weighted  average BCF
for  toluene  from  edible  aquatic  organisms  consumed by  Americans has  been
calculated to be  27.1 x 0.395 =  10.7.
     In one experiment (Roubal et al.,  1978), coho  salmon (Oncorhynchus kisutch)
and starry flounder (Platichthys  stellatus)  were exposed to  a soluble  fraction of
a crude oil containing aromatic hydrocarbon  in a flowing seawater.  It was found
that alkylated aromatics accumulated in  tissues to a  greater degree  than unsub-
stituted  derivatives.   In both  species,  accumulations of  substituted benzenes
increased  with increased alkylation.  The tissues  were net analyzed  for toluene
because of inadequate  analytical procedures.  It  was  determined, however, that
the  bioconeentration  factors in  starry flounder for  Cj,  and  Cr  substituted
bea-zenes  were as high as  2600  and as low  as  near zero (concentration  in fish
tissue  was  below  detection  limit of  0.05  ppm)  for  xylenes.     Substantial
variations  in BCF for  individual hydrocarbons were found in both species.  The
muscle of  coho  salmon,  which has a higher  lipid  content  than starry flounder,
showed .a  lower  BCF.  It was concluded (Roubal et  al.,  1978) that factors other
than lipid content were more  important in the observed species  differences in the
BCF values.
6.2.2.  Transport.  The primary fate-determining processes  of  toluene in aqueous
media appear  to  be its intermedia  transport processes (U.S.   EPA,  1979).   The
details of the transport processes  are discussed below.
     Water to Air:   Although there  are no experimentally determined  evaporation
rates  ot  toluene  from  water,   there  are  theoretical models available  for
predicting the  rate of evaporation of  slightly-soluble materials  from aqueous
solution  (Mackay  and Wolkoff,  1973; Liss and Slater,  1974; Mackay and Leinonen,
1975; Dilling,  1977).   The most  accurate of these  is  based on  the mass transfer
coefficients  for  the liquid  and  vapor phases reported by Liss  and Slater (197*0
and the Henry's law constant for a solute as calculated by  its  solubility, vapor
pressure,  and rnclecular weight  (Mackay  and Leinonen,  1975).   Based on these,
Mackay  and Leinonen (1975)  reported  the calculated  evaporation  half-life for
toluene from  1m  deep water to be 5.18 hours.
     The  intramedla transfer of  toluene  in water  can  be  calculated from this
half-life  (t  1/2) value.   If the  t  .  and the current  Telocity are  assumed to be
                                       6-6

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5.18 hours and  1  m/sec,  respectively,  the distance downstream  that  water in a
river would flow before the volatilization of 50% toluene is:
                 5.18 hour  x  1  m/sec  x  3600 sec/hour = 18,648 m
     Similarly,  Henry's  law  coefficient  (H)  can be  used  to determine toluene
concentration in  air  phase over seawater.  If the height of the  air and water
columns are assumed to be the same, the  Henry's law coefficient can be given as:
                    [toluene ]
               H = -FT—,	r5^  = 0.3^9 for seawater  (NRC, 1980)
                    [toluene ]liq

Thus, if equilibrium were  attained,  only  26% of  toluene would be present in the
gas phase above seawater.  This  calculation does not  consider stratification.
     In natural shallow or deep waters where stratification is expected  to occur,
it is likely that the atmospheric mixing layer is 10 to 100 times deeper than the
aquatic mixing layer (NRC, 1980).  In such water, 78 or 97?, respectively, of the
toluene would exist in the gas  phase.
     Water to Soil:  The importance of  this transport  process can be evaluated by
experimentally  determining the  toluene content  in sediments  of  surface water
contaminated  with  toluene.    Theoretical modeling can  also be  used  for this
purpose.   Using  the U.S.  EPA's  multicompartment  Exposure  Analysis  Modeling
;5ystem  (EXAMS),  Slimak  (1980)  has determined that bottom sedimonts account for
ever 90$ of the  total toluene  discharged  into surface waters under steady-state
conditions.  The values for the distribution of toluene between surface  water and
sudiment as  determined by  the.EXAMS  modeling do  not agree with the experimental
results  of Jungclaus  et al.  (1978).   Jungclaus et  al.  (1978)  determined the
toluene  content  in  the  water  and  sediments of a river  receiving wastewater
containing  toluene.   Although toluene  was  detected  in the river water, it was
found not to accumulate  in the sediments. More  research  in  this area  is needed
to  explain this  discrepancy  between the EXAMS modeling  and  the experimental
results.
6.3.  SOIL
6.3.1.   Fate.    Toluene  probably exists in soils in  the  sorbed  state.   The
sorption of  toluene by clay minerals  (bentonite and  kaolinite)  was studied by
El-Dib et al.   (1978) and  was  found  to  follow Freundlich's adsorption  isotherm.
These authors also  found  that  the  adsorption capacity increased as the pH  value
decreased.
     The fate of  toluene in soil has not been thoroughly investigated.  It can be
anticipated, however, that a portion of toluene  in  soil will undergo intermedia
                                       6-7

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transfer to air and water, and a portion will undergo intramwdia transfer.  The
part that stays in soil may participate in chemical reactions (including photo-
chemical reactions) and biological degradation and transformation.  The relative
importance  of  intermedia  transfer  and  chemical  and  biological  reactions  of
toluene in soils is not known.
     Investigations by  Wilson  et  al. (1981)  indicate that volatilization, bio-
degradation,  and  biotransfonnation  processes  dominate  the fate of  toluene  in
'soils.  The intermedia  transfer of toluene from soil to  water  probably is not an
important pathway.  No data could  be  found in  the  literature searched that would
support a hypothesis  for any role of chemical reactions in determining the fate
of toluene in soils.   The intermedia transport  of toluene and its biological fate
in soils have been discussed separately below.
6.3.2.  Transport.
     6.3.2.1.  SOIL TO AIR — Laboratory experiments of Wilson et al.  (1981) show
that 38 to 66$ of 0.2  to 0.9 mg/& of toluene applied to the  surface of  sandy soils
with 0.087?  organic carbon will volatilize  to air.  The volatilization rate is
dependent on  the  nature of the soil.  The volatilization  rate  may be signifi-
cantly  lower  for  soils  with  high  organic   contents  due  to   their  sorption
properties  (Slimak,  1980).   This  phenomenon  may  be especially important with
respect to municipal  sludges  that normally contain high organic substances.
     6.3.2.2.   SOIL TO  WATER  — The  transfer  of toluene from soil  to ground or
surface waters  can be of importance  with regard  to the possibility of contami-
nation  of  these water  bodies and  their  subsequent use as  sources of drinking
waters.  Unfortunately, very little information is available about  this subject.
From the investigations of Wilson  et al.   (1981). it can  be concluded that the
transport of toluene from soil to  water is probably not a major transfer pathway.
These investigators showed that 2 to 13? of the applied toluene on a sandy soil
system  could  be eluted through a  column  140  cm high.   The leaching of toluene
from landfill sites that contain  soil originated  partly from municipal sludges
can be expected to be even lower.  The higher  organic content  of these soils may
"etard the aqueou? elution  process due to higher sorption properties of the soils
toward toluene.
6.4.  ENVIRONMENTAL PERSISTENCE
6.4.1.  Biodegradation  and Biotransfonnation
     6.4.1.1.   MIXED  CULTURES — The  study of the disappearance of toluene in
soil began  nearly  75  years ago.   Stormer (1908)  and Wagner  (1914) showed that
                                       6-8

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toluene was  susceptible  to  bacterial  decomposition in  the soil.   Gray  and
Thornton (1928) and Tausson (1929) isolated soil bacteria  that used toluene as a
sole carbon source.  Glaus and Walker  (1961) found that the half-life of toluene
in isolated bacteria from soil inhabited with toluene-degrading bacteria was 20
to 60 minutes.  Wilson et al.  (1981) indicated that from 21 to 60? of toluene
eluted through  1*40  cm  of sandy soil  biodegraded.   The  authors  stated that the
process was probably very sensitive to the soil typ-? and, therefore, may or may
not be an important removal process of toluene  from  a particular soil system.
     More  literature,  however,  exists   on the biodegradation  of  toluene  in
aquatic environments.  In a U.S. EPA report (Slimak,  1980), the biodegradation of
toluene in lakes,  rivers, and ponds was  discussed using an extension of the U.S.
Environmental Protection Agency's (U.S.  EPA) multicompartment Exposure Analysis
Modeling System (EXAMS).   The report  stated that the biodegradation of toluene
accounted for  0.31,  4.81,  0.36,  0.09, and  18.47? of the  total  toluene loss in
oligotrophic  lakes,  eutrophic  lakes, clean  rivers,  turbid  rivers,  and  ponds,
respectively.   Using  the  standard  dilution  method  and  filtered  wastewater
effluent  as  the seed  to determine  the biochemical  oxygen der^nd  (BOD),  the
biodegradability  (percentage bio-oxidized)  of toluene ranged  from 63  to  86?
after up to 20 days (Price et al., 1974; Bridie et al.,  1979).
     Matsui et al.  (1975)  found that  in activated sludge  acclimated to various
organic compounds, the total  organic  carbon (TOC) removal  efficiency for toluene
was 60? while the  chemical oxygen  demand (COD) was 72? for 24 hours.  The authors
concluded, however, that although toluene was a readily biodegradable compound,
in  this  experiment  disappearance was  due mainly  to  evaporation.    Using  the
Warburg technique, Lutin et al.  (1965)  reported a MO? degradation of toluene in
activated sludge after 144 hours.   In  comparison, 63? of the benzene was degraded
in the same  time.   The  degradation of  toluene in benzene-acclimated activated
sludge reacned 17.2? of the theoretical  BOD after 6 hours and 48? after 192 hours
(Malaney and McKinney,  1966).  Toluene was  the most biodegradable of a number of
alkylbenzenes tested by these authors, who also found that the introduction of a
methyl group  to benzene retarded the  initial (6 hour)  rate  of oxidation  of
toluene  but  not  the  extent  of  degradation compared to  benzene.    Marion  and
Malaney (1964) exposed activated sludge to 500  mg/S,  of toluene from 3 municipal
plants and reported that unacclimated sludge showed little ability to oxidize
benzene  and   toluene  after  6  hours   and  that  after 72 hours,   less  than  11?
oxidation had taken place  (compared  to  44.7?  reported by Malaney and McKinney,
                                      6-9

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1966).   One sludge sample,  however, acclimated to benzene, oxidized greater than
30$ of  the  toluene after  180 hpurs.   It  should  be  noted that the high concen-
tration of toluene used in  this  study probably was toxic to the organisms in the
sludge.
     Tabak et al-. (1981) studied the biodegradation of toluene at lower concen-
trations  (5 mg/S,  and  10 mg/&)  with  settled  domestic wastewater  as microbial
inoculum  by  the   static-culture   flask-screening   procedure.     Toluene  was
completely biodegraded in  7 days at 25°C  by  this method.   At influent  concen-
tration  levels  of  70  \ig/i and  ">12 ng/£,  the  removal of  toluene by activated
sludge  treatment  at  two municipal  facilities  was  found  to  be at  least  99?
(Patterson and Kodukala, 1981).   In  31  industrial  facilities, activated sludge
treatment was found to remove an average  of  52?  toluene  at an average influent
concentration of  119  ng/&.   Tne  treatment  of  three industrial  effluents  in
aerated  lagoons  removed an  average  of  >93? toluene  initially   present  at  an
average concentration of 143 (J.g/& (Patterson and Koaukala, 1981).
      i
     The  degradation  of toluene  has also  been  studied  in  mixed  cultures  of
bacteria.  Chambers et al. (.1963),  using phenol-adapted  bacteria, reported 38?
degradation of toluene after 180 minutes.   It should be noted that phenol is the
metabolic degradation pathway of toluene.   In another study, Dechev and Damyanova
(1977)  grew  sludge cultures in either phenol, xylene,  or toluene  as  the sole
carbon source and found that phenol-adapted bacceria proved less  able to degrade
xylene  and  toluene,  while  toluene-adapted cells showed  greater  versatility  in
their ability to oxidize phenol and xylene.
     6.4.1.2.  PURE CULTURES — Although pure cultures do not occur  in nature, a
discussion of biodegradation in  these media may provide insight to  degradation in
more complex media occurring in a natural  environment.  Fungi and bacteria have
been shown to use toluene  (Smith and Rosazza, 1974).  In the course of studying
the  effects  of  toluene  on microbial  activity,  Kaplan  and  Hartenstein (1979)
discovered that  6 of 7  fungi .Imperfecti,  7  of 13 basidiomycetes,  and  6  of  14
bacteria grew with 0.1 or  0.05* toluene as the sole  carbon  source.  The addition
of yeast extract  increased the  amount  of toluene-utilizing microorganisms.   In
contrast, no oil-use or hydrocarbon-degrading fungi grew on  toluene as the sole
carbon source (Davies and Wcstlake,  1979). Using an oxygen electrode to measure
oxidation, Buswell and Jurtshuk (1559) found that resting cells  of an n-octane-
utilizing Corynebacterium sp. oxidized only 7% of the  available toluene compared
to 100? oxidation  of  n-octane.   Toluene  did not  serve as a  growth substrate in
                                      6-10

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this experiment.  Kapraleck (1954) isolated a Pseudomonas-type  bacteria  from the
soil of a petroleum deposit that  used toluene.  Pseudomonas sp.  and Achromobacter
sp.   from soil used  toluene  as  the sole  carbon source  for  growth  (Glaus and
Walker,  1964;  Gibson and  Yeh,  1973).   Smith  and  Rosazza (1974)  reported that
bacteria and yeast hydhoxylated  toluene.   In contrast, Nei et  al.  (1973) found
little oxidation of toluene by phenol-using  yeast.
     The  metabolic pathway  for  the  bacterial  oxidation  of  toluene  has been
studied with soil  microorganisms  (Figure 6-2)  and reviewed by  Gibson (1971) and
Subramani-an et  al.   (1978).   On the basis of simultaneous adaptation  studies,
Kitagawa  (1956)  concluded that  Pseudomonas  aeruginosa  oxidized toluene  via
benzyl  alcohol  and benzaldehyde  to' benzoic acid and  ther to  catechol.   This
pathway was  supported by the investigations  of Nozaka and Kusunose  (1968).  A
Mycobacterium sp.  also  produced  benzoic acid from toluene (Atkinson  and Newth,
1968),  as did  a methanotrophic bacterium  (Methylosinus trichosporium)  (Higgins
et al., 1980).
     An  alternative pathway  was proposed  by  Glaus and  Walker  (1964)  using a
Pseudomonas sp. and an Achromobacter sp. isolated from soil that used toluene as
a  sole carbon  source  for  growth.  These  investigators  found that washed cell
suspensions  oxidized toluene to 3-methylcatechol,  indicating  that  the methyl
moiety was not  oxidized, as occurred in the pathway proposed  by Kitagawa (1956).
A . similar oxidation  product was  found by  Nozaka  and  Kusunose  (1969) using
Pseudomonas  mildenbergii  cell-free  extracts.   Gibson   et  al.   (1968a)  also
reported  the detection of  3-methylcatechol from toluene by Pseudomonas  putida.
An oxidation product preceding 3-methylcatechcl was  found in cultures  of a mutant
strain of P_.  putida (strain 39/D)  (Gibson £t al., 1968b, 1970).   This new product
was  identified as  (+)-cis-2,3-dihydroxy-1-methylcyclohexa-4,6-diene  ( cis-2,3-
dihydro-2,3-dihydroxytoluene)  (  c^s-2,3-DH-2,3-DOH  TOL)  (Kobal et  al.,  1973).
The catechol and  3-methylcatechol then can be cleaved by  ortho cleavage  to yield
the corresponding muconic  acids  or  by meta cleavage to "ield  the corresponding
hydroxymuconic  semialdehydes  (Bayly  et  al.,   1966).   Methylmuconic  acid was
formed  from  toluene oxidation by a soil  bacterium Nocardia corallina  (Jamison
et al.,  1969).   The  semialdehydes  are further converted to  2-hydroxy-6-oxo-
2,cis-4,cis-heptadienoic acid (2-OH-6-0X0-2,cis-4, cis-HA) and then to  acetate,
pyruvate,  and  acetalydehyde  and to  C0_ and  energy (Bayly et al.,  1966).   The
conversion of  toluene to  compounds that  can be used  as  sources  of  carbon  and
                                      6-11

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                          CH,
                       TOLUENE
       • fNZYl 4LCOHOL
                               X
                                tB-J. 1-DM-I J-DOH TOL
        »ENZ»LD£KroE
         • ENZOICtCID
                 \
                                  J-MtTHTLCATtCMOL
                                    /
                                  /

                                 '
                          fttETHVUWUCONIC
                              ACID
IHUCON'C ACtO
                HVDBOKVMUCONIC
                 ttMIALRfHVDt
               	J
MrTHYLNYDROSYMDCOWIC
    HMIALDtMVOt
                                            rtTALDEHVDE
                                            ACtlATE
                                          CO, • tutncv
                          FIGURE 6-2

            Microbial  Metabolism  of  Toluene

                              6-12

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                              I
energy suggests that toluene will be degraded rapidly by these microbiai species
proliferating at the expense of  the compound.
     The enzymes  responsible for  toluene  degradation are  carried  on plasmids
(Williams and Worsey, 1976;  Saunders,  1977).  Williams and  Worsey (1976) isolated
13  bacteria  from soil,  all of  which carried  the  toluene-degrading plasmids,
suggesting that  the plasmid-borne  gene  responsible  for  toluene degradation is
wide spread in the soil microbiai population.  The plasmid  can also be  transposed
into  other  hosts,  further  increasing the  number of toluene-degrading bacteria
(Broad et al., 1977; Jacoby et  al.,  1978).   The toluene plasmid in Pseudomonas
putida coded for the  metabolism  of  toluene to  the corresponding  alcohol and
aldehyde via the meta pathway, to  the semialdehyde and further  products (Worsey
and Williams,  1975;  Worsey  et^ a_l.,  1978).  A plasmid coding for  both toluene and
xylene degradation  in a  Pseudomonas sp.  has been isolated recently and charac-
terized  (Yano  and  Nishi, 1980).   Broad  et  al.  (1977) have speculated that the
ortho pathway  of toluene  degradation  probably is  chromosomally  coded.

6.5.  REFERENCES

ATKINSON, J.H. andF.H.  NEWTH.   (19&8).  Microbiological transformation of hydro-
carbons.  Proc. Microbiol.  Conf. p. 35-^5.

BAYLY, R.C. et al.   (1966).   The metabolism  of cresols by species of Pseudomonas.
Biochem. _J.   101 : 293-301.

BRIDIE,  A.L.  et  al.   (1979).    BOD  and  COD  of some petrochemicals.   Water
Research.  t3: 627-630.

BROAD, P.,  BAYLEY,  S.,  DUGGLEBY, C.J., WORSEY,  M.J. and WILLIAM, P.A.  (1977).
Plasmid  degradation  of toluene and xylenes in soil pseudomonads. In:  Plasmids.
Medical  and  Theoretical  Aspects.  Mitsuhashi, S., Rosival, L.  and Krcmery, V.,
eds.  Berlin:  Springer-Verlag  KG., p. H03-405.

BUSWELL,  J.A.  and  JURTSHUK,  P.    (1969).   Microbiai oxidation of hydrocarbons
measure  by oxygraphy.  Appl. Mikrobiol.   64_:  215-222.
                                      6-13

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CARLSON,   R.M.,  CARLSON,  R.E.,  KOPPERMAN, H.L.  and CAPLE,  R.    1975.   Facile
incorporation of chlorine  into aromatic  systems  during  aqueous  chlorination
processes.  Environ. Sci. Technol.  £(7): 674-675.

CHAMBERS, C.W.  et al.    (1963).   Degradation  of aromatic compounds by  phenol-
adapted bacteria.  J. Water Pollut. Cent. Fed.   35(12):  1517-1528.

CHIOU, C.T., FREED, V.H., SCHMEDDING, D.W. and KOHNERT,  R.L.   (1977).   Partition
coefficient and  bioaccuraulation of selected  organic  chemicals.   Environ.  Sci.
Technol.  11(5):  475-578.

GLAUS, D. and WALKER,  N.  (1964).  The decomposition of toluene by soil  bacteria.
J.  Gen. Microbiol.  36_:  107-122.

CUPITT,  L.T.     (1980).    Fate  of  Toxic and Hazardous  Materials  in  the Air
Environment.    U.S. EPA  Report  No.   600/53-80-084.     Environmental   Sciences
Research Laboratory, U.S.  EPA, Research Triangle Park, NC.  Available from  NTIS.
Order  No. PB 80-221948.   Springfield, VA.

DAVIES,  J.S. and WESTLAKE, D.W.S.  (1979).  Crude oil utilization by fungi.  Can.
J.  Microbiol.   25(2):  146-156.

DECHEV,  G.D. and  DAMYANOVA, A.A.  (1977).  Functional  investigation  of  bacterial
composition of active sludge.  Bulgarska Akademiiana Naukite.  Doklaly Bolgarskoi
Akndemiia Nauk.   30(10): 1475-1478.

DILLING, W.L.   (1977).   Interphase  transfer  processes.  II.  Evaporation  rates  of
chloro methanes,  ethanes,  ethylene,  proanes,  and propylenes  from dilute aqueous
theoretical predictions.  Environ.  Sci.  Technol.  JJ_(4):  405-409.

EL-DIB,  M.A.  et al.   (1978).   Role of  adsorbents in  the  removal of  soluble
aromatic  hydrocarbons  from  drinking  water.    Water Res.   12: 1131.   (Cited  in
Syracuse Research Corporation,  1980).
                                      6-14

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GRAY, P.H.H. and THORNTON,  H.C.   (1928).   Soil bacteria that decompose cert?in
aromatic compounds.  Abl. Bakt. (Abst. 2).  73: 74.  (Cited in Claus  and Walker,
1964).

GIBSON,  D.T.,  KOCH, J.R.,  SCHULD, C.L.-and  KALIO, R. E.   (1968a).    Oxidative
degradation of aromatic  hydrocarbons  by microorganisms.  I. Enzymatic  formation
of catechal from benzene.   Biochem.   7(7): 2653.

GIBSON,  D.T.,  KOCH, J.R.,  SCHULD,  C.L. and.  KALIO,  R.E.   (1968b).    Oxidative
degradation  of  aromatic  hydrocarbons by  microorganisms.  II.  Metabolism  of
halogenated aromatic hydrocarbons.  Biochem.   7(11): 3795-3802.

GIBSON,  D.T.,  HENSLEY,  M.,  YOSHIOKA,  H. and MABRY,  T.J.   (1970).    Oxidative
degradation  of  aromatic  hydrocarbons by  microorganisms.  III.   Formation  of
(+)-cis-2.3-dihydroxy-1-m-ethyl-4,6-cyclohexadiene from toluene  by  Pseudomonas
putida.  Biochem..   9(7):  1626-1630.

GIBSON,  D.T.  (1971).  Microbial oxidation of aromatic  hydrocarbons.  Crit.   rev.
Microbiol.  1(2):  1Q9-223-

GIBSON,  D.T.  and  YEH,  W.K.    (1973).    Microbial  degradation  of aromatic
hydrocarbons.  Microbiol.  Degradation Oil Poll.,  Workshop,  p. 33-38.

HIGGINS,  I.J.  et  al.    (1980).   New  findings  in methane-utilizing bacteria
highlight  their importance in the  biosphere and  their commercial  potential.
Nature.   286: 561.

KOSHINO, M.,  AKIMOTO,  H.  and  OKUDA,  M.   (1978).   Photochemical oxidation  of
benzene, toluene,   and  ethylbenzene  initiated by  hydroxyl  radicals  in the  gas
phase.   Bull.   Chem.  Soc.  Japan.    5J_: 718-724.   Taken  from:   Chera.  Abstr.
88:  I69346v,  1978.
                                      6-15

-------
JACGBY,  G.A.,  ROGERS,  J.E.,  JACOB,  A.E.  and HEDGES,  R.W.   (1978).  Transposition
of  Pseudomonas  toluene-degrading  genes  and  expression in  Eschei-ichia  coli.
Nature.   27^(5667): 179-180.

JAMISON, V.W.,  RAYMOND,  R.L.  and HUDSON,  J.O.   (1969).  Microbial  hydrocarbon
cooxidation. III. Isolaton and characterization  of  an  alpha. , .alpha. '-dimethyl-
cis.cis-muconic acid-producing strain of Nocardia cornallina.   Appl. Microbiol.
17(6): 853-856.

JEFCOATE,  C.R.E. et  al.  (1969).    Oxidation  of  some  benzenoid  compounds  by
Fenton's reagent and the ultraviolet irradiation by hydrogen peroxide.  ^J.  Chem.
Soc.  B. : 1013.  (Cited in Syracuse  Research Corporation,  1980).

JUNGCLAUS, G.A., LOPEZ-AVILA, V. and HITES,  R.A.   (1978).   Organic compounds  in
an industrial wastewater:  A case study of their environmental impact.  Environ.
Sci.  Technol.   12(1) : 88-96.

KAPLAN, D.L. and HARTENSTEIN, R.  (1979).   Problems with toluene  and the  deter-
mination  of  extracellular  enzyme  activity  in  soils.    Soil  Biol.  Biochem.
      : 335-338.
KAPRALEK,  F.    (195*0.     Assimilation   of   hydrocarbons   by  microorganisms.
Ceskoslcv.  Biol., 3_: 82-91  as  cited  in Walker,  ,976.

KENLEY, R.A. , DAVENPORT,  J.E. and  HENDRY,  D.G.   (1978).   Hydroxyl  radical  reac-
tions  in  the  gas phaase.   Products  and  pathways for the  reaction of OH  with
toluene.  _J.  Phys. Chem.   82_:  1095-1096.

KITAGAWA, M.  (1956).   Studies  on  the oxidation mechanism of methyl group.   J_.
Biochem.  H3(*Q : 553-563.

KOBAL, V.M., GIBSON, D.T., DAVIS,  R.E. and GARZA, A.  1973-   X-ray  determination
of  the absolute stereochemistry  of the initial  oxidation  product formed  from
toluene by Pseudomonas  putida 39/D.   _J. Amer.  Chem.  Soc.  95(13) :
                                      6-16

-------
LISS, P.S. and SLATER, p.G.  (1974).  Flux of gases across the air-sea interface.
Nature.  247: 181-184.

LUTIN,  P.A.,  CIBULKA,  J.J. and  MALANEY, G.W,   (1965).   Oxidation of  selected
carcinogenic compounds by activated sludge.   Purdue Univ.,  Eng,  Bull,  Ext.  Ser.
118: 131-115.

MACKAY, D.  and  WOLKOFF,  A.Q.   (1973).   Rate pf evaporation of  low-solubility
contaminants from water  bodies  to atmosphere.  Environ. Sci. Technol.  7_: 611.
(Cited  in Syracuse Research Corporation,  1980).

MACKAY, D.  and  LEINONEN,  P.J.   (1975).   Rate of evaporation of  low-solubility
contaminants  from   water  bodies  to  atmosphere.    Environ.   Sci.   Technol.
9:  1178-1180.

MALANEY, G.W. and MCKINNEY, R.E.  (1966).  Oxidative abilities of  benzene-accli-
mated activated sludge.   Water Sewage  Works.   113(8):  302-309.
     [
MARION,  C.V.  and  MALANEY,  G.W.     (1964).     Ability  of  activated -sludge
microorganisms to oxidize aromatic  organic  compounds.  Proc.  Indus. Waste Conf.
18:  297-308.

MATSUI, S. et al.  (1975).  Activated sludge degradability of organic  substances
in  the waste water of the Kashima petroleum  and petrochemical industrial complex
in  Japan.   Prog. Water  Technol.   ]_:  645-649.

MCAULIFFE, C.D.   (1-977).   Evaporation and solution of C- to C... hydrocarbons from
crude oils  on the sea surface.  Wolfe,  D.A.,  ed.   Fate and Effects of Petroleum
Hydrocarbons  in  Marine  Organisms and  Ecosystems.   New  York:  Pergamon Press,
p.  368-372.   (Cited in  Syracuse Research Corporation,  1980).
                                      6-17

-------
NEI,  N., ENATSU,  T.  and TERUI, G.   (1973).   Microbiological decomposition  of
phenols.  TV. Oxidation of aromatic compounds  by  pheno-utilizing  yeasts.   Hakko
Kogaku Zasshi.  51:1-11.  Taken from:  Chem. Abst.  7_8_:  9^6392, 1973.

NOZAKA,  J.  and  KUSUNOSE,   M.     (1968).    Metabolism  of  hydrocarbons   in
microorganisms.   I.  Oxidation  of p-xylene  and  toluene  by  cell-free  enzyme
preparations of Pseudomonas aeruginosa.   Agr.  Biol. Chem.  32(8)-.  1033-1039.

NOZAKA,  J.  and  KUSUNOSE,   M.     (1969).  ,  Metabolism  of  hydrocarbons   in
microorganisms.  II.  Degradation of toluene by  cell-free extracts  of Pseudomonas
mildenbergii.  Agr. Biol.  Chem.  _32t8):  1033-1039.

NRC  (NATIONAL  RESEARCH COUNCIL).   (1980).  The  Alkyl Benzenes.    Committee  on
AJkyl Benzene Derivatives,  Board on Toxicology  and Environmental Health  Hazards.
Assembly of Life Sciences,  National Research Council.   Washington,  DC.   National
Academy Press.

O'BRIEN, J.R.  et  al.   (1979).   Interaction of oxides of nitrogen  and  aromatic
hydrocarbons under simulated  atmospheric conditions.   Caphter 11,  in Grosjean,
D.,  ed.  Nitrogenous  Air Pollutants;  Chemical and Biological  Implications, Ann
Arbor,  MI:    Ann  Arbor Science  Publishers.   p.  189-220.    (Cited  in  Syracuse
Research Corporation,.  1980).

PATTERSON, J.W. and KODUKALA, P.S.  (1981).  Biodegradation of hazardous organic
pollutants.   CEP.  77CQ: 48-55.

PERRY,  R.A., ATKINSON, R. and PITTS, J.N., JR.   (1977). Kinetics and mechnism  of
the  gas phase  reaction of  OH radicals with  aromatic hydrocarbons  over the
temperature  range 296-473°K.   J.  Phys. Chem.   8l: 296-304.

PRICE,  K.S.,  WAGGY,  G.T.  and CONWAY,  R.A.   (1974).   Brine shrimp  bioassay and
seawater BOD of petrochemicals.   _J. Water Pollut. Cont.  Fed.   46(1) : 63-77.
                                      6-18

-------
ROUBAL, W.T., STRANAHAM, S.I. and MALIN, D.C.  (1978).  The accumulation of low
molecular weight aromatic hydrocarbons of crude oil by coho salmon  (Oncorhynohus
kisutch) and starry flounder (Platichthys stellatus).  Arch.  Environ. Toxicol.
7: 237-244.

SAUER,  T.C.,  JR.  et al.  (1978).   Volatile liquid hydrocarbons in the surface
waters of the Gulf of Mexico.  Mar.  Chem.  7_:  1-16.   (Cited in Syracuse Research
Corporation,  1980).

SAUNDERS, J.R.  (1977).  Degradative  plasmids.   Nature.   269: 470.

SEXTON, K. and WESTBERG, H.   (1980).  Ambient  hydrocarbon  and ozone measurements
downwind of  a large automotive painting plant.  Environ.  Sci.  Teehnol.  14: 329.
(Cited  in Syracuse  Research Corporation,  1980).

SLIMAK,  M.    (1980).    Exposure  Assessments  of  Priority  Pollutants:   Toluene.
Report  (draft) prepared  by Arthur D.  Little,  Inc., MA.    Prepared  for U.S.
Environmental   Protection  Agency,   Monitoring   and  Data  Support  Division,
Washington,  DC.

SMITH,  R.V.  and ROSAZZA, J.P.  (1974). Microbial models of mammalian metabolism.
Aromatic hydroxylation.   Arch. Biochem.  Biophys.  161: 551-558.

SRC (SYRACUSE  RESEARCH CORPORATION).   (1980).   Hazard  Assessment  Report  on
Toluene.  1st Draft.  Prepared for U.S. Environmental Protection Agency, Research
Triangle  Park,  NC.

 STORMER,  K.   (1908).   Uber  die wirkung des  schwefekkohlenstoffs  und ahnlioler
stoff auf den boden.  Abl. Bakt.  (Abst. 2).  20_: 282.   (Cited in Claus  and Walker,
 1964).
                                       6-19

-------
SUBRAMANIAN,  V,  SUGUMARAN,  M.  and  VAIDYANATHAN,  C.S.     (1978).    Double
hydroxylation   reactions   in   microorganisms.      ^J.    Indian   Inst.   Sci.
60(6): 173-178.

TABAK, H.H., QUAVE,  S.A.,  MASHNI,  C.I. and EARTH, E.F,  (1981).  Biodegradability
studies with  organic  priority pollutants compounds.  _J.  Water Pollut. Control
Fed. 53:   1503-1518.

TAUSSON,  W.O.   (1929).   Uber die oxydation  der benzolkohlenwaserstoffe  durth
bakterien.  Planta.  7_: 735.  (Cited in Claus and Walker, 1964).

U.S.  EPA   (U.S. ENVIRONMENTAL PROTECTION AGENCY).   (1979).   Fate of Priority
Pollutants in Publicly Owned Treatment Works—Pilot,  Study,  Publication No.   EPA
440/1-79-300.   Performed by  Feiler,  Burns  and  Roe  Industrial Services Corp.,
Paramus,  NJ.

U.S.  EPA  (U.S.  ENVIRONMENTAL  PROTECTION  AGENCY).   (I960).    Guidelines  and
Methodology  for the  Preparation  of Health Effect  Assessment Chapters  of  the
Ambient Water Quality Criteria Documents.   U.S.  EPA,  Environmental  Criteria  and
Assessment Office;  Office  of Health and  Environmental Assessment;  Office of
Research  and Development, Cincinnati, OH, November 28,  1980.

VETTH, G.D. et al.   (1979).  Measuring and estimating  the bioconcentration factor
of  chemicals  in fish.   _J.  Fish  Res.  Board Can.  j?6_:'91.   (Cited in Syracuse
Research  Corporation, 1980).

WAGNER, R.  (191*0.  Uber benzol  bakterien.  Z. Gar Physiol.  4_:  289.   (Cited in
Claus and Walker, 1964).

WALKER,  P.   (1976).   Air Pollution Assessment  of Toluene,  Publication  No.
MTR-7215.    Prepared  by the Mitre  Corp.,  McLean, VA,   under  Contract  No.
68-02-1495.  U.S.   Environmental  Protection Agency,  Research Triangle  park,  NC.
Available from:  National Technical  Information Service,  Springfield,  VA  (NTIS
PB  256-735).   (Cited in Syracuse  Research Corporation,  1980).
                                      6-20

-------
WEI, K.S. and ADELMAN, A.H.  (1969).  The photooxidation of toluene.   The role  of
an excitec! charge-transfer complex,  Tetra. Lett.  38:  3297.   (Cited  in  Syracuse
Research Corporation, 1980).

WILLIAMS,  P.A.  and WORSEY, M.J.   (1976).  Ubiquity of plasmids in coding for
toluene and xylene metabolism  in  soil  bacteria:   Evidence for the existence  of
new TOL plasmids.  J. Bacteriol.   125(3):  818-828.

WILSON,  J.T.,   ENFIELD,  C.G.,  DUNLAP,  W.J.,  CROSBY*,   R.L.,  FOSTER,  D.A.  and
BASKIN, L.B.   (1981).   Transport  and  fate of selected organic pollutants in a
sandy soil.  J.  Environ. Qual.   10: 501-506.

WORSEY, M.J. and WILLIAMS,  P.A.  (1975).  Metabolism  of  toluene and xylenes  by
Pseudcmonas  putida (arvilla)  mt-2.    Evidence  for a  new function  of  the  TOL
plasmid.   J_.  Baoteriol.   124(1) ;  7-13.

WORSEY,  M.J.,  FRANKLIN,  F.C.H. and WILLIAMS,  P.A.   (1978).   Regulation of the
degradative pathway enzymes coded for by the TOL  plasmid (pWWO) from  Pseudomonas
putida mt-2.  _J. Bactet-iol.   13M3) : 757-764.

YANO, K. and T. NISHI.  (1980).  pKJ1,  a naturally occurring conjugative  plasmid
coding for toluene degradation and  resistance to streptomycin and sulfonamides.
_J.  Bacterial.   113: 552-560.
                                      6-21

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               7.  ENVIRONMENTAL AND OCCUPATIONAL CONCENTRATIONS

     Monitoring data for the concentration of toluene has been divided into two
subsections, one pertaining  to the environmental  levels  and the  other  to the
occupational levels.
7.1.  ENVIRONMENTAL LEVELS
     Toluene has been detected  in  the  following environmental  media:   (1) air,
(2) aqueous media,  (3) sediments,  (4) solid wastes and leachates, and (5) edible
aquatic organisms.
7-1.1.   Air.   Toluene  is the  most  prevalent aromatic hydrocarbon  present  in
ambient air.  Atmospheric levels of toluene in different locations in the United
States and other parts of the world are given in Table 7-1.  The quantification
of  toluene  levels  in  the  atmosphere  has  exclusively  been  done by  GC-FID,
especially with capillary columns  (Section 8.1.1.3).
     From the experimental measurements  of the  toluene-to-benzene  ratio,  Pilar
and Graydon  (1973)  concluded that  the major source of toluene in urban air with
high  traffic volume is  automobile emission.   Recently,  Pellizzari  (1979)  has
measured  toluene  levels near  manufacturing  and  refining sites in  the  United
States.  The ratio of toluene  to  benzene in  these sites indicates  that besides
automobile  emission,  manufacturing processes are probably a  factor  in ambient
toluene concentration at many of the sites.
     It can  be  inferred from  Table  7-1  that the  atmospheric  concentration  of
toluene in u~ban areas not  containing toluene manufacturing or gasoline refining
sites are in the same range as the sites containing these  industries.  It can be
concluded  also  from Ta-bl-e- 7-1  that  the  concentration of  toluene  has declined
significantly in the past  15 years  in Los Angeles, presumably as  a  result  of
automotive emission controls.  The concentration of toluene in many urban areas
in the United States in recent years ranged from less  than 0.1  ppb  to as much as
50 ppb, averaging approximately  1  to 10 pp'o.   In remote locations of the United
States, the  value  averaged approximately 0.3 ppb in  1971  (Table 7-1),  but the
current level (data reported in  1979)  may be  lower  as indicated by the toluene
concentration at Grand Canyon,
     Sexton  and  Westberg (19BO) monitored the  air  near  an automotive painting
plant at Janesville, Wisconsin, to investigate the effect  of emission from paint
solvents  on atmospheric  toluene  level.   The  toluene  concentration downwind
                                      7-1

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I
ro
                                                           TABLE  7-1


                                             Atmospheric  Concentrations  of Toluene
Concentration, ppb
Location
Urban Areas :
Azusa, CA

Azusa, CA

Baton Rouge, LA
Eatsto, NJ

Bayonne , NJ

Birmingham, AL
Burnett, TX

Camden , HJ

Deer Park, TX
Sampling
date

1967

1975

1977
1979

1969

1977
1977

1979

1974-1977
Median or
Average

14

13

0.15
0.6

11.8

2.0
30.0

6.97

67
Highest or Range

23

5.9-31

0.05-0.19
ND-3.5

NR

0.21-1). 7
NR

0.23-38

3.2-99
Reference

Altshuller et al.,
1971
Mayfsohn et al.,
1976
Pellizzari, 1979a
Bozrzelli et al.,
1980
Lonneman et al.,
1974
Pellizzari, 1979a
Oldham et al. ,
1979
Bozzelli et al.,
1980
Oldham et al.,
                                                                                                     1979

                                                                                                     Lonneman  et  al.,

                                                                                                     1979

-------
TABLE 7-1  (cont.)
Concentration, ppb
Location
Denver , CO
Denver City, TX
Edison, NJ
El Dorado, AR
Elizabeth, NJ
El Monte, CA
El Paso, TX
Houston, TX
Irving, TX
Jacinto City, TX
Sampling
date
1973
1973-1980
1977
1976
1978
1978-1979
1975
1978
1973-1980
1977
1973-1974
Median or
Average
9
8.1
1000
350
9.7
7.5
16.0
5.7
10
9.5
18
Highest or Range
74
0.71-37
70-5500
NR
0.12-39
ND-85
2.9-51
1.0-99
0.21-53
NR
6.3-29
Reference
Russell, 1977
Singh et al.,
1980
Russell, 1977
Oldham et al. ,
1979
Pellizzari, 1977
Psllizzari, 1979a
Pellizzari, 1979a;
Bozzelli et al.r
19PO
Mayrsohn et al.,
1976
Pellizzari, 1979a
Brodzinsky and
Singh, 1982
Oldham et al.,
1979
Lonneman et al.,
                                             1979

-------
TABLE 7-1 (cont .)
Concentration, ppb
Location
Jones State Forest, TX
La Porte, TX

Lake Charles, LA
Liberty Mound, OK

Linden, NJ

Long Beach, CA

Los Angeles, CA













Sampling
date
1978
1973

1978
1977

1969

1975

1963-1965

1966

1967

1968

1971

1973

1979

Median or
Average
1. t
5.6

0.33
0.98

15.0

6.7

59

37

30

39

50

22

1 i.7

Highest or Range
0.6-13.1
m

0.08-0.58
0.07-9.9

m

1.1-23

m

129

50

NR

m

m

1.1-53.1

Reference
Seila, 1979
Oldham et al . ,
1979
Pellizzari, 1979b
Eaton et al.,
1979
Lonneman et al.,
1971
Mayraohn et al.,
1976
Leonard et al.,
1976
Lonneman et 11 .,
1968
Altshuller et al.,
1971
Kopeznski
et al., 1972
Altshuller et al.,
1971
Leonard et al .,
1976
Singh et al . ,
1979

-------
TABLE 7-1 (cont.)
Location
Magtaa, UT
Manhattan, NY
Newark, NJ
Newbury Park, CA
Oakland, CA
Pasedena, TX
Philadelphia, PA
Phoenix, AZ
Rio Blanco County, CO
Riverside, CA
Sampling
date
1977
1969
1979
1978
1979
1973-1974
1979
1979
1978
1970-1971
Concentration, ppb
Median or
Average Highest or Range
0.33 0.23-0.43
13-5 NR
2.6 0.01-13
NR 0.7-13
3.1 0.15-16.9
25 2.4-46
4.5 2.1-5.7
8.6 0.54-38.7
1.2 0.7-2.5
NR 9-18
Reference
Pellizzari, 1979a
Lonneman et al . ,
1974
Bozzelli et al. ,
1980
Hester and
Meyer, 1979
Singh et al.,
1979
Lonneman et al^
1979
Westberg and
Svreany, 1980
Singh et al. ,
1979
Arnts and
Meeks, 1981
Stephens, 1973

-------
TABLE 7-1  (cont.)
Location
Riverside, CA

Rutherford, NJ

S. Charleston, VW
South Amboy, NJ

Sperry, OR

St. Louis, MO

Troy, NY

Tulsa, OK

Tuscaloosa, AL

Upland, CA

Vera, OK
Sampling
date
1980

1979

1977
1979

1977

1980

NR

1977

1977

1975-1977

1977
Concentration, ppb
Median or
Average Highest or Range
5. a 3.0-9.0

8.14 0.01-33

0.05 0. OH -0.07
2.2 ND-9.7

1.4 0.145-4.8

1.5 0.2-2.6

1.0 NR

1.6 0.04-13

38 24-85

9.9 0.8-38

0.81 0.26-1.8
Reference
Singh et al.,
1980
Bozzelli et al.,
1980
Pellizzari, 1979a
Bozzelli et al.,
1980
Eaton et al.,
1979
Singh et al.,
1980
Atwicker et al.,
1977
Brodzinsky and
Singh, 1982
Holzer et al.,
1977
Brodzinsky and
Singh, 1982
Eaton et al., 1979

-------
                                                        TABLE 7-1  (cont.)
-i
 i
Location
Wyona, OK
Rural and Remote Areas:
Brethway-Gunderson Kill, WA
Camel's Hump, VT
Hell's Canyon, ID
Moscow Mt., ID
Point Reyes, CA
Grand Canyon , AZ
Talladega National Forest, AL
Sampling
date
1977

1971
1971
1971
1971
1971
1977
1977

Median or
Average
0.30

0.01
1.0
0.3
0.2
0.2
Trace
0.14
Concentration, ppb
Highest or Range
0.09-0.7

NR
NR
NR
NR
NR
Trace
0.2-1.3
Reference
Eaton -et al.

Robinson et
1973
Robinson -et
1973
Robinson et
1973
Robinson et
1973
Robinson et
1973
Pellizzari,
Holzer et al

, 1979

al.,
al.,
al.,
al.,
al.,
1979a
• i
       Srnokey  Mountain,  TN
1978
0.96
0.3-2.4
1977


Arnts and Heeks,

1981

-------
                                                TABLE 7-1  (cont.)
Location
Global:
Zurich, Switzerland
Toronto, Canada
Berlin, W. Germany
i -Stockholm, Sweden
The Hague, Nether land
Helsinki, Finland
Gatwick Airport, England
Sampling
date

NR
1971
1975-1976
NR
1974
1979
1979

Median or
Average

39
30
27
NR
18
NR
58.6
Concentration, ppb
Highest or Range

NR
188
2. if -94. 2
0-2.7
5U
15.9-37.1
1.2-809.6
Reference

Grob and Grob,
1971
Pilar and Graydon,
1973
Lahmann^et al.t
1977
Johansson, 1°78
Leonard et al.,
1976
Hasanen et al . ,
1981
Tsanl-Bazaca
et al., 1982
ND = Not Detected
NR = Not Reported

-------
within 1.6 km of-the plant was 160 ppb.  The concentration of toluene was still
20.5,  22.9,   17.5,  and  15.1 ppb  at  distances  6,  10.5,  13.5,   and   16.5 km,
respectively, downwind from  the  plant.   These concentrations are  about  10 to
15 times higher than the  background toluene concentrations of  U5  ppb determined
at a  distance of 1.6 km upwind  of  the  plant.  These  concentrations  are also
comparable to or higher than most  of the values given in  Table 7-1.
     In response to  numerous  complaints from residents about illness and  odors in
the vicinity of a chemical solvent reclamation plant in Maryland,  Smoyer et al.
(1971) monitored the valley  air surrounding the plant.  A toluene  concentration
as high as 11 ppm was registered in the valley r.ir.  Both this result and  the more
recent  investigation  of  Sexton  and  Westberg  (1980)  indicate  that  processes
involving solvent use of  toluene  may result in high emission of  toluene In the
vicinity of  these sources.
7.1.2.  Aqueous Media.  Toluene has  been monitored in a number of aquatic media
including:    (1) surface  waters,   (2)  industrial wastewater,   (3) water  from
publicly-owned  treatment   works  (POTW),   (*4) underground  waters,   (5) municipal
drinking waters, and (6)  rainwater.  The toluene levels in each of  the media have
been discussed separately.
     7.1.2.1.  SURFACE WATERS — Information regarding toluene levels in surface
water  has been obtained primarily  from the STORET system as reported by Little
 (1981).  Table 7-2  shows the toluene levels for major river basins in the United
States.  It is evident from Table 7-2 that 33?  of all  the monitored surface water
contains  toluene  levels  below  a  concentration of 10 ppb.  The concentration of
toluene  in  surface  water?  of the  central  region  (Lake Erie,  upper Mississipi,
Lake  Michigan,  etc.)  are  higher than  surface  waters from other  regions.  This
higher level of toluene cannot  be attributed  to  the  emission from production
sites  since  the  central region contains only 8 of the 38 major production sites.
 Surface waters from Texas,  which  contain 20 of the 38  production  sites, showed
 lower  levels of toluene.  This indicates that  production processes may not be the
major  source of  toluene emission in  surface waters.
      Recent  studies of the  coastal waters  of  the  Gulf  of  Mexico  have shown  that
aromatic hydrocarbons comprise 80  to 90?  of the total  dissolved  volatile hydro-
carbons  (
-------
                                    TABLE 7-2

Distribution of U.S; Surface Waters Within a Certain Toluene  Concentration  Range

Northeast
North Atlantic
Southeast
Tennessee River
Ohio River
Lake Erie
Upper Mississippi
Lake Michigan
Missouri River
Lower Mississippi
Colorado River
Western Gulf
Pacific Northwest
California
Great Basin
Puerto Rico
Utjlabeled
TOTAL
Number of
Observations
1
14
110
16
54
2
18
30
34
8
3
15
80
5
1
1
1
393
Percent Sample in the Toluene Concentration
Range, ppb
<10 10.1-100 100.1-1000 >1000
100
100
93 4 4
81 6 66
98 2
100
67 22 11
20 77 3
44 53 3
88 13
100
100
99 1
100
100
100
100
83 14 3 IA
 Source:   U.S.  EPA,  1980a
1A   insignificant  amount.
                                        7-10

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     7.1.2.2.  INDUSTRIAL WASTEWATERS — Table  7-3 shows the levels of toluene in
industrial effluents as stored in the  STORET  system (Little^  1981).   It can be
concluded from Table 7-3 that 85% of the effluents showed toluene concentrations
of less than 10 ppb.  Fifteen  of the reporting stations showed toluene concen-
tration in excess of 100 ppb.
     Wastewaters from a speciality  chemicals manufacturing plant were analyzed
by Jungclaus et al. (1978).  The concentration of toluene in the wastewater was
reported to be in the  range of  13 to  20 ppm.  Similarly, wastewater from one tire
manufacturing company was  analyzed  by  Jungclaus  et  al. (1976)  and was found to
contain approximately  10 ppm of  toluene.   Both of  these  values  are among the
highest values reported in Table 7-3.
     An analysis of raw wastewater  and secondary effluent from a -textile manu-
facturing  plant  was  reported  to  contain toluene  as  one  of  the  predominant
compounds  (Rawlings  and  Samfield,   1979).   The  toluene  concentrations  in  22
wastewater samples and 22 secondary treated effluent samples were in the range of
0.5 to  300  ppb  (Rawlings and Samfield, 1979).   Effluents  from a paper mill in
Biro  Bay,  Japan, were  analyzed for  organic matter.   It was  determined that
toluene constituted  1?  of the total  chloroform  extractables  from the effluent
(Yamaoka and Tanimoto,  1977).
     Toluene  has also  been  detected  in  a variety  of industrial wastewaters.
Table 1-^ shows  the frequency  of  toluene detection  in industrial wastewaters
(U.S. EPA,  1980a)'.
     7.1.2.3.   PUBLICLY-OWNED TREATMENT  WORKS (POTW)  — A pilot  study  of two
POTWs, one handling more or,-anic pollutant than the  other, was conducted for the
U.S.  EPA (1979).  Toluene  was detected in  100?  of the influent  samples and  95$ of
the final  effluent samples from the  plant  containing more organic pollutants.
The maximum  and  median  toluene concentrations in the  influent sample from this
plant were 140 and 13 ppb, respectively.   The influent sample at the other plant
had maximum and median toluene  concentrations of  37 and  10 ppb,  respectively.
The frequency of toluene occurrence  at this  plant  was 76? for the influent and
T\% for the  final effluent  sample.
     The state of Ohio (U.S. EPA, 1977) conducted a survey of toxic substances in
2 municipal wastewater treatment plants.  The toluene concentration in  the waste-
water of  the plant dealing primarily with domestic  wastewater ranged between
1 and 5 ppb.  The treated effluent from the same plant, on  the other hand,  showed
a concentration of 1 ppb.  About 87? of the influent  from the other plant which
                                      7-11

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                                    TABLE 7-3
                 Percent Distribution of U.S.  Wastewaters Within
                     a Certain Toluene Concentration Range
Effluent
Discharged
Northeast
North Atlantic
Southeast
Tennessee River
Ohio River
i
Upper Mississippi
Lake Michigan
•Missouri River
Colorado River
Western Gulf
Pacific Northwest
TOTAL
Number of
Observations
103
48
100
28
70
64
6
16
1
1
45
482
Percent
<10
84
88
87
96
84
69
100
100
100
100
91
85
Sample in the
Range ,
10.1-100
9
6
10
4
11
30




7
11
Toluene Concentration
ppb
100.1-1000
4
6
3

3
2




2
3
>1000
3



1





1
Source:   U.S.  EPA,  1980a
                                    7-12

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Industry
                                    TABLE 7-4
            Detection Frequency of Toluene in Industrial Wastewatersc
Frequency of Detection
(No. Found/No. Samples)
Soap and Detergents
Adhesives and Sealants
Leather Tanning
Textile Products
Gum and Wood Products
Pulp and Paper
Timber
Printing and Publishing
Paint and Ink
Pesticides
Pharmaceuticals
Organics and Plastics
Rubber
Coal Mining
Ore Mining
Steam Electric Power Plants
Petroleum Refining
Iron and Steel
Foundries
Electroplating
Nonferrous Metals
Coil Coating
Photographic
Inorganic Chemical
Electrical
Auto and Other Laundries
Phosphates
Plastic Processing
Procelain Enameling
Landfill
Mechanical Products
Pubicly-Owned Treatment Works
       1/20
       2/11
       19/81
       56/121
       11/18
       4/98
       58/285
       50/109
       23/147
       38/95
       306/723
       15/67
       53/249
       6/72
       32/84
       18/76
       43/431
       2/54
       5/18
       21/173
       2/12
       9/25
       10/107
       1/35
       9/56
       1/33
       1/1
       2/19
       3/17
       23/35
       11/40
 Source:   U.S.  EPA,  1980a
                                   7-13

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treated industrial-domestic  wastewater showed  the  presence cf  toluene  in the
concentration range of 8 to  150 ppb.  The frequency of  toluene detection in the
treated effluent from  the same p.1 ant amounted to 36%.   The toluene concentrations
in these treated effluents ranged from  1 to  10  ppb.
     7.1.2.4.  UNDERGROUND WATER — The Mew  York State  Department of Health and
the United States Geological  Survey examined  39  wells  in 1978 for  organic contam-
ination in groundwater (Little,  1981).   Toluene was  detected in 85$ of the wells
tested.  However, the toluene concentration,  in  these  waters  was  below 10 ppb.
     Toluene concentration in well water can be obtained from the data recorded
in STORET (U.S.  EPA,  1980a).  Eighty seven percent  of the monitored data showed
less than 5  ppb  (detection  limit)  toluene.   Of the  T43 monitored  data,  only 3
indicated the  presence of  toluene  in the concentration range of H2 to 100 ppb.
All of chese 3 wells were in the vicinity of landfill sites.
     7.1.2.5.   DRINKING WATER  —  Toluene has been detected  in raw water and in
finished water supplies of several communities  in the United States.  Levels of
up to  li ppb were  found  in finished water from the New Orleans area (U.S.  EPA,
1975a).   In  a  nationwide  survey  of  water  supplies from  10  cities,   6  were
discovered to be contaminated with toluene (U.S. EPA,  1975b). Concentrations of
0.1 and 0.7 ppb were measured in 2  of these water supplies and were detected but
not quantified in the rest.  Toluene was detected but not  quantified in 1 of 111
finished drinking waters during  a second nationwide survey  (U.S.   EPA, 1977).  In
a  subsequent  phase  of this  survey, toluene  was  found in  1  raw water  and  3
finished waters out of 11  supplies  surveyed  (U.S.  EPA, 1977).  A  level of 19 ppb
measured by gas chromatography/mass spectrometry was found in 1 of these finished
waters, and  0.5  ppb was found in another.
     In a survey of volatile organic compounds in water at  30 Canadian potable
water treatment facilities, Otsun et  al. (1982) detected toluene  in the raw water
with a frequency of 15$ and  in  the treated water with a frequency of 20$ during
the months of  August  and  September.   The average arid maximum concentrations of
toluene  in  treated  Canadian water  were reported  to  be  2 ug/Z. and  27 ug'K,
respectively.   The  corresponding  values for  the raw  water  were  <1  (ig/S.  and 15
[ig/L, respectively.  The  frequency  of occurrence and the concentration of toluene
in water showed seasonal variation, with the summertime values found  to be higher
than the wintertime values.
     Nineteen  volatile  organic compounds, including  toluene,  were detected at
concentrations  below  5 ppb  in  District of  Columbia drinking  water  (Saunders
                                      7-11

-------
et al.,  1975).  These  investigators also found that  the  concentrations of the
various contaminants  in tap water  varied by uns-pecified  amounts  from Week to
week, but the chemic?.! composition remained the same.
     7.1.2.6.  RAINWATER — Toluene has been detected in rainwater from Berlin,
West Germany (Lahmann  et  al., 1977).  The toluene content in the rainwater varied
with  sample collection  points.   The  rainwater  from  a   residential  area,  an
airport,  and a  busy  traffic  intersection  showed  toluene  concentration  of
0.13 ppb, 0.70 ppb, and 0.25 ppb, respectively.
7.1.3-   Sediment.   Toluene concentrations  in sediment samples  as  recorded in
STORET (U.S. EPA,  1980a) show  that 91$  of  the samples contain less than  10 ppb of
toluene.   The  concentration  of  toluene  exceeded  500 ppb in  only  7»  of the
samples.  Samples  with  higher  concentrations (greater than 500 ppb) of toluene
were obtained from the vicinity of an  industrial area in San Francisco.
     Jungclaus  et  al.  (1978)  monitored  the sediment  from  a  river  receiving
industrial  effluent  from a specialty  chemicals manufacturing  plant containing
toluene.  However, these investigators  could not detect the presence of toluene
i'n the river sediment.
7.1.H.  Edible Aquatic Organisms.   Of  the 59 monitored tissue samples  that were
recorded in the STORET system (U.S. EPA,  1980a), 955  of the data showed toluene
concentrations of less than 1  ppm.  The maximum toluene concentration  detected in
1  fish  tissue was 35  ppm.   It could  not be determined  whether these concen-
trations  were  determined in  edible flesh or in whole  fish.   Toluene was also
detected in fish caught  from  polluted  waters in the proximity of petroleum and
petrochemical plants in Japan (Ogata and Miyaki, 1973).  A concentration of  5 ppo
was measured in tne muscle of 1 such fish.
7.1.5.  Solid Wastes  and Leachates.  Toluene has been  detected in the  air samples
at a few landfill  sites (U.S.   EPA,  1980b) and in well water near a few landfill
sites  (U.S.  EPA,   1980c).   However,  no data  regarding  the level of toluene in
solid wastes and their  leachates  could  be found in  the literature.
7.2.  OCCUPATIONAL CONCENTRATIONS
     Several  reports   describing  the  presence  of   toluene   in  occupational
atmospheres  were   found  in  the  literature.    A   toluene level  of  10,000 to
30,000 ppm was reported in a merchant ship after it  was internally sprayed with  a
toluene-containing insecticide  (Longley  et al.,   1967).   Two  hours  after the
initial  monitoring,  concentrations ranging  from  5000 to   10,000 ppm were  still
present in the atmosphere of the  ship.
                                      7-15

-------
      A  monitoring program was instituted in response to a report of an epidemic
solvent  poisoning in a rotogravure  plant in Milan,  Italy.   Solvent containing
toluene was  largely used  in this  plant  as an  ink  solvent and diluent.   The
results  of  the monitoring showed  that the concentration of toluene ranged from 0
to  277 pprr  in  difl'erent  parts  of  the  work areas  (Forni  et al.,  1971).   The
                                          i
determined  toluene  concentrations  at different  parts  of the plant  during the
period  1957 to  1965 are shown in Table 7-5.
                                   TABLE 7-5
                 luluciic Cuin.o-ii.ra1>. iGn;>  J.U I/ iffei ciii.  Wui iv  Ai'caS
                    of a Rotogravure Plant in Milan,  Italy3
Work Area
Center of Room
Folding Machines
Between Machine Elements
Toluene
Range
1140-239
56-277
3J6~32'4
Concentration, pptn
Annual Mean
203
203
J431
Source:   Forni et al.,  1971
      In "96C, the atovs rotogravure plant was me1""1 to a different location and
 the ventilation  system of  the  plant  was improved.   Subsequent  analysis for
 toluene showed annual  mean  concentrations at  156 an'l  ?65  ppm near the folding
 machines and between  the machine elements, respectively (Forni et al., 1971).
      Tolu«ne  exposure  levels  for  other  occupational  groups   are   shown  in
 Table 7-6.  Many  of the levels given in this  table either originate  from exposure
 evaluation in foreign countries, or the data may be  too old to have relevance in
 contemporary working  atmospheres.   Accidental exposures to  toluene  are  discussed
 in Sections  11.1.1.1.  and 11.1.1.2.1.
      A study of 8 Japanese factories  operating  polychromic  rotory  processes fcr
 photogravure  printing  reported  toluene  concentrations  in the  range of  U to
 240 ppm in different  work areas of the  plants  (Ikeda and Ohtsuji,  1969).
      Toluene  exposures to  an  unspecified   number  of workers in  11 leather-
 finishing  and rubber-coating  plants   have  also been reported  (Pagnotto and
 Lieberraan, 1967).  Toluene is used  as  a lacquer thinner and  stain remover in the
 leather finishing  industry.    In  ruboer-coating plants,  the major  source of
                                       7-16

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                                    TABLE 7-6

            Toluene Exposure Levels for Different Occupational Groups
Exposure Level
Type of Occupation
Reference
100-1100 ppma

150-1900 ppmb


3-211 ppm

30.6 ppm (mean)0

80-300 ppm

15-200 ppm (mean)

50-1500 ppm

200-400 ppm

300-430 ppm

200-100 ppm

18-500 ppm

56-824 ppm

100-200 ppm  (TWA)
  occasional rises
  to 500-700 ppm

16-164 ppm

21-187 ppm

7-112 ppm (TWA)
Airplane painting

Paint and pharmaceutical
  industry

Automobile painting

Automobile painting

V-belt manufacturing

Shoemakers

Not stated

Rotogv-avure printing

Rotogravure printing

Rotogravure printing

Rotogravure pringlng

Rotogravure printing

Rotogravure printing



Rotogravure printing

Rotogravure printing

Rotogravure printing
Greanburg et al., 1942

Parmeggiani and Sassi, 1954


Ogata et al., 1971

Hanninen et al.,  1976

Capellini and Alessio, 1971

Matsushita et al., 1975

Wilson, 1943

Banfer, 1961

Munchinger,  1963

Suhr,  1975

Szadkowski et al., 1976

Forni  et al., 1971

Funes-Craviota et al., 1977



Ovrum  et al., 1978

Vaulemans et al., 1979

Maki-Paakkanen et al., 1980
  Paint contaminated with other  volatile  components  (Table 11-9)
  Concomitant exposure  to butyl  acetate  (Section 11.2.1)
 "Concomitant exposure  to other  organic  solvents (Taolo 11-3)
  Concomitant exposure  to 20-50  ppm  (mean gasoline in a few working places
  (Section  11.1.2)
                                       7-17

-------
 toluene  emission is the  fabric-spreading  machine areas.   The  concentration of
 toluene  in work  areas  of  these industries  is sijowri in Table 7-7-

                                    TABLE 7-7
                     Toluene Concentrations  in Work Areas of
                  Leather Finishing and  Rubber Ccatins;  Plants3

                                                     Toluene Concentration, pptc
Industry
Leather Finishing
Work Areas
Finishing Area
Washing and Topping Area
Range
19-85
29-195
Average
53
112
Rubber  Coating         Spreading Machines            3^-120                73

 Source:   Pagnotto and Liebernan,  1967
       Toluene has been detected 'in other occupational atmospheres.  Tor example, a
  toluene  concentration of 0.18 ppzn  has been reported  in  a  submarine atmosphere
  (Chiantella et al.,  1966)-   The origin of  toluene  in this atmosphere  has been
  speculated to be paint  solvents  °',d diesel fuel used in the submarine.   Toluene
  has  been  detected  in  the  atmosphere  of  M15 and  M19 antitank  cines  (Jenkins
  et al.,  1973).  The origin  of toluene in this atmosphere was attributed to mine
  casings.
       A  more recent study (Fraser and Rappaport, 1976) designed to determine the
  health  effects  associated  with  the  curing of  synthetic rubber simulated the
  vulcanization process in the laboratory.  Toluene emission in the vulcanization
  area  from  this  experiment   amounted  to 1. .1 pptn.   The  actual  field su-vey  of
  different  work areas  of 10  large  tire manufacturing  plants  across the  United
  States  was conducted by Van  Ert  et  al. (1980).  The  toluene  concentrations in
  different  work areas measured by these investigators is shown In Table 7-6.
       It  can be concluded from Table 7-8 that the extrusion process  area and the
  tire  building process area are  the  two areas  of tire manufacturing plants that
  account  for the major toluene emissions from these plants.
                                        7-18

-------
                                    TABLE 7-S

  Toluene  Concentrations  in Selected Work Areas of Tire Manufacturing Plants '
Work Area
Cement Mixing
Extrusion
Tire Building
Curing Prepa-ation
Inspection and Repair
Warehouse
No. of Plants
Surveyed
8
U
2
3
3
2
Area Toluene Concentration, ppa
Mean
2.9
14.0
8.0
0.6
1.9
0.28
Range
0.2-7.7
3-3-50.0
2. 5-13- H
0.1-1.1
0.6-2.7
0.01-0.76
aSource:   Van Ert et al.,  1980
 All of the plants,  with the exception of plants where the warehouse sanples
 were taken,  were surveyed during 1973-77.   The warehouse saaples were collected
 in 1977-
                                    7-19

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7.3.  CIGARETTE SMOKE
     The concentration  of toluene in  inhaled  cigarette smoke  is  approximately
0.1 mg/cigarette  (NRC,  I960;  Dalhamn  et al.,  1968).   Jermini  et  el.  (1976)
determined the concentration  of  toluene in the sirtestrean smoke  of  cigarettes.
When 30 cigarettes were inhaled in a  30 ns  room and  the  concentration of toluene
in room air was determined, it was found to be, 0.23  ppm.  This value  corresponds
to 0.87 ffig of toluene in the  sidestream  smoke  of  each cigarette.   Holzer et  al.
(1976) determined the toluene concentration in a 60  m  room and found an ambient
toluene concentration of <<0 ppb.   When 1 cigarette  was  smoked  in  the  room,  the
concentration of toluene rose  to  *i5  ppb.   This corresponds to 1.1  mg of toluene
contribution from each cigarette.  It  seeira from  this discussion  that  the main-
stream  smoke  of  1  cigarette  contributes  0. 10 Eg toluene to the smoker.   The
sidestreaa saoke, on the other hand, may contain  a  higher amount  of  toluene.

7.M.  REFERENCES

ALTSHULLt-R,  A. P. ,  LGNNEMAN,  W.A.,   SUTTERFIELD,  F.D.  and  KOPCZYNSKI, S. L.
''971).    Hydrocarbon  cotsposition  of  the   aUnosphere  of  the Los  Angeles
Basin—1967.   Environ-. Sci.  Tech.  5:  1009.

ARSTS,  R.R. and  HEEKr,  3.A.   (19&1).  Biogenic hydrocarbon  contribution to  the
ambient air o:' selected areas.   Atmos.  Environ.   15:  16^3-1651-

ATWICKES,  E.R.,  WHITBY,  R.A.  and  STASIUK,  W.N.  (1Q77).    Ambient  hydrocarbon
levels at two elevated and some street  level  sites.  Proc. Int. Clean  Air Congr.,
«th.  Taken fron:  Cheo. Abst.   89:HJ1039q, 1978.

BAfiFES, H.  M9ti).  [Studies  on  the effect of pure  toluene on the blood picture
of   photogravure   printers  and   helper  workers.]     Zentralbl.   Arbietsmed.
11:  3^-40.  (In Ger.)   (Cited  in  NIOSH,  1973).

B07.ZELI, J.W., KEBHEKUS, B.B.  and GREEfffiURG,  A.  (1980).  Analysis  of Selected
Toxic and  Carcinogenic  Substances in  Ambient  Air in Neu Je-sey.  State of  New
Jersey Department of Environmental Protection,  New  Jersey.
                                      7-20

-------
BRODZINSKY,  R.  and SINGH,  H.B.    (1932),   Volatile  Organic Chemicals  ir. the
Atmosphere:.   An Assessment of Available Data.  Final report.  Prepared for U.S.
EPA on  Contract  No.  68-02-3^52,   Environmental  Sciences  Research Laboratory,
Office of Research and Development, U.S. EPA, Research Triangle  Park, NC.

CAPEU.INI, A.  and ALESSIO, L.  (1971).  fThe urinary excretion of  hippuric acid
in workers exposed to toluene.]  Hed. Lavoro.  fr?:  196-201.  'In Itai.)

CHIANTELLA,  A.J.,  SMITH,  W.D., UMSTEAD, M.E. and JOHNSON,  J.E.  '1966).  Aromatic
hydrocarbons in. nuclear submarine  atmosphere.  Amer. Ind.  Hyg. _J.  March-April,
p. 186-192.

DALKAMN, T., EDFORS, M.L. and RYLANDER,  R.   (1968).  Mouth absorption of various
compounds in cigarette smoke.  Arch. Environ. Health.  16(6): 831-835.

EATON, W.C.  et  al.   (1979).  Study of the Nature of Ozone, Oxides  of  Nitrogen, and
Non-methane Hydrocarbons  in Tulsa, Oklahoma.  Vol.  II and  III.   U.S. EPA Report
No. ^50-^-75-008.  Research Triangle Institute,  Research  Triangle Park,  NC.  'As
Cited in Brodzinsky and Singh, 1982).

FORNI, A., PACIFICO, E. and LIMONTA, A.  (1971).  Chromosome studies In workers
exposed to benzene or toluene or both.  Arch-. Environ. Health.   22'. 3) : 373-378.

FRASER,  D.A.  and  RAPPAPORT,  S.    (1976).   Health aspects  of the  curing  of
synthetic rubbers.  Environ. Health  Perspect.  r/:  45-53.

FUNES-CRAVIOTA, F, et al.  (1977).   Chromosome aberrations and sister-chromatid
exchange in workers  in  chemical  laboratories and a rotprinting factory and  in
children of women laboratory workers.   Lancet.   2_:  322.

GREENBUPG, L.,  MAYERS, M.R.,  HEIMANN, H. and MOSKOWITZ,  S.   (1942).  The  effects
of exposure to toluene in industry.  J_. Amer. Med.  Ag.soc.   116:  573-578.

GROB, K.  and GROB,  G.   (1971).   Gas-liquid chromatographic/ma.ss  spectrometric
investigation of C,-C?0 organic compounds in an urban atmosphere. ^J. Chroaatogr.
62: 1-13.  (Cited in Syracuse Research  Corporation, 1980).
                                      7-21

-------
HANNINEN,  K., ESKELINEN,  L.,  HUSMAN,  K. and  NURMINEN,  M.  (1976).   Behavioral
effects of long-tens exposure to a mixture of organic solvents.  Soand. J.  Work
Environ. Health.  2(M): 24C-255.
    \
HASANEN, E. ,  KARLSSON,  V. , LEPPAMAKI,  E. and JUHALA,  M.   1981.  Benzene, toluene,
and  xylene concentrations in car  exhausts and  in  city  air.   Ataos.  Environ .
HESTER, N.E. and MEYER, R.A.  (1979).   A  sensitive  technique for measurement of
benzene and alky 1 benzenes in air.  Environ.  Sci.  Technol.   1 3( 1 ) ; 107-109.
HOLZER,, C..  SHANFIELD,  H. .  ZLATKIS,  A.,  BERTSCH,  W., JUAREN,  P.,  MAYFIELD,  K.
an i LIE3ICH, H.M.   (1977).   Collection and analysis  of  trace organic emissions
from natural sources.  ^J. Chroaatogr-   ^2 : 755-764.

IKEDA, M. and OHTSUJI,  H.  (1969).   Significance of urinary hippuric acid deter-
mination as an index of  toluene exposure.   Brit.  J.  Ind.  Hed.  26f_^).: 2ii«-2i46.

JENKINS, T.F., 0'P£ILLY, W.F.,  MUP.RrlANN,  R.P., LEGGETT,  D.C.  and  COLLINS,  C.I.
(1973).    Analysis  of  Vapors  Emitted   from  Military  Mines.     Report   No.
CRREL-Sh-193, Cold Regions Researcn and Engineering Lab,  Hanover,  NH, September,
1973-

JEKMINI, C., WEBEh, A.  and GRANDJEAN, E.  (1976).  Quantitative determination of
various gas-phase components  of the side-stream smoke of cigarettes in the room
air  as a contribution  to  the problem  of passive  smoking.    Int.  Arch.  Occup.
Environ. Health.  3_6: 169-181.

JOHANSSON,  I.   (1978).   Determination  of organic compounds  in indoor air with
potential reference to air quality.   Atmos. Environ.   12: 1371-1377.

JUNGCLAUS, G.A., GAMES,  L.M.  and  HITES, R.A.    (1976).   Identification of trace
organic  compounds  in   tire  manufacturing  plant  wastewaters.    Anal.  Chertu
»8{1?ji! 1894-1896.
                                      7-22

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JUNGCLAUS,  G.A.,  LOPEZ-AVILA, V. and HITES, R.A.   (1976).  Organic  compounds  in
an industrial wastewater:  A case study of their environmental impact.   Environ.
Sci.  Technol.  1_2_(_1): 88-96.

KOPEZNSKI,  S.L.  et  al.  (1972).   Photochemistry of  atmospheric  samples in Los
Angeles.   Environ.  Scj^  Technol.    6_: 3^2.    (Cited  in   Syracuse   Research
Corporation,  1980).

LAHMANN, E.,  SEIFERT, B. and ULLRICH,  D.  (1977).   The  Pollution  of Ambient Air
and Rain Water By Organic Components of Motor  Vehicle Exhaust-Gases,  Proc.   Int.
Clear Air Congr., kth,  p. 595-597.

LEONARD, M.J. et al.   (1976).   Effects of the motor vehicle  control program  on
hydrocarbons in the  central Los Angeles atmosphere.  jJ. Air Pollut. Cont.  Assoc.
26: 359.  (Cited in Syracuse Research  Corporation,  1980).

LITTLE,  A.D.   (1981).    Exposure Assessment   of  priority  pollutants:   Toluene.
Draft  report  prepared' by Arthur D. Little,  Inc.,  Cambridge, MA,  for  the  U.S.
Environmental Protection Agency,  Research Triangle Park,  NC.

LONGLEY, E.O., JONES, A.T.,  WELCH,  R. and LOMAEV,  0.  (1967).   Two acute toluene
episodes in merchant ships.   Arch.  Environ. Health.   JjJ_:  481-^87.

LONNEMAN, W.A., BELLARnT.A.  and  ALTSHULLER, A.P.   (1968).  Aromatic hydrocarbons
in  the  atmosphere  of   the  Los  Angeles   Basin.     Environ.   Sci.   Technol.
2( 11 _)_:  1017-1020.

LONNEMAN, W.A.,  KIPCZYNSKI,  S.L., DARLEY, P.E.  and SUTTERFIELD,  F.D.   (1974).
Hydrocarbon  composition  of  urban  air  pollution.    Environ.   Sci.   Teohnol.
8: 229-236.

LONNEMAN, W.A., NAMIE,  G.R.  and  BUFALINI, J.J.  (1979).  Hydrocarbons in Houston
Air.   Environmental Sciences Research  Laboratory, U.S. EPA,   Research  Triangle
Park, NC.  (As Cited in  Brodzinsky  and  Singh,  1982).
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MAKI-PAAKKANEN,   J.  et al.   (1980).   Toluene  exposed workers  and chromosome
aberrations.  Jour. Toxicol. Environ. Health.   (>: 775.

MATSUSHITA,  T.  et  al.    (1975).   Hematological  and  neuro-muscular response  of
workers exposed to low concentration of toluene vapor.  Ind.  Health.   13:  115.

MAYHSOHN,  H.,  KURAMOTO, M.,  CRABTREE, J.H., SOLTERM, R.D. and MANO,  S.H.  (1976).
Atmospheric  Hydrocarbon   Concentrations,   June   -  September,  1975.    State  of
California Air Resources  Board,  January 1976.  (As Cited in Brodzinsky and Singh,
1982).
        I
        i
MUNCHINGER,  R.  (1963).   Der nachweis central nervoser storungen  bei losungsmitt
elexponierten   Arbeitern.       Excerpta   Medioa  Series,   Madrid.      16-21.
2< 62} : 687-689.

NRC  (NATIONAL  RESEARCH   COUNCIL).   (1980).   The Alkyl Benzenes.  Committee  on
Alkyl Benzene Derivatives,  Board on Toxicology and Environmental  Healtn Hazards.
Assembly of Life Sciences, National Research Council.  Washington,  DC:  National
Academy Press.

OGATA,  M.,  TAKATSUKA,  Y.,  TOMOKUNI,  K.  and  MUROI,  K.   (1971).  Excretion  of
hippuric acid and m- or p-nethylhippuric  acid in the  urine of persons exposed  to
vapors  of  toluene  and  m- or p-xylene  in an exposure  chamber and  in workshops,
with   specific   reference  to   repeated   exposures.     Brit.   _J.  Ind.   Med.
28CQ: 382-385.

OGATA,  M. and MIYAKI,  Y.   (1973).   Identification  of substances in  petroleum
causing objectionable odor  in fish.  Water Res.   ]_:  1193-1501.

OLDHAM, R.G.,  SPRAGGINS, R.L,,   PARR, J.L.  and  LEE,  K.W.    (1979).  Analysis  of
Organics  in Ambient  Air.   Radian Corporation,  Au:: 'in,  TX.     (As  Cited  in
Brodzinsky and Singh, 1982).

OTSUN, R., WILLIAMS, D.T. and BOTHWELL, P.O.   (1982).  Volatile organic compounds
in water at thirty Canadian potable water  treatment  facilities*  ^J. Assoc. Off.
Anal.  Chera.  '65: 1370-1374.
                                      7-21

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OVRUM,  P.,  HULTENGREN, M.,  and LINDQUlST, T.  (1978).  Exposure to toluene  in  a
photogravure printing  plant.   Concentration  in ambient  air  and  uptake in the
body.   Scand.  ^J. Work, Environ.  Health.  4(3): 237-245.

PAGNOTTO, L.D. and LIEBE.RMAN, L.M.  (1967).  Urinary hippuric acid excretion as
an index of toluene exposure.  Amer. Ind. Hyg.  Assoc. ^J.  28: 129-134.

PARMEGGIANI, L. and SASSI,  C.  (1954).   [Occupational risk of toluene:   Environ-
mental  studies  and  clinical  investigations  of  chronic  intoxication.]    Med.
Lavoro.  45: 574-583.  (In Ital.).

PELLIZZARI, E.D.   (1977).    Analysis  of Organic  Air Pollutants  by Gas Chroma-
tography and  Mass Speotroscopy.   U.S.  EPA Report  No.  600/2-77-100,  Office of
Research and Development,  U.S.  EPA, Research Triangle  Park,  NC.   (As  Cited in
Brodzinsky and Singh, 1982).

PELLIZZARI, E.D.    (1979a).    Information on  the  Characterization  of Ambient
Organic Vapors  in  Areas  of High Chemical  Pollution.   Contract No. 68-02-2721,
Health Effects Research lab, Office of  Research and  Development, U.S.   Environ-
mental Protection  Agency,  Research  Triangle  Park,  NC.   (As cited in Brodzinsky
and Singh,  1982).

PELLIZZARI, E.D.    (1979b).   Organic Screening in Lake  Charles,  LA  Using Gas
Chromatography   Mass   Spectrometry  Computer   Techniques.     EPA    Contract
No. 68-02-2714.   Research  Triangle  Institute,  Research Triangle Park,  NC.  (As
Cited in Brodzinsky and Singh, 1982).

PILAR, S. and GRAYDON, W.F.  (1973). Benzene and toluene distribution  in Toronto
atmosphere.  Environ. Sci. Technol.  7(7): 628-631.

RAWLINGS,  G.C.  and SAMFIELD, M.    (1979).   Textile plant wastewater  toxicity.
Environ.   Sci. Technol.  13(2): 160-164.

ROBINSON,  E.   et al.    (1973).     Nonurban,  nonmethane  low  molecular weight
hydrocarbon concentrations related to air mass identification.  J.  Geophys.  Res.
78: 5345.   (Cited  in Syracuse Research  Corporation,  1980).
                                      7-25

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RUSSEL,  P.A.  (Ed)    (19'77).    penver Air  Pollution Study—1973.    Proc.  of  a
Symposium, Vol.  I  and II.   Final Report, January  197^-June 1976.   Report  No.
EPA/600/9-77/001.   Atmospheric  Chemistry  and  Physics  Div.,  Denver1  Research
Institute,  University  of Denver,   CC.    Available  through  NTIS,   Order  No.
PB-264216, Springfield, VA.

SAUER, T.C.,  JR. et  al.   (1978).   Volatile liquid hydrocarbons in  the  surface
waters of the Gulf' of Mexico.   Mar.  Chera.  7_:  1-16.   (Cited  in Syracuse  Research
Corporation,  1980).

SAUNDERS,     R.A.,    BLACHLY,    C.H.,   KORACINA,     R.A.,    LAMONTAGNE,    R.A.,
SWINNERTON,   J.W. and SAALFELD, F.E.  (1975).  Identification of volatile organic
contaminants  in Washington, DC municipal water-  Water.  Res.   99:  11^3-11^5.

SEILA, R.L.    (1979).  Non-urban Hydrocarbon Concentrations  in Ambient Air  North
of  Huston,  Texas.   U.S.  EPA Report No.  600/3-79-010.   Environmental  Sciences
Research Laboratory, Research Triangle  Park,  NC.

SEXTON,  K. and WESTBERG, H.  (1980).  Amibent  hydrocarbon and ozone measurements
downwind of a large automotive painting plant.  Environ.  Sci.  Technol.   m: 329.

SINGH,  H. B., SALAS,  L.J.,  SMITH,  A.  and  SHIGEISH,  H.    (1979).    Atmospheric
Measurements  of  Selected  TOXDC Organic Chemicals.    Interim Report prepared  for
U.S.  EPA,  Environmental  Sciences Research Laboratory,  Research Triangle  Park,
NC.   Prepared by Stanford Research  Institute, Menlo Park,  CA.

SINGH,  H.B.,  SALAS,  L.J., STILES,  R.  and  SHIGFISHI,  H.  (1980).   Atmospheric
Measurements  of  Selected  Hazardous  Organic Chemicals.    Interim Report  on  Grant
805990,  SRI  International, Menlo  Park,  CA.    (As Cited in  3rodzinsky and Singh,
1982).

SMOYER,  J.C., SHAFFER,  D.C.  and  DEWITT, I. L.    (1971).   A  program  to sample  and
analyze air  pollution  in  the  vicinity of a chemical reclamation  plant.   Inst.
Environ. Sci.  Tech. Meet., Proc.   17:  339-3^5.
                                      7-26

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SRC (SYRACUSE  RESEARCH  CORPORATION).    (1980).    Hazard Assessment  Report pn
Toluene.  1st Draft.   Prepared for  U.S.  Environmental  Protection Ageny,  Research
Triangle Park, NC.

STEPHENS,   E, R.     (1973).     Hydrocarbons  in  Polluted  Air:   Summary Report
Coordinating Research Council Report CRC-APRAC-CAPA-5-63-1,  NTIS No.  Pb-230993.
Statewide Air  Pollution Research  Center,  Univ. of  California,  Riverside, CA.
(Cited in Syracuse Research Corporation, 1980).

SUHR,  E.  (1975).  Comparative Investigation of the  State of Health  of Gravure
Printers Exposed  to  Toluene.   Gesellschaft  zur  Forderung des Tiefdrucks  E.V..,
Weisbaden, Federal Republic of Germany.  92  pp.

SZADKOWSKI,  D. et al.  (1976).  Evaluation of occupational exposure to  toluene.
Medizinische Honatsschrift.   30(1) :
TSANI-BAZACA, E.,  MCINTYRE,  A.E.,  LESTER,  J. N.  and PERRY, R.   (1982).  Ambient
concentrations and correlations of hydrocarbons and halocarbons in the vicinity
of an airport.  Chemosphere.   11 : 11-23.

U.S.  EPA  (U.S.  ENVIRONMENTAL  PROTECTION  AGENCY).  (1975a).   New Orleans Area
Water Supply  Study.   Analysis of Carbon  and  Resin Extracts,   Prepared  by the
Analytical  Branch,  Southeast  Environ. Hes.  Lab., Athens,  -GA,  for  the  lower
Mississippi River Branch, Surveillance and Analysis Division,  Region  VI.   (Cited
in Syracuse Research Corporation, 1980).

U.S.  EPA  (U.S.  ENVIRONMENTAL  PROTECTION  AGENCY).     (1975b).     Preliminary
Assessment  of Suspended  Carcinogens  in Drinking Water.   Report to Congress,
Washington, DC.  (Cited  in Syracuse Research  Corporation,  i960).
                                      7-27

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U.S. EPA  (U.S.  ENVIRONMENTAL  PROTECTION AGENCY).   (1977).   National Organic
Monitoring Survey,  General  Review of  Results  and Methodology:   Phases I-III.
(Cited in Syracuse Research Corporation, 1980).

U.S. EPA  (U.S.  ENVIRONMENTAL  PROTECTION AGENCY).   (1979).   Fate  of Priority
Pollutants in Publicly Owned Treatment Works—Pilot Study,  Publication No.  EPA
HV1-79-300.    Performed by  Feiler,  Burns  and  Hoe Industrial  Services Corp.,
Paramus,  NJ.

U.S. EPA (U.S. ENVIRONMENTAL PROTECTION AGENCY).   (1980a).   STORET Water Quality
Information System, October. 1980.

U.S. EPA (U.S.  ENVIRONMENTAL  PROTECTION AGENCY).   (1980b).  Priority Pollutant
Frequency Listing Tabulations and Descriptive Statistics.   Memo from D. Neptune,
Analytical Programs to  R.B.  Schaffer,  Director  of Effluent  Guidelines Div.,
January,  1980.  (Cited in Slimak, 1980).

U.S.  EPA  (U.S.  ENVIRONMENTAL  PROTECTION AGENCY).  (1980c).   Volatile Organic
Compound (VOC)  Species  Data Manual,  2nd ed.,  Publication Ho. EPA-U50/U-80-015.
Office  of  Air,  Noise,  and  Radiation,  Office   of  Air  Quality  Planning  and
Standards, Research Triangle park, NC.

VAN ERT,  M.D.,  ARP, E.W., HARRIS, R.L., SYMONS,  M.J. and WILLIAMS, T.M.  (1980).
Worker exposures to chemical agents in the  manufacture  of rubber  tires:  Solvent
vapor studies.  Amer. Ind.  Hyg.  Assoc. _J.   4J_: 212-219.

VAULEMANS, H., VAN VLEM, E., JriNSSENS,  H. and MASSCHELEIN,  R.  (1979).  Exposure
to  toluene and  urinary hippuric acid  excretion  in a  group of heliorotagravure
printing workers.   Int. Arch. Occup. Environ. Health.   ^(Z): 99-107.

WESTBERG,  H.  and  SWEANY,  P.    (1980).  Philadelphia  Oxidant  Data Enhancement
Study; Hydrocarbon Analysis.   EPA Contract  No.  68-02-3339,  Washington State
University, Pullman, WA.  (As Cited in Brodzinsky and  Singh, 1982).
                                      7-28

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WILSON,  R.H.   (19^3).  foluene poisoning.  J Amer. Med. Assoc.  123: 1106.

YAMAOKA, Y.  and  T.  TANIMOTO.   1977.   Behavior of Organic matter  in  polluted
coastal  areas.   I,  Organic  matter in  Kraft Pulp  mill  effluent in  Hiro Bay.
Nippon.  Kagaku Kaishi, (10),  155^-1559.   (Japan). Chem. Abst No, 88: 27^032.
                                     7-29

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                           8,   ANALYTICAL METHObOLOGY

     Toluene  has  been analyzed  in  a number  of  media 'including the  following:
(1) air; (2) waters,  (3)  soils and sediments, CO  crude oil and organic solvents,
(5) biological samples,  (6) some foods, and (7) cigarette smoke.   The  analytical
Methods for the determination of toluene in each  of these media are  individually
discussed below.
8.1. AIR
     In  addition  to  the  analysis  of  test  mixtures of  toluene  in  air for  the
evaluation  of methods, toluene has also  been  deterniined  in  ambient  air, occupa-
tional  air,  forensic  air,  and air containing  the  pyrolysis  products  of organic
wastes.
8.1.1.  Ambient Air.  The determination of toluene in ambient air consists  of two
distinct steps:  sampling  and analysis.
  i   8.1.1.1.  SAILING  — Toluene can be  collected from ambient air  in several
different  ways including  grab  sanpling in  aluainized   plastic  bags Oieligan
et  al.,  1965), Tedlar bags (Altshuller et a.1., 1971; Lonrieman et  al.,  1968),  and
glass containers (Schneider et al., 1978; Pilar and Graydon,  1973).   Although the
grab  sampling is  conceptually  the  simplest  approach,  this collection  method
without subsequent concentrative technique  do^s  not provide  sufficient quantity
of  toluene  for  analytical  detection  and quantification.  Since aabient samples
contain toluene in the parts per billion range, preconcentration steps are often
necessary.
     Sample collection by cryogenic procedures (Seifert and Ullrich, 1978) is an
alternative method  for  the collection of  toluene in ambient air;  however,  the
drawbacks  of this procedure  include  the inconveniences  in sampling  and  sample
regeneration.  Also,  unless the  moisture in air  is removed,  it condenses  in tne
collection  tube and  may  reduce or restrict the  air flow through the  collection
tubes.  Various drying agents, such as anhydrone,  anhydrous K-CCU,  ascarlte,  LiH,
and molecular sieve can  be used.  It has, however, been demonstrated by Isidorov
et  al.  (1977) that  it is Impossible  to  find a drying agent  that will preferen-
tially absorb the  moisture fron air without  absorbing some of the trace organics.
     Reversible sorption on  various high  surface area  materials  provides  an
excellent  method  for preconcentrative collection of toluene from  ambient  air.
Since  the  moisture  content in the air  is  normally  3  to !) orders  of magnitude
                                       8-1

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higher than the total organics (Isidofov et al.,  1977),  the  chosen  sorbets oust
show little affinity toward moisture.   Otherwise,  the retention  capacity of the
gorberits will be reached much sooner than  desired.
     A number of sorbents  such  as Tenax GG (Holzer et  al.,  1977;  Krost  et al.,
1982), various  carbonaceous materials  (Burghardt  and  Jeltes,  1975; Holzer  et
al.,  1977;  Isidorov  et  al.,   1977),   Polisorbimid  (Isidorov  et  al.,   1977),
molecular sieves and   spherisil (Ball,  1976)',  and Porapak Q (Johansson,  1973)
have  been  successfully used.    Typically,  sampling is  performed by  drawing air
through a trdp  containing  the selected sorbent with battery-operated  diaphragm
pumps.  The air flow through the trap is controlled by needle valves and measured
by a previously calibrated  rotometer.  The trap is kept at ambient temperature to
avoid  condensation  of water.   At the  end of  the sampling,  the  trap-ends  are
closed with caps and  transferred  to  the laboratory in  a refr'.geratcd  state,  to
avoid sample loss.
      8.1.1.2.   ANALYSIS. —  The  method  of analysis  usually depends  on the method
of  sample collection.  The  earlier investigators who  used plastic  bags or glass
bottles for collection of grab aanple'3  used a trapping system for concentrating a
relatively large volume (1  t;> 10  £) of sample  before analysis.   In this  method,
the  collected, sample  is  allowed to  flow   through a  cryogenic trap  containing
suitable sorbents.   At the end of trapping, the coolant  is removed  from the trap
and. the  trap is headed quickly  tr:  vaporize and transfer the trapped  compounds
into  the gas chroaiatographic (GO columns.   The columns  used '.r/ earlier investi-
gators (Lonnesati et al., )9t>8; Aitshuller et al.,  1971)  for  aromatic separations
consisted  of Ion*,  o^en-tubuiar columns eoat.ed  with m-bis( *-p le.-ioxy-phenoxy )-
benzene combined with Apiezon grease  on a  packed  dual  column with SF-9& a^ the
liquid phase 'Filar arid Graydon,  1973V.
      The more recent,  met.nods, which use sorbents  for  trapping organics,  connect
the trap  to  a  (JC syattm  via  multiple-port gas sampling valves.   The  trap  is
heated quickly  and  the desorbed organics are  passed through the  chromatographic
columns.  Because the  collected samples contain a  multitude of  o^gani-x,, capil-
lary  columns are rioroj-illy used  for the  resolution of the organics.   The Grob and
Grob  (1971) technique, i-ivlving  th« passage  of the ther-mally desorbed organics
through a small uncoated section of the  capillary column cooled cryogenically, is
used.  When the collection is  completed, this  section  of the capillary is heated
quickly and  the sample is  separated  on  the remaining portion of the analytical
column.    A  number  of  coating  materials  for   capillary  columns  including
                                       8-2

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Emulphor ON-870  (Holzer  el  aJ .,  1977),  UCON 50  HB 2000  or  5100 (Johansson,
1978),  dinonyl phthalate (Isidorcv et al.,  1977), A1?0? (Schneider et al.,  1978),
DC-550 (Louw and  Richards,  1975), OV-17 and OV-101 (Pellizzari  et  al.,  1976) have
been used.
     In one  method,  thermal desorption  of the  organics  from the aorbents was
replaced by solvent desorption  (Burghardt and  Jeltes,  1975).   In  this  procedure,
the organics aorbed on activated carbon were desorbed by CS?.   A  part  of  the CS?
was injected into a packed  column  GC containing a long column  coated with  1,2,3-
tri-(2'-cyanoethoxy)-propane.
     The quantification  of toluene  separated  by the  GC columns  is done  almost
exclusively  by flame  ionization  detectors (FID).   Confirmation  of the authen-
ticity of the GC  peaks often is provided by coupJe'l mass  spectrometers  (MS), with
or without the aid  of a computerized  data system (Holzer  et  al., 1977;  Pellizzari
et al., 1976; Krost et al.,  1982).
     A continuous automated procedure for determining toluene  in  the ambient air
was developed by  Hester and Meyer (1979).  This method  needs no sample  preconcen-
tration prior to  analysis.   In  this method, a small diaphragm pump activated by a
timer automatically injects air into  a  1  mi gas-sampling (GS)  loop  of  a GC every
10 minutes.  The  separating column was  packed  with  Chromosorb  \f coated with N, N-
bi3(2-cyanoethyl)fonaamide.  Since  no  concentration  method  was employed, the
detector used  had  about  two orders of  magnitude higher sensitivity  than  flame
ionization detectors.   A  photo-ionization detector was found to show the required
sensitivity.
     8.1.1.3.  PR£F£RRtD METHOD  --  The  preferred  method for  the monitoring of
toluene in ambient air consists of sorbent collection,  thermal elution, and GC-
FID determination.  Collection  by  trapping toluene in a  solid  sorbent  provides a
concentration method during sample collection.   Thermal  desorption is preferred
over solvent  elution  because of  the  higher sensitivity  of the  former method.
Tenax GC is perhaps the most suitable sorbent  for sample  collection.  The collec-
tion and  thermal  deaorption efficiency of t.oluene  is  excellent  with Tenax GC.
The generation of  artifacts  during therma.1 elution i-nth Tenax GC can be  elimi-
nated largely by  proper clean-up of the sorb»".t, °r.d by  following  the GC  condi-
tioning procedure  (Kolzer et al.,  1977).  The  greatest  advantage of the ambient
sorption-thermal  elution method is its extreue  simplicity  and  speed.
     The separation and  quantification  of sorbent desorbed  components  can  be
achieved by  means  of the  GC-FID  method.  Although  photo-ionization detectors
                                      8-3

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(PID)  may have higher sensiMvity. than  flsjne ionization detectors, this higher
level  of sensitivity  is not  required  when the  samples  are precoricentrated by
solid  sorbenta.  High resolution capillary columns are a necessity because of the
observed complexity  and low  concentration of  the  samples.   Of  the different
coating  materials,   N,N-bis-(2-cyanoethyl)formamide  and   1,2,3~tris(2-cyanoe-
tncxy)-propane are probably most suitable for* the separation of aromatic compo-
nents.
     8.I.I.M.   DETECTION LIMITS — The  detection limit of toluene in ambient air
depends on the volume of air passed through the sorbent trap.  For a  25 I  sample,
the detection  limit is less than 0.. 1 ppb  (Holzer et al.,  1977) with a capillary
column and  flame ionization detector.  When direct injection (l mi) and the GC-
PID method  are  used, the  detection limit  for  toluene  is  0.3 ppb (Hester and
Meyer, 1979).
8.1.2.  Occupational Air.
     8.1.2.1.    SAMPLING —  The concentration  of toluene  in  occupational air
normally is much  higher than in ambient air.    Therefore, the  collection  of
saaples  in  certain  instances  may  not  require a  concentration  step.    The
collection  of samples by the  grab method has been used  by  a  nuraber  of authors
(Tokunaga et al.,  1971;  Chovin and Lebbe,  1967).
     Soae of the earlier methods used liquid scrubbers for absorbing toluene from
occupational air.    A  number  of  scrubbers, including potassium iodate in dilute
sulfuric acid  (Ministry  of Labour, 1966), cooled organic  solvents  such as ethyl
cellusolve  acetate,  dimethylformamide,  and dimethyl  sulfoxide  in dimethyl forma-
mide (Ogata et al.,  1975),  and nitrating  solution (Chovin and Lebbe, 1967) have
been used.   In addition  to the  inherent  limitations in  its ability to overcome
the interferences, this  method is  not convenient for  the  collection of breathing
zone samples.
     The more  recent methods used solid sorbents for the  collection of toluene.
Silicb  gel  (Ogata  et  al.,  1975;  Tokunaga et al.,  1971),   activated  carbon
(Esposito and  Jacobs, 1977; Fracchia et al.,  1977; Reid and  Halpin,  1968; Fraser
and Rapoaport, 1976;  NIOSH,  1977) and  Tenax GC  (Nimmo  and  Fishburn,  1977) are
some  of the sorbents used for this  purpose.   Aromatic hydrocarbons  such as
toluene are  easily displaced  from silica  gel  by water vapor,  resulting in pos-
sible  losses  of  toluene  in  humid  atmospheres  (NRC,  1980).    Therefore,  both
activated carbon and Tenax GC are the two most  frequently used sorbents for the
collection  of toluene from occupational  air.   The suitability of  either of the

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gorbents ia dictated by the method of sample analysis.  When thermal desorption
is used, Tenax GC is the preferred sorbent.  On the  other  hand, activated carbon
is preferred when solvent desorption is the method  used.
     8.1.2.2,  ANALYSIS — For grab samples, direct injections into a GC system
via syringes  or  gas sampling loops have  beentapplieJ  (Tokunaga  et  al.,  1974;
Chovin and Lebbe, 1967).  The separating columns used  in these cases were packed
columns with stationary  liquid  phases of  either  dioctyl  phthalate  (Tokur.^aa
et al., 197't) or bis-(beta-cyanoethyl)formamide (Chovin and Lebbe,  1967).  Flame
ionization detectors were used for the quantification of  toluene in both cases;
however,  this method  is capable  of  analyzing  toluene  in work  atmosphere at
concentrations cf around  10 ppm  (Chovin and Lnbbe,  1967).
     Toluene  collected  by scrubber methods is usually analyzed by colorimetric
methods.  Despite variations, most colorimetric methods show interferences  from
other  chemically similar  compounds (e.g.,  benzene,  xylenes, ethylbenzenes)  that
are normally  co-contaminants of  toluene.
     The  first  step in the analysis of  toluene collected in solid sorbents is
desorption.   Two methods  are  usually  available  for  desorption:   thermal  and
solvent.  Carbon disulfide is the most  frequently used solvent  for  the desorption
of toluene from solid sorbents (Esposito and Jacobs,  1977;  Fracchia  et al., 1977;
Reid and Haipin, 1968;  NIOSH,  1977; Var. Ert et al.,  I960),  although  some investi-
gators have used other solvents (Ogata  et al.,  1975).  Solvent desorption is the
method of choice when activated carbon is used as the  sorbent.  Activated carbon
has not only  high efficiency  of  reversible toluene sorption,  but  it has almost
quantitative  toluene desorplton efficiency with CS,  (Fracchia et  al.,  1977).  In
the presence of other common organic solvents found in  the  work atmosphere (e.g.,
n-butanol,  cellosolve   acetate,   butyl  cellosolve,  etc.),  the  CS,,  extraction
efficiency decreases slightly, but addition of  51 methanol  to CS2  Increases the
desorption efficiency to  almost  quantitative  value  fFracchia et al.,  1977).
     When Tenax GC  or Chromocorb  102 is  used  as  the sorbent, elution by thermal
process is the method of choice (Nimroo  and Fiahburn, 1977).  Although this method
may require multiport sampling valves  and a cryogenic  sample trap for the trans-
fer of samplts from  the sorbent trap to the GC system,  it  has certain advantages
not available to solvent elution.   Because the thermal  desorption method uses the
whole  sample  for quantification,  it has  a  higher  degree of  sensitivity than the
solvent elution method where only a small fraction  of the total sample is  used
for quantification.
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     The quantification of toluene  eluted  from solid sorbents almost always is
done by  the GC-FID method.  A number of packed GC columns have  been  used  for this
purpose.   Dioctyl  phthalate  (Tokunaga  et  al.,   197*0,  UCC W-982  (Nimrao  and
Fishburn,  1977),  N,N-bis(2-cyanoethyl)fortnamide  (Esposito and  Jacobs, 1977),
dinonyl  phthalate (Ogata et al., 1975), and Porapak Q (NIOSH, 1977) are some of
the liquid phases used for chromatographic separations.
     Other methods  of  analysis,  such as  high pressure  liquid chromatography
(HPLC) on a  reverse phase column with methanol-water as  the mobile  phase  and
ultraviolet  (UV)  detection, have been attempted (Esposito and  Jacobs,  1977), but
the sensitivity of detection was poor.
     Methods involving  the use  of detection  tubes  have  been  applied  for  the
determination  of  toluene  in  occupational  air  (Tokunaga  et  al.,  1971!).   The
accuracy of the detector tubes for  toluene  quantification is rather poor, parti-
cularly in the presence of other organic vapor  (Tokunaga  et al., 1971)).  There-
fore, the detector tubes are suitable for the rough estimation  of toluene concen-
tration in the  work  atmosphere.  More recently,  detector  tubes  designed for long-
term sampling and determination of  toluene have become  available.  A  laboratory
evaluation of a few commercially available long-tern  detector tubes was Bade by
Jentzsch and Fraser (1981).  The results indicate  that  the color development of
these tubes is isore dependent upon humidity and sampling  volume than the short-
term tubes.
     A simple directly-combined uC-Ifi  (Infrared) system was developed to detect
low molecular  weight  hydrocarbons  in air  (Louw  and  Richards,  1975).   In this
method,  the  grab sample is injected directly into a GC and the effluent  fross th
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in the case of ambient air samples, N,N-bis(2-cyanoethyl)formaraide liquid phase
will provide one of the best separations for the aromatics.
     8.1.2.4.   DETECTION  LIMIT —  The  detection  limit  for toluene  by carbon
3orpt,ion-CS^ desorption method depends on the volume of air  sampled.  Concentra-
tions as low as  0.1  ppm toluene  in  a rubber tire manufacturing factory have been
detected by this method (Van Ert et al,  1980).  For a. 100 mi eample,  the  Tenax GC
sorptlon-thennal desorption  method showed a detection limit  of 0.5 ppb (Nimmo
and Fishburn, 1977).
8-1.3-  Forensic Air.  In suspected arson cases, the method of  Twibell  and Home
(1977) can be applied to speculate  or  even confirm the cause of  fire.  According
to  this  method, nickel wires  (curie point  358°C)  coated  with finely-divided
activated carbon with  the aid of an  inert adhesive  (cement binder LQ/S6), are
suspended in the atmosphere  under test for  1  to 2 hours at room temperature.  The
apparatus is connected to  a  GC-FID  system, and the wires are heated by induction
heating.  The resulting chromatographic  profile  obtained  from  the desorbed gases
can  be compared  with  different  fire  accelerant  residues  (e.g.,  gasoline).
Although the  method is not  quantitative, it has  been  claimed to show  a betttr
sensitivity than the method  cf hot headspace analysis 'Twibell  and Hose,  1977)and
has potential for use in cases where the presence of toluene needs confirmation,
such as gasoline spills.
8.1.IJ.  Gaseous Products from Pyrolysis  of Organic  Wastes.   The  gaseous  products
from a pilot plant  burning such  organic  wastes as wood shavings, solid municipal
wastes, and  rice hull were  analy7ed  by Brodowski et al.   (1976).   The method
consisted  of collecting  grab  samples  In stainless  steel  sampling  bulbs and
injecting Q.l> m<: of  tht gaa  into a GC.  The  seoarating columns  were dual stain-
less  steel  colictns  packed  witn  Porapak QS modified  with  terephthalic acid.
Evidently, the method does not  have high sensitivity of detection.  The toluene
concentration of the  pilot  plant  gaseous products was determined  to  be 0.2 to
0.3 aol ? by this aethod rBrodowski et  al..  1976).
8.2.  WATER
     Toluene has been determined in a number of aqueous raedla including surface
waters,  industrial  wastewaters,   water  from  publicly-owned   treatment  works
(POTW), underground water, drinking water, and rainwater.
8.2.1.  Sampling.   Water  samples  other than industrial  wastewater samples are
generally collected  by  the  grab method.   In the  case of industrial discharges
where the discharge  paraaeters  are dependent on the operating  process,  continu
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ous samples using a  commercial composite sampler  have  been  used (Rawlings and
Samfield,  1979).   The preservation  and  handling  of  the  aqueous samples after
collection are especially  important for volatile  components.   The samples are
collected in glass bottles  that  are filled to overflow and sealed with teflon-
backed silicon rubber septa  and screw caps.   It has  been  demonstrated that simple
samples in non-reactive matrix (o.g.,  drinking water,  ground water) collected in
the above fashion can be held under ambient  conditions  from 10 to  22 days without
significant loss of  volatile compounds (Bellar and Lichtenberg,  1979); however,
wastewater samples  should be adjusted to  a  pH  of 2  by adding dilute hydrochloric
acid.  Any free chlorine should be neutralized by the addition of 35 mg of sodium
thiosulfate  per  1  ppm  of   free  chlorine (Federal  Register,   1979)  before  the
samples are collected in glass bottles.   The samples oust be iced  or refrigerated
during  transportation and  storage.   All  such wastewater  saaples   should  be
analyzed within 7 days of collection  (Federal  Register, 1979).
8.2.2.  Analysis.  Although  direct injection (Jungclaus et  al.,  1978) and solvent
extraction  (Jungclaus et al.,  1976)  methods  have been  used  to determine the
concentration  of orgar.ics   including  toluene  in industrial  wastewaters,  these
methods  are  not suitable  for toluene determination  in other media.   Even in
wastewater, both of these methods  have questionable accuracy.  The direct aqueous
injection method does not have good sensitivity and the solvent  extraction method
is  likely to provide low recovery  since  some  of the volatile components will be
lost  during trie concentrativo evaporating step.
      The three most  commonly used  methods for  toluene analysis in aqueous media
are  (1,1 purge and trap, (2) headspace, and (3) sorption on solid  sorbents.  Each
of  these methods is  individually  discussed below.
      6.2.2.1.  PURGE AND TRAP — Purge and trap is the most widely used method
for  the analysis of  toluene in aqueous media.   It  has been used for  the deter-
mination of toluene  in drinking waters (Bertsch et  al.,  1975; Lingg et  al., 1977;
flyan  and Fritz,  1978), in wastewatera  (Bellar and Lichtenberg,  1979; Rawlings and
Samfield,  1979; Jong:laus et al.,  1978), and  in r  inwater  (Seifert and Ullrich,
1976).   The  U.S.   Environmental  Protection  Agency recommends the  use  of this
aethod for toluene analysis in wastewater  (Federal Register,  1979).
      In this method, an inert gas (helium)  is  bubbled  through a water  sample via
a  glass  frit  contained In  a specially designed purging chamber.  The aromatics
released into the vapor  phase  are swept  through and trapped in  a sorbent tube.
After  the  purging  and trapping  is  completed,  the trap  is  transferred  to the
                                      8-8

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injection  port of a GC.  The  trap  is  heated and backflushed  into  a GC system,
where the  separation of the volatiles  takes place.   Both packed (Bellar et al.,
1979; Lingg et al., 1977; Federal  Register,  1979)  and capillary columns (Dowty
et al.,  1979; Bertsch  et  al.,  1975) employing  a variety of liquid phases have
been used.   The reso  ition  of components  can  be  expected to be  better with
capillary  columns.
     The detection  of the GC column effluents can be done either by  flame ioniza-
tion  detector  (Dowty  et  al.,  1979)   or   photo-ionization  detector  (Federal
Register,  1979). The use of photo-ionization detector will provide  better selec-
tivity and sensitivity of  detection.   The  confirmation  of GC peaks is usually
provided by mass spectrometry aided by  a computerized data system (Lingg et al.,
1977; Dowty et al., 1979; Bellar et al., 1979).
     A number of variations  of the purge-trap  method (Grob and Zurcher, 1976;
Lingg et  al.,  1977; Dowty  et al., 1979;  Bellar  et  al.,  1979)  involving  the
variation  of water  volume,  the temperature of the purging  system, the stripping
rate, the  duration  of stripping, the nature  of sorbent, and the method of desorp-
tion (thermal versus solvent) are available. Using a 5 m£ sample size and flame
ionization detection, Dowty et al.  (1979)  determined the  lower detection limit
for  toluene  to  be  0.1  ppb by  this  method.   The detection limit can be further
lowered if a  larger volume  of sample  (Lingg et  al.,  1977) or photo-ionization
detection  method is used.    The  purge-trap method is the preferred method for
monitoring toluene  in both drinking and wastewater samples.
     8.2.2.2.  HEADSPACE ANALYSIS — This method has  not  been  applied frequently
for  the analysis of field samples; however,  the method was  standardized with
water samples spiked with model compounds (Vitenberg  et al., 1977;  Drozd et al.,
1978).
     In the  method  of Drozd et al. (1978),  a  known  volume (50 mil) of water  is
introduced into a specially designed enclosed glass apparatus  (100  mJl), and the
system is thermostatically maintained  at 40°C.   After the  system  attains equi-
librium (30  minutes),  a  known volume  of headspace vapor  is  introduced into  a
capillary GC system via a trapping system consisting  of  a  short, cooled  (-70°C)
precolumn  coated with OV-101  (Grob  and Grob technique).  The separating  column  is
coated with squalene.
     The method of  headspace analysis  in the past had  faced problems owing to the
difficulty in establishing a  calibration procedure.   The  partition  coefficient
of  a component  between gas  and  liquid phases  is  dependent on the  total  ionic
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Strength.in  solution.  Therefore,  the same concentratipns of a component present
j..  two aqueous solutions  of different ionic  strengths  but otherwise identical
conditions,  will  not produce the same equilibrium vapor pressure.  This problem
of a calibration  curve  has been  largely obviated  through  the development of a
standard addition method (Drozd et al., 1978).  Water samples containing toluene
in the parts per  billion  range  can be  quantified by this method (Drozd et al.,
1^73) with a reasonable  accuracy; however,  the- method may not be applicable for
drinking water samples where the concentration'may be lower than 1 ppb.
     8.2.2.3.  SORPTION  ON SOLID SORBENTS ~ This method is rarely used for the
monitoring of toluene in  aqueous  samples.   The applicability of the method was
explored  by  Pfaender  (1976),  arid fiyan and  Fritz (1978)  used the  method for
monitoring toluene in drinking water.
     The method  consists  of passing a known  volume  of water through a sorbent
such as  XAD-2  (Pfaender,   1976)  or XAD-H  (Ryan  and  Fritz,  1978).   The sorbed
organics including toluene are desorbed either by solvent extraction (Pfaender,
1976) or by thermal desorption  (Ryan and Fritz,  1978),  and are injected onto a
GC-FID  system  for  component  separation and  quantification.   In  the  thermal
desorption method of  Ryan  and  Fritz  (1978;,  the use of a  trap  consisting of a
Tenax GC  precolumn  to eliminate the  excess  water showed a good sensitivity for
the method.   The  recovery of  toluene was  nearly  90$ when  the concentration in
drinking water ranged from 1  to  10 ppb.   For the quantification of toluene in
water by this method,  the recovery of toluene from the sorbent should be known.
8.3.  SOILS AND SEDIMENTS
8.3.1.   Sampling.   Bottom sediment samples can  be collected either by Hopper-
dredge or by clam-type dredge samplers (U.S.  EPA,  1979).  Hopper-dredge collected
samples generally contain more water  than  clam-type dredge-collected samples.
Bottom sediment samples also can  be  collected using a  core  sampler (U.S. EPA,
1979).
     For  volatile organic analysis,  the samples  should  be  collected in screw--
capped glass containers lined with aluminum foil (Jungclaus et al.,  1978)  or in
glass hypovials with  crimped aluminum  seals  and teflon-backed septa (U.S. EPA,
1979).  For best  results, the container should be filled to maximum  capacity to
reduce the amount of  headspace  and should  be  transported and stored at wet ice
temperature (U.S. EPA, 1979).
     The method of soil  sampling is given in detail by de Vera et al. (1980). The
soil samples should  be taken in a grid pattern over the  entire site.   A scoop can
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be used for collection of soil samples Up to  8  cm  deep.   To sample beyond this
depth,  a soil  auger  or Veihmeyer soil sampler, as  described by de Vera et al.
(1980), should  be used.  After the sample  is  transferred into  glass containers to
a maximum  capacity,  the  containers  must be  tightly capped with contamination-
free lids  to  prevent loss  of  volatile components  and  to exclude  possible
oxidation.    The  samples should  be  refrigerated   (4°C)  during  transport  and
storage.
8.3.2.   Analysis.  Very  few  reliable methods are available  for the analysis of
volatile organics in  soil and sediment samples.  Solvent extraction methods using
highly  volatile solvents  are  unlikely to  be  successful.    The  evaporative
concentration step of this method would result in the loss of volatile organics.
Headspace  analysis, which has  few provisions to concentrate the organics, will
produce unreasonably high detection limits.
     A modification of the purge  and trap method has been suggested by the U.S.
EPA (1979)  for  the analysis of soil and sediment  samples.   The modified purge and
trap apparatus  used  for  this purpose is  described by the U.S.  EPA (1979).  The
sample, contained  in a specially-designed  glass vial,  is  heated at  80°C and
purged with helium gas.  The desorbed organics are trapped in a  Tenax GC column.
At the end  of trapping, the Tenax GC  column is  inserted in the injection port of a
GC, and the thermally desorbed organics are analyzed by GC-FID as  in the case of
water  and  wastewater  samples.   The  recovery  of toluene  was  determined  to vary
between 32 and W% when 0,1  to  3-0 |ig of  toluene  was  spiked  onto a specially
prepared soil matrix.  Although  the  recoveries  were low, they were found to be
linear and reproducible (U.S. EPA,  1979).   Data on  spiked environmental samples
showed much higher recoveries (80 to 100?).
     With  the   purge-trap  system described,  the   minimum  detection limit  of
0.1 ppb can be  attained.   Thus, the method  showed at least two  orders of magni-
tude higher sensitivity than headspace analysis (U.S. EPA, 1979).
8.4.   CRUDE OIL AND ORGANIC SOLVENTS
     Benzene and toluene concentration in petroleum crude and other fossil fuel
samples can be  determined by  a method  developed by Grizzle and Thomson (1982).
In this method, the  acidic  and basic constituents were  removed by ion-exchange
chromatography  prior  to  fractionation  into  groups.   Alumina, chemically bonded
silica-R (NH2)2, and 2,4-dinitroanilinopropyl-silica (DNAP-silica) were  used for
liquid chromatography class  separation of aromatic hydrocarbons.  On the  basis of
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retention strengths  and  grouping tendencies,  the DNAP-ailica was  found to  be
superior than alumina and silica-R (NH ) .
     A  combination  of  liquid  chromatography   (silica  gel column)  and GC-FID
method was  employed  by Fett  et al.  (1968)  routinely to  determine  toluene  in
hydrocarbon solvents.
8.5.  BIOLOGICAL SAMPLES
   \  toluene or its metabolites' have been determined  in blood, in urine, and  in
mothers' milk samples.  These methods of analysis are discussed  below.
8.5.1.  Blood.  Toluene in blood has  been determined by  GC analysis of headspace
samples (Premel-Cabie et al.,  197*1; Anthony  et  al.t  1978;  Radzikowska-Kintzi and
Jakubowski,  198U.  According to this method, blood  is equilibriated  with air  in
a closed container at a fixed  temperature.   The  headspace  gas  is  injected into  a
GC-FID  system  for  detection of toluene.  The  method can be  used for quantifi-
cation  of  coluene in  blood by  the  standard  addition  method as  described  in
Section 8.2.2.2.
8.5.2.  Uriny.   In  the body,  toluene is mainly oxidized  to benzoic  acid which,
after conjugation with glycine, is  eliminated  as hippuric acid in  the urine.
Hippuric acid may be formed from other  metabolic processes besides toluene meta-
bolism.
     Hippuric acid in  urine can be  determined by a number of methods including
colorimetry  (Uraberger  and  Fioresse,   1963),   UV  spectrometry  (Pagnatto  and
Lieberman,  1967),  and thin-layer  chromatography (Bieniek  and  Wilczok, 1981);
however,  one of  the  better  methods of  hippuric acid  analysis in urine was
developed  by Caperos  and  Fernandez  (1977).   According to this  method,  the
hippuric acid in acidified urine is extracted with ethyl acetate.  The extracted
hippunc acid is esterfied. with 1-p_-tolyltriazene.  The  dried  ester is dissolved
in  chloroform and quantified  by GC-FID.   The recovery of hippuric acid  by  this
method  is  determined  from the  recovery  of an  added internal  standard.   The
sensitivity  of  the method with  0.5 ml urine  was determined to be 5 ppm.
     Hippuric acid is an endogenous metabolite  common  in human urine,  but toluene
exposure enhances  jts level.    However,  o-cresol  may be  a  more specific urine
metabolite and  may be  regarded  as  a  better  index of  toluene  exposure in humans
(Hansen and  Dossing,  1982).   A  recent  method (Hansen and Dossing, 1982) deter-
mined the urinary hippuric acid and o-cresol levels  by a high-porfcrmance liquid
chromatographic ( HPLC) method.  In this method, the  hippuric acid level  in urine
was determined  by extracting it with  acetonitrile and injecting  the extract  onto
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the HPLC column.  The o-cresol  level  in urine was  determined  by digesting the
urine with  concentrated sulfuric  acid and  extracting the  digest  with cyclo-
hexane.   The cyhclohexane  layer was first  washed with a  phosphate  buffer and
finally extracted with sodium  hydroxide.  The aoiiooi ii/u« uAiut p'uaac wds  injected
onto the HPLC column for the determination of o-cresol level.  The HPLC system in
both  cases  consisted of  a Lichrosorb Si  60  column  and  a  UV  detector.   The
detection limits were found to 0.05 rag/mi  and 0.05 ^g/mi for  urine hippuric acid
and £-cresol,  respectively.
8.5.3-    Mother's Milk.  The  levels of toluene in mother's milk  for populations
in the  vicinity  of chemical manufacuring  plants and/or industrial user  facili-
ties  in the United  States were  Measured  by Pellizyari  et al. (1982).   The
volatile compounds including toluene in the  milk  samples were determined by the
purge and  trap  method  (Section 8.2.2.1.),   followed  by thermal  desorption and
capillary GC-MS analysis.    Of the  total of  12  samples   collected,  8   samples
qualitatively showed detectable levels of  toluene. The detection limit for these
analyses was not specified by the authors.
8.6.  FOODS
      A  headspace GC technique for quantification  arid  a GC-MS technique  for con-
firmation were  used to determine trace amounts of toluene  in plastic containers
(Hollifield et  al., 1980).  The sample, taken  in  a specially enclosed vial, was
heated  at 90°C for 2 hours, and 2  mi.  of  headspace gas was  injected  into  a GC
system.  The principle  of  standard addition was  used  for  the quantification of
toluene.  Toluene present  in  parts  p£~ billion range can   be determined  by this
nuthod.
8.7.  CIGARETTE  SMOKE
      The concentration of toluene both in  sidestream smoke  (Jermini et al.,  1976)
and  mainstream  smoke (Dalhamn  et   al.,  1968a) has  been   determined.    For the
determination of toluene in mainstream smoke, standard cigarettes w?re smoked by
machine under standardized conditions (a 2 second  35 mi. puff  once every  minute).
The mainstream  smoke  is  collected  in a cold trap (DaJhamn et al., 1968b).  The
contents of the  cold trap  can be introduced into  the  GC by multiport valves and
analyzed by GC-FID for toluene  determination.
      Toluene determination in  sidestream  smoke can be accomplished by  adopting
the  sampling  and analysis  technique  of Holzer et  al. (1976).    The sidestream
smoke can be collected by  drawing  the  smoke through a solid  sorbent tube packed
with  Tenax GC.   The  Tenax  GC  sorbent tube  can be thermally  eluted onto a  glass
                                      8-13

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capillary column for the determination of  toluene  content.   Adoption of a cojd
trap for solitless injection of  the  sample into the capillary column (Grob and
Grob technique)  will enhance the sensitivity and accuracy of the method.  Addi-
tional confirmation of the GC peaks can be done by interfacing the GC with a MS
(Holzer et al.,  1976),

8.8. REFERENCES

ALTSHULLER,   A.P.,   LONNEMAN,   W.A.,   SUTTERFIELD,   F.D.   and  KOPCZYNSKI,  S.L.
(1971).    Hydrocarbon  composition  of  the  atmosphere  of  the  Los  Angeles
Basin—1967.   Environ. Sci. Tech.  5: 1009.

ANTHONY, R.M., BOST,  R.O., THOMPSON, W.L. and SUNSHINE,  I.  (1978).  Paraldehyde,
toluene, and methylene  chloride  analysis by headspace  gas  chromatography.   ^J.
Anal. Toxicol.   2: 262-2614.

BALL, H.  (1976).   Some  new  aspects in air pollutants analysis  of hydrocarbons by
automatic gas-chromatography.  Fresenius Z. Anal. Chea. 282: 301-305.

BELLAR, T.A., BUDDE,  W.L. and  EICHELBERGER,  J.W.   .(1979).  The indentification
and measurement  cf volatile organic compounds in aqueous environmental samples.
In:  Monitoring Toxic Substances.  ACS Symposium Series, p. 49-62.

BELLAR, 7. A.  and LICHTENBERG,  J.J.  (1979).  Semiautomated headspace analysis of
drinking waters  and industrial waters for purgeable volatile organic compounds.
In:  Measurement of  Organic  Pollutants in Water and Vastewater,  ASTM STP 686.
Van  Hall,  C.E.,  ed.   Philadelphia,  PA:    American  Society  for  Testing  and
Materials, p. 108-129.

BERTSCH, W,,  ANDERSON,  E. and HOLZER,  G.   (1975).  Trace  analysis of organic
volatiles in  water by gas chromatography-mass spectrometry with glass capillary
columns.  _J.  Chromatogr.  JJ£: 701-718.

BIENIEK, G. and  WILCZOK, T.   1981.  Thin-layer  chromatography of hippuric  and m-
methylhippuric  acid in urine after mixed exposure to toluene and xylene.  Brit.
i' -Ind- Med.   38:- 304-306.
                                      8-11

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BRODOWSKI,  P.T. ,  WILSON,  N.B.  and SCOTT,  W.J.   (1976).   Chromatographic  analysis
of gaseous  products from  pyrolysis or organic  wastes  with 'a. single column.  Ana 1 .
Chem.   18(1?) :  1812-1813.

BURGHARBT,  E. and JELTES,  R.    (1975).   Gas Chromatographic determination of
aromatic hydrocarbons  in air  using a semi-automatic  preconcentration  method.
       Environ.   9_: 935-9140.
CAPEROS,  J.R.  and FERNANDEZ,  J.G.  (1977).  Simultaneous determination of toluene
and xylene metabolites in urine  by gas  chromatography.   Brit. _J.   Ind.  H^d .
31: 229-233.

CHOVIN, P. and LEBBE, J.  (1967).   Chromatography of  aromatic hydrocarbons.   I.
The .determination of gas  chromatogr&phy of aromatic hydrocarbons in  the  air  of
    !
working environments.  Occup. Health Rev.   19( 1-2) : 3-10.

DALHAHN,  T. ,  EDFORS,  M.L.  and RYLANPER,  R.  (1968a).  Mouth absorption of various
compounds in cigarette smoke.  Arch. Environ.  Health.   16(6) : 831-835.

DALHAHN,   T.,  EDFORS,  M.L. and RYLANDER,  R.    (1968b).   Retention of cigarette
scvoke components in human lungs.  A_rcf_.  Environ. Health.   17 : 7^6-7^8.

DeVERA, E. R. ,  B. P. SIMMONS, R.D. STEPHENS, and D.L.  STORM.  (1980).   Samples and
Sampling Procedures  for Hazardous Waste Streams.   U.S.  EPA  Report No.    600/2-
80/018.  Municipal Environmental Research Laboratory,  Cincinnati,  OH. Available
through NTIS.   Order No.  P3 80-135353,  Springfield,  VA.

DOWTY, B.J.,  ANTOINE, S. R. and LASETER, J.L.  M979).   Q'-sr.t i tat, i vc  zr,Z  qualita-
tive analysis of  purge*bli organics by high-resolution gas chromatography and
flase ionization detection.  In:  Measurement of Organic Pollutants  in Water and
Wastevater.  ASTM  STP 686.   Van Hall,  C.W.,  ed.   Philadelphia,  PA:   American
Society for Testing and Materials,  p.  24-35.
                                      8-15

-------
DKOZD,  J., NOVAK,  J.  and  RIJKS,  J.A.   (1978).    Quantitative and  qualitative*
headspace gas analysis of parts per billion amounts of hydrocarbons in water.   A
study of  model  systems by  capillary-column gas  chromatography with  splitless
sample injection.  _J.  Chromatogr.   156:
ESPOSITO, G.S.  and  JACOBS,  B.W.   (1977).   Chromatographic  determination  of
aromatic hydrocarbons in ambient air.  Amer.  Ind.  Hy_£.  Assoc.   36 :  H01-H07.

FEDERAL REGISTER.  (1979).  Purgeable Aroraatics— Method  602.   Federal  Register.
FETT,  E.R.,  CHRISTOKFERSEN,  D. j .  and SNYDER, L.R.  (1968).  Routine determination
of benzene,  toluene,  ethyibenzene and  total aromatics in hydrocarbon  solvents  by
a combination of liquid and  gas chromatography .   _J.  Gas  Chromatogr.   6_: 572-576.

FRACCHIA, M.,  PIERCE,  L. ,   GRAUL,  R.  and  STANLEY,  R.   (1977).   Desorption  of
organic  solvents  from charcoal  collection tubes.    Amer.  Ind.  J^. Assoc   _J_
PHASER, D.A and RAPPA'PORT,  S.   (1976).   Health aspects of the curing of synthetic
rubbers.  Environ. Health Perspect.  17 : ^5-53.

GRIZZLE, P.L. and J.S.  THOMSON.   (1982).  Liquid Chromatographic  separation of
aromatic hydrocarbons with chemically bonded  ( 2, U-dinitroanilinopropyl) Silica.
Anal. Chem.  5J*_: 1071-1078.

GROB, K. and  ZURCHER,  F.  (1976).   Stripping of trace organic substances  from
water:  Equipment and procedure  _J. Chromatogr.   117 •' 285-29^.

GROB, K. and  GROB,  G.    (1971).   Gas-liquid  chromatographic/inass  spectrometric
investigat-.on of C.f-C-n  organic compounds in an urban atmosphere.   J.  Chromatogr.
62_: 1-13.   (Cited in Syracuse Research  Corporation,  1980).

HANSEN, S.H.  and DOSSING, M.  (1Q82).  Determination of urinary hippuric acid and
o-cresol,  as  indices  of toluene exposure,  by liquid chromatography  on dynami-
cally modified silica.   J. Chromatogr.   229:  111-1H8.
                                      8-16

-------
HESTER,  N.E.  and MEYER,  R.A.  (1979).  A sensitive technique  for measurement  of
benzene  and  alkylbenzenes in air.  Envi ron. Sci. Technol.   1j(1) ; 107-109.

HOLLIFIELD,  H.C..  BREDSH,  C.V.,  DENNISON, J.L.,  ROACH,  J.A.  and  ADAMS,  W.S.
(1980).   Container-derived  contamination  of  maple  syrup with methyl  raethacry-
late,  toluene,  and  styrene as  determined  by headspaee  gas-liquid  chromatography.
J_. Assoc.  Off.  Anal.  Chen.   63.: 173-177.

HOLZER,  G.,  ORO, J. and BERTSCH, W.  (1976).  Gas chromatographic-maas spectro-
aetric evaluation of exhaled tobacco smoke.   J.  Chrooaatogr.   '\ 26 : 771-185.

HOLZER,  G.,  CHANFIELD,  H.,  ZLATKIS,  A.,  BERTSCH,  W., JUAREZ, P.,  HAYFIELD,  H.
and L.IEBICH,  H.M.  (1977).   Collection and analysis  of  trace  organic  emissions
from natural sources.  J. Chromatogr.   1*42: 755-764.

ISIDOROV,  V.A., ZEKKEVICK,   I.G.  and  IOFFE, B.V.  (197-7).   Investigation  of new
sorbents for the gas-chromatographic-ciass-spectrooetric  determination  of  traces
of  volatile  organic  compounds  in the  atmosphere.   Translated froc Doiclady
A'tCa^gmii NauK iO5_K,  
-------
JUNGCLAUS,  G.A., LOPEZ-AVILA, V. and KITES,  R.A.   (1978).   Organic  ccapounds  in
an industrial wastevater:  A case study of their environmental impact.   Envi ron.
Sci.  Technol.  12(1); 88-96.

KRPST,  K.J.,  PELLIZZARI, E.D., WALBURN, S.G. and HUBBARD, S. A.  (1982).   Collec-
tion and analysis of hazardous organic  emissions.   An a . 1 .  Chgm .   51 :  810-817.

LINGC,   R.D.,  MELTON,  R.G.;  KOPFLER,  F.C., COLEMAN,  W.E. and  MITCHELL,  D.E.
(1977).  Quantitative analysis of volatile organic compounds by GC-MS.  J_.  Areer.
Water- Works Assoc.-  69(11, pt . 1): 605-61?.

LONSEMAN, W.A.,  BELLAR,  T.A.  und  ALTSHULLER,  A. P.  (1968).  Aromatic  hydrocarbons
in  the  atmosphere  of  the  Los  Angeles  Basin.     Erv: ron.   Sci .   Technol.
2(1 M:  1017-1020.

LOUW, C.W.  and RICHARDS, J.F.  (197b'.   A simple directly combined gas chromato-
                                            «w
graphic-infrared  spectrometri c  systen  for indentif ication  of  low molecular
weight  hydrocarbons.  App J .  Spec t rose.   2_9: 15-2*1.

MINISTRY OF LABOUR.  (1966).  Methods  for the Detection  of Tox.c Substances  in
Air, POO k let No.  u :   'oen?en^,  Toluene  and  Xylene,   Styrene.   London:   Her
Majesty's Stationery Office, pp.  1-12.

NELlGAfi, R.E. ,  LEOSAM;,  M.J.  and BR1TAN, R.J.   (1965).   The gas  chromatographic
detersalr.atitin of aromatic  hydrocarbons in  the ataosphere.   Reprint  of  paper
presented to the Division of Water,  Air, Lnd Waste  Chemistry,  American  Chemical
Society, Atlantic City.  N.J,  Septemfce-  12-'' 7,  1965,  2 p.
     , P.K. and  FISHBURN,  P. J,   '1977).   The characteristics Of  odours  by  gas
chrooatography.  In:   Analytical  Techniques in the Determination  nf  Air  Pollu-
tants:  A SynposiuE.  Clear  Air Society of  Australia  and  New  Zealand,  p.
                                      8-18

-------
NIOSH (NATIONAL  INSTITUTE FOfi OCCUPATIONAL SAFETY AND HEALTH).  (1977).   Toluene.
In:   NIOSH Manual  of Analytical Methods,  >nd edition. Part II.  Standards Comple-
tion Prograa Validated Methods, Vol. 3.  NIOSH Publication No.  77-157-C,  p.  3"3-1
to 313-6.   U.S.  Dept.  of Health, Education, and Welfare,  Public  Health  Service,
Center for Disease Control, NIOSH, Cincinnati, OH.

NRC  (National Research  Council).   (I960).   The Alkyl  Benzenes.  Committee  on
Alkyl Benzene  Derivatives,  Board  on Toxicology  and  Environmental  Health  Hazards;
Assembly of Life Sciences,  National Research Council.  Hashinfton,  DC:   National
Acadeay Press.

OGATA, M.,  A3AKARA,  H.  and SAEKI,  T.   (1975).  Sampling and analysis  of  some
aromatic,  aliphatic and  chlorinated hydrocarbon vapours  in  air:  A gas-liquid
chromatographic ar.d colorimetric method.  I^nt. Arch. Art>eitsmed.   j1*: 25-37.

PAGNOTTO,  L.D. and LIE8ERMAN, L.M.  (1967).  Urinary hippuric acid excretion  as
an index of toluene exposure.  Aaer.  Ind. Hy_£. Assoc. ^.   28: 129-131*.

PELLIZZARI, E.D., BUNCH, J.E., BERKLEY,  R.E. and MCRAE,  J.   ('.976).  Determina-
tion of trace hazardous  organic  vapor pollutants in ambient  atmospheres by gas
chromatography/aass speetroeetry/computer.  Anal.  Chen.   48:  803-807.

PELLIZZARI, E.D.,  HARTWELL,  T.D.,  HARRIS,  B.S.,  WADDELL, R.D., WHITAKER,  D.A.
and ERICKSON,  M.D.  (1982).  Purgeable organics in mother's milk.  Bull.  Environ.
Contaa.  Toxieol.  28: 322-328.

PFAE.VDER,  F.K.  (1976).   Analytical Methods Developed.   E5E  Notes.   12(j|) :  4-5.

PILAR, S.  and  GRAYDON, W.F.  (1973).  Benzene and  toluene distribution in Toronto
atmosphere.  Envlron.  Sci.  Technol.  7(7) : 628-631.

PREMEL-CABIC,  A.,  CAILLEUX,  A.  and  ALLAIN,  P.   (197^4).   [Identification and
quantification by gas  chromatography of  fifteen organic  solvents in  the blood.]
Clin. Chim. Acata.  56:  5-11.  (In Fr.).
                                      8-19

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RADZIKOWSKA-KINTZi,  H,  and JAKUBOWSKI, M.  (1981).   Internal  standardization  in
the head  space  analysis  of organic solvent  in  blood.   Int. Arch.  Occup.  Er.yj, ron.
Health.   ^9:  115-123.

RAWLINGS, G.C. and  SAMFIELD, M.   (1979).    Textile plant wastewater  toxicity.
Environ.  Sci.  Technol.   13(2): 160-161.
REID, F.H.  and HALPIN,  W.R.  (1966).  [determination of  halogenated  and  aromatic
hydrocarbons in air by charcoal  tube and gas  chrcxaatography.   Amer. Ind.  H.y_£.
Assoc.  J.   29 ( H):  390-396.

RYAN, J.P.  and FRITZ, J.S.   (1978).  Determination of trace organic impurities in
water using thermal desorption by XAD resin.   J. Chromatogr. Sci.   16:  U88-492.
SCHNEIDER,  W.,  FROHNE, J.C. and- BRUDERRECK, H.  (1978).   Determination of hydro-
                            q
carbons in the  parts  per 10  range  using glai
aluminun oxide.  J. Chromatogr.  155: 311-327.
                            q
carbons in the parts  per  10  range  using  glass capillary  columns  coated with
SEIFERT,  B. and ULLRICH,  D.   (1978).  Determination  of  organic  pollutants  by  gas
chromatography after cryogenic sampling.  Stud.  Environ.  £>_cl_.  J_:  69-72.

TOKUNAGA,  R.,  TAKAHATA,  S.,  ONODA, M.,  ISHI-I,  T., SATO,  K., HAYASHI, M.  and
IKEDA, M.   (197^).   Evaluation  of the  exposure   to  organic  solvent  mixture.
Comparative studies on  detection tube and  gas-liquid  chroraatographic  methods,
personal and  stationary  sampling, and urinary  metabolite determination,   Int.
Arch. Arbeitsmed.  3J[: 257-267.

TWIBELL,  J.D.  and HOME,  J.M.    (1977).    Novel  method  for  direct analysis  of
hydrocarbons  in  crime   investigation  and  air pollution  studies.     Nature.
268: 711-713.

UMBERGER, J.C. and FIORESSE,  F.F.  (1963).  Colormetric method  for hippuric acd.
Clln. Chem.  1: 91-96.
                                      8-20

-------
U.S.  EPA (U.S.  ENVIRONMENTAL PROTECTION  AGENCY).   (1979).   Chemistry Laboratory
Manual for Bottom Sediments and Elutriate Testing.  U.S.  Environmental  Protec-
tion   Agency.  Chicago,  J.L.   Available  from:   National  Technical  Information
Service, Springfield, VA  (NTIS  PB 294-596).
     \
VAN ERT, M.D.,  ARP,  E.W.,  HARRIS,  R.L.,  SYMONS, M.J. and WILLIAMS, T.M.  (1930).
Worker exposures  to  chemical agents in tne manufacture  of rubber tires:   Solvent,
vapor studies.  Amer.  Ind. Hyg.  Assoc.  J.   *n :  212-219.

VITENBERG, A.G.,  STQLYAROV,  B. V. and  SKIRNOTA,  S.A.    (1977).   Gas-chromato-
graphic determination of traces of aromatic hydrocarbons and alcohols in water by
the  equilibriun  vapor  analysis  method.   Vestn.  Leningr.  Univ.,  Fiz.  Khlm.
3: 132-139.
                                     8-21

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                            9.   EXPOSED POPULATIONS

     Vhe  number  of people exposed to various  sources  of toluene can be divided
into three categories,  general  population,  occupational group,  and  cigarette
smokers.   The breakdown of general population subjected to inhalation exposure of
toluene  from  various  sources of  emissions can be obtained by performing a popu-
lation analysis around  each source.   A computer program was  used  by Anderson
et al. (1980) to extract site-specific  population patterns from the U.S.  Census
figures  standardized  to  1978 population levels.  The number of general population
exposed  to various  levels  of toluene   from  different  sources  of  emission  as
calculated by  Anderson et  al.  (1980) is shown in Table 9-1.   For an explanation
of the breakdown of  the source variety  shown in Table 9-1,  see Section 10.1.1.
     The exposed population count shown in Table  9-1  is derived  from the geo-
graphical coordinate  of  each location.  Error in the geographical coordinates of
a source and  population center will  cause errors in population count.   In addi-
tion, the population  count  figures  obtained  from the  U.S. Census Bureau  are
subject   to  undercounting.   The  result of  this  undercounting  will   be  lower
population exposure  estimates  than the  actual case.
     No  estimate of  the number  of  general population  exposed  to toluene from
ingestion of  foods  and drinking  waters  can  be given.   Toluene has been detected
in only  a small  fraction  of total  drinking water  supplies  and  foods  that have
been monitored.  The number of people consuming the contaminated waters and foods
is not known  at  the  present  time.
     The three most likely sources that may lead to dermal exposure of toluene to
the g'.?neral population  are  usage of vehicular fuels,  toluene-containing sol-
vents, and cosmetic  products containing toluene.  With  the recent  increase of
self-service  gasoline stations around the country, the number of people who may
inadvertently spill gasoline on  parts  of their  body  during filling operations
must  have increased  by a large  number.   The  deliberate  use of solvents  for
cleaning body grease or  inadvertent  spillage  of  cleaning solvents  and  paint
thinners on parts  of the body  will a]so  lead  to dermal exposure  to toluene.
Although the  extent  of exposure  may  be  much  less  significant compared to the two
aforementioned   sources,   users  of  cosmetic  products  containing toluene  are
another  group of the general population  that  is exposed to toluene through the
dermal route.   However,  no estimate  is  available on the number of the general
                                      9-1

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                                    TABLE  9-1
                 Population  Distribution  and  Inhalation Exposure
                    Levels of  Toluene  from Different Sources3
Concentration
Level
>100
100 - >50
50 - >25
25 - >10
10. - >5
5 - >2.5
2.5 - >1
1 - >0.5
0.5 - >0.25
0.25 - >0.1
0.1 - 0
Subtotals
Total

Specific
Point Sources
0
0
34
475
1,^34
6,103
19,781
39,064
95,883
269,883
34,316,299
34,748,633
195,
Number of People Exposed
Prototype
Point Sources
'59
2,841
10,200
22,700
33,900
75,200
240,000
246,000
350,000
1,229,000
0
2,210,000
637,768
From
Area
Sources
58,347
446,793
12,3^8,504
42,478,913
66,368,769
0
0
0
0
0
34,977,809
158,679,135

Source:   Anderson  et  al.,  1980
                                        9-2

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population dermally exposed to toluene from  these  sources.   The usage of other
consumer  product  formulations containing toluene  (see Table  4-20)  may cause
inhalation/dermal exposure  to toluene.    An  estimate  of  the number  of people
exposed to toluene from these products is also unavailable.
     According to  the  estimate  of  the  Department  of Health,  Education,  and
Welfare (1977), more than  4.8 million people  per  year are occupationally exposed
to toluene.  Toluene ranks fourth among all other agents listed in terms of the
number of people exposed to any single agent.
     The number of people  in the  U.S.  exposed to  toluene through cigarette smoke
has been  estinated  to be  56 million  during  the  year 1978a.  This figure which
considers the exposure to the smokers only, is bound to  be an underestimate since
it does not include passive smokers.

9.1. REFERENCES

ANDERSON, G.2.,  LIU, C.3.,  KOLMAN,  M.Y. and KILLUS,  J.P.  (1980).  Human Exposure
to Atmospheric Concentrations  of Selected Chemicals,  Publication No. unavail-
able.  Prepared by Systems Applications,  Inc., San Rafael, CA,  under Contract No.
EPA 68-02-3066.   U.S.  Environmental Protection Agency, Research Triangle Park,
NC.

PUBLIC HEALTH SERVICE.  (I930).  Smoking, Tobacco end Health,  A Fact Book.  U.S.
Dept. of Health and Human  Services,  Public Health Service,  Office on Smoking and
Health.

U.S. DEPARTMENT  OF  HEALTH,  EDUCATION, AND WELFARE.   (1977).    National Occupa-
tional Hazard Survey,  Vol.  III.  Survey Analysis and Supplemental Tables.  U.S.
Dept.   of Health,  Education,  and  Welfare,  National  Institute of Occupational
Safety arid Health, Div. of  Surveillance,  Hazard  Evaluations,  and Field Studies,
Cincinnati, OH, p. 448.
 o
  This figure is based on the following assumptions:  Of the total  population  of
 225  million,  21.4$ are  under  age 13  (Dept.  Commer.,  1979)  and  do not  smoke.
 Teenagers in the age group 13 to 17 years  constitute 7.6% of the total  population
 (Dept. Crmmer., 1979).   Of the  7.6%  of the  teenagers,  only  11.7? are  assumed  to
 be  smokers  (PHS,   1980).   Of the  remaining population, 51$  are  assumed to  be
 females and 49$ to be males  (Dept. Commer., 1979).   The percent of  female and
 male smokers over  age 17 are assumed to  be 30.4$  and  37.4$,  respectively (P!"1,
 1980).
                                       9-3

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                        10.   ESTIMATE OF HUMAN EXPOSURE

     Exposure is the contact between a subject of concern  and an agent such as a
chemical,  biological, or  physical entity.   The  magnitude  of the  exposure  is
determined by measuring or  estimating the amount of  an  agent  available at the
exchange boundaries, that is, lung,  gut,  and  akin,  during some specified time.
Exposure assessment is the qualitative estimation or quantitative determination
of the magnitude, frequency, duration, and route of exposure.  Exposure estimates
are often combined  with  environmental and health effects data in performing risk
assessments.   The  exposure  of an agent  may  lead to the  intake of  some of the
agent.  Uptake of an absorbed dose is the  amount  of the intake that  is absorbed by
the subject.
     The assessment  of  human health  risks  from  exposure to  any  environmental
pollutant requires  knowledge of (1) the dosage of the pollutant received by the
exposed human population  and (2) the effect  of the pollutant  on  human health.
Because it is not the  purpose of  this  section to develop a health effects model,
no attempt will  be  made  to address such parameters as population characteristics
(e.g., age, sex, occupation,  racial  background),  population habits  (e.g.,  food
habits,  recreational habits,  product-use  habits),  and  population  groupings
(e.g.,  the  aged,   pregnant  women,  children,  other  high  health  risk  groups).
Instead,  this section will  attempt  to  derive  the human  exposure  of toluene
received from all sources of emissions.
     To estimate human exposure, one must consider  route of entry, magnitude of
exposure,  frequency  of  exposure,   and  duration  of  exposure.    The  general
population  may- be exposed  to  toluene  through the  following  routes:    (1)
inhalation of air,   (2)  ingestion of water and  foods,  and (3)  exposure through
skin.  The next  step combines the estimation of environmental concentrations with
a description of the exposed population to yield  exposure profiles and exposure
pathway analysis.
     Certain  segments of  population may be exposed to  toluene through occupa-
tional exposure  and cigarette smoking.   Because exposure  of this  segment falls
under a special category, these scenarios will  be  discussed separately.  This
section does  not include toluene  exposure from the use of consumer products.  As
has  been  mentioned  in  Subsection 10.5.,  some consumer  products  contain  high
percentages of toluene.
                                      10-1

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Undoubtedly,  the  use of  these  consumer  products  leads to  various  degrees of
toluene exposure in the general population;   however,  no data are available to
derive  estimates   of toluene  exposure  frote  consumer  products.    Also,  the
conversion factors  for  expressing toluene  concentrations  in air  at  25°C are:
1 ppm :.  3-77  rag/m-5 and in water.,  1 ppm :.  1 mg/t.
10;1.  EXPOSURE VIA INHALATION
   \  Toluene  exposure via inhalation  can  be estimated  in two ways.   First, the
exposure can  be estimated  from the  total nationwide toluene emission data by the
use  of  mathematical models  simulated  to  reflect  the actual  environmental
setting.  Second,  the  exposure  can  be  estimated from  actual  monitoring data.
Estimating e^osure on the basis  of monitoring data  is  often  a preferred method,
because these data  directly provide  the  environmental  distribution of toluene;
however, this method lias limitations.  Although the monitoring data available for
toluene are more abundant  than those  available for many  other organic  chemicals,
they may  not  be statistically representative  of  all  the population  exposed to
toluene.   The monitoring  data  may  not  provide  information on the  extent  of
concentration  variation  due  to  chemical   reactivity  (e.g.,  photoreaction,
oxidation in  the atmosphere, etc.).  These  data also do  not  yield relationships
between  materials  balance  of  the  emitted  toluene   and  the  environmental
concentration distribution in an area.  Therefore, the approach toward exposure
estimation in this  section  has used  both the available  ambient monitoring data
and the theoretical dispersion modeling of  toluene emission  data.
10.1.1.  Theoretical Modeling.  The estimation of  inhalation  exposure  to toluene
among different segments  of the general population involves the  following compu-
tational  tasks:   (1) estimation  of annual average toluene concentration in the
air at different distances  from the  emission sources  and (2) estimation of the
population distribution  around  each source  of emission (available through the
U.S.  Census  Bureau).  The latter  computation has  already  been  discussed  in
Chapter 9.
     The performance of the first  task requires the following data: (1) emission
inventories of toluene, which are already available (Subsections ^.^.l. through
4.^.4.),  (2)  atmospheric  reactivity  of  toluene,  (3) meteorological data, which
are available  through  the U.S.  or local  weather  bureau, and  (1)  a   dispersion
equation to estimate concentration distribution of toluene.
                                      10-2

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     Toluene concentration  downwind from a  source  can be  estimated  using the
following dispersion equation (Turner, 1969):
                     C(X,0,0) =   .  Q  •
exp
                                                    2a
                                                      z
where

     C(X,0,0) - concentration of toluene at various x coordinates and at zero y
                and z coordinates (mg/m )
            Q = emission rate (mg/s)
           o  - horizontal dispersion coefficient of the plume concentration
                distribution
           a  _ vertical dispersion coefficient of the plume concentration
                distribution
            U = the mean wind speed (m/s) (w = the heat of the source)
            h - the effective stack height; i.e.,  the sum of the stack height  and
                plume rise (m)
     Assuming U =  5 m/s; Q  =  200 x. 10  kg/year - 6.31 x 10^ mg/s; plume height
= 10 m  and  20 m;  and  the  values  of a  and  o   from the  following equation
(Anderson et al.,  1980):
               O   (m)   0.06x(1 H- 0.0015x)~1/2
                z                          1 /•>
               O   (m) = O.OBxd + O.OOOIxT
one can calculate  the concentration of  toluene at different distances from the
source, as given in Table 10-1.
                                      10-3

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                                   TABLE 10-1

          Concentration  of  Toluene  (mg/nr)  at  Different  Distances  (m)
               From  A  Source Emitting  200  Million  kg/Year Toluene3
Plume Height
(m)
10
20

100 500 1,000 1,500
1.36 0.45 0.15 0.12
0.003 0.31 0.13 0.1C

5,000 10,000
0.02 0.01
0.02 0.01
Source:   Slimak,  1980

       The  calculations  of  the values in Tab3e 10-1  for  toluene distribution from a
  stationary  source do  not consider the  chemical  reactivity  of  toluene in  the
  atmosphere  and  the effect of plume temperature on the concentration distribution
  of toluene.   A  more  detailed  calculation thst incorporates these two variables,
  as well as  building  wake  effect  (enhanced  dispersion  due to buildings), has been
  made for the  estimation of  spatial  concenr.ration  of tcluer.e  from  the  najor
  stationary  sources  of  toluene  emission  (Anderson  et al.,  I960).
       The  dispersion equation  developed  by Anderson  et  al.  (1980) was  used  to
  compute annual  average concentration  pattern of toluene from each point source.
  A computer  program  was used to  evaluate  these concentration patterns from  the
  given  meteorological and emission data.   Because  there  are  numerous sources of
  emission,  the sources  were divided into  three types, each of which  is defined
  below.
            Specific  Point Sources:  These  sources  were treated  using parameters
       appropriate to  each  source.  Included are emissions from production sources
       and  from chemical intermediate  users.
           General Point  Sources:  For  these sources, a prototype analysis  was
       done and the results were  multiplied by the estimated  number  of sources.
       Thes-,'  sources   included emissions  from gasoline  marketing,  from the  coke-
                                        10-1)

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     oven  industry,  and from  isolated  and non-isolated toluene producers  (not
     included  in the  previous  categories).
           Area Sources:  Such sources were treated as emission per unit area  over-
     identified  areas.   These sources  included  mobile emission, emission  from
     solvent use, and emissions  from miscellaneous sources4

;     The  three equations used to calculate the spatial concentration  distribu-
 tion of toluene from ail  sources  are given in considerable detail  in  Anderson
 et  al.  (1980); interested  readers  are referred  to that document.   The final
 results  of the  calculations  of Anderson  et al.  (1980)  led to the  estimate  of
 apatial  concentration range of  toluene  around different sources of  emissions.
 These  values are given  in  Table  10-2.

                                     TABLE 10-2
                  Population  Distribution and  Inhalation Exposure
 '                     Levels of Toluene From Different  Sources
Number of People Exposed From
Concentration
Level
(ug/m3)
>100
100 to >50
50 to >25
25 to >10
10 to >5
5 to >2.5
2.5 to >1
1 to >0.5
0.5 to >0.25
0.25 to >0.1
0.1 to 0
Subtotals
Total
Source : Anderson
Specific
Point Sources

0
0
34
475
1,13"
6,103
19,781
39,064
95 , 560
269,883
34,316,299
3t.7t8.633

et al., 1980
Prototype
Point Sources

159
2,841
10,200
22,700
33,900
75,200
2tO,000
246,000
350,000
1,229,000
0
2,210,000
195,637,768

Are?
Sources

58,3t7
446,793
1 2, 348, 504
42,478,913
68,368,769
0
0

L
0
34,977,809
158,679,135


                                         10-5

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     Anderson  et  al.  (1980)  listed the following factors that could cause uncer-
tainties  in their calculated exposure levels given in Table 10-2:
     Emission  Estimates Errors:   Some of these  are  (1) error in the estimates of
     production and use of toluene,  (2) the  assumption that all plants operate at
     the  same  capacity,  (3)  omission of  certain emission sources, and CO  error
     in derivation of  emission  factors  and,  in  certain  cases,  the  use  of  a
     uniform emission factor,  which implies 'that  all these plants have similar
     emission  controls.   It  is difficult to  project  whether  the emission  esti-
     mates used  by Anderson et al.  (1980) will  lead to higher or lower exposure
     estimates.   This can be done,  however,  by comparing  these estimates with
     the  experimentally determined  concentration patterns obtained from sources
                                                                          V
     that are  reasonably isolated from other sources.
     Concentration Pattern  Errors:   The  concentration patterns  used in  the
     exposure  computations were obtained througn atmospheric  dispersion model-
     ing.  Any deviations  in these estimates from the  true pattern (difference in
     theoretical  ar,d  experimental values) directly affect the exposure results.
     Many assumptions were  used  in  calculating the concentration distribution.
     The  exposure errors  will be more  severe  in  the  case of  prototype  point
     sources where a  prototype model was used for  calculating exposure from  all
     other similar sources.   The same can  be said  about  the  exposure estimates
     from area sources  based on  a box model method  that incorporated a number of
     uncertainties.
          Interpolation Errors:   The interpolation of population and concentra-
     tion patterns used to develop  patterns of  exposure can introduce errors.

     With the available  information, it is  impossible to quantify any of  the
errors described  above.   The theoretical model  may provide  qualitative insights
in certain instances  to predict  whether  the exposure estimate is either too high
or too low compared  to  the actual values.
10.1.2.  Inhalation  Exposure Based  on Monitoring Data.   Exposure of the general
population to toluene by  inhalation can occur under  a wide range  cf exposure
scenarios.  Because  it  may be considered impractical to measure toluene concen-
tration from all possible exposure scenarios, an atteipt has been made to develop
a few of  the most prevalent ones.
                                      10-6

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     The  four  largest sources  of  toluene  emission,  in  descending order,  are
automobile use  (exhaust  emission, engine evaporative loss, gasoline  marketing
evaporative loss),  Industry  sites using toluene as  a  solvent,  coke  oven  sites,
and toluene production sites (Subsection l.i|.H.).  in place of  dispersion  model-
Ing, one can use the monitoring data from each of the four sites to evaluate the
four different  exposure scenarios. The a./Ticulty with this approach is that the
available monitoring data were often developed for sines with various degrees ot
intermixing between  these  exposure scenarios.   Therefore,  inhalation  exposure
has been classified under three scenarios: the urban areas, areas containing the
user sites, and ru^al  or reaote  areas.   In this manner,  the exposure  estimates
developed  may   be  representative  of a  broad  range of  the possible  exposure
scenarios.  It should  b-j re«eaoered that  the  urban  areas  may contain sites  with
high automobile use,  production  and other  manufacturing sites,  cind  coke-oven
sites.
     Human  exposure to  toluene  ttirough  inhalation  of urban  air is shown  in
Table 10-3.  The concentration of toluene in urban areas in the i'nlted  States in
recent years ranged froa 0.1 tiZ.'a.- to ?'ju  ug/i  (Ts^ie 7-1).  The intake estimate
is based on a  breathing rate cf 1.2 a /hour for an adult  during waking  hours and
O.M a"5 /hour during  sleeping  hours (Siimak,  1930).   It  is  also assumed  that the
sleeping period for an ad'jlt is 8 hours/day.  This  results in an inspire;!  volume
of M.2 x  16 x 7 •» 0.^. x fc x 7)  -  15C.6 n-.-'/week.
     ftear user sites,  the range  of toluene  concentration  has been  assumed to be
5.5 to  600  pg/i".   This ranje  corresponds  to the measured  value  of Sexton and
Westberg  (19t53; near-  &n  automotive painting slant  (.Subsection  7.1.1.)  ^solvent
use constitutes about  99t  of  total  usage).  The concentration  of toluene  at  d
distance  18 kE froci the plan*, measured  S5.5 ^g'm ,  a value 10  time«  hit-h^r than
the  background concentration  (Sexton  and  Westberg,  1980).   Therefore,  even
workers who ctxcmute more tnan  '6 km froa> the plant  are suajeptible  to  inhaling
toluene in the concentration range of 5.5 to  600 ,ig/nr  for the entire  168 hours
in a week. The to'uene concentrations near manufacturing sites range from 0.1 to
T<7 W?/m  .  The estimated toluene exposure ran^e from the manufact uri ng and user
sites shown in Table 10-3 is based on a  concentration range of  0.1 to 600
     In rural ind remote areas, the concentration of toluene has been reported to
be in the range of a trace  to  3.8  ng/m^  (Table  7-D.   These concentrations were
determined in 1971; the current level may be lower than this range,  as  indicated
                                      10-7

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                                                    TABLE 10-3

                               Toluene  Exposure  Under  Different  Exposure  Scenarios
Scenario
General Population
Inhalation
Urban areas
Rural and remote areas
Areas near manufacturing
and user sites
Ingestion
_. Drinking water
9 Food (fish only)
OD
Occupational Group
Inhalation
Dermal
Cigarette Smokers
Inhalation
Observed
Rar?e of
Concentration
0. V to 204 |ig/m3
trace to 3-8 |jg/m
0.1 to 600 ng/m

0 to 19 pg/Jl
0 to 1 mg/kg
377,000 tig/m3
0 to 170 ng/«,
0.1 mg/cigarette
Frequency Total Volume
of Exposed or Inhalation or
Exposure Amount Consumed Ingestion Rate
(tag/wk)
168 h/wk
168 h/wk
168 h/wk

2 i/a
6.5 g/d
40 h/d
0 to 30 min/wk
20 cigarettes/d
156.8 m3 0.02 to 32
156.8 m3 trace 0.6
156.8 m3 0.02 to 9"

14 t 0 to 0.3
45.5 g 0 to 0.45
iJS ra3 18,100
5.9 1 0 to 1.0
110 cigarettes 1U
  This  value  is the OSHA  recommended  standard and represents  the  worst-case  estimate.   Ir some  industries,  the
  exposure level rarely exceeds  10 pj/o>.
  This value represents exposure to blood due  to dermal contact and represent absorbed lerels.
c This value is the OSHA recommended standard.
h = hour; wk = week; d = day; min = minute

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by the toluene concentration  reported at Grand  Canyon  in 1979.  The estimated
tolume  exposure  in rural and remote areas is shown in Table 10-3.
     It  should be remembered that Table 10-3 shows the amount of toluene  inspired
per week by humans around certain exposure scenarios and not  the amount absorbed.
Only a  certain fraction  of  the toluene  inhaled is  absorbed  by human organs.
Also, part of the absorbed toluene is rapidly excreted from the  body.
10.2.  INGESTION  EXPOSURE BASED ON MONITORING DATA
     No  theoretical modeling method  is available  for estimating toluene  exposure
from various  ingestion sources.   Therefore,  the  exposure  estimate  from  this
source has been attempted by using  the  limited monitoring data that are avail-
able.
10.2.1.   Exposure from Drinking Water.  The concentrations of toluene in  drinking
water range  from  0  to 19 ng/S.  (Subsection  7.1.2.5.).   The  concentration  of
toluene measured  in well waters in New York State was below  10 ng/fc  (Subsection
7.1.2.1).).  Therefore, a concentration range of  0  to 19  ng/£  has been  used  for
exposure assessment shown in Table 10-3.   A consumption rate of .? 2./day  has  also
been assumed for  exposure asessment.
 10.2.2.    Exposure  from Edible  Aquatic  Organisms.   The  concentration  range  of
toluene in edible aquatic  organisms has been assumed to be 0 to  1  mg/kg,  based on
the  level of toluene found in unspecified fish tissues (Subsection  T.l.'O.   On
the  basis of  these data  and  the assumption that the  per capita consumption  of
aquatic  organisms  in  the United States is approximately  6.5 g/day  (Stephan,
 1980),  the exposure range of toluene from food is shown  in  Table  10-3.
 10.3.   OCCUPATIONAL EXPOSURE
     Occupational  exposure to  toluene can primarily take place  from  inhalation
 of air  containing  toluene and  from  skin contact with toluene  or other solvent
 mixtures containing toluene.   The concentration of  toluene in the air of the  work
 place has 'been assumed to be  377,000 ng/m .  This value corresponds  to the NIOSH
 (National Institute for Occupational Safety and Health) recommended  workroom air
 standard of 100 ppm toluene vapor as a time-weighted  average (TWA)  exposure for
 an 8-hour work day (NIOSH, 1973).  Thi-?  value  is reasonably  close to  the  actual
 occupational  exposure levels  discuised in  Section 7.2.   Based on  the above
 assumptions, the  inhalation exposure of  toluene  by occupational  groups  as shown
 in Table 10-3 far exceeds that  for any other group.
     Sato and  Nakajima (1978)  studied the absorption of toluene through human
 skin.   These investigators immersed one  hand of  5 male subjects in  pure toluene
                                      10-9

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for 30 minutes  and  monitored the blood levels of toluene.  A peak concentration
of 170 \ig/"i of  blood  was observed  after  a 30-minute  immersion.   This maximum
concentration was maintained for 10 to 15  minutes  after  exposure had ended and
decreased thereafter.
     Although   the  standard  proposed  by  NIOSH  (1973)   requires all  workers
handling toluene to wear gloves, it is conceivable that  short-tern exposure of
bare skin to toluene takes place under certain  circumstances.  For assessment of
exposure through skin as shown in Table  10-3,  a  maximum  concentration of 170 ug/J,
in blood and a  blood volume of 5.9 V. for an adult male have been  assumed.   It has
also  been  assumed that  the  skin  exposure  duration  does  not  exceed  30
minutes/week.     It  also  should  be  recognized  that  the  value  for  blood
concentration  through  dermal contact given in Table 10-3 does not    -resent the
total exposure  value,  as it  ignores exposure to other  organs.
10.1*.  CIGARETTE SMOKERS
     The concentration of toluene in inhaled cigarette  smoke has been determined
to be 0.1 mg/cigarette (see  Subsection 7.3).  In assessing toluene exposure from
cigarette smoking,  it  was assumed  that  an individual  saokes  20 cigarettes per
day.  On the basis of these assumptions, it  can  be predicted from Table 10-3 that
cigarette smoking may  be the second  largest source  of human exposure to toluene.
10.5.  LIMITATIONS  OF  EXPOSURE ESTIMATE BASED ON MONITORING DATA
     As discussed earlier, exposure estimates  on  the  basis of monitoring  data
have the following  listitations:

        (1)  The limited monitoring data do not  provide information
             for   estimating   exposure   under  different   exposure
             scenarios.  Even when  some  data  are  available,  they may
             be inadequate and even susceptible to error.  It is very
            difficult to assess the errors in the monitoring data.

        (2) The monitoring  data often do not relate to the source of
            emissions  in terms of material balancing of the amount
            emitted  and the concentration measured.

        (3) The population  distribution around the monitoring area is
            rarely provided in  these data, although such data may be
            available independently of monitoring.
                                10-10

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       (M)  The estimate  for  toluene exposure to the general popula-
            tion from food and drinking water as given in Table 10-3,
            is very crude,  Tolut,*>e  has been detected in only a small
            fraction of total drinking water supplies monitored (Sub-
            section  7.1.2.5.).    The  exposure  estimate  does  not
    \        specify  either the  number  of people  or the  locations
            where  people  are  exposed to toluene from drinking water.
            The  same can  be  saitf with  respect to  toluene  exposure
            fro»  food.

10.6.   COMPARISON  BETWEEN  EXPOSURE DATA BASED ON THEORETICAL AND EXPERIMENTAL
       VALUES
     If the  concentration  values  ranging from 0 ng/nr' to greater than 100 ug/rc
(Table 10-2) are combined  with the value of 156.8 m  for inspired volume of air
per week,  an  inhalation  exposure  estimate   33  shown  in  Table  10-4  can  be
developed.
                                  TABLE
  Exposed  Population and Exposed Amount of Toluene From Dispersion Modelling
     Concentration
        Level                                              Exposed Concentration
                                                                 mg/week
     MOO                                                    >15.7
      100 to 10                                               15.7 to 1.6
       10 to 1                                                  1.6 to 0.15
        1 to 0.1                                                0.15 to 0.02
        0. 1  to  0                                                0.02 to 0

aSour ce :   S 1 i mak ,  1 9flO
                                     10-11

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     A  comparison of  inhalation exposure  data shown  in  Table 10-^,  which  are
based on dispersion  equations,  with  inhalation  exposure data  in Table  10-3,
which are  derived  from  monitored  concentrations,  shows  reasonable agreement
between  the two sets of data..  The monitoring data  estimate toluene  inhalation by
the general population in  urban areas to  be 0.02 to 32 tig/week.  The  exposure
data developed from dispersion equations estimate this  value to  be in the  range
of zero  to  greater  than  15.7  s^/week.   The  cumulative  inhalation  exposure can be
calculated  by multiplying  the exposed concentrations  from Table TO-M  with  the
appropriate exposed population given in Table  10-2.

10.7. REFERENCES

ANDERSON, G.E.,  LIU,  C.S.,  HOLMAIi,  H.V. and KILLUS, J. P.  (1980).   Human  Exposure
to  Atmospheric Concentrations of Selected  Chemicals,  Publication No.  unavail-
able.  Prepared  by  Systems  Applications.  Inc.,  San Rafael, CA,  under Contract  No.
EPA  68-02-3066.   U.S. Environmental Protection Agency, Research Triangle  Park,
NC.

NIOSH (NATIONAL  INSTITUTE FOR OCCUPATIONAL  SAFETY  AND HEALTH).  (1973).   Criteria
for a Recommended Standard.  Occupational Exposure to Toluene.   HEW Publ. No.  HMS
73-11023 'u.3. Gov't.  Printing Office, Washington, DC.

SLIMAK,  M.  (1980).  Exposure Assessment of Priority  Pollutants:  Toluene.  Draft
report prepared  by Arthur D.  Little, Inc.,  Cambridge, MA,  for the U.S.   Environ-
mental Protection Agency, Monitoring and Data  Support  Division,  Washington,  DC.

SATO, A. ar.d NAKAJIMA,   T.   M978).   Differences follwoing skin or  inhalation
exposure in  the absorption   and  excretion  kinetics   of  trichloroethylene  and
toluene.  Brit.  J.  Ind.  Med.   ^5: U3-^9.

SEXTON,  K.  and WES^BERG,  H.  (1980).  Ambient hydrocarbon  arid  ozone measurements
downwind of a large automotive painting plant.   Environ. Sci.  Technol.   1jl_: 329.
(Cited in Syracuse Research Corporation, 1980).
                                     10-12

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STEPHAN, C.E.  (I960).  Memorandum to J.  Stara,  U.S.  EPA,  July 3.  as cited in
U.S.   EPA,  1980.

TURNER, D.B.  (1969).  Workbook of Atmospheric Dispersion Estimates. U.S.  Dept.
of Health,  Education,  and  Welfare,  Revised,  1969.  (Cited in Walker, 1976).

WALKER,  P.   (1976).   Air  Pollution Assessment  of  Toluene.   Report prepared by
Mitre  Corporation.     Prepared  for  U.S.,  Environmental  Protection  Agency.
Available through  NTIS Order No.   PB 256735,  Springfield, VA.
                                     10-13

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                            11.  EFFECTS ON HUMANS

     Hunan exposure to toluene primarily involves  inhalation, and  consequently
the effect  of  greatest concern  is  dysfunction  of the  CNS.    As  detailed  in
Chapters 9 and  10,  millions of  individuals are exposed to toluene via inhalation
of air frcxi ambient atmosphere and cigarette smoke (ppb concentrations),  and from
occupational exposures (ppm  concentrations).   T-'d.city studies of  humans  have
centered,  however,  on  evaluation  of  individuals  exposed   to   toluene   in
experimental and   occupational  settings,  and  from  deliberate  inhalation  of
toluene or toluene-containing substances  ("glue sniffing").  It  should  be  noted
that occupational  exposures  and glue sniffing often involve  complex mixtures  of
solvents, and  that prior to  the 1950s, benzene  was a  common  contaminant  of
toluene.   In evaluating the  effects  of toluene  exposures,  the purity of  the
compound must be considered.
     Glue sniffers  inhale the vapors from a wide variety of volatile hydrocarbons
(usually  poorly defined mixtures)  contained  in  products  such as  glues  and
thinners for their euphoric  or intoxicating  effects.   The most  popular  of  these
products contains  toluene, and  toluene is the hydrocarbon most frequently impli-
cated as the cause  of  the adverse effects associated with deliberate inhalation.
The practice has  been reviewed extensively  (Massengale,  1963;  Barman  et  al.,
1964; Press  and Done, 196?a,  1967b;  Gellman,   i968;  Wyse,  1973; Linder,  1975;
Faillace andGuynn, 1976; Oliver and Watscn,  1977;  Walter et al.,  1977;  Watson,
1979).   Excessive  levels of  toluene  generally are  inhaled over a  short  time
interval, and repeated inhalation of the  vapors is associated with  the  develop-
ment  of  tolerance  and psychological  dependence.  The  most common  methods  of
inhalation involve (1) placing the  solvent  in  a  plastic  bag and  inhaling  the
funes, (2) soaking a rag or  handkerchief  with the solvent and sniffing  the rag,
and (3) sniffing the  solvent from a container.  The concentrations  of  toluene
inhaled under these conditions  can approach 30,000 ppm (i.e., saturation concen-
tration at 20°C).
11.1.  EFFECTS  ON  THE  NERVOUS  SYSTEM
11.1.1.  Central Nervous System
     11.1.1.1.   ACUTE  EFFECTS  — Experimental exposures of up to 800 ppm toluene
have produced acute dose-related CNS alterations  (Von Oettingen  et  al., 1942a,
1942b; Carpenter et al., 1944).  Von Oettingen et  al.   (1942a,  1942b)  provided
                                     11-1

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what generally is acknowledged  to be the most complete description of the effects
of pure toluene  (benzene £ 0.01$)  on the CNS.   In single 8-hour exposures,  3
human subjects were subjected to Concentrations of toluene in an exposure chamber
that ranged from 50 to 800  ppm  (Table 11-1).  A maximum of two exposures a week
were  conducted  to allow sufficient  time in  between for  recovery;  a total  of
22 exposures was performed over an 8-week period.  Seven of the ?2 exposures were
to pure air, and exposures  to particular levels of toluene were replicated only 1
to  4  times.   The  effects  observed  are summarized  in  Table 11-1.    Subjective
complaints of fatigue, muscular weakness, confusion, impaired coordination, and
enlarged  pupils and  accommodation   disturbances  were  reported at  levels  of
200 ppra.    These  effects  increased  in  severity  with  increases  in  toluene
concentration,  until   at   800  ppm  the  subjects  experienced  severe  fatigue,
pronounced nausea, mental  confusion,  considerable incoordination and  staggering
gait,  strongly impaired pupillary  light reflex,  and  after-effects  (muscular
fatigue, nervousness,  and insomnia)  that lasted for several days.
     -Carpenter  and  coworkers  (1944)  exposed  2 male subjects to known  concen-
trations  of  toluene  (purity  not stated) for  periods of 7  to  8 hours  and noted
slight exhilaration at 200 ppa,- and  lassitude, nausea,  and hilarity at 400 ppm.
Lassitude, hilarity, verbosity, and boisterousness  occurred at 600 ppm (anorexia
and  listlessness  were reported as after-effects.), and  transitory  headaches,
extreme  lassitude,  scotomata  (areas  of  depressed  vision),  verbosity,  slight
nausea, and "inebriation" were found at  800 ppm.   Marked  unsteadiness was also
observed  in the subjects  during exposure  to 800 ppm  toluene.   Steadiness was
determined by a test that involved  holding at arms' lergth a wire in  a hole for
3 minutes; the  percentage of time the wire was actually in contact with the side
of  the hole was  determined, and  compared with the normal value  from  each test
session.
      Short-term  experimental exposures  to  toluene  have also elicited increases
in  reaction  time  and  reductions  in  perceptual  speed  (Ogata  ?t  al.,  1970;-
Gamberale  and  Hultengren,  1972).   Ogata and  coworkers  (1970)  reported that 23
Japanese subjects given single exposures  to 200  ppm toluene  showed a prolonged
eye-to-hand reaction time,  but  .no effect  on  flicker  fusion frequenc:   Exposures
were  for 3 hours, or 3 hours and a  1  hour break period followed by  4  additional
hours  of  exposure.   No changes  in either reaction time or  flicker  value were
observed  at  100 ppa.   It  should be  noted,  however, that no  other  information
regarding the design of tnese experiments was presented.
                                      11-2

-------
                                    TABLE 11-1

                    Effects  of Controlled 8  Hour Exposures  to
                      Pure Toluene  on Three Human Subjects3'
Concentration         No.  of                         Effects
                    Exposures


  Ckppm (control)       7       No complaints  or objective symptoms, except  occasional
                              moderate  tiredness toward the  end  of each  exposure,
                              which  was  attributed  to lack  of physical  exercise,
                              unfavorable illumination, and  monotonous noise  from
                              fans.

 50 ppm                2       Drowsiness  with a  very mild headache in 1  subject.  No
                              aftereffects.

100 ppm                4       Moderate  fatigue  and sleepiness  (3), and a  slight
                              headache  on one occasion (1).

200 ppm                3       Fatigue   (3),  muscular weakness  (2),  confusion  (2),
                              impaired  coordination  (2),  paresthesia  of the  skin
                              (2),  repeated  headache (1),  and nausea (1)  at  the end
    i                          of the exposure.   In several  instances, the  pupils
                              were  dilated,  pupillary  light reflex was impaired, and
                              the fuhdus  of the  eye  was  engorged.    Aftereffects
                              included    fatigue,    general    confusion,   moderate
                              insomnia, and  restless sleep in all 3  subjects.
300 ppm                2       Severe fatigue  (3),  headache  (2), muscular weakness
                              and incoordination (1),  and  slight  pallor of the  eye-
                              ground (2).   Aftereffects  included  fatigue  (3)  and
                              insomnia  (1).

HOO ppm                2       Fatigue   and mental  confusion  .(3), headache,   pares-
                              thesia of the skin, muscular weakness,  dilated  pupils,
                              and pale  eyeground  (2).   Aftereffects  were  fatigue
                              (3),  skin paresthesia (1),  headache (1),  and insomnia
                              (2).
600 ppm                1       Extreme   fatigue,  mental   confusion,  exhilaration,
                              nausea,  headache  and dizziness  (3), and  severe  head-
                              ache   (2)  after 3  hours  of  exposure.  After 8 hours'
                              exposure,  the effects included considerable  incoor-
                              dination    and  staggering   gait   (3),   and   several
                              instances of dilated pupils, impaired  pupillary light
                              reflex   and pale optic  discs;  aftereffects included
                              fatigue   and weakness,  nausea,  nervousness and  some
                              confusion (3), severe headache (2), and insomnia  (2).
                              Fatigue   and nervousness persisted  on the  following
                              day.
                                        11-3

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                                TABLE 11-1 (cpnt.)
Concentration         No.  of                         Effects
                    Exposures


800 ppm                1       Rapid onset  of severe  fatigue and,  after  3 hours,
                              pronounced nausea,  confusion,  lack of  self-control,
                              and considerable incoordination and staggering gait in
                              all 3  subjects.    Also,  pupillary  light  reflex  was
                              strongly impaired  (1) and optic discs were  pale  (2).
                              All 3 subjects showed considerable  aftereffects, last-
                              ing at  least several days, which included  severe  ner-
                              vousness,  muscular fatigue,  and insomnia.


aSource:   Von Oettingen et al.,  1942a, 19M2b
 Exposures were twice weekly  for 8 weeks.   The  number of subjects  affected is
 noted in parentheses.

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     In a more extensive  study,  Qamberale and Hultengren (1972)  exposed  12 maJ e
subjects to 100, 300, 500, or 700 ppm  toluene (via breathing valve and mouth-
piece) during successive  20-minute exposure periods, and measured  their  perfor-
mance on  four  tests of  perceptual  speed  and  reaction  time at  each  level  of
exposure (Table 11-2).   The tests were always made in the  same  sequence  (i.e.,
Identical Numbers,  Spokes, Simple Reaction Time,  Choice Reaction Time) during
the  final  15 minutes  of each  exposure  period.   Toluene concentrations were
increased from TOO  to 300 ppa and from  500 to 700  ppm without interruption,  but
the increase from 300 to  500  ppm was made following a 5-minute interval  without
exposure.  Menthol  crystals contained in the  mouthpiece  tubing  camouflaged  the
taste and smell of  the  toluene.   The 12 subjects were divided into two  groups; of
equal size:  subjects in one group were studied individually, first  under  experi-
mental conditions with  exposure  and  then under control  (atmospheric air contain^-
ing menthol) conditions  7 days  later,  while subjects in  the  other group were
studied under similar conditions but in the reverse order. The camouflage of  the
inspiratory air with menthol  made it impossible for 11  of  the  12 subjects  to
distinguish between exposure  to  toluene and exposure  to  pure air.
     Results of the Gamberale  and Hultengren  (1972) study showed  that both reac-
tion  time and  perceptual  speed were  impaired during  exposure  to toluene  as
compared to exposure to nure air  (Table 11-2).  V.'ith respect to reaction  time,  a
significant effect  was  noted  upon exposure to 300 ppm toluene in one test  (Simple
Reaction Time), and a performance aecrsnent, which  reportedly approached  statis-
tical significance  at the 0.05 level, was noted for the other test  (Choice Reac-
tion Time).   Subject reaction  time  was  further impaired  at  higher  levels  of
exposure  (500 and  700  ppm  toluene),  but  no impairment  in  either  reaction time
test was noted for exposure  to  100  ppm.   (The 100 ppm reaction time  no-effect
level is consistent witn  the  aforementioned results of Ogata et  al., 1970.)   No
statistically significant impairment in  subject  perceptual speed was observed
until the concentration of toluene in the inspiratory air was 700  pptr.   Because
perceptual speed was unaffected at  concentrations below  700  ppm, the  authors
suggested  that  the simpler CNS  functions  may be  affected  at  lower levels  of
toluene exposure than the more complex  functions.
     Winneke et al. (1976) noted, in an abstract published in the Proceedings of
the 2nd International Industrial and Environmental Neurology Congress (Prague,
Czechoslovakia), that  experimental  exposure to 98 ppm  toluene for 3 hours  did
not  affect  psychophysiological   performance  in 20  subjects.    The parameters
                                     11-5

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                                                  TABLE  11-2
                                                                                                   a, ti
          Effect  of Toluene  Exposure  on  the  Performance of  Perceotual Sp«ed  and  Reaction Tloe Testa
Mean Test Scores

Performance Teat

Ider.tical Numbers0
(minutes)


Spolces
(seconds)


Reaction Time - Simple
(neters/second)


Reaction Tim? - Choice
(asters/second)


Source: Gaaberale and
t

Concentration
(ppm)
100
300
500
700
100
300
500
700
100
300
500
700
100
300
500
700
Hultei.gren, 197?

Fsperloental
Conditions

5.62
5.25
5.13
5.19
50.5
16.7
13.6
15-1
228
236
216
253
125
129
132
112

ir*» t nrto .-, f 1AH iri(\ C
Control (Air)
Conditions

5.53
5.29
5.01
1.80
50,8
13-7
10.2
36.9
230
222
219
211
122
116
100
108

inn n r. i4 *T n /"* «. .-.™ A, .—

t-Value

+0.50
-0-39
+ 1.31
+2. 65*
-0.08
+ 1.18
+ 1.28
+2.51"
-0.31
+2.35*
+ 3.88"
+1.8l»»
+0.31*
+ 1.99
+2.91»
+3.«iS««


 minute periods.  The ^sts were pet ~   i^ed at each concentration Sbquentially in the order listed.  The number of
 times sacn test sequence was repeated waa not stated.

 Perceptual speed:   Identical Numbers.  Subjects were  Instructed tc underline the 3-d!git number, fros a total of
 60 columns, that was identical to the number at the head of each column.  Performance was assured as the tioc
 taken to complete the test.

 Perceptual speed:   Spokes.  Sjbjects were Instructed to connect circles located at  rornJoa on four pages and
 numbered froa 1  to 20 In the correct  numerical order using a pen.  Perfi.mnee was measured  as the oean time taken
 for the four assignments.

 Siople Reaction Tine.  Subjects were instructed tc respond to a signal  from a lamp  by pressing a pushbutton.
 Stimuli were administered  at  Intervals of approxlmntely  10  aeron-tj.i,  pn  acoustic  warning signal Mas  given  3
 seconds prior to onset of stimuli, and 30 stiicull were given ]n each trial.  Performance was measured as the meen
 reaction time for the last 20 stlnull administered.

 Choice Reaction Time:   Stimulus/reply test as above, but there were  three pushbuttons »c.ulpp«d with Batching
 stimulus lamps.  Stimulus administration followed a random sequence  with the number of light signals evenly
 distributed among the lamps, but the trial and p-"rforffiance measurements were otherwise the s^jce as for simple
 reaction time.
Degrees of freedom = 11;  »P < 0.05; "f < 0.01; "«P < 0.001

-------
evaluated in t.Ms study included performance in a bisensory (auditory and visual)
vigilance task, psychomotor performance, critica]  flicker  frequency,  and  audi-
tory evoked potentials.   The available abstract did not  provide  any  additional
information on  the  experimental  design,  the r.^ture of  the  psyr.hrphysiological
tests, or the results of tnis  study.
     Gusev (1965)  examined  the effects of acute low-level  toluene exposure on the
electroencephalographic  (EEC)  activity of four human subjects who were trained
to develop synchronous and wel i-marked alpha  rhythms when  stieulated  by light.
Toluene exposures of approximately 0.27 ppm (1 cig/m ) for  6  minutes  apparently
caused statistically distinct changes  in  EEG  activity fron  the  left  tesjporal-
occipital region  in  all  subjects; these  changes  persisted through a  6-minutfr
recovery period.  It should be noted that the 0.27 ppm concentration is slightly
lower than the odor threshold determined  for toluene  in  the sant  experiment
0.40 ppm;  see   subsection 11.7.2.).     Toluene   concentration-;   of   0.16 ppm
(0.6 mg/m ) caused no variations in the electric potentials of the EEC's.  Expo-
sure sessions consisted  of  10 separate observation periods in which inhalation of
toluene (5 periods)  alternated with inhalation of pure air (5 periods).  A single
peri-od consisted of  18 one-minute  cycles.   Every  cycle  included  the  sequential
presentation of a sound  stimulus (10 seconds),  a  wait  for the  light  stimulus
(7 seconds),  the presentation of  the light stimulus (18 seconds), and an interval
of a-.tive physical exercise (25 seconds)  for recovery  of normal EEG rhythm.   Of
the IB minutes allotted  for EEG recording in each period,  3 minutes were used for
training,  the  next  3 minutes  for   background  observations,   the   following
6 minutes  for   the  toluene exposure,  and  the  final   6 minutes   for  recovery.
Although the reported effects  on EEG  activity may represent a subtle  indication
cf perception,  there is  no apparent toxicological  significance to the finding.
It should further be noted that western studies  have not reported any effect of
toluene on the  CNS  at  such low levels of  exposure, and  that  the  purity of  the
toluene used was not stated.
     Narcosis is the primary result of  acute toluene exposure at high concentra-
tions.  A number of accounts of workers who were  rendered unconscious  by toluene
vapor have been published in the medical literature (Lurie,  19^9; Browning, 1965;
Longley et al., 1967; Reisin et al.,  1975).  Host  of these  cases  have involved
the entry  of  workmen into confined areas with  poor ventilation  and  subsequent
exposure to high levels of toluene during maintenance operations.  Longley et al.
(1967) described two episodes  of  acute  toluene intoxication involving 26 men who
                                      11-7

-------
were exposed in the  holds of cargo ships.  Toluene concentrations  were  estimated
to have ranged from 10,000  ppm at waist level  to 30,000 ppm at floor level,  but
it was emphasized that this estimate was  purely  conjectural.   Effects  at  these
concentrations  ranged  from exhilaration,  lightheadedness,  and  clumsiness  and
dizziness to collapse and unconsciousness.  No deaths occurred and recovery  was
quite  rapid,  with  no after-effects  following  removal  from  the  contaminated
atmosphere.   The curations of the  exposures  were not  indicated,  but loss  of
consciousness occurred within minutes.
     Episodes of toluene abuse are characterised by the progressive development
of CNS symptoms*  Toluene sniffers axperience an initial excitatory stage that is
typically characterized  by drunkenness,  dizziness,  euphoria,  delusions, nausea
and vooiting, and, less commonly, visual and auditory hallucinations (Press  and
Done,  19&7a,  I967b;  Wyse, 1973; Lewis and Patterson, 1971*;  Hayden et ai.,  1977;
Oliver and  Watson,  1977; Barnes,  1979).   As  duration of exposure increases,
symptoms indicative of CNS  depression  .become evident:  confusion and disorienta-
tion, headache, blurred vision and reduced speech, drowsiness, muscular incoor-
dination, ataxia, depressed reflexes, and nystagmus.  In extreme  cases, loss of
consciousness, possibly with  convulsions  (Helliwell  and Murphy,  1979), occurs.
The  duration  and severity of these  effects  vary greatly, depending  upon  the
intensity of  exposure; the duration may range frctn  15 minutes to a few  hours
(Press and  Done, 1967b).  Also,  all of  the  symptoms described  have  not  been
exhibited in any single sniffer, nor in any single episode of sniffing.
     Winek et ai . ( 1968)  published partial results of an autopsy on an adolescent
who had died as a result  of sniffing model airplane glue containing toluene.   At
autopsy,  the  cut surfaces of the lungs  of this  individual  were found  to be
extremely frothy and congested, with diminished amounts of crepitation  through-
out  the  lung tissue.  Other  gross observations that  were noted included some
petechial hemorrhages in  the larynx and upper trachea, firmness and congestion in
the spleen, and a dark, red-brown color and congestion in the liver.  No hemor-
rhages, obstructions, or ulcerations were seer, anywhere in the gastrointestinal
tract, and  all other organs  were unremarkable.   The  results  of toxicological
analyses  of  various body tissues  for toluene are  presented in Section  13.2.
Congestion  in various  organs, swelling of  the  brain,  subseromucous petechiae,
and  pulmonary edema v:ere associated  With 19  other  cases  of acute death from
thinner intoxication (Chiba,   1969); the English abstract of this  Japanese study
indicated that toluene was  the major  component of the inhaled thinner.   Nomiyama
                                      11-8

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and Nomiyama  (1978) described an instance in which 1 adolescents were  found dead
after sniffing 99J pure toluene in  a  car,  but post-mortem results  utrier than
levels of toluene (blood and alveolar  air)  and hippuric acid (urine) were not
presented.  Sudden death due to solvent sniffing has  been  reported in at least
122  cases  (Bass,  1970;  Alha  et  al.,  1973).    The  sudden  deaths  have  been
attriouted, however,  to severe  cardiac arrhythmia,  and are discussed in  Sub-
section 11,5. (Effects  on  the Heart).
     11.1.1.2.   SUBCHRONIC AND  CHRONIC EFFECTS —  Wilson  (19^3) described the
effects of exposure to commercial toluene vapor  on 100 workers (out of  a total of
100C workers) who showed symptoms  severe enough to  seek examination at a hospi-
tal.  The workers were exposed daily to  toluene concentrations ranging  from 50 to
1500 ppn  for  periods  of  1  to  3 weeks, but  the  composition of  the  commercial
formulation and the type of industry were not  described.   Also, it  is unclear
wnether the  remaining  900  workers  evidenced any symptoms  of toluene exposure.
The  concentration  of  toluene was  determined shortly  after any  dxnosed person
appeared at  the  hospital  with  symptoms, and the patients  were  classified into
groups by degree of exposure.   The following effects were  reported:

     50 to 200  ppm (approximately 60J of the  patients) - headache, lassitude, and
loss of appetite.  These symptoms  were so mild that they were considered to be
due  primarily to psychogenic and other  factors  rather  than  to toluene fumes.

     200 to 500 ppm (approximately 30J  of  the patients) -  headache, nausea, bad
taste in the  mouth, anorexia, lassitude, slight but definite impairment of coor-
dination and  reaction time, and momentary  loss  of memory.

     500 to  1500  ppm  (approximately  10? of  the  patients)  - nausea,  headache,
dizziness, anorexia,  palpitation,  and  extreme  weakness.   Loss  of coordination
was  pronounced and reaction time was  definitely impaired.

     Characteristic CNS alterations  have also been  described in  foreign reports
of  workers  exposed  for   longer  durations  to  moderate  levels of toluene.
Parmeggiani and Sassi  (1954) found signs of "nervous hyperexcitability" in 6 out
of  11 paint and pharmaceutical  industry workers who  were exposed  to 200-800 ppm
toluene vapor for "many" years. Capellini  and  Alessio (197D noted symptoms of
stupor, nervousness,  and insomnia  in one worker who was employed for "diverse"
                                     11-9

-------
years in preparing a toluene-containing  mixture for use in the manufacture  of
V-belts.  The mean  atnospheric concentration of  toluene in the mixing department
was 250 ppai,  with extremes of 210 ppm and  300 ppcn.  No CNS effects were observed,
however, in  1? other workers who were  exposed  to 125  ppm toluene  (range,  60  to
160 ppm) while engaged  in the manufacture of  the belts.
     In a more extensive study,  Suhr (1975)  found no evidence  of adverse neuro-
logical effects in  a  group of 100 rotogravure printers with at  least  10 years  of
exposure to 200 to 400  ppai pure  toluene (<0.3J benzene).  Subjective complaints
indicative of CNS toxicity  (headache,  giddiness,  nervousness,  irritability,
sleeplessness, bodily  fatigue  and  incoordination),  abnormal  reflex reactions,
and abnormal  Sphallograph test results were not  found to occur significantly more
often  in  the printers  t:^an  in  an  unexposed  control group of  equal  size.  The
Sphallograph  is an instrument  that  is  used  to  detect  slight disturbances  of
muscular coordination by sensing variation in the balance of two metal plates;  a
test person stands  on the plates, and balance disturbances are detected by strain
gauges.
     The Suhr  (1975)  conclusion that  chronic  occupational  exposure to 200  to
iiOO ppm toluene did  not  cause  adverse neurological  effects in the  rotogravure
workers is equivocal  for several  reasons.  First, the nature of the  control  group
used in this  study  is not defined, other than that they "were from  the same firm
and not exposed to.toluene."  Additionally, the worker  and  control  groups were
only roughly  matched  by groups  for age   distribution,  years  of exposure, and
nature of workshift (i.e.,  2 or  3 shift work).  Second,  the  venous  blood levels
measured in the printing room workers at  tne  end of  their shifts indicate  expo-
sure to toluene levels of at least 300 ppna and possibly   as   high   as  600 ppm.
These  levels  are  consistent  with the reported  air  concentration  measurements,
which  were made with a "measuring cell"   device.  It  is  unclear,  however,  when
workers were  examined for reflex reactions and Sphallograph measurements.  If  it
was after or  before the  workshifts  (as the data for the 33  Sphallograph groups
would  indicate), then  blood levels  of  toluene  may have  declined significantly.
Astrand et al. (1972) have shown major drops  in levels within  minutes after the
removal of human subjects from exposure.  Third,  the  Sphallograph appears to be a
very infrequently  used  device in the United States;  several behavioral toxicolo-
gists who were contacted by Syracuse Research  Corporation  (SRC) indicated that
they have never heard of the instrument,  and  the device  does  not appear to have
been described in  standard texts.  Suhr (1975)  also cites the work  of Pohl and
                                     11-10

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Schnilde  (1973)i  who tested the effects of "extreme"  concentrations  of  11  fre-
quently used organic  solvents  in humans  with the Sphallograph and  fc fid  only
minimal effects.  This would argue that the Sphallograph is not a sensitive  test
for determining CNS effects of solvents.  Last,  until  more is known concerning
the exposures of  the control  group,  the significance of the reportedly negative
results of the  .subjective symptom survey is  questionable.
     Chronic occupational  exposure  to  toluene  has also  been  associated  with
behavioral changes.  Munchinger (1963) diagnosed an "organic paychosyndrome"  in
21$ of a group of printers exposed  on the average  to 300 ppra toluene for 18 years
(mean  age,  42  years),  and in  40% of  a group of printers' helpers  exposed  to
430 ppm for 12  years (mean age,  414 years).  A total  of 110 workers were examined,
but the number of  printers and printers'  helpers was  not. stated and testing on
control subjects  was not performed. The syndrome  was characterized by subjective
memory, thinking, and activity disturbances.   Results of Rorschach testing  were
consistent with the psychosyndrome diagnosis  in  83? of the cases.   In combina-
tion, Rorschach Test and Knoepfel's 13-Error Test results  agreed with the diagno-
sis in 95J of the cases.
     More  recently, several  groups  of investigators have  shown that long-term
exposure to  combinations  of  toluene  and  other common organic  solvents  caused
impairments in visual intelligence and  psychomotor  performance of  workers.   In
1973, Lindstroeia  compared the psychological  test performances  of a group of 168
male  workers who  had  been exposed to  hydrocarbon  solvents for 0.1  to 30 years
(mean, 6 years) to those  of an  unexposed control group (N - 50).  Twenty-six of
the workers had tx?en exposed  primarily to  toluene and 25 others to a combination
of  toluene and  xylene;  the  remaining workers  (numbers  in  parentheses)  were
exposed primarily to trichloroethylene (4U) , tetrachloroethylene (8), "thinners"
(44),  and miscellaneous  solvents  (21).    Exposure  concentrations  were   not
reported.   Results showed that  the  solvent-exposed  workers  were  inferior  in
performance to the controls in sensorimotor speed performance, psychonotor  per-
formance,  and  visual  accuracy  as determined by standardized  test  procedures
(e.g., Bourdon-Wiersma vigilance  test,  Santa  Ana dexterity test, Mira  psycho-
motor  test).  The  performance of  the  workers  on the Rorsctiach personality  test
was comparable to that of the control  group.
     Hanninen et  al. (1976) compared the behavioral responses  of a group of 100
car painters with  those  of 101  age-matched nonexposed subjects.   The painters
(mean  age  35  +  11 years) were exposed to different organic  solvents for  1  to
                                     11-11

-------
40 years  (mean,  14.8 JK 8.5 years), but, as detailed in Table  11-3,  toluene was
present in the greatest  amount  (30.6 ppm).

                                 TABLE 11-3
                   Mean  Concentrations  of Organic Solvents
                 in the  Breathing Zone  of  40  Car  Painters3'
                                            Mean
        Solvent                         Concentration
                                           (ppm)

        Toluene                             30.6
        Xylene                               5.8
        Butyl Acetate                        6.8
    i    White Spirit                         4.9
        Methyl Isobutyl Ketone               1.7
        Isopropanol                          .2.9
        Ethyl Acetate                        2,6
        Acetone                              3.1
        Ethanol                              2.9

        g
         Source:   Hanninen et al.,  1976
         Sampling Period = 1  hour;  Number of Car Repair Garages =  6;
         Nunber of Samples =  54.

A battery of  tests  included  1 test for  verbal  intelligence, 3 visual  tests,  5
memory or learning tasks,  4 tests of psychotnotor performances, and  the Rorschach
test for measuring personality changes (Tables 11-4 and 11-5).  Results of  this
study showed significant differences between the exposed and reference  group  ir,
almost all intellectual performances arid memory tasks.  Impairments in visual ;?r: j
verbal intelligence and in memory, as well as  a reduction of emotional reactivity
as  indicated  by  the Rorschach  test,  were  the  predominant effects of  solvent
exposure  (Tables  11-4   and  11-5).     Differences  in  psychotaotor   performances
between the exposed and control  subjects were less consistent;  impairments  were
seen only in some  of the Santa Ana dexterity and finger tapping  test scores, and
                                     11-12

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                                                                 TABLE  11-H
               Perfornance Testa:  Meana, Star.dard Deviations, and Significances Between the Croup Means  (Age-Matched)  Croups*
 I
LO
' Means and Standard Deviations
Test
WAIS8 Similarities test0
WAIS Picture Conpletiond
WAIS Block Design"
Figure Identification
WAIS and WHS8 Digit Spanh
WHS Logical Matory1
HMS Aaaoclate Learning^
Benton Test for Visual Reproduction
Benton Teat for Vl.iual Retention
SADT - right handk
SADT - left nandk
SADT - coordination with both hands'1
Finger Tapping - right hand
Finger Tapping - left hand
Reaction Time (Simple) - right hand
Reaction Time (Simple) - left hand
Reaction Tioe (Choice)
Mlra Teat0
Mira Test"
Exposed (N = 100)
19.1 *_
11.9 »
31. 6 *
32.0 +
10.6 £
11.7 i
15.3 t
21.1 +
8.2 +
11.7 +
12.3 *
29-0 +
202.5 i
186.7 *
12.1 +
12.1 +
9.1 +
18.8 +
2.2 «
3.1
2.9
7.0
9.0
1.6
3-7
3-6
3-1
1.5
5.7
5.1
5-1
29.2
28.5
2.9
3.0
1.8
3.8
1.0
Nonexpoaed (N = 150)
2.9 *
16.2 _*
39.6 »
36.7 *
11.5 +
13.9 «
•7.1 +
22.6 »
8.7 *
17.5 +
U3.6 +
31.5 +
Z'09.6 +
196.1 »
11.9 «
11.7 +
9.1 *;
.?0 . 3 +
2.0 +
2.1
2.3
5.6
9-8
1.8
3-1
2.6
2.3
1.3
5-8
5.1
5.7
23-8
22.1
1.1
1.1
1.2
1.6
0.8
Significance
of Differences
(t-teat)
• «•
1.1
in
• ••
in
ill
• ••
• •i
•
• •

• n

l



«n
i
                Source:  Hanninen et al.,  1976
               Tiechsler Adult Intelligence Scale.
                Measures verbal intelligence and abstraction.
               Veasurea visual Intelligence and observation.
               '"'Measures visual intelligence and abstraction.
                Measures speed of perception and memory for visual  details.
               ^Wechsler Memory Scale.
                Measures memory for digits.
 Measures verbal  memory.
'rlfasurea verbal  memory  and  learning.
 Santa Ana Dexterity Test; measures  psychomotor  speed.
 Measures motor speed.
 Te.3t for nsychomotor behavior  and  psychomotor ability;" two variables  tested.
"paired t-teat.
•P c 0.05; «P <  0.01;  ««P  < 0.001

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                                                  TABLE  11-5

                     Rorschach  Personality Test Variables:  Means, Standard Deviations, and
                           Significances Between  the Group Means— (-Age-Matched  Groups)
J Oi ,._... Significance
Means and Standard Deviations „,. n
-------
reaction times were unaffected by exposure.   It  should be noted that in  other
studies, reaction  time increased  as a  result  of acute  (Ogata  et al.,  1970;
Gamberale  and Hultengren,  1972)  and subchronic  (Wilson,  1973)  exposures  to
toluene  concentrations in excess  of  200 ppm.    The  possible  influence  of
differences  in  initial  intelligence  levels  on the  performance  scores  was
controlled in the Hanninen et al.  (1976) study by a separate comparison of the
test results of 33 pairs of exposed and unexposed subjects who were matched for
age and intelligence.
     In a  related study,  Seppalainen  et  al.  (1978)  examined  the  same  cohort of
car  painters  studied  by Hanninen ar;d  coworkers  (1976) for  neurophysiological
effects.   Results  of  EEC  analysis on  102  sol vent-exposed  car  painters  and 102
nonexposed control subjects showed no increase in abnormalities  (Abnormal  EEGs
were encountered in 32 painters and 37 controls.).  It was noted,  however,  that
the-incidence of abnormal  EEGs  in  both groups was higher than  expected  (approxi-
mately  10?) on the basis of EEG literature.  It was  further reported that  26 of
the  car  painters had  a complex of four  subjective  symptoms indicative of  CNS
disturbance  (interrupted   sleep,  absentmindedness,   easy  to  fall  asleep  when
watching television, frequent headaches);  this symptom complex was found  only in
12 controls.  EEG testing  on the workers with these symptoms showed abnormalities
in 46J (12/26) of the cases, but 26%  (20/76)  of those without the symptom  complex
also  displayed  EEG  abnormalities.    This  difference was  not  statistically
significant  (Chi squared = 2.68).
     Rouskova  (1975) did observe  changes in EEG response to  photic stimulation in
a group of 20 workers  with  a  13-5  year  (average)  history  of  exposure  to higher
concentrations  of  toluene  (>250 ppm)  and 1, 1, 1-trichloroethane  (concentration
not  stated).   Photic  stimulation  was applied in a series  of  rhythmic  flashes,
each lasting  10 seconds with intervals of 10 seconds between each flash  series;
frequencies ranged from 1 to 30 per second.  Evaluated a?  a normal response was
the  occurrence of EEG  activity  of  the same  frequency  as  stimulation or  of  a
harmonic or a subharmonic multiple of that frequency lasting at least one  second.
Results showed that  abnormal  EEG responses were found  in  18  of  the 20  workers
(90$),  but in  only  1 of 20 unexposed control subjects.
     Residual  effects  indicative of  cerebellar  and  cerebral  dysfunction  have
been observed  in a number of persons who had abused toluene or solvent mixtures
containing toluene  over a  period  of years  (Grabski,  1961; Satran  and  Dodson,
1963; Knox and Nelson,  1966; Kelly,  1975;  Boor  and  Hurtig, 1977;  Weisenberger,
                                     11-15

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1977; Keane, 1978; Sasa et al.,  1978;  Tarsh,  1979;  Malm and Lying-Tunell,  1980;
Metrick and Brenner,  1982).  Boor and Hyrtig (1977)  described a case  of  cerebral
involvement in  an  optician  who  regularly used toluene occupationally  to  clean
eyeglasses and contact lenses in a small,  unventilated room, and optic neuropathy
has been observed (Xeane,  1978; Mai in and Lying-Tunell,  I960; Metrick and  Brenner,
1982).   Clinical signs in these individuals included ataxia, intention  tremors,
nystagmus, equilibrium disorders,  positive Babinski reflex, impairment of speech
and hearing, reduced vision, disturbance  of  concentration and memory, emotional
lability,  and  psychosis.   These  reports, which  are summarized in  Table  11-6,
indicate that the severity of the encephalopathic effects generally  varied with
the  intensity  and  duration  of   exposure  and that  the  effects  were largely'
reversible, particularly  when  the exposures were  not  too extreme.   Prolonged
toluene, abuse has, however,  on-occasion led to permanent encephalopathy and  brain
atrophy  as  evidenced  by  EEG   and   neuroradiological   (pneumoencephalogram,
angiograia) changes (Knox  and Nelson,  1966; Boor  and Hurtig,  1977; Sasa et al.,
1978).  Pontomedullary atrophy and abnormal brainsteni auditory evoked potentials
were  recently observed in  two  chronic  toluene  abusers  (Metrick and  Brenner,
1982).
11.1.2.  Peripheral Nervous System.  Matsushita et  al. (1975) found  evidence  of
peripheral  neuropathy in a group of  38 female  shoemakers  (mean  age 20.7 i
5.2 years) who  had been exposed  to a glue containing mainly toluene and  "slight"
gasoline for an average Duration  of 3 years and b months.  The results of neuro-
logical and muscular function tests reportedly showed abnormal tendon reflexes,
reduced grasping power of the dominant hand,  and  decreased finger  tapping  tempo
in  the exposed workers  relative  to  a  group of   16  unexposed control   women
(Table  11-7),  but  descriptions of  the  tests  were not provided.  A  significant
decrease in finger agility was also noted in the  exposed shoemakers;  agility  of
the fingers was estimated by measuring the time needed to move 25  "bulbs"  using
glass chopsticks.   The average toluene concentration in the air varied with time
of  year from  60 to 100 ppm (range 15  to 200 ppm);  in a "few" working places,
gasoline  ranged from 20  to  50 ppm.   An increased urinary  hippuric  acid  level
among the exposed women  (3.26 + 0.8Z mg/ml versus 0.35 + 0.24 mg/ml for controls)
is consistent with an exposure to toluene.
     Electroneuromyographic measurements were made in  the Seppalainen et  al.
(1978)  study  (described in  Section  11.1.1.)  on 59 of the  toluene-exposed car--
painters and 53 referents  with a similar  age distribution for any indication of a
                                     11-16

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                                                                     TABLE  11-6

                                                  Encephalopathic Effects of Chronic Toluene Abuse
Subject (Age)
        Exposure History
          Effects and Diagnosis
                                                                                                                                Reference
Male (33 years)
Hale (30 years)
Fenale (19 years)
Male (25 years)
Pegularly sniffed toluene for I1) years.
Subject purchased a gallon of pure
toluene every 1-6 weeks, and Inhaled the
toluene on an almost dally basis at fre-
quent intervals throughout the day.
10-year history of toluene abuse.
Almost dally sessions of prolonged paint
sniffing for 1-1/2 years.  Ingredients
not specified but It was indicated that
toluene was a common ingredient In all
the brands sniffed.   Previous 1-year
history of multiple drug and solvent
abuse.
10-year history of lacquer thinner (99J
toluene) abuse; during the last 5 years he
had spent virtually all his making hours
Inhaling the vapors (1 gallon used every
2 weeks)
Patient Initially examined after 6 years by
Grsbskl; signs Included ataxla, Intention
tremors, pyramidal signs and psychosis which
were concluded to be consistent with cerebellar
degeneration.  After 8 acre years of abuse, Knox
and Nelson reexamlned the patient and concluded
that the syndrome was primarily a diffuse
cerebral disorder based on findings of ataxla,
treaors, Hob incoordinatlon,  emotional lability,
marked snout reflex, and positive Babinski toe
reflex; cerebral atrophy waa confirmed by EEG
and pneumoencephalography.

Recurrent headaches, "inappropriate" speech,
brief episodes of menory loss,  increased
Irritability, and exaggerated  swings in mood.
Unremarkable clinical and neurological exam,
but nonspecific EEG changes were found that
were regarded as consistent with diffuse
encephalcpathy.

Ataxia, intention tremors of hands and feet,
incoordination, hallucinations,  Nonsal EEG,
brain scan, arterlography, and  pneumoencephalo-
graphy.  The diagnostic impression was
cerebellar dysfunction secondary to scoe toxic
factor in the paint.  Objective neurological
Improvement 5 months after sniffing was
discontinued.

Abasia, mildly slurred speech,  r.ystagmus ,  and
bllatsral  Babinski. signs.   Normal  EEG, nuclide
brain scan, electromyogram, and nerve conduction
studies, but a computerized brain scan-showed
diffuse widening of the cortical and cerebellar
suJci.  Subjective improvement  in condition
following abstinence from exposure, but a
neurolugical exam after 9 months was
ensentially unchanged.
Grabskl, 1961;
Knox and Nelson, 1966
Satran and Dodson, 1963
Kelly, 1975
Boor and Hurtig, 1977

-------
                                                                     Table 11-6.   (cent.)
   Subject  (Age)
        Exposure History
                                                        Effects and Diagnosis
                                                                                                                                  Reference
   Male  (59 years)
   Hals  (age not stated)
   Hale  (27 years)
•J* Hale  (20 years)
cx>
   Hale  (25  years)
    Female (18  years)
Optician who freo.tiently but Inter-
oittep.My used 99i toluene In a small
ur.ventilated roor to clean eytgl-asnes
end contact lenaes.  Unable to smell
toluene because of chror.lc anosnia.
Duration of exposure not stated.

Habitual Inhalation of paint thinner
(toluene) on tha job.  Duration not
stated.

Sniffed unspecified glujs and paint
thinners for 10 years.  From age 25,
toluene was involved ^-5 times per week
(200-300 ml/week used), and from age 26,
he inhaled 1-7 times per day (100 mi/day
used.

3-year history of dally aerosol spray
paint inhalation.  Product contained
copper, toluene, and xylene as solvent.-)
and isobutane propane and roethylene
chloride as propellants.

Sniffed toluene for 1 months, starting
while on the Job using toluene as a
solvent in the rubber processing
industry.
Inhaled pure toluene since age 12,
regularly since age 16 (2 liters us»?d
per month).  Sniffed more heavily th?.ii
usual during the last 2 months.
Fatigue and clumsiness of the left side which
got progressively worse.  Oocaiional staggering
and mildly slurred speech, disturbed concen-
tration and memory.  Normal neurological exam,
EEC, and brain scans.  Dally improvement without
specific treatment following cessation of exposure.

Slzzare behavior prior to hospital admission.
Admitted In an agitated, violent, nearly catatonic
state.

Ann and neck trenors, ataxla, incoordination,
and equilibrium disorders.  Wo abnormal
psychiatric symptoms.  Pneuooencephalographlc
and anglographical evidence of mi dbraln and
cerebrum atrophy.  Degeneration of the
cerebellum suspec'.ed.

Reduced vision, poor color perception, con-
stricted visual fields, normal optic fundl, im-
paired papillary response, ataxia, and nystagmus.
Symptoms slowly subsided following cessation
of paint sniffing.

Delusions and unpredictable behavior.
Largactil prescribed because he was thought to
have a schizophrenic Illness.  Symptoms dis-
appeared and did not recur following termina-
tion of anlffing.

Personality changes (apathy, Irritability,
emotional lability, carelessness), vooitlng,
jlfflculty in walking, and slurred speech
1-2 weeks before admission.  Gait ataxia,
Incoordination, dysarthrla, downbeat nystagmus,
bilateral positive Bablnskl sign, visual and
colu" sense loss, impaired concentration and
abstracting ability upon admission.  Symptoms
consistent with mainly cerebellar-braln stem
involvement and possibly optic neuritis.
Symptoms decreased when she did not Inhale
toluene, and disappeared after 8 months.
Boor and Hurtig,  1977
Welsenberger,  1977
Sasa et al.,  1978
Keane, 1978
Tarsh, 1979
Main and LyIng-Tunell, 1980

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                                   TABLE 11-7

              Results  of Neurological and Muscular Function Tests
                     of Toluene-Exposed  Female Shoemakers
Testb
Abnormal tendon reflex:
. Biceps and triceps
Patellar
Ankle
Pathological reflex
Grasping power (dominant hand)
i
Tapping tempo (M + S.D.)°
I
Cold pressure test
Postural hypotension
Cuff test (upper arm)
Dermatographism
Blocking test (M + S.D.) (seconds)
Numbers investigated
Exposed Group
6(16)
14(37)*
7(18)"
U 3)
11(29)**
162.9 ± 16.6
6(16)
2( 5)
5(13)
5(13)
68.2 + 13-3
38(100)
Cor. ^rol Group
3(19)
K 6)
0( 0)
0( 0)
K 6)
168.6 + 17.3
2(13)
K 6)
K 6)
K 6)
61.8 + 13-7
16(100)
 Source:   Matsushita et al.,  1975
 Numbers  of subjects with abnormal  scores reported.   The percentage  of
 subjects affected is indicated in  the parentheses.
HJnit of  measurement not stated.
Statistical significance (Chi Square- and t-tests):   *P < 0.05;  **P  < 0.1;
 M = mean;  SD =  standard deviation.
                                       11-19

-------
possible peripheral neurotoxic effect from  exposure.   Maximum motor conduction
velocity (MCV), conduction velocity of  the  slower  motor fibers (CVSF),  maximal
sensory conduction velocity (SCV),  and motor distal latencies were recorded from
nerves in the  upper and lower extremities  (median,  ulnar,  deep peroneal,  pos-
terior tibial,  and sural nerves).   Results of these measurements showed that the
mean conduction velocities and motor distal latencies  of the car painters were
almost identical to those recorded for the unexposed control group.  In several
instances,  however, individual nerve  conduction  velocities  were found  to  be
slower than the normal  historical  value  (not stated) for Seppalainen's labora-
tory.  When the conduction velocities of the study group were compared with the
historical  Values,  abnormally - slow MCVs or SCVs and/or  prolonged motor distal
latencies were found in  12 of the  59 painters, but in none of the 53 controls.
     Although  the  two  previous reports  (Matsushita  et al.,  1975;  Seppalainen
et al.,  1978)  indicate  a possible effect of  toluene on the peripheral  nervous
system, toluene's role in the causation  of human peripheral neuropathies has not
been clarified.  Reports of polyneuropathies in abusers  exposed to excessive and
prolonged concentrations of glues  and .solvents  have appeared in the Japanese and
American literature, but have in all cases involved mixtures  of toluene and other
solvents  (Matsumura et  al.,   1972; Takenaka et al.,  1972;  Goto et  al.,  1974;
Shirabe  et  al.,  1974;  Suzuki et al ., 1974;  Korobkin et  al.,  1975;  Oh  and Kim,
1976; Tov»fighi et al.,  19V6; Altenkirch  et  al ,,  1977).   The cases described  in
these reports  were  characterized by the sudden onset and rapid progression of a
symmetric,  predominantly motor polyneuropathy  (although  sensory nerve  involve-
ment of  the glove and stocking type has been reported), even after exposure has
ceased.   Symptoms  included  extremity  weakness, numbness,  paresthesia,  marked
amyotrophy,  and occasional  flaccid  paresis.   Collective results  of  electro-
myographic  studies  have shown  delayed nerve conduction velocities and  signs  of
denervation, and biopsies of nerves have  revealed axonal degeneration, demyelin-
ation, and  enlargement  of some axons with focal accumulation of neurofilaments.
Muscle biopsies revealed extensive neurogenic atrophy.
     The earlier reports regarded  either_n-hexane alone (Korobkin et al., 1975;
Towfighi  et al.,  1976) or  a combination  of  _n-hexane and  toluene (Matsumura
et al.,  1972;  Goto  et al., 1974; Shirabe et al.,  1974;  Suzuki et al.,  1974)  as
the  cause  of  glue sniffers'  neuropathy.  The  following observations have been
offered  as  evidence to  indicate that _n-hexane  plays an important  role  in its
etiology:   (i; in many  of  the reported  cases, neuropathy did not develop until
                                     11-20

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the patients began to sniff glue products that contained jn-hexane,  and (2)  it  is
known that  continuous occupational  exposure  to  n-hexane under  poor  ventilation
conditions produces a neuropathy among workers  that  is  clinically and  patholo-
gically similar to that  observed among the glue sniffers. From a recent outbreak
of  polyneuropathy among  18  glue thinner  sniffers  in  West Germany,  however,
Altenkirch et al. (1977) presented data that  implicate methyl ethyl ketone  (MEK)
as  the  causative  agent  and  argue against n-hexane  and  toluene as the  causes.
These data are summarized as follows (Altenkirch et al., 1977):

          1.   In a number of sniffing adolescents (1000 to 2000), no
               adverse  neurological  effects  were  observed  during  the
               abuse of a thinner  with a  high  _n~hexane   (31$)  and
               toluene  (30$) content over a period of 7 years.

          2.   The   clinical  picture of  neuropathy occurred when  the
               ri-hexane fraction had been  decreased by  approximately
               one-half (16$) and MEK (11$) had  been added;  the  amount
               of toluene was noc significantly changed (29$).

          3.   Individuals who had discontinued sniffing prior  to  the
               introduction  of  the   new  formulation  or who  had used
               only  the old  composition were  not affected.   Neuro-
               pathies  occurred, however,  after  3  to 4 months  in
               sniffers who had used only the new mixture.

          4.   Sniffing even a relatively small  amount of  the MEK-
               containing composition led to neurotoxic damages, while
               comparatively large  amounts  of the  old composition were
               tolerated for a long  time without consequences.

          5.   After  the MEK-containing  thinner  was  taken off  the
               market,  new cases of  the disease were not observed,

Altenkirch and coworkers  (1977) further noted that the exact composition of  the
glues that  contained _n-hexane  and  toluene  cited in  many of the  aforementioned
reports  is  incompletely characterized, and  concluded  that it remains  open  to
                                     11-21

-------
question whether _n-hexane was the sole causative agent in those cases.  It should
be emphasized that no report in which  peripheral neuropathy is attributed, to the
inhalation of toluene alone was located in the literature.   Further, no  sensory
or neuromuscular involvement was  detected  in a patient who experienced permanent
cerebral dysfunction  following prolonged  inhalation  of 99? pure toluene  (Boor
and Hurtig, 1977).
11.2,  EFFECTS ON THE.BLOOD AND HEMATOPOIET1C TISSUE
11.2.1.  Bone Marrow.  The action of  toluene on human  bone  marrow  has been the
subject  of persistent  controversy.   Early  reports  of occupational  exposures
(generally prior to the 1950s) ascribed myelotoxic effects  to toluene (Ferguson
et al.,  1933; Greenburg et al., 19^2; Wilson, 19^3),  but the majority of recent
evidence .Indicates that the chemical  is not toxic to the blood  or  bone  marrow.
The myelotoxic  effects  previously attributed  to  toluene  are generally regarded
by recent investigators to  have  been  the  result  of concurrent exposure  to  ben-
zene,  which  was present as  a  contaminant.   Banfer (1961)  noted that it  first
became  possible  to supply industry with adequate quantities of "pure"  toluene
C<0.3J benzene) in 1955; earlier, workers  were typically exposed to toluene that
was derived from coal tar and contaminated with as much as  20?  benzene.
     Greenburg et al. (19^2) found mild depression of erythrocyte levels,  abso-
lute lymphocytos^s, macrooytosis, and elevation of the  hemoglobin level  and the
mean corpuscular hemoglobin  concentration in  61  airplane painters  who had  been
exposed to 100 to 1100 ppm toluene for periods extending from 2 weeks to  5 years
(Table  11-8)-  Exposure was also associated with liver  enlargement  in 13 of the
61  painters   (Section  11.3.), but  not  with abnormal  granulocytic  leukocyte
counts,  differential  granulocytic  leukocyte  counts,  reticulated  erythrocyte
counts,  basophilic aggregation  estimates,  platelet  counts, erythrocyte  sedi-
mentation  rates, coagulation time, hematocrit values, erythrocyte fragility, or
serun  bil"irubin levels.   Approximately 75%  of  the  painters were exposed  to
concentrations of 500 ppn. or less, and the group had  no known prior exposure to
benzene.   However,  the  contamination  of the toluene  vehicle in  the paint  with
benzene cannot be precluded  (NIOSH,  1973), because these blood  changes are con-
sistent  with  those of  benzene  poisoning.   Volatile components such as  ethyl
alcohol, ethyl acetate, butyl alcohol, and petroleim naphtha  were also present in
quantity in the lacquers, dopes, and  brushes used by  the workers (Table  11-9).
     In  19^3,  Wilson  reported that  of approximately  1000  industrial  workers
(industry not stated) exposed to 50 to 1500 ppm of  commercial toluene vapor for 1
                                     11-22

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                                   TABLE  11-8

 Results of Blood Examinations Performed on Toluene-Exposed Airplane Painters3'
                                        Toluene-Exposed
                                            Workers
                      Unexposed
                       Workers
Erythrocytes       ,
   counts <5.2 x 10 /mm
Lymphocytes
   counts >5000/mm
Mean Corpuscular Volume
   >100 ^
Hemoglobin
   _>l6g/100 mJi

Mean Corpuscular Hemoglobin
   >35 picograms

Mean Corpuscular Hemoglobin
   Concentration
   % of cases 2.35g/100 mfc
13.1$ (N = 61)
20.^ (N = 59)
34.4$ (N = 61)
5.2? (N =
1.1% (N = 395)
21.3? (N = 61)      7.2$ (N = 111)
29.5? (N = 61)      2.4? (N = 81}
13-1? (N = 61)      0? (N = 73)
2.5? (N - 81)
 Source:  Greenburg et al., 1942
 Percent abnormal cases reported.
                                         11-23

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                              TABLE  11-9

                  Analysis of Paint Used by Painters3
                                             Percentage
                                             in Mixture
Spray painters
  Primer (75$ of paint used):
     Zinc chromate
     Magnesium silicate
     Synthetic resin
     Driers (lead and cobalt compounds)
     Xylene
     Toluene
   Nonvolatile:
     Resin, titanium oxide, zinc oxide,
     ultramarine blue, ferrocyanide
     blue, iron oxide, diatomaceous
     earth, amorphous silica, carbon
     black
                                               100.0
  Lacquer 1 (15$ of paint used):
   Volatile portion:
     Ethyl alcohol                               7.0
     Ethyl acetate                              18.0
     Butyl alcohol                               7.0
     Butyl acetate                              15.0
     Petroleum naphtha                           3.0
     Toluene                                    50.0

                                               100.0

   Nonvolatile:
     Nitrocellulose, synthetic resin,
     titanium oxide, ferrocyanide blue,
     iron oxide, carbon black, zinc oxide,
     etc.  No lead compounds

  Lacquer 2 (10$ of paint used):
   Volatile portion:
     Toluene                                    25.0
     Xylene                                     33-0
     Petroleum naphtha                          ^2.0
                                               100.0

-------
                          TABLE 11-9  (cont.)
                                             Percentage
                                             in Mixture
Brush painters
  Dope:
   Volatile portion:
     Ethyl acetate                              16.5
     Ethyl alcohol                               3.2
     Butyl acetate                              16.5
     Butyl alcohol                               5.6
     Pttroleum naphtha                          13,7
     Toluene                                    114.5
                                               100.0
   Nonvolatile:
     Nitrocellulose, glycol sebacate,
     aluminum, cadmium sulfide, barium
     sulfate

Brush wash:
     Acetone
     Ethyl alcohol
     Toluene
                                               100.0
aGreenburg et al.,

 Dip painters used a primer only of the same composition as given
 for spray painters.
                                     11-25

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to 3 weeks, 100 showed symptoms attributable to toluene intoxication.  Ten of the
100 workers had been exposed to concentrations in excess of 500 pprn and showed
signs of serious CNS toxicity  (Section  11.1,1.2.).   In most of these 10 cases,
all  blood  elements remained  normal  except for  the  red cell  count,  which  was
"usually" reduced  (=2.5 x 10 /mm^).  in  2  of  the 10  cases,  leukocytes (2500 to
3000/mrr) and  platelets were reduced  as  well, and  differential  counts showed
decreased polymorphonuclear  cells  and reticuloeytes,  and  increased monocytes.
Sternal bone marrow  biopsies  in these two cases showed partial degeneration of
the blood-forming elements, which resulted in a diagnosis  of aplastic anemia.  No
clinical blood changes were seen in the workers  who had been exposed to the lower
concentrations of toluene  (i.e., <500 ppm) .
     Von Oettingen et al .  (19423,  19M2b) were the first workers to document the
effects of  essentially  pure  toluene on human  subjects.   The  toluene  used was
shown, on spectrophotometric  analysis, to contain not  more than 0.01J  benzene.
In this study, no  significant changes in the total  or differential  white cell
count were found in 3 volunteers  following controlled 8-hour exposures to various
concentrations of toluene  within the  range  of 50 to 800 ppm.   Not  more than  2
exposure sessions were performed per week to provide sufficient  time for recovery
between exposures, and  the experiments were  conducted  over a period of 8 weeks
(Section 11.1.1.1.).  Erythrocyte  counts were not made.
     Parmeggiani and Sassi  (195^) concluded from a clinical study of 11 paint and
.pharmaceutical workers exposed to 200  to 800  ppm toluene and 13  others with expo-
sure to a  combination of  toluene  (150 to 1900 ppm)  and butyl  acetate  (150  to
2^00 ppm) that toluene had no particular  injurious action on  the bone marrow (or
other organs). The English summary of this study indicated that the workers were
exposed for "many" years,  but the purity of  the toluene was not  reported.  Among
the workers in the two  groups,  3^? reportedly showed slight  anemia (2000/mm ), and 45J  showed a decrease in  blood platelets (<150,000/mmJ)
not accompanied by evident signs of capillary fragility.
     In a  more recent  investigation,  Banfer  (1961)  examined  889  rotogravure
printers and helpers who were exposed  to the  vapors of  toluene-containing print-
ing  inks  for at  least   3  years.   Four hundred and  seventy eight  non-exposed
persons from two groups  served as controls; one  group was composed of 155 manage-
ment workers  from the  same  plant, and  the  second  group  was   composed  of 323
persons from outside the plant.  The available  commercial toluene used in these
                                     11-26


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 inks reportedly  contained  only  traces  of  benzene  (<0.3$);  when  5  samples  of  the

 toluene were  examined by Banfer, no traces of benzene were found,  but the  method

 of  analysis and  detection  limits were  not stated.  Analysis  of  the  room air  for

 toluene was  performed by infrared spectroscopy, but limited to 5 samples taken

 from different sites.on a  single day.   Ambient toluene concentrations were  not

 specified, but three  of the  samples were  determined  to be  below the "MAK-Wert,"

 the fourth sample was at the "MAK-Wert," and the fifth  sample, taken near  one of

 the presses,  exceeded the  "MAK-Wert" by 400 ppm.  A translation of this study by

 NIOSH  (1973)  indicates that  the  "MAK-Wert"  was 200 ppm.   Hematologic  examina-

 tions  of  the  workers  and controls  did  not roveal  any significant  changes  in  the

 total  number  of leukocytes, lymphocytes, granulocytes,  or  erythrocytes,  or hemo-

 globin levels (Table 11-10).  Sternal biopsies from 6 printers with white cell

 counts of less than 5000/mm   were  normal.


                                   TABLE 11-10

               Henatologic Examination  of  889  Rotogravure Workers3
Printers
(N = 889)
Controls,
Group 1
(N = 155)
Controls,
Group 2C
(N = 323)
Erythrocytes     ,
   counts <4 x 10 /mm-1
Leukocytes, total
   counts > 8500/mo
   counts <5000/mm
   counts
   counts <4000/nm-3

Lymphocytes
   <35$ total leukocytes.
   total counts <5000/mm:

Granulocytes __
   total counts >2000/mnr

Hemoglobin
   value <13g/100mJi,
16 (1.79$)
78 (8.77$)
74 (8.32$)
28 (3.15$)
 3 (0.33$)
                                        25 (2.81$)
                                       889 (100$)
                                                       3 (1.93$)
               7  (2.10?)
11  (7.09$)     26  (8.04$)
18  (11.61$)    38  (11.76$)
 4  (2.58)      12  (3-71$)
 1  (0.64$)      1  (0.30$)
               3 (4.16$)
             155 (100$)
                                                       4 (2.58$)
               4  11.32$)
             323  C.00$)
                                       889 (100$)    155 (100$)     323 (100$)
                             4 (1.23$)
 Source:   Banfer, 1961

 Unexposed management workers from the same plant
Q
 Unexposed individuals not enployec! at the plant
                                      11-27

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                               I
     Capellini and  AjLessio  (1971)  performed hematological  examinations  on 17
workers who had been exposed for "diverse" years  to 125  ppm toluehe  (range, 80 to
160 ppm) in  a plant manufacturing  V-belts  for industrial  machinery.   Results
showed  that  the  hemoglobin  values, red  cell  counts,  white cell  counts,  and
platelet counts of  the workers  were within  the same limits as those of 19 non-
exposed control subjects  from the same plant. The  benzene content of the toluene
was  not reported.    Blood findings  were  also  within  normal  limits  in another
worker employed in a different department who was exposed to mean toluene concen-.
trations of 250  ppm (range, 210 to 300  ppm)  and  who demonstrated symptoms, of CNS
toxicity and conjunct!val irritation.
     In 1975, a report by the West German Association of Gravure Printers (Suhr,
1975) identified a  study population of 100  printers with at least 10 years of
exposure to pure toluene (<0.3% benzene) and an  unexposed control group of equal
size from the same plant.  Analysis  of air samples collected from the workplace
reportedly indicated that  the  potential  exposure  to toluene ranged from 200 to
400 ppm.  Blood analyses  (hemoglobin, erythrocyte,  leukocyte, thrombocytes, dif-
ferential analysis) demonstrated no unusual  frequency of abnormalities in either
the exposed or control groups.
     Matsushita et  al.  (1975) found no  alterations  in the  specific gravity of
whole  blood,  hemoglobin  content,  ftematocrit, or  white blood cell  counts  in a
group  of  3& female shoemakers  who had been exposed to toluene  (60 to 100 ppm
average) and, in a  "few" working  places,  gasoline (range,  20 to  50 ppm) for an
average duration of  3 years  and 4 months.  Tne hematological test results from
the shoemakers were  compared with tho^e from an  unexposed  control  group  of 16
female  workers.  A  significantly increased number  of "Mommsen's" toxic granules
were observed in  the neutrophils  of the  exposed  workers.   Thirteen  of  the 38
Workers showed an abnormal  appearance  of the granules  (mean number per neutro- .
phil, 7.6 +_ 5.6)  compared with 1 of 16  controls  (mean number per neutrophil, 3.8
t  3.4).
     Further evidence of the relative non-toxicity of toluene to the hematopoie-
tic system was presented by Francone and Braier  (1954). Toluene, because of its
presumed myelotoxic action, was administered orally as  a treatment  for leukemia.
It was  found that daily  doses of up to  10 g  of  toluene in olive oil for 3 weeks
(to a total of 130 g) were tolerated by leukemia  patients without complaints or
evidence  of side  effects,   but  the treatment  had  ho clinical  effect  on the
leukemic process.
                                      11-28

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     Hematological  abnormalities  have  been infrequently reported in sniffers of
toluene-based  glues.   In a total of 90  cases  surveyed by 4 groups  of  investi*
gators  (Christiansson  and Karlsson, 1957; Kassengale et al.,  1963; Barman et al.,
1964; Press and Done,  1967b),  there were  no instances of anemia or lymphppenia,  a
single  report  of neutropenia, and  6  cases of eosinophilia  of  greater  than 5%.
Christiansson  and .Karlsson  (1957) aldo performed bone marrow examinations on 17
individuals; 10  of  these showed changes  suggestive of disturbances in maturation
of leukocytes, although these changes were not reflected in the peripheral blood
of the  same  individuals.   The  individuals  examined in the Christiansson and
Karlsson (1957)  study were habituated to the  inhalation  of  toluene-based paint
thinners,  rather than model glues as  were  the subjects in other surveys.  In a
fifth clinical survey of 89  glue sniffers, however, Sokol  and  Robinson  (1963)
found abnormalities of the blood in 68  of the cases.  An effect on the white blood
cells was  indicated by findings  of  eosinophilia  (25 subjects),  leukocytosis (12
cases), and lymphopenia (4 subjects).   Sokol  and  Robinson (1963)  also  reported
low hemoglobin values  in 20 subjects and basophilic stippling of erythrocytes in
42 of  the patients,   and  noted   the frequent  occurrence of  poikilocytosis (25
                                                                      i
cases), anisocytosis   (20 cases), hypochromia  (14  cases),  and  polychromasia (10
cases). There is  no obvious  explanation for the discrepancy between the hemato-
logical findings of Sokol and Robinson  (1963) and those of the other  investi-
gators.  However,  since none of  the aforementioned cases deal  with exposure to
pure toluene,  the abnormalities observed should  be considered to be the  possible
result of  contamination of-the toluene by benzene or some other organic  solvent.
     Powars (1965)  diagnosed  five cases  of acute aplastic anemia that were asso-
ciated with glue  sniffing in black  adolescents with  pre-existing sickle-cell
disease.  The 5 children  had apparently used  3 different  glues,  2 containing
toluene and 1 containing  acetone.  All of these  patients  recovered following
transfusion and  cessation  of  sniffing.  A case of fatal aplastic anemia,  uncom-
plicated by the  presence of sickle-cell disease, was described in a sixth indi-
vidual  with a  3-year  history  of  glue  sniffing.
11.2.2.   Blood  Coagulation.   Pacseri  and Emszt  (1970;  cited  in  NIOSH,   1973)
reported that  an increase  in  prothrombin time was found in 191 printers exposed
to 170  to  340  ppm toluene (duration of exposure not stated). Two of  the subjects
showed a reduced number  of  red  blood  cells, but  no other hematologic abnormali-
ties were found in these  workers.   The  benzene content  of  the toluene was not
reported.
                                     11-29

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  \

11.2.3.   Phagocytic  Activity of  Leukocyt.es.    It  has  been  reported that  the
phagocytic  activity of leukocytes  from printing-plant workers exposed to toluene
vapors  was  significantly  reduced relative  to a  control population  (Bansagi,
1968).   There was  no relationship, however, between the decrease in activity and
the concentration  of toluene in the air.  The Chemical Abstracts summary of this
Hungarian study did not detail  any  of the exposure  information  or  mention the
benzene  content of the  toluene.
     Friborska  (1973;  cited  in  NRC,  1980) noted  increased concentrations  of
alkaline phosphatase  and lactic acid dehydrogenase in leukocytes  and  increased
acid phosphatase in both leukocytes and lymphocytes  from workers  who  were rou-
tinely  exposed  to toluene.   The authors  associated  these  alterations  with
increased functional  capacity of the  cells.
11.2.4.   Immunoeompetence.  Serum  immunoglobulin level (Lange et al.,  1973a) and
leukocyte agglutinins (Lange et  al., 1973b) were studied in a group of 35 workers
with a  history  of exposure to benzene; toluene, and  xylene.   The duration  of
exposure ranged from  1  to 21  years and the  concentration of  these compounds  in
the air  ranged from 0.011 to 0.17 mg/Jl, 0.08 to 0.23 mg/2,, and 0.12 to  3.0  mg/£,
respectively.  Serum  IgG  and  IgA levels were found  to be significantly lower  in
the solvent-exposed workers than  in  non-exposed controls, although IgM levels
tended to increase (Lange et al.,  1973a).  Lange and coworkers (1973b) also  found
that 10-of the 35 workers had leukocyte agglutinins  for  autologous  leukocytes,
and demonstrated  an  increase  of  leukoagglutination  titer in  human sera  after
incubation with benzene,  toluene or  xylene;  this  suggested  that some  workers
exposed simultaneously to these aromatic compounds may  exhibit allergic  blood
dyscrasias.  In another  group of workers  (N  = 79)  with a  similar history  of
exposure to benzene, toluene, and  xylene (i.e., levels and durations of exposure
comparable to  those  of  the  workers examined  by Lange  et al.),  Smolik et  al.
(1973)  found a decreased  level  of serum complement.  It  should  be  noted that  in
all of the aforementioned studies, the specific solvent(s) responsible  for the
changes  were not identified.
11.3.  EFFECTS ON  THE LIVER
     Greenberg et  al. (1942) found enlarged livers in 13 of 61 airplane painters
(21J) who were  exposed to  100 to 1100 ppra toluene for  2 weeks to more than  5
years.   Toluene was the major solvent used in the  paints, although significant
quantities  of other volatile components were present (Table 11-9);  these workers
reportedly had  no  history  of inhalation  exposure  to any other  toxic volatile
                                     11-30

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solvents,  including benzene.  This  incidence of liver enlargement  was  3 times
that observed in  a  control  group of 430 workers who  had never been exposed to
toluene1, but  it  cannot  be  correlated with  exposure  level,  because only  the
numbers of workers  exposed  at different exposure levels (and not hepatomegaly
incidences) were reported.  The liver enlargement was diagnosed by palpitation,
and in no cases were the  livers  tender.   There was also no correlation between
the enlarged livers  and either clinical or laboratory  (blood and urine analyses)
evidence of disease, and  it was  suggested  that the enlargement might have been
compensatory in nature.
     Greenburg and coworkers'  ( 1942) finding of  hepatomegaly  has  not been sub-
stantiated in subsequent studies  of workers with  histories of  occupational
toluene exposure.   Parmeggiani  and  Sassl  (1954) found  a  comparable incidence
(21%) of enlarged livers  in a group of 11 paint and  pharmaceutical projection
workers who were  exposed to 200 to  800 ppm  toluene  for  "many"  years,, and in a
control group of unexposed workers from the same plant.  Normal liver function,
as determined by electrophoresis,  serum  colloid stability testing, and galactose
tolerance testing,  was also  observed in  the  exposed workers.   Capellini  and
Alessio (1971) observed no changes in "the function of the liver"  in 17 workers
exposed for  "diverse"  years  to  a  mean atmospheric  concentration  of  125 ppm
toluene (range, 80  to  160 ppm)  in a plant  manufacturing V-belts for industrial
machinery. Liver function in this study  was evaluated  by  determinations of total
serum protein and protein electrophoresis.
     M-ore recently, Suhr (1975)  similarly found comparable, but high, incidences
of enlarged livers and elevated liver enzymes in a group  of 100 gravure printers
with at least 10 years' exposure to  200  to  400 ppm  pure toluene (benzene 
-------
investigated.   Blood  .alcohol determinations  before arid  after the  workshifts
indicated comparably elevated levels in both the printers  and control group, but
less than half of the 100 subjects in each group were test-^J;  approximately half
of the tested subjects had levels between 0,01 and 0.1J. The significance of the
elevated blood alcohol levels is unclear,  however,  because  of the small number of
subjects tested, because only single biood alcohol del.erminations were performed
on each subject, and because the data  were presented ambiguously.
     Other studies  have reported significant effects on indices of liver func-
tion in groups  of  toluene-exposed workers.  In  an examination of 9^ rotogravure
printers with a history of exposure to 18 to 500 ppm toluene and of a reference
group of 30 municipal clerks, Szadkowski et al.  (1976)  found significant reduc-
tion  in  bilirubin  and  alkaline  phosphatase  in the exposed group,  but  no dif-
ference from controls in  SCOT,  SGPT,  leucinamino-peptidase, or  cholinesterase
levels.  The 9^ rotogravure workers were categorized into four  groups by inten-
sity of exposure to toluene.  The mean exposure  levels, durations of exposure and
ages  of  the groups were, respectively  (Szadkowski  et  al., 1973):   Group 1 (N =
'68)  - 300 ppm,  7.3 +  5-3 years,  32 years;  Group  2 (N =  4)  - ^26  ppm,  newly
appointed on day of investigation, 2^.3 years;  Group 3 (N - 11) -  82 ppm, 5.6 +_
5.2 years, ^2.9 years;  Group 4  (N - 11) - 18 ppm,  8.5  +_  ^.4 years, 35.8 years.
Blood alcohol levels ranged from 0.02? to 0.07$ in the exposed  workers.
     Trevisan ana Chiesura  (1978)  performed the  following  hepatic function tests
on  ^7 subjects who  were exposed  occupationally  to  toluene  via inhalation:  bili-
rubin,  SCOT,  GGT,  alkaline  phosphatase  (AP).ornitnine-carbamyl  transferase
(OCT.), Uuick's test, and protein measurement.  All tests gave normal results with
the exception of GGT,  which was reportedly above normal (28 n/m£)  in 3^% of the
cases.  In a group of  12 subjects controlled  before  and after toluer.e entered in
the  working  operation, mean GGT  activity  increased  2-fo]d  after  exposure.
Although  GGT  has  proved  to be  a  very sensitive  screening enzyme  for  slight
changes in liver function  (Dragoslcs et al.,  1976), it should be noted that the
data  from  this study were published in abstract  form, and that  information on
exposure or type of occupation and detailed results of the  hepatic function tests
were not presented.
     The mean serum  activities  of four liver  enzymes  (aspartate  transaininase,
alanine aminotransferase,  OCT,  GGT) did  not  differ  between a  group  of  102 car
painters who were exposed to a  mixture  of organic solvents, and an age-matched
unexposed reference group  of 102 men  (Kurppa  and  Ilusman, 1982).    The  exposed
                                     11-32

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   \

subjects were exposed mostly to low levels of toluene (30.6 ppra detailed in Table
11-3)  for  1 tO HO years (mean, 14.8 •«• 8.5 yrs), and the  age of the painters ranged
from 20 tO 65 (mean 35 +_ 11 yrs).  Abnormal  intellectual/psychomotor performance
(Hanninen  et al.  1976), abnormal neurophysiological effects (Seppalainen et al.,
1978)  and  an increased frequency of lens changes (Raitta et  al.,  1976), however,
have   been  observed  in  thf.se  workers.    Abnormally  slow motor  and  sensory
conduction velocities and/or  prolonged motor digital latencies were suggested in
12 of  ^9 car painters studied  (Section 11.1.2), and ophthalmological examination
revealed  lens opacities  in 48 of 92 car painters  (Section  11.7.1).   Kurppa and
Husman  (1982)   found   that,   the   liver   enzyme   activities   of   these   two
"solvent-affected" subgroups were similar to those of  the  car  painters with no
corresponding  abnormalities.
     English summaries  of two Polish studies of women with  histories of occupa-
tional exposure to toluene  indicated  abnormalities in the glycoprotein,  serum
mucoid and haptoglobin patterns of 53 women  (Kowal-Gierczak  et  al.,  1969), and
changes in  the serum  levels  of  iron  and copper,  and urinary  excretion  of
porphyrin in 51  women (Cieslinska  et al.,  1969).  Although  these changes may be
indicative of liver  dysfunction,  clinical signs  of liver function  impairment
were not  observed in  these subjects.  The concentrations of  toluene, durations of
exposure,  and  the possibility of exposure to other chemicals  were  not discussed
in the available summaries.
     Intensive  exposure  to toluene via glue or thinner sniffing appears  to have a
minimal effect  on the liver.   Results of hepatic function tests .(SCOT, SGPT, AP,
bilirubin, sodium sulfobrotnophthalein excretion, serum proteins, cephalin floc-
culation) on a  total  of  179 sniffers who were examined in early clinical surveys
were  essentially  unremarkable  (Christiansson .and  Karlsson,  1957;  Massengale
et al., 1963;  Sokol  and  Robinson,  1963; Bam an et  al.,  196-'J;  Press  and  Done,
 1967a, 1967L).  Christiansson and Karlsson (1957) apparently  did  detect  liver
enlargement in  5 out  of  32 Swedish lacquer thinner sniffers,  but other signs of
liver  function  were  normal.   More recently,  Litt and coworkers  (1972)  found
elevated SGPT  and AP levels  in  2  and  5$,  respectively, of  a group  of  982 glue
sniffers.
     Grabski (1961) described an individual who had abused  pure toluene for six
years  and showed signs of  cerebellar   degeneration,  hepatomegaly,  and impaired
liver function.  Complete series of liver function tests were normal, however, in
an optometrist and a glue sniffer exposed independently to 99? pure toluene, both
                                     11-33

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Of whom  alSo  exhibited encephalopathic effects (Boor and Hurtig, 1977).  Rever-
sible hepatorenal  damage was  diagnosed in  an individual with a 3-year history of
inhaling a cleaning fluid that contained 80? toluene  (other components not known)
coupled  with alcohol  ingestion (O'Brien  et  al,  1971);   the hepatic  effect was
indicated  by  elevated serum bilirubin and AP.
11.1*. EFFECTS  ON  THE KIDNEYS
     Urinary  findings were normal  in 91 specimens  from  a  group of 61  airplane
painters (number  of  donors  not  stated) who  were  exposed to  100 to  1100 ppm
toluene  for 2 weeks to 5  years  (Greenburg et al.,  1942).   Urinalysis consisted of
specific gravity,  albumin,  and  sugar determinations, arid examinations for formed
elements.   Exposure  to  mean  concentrations of 60 to  100 ppm  toluene and 20 to
50 pptn gasoline in a  "few" working places for an average duration of 3 years and
4 months did  not result in abnormal urinalysis findings as determined by standard
methods  (protein,  sugar, urobilinogen,  bilirubin, occluded blood,  keton body),
except  for  excretion of hippuric acid,  in  38  female  shoemakers  (Matsushita
et al.,  1975).   Gloraerular filtration  rate  (as measured  by   Cr-EDTA clearance
from plasma)  was not  reduced in a group of 3^ rotogravure workers when compared
with 48 non-exposed  male controls (Askergren et al.,  1981),  but the toluene
exposures   were not   characterized.    Proteinuria  and   hematuria  were  noted,
however, in a worker  who was exposed  to concentrations of toluene sufficient to
cause unconsciousness while cleaning  the inside of a tank that  was coated with an
emulsion of 45J toluene and 21% DDT (Lurie, 1949).
     Reisin and coworkers (1975)  published a report  regarding the development of
severe myoglobinuria  and n:>n-oliguric acute  renal  failure in a  paint factory
laborer who  was exposed  to pure toluene  by skin contact and  aspiration when a
hose burst.  The  patient had inhaled  sufficient  amounts of toluene  to cause a
loss of  consciousness for 18 hours and subsequent  development of chemical pneumo-
nitis and  sustained superficial burns  on  approximately  10$ of his  body surface
area.   Acute renal  failure apparently developed from  the  lack of  fluid intake
accompanied by heavy  myoglobinuria rather than from a direct  effect of toluene.
The  early administration of intravenous  fluids  and diuretics, and  the use of
hoaodialysis  led to  complete recovery.
     Pyuria,  hematuria,  and  proteinuria have  been  the most frequently observed
signs of renal  dysfunction  associated  with the deliberate inhalation of toluene-
based glues  (Christiansson  and  Karlsson, 1957; Massengale et al.,  1963; Sokol and
Robinson,  19&3; Barman et al., 1961; Press  and  Done,  1967a,  1967b).  The clinical
                                     11-34

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findings observed in  159  cases surveyed between 1957 and  1967  are  tabulated in
Table 11-11.   These indications of renal dysfunction have  not  been universally
observed in glue sniffers, are generally transient, and follow closely the inten*-
sive exposures  (Press  and Done,  196?b).
     0'Brj°n  et  al.  (1971) more  recently described a case of reversible hepato-
renal damage  in a  19-year-old male  who had a  3-year  history of glue  sniffing
while employed in  the  sign-painting trade.   Prior to  hospital  admission,  the
subject had spent  6 hours inhaling a cleaning  fluid that  contained 80? toluene
(the other components  were not identified).   Upon admission, the patient  was
vomiting and  anuric, and  after  8  hours, periorbital edema  and 'subconjunctival
hemorrhages  developed.   Blood  concentration  of toluene  was determined  to be
160 ppjo.   In  addition to  diminished  urine  output, evidence  of  renal  damage
included hematuria,  proteinuria, and elevated serum creatinine.  The effects of
these exposures  on  hepatic function are discussed in Section 11.3. (Effects on
the Liver).
     Although serious  involvement  of   the  kidney with human intoxication by
toluene has not  been stressed  in  the   early  literature,  several  reports  have
recently appeared that  associate deliberate inhalation of toluene with metabolic
acidosis (Taher  et  al., 197^; Fischman  and Oster,  1979a;  Kroeger et al.,  1980;
Bennett and Forman,  I960;  Moss et al .,  1980).  The cases  of acidosis described '.j
these  investigators  (Table  11-12)  are  characterized   by  serious  electrolyte
abnormalities  (hypokalemia,   hyperchloretnia),   and  are related   primarily  to
toluene's ability to impair  hydrogen ion  secretion in  the  distal  renal  tubule
 (distal renal tubular acidosis). In addition to findings compatible with distal
renal tubule  acidosis, Moss et al.  (1980)  found pathologically increased excre-
tion of amino acids, glucose, phosphate, uric  acid,  and calcium  that indicated
proximal tubule  dysfunction consistent  with Fanconi's syndrome.  Kroeger et al.
 (1980) reported  the  case of a patient with toluene-induced renal tubular acidosis
who  developed recurrent urinary calculi.  It  should be noted  that  each  of the
subjects who  developed acidosis had a history of multiple  toluene abuse and,
although the  acute  consquences of renal  tubular acidosis associated with toluene
sniffing were on occasion life threatening, these effects were completely rever-
sible  with abstinence from  toluene  exposure.    These   symptoms  also responded
promptly to  electrolyte  repletion therapy  with potassium  chloride  and  sodium
 bicarbonate.
                                     11-35

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                                                    TABLE 11-11
                                 Renal Funqtion Investigations  of Glue Sniffers
                                                                               a,b
Number of
Patients
32
27
89°
15
16
Pyuria Hematuria
All 32 urine ND
samples "normal";
details not given
0 2
32 14
0 0
6 3
Proteinuria
ND
0
12
1
5/13
Clearances
ND
ND
ND
PSPd
0/13
Urea
1/7
Azotemia
ND
0
ND
0/7
0/9
Reference
Christiansson and
Karlsson, 1957
Massengale. et al.,
1963
Sokol and Robinson,
Barman et al. , 196U



1963

Press and Done, 1967b
 Source:  Press and Done, 1967b
 Exposures were to toluene-containing plastic cements  except  in  the  Christiansson and Karlsson  (1957) study,
 in which the subjects examined had sniffed paint thinner.
 Urinary abnormalities were found in 67 of the 89 glue sniffers.
 Pnenosulfonphthalein clearance in 2 hours.
ND = not determined

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                                                                          TABLE 11-12

                                                              Toluene Induced Metabolic Acidosis
      Subject  (Age)
                               Exposure History
                                                                      Symptoms
                                                                           Clinical Findings
                                                                                                                                     Reference
      Kale (23 yr)




      Feoale (20 yr)



      Female (17 yr)


      Feaal« (21 yr)
Z.    Fco4l« (25 yr)
 I
CO
~j
      Hale (23 yr)
      Fenale (27 yr)
      Four Indlvlducls
      (ages and sexes
      not stated)

      Male (2? yr)
 Sniffed glue and pure toluene
 Intermittently for 6 yr.
 Two 3  to  5  d  episodes  of  sniffing
 aerosol paint containing  60)
 toluene within 
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     Fischman  and Oster (1979a) found a  high  anion gap metabolic acidosis with
hypokalemia  in two  patients  who had  sniffed  100$  toluene;  this  condition is
reportedly indicative of an increased production of acid by the body.  Although
it was noted that renal  failure,  ketonemia, and elevated  lactate  levels could
have accounted in part for the  abnormal  increases in an:on  gap, it was suggested
that the  acid metabolites  of toluene  (e.g.,  benzoic and hippuric acids) may have
caused the high anion gap metabolic acidosis.
     Clinical  manifestations  associated with the reported metabolic alterations
included   nausea,    lethargy,   ataxia,   muscular   weakness,   and   paralysis
(Table 11-12).    The National  Research. Council has  noted that  some  of these
manifestations may mimic those usually  attributed  to  the effects of toluene on
the CMS,  and that altered pH and electrolyte  balance may  be more commonly respon-
sible for the manifestations of toluene  abuse than is usually recognized (NRC,
1980).  In particular, hypokalemia often produces significant muscular weakness
including flaccid paralysis.
11.5.  EFFECTS ON THE HEART
     Ogata et  al.  (1970) found an apparent decrease  in the pulse  rate of 23
volunteers who were exposed to  200 ppm  toluene for periods  of  3 hours,  or for
3 hours and  a 1-hour break period  followed by 4 additional  hours,  but no effect
at 100 ppm.  Systolic and diastolic blood pressure were not  affected by exposure.
Exposure to 100  and 200 ppm  toluene for  30 minutes did not, however,  have any
effect on the  heart rates or  electrocardiograms  of  15 other  subjects  during
either rest  or light exercise (Astrand et al.,  1972).   Other studies have shown
that experimental exposure to toluene  at  levels of  100 to 700 ppm for 20 minutes
(Gamberale and Hultengren,  1972)   or  50  to 800 ppm for  8  hours  (Von Oettingen
et al., 19^2a,  19l;2b) did not cause any definite effects on heart rate or blood
pressure. Suhr (1975) noted that  the pulse rates and  blood pressures of  a group
of 100 printers with a 10-year  history of exposure to 200 to 100 ppm toluene and
those  of  an unexposed  control group  of  identical  size  were  similar  at  the
beginning and end of work shifts.
     Sudden  deaths   that  were  not  due \.o suffocation  secondary  to   solvent
sniffing, but rather were attributed  to a  direct  effect of  the solvent itself
have  been reported  in  at least   122 cases (Bass, 1970;  AIha et  al.,  1973).
Toluene,  benzene,  arid gasoline have  been  individually implicated  in  a small
number of these  deaths  (10,  6, and  H  cases,  respectively), but  the volatile
hydrocarbons most   frequently  involved  were  trichloroethane  and  fluorinated
                                     11-38

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aerosol propellants.   Severe  cardiac  arrhythmia resulting  from  light  plane
anesthesia was offered as.the most likely explanation for the cause of the sudden
sniffing deaths.  Bass et al.  (1970)  noted  that stress, vigorous activity, and
hypoxia, in combination with sniffing, appear to increase the risk of death.
11.6.  EFFECTS ON MENSTRUATION
   i  Subjective complaints of  dysmenorrhea were  reported by 19 out of 38 Japanese
female shoemakers (mean age, 20.7 years) who were exposed to mean  toluene concen-
trations  of  60  to  100 ppm for  an  average duration  of 3 years  and 4 months
(Matsushita et al.,  1975).  In an vinexposed control  group of 16 women from the
same plant, this  effect  was noted  by  3 individuals  (19J).   It  should be noted
that these women  were concomitantly exposed to 20 to  50 ppm of gasoline  in a
"few"  working  places.  In  this study,  the  presence or  absence of  15 subjective
symptoms  was   ascertained  by   questionnaire;  in  addition  to  dysmenorrhea,  a
significant  number  of  workers  reported  "uneasy  feelings"  about  the  solvent
vapor;, and itching and dermatitis of the hands.
     Michon (1965) reported disturbances of menstruation in  a group of 500 women
 (age 20 to 40  years) who had been exposed to a  mixture  of benzene,  toluene, and
xylene in  the  air of  a leather and rubber shoe factory.  The concentration and
component  distribution of this mixture were not specified, but it  was stated in
the  English summary of this Polish study  to be within  permissible occupational
limits established at the time in Poland, 31 ppfli (100  mg/m )  for  benzene, 67 ppm
          o                                   •?
 (250 mg/m-5) for toluene, and 58 ppm (250 mg/m^)  for xylene).   When  the menstrual
cycles of  the  exposed women were compared with  those  of 100 w\nen  from the same
plant  with-  no exposure  to  these  hydrocarbons,  prolonged  and  more  intense
menstrual  bleeding was reportedly found in  the exposed group.  The  regularity of
 the  cycle  was  not affected.
     It  has  a] so been  noted   in  the English summary  of a  Russian study that
cccupational exposure to average concentrations  of 6  to 93 ppro (25  to  350 mg/m  )
 toluene and other solvents, through  the use of  organosiliceous  varnishes in the
manufacture  of  electric  insulation  materials,  caused  a  high  percentage  of
menstrual  disorders  (Syrovadko,  1977).    The newborn  of  these  women  were
 reportedly more  often underweight  and experienced more frequent fetal asphyxia
 and  "belated"  om.et of nursing.  Although the above studies  suggest that occupa-
 tional exposure  to  aromatic  hydrocarbons  may  be  associated  with menstrual
 disturbances,  it should be emphasized that  a specific effect of toluene could not
                                      11-39

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be determined from the available data,  Information on the possible reproductive
effects  of  toluene  in males is not available.
11.7.  EFFECTS  ON THE RESPIRATORY TRACT AND THE EYES
11.7.1.   Effects of Exposure.   Carpenter et  al.  (1944) observed  that  2 male
subjects who were exposed to toluene for 7  to  8 hours experienced transitory rHd
throat and  eye  irritation  at  200  ppm,  and lacrimation at 400 ppm.  Parmeggiani
and Sassi (195*0 found irritation  of the upper respiratory tract and conjunctiva
in 1 of 11  paint and  pharmaceutical  product  workers who were exposed to 200 to
800 ppm toluene  for "many"  years.   In  the studies  of Von Oettingen  et  al.
(1942a,b)   and  Wilson  (1943),  however,  no  complaints  of  respiratory  tract
discomfort  were recorded in volunteers or workers exposed to  levels of toluene as
high as  800 to  1500  ppm for 8-hour periods (Section 11.1., Effects  on the Nervous
System). In 2  episodes of  accidental poisoning on ships that involved estimated
short-term  exposures  to  10,000  to 30,000 ppra  toluene,  Longley et  al.  (1967)
recorded no complaints of respiratory tract or eye irritation among 26 men.
     Transient  epithelial injury to the eyes  that consisted  of moderate conjunc-
tiva! irritation and  corneal  damage,  with no  loss  of vision,  was  observed in
three workers  who were accidentally splashed with  toluene   (Mclaughlin, 1946;
Grant, 1962).   Complete recovery generally occurred within 48 hours.  The results
of opthalmologic  examinations  of 26 spray  painters who were exposed to toluene at
levels of  100 to  1000 ppm for 2 weeks  to  more  than  5  years  were reported to be
negative'(Greenburg cc al., 1942); results were not published, but  it was noted
that the examinations in  each  case consisted  of a "history of ocular complaints,
visual  acuicy,  fundus,  pupil and  slit lamp investigation  of  the media  of the
eye."
     Raitta and coworkers (1976,' found  lens changes  in a group of 92  car painters
who were exposed  to a mixture of organic  solvents for  1  to  40 years (mean 15 +
9 years).   Of  the  organic solvents  detected  in  the  breathing  zones  of  the
workers, toluene was present in the greatest amounts  (30.6 ppm);  the  mean concen-
trations of the other solvents present in the air are  included in  the summary of
the Hanninen study   (Table  11-3).  This study was part of a large investigation
performed  to evaluate  the effects  of  chronic solvent  exposure on  the nervous
system  (Hanninen et al.,  1976; Seppalainen  et  al.,  1978)  and  liver function
 (Kurppa and Husman,  1982)  of the  car  painters (see Sections 11.1.   and  11.3).
Among the 92 car painters  (mean age 34.9  + 10.4 years, range 21 to  64 years),  2
had  been operated  on for  a  cataract and  46  had  ocular changes that consisted
                                     11-40

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mainly of lens opacities and/or nuclear sclerosis.  To eliminate the influence  of
age on the development  of the lens changes, the painters were compared with age-
matched  unexposed  railroad  engineers; 69  age^matched  pairs  were generated for
comparison.  Results showed that, in 27 instances, more lens changes were present
in the car painters than in the  age-matched  engineers,  and that  in  4  instances,
there were more changes in the engineers  (Table  11-13).
                                   TABLE  11-13
           Frequency of Lens Changes  and  Distribution  by  Exposure Time
  I        in 69 Age-Matched  Pairs of Car Painters and Railway Engineers3
Pesult

Car painters had fewer
changes than the engineers
No noticeable difference
between the pairs

Car painters had more
chknges than the engineers
Frequency of
Lens Changes
(no. pairs)

4

38


27
Distribution
by Years
£ 10 11

3

22


6
of Lens Changes
of Exposure
to 20 >21

1 0

13 3
i

17 4
Source:   Raitta et  al.,  1976
 In the remaining  38  pairs,  both  the  painters  and the unexposed engineers  had
 similar lens  changes.    The lens  changes  were further  found to  occur with
 increased  frequency after 10 years of  exposure  (Table 11-13).
 11.7.2. Sensory Thresholds. Gusev (1965)  investigated the  olfactory  threshold
 for toluene in  30   subjects  with  a  total of  744  observations.   The  minimum
 perceptible  concentration was found by  this Russian investigator to  be  within
 0.40 to 0.85 ppm (1.5 to 3.2 mg/m  ), and the maximum imperceptible concentration
 within 0.35  to 0.74 ppm (1.3 to  2.8 mg/m3).   In sniff tests  with 16 subjects (8
 male,  8 females), May (1966) determined the minimum perceptible concentration to
 be a much  higher 37 ppm (140 mg/m-*); toluene was found to be  clearly perceptible
 at 70  ppm. In the latter study,  the number of observations used to establish  the
 average values were not stated.
      Odor  thresholds  and  sensory responses to inhaled vapors of "toluene concen-
 trate"  were more  recently  determined  by Carpenter  et al.  (1976).    "Toluene
 concentrate"  is a hydrocarbon mixture containing 15.89? toluene, 38.69?  paraf-
 fins,  15.36? naphthenes,  and 0.06?  benzene.  The most probable  concentration  for
 odor threshold,  determined in two  trials  with 6 subjects, was 2.5 ppm.   Based on

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sensory thresholds for  irritation (eye, nose,  throat),  dizziness,  taste,  and
olfactory fatigue, 6  of 6 volunteers  indicated their willingness  to work for
8 hours in a  concentration of 480 ppm  (corresponding to 220  ppm  of toluene).
Only 3  subjects  thought  they  could work  in an atmosphere  containing 930 ppm
(corresponding to about  427 ppm toluene).
11.8.  EFFECTS ON THE  SKIN
     Toluene  appears  to be absorbed less readily  through the skin than through
the  respiratory  tract,  but percutaneous  absorption  of  liquid toluene  may be
significant (Section  13.1.).  When toluene is applied to the skin, its degreasing
action  will  remove natural  lipids,  possibly  causing  dryness,  fissures,  and
contact dermatitis (Gerarde,  I960; Browning, 1965).
     Malten et al.  (1968) found that exposure of  human  forearm skin for  1 hour on
6  successive  days to toluene  (volume  and  conditions not stated)  resulted in
injury  to  the epidermal stratum  corneum  (horny layer).   The skin  damage was
assayed by measurements  of water vapor loss, and  daily  measurements following the
exposures indicated that regeneration took about 4 weeks.
     Koilonychia and  hapalonychia of  the fingernails  (conditions  in which the
nails are, respectively, concave and uncornif.ied  (soft)) were  observed in 6 of 16
cabinet makers who were  dermally exposed to a thinner  mixture that contained 30J
toluene, 30?  xylene, and 40? methyl alcohol  (Ancona-Alayon, 1975).  These defor-
mities  involved  primarily the thumb,  index,  and  middle fingernails,  and were
attributed to the  practice of  cleaning metal parts on  furniture  with solvent-
soaked rags and unprotected hands.  Most of the  affected workers had an average
exposure of 2 years.
11.9.  SUMMARY
     Toxicity studies in  humans have primarily involved evaluation of indivi-
duals exposed to  toluene via inhalation  in experimental or occupational settings
or  during episodes of  intentional abuse,  and  the  health effect  of greatest
concern is dysfunction of  the CNS.
     Single 8-hour experimental  (Von Oettingen  et  al.,  1942a,  1942b; Carpenter
et al., 1944) and subchronic occupational (Wilson,  1943)  exposures to toluene in
the  range of  200  to 300  ppm have elicited subjective symptoms indicative of CNS
toxicity  (e.g.,  fatigue,  nausea,  muscular weakness,   mental  confusion,  and
impaired  coordination).   These  effects   were  generally  dose-dependent  and
increased in  severity  with increasing toluene concentration.  Acute-experimental
exposures to  to?uene have  also  caused objective increases in reaction time at 200
                                     11-42

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                                 I
to 300  ppm (Ogata et  al,,  1970; Gamberale and Hultengren,  1972) and decreases in
perceptual speed  at  700  ppm  (Gamberale and  Hultengren,  1972).   Gusev (1965)
observed disturbances of  EEC  activity in several  subjects exposed to 0.27 ppm
toluene for  6-minute intervals,  but  this  effect  does  not  have  any apparent
toxicological significance.
     Short-term  accident;,! -workplace  (Lurie,  1949;  Browning,   1965;  Longley
et a!,, 1967; Reisin  et  al.,  1975) and deliberate  (Press  and Done,  1967a, 1967b;
Wyse,  1973; Lewis and Patterson,  1974;  Hayden  et al., 1977;  Oliver and Watson,
1977; Barnes, 1979; Helliwell and Murphy,  1979) inhalation exposures to exces-
sive levels  of  toluene  (i.e., levels approaching air saturation concentrations
of  30,000 ppm)  have  initially   resulted  in  CNS  stimulatory effects  such  as
exhilaration, lightheadedness,.dizziness,, and delusions.   As  exposure durations
increase,   narcotic  effects   characteristic  of  CFS  depression   progressively
develop, and, in extreme cases, collapse, loss cf consciousness, and death (Winek
et al., 1968; Chiba,  1969; Nomiyama and Nomiyama,  1978)  have  occurred.
     Chronic occupational exposure to toluene has  been associated  with "nervous
hyperexcitability" (Parmeggiani  and  Sassi,  1954)  and subjective memory, think-
ing, and  activity disturbances   (Munchinger,  1963)  in workers exposed, respec-
tively, to concentrations of 200 to 800 ppm and 300 to 430 ppm.  No evidence of
adverse neurological  effects  have been reported,  however, in other studies of
printers exposed to 200  to 400 ppm toluene  (Suhr,  1975) or  manufacturing workers
exposed to 80 to  160 ppm toluene (Capellini and  Alessio, 1971),  although the
negative  findings  in the  former study are  equivocal and  symptoms  of stupor,
nervousness, and insomnia were noted  in one worker exposed to higher concentra-
tions  (210 to 300 ppm) of toluene in  the latter study.   Exposure to mixtures of
vapors  from  an organic solvent  containing  predominately low-levels of toluene
 (approximately 30 ppm) for an average of  15  years has produced a  greater inci-
dence  of  CNS symptoms  and  impaired  performance  on  tests for intellectual and
psychomotor  ability  and  memory  in  car   painters   (Hannir.en  et al.,  1976;
Seppalainen  et  al.,   1978).   Matsushita et  al.  (1975)  reported impaired per-
formance in neurological and muscular function tests in female shoemakers who had
been exposed  to  15 to 200 toluene for an average  duration of over 3 years, but
these  workers were also exposed  to "slight" levels of gasoline.  Changes in EEC
response  to photic  stimulation  were reported  by  Rouskova  (1975)  in workers
exposed to >250 ppm toluene and  unspecified levels of 1,1,1-trichloroethane for
an average of 13.5 years.
                                      11-43

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     Residual  effects  indicative  of  cerebellar  and  cerebral  dysfunction have
been observed  in  a number of persons who had abused toluene or solvent mixtures
containing toluene over  a period of  years (firabski,  1961;  Satran and Dodson,
1963; Knox and Nelson, 1966; Kelly,  1975;  Boor and Hurtig, 1977; Weisenberger,
1977; Keane,  1978; Sasa et al.,  1978;  Tarsh,  1979;  Malm and Lying-Tunell,  1980).
These effects  were largely reversible upon cessation of exposure, but prolonged
toluene abuse  has, on occasion, led to permanent encephaiopathy and brain atrophy
(Kno* and Nelson, 1966; Boor and Hurtig,  1977; Sasa  et  al.,  1978).  Reports of
     \
polyn'europathies   in  abusers of glues and  solvents  have  also  appeared  in the
literature, but have in all cases involved mixtures  of  toluene  and  other solvents
such  as  n-hexane  and  methyl ethyl  ketone  (Matsunmra  et al.,  1972;  Takenaka
et al.,  1972;  Goto et al.,  1974;  Shirabe et al.,  1974;  Suzuki  et  al.,   1974;
Korcbkin  et al;,   1975;  Oh and Kim,  1976; Towfighi  et al.,  1976;  Altenkirch
et al.,  1977).
     Early reports of  occupational  exposures (generally  prior  to  the   1950s)
ascribed myclotoxic effects to toluene (Greenburg et al.  1942;  Wilson, 1943), but
the  majority of recent evidence indicates tnat 'toluene is  not  toxic to the  blood
       i
or bone marrow (Von Oettingen  et al.,  1942a, 1942b; Parmeggiani and Sassi,  1954;
Banfer,  1961;  Capellini and Alessio,  1971: Suhr,  1975; Matsushita et al.,  1975).
When administered orally to leukemia patients, it has  been  further reported that
toluene  was nontoxic  and  had no  effect  on the  leukemic  process (Francone and
Braier,  1954).   Hematological abnormalities  have been infrequently reported in
sniffers of toluene-based glues and thinners (Christiansson and Karlsson,  1957;
Msssengal* et  al., 19&3; Sokol and Robinson, 1963;  Barman  et al.,  -1964; Press and
Done,  1967b),    Other  investigators have  noted  increases in prothrombin time
 (Pacseri  and  Emszt,  1970),  decreases   in  phagocytic  activity  of  leukocytes
 (Bansagi,  1966),   and  increased   enzyme  concentrations  in  leukocytes  and
lymphocytes (Fribor-ska, 1973) of workers who were exposed to toluene.  Decreases
 In  serum icmunoglobin and complement levels (Lange et al.,  1973a;  Smolik et al.,
 1973)  and  leukocyte agglutinins  (Lange  et al.,   1973b)  have  been  reported in
workers  exposed simultaneously to benzene, toluene, and xylene.
     Liver enlargement was reported in an early study  of painters  with exposures
 to  100 to  1100 ppm toluene for  2  weeks  to more than  5 years  (Greenburg et al.,
 1942), but this effect was not associated with clinical or laboratory evidence of
 disease or corroborated in subsequent studies  of  workers  (Parmeggiani and  Sassi,
 1954; Suhr, 1975).  Chronic occupational exposure to toluene has generally not
                                     11-44

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been   associated   with  abnormal  liver  function   (Greenburg  et  al.,  19^2}
Parmeggiani and Sassi,  1954; Capellini and Alessio,  1971; Suhr ,  1975; Kurppa and
Husman,  1982),  although reductions in serum  bilirubin  and  alkaline phosphatase
(Szadkowski  et  al.,  1976)  and  increases   in  gamma  glutamyl  tranapeptidase
(Trevisan and Chiesura, 1978) have been noted.  Intensive exposure to toluene via
glue  or thinner  sniffing appears  to have a minimal  effect  on liver function
indices  (Christiansson and  Karlsson,  1957;   Grabski,  1961;  Massengale  et al.,
1963;  Sokol and Robinson, 1963; Barman et al.,  1964; Boor and Hurtig,  1977; Press
and Done,  1967a,  1967b).
     Exposure  to  mean concentrations of  100  tc 1100 ppm toluene for 2 weeks to
5 years  (Greenburg  et  al., 1942)  or 60  to   100 ppm toluene  for  over  3 years
(Matsushita  et al.,  1975)   did  not result  in abnormal  urinalysis findings in
airplane painters or female shoemakers, respectively. Glomeruiar filtration rate
was  reduced  in  rotogravure  worker."  with  uncharactsrized  toluene  exposures
(Asker'gren et  al., 1981), but clinical case  reports have described proteinuria
and hematuria (Lurie,  1949; O'Brien  et  al.,  1971)  and myoglobenuria and renal
failure (Reisin et al.,  1975)  in workers who  were  accidentally overexposed to
toluene.   Pyria,  hematuria,  and  proteinuria  have been  the  most  frequently
observed signs  of  renal dysfunction associated with  the deliberate  inhalation of
toluene-based  glues,  but  these effects have not been universally observed in glue
sniffers (Christiansson and Karlsson, 1957;  Massengale et  al., 1963; Sokol and
Robinson,  1963; Barman  et  al.,  1964;  Press  and Done,  1967a, 1967b).   Several
reports have recently appeared that  associate deliberate inhalation of toluene
with metabolic  acidosis (Taher  et al., 1974; Fischman and Oster, 1979a; Koeger et
al., 1980; Bennett and Forman,  I960;  Moss et al., 1980).
     Acute experimental  exposure to  toluene  within the  range of 50 to 800 ppm
have  not   caused  any  definite  effects  on  heart  rate  or  blooci  pressure
(Von Oettingen et al., 1942a,  1942b; Ogata et al..  1970; Astrand et al., 1972;
Gamberale  and  Hultengren, 1972).  Toluene has  been implicated in a small  number
of sudden deaths  due to solvent sniffing which appear  to  result from cardiac
arrhythmias  (Bass,  1970;  Alha et  al.,  1973),  but  trichloroethane and fluorinated
aerosol propellants have most  frequently been  associated with these deaths.
     Subjective complaints  of dysmenorrhea have  been  reported by a significant
number of  female  shoemakers  exposed to 60 to 100 ppm toluene  and concomitantly to
20 to 5C  ppm  gasoline in  a "few" working  places  for an  average duration of
3 years and 4 months  (Matsushita et al., 1975).   Disturbances of menstruation
                                     11-45

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have  also  been  reported in women exposed concurrently  to toluene, benzene, and
xylene  in  the workplace (Michon, 1965), and  in  women exposed occupationally tp
toluene and  other  unspecified solvents (Syrovadko, 1977), but a specific effect
of toluene could  hot  be determined  from the available data.  Information on the
possible reproductive effects of toluene in males is not available.
     Minimum perceptible  concentrations  of toluene have  been  determined to be
0.^0  to 0.85 ppn  (Gusev,  1965) and  37 ppm (May,  1966),  but the reasons for this
discrepancy are not apparent.  Toluene has  been  reported  to cause transitory eye
and respiratory tract irritation as a  result of 8-hour exposures in the range of
200 to 800 ppm (Carpenter et  al.,  19^; Partneggiani  and 5ass.i, 195^; Capellini
and  Alessio,  1971),   but  no  complaints  of  respiratory  tract  discomfort  were
recorded in volunteers or workers exposed  to  levels  as high as 600 to 1500 ppm
for  8-hour  periods in other  studies  (Von OettiP^cn  et al.,  19^2a,b;  Wilson,
19^3)-   No complaints of respiratory tract or eye irritation were recorded in men
accidentally exposed to  10,000  to  30,000  ppm  toluene  for   brief  durations
(Longley et al.,  1967).
     Transient  epithelial injury  to  the eyes  that healed within  ^8 hours was
observed in workers who were  accidently splashed with toluene (McLaughlin, 19^6;
Grant,  1962).  Opthalmologic examinations of spray painters who were exposed to
100 to 1000 ppm toluene for  2 weeks to more than 5 years were normal (Greenburg
et al., 13^2), but Raitta et al.  (1976) found  lens  chenges ir. a  group  of car
painters exposed   concurrently to  approximately  30 ppm toluene and  much lower
concentrations  of  other solvents for an average of 15  years.  The little informa-
tion that  is available on  the  dermal toxicity of toluene indicates that moderate
contact may cause skin  damage due  to its  degreasing action  (Gerarde, 1960;
Browning,  1965; Malten et  al., 1966).

11.10  REFERENCES

ALHA,  A.,  KORTE,   T. and TEAHU,  M.    (1973).    Solvent  sniffing death.   Z.
gechtsmed.  72: 299-305.

ALTENKIRCH,  J., MAGER, J.,  STOLTENBURG,  G. and HELKBRECHT,  J.   (1977).   Toxic
polyneuropathies  after sniiTing a  glue thinner.   J. .Neurol.  21*4(2): 157-152.
                                     11-146

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ANCONA-ALAYoN',  A.    (1975).   Occupational  koilonyChia  from  organic  solvents.
Contact Dermatitis.  J^: 367-369.

ASKERGREN, A.,  BRANDT,  R.,  GULLQUIST, R., SILK,  B. ,  and STRANDELL. T.  (19B1).
Studies on kidney function  ir, subjects  exposed to organic solvents.  Acta  Med.
Scar.d. 210(5):  373-376.
  ,      S

ASTRAND,  I.,  EHRNER-SAMUEL,  H. ,  KILBOM,  A.   and OVRUM, P.   (1972).   Toluene
exposure. I. Concentration in alveolar air and  blood at rest and during exercise.
Work Environ. Health.   72 ( 3_) :  119-130.

BANKER, W.  (196l).  (Studies on  the  effect of pure toluene on the  blood picture
of  photogravure   printers   and  helper   workers.]     Zentralbl.   Arbeitsmed.
VU  35-^0.  (In Ger.).  (Citec  in NIOSH,  1973).
         i
BANSAGI,  J.   (1968).   [Effect  of toluene on  the phagocytic  activity  cf white
blooc! cells in  printers.]   Kunkavedelea.   14;  26-28.   (In Hungarian; summarized
in Chea. Abstr. 69:895^^3,  1966).

BARMAN, M.L.,  SIEGEL ,  N.B., BEEDLE,  D.B. and  LARSON, R.K.   (1964).  Acute and
chronic effects of glue sniffing.  Calif. Med.  100:  19-22.

BARNES, G.E.   (.1979).   Solvent  abuse:  A  review.  Int. ^J. Addict.  _ljf: 1-26.

BASS, K.   (1970).  Sudden sniffing  death.  J_.  Aner.:Med. Assoc.   212:  2075.

BENNETT,  R,H.  and  FORMAN,   H.R.    (1980).   Hypokalemic periodic  paralysis in
chronic toluene exposure.   Archives £f  Neurology.  37 ( 10):  673.

BOOR, j.w. and HURTIG,  H.I.   (1977).  Persistent cerebellar ataxia after exposure
to toluene.  Ann Neurol.
BROWNING, E.  (1965).  Toxicity and Metabolism of Industrial Solvents.  New York:
Elsevier Publishing  Co., p. 66-76.

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CAPELLINI,  A.  and ALESSIO,  L,   (1971).   [The urinary excretion of hippuric acid
in workers  exposed to toluene.]  Med.  Lavoro.  62: 196-201.  (In Ital.).

GASPENTER,  C.P.,  SHAFFER, C.B., WEIL, C.S. and SMYTH,  H.F.,  JR.  (194*0.  Studies
on the inhalation of  1,3-butadiene; with a comparison of its narcotic effect with
benzol, toluol,  and  styrene,  and a note on-the  elimination of styrene  by the
human.  J.  Ind. K_yg_.  Toxicol.   26_:  69-»78.

CARPENTER.  C.P.  et  al. (1976).  Petroleum  hydrocarbon toxicity studies. XIII.
Anioal ana  human response  to vapors of  toluene concentrate.   Toxicol. Appl.
Pharmacol.   36: 473-490.

CHIBA,  P..     (1969).    Sudaen  death from   thinner.    Nichidai  Igaku  Zasshi.
26_: 952-998.  Taken from:   Chem. Abst.   72:64867g, 1969.

CHRISTIANSSON, G. and KARLSSOK,  B.   (1957).   "Sniffing1 - berusningssatt bland
barn.  Svensk  Lakartidn. 5^:  33.   (In Swedish; cited in Press and Done,  1967b).

CIESLINSKA, A.,  KOWAL-GIERC2AK,  B, , KUCZYNSKA-SEKIETA,  K.,  MALOLEPSZY, J. and
WRZYSZCZ, M.   (1969)*   [Serum iron and  copper levels  in subjects  with chronic
toluene exposure.]   Pol. Tyg.  L_ek.   2±:  1848-1850.   (In Pol.).

PRAGOSICS,  B., FERENCI,  P., PESENDORFER, F.  and  WEWALKA,  F.G.   (1976)   Gamma-
glutamyltranspeptidase  (GGTP): Its  relationship  to other enzymes for diagnosis
of liver disease.  Progress in Liver Disease.  5_: 435-449.

FAILLACE,  L.A.  and GUYNN,   R.W.   (1976).   Abus  of  organic  solvents.   Psycho-
somatics.   17(4): 188-189.

FERGUSON,  T.,  HARVEY,  W.F. and  HAMILTON,  T.D.   (1933).   An  inquiry into the
relative toxicity of benzene and toluene.  I. Hyg.   3J3: 547-575.

FISCHMAN, C.M. and OSTER, J.R.  (1979).  Toxic effects of toluene.  A new  cause of
high anion gap metabolic acidosis.   J. Am. Med. Assoc.   241(16):  1713-1715.

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FRANf'ONE,  M.P. and  BRAIER,  L.   (195^).   [The  basis  for the  substitution of
benzene by the higher  homologues  in industry.]  Med.  Lavoro.   ^5: 29-32.   (In
Ital.).

FRIBORSKA, A.   (1973).  Some cytochemical  findings  in the  peripheral white blood
cells in  workers  exposed to toluene.   Folia  Haematol.    (Leipzig).   9_9 : 233.
(Cited in NRC, 1980).

GAMBERALE, F.  and I^TENGREN,  M.  (1972).  Toluene exposure. II,   Psychophysio-
logical functions.  Work Environ.  Health.  9(3): 131-139.

GELLMAN, V.   (1968).   Glue sniffing ajnong Winnipeg school children.  C?n. Ked.
Assoc. J.  98: ^1
GERARDE, H.W.   (I960).   Toxicology and , Biochemistry of Aromatic Hydrocarbons.
New York:  Elsevier Publishing Co., p. 141-150.

GOTO, I., MATSUMURA, K. and INOUE, N.   ( 197M ).   Toxic polyneuropathy due to glue
sniffing.  ^J. Neurol. Neurosurg. Psychiat.  37(7) : 84b-853-

GRABSKI, D.A_.    (1961).   Toluene  sniffing producing cerebellar degeneration.
Amer. J[. Psychiatry.  1 16:
 GRANT, W.M.  (1962).  Toxicology of the eye.  Springfield,  IL , Charles C. Thomas,
 p. 5^-545.  (Cited in NIOSH, 1973).

 GREENBURG, L.,  MAYERS,  M.R., HEIMANN, H. and MOSKOWITZ,  S.  (19^2).  The effects
 of exposure to toluene in industry.  ^J. Amer. Hed. Assoc.  118: 573-578.

 GUSEV, I.s.   (1965).   Reflective  effects  of microconcentrations  of benzene,
 toluene,  xylene  and  their comparative  assessment.   Hyg.  Sanit.   30: 331-335.
 (Russian report published in English).

 HANNINEN, H., ESKELINEN,  L.,  HUSMAN, K. and  NURMINEN,  M.  (1976).  Behavioral
 effects of long-term  exposure to a  mixture of  organic solvents.  Sctind. J,  Work
 Environ.  Health.   2(*0: 2^0-255.
                                     11-149

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HAYDEN,  J.W.,  PETERSON,  R.G.  and BRUCKNER,  J.V.  (1977).  Toxicology of toluene
(methyl benrene):  Review  of current literature.  Clin. Toxicol .   11 ( 5 ) : 5^9-559.

KELLIWELL,  M.  and  MURPHY,  M.    (1979).   Drug-induced  neurological  disease
(letter).   Brit.  Med.  J.   1(6l7j): 1283-1261.

KEAKE,  J'.R.   (1978).  Toluens optic neuropathy.  Ann. Neurol .  4(4): 390.

KELLY,  T.W.   (1975).   Prolonged  cerebellar dysfunction  associated  with paint
sniffing.   Pediatrics.  5_6: 605-606.

KNOX, J.W. and  KELSON,  J.R.    (1966).   Penaanent  encephalopathy from toluene
inhalation.  N.  Eng. J_.  Med.   275: 1494-1196.

KOROBKIN,-  R.  et al .    (1975).    Glue  sniffing  neuropathy.     Arch.  Neurol .
i_2: 15&-162.

KOWAL-GIERCZAK,   £.,  KUCZYNSKA-SEKIETA ,  K . ,  CIESLINSKA,  A.,  WRZYSZCZ,  H.  and
KALOLEPSZY, J.   (1969).    [Some biochemical  tests in  subjects   occupationally
exposed tc toluene.]  Pol. Tj£.  Lek.  2^: 1662-1685.  (In Pol.).

KP-GEGER, R.M., MOCRE,  R.J. and LEHMAN, T.H.   (1980),  Recurrent  urinary calculi
associates with  toluene  sniffing.  ,;. Urol .   123( 1) : 89-91.

KURPPA, K., and  HUSMAN,  K.   (1962).   Car  painters  exposure  to a  mixture of
organic solvents.  Serum  activities  of liver  enzymes.  Scand. J_. Work Environ .
 LANGE, A. et al .  (1973a).   Serum  immunoglobulih levels  in  workers exposed  to
 benzene, toluene,  and xylene.   Inter. Arch, fuer Arbeilsmedizxa.  3K 1): 37-41.
       A. et al .   (1973b).  Leukocyte  agglutinins  in workers  exposed  to  benzene,
 '.oluene and xylene.   Int.  Arch.  Arbeitsmed.  31: 45-50.
                                     11-50

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LEWIS,  P.W., PATTERSON, D.W.  (197*0.  Acute and chromic  effects of the voluntary
inhalation  of  certain commercial volatile solvents by juveniles.  J.  Drug  Issues.
1(2);  162-175.

LINDER, R.L.,  LERNER,  S.E.  and WESSON,  D.R.   P975).   Solvent  sniffing:  A
continuing  problem among youth.  Proc. West pharmaccl. Soc.  18: 371-37«.

LINDSTROEM, K.  (1973).   Psychological performance of workers exposed to  various
solvent.  Work Environ.   10(3): 151-155.

LITT,  I.F., COHEN, M.I., SCHCNBERG,  S.K. and SPIGLAND,  I.  1972.  Liver  disease
in the drug-using adolescent.  J.  Pediatr.  in :  23&-212.

LONGLEY, E.G., JONES,  A.T., WELCH, R. and LOMAEV.  0.   (1967).   Two acute  toluene
episodes in merchant ships.  Arch. Environ. Health.  1*4: 161-167.

LURIE, J.B.   (1919).  Acute toluene poisoning.  S. Africa Hed. J_.   23: 233-236.

MALM, G. and LYING-TUNELL, U.  (19faC).  Cerebellar dysfunction related to toluene

KALTEN, K.E.,  SPRUIT, D.  and DEKEIZER, M.J.M.   (1966).  Horny layer injury  by
solvents.  Eeruf sdercatosen.  16:  135-117.

USSENGALE, 0.,%.,  GLASER,  H.H.,  LELIEVRE,  R.E.,  DODDS, J.B.  and KLOCK, M.E.
 (1963).  Physical and psychologic factors  in glue sniffing.   N.  En£l.  J_. Hed.
2_69: 13140-1314**.

MATSUKURA,  M., SNOVE, N.  and  OHNISKI, A.   (1972).   Toxic polyneuropathy  due  to
glue sniffing.  Clin.  Neurol.  \2_: 290-296.

MATSUSHITA, T.  et al.   (1975).    Hematological  and  neuro-ouscular  response  cf
workers exposed to low concentration  of toluene vapor,  ^nd. Health,  jj;  115.

MAY, J.  (1966).  Odor  thresholds  of solvents for  assessment  of solvent odors  in
the air.  Straub.  26(9): 31-38.
                                     11-51

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MCLAUGHLIN,  R.S.    (19^6).   Chemical  burns of  the human  cornea.    Amer.  Jf.
Ophthalsol.  29: 1355-1362.

M^TRICK,  S.A., BRENNER  R.  P.   (1962).    Abnormal  brainstem  auditory evoked
potentials in  chronic paint  sniffers.   Ann.  Neural.  12: 553-556.

MICHON, S.   (1965).  [Disturbance  of menstruation in wcaen working in  an atmos-
phere  polluted with aromatic hydrocarbons].  Pol. Tyg. Lek.  20: 15^7-16^9.   (In
Polish with  English summary).

KOSS,  A.M.,  GABOW, P.A.,  KAEHNY,  W.D.,  GOODMAN,  S.I. and  HAUT,  L.L.   (1960).
Fancor.i's  syndrose and distal  renal  tubular  acidosis after glue sniffing.  Anr.,
Interr.. Mec.   92.: 69-70.

MUNCKINGER,  R.  (1963).  Der nachweis central nervoser storungen bei losungsiiitt
elexpor.ierter,  Arbeitern.  Excepta Medica Series, Madrid; 16-21.  2(62):  687-669.

KIOSH (NATIONAL INSTITUTE  FOR OCCUPATIONAL SAFETY AND HEALTH).  (1973).  Criteria
for a RecotsEenaec1  Standard.   Occupational  Exposure to Toluene.   Final Report.
Contract  No.  HSM-99-72-116.   Available  through N7IS,  N7IS  No. PB-22-219/8,
108 p.

NOMnAMA.  K. anc  NOKIi'AMA, H.   (197&).   Three  fatal cases of thinner-sniff ing,
anc experimental exposure  to toluene in humans  and  aninais.   Int.. Irch. Occup.
rnvircri.  Health.   j.1( 1): 5^-B^.

NSC (NATIONAL RESEARCH COUNCIL).    (I960).   The Alkyl Benzenes.   Committee en
Alkyl Benzene  Derivatives, Board on Toxicology and Environmental Health "azartis;
Assembly  of  Life Sciences, National  Research Council.  Washington, DC:  National
Academy Press.

O'BRIEN,  E,T.,  YEOMAN, W.D.  and  HOBBY,  J.A.E.   (1971).  Hepatorenal damage from
toluene in a "glue sniffer."  Brit.  Me^l. J.   2: 29-30.
                                     11-52

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OGATA, M., TOMOKUNI, K.  and TAKATSUKA,  Y.  (1970).  Urinary excretion  of hippuric
acid and m- or p-methylhippuric acid in the urine of persons exposed to vapors  of
toluene  and  m-  or p-xylene  as  a  test  of  exposure.    Brit.  J.   Ind.  Med.
27(1): ^3-50.
        \
OK,  S.J. and  KIM, J.M.  (1976).  Giant axonal swelling in "buffer's"  neuropathy.
Arch.  Neurol.  33(8); 583-586.

OLIVES, J.S.  and WATSON,  J.M.   (1977). Abuse of solvents "for kicks": A review
of 50  cases,  l.ancet.   1(8002):  8486.

FACSEKI,  I.  and  EMSZT,  G.   (1970).  Medical  aspects of the exposure to toluol.
Kunkavedelets.  J_6_:  41-J*6.  (In  Hungarian; cited in NIOSH, 1973).
          i
PARXEGGIANI,  L.  and SASSI, C.   (1954). [Occupational  risk of  toluene: Environ^
cental studies  and clinical  investigations of chronic  intoxication].   Med.
Lavoro.   V>:  574-63.   (In Ital.).

POHL,  K.  and  SCKMILDE,  T.  (1973).   [Serum  concentration and performance changes
following  repeated   inhalation   of   eleven   technical   organic   solvents.]
fal-jtalkohol.  J_G: 95-120.  (InGer.J.

POWARS,  D.  (1965).  Aplastic anemia  secondary to glue sniffing.  _N. Engl.   _J.
Med.  2?;: 700-702.

PRESS, E.  ar.d .DONE, A.K.   (1967a).   Solvent  sniffing.  Physiologic  effects and
community control measures for intoxication from the  intentional  inhi _ation  of
organic  solvents. I.  Pediatrics.   _39:  451.

PRESS, E.  and DONE, A.K.   (19675).  Solvent sniffing.   Physiologic  effects and
community control  measures for intoxication from the  intentional  inhalation  of
organic  solvents. II.  Pediatrics.   _3_£: 611.

RAITTA,  C.,  HUSMANN,  K.   and  TOSSAVAINEN,  A.    (1976).   Lens  changes  in car
painters  exposed to a  mixture  of  organic  solvents.   Albrecht \r. Grafes  Arch.
          200(2): 149-156.
                                     11-53

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REISJN, E.,  TEICHER,  A.,  JAFFE,  R. and ELIAHOU,  H.E.  (1975).  Myoglobi.iuria and
renal failure in toluene poisoning.   Brit.  J_. Indust.  Med.  32(2): 163-164.

ROUSKOVA, V.  (1975).  Photic stimulation in early  diagnosis  of the effects of
some  h'armful  industrial  substances on  the  central  nervous  sytem.  Int.  Arch.
Arbeitsmed.  3':(4): 283-299.

SASA,  M.(  IGARASHI,  S. ,  MIYAZAKI,  T., MIYAZAKI,  K.  and  NAKANO, S.   (1978).
Equilibrium disorders with diffuse  brain  atrophy in long-term toluene sniffing.
Arch.  Otorhinolarngol.  22U3):  163-169.

SATRAN,  R. and  DODSON,  V.   (1963).   Toluene habitation - report  of a case.  _N.
En£.  J.  Med.  263(13): 219-220.
_.       _        .

SEPPALAINEN, A.M., HUSMANN,  K.,  and MARTENSON,  C.   (1978).  Neurophysiological
effects  ^f long-term exposure to a mixture of organic solvents.  Scand. J_.  Work
Environ. Health.   4(4): 304-314.  Taken from:   Chem. Abst.  9C):156383w, 1979.

SHIRABE, T., TSUDA, T., TERAO,  A. and ARAKI, S.   (1974).   Toxic  polyneuropathy
due to glue-sniffing:'  Report of  two  cases  with a light and electronmicroscopic
study of the peripheral nerves and  muscles.  J^. Neurol. ScjL.  21(1): 101-113.

SlrlOLIK,  R. et al.  (19-73).  Serum complement level in workers exposed to benzene,
toluene  and xylene.   Int. Arch. Arbeitsmed.  ^J_: 243-247.

SOKOL, J. and ROBINSON, J.L.  (1963).  Glue sniffing. Western Medicine.  j|4:  192.

 >UHR,  E.   (1975).  Comparative  Investigation of  the State of  Health of Gravure
 'rinters Exposed  to  Toluene.  Gesellschaft zur  Forderung des Tiefdrucks E.V.,
 'eisbaden, Federal Republic  of Germany.  92 p.

  UZUKI,  T., SHIMEO, S. and NISHITANI, H.   (1974).  Muscular atrophy due to glue
          Int. Arch. Arbeitsmed.   33(2):  115-123-
                                     11-54

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SYROVADKO,  O.N.   (1977).  Working conditions and health status of women handling
organosiliceous  varnishes containing toluene.  Gig,  Tr. Prof, Zabol,  12: 15-19.
(In Russian with English summary).

SZADKOWSKI, D.,  PETT,  R., ANGERER, J., MANZ, A. andLEHNERT, G.   (1973).  Chronic
solvent exposure at work,  II.  Harmful  material  levels in  blood  and excretion
rates of metabolites in urine with the importance of environmental criteria for
toluene exposed  printers.  Int. Arch. Arbeitsmed.  31(4): 265-276.
     \ -

SZADKOWSKI, D. et al.   (1976).   Evaluation of occupational exposure to toluene.
Medizinische Monatsschrift.  ?0( 1):

TAHER, S.M.,  ANDERSON,  R.J., MCCARTNEY,  R;  POPVITZER, M.M. and  SCHRIER,  R.W.
(1974).  Renal tubular acidosis associated with toluene sniffing.  N. Engl.  _J.
Med.  290: 765-768.

TAKENAKA, S., TAWARA,  S.,  IDETA, T.,  OKAJIMA, T. and TOKUOMI, H.  (1972).  A case
      i
with polyneuropathy due to glue-sniffing.  Clin. Neurol.  12: 747.

TARSH, M.J.  (1979).  Schizophreniform psychosis  caused by sniffing toluene.  _J.
Soc. Occup. Med.  29(4): 131-133-

TOWFIGHI, J., GONATAS, N.K., PLEASURE, D., COOPER, H.S. and MCCREE, L.  (1976).
Glue sniffer's neuropathy.  Neurology.  26: 238-243.

TREVISAN, A. and CHIESURA, P.  (1978).  Clinical research on the hepatotoxicity
of  toluene.  Ital. J_.  Gastroenterology.   10(3): 210.

VON OETTINGEN,  W.F.,  NEAL,  P.A. and DONAHUE, D.D.   (1942a).   The toxicity and
potential  dangers  of  toluene—Preliminary report.    J_,  Amer.  Med.  Assoc.
118: 579-584.
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VON   OETTINGEN,    W.F.,    NEAL,   P.A.,   DONAHUE,   D.D.,   SVIRBELY,   J.L.,
BAERNSTEIN,  H.D.,  MONACO, A.R., VALAER, P.J. and  MITCHELL,  J.L.   (1942b).  The
Toxicity and Potential Dangers of Toluene, with Special Reference to its Maximal
Permissible Concentration.  U.S. Public Health Serv. Pub. Health Bull. No. 279,
50 p.

WALTER,  P.V.,  MASLYN, R.T.,  SHAFFER,  G.P.  and  DANIELS,  C.A.   (1977).   Glue
sniffing:  The continuing menace.  Drug Forum.  5(3): 193-197.

WATSON,  J.M.   (1979).    Glue  sniffing.    Two   case  reports.   Practitioner.
222(1332): 81)5-8*47.

WEISENBERGER,  B.L.      (1977).     Toluene   habituation.     _J.   Occup.   Med.
19(8): 569-570.

WILSON,  R.H.   (19*13).  Toluene poisoning.  _J. Amer. Med. Assoc.  12j: 1106.

WINEK, C.L., WECHT, C.H. and  COLLOM, W.D.   (1968).   Toluene fatality from glue
sniffing.  Penn. Med.  JJ_: 81.

WINNEKE, G., KASTKA, J. and FODOR, G.G. (1976).   Psychophysiological Effects of
Low Level Exposure to Trichloroethylens and Toluene.  In:  Proceedings of the 2nd
International   Industrial   and   Environmental   Neurology   Congress,   Prague,
Czechasiovakia, 197**, (Klimkova-Deutschova,  E.  and Lukas, E., eds.).  Univerzita
Karlova  Praha, p. 78.

WYSE,  D.G.   (1973).  Deliberate  inhalation of volatile hydrocarbons:  A review.
Can. Med. Assoc. J.  108: 71-71.
                                      11-56

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                            12.  ANIMAL TOXICOLOGY

12.1.   SPECIES SENSITIVITY
     information on  the toxic  effects'  of  chronic exposure  to  low levels  of
toluene may be more relevant to  greater  numbers  of people than  information  on
acute toxicity (see Sections 10.3 and 10.f).  However,  for those  rare exposures
to high  levels   of toluene,  data obtained from  acute  toxicity  studies  are
valuable.  In the  sections  to follow, consideration will  be  given  to  acute  as
well  as chronic  studies.
     Inhalation  has been the principal route  of  exposure  in  humans;  therefore,
animal studies  have  centered  on intoxication  by this  route.   In  all  species
studied, the  progressive symptoms  typically found  after increasingly  higher
doses are irritation  of the mucous membranes, incoordination, mydriasis,  nar-
cosis1, tremors,  prostration, anesthesia,  and death.  Cats appeared  to be  more
resistant than dogs and rabbits; rats and mice are  less resistant than dogs  or
rabbits (Tables  12-1 and 12-2).
12.1.1.  Acute Exposure to Toluene
     12.1.1.1.   ACUTE INHALATION -- Carpenter et  al. (1976a)  reported 100?  mor-
tality in rats that were exposed for  4 hours  to  12,000  ppm of "toluene concen-
trate" (a mixture of  paraffins,  naphthenes,  and  aromatics that contained  45.9?
toluene and 0.06? benzene).   Treu.ors were seen in  5 minutes  and  prostration  in
15 minutes.  At  6300  ppm, inhalation  produced head tremors  in  1 hour and prostra-
tion in 2 hours, while only slight loss  of coordination was seen  after  4 hours'
exposure to  3300 ppm.  A calculated LC    of 8800  ppm  for a 4-hour period  of
inhalation of  "toluene  concentrate"  was reported.   Inhalation  of a  thinner
containing less toluene  (=33?) and only  0.01?  benzene  elicited  less  toxic
symptoms at a similar range of  doses in  rats in  a companion  study  by  the  same
laboratory (Carpenter et al.,  1976b).
     In a study  by  Smyth et  al. (1969a),  inhalar.ion of  4000 ppm technical  grade
toluene for 4 hours produced  1  death in  6 rats.   In an early  study.  Batchelor
(1927) noted  that inhalation of  1600  ppm of  toluene for  18  to 20 hours daily pro-
duced initial effects of instability and incoordination, conjunctivitis,  and
lacrimation,  and subsequently  narcosis  and mild  twitching.  A drop  in  body
temperature followed  by  death, occurred after 3 day.",  of exposure.  At  necropsy, a
severe cloudy swelling of the kidneys was  found.   In  this study, no effects  on
                                      12-1

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                                               TABLE  12-1

                                         Acute  Effects  of Toluene





ro
l
rv>
Route
inhalation
inhalation
inhalation
inhalation
inhalation
inhalation
4 nUnl -* *- , — —
Species
rats
rats
rats
rats
rats
rats
« n *• _
Dose
2^,^00 ppra for 1,5 h
12,200 ppm for 6.5 h
13,269 ppm
4,000 ppm for H h
12,000 ppm for 4 h
("toluene concentrate")3
8,800 ppm for 4 h
("toluene concentrate")3
£ *inr\ ~ 	 <*_ ~. h v.
Effect
60? mortality
50? mortality
Lethal dose
1/6 dead
Lethal dose
LC50
I I n « rJ *- _ «— , n _. n J -, 1 Vi
Reference
Cameron et al
Cameron et al
Faustov, 1958
Smyth et al . ,
Carpenter .et
Carpenter et

., 1938
,, 1938

1969a
al.,. 1976b
al., ?976b
-iT 1 nTC v.
                             ("toluene concentrate")''


inhalation     rats           3,300 ppm for l4 h
                             ("toluene concentrate")0

inhalation     rats           1,700 ppm for *4 h
                             ("toluene concentrate")0

inhalation     mice          24,^00 ppm for 1.5 h

inhalation     mice          12,200 ppm for 6.5 h

inhalation     Swiss mice     5,320 ppm for 7 h
                             (less than 0.01?
                              benzene present)
Prostration in 2 h, normal
  3 h after exposure

Slight loss of coordination    Carpenter et al.,  1976b
No-effect-level


10? mortality

100J mortality


LC50
Carpenter et al.,  1976b


Cameron et al., 1938

Cameron et al., 1938

Svirbely et al.,

-------
                                            TABLE  12-1   (cont.)
Route
inhalation
inhalation
inhalation
inhalation
Species
mice
mice
mice
rr t r •.
Dose
6,942 ppm for 6 h
(99. 5J purity)
6,634 ppm
9,288 ppm
8,600 ppm or 15,000 ppm
Effect
LC50
LSo
Lethal dose
SOJ reduction respiratory
Reference
Bonnet et al . ,
Faustov, 1958
Faustov, 1958
Carpenter et al

1979


., 1976b
 inhalation


 Inhalation
rvj
i
OJ
 inhalation

 inhalation

 inhalation



 inhalation
cats
("toluene concentrate")1

5,OUO ppra
("toluene concentrate")c

 7,800 ppra for 6 h
("toluene concentrate")'
guinea pigs   4,000 ppm for 4 h

rabbits       5,500 ppm

dogs            850 ppm for 1 h
dogs
760 ppm "toluene con-
centrate" t  h/d x 2 d
rested for 4 d, exposed
again for 3 d
  rate

No-effect-level for
respiratory rate

Progressive signs:  slight
  loss of coordination,
  roydriasla, slight hyper-
  sensitivity to light within
  20 min, prostration within
  80 min, anesthesia within
  2 hours.
  One death during 14 d
  observation period

2/3 dead within a few days

Lethal within 40 .min

Increased respiration rate,
  decreased respiration
  volume

Weight ]or,s of 1.1 kg in
1 of 2 dogs, otherwise normal
                                                                        Carpenter et al.,  1976b
Carpenter et al.,  1976b
                                                           Smyth and Smyth, 1928

                                                           Carpenter et al , ,  1944

                                                           Von Oettingen et al.-,
Carpenter et al . ,  1976b

-------
                                                TABLE  1?-1   (cent.)
 Route
               Species
inhalation -dogs
oral
            rats
                           Dose
                       1,500 pprc "toluene con-
                      centrate" 6 h/d x 3 d

                      7.53 g/kg (6.73 to 8.43)
                           Effect
                                                                                 Reference
                       51ight laci imatlon and head  Carpenter et  al.,  1976b
                       tremors (2 dogs exposed)
                                                             LDt
                                                    Smyth et  al . ,  19&9a
oral       Wistar rats
           adult

oral       Sprague-Dawley rats
           (150 to 200 g)

oral       rats
           14 d old, both sexes
           young adults
ro
£T





i.p.
i .p.
i .p.
older

rats
rats
rats
adults

and mice


i.p.       rats  (both sexes)
i.p.       mice (male)
                      7.0 g/kg
                      5.58 g/kg
                      (5.3 to 5.9 g/kg)
                      3.0 mil/kg (2.6 g/kg)
                      6.4 ml/kg (5.5 g/kg)
                      7.4 mi/kg (6.4 g/kg)

                      2.0 ma/kg (1.7 g/kg)

                      0.75 mil/kg (0.7 g/kg)

                      1.75 to 2.0 rat/kg
                      (1,5 g/kg to 1.7 g/kg)

                        800 oig/kg at 26°C
                        530 mg/kg at 8°C
                        255 mg/kg at 36°C

                      1.15 g/kg in olive oil
                      (1.04 to 1.31 g/kg)
                      (graded doses between
                      0.79 and 1.65 g/kg)
                       LD
                                                    50
                       LD50
                       LD
                       LD
                       LD,
  50
  50
                         50
                       Lethal dose

                       Apathy

                       Death from respiratory
                       failure

                       Approximate lethal dose
                       LD,
                                                                .0
                                                             Observed for 24 h
                                                             Cause of death:
                                                             respiratory failure
Wolf et al.,  1956


Withey and Hall, 1975



Kimura et al., 1971



Cameron et al . , 1938

Batchelor, 1927

Batchelor-, 1927


Keplinger et al.,  1959



Koga and Ohmiya,  1978
i.p.
mice (female)
g/kg
LD
                                                               50
Ikeda and Ohtsuji, 1971

-------
                                            TABLE 12-1  (cont.)
Route
i.p.
i .p.
Species
mice
guinea pigs
Dose
4 g/kg
2.0 mfc pure solvent
Effect
Lethal dose
6/10 dead after 2 h
Reference
Tsuzi, 1956
Wahlberg, 1976
s .c.
i.v.
dermal
(single
application)

dermal
dermal
derrcal
                 (1.7  g)

rats and mice  5  to  10 rod/kg
                 (4.3  to  8.2  g/kg)

rabbits        0.15  mi/kg (.13  g/kg)
               0.20  m!./kg (.17  g/kg)

rabbits        14.1  md/kg
rabbits        uncovered  application
               to  abdomen

rabbits        10  to  20 applications  of
                undiluted  toluene  to
                rabbit ear and  bandaged
                to shaved  abdomen
guinea pigs    1 IT.!,  for  16  h
                                                          All dead after 6 h
                                                          Lethal dose
                                                          13* mortality
                                                          100$ mortality
                                                          LD
                                                            50
                                                          Slight  irritation
                                                                         Cameron  et  al.,  1938
                                                                         Braier,  1973
                                                                         Smyth  et  al.,  1969a
                                                                                         Smyth et  al.,. -1969a
                                                          Perceptible  erythema,          Wolf et  al.,  1956
                                                            thin layer  of devitalized
                                                            tissue  which  exfoliated
                                                          No effect on  gross  appearance,
                                                            behavior, or  weight
                                                          Karyopyknosis,  karyulysis,
                                                            perinuclear edema,
                                                            spongiosis,
                                                            junctional separation,
                                                            cellular  infiltration  in
                                                            derrois,
                                                            no  liver  or kidney  damage
                                                                                         K.ronevi  et  al.,  1979

-------
                                                   TABLE  12-1  (cont.;
Route
dermal
   Specifis
guinea pigs
                                          Dose
                     Effect,
                                                                                         Reference
;\0 rot,  covered  Completely absorbed by 5th
                   to 7th d
                 No mortality up to 4 wk
                 Weight less than controls
                   for wk 1 to 3. no difference
                   at wk IJ
Wahlberg, 1976
to ,
i corneal
corneal
cCrneal
rabbits
rabbits
rabbits
C.005 -nl.
0.005 E!
: drc™
                                                      Moderately severe  injury

                                                      Moderately severe  injury

                                                      Perceptible  irritation  of
                                                         conjunctival membranes
                                                      No corneal  injury
                                                                      Smyth et al.,  1969a

                                                                      Carpenter and Smyth,  T9lifc

                                                                      Wolf et al.,  1956
h = hour; nin = minute; d r day; wV' : week; i.p- = irtrnperi toneal;  s.c. = subcutaneous;
i.v. = intravenous; n i number; ns = not specified
 "toluene concentrate" is 3 mlxt^re oT  paraffins, naj-lithcnea and arotr.aticp that contained ^5-9? toluene
 and 0.06% benzene.

-------
                                             TABLE  12-2

                                    Subchronic Effects of Toluene
Species  Route
            Dose
                        Effect
                                                               Reference
Rat      Inhalation  1600 ppm
                     18 to 20 h'd
                                    Initial effect of instabilitj-
                                    and incoordination,  conjunc-
                                    tivitis, lacrimation, and
                                    sniffles;  then narcosis
                                                               Batchelor,  1927
Rat      Inhalation  1600 ppm
                     18 to 20 h/d x 3 d
                                    Mild twitching;  drop in body
                                    temperature;  death.  Histology:
                                    severe cloudy swelling of
                                    kidneys, no effect on liver,
                                    heart, or testes
                                                               Batchelor,  1927
Rat      inhalation  1250 ppm
                     18 to 20 h/d
                                    Slight instability and
                                    incoordination;  mucous
                                    membrane irritation
                                                               Batchelor,  1927
Rat      Inhalation  620 ppm or 1100 ppm
                     18 to 20 h/d
                                    No-effect-level for symptoms;
                                    hyperpiasia of bone marrow
                                                               Batchelor,  1927
Rat
Inhalation
1000 ppm solvent mix-
ture (50 to 60$
oenzene, 30 to 35$
toluene, *l? xylene)
7 h/d x 5 d x ?b wk
No effect on body weight;
lymphopenia followed by leuco-
cytosis and lymphocytes is;  tran-
sient changes in blood picture
before or after each daily
expooure; splenic hemosiderosis
greater than that found after
inhalation of benzene only:
si ight to moderate reduction
2-1/2 wk after exposure.   Nar-
rowing of peri-follicular  collars
of cells in spleen,  no fat in
livers and kidney; iron-negative
pipjnent in kidneys of a few  animals.
Svirbely et al.,

-------
                                                 TABLE  12-2  (cent.)
   Species  Route
                 Dose
                          Effect
                                     Reference
   Rat      Inhalation       240, iJ80, or 980 ppm
                             "toluene concentrate"
                             6 h/d x 5 d/wk x 65 d
                                           No effect  on red  blood  cell
                                           count,  white blood  cell count,
                                           hematocrit,  hemoglobin, total
                                           and differential  white  count,
                                           blood urea nitrogen,  SuOT,
                                           SGPT, alkaline  phosphntase,
                                           or body weight.
                                                               Carpenter  et  al.,  1976b
   Rat
Inhalation
3184
4 h/d x 30 d
CD
Increased levels of SCOT,
SGPT, 0-lipop^oteins
Decreased levels of gluta-
thione, catalase, peroxi-
dase, total cholesterol
Khinkova, 1971
   Rat
Inhalation
200 ppm or 600 ppm
No narcosis; body weight
change in HBC count, RBC
count,or hemoglobin during
weekly sampling; increase in
percentage of segmented cells;
histological changes:   slight
pulmonary irritation;  few
casts in straight collecting
tubules in rats at 600 ppm;
no change in liver, spleen,
heart, or bone marrow
Von Oettingen et al,,

-------
                                                   TABLE  12-2  (cont.)
Species
Route
Rat
Inhalation
                 Dose
                              2500 ppra or 5000 ppm
                              7 h/d x 5 d x 5 wk
                              5 d/wk x 15 wk
                        Effect
                                  Reference
                        Transient decrease in body
                        weight;  hyperactivity,  marked
                        incoordination,  recovery after
                        cessation of exposure;  mor-
                        tality in 5000 ppm group 18/25;
                        increased bleeding time;  blood
                        •picture:   total  leucocytes
                        reduced after each exposure;
                        pulmonary lesions occurred
                        earlier  than in  group exposed
                        to 200 or 600 ppm;  casts in
                        renal tubules in all  rats
                        within 2  wk of exposure;  rest
                        of histology saine as  200
                        and 600  ppra
                        cytochrome P-J450,  ethoxy-
                        coumarin  0-deethylase increased;
                        UDP glucuronslytrarisf erase in-
                        creased only at  end of  exposure
                                  von Oettingen  et  al.,
Rats
Inhalation
CFY rats     Inhalation
(both sexes)
7 consecutive cycles
daily, 5 d/wk x 8 wk:
each cycle, 10 min to
12,000 pprc followed by
20 min toluene-free
internal
                 265 ppm 6 h/d x
                 5 d/wk z 1,  3 or
                 6 mo
Depression of body weight gain;
increased SCOT,  LDH levels;
no effect on BUN levels
Depression of kidney, brain,
and lung weights.  Histology:
no effect on brain, lung, liver
heart, or kidney, no sign of
lipid vacuolation in liver

Bromsulphthalein retention
decreased;  Cytochrome P-450
increased independent of
period of exposure; SGOT
and SGPT activity unaffected
Bruckner and Peterson,
198la
                                                         Ungvary et al.,  1980

-------
                                                  TABLE  12-2  (cont.)
    Specious
Rom e
            Dose
Effect
Reference
    CFY rats     Inhalation  929 ppm H h/d x
    1-bot n sexes)             5 d*'wi; x -i wk,
                             6 w-'  f. mo
ro
I
    CFY rats
    (.raaler)
Inhalation  398,  796,  1592  ppm
            8  h/d x  5  d/wk  x
            It  . . i .
                                    Cytochrome  P-150  increased
                                    independent  of  exposure
                                    period;  no  effect  on SCOT
                                    or SGPT;  aniline  hydroxylase
                                    and  aminopyrine N - dern ethyl as e
                                    activity increased; cytochrome
                                    bc concentrations  increased.
                                    KIstological  effects:
                                    dilation of  cisternae of
                                    rough  endoplasmic  reticulum;
                                    increase of  autophagous
                                    bodies which  was  dose and
                                    time dependent; retarded
                                    growth of females  but not
                                    males; glycogen content
                                    decreased

                                    Cytochrome  P-450  increased
                                    wi th dose
    Rat, guinea  Innalation  1G7 ppro continuously
    pig, clop,                for 90 d, cr 1085 ppm
    monkey                   8 h/cj, v d/wk x 6 wk
                                   Nc  effect  on  leukocytes, hemo-
                                   globin, or  hematocrit;  no  effect
                                   on  liver,  kidney, lungs, spleen
                                   or  heart;  ro  effect or.  brain or
                                   spinal  cord of  dogs and monkeys
                                                                          Jenkins  et  al .,  1970

-------
                                         TABLET12-2  (cont.)
Species   Route
Dose
Effect
                                      Reference
Mice      Inhalation  7 consecutive cycles
                     daily, 5 d/wk x 8 wk:
                     each cycle, 10 min. to
                     12,000 ppm followed
                     by 20 min. toluene-
                     free interval
                        Depression of  body weight  gain;
                        no effect  on LDH;  decreased  BUN
                        levels;  SCOT levels increased
                        (not  significantly) depression
                        of kidney, brain  and lung  weights;
                        Histology:  no  effect on  brain,
                        lung, liver, heart, or kidneys;
                        no sign  of lipid  vaculoation in
                        liver.
                                       Bruckner  and  Peterson,
                                       198la
Mice      Inhalation  4000 ppm 99.9% pure
                     toluene for 3 h
                        No effect  on  LDH  activity
                        significant increase  of
                        SGOT 24  h  post  exposure only
                                       Bruckner  and Peters-on,
                                       198lb
Mice     Inh?lation  4000 ppm 99.9$ pure
                     toluene for 3 h/d x
                     1, 3, or 5 d
                        SGOT  levels  increased after  1
                        and 3 days of  treatment;  no
                        effect 24 h  after  5 d
                                      Bruckner and Peterson,
                                       198lb
Mice      Inhalation  4000 ppm 99.9$ pure
                     toluene for 3 h/d x
                     5 d wk x 8 wk
                        Depression  of  body weight gain
                        during first 7 wk; increased
                        liver-to-body weight  ratio after
                        4 wk exposure, no effect at  1, 2,
                        or 8 wk;  no increase  in kidney,
                        brain,  or lung weights; SGOT activity
                        increased after  4 wk  of exposure
                        and  2 wk  post-exposure, but not
                        after 2 wk  or 8  wk of exposure;
                        no change in BUN.  Histology:  no
                        effect  on heart, lung, kidney,
                        brain and liver
                                      Bruckner and Peterson,
                                      198lb

-------
                                            TABLE  12-2 (cont.)
Species
Route
Dose
                                   Effect
                                                               Reference
Mice
Inhalation  1,  10, -100  or  1000  ppm  No  effect  on  body weight;  1
            6 h/d  x  20  d            and 10 ppm caused increase of RBC
                                   count on 10th day, recovery on
                                   day 20;  100 ppiu,  1000 ppm -
                                   decrease of RBC count; all doses -
                                   increase CJO  to 70?) of WBC count
                                   on  day 10,  recovery for all
                                   doses except  1000 ppm; 10 ppm to
                                   1000 ppm -  decrease of thrombo-
                                   cytes; histology:  100 ppm -
                                   Slight decrease in density
                                   of  bone marrow cells and in
                                   megakaryocytes and red cell
                                   elements;  1000 ppm - slight
                                   hypoplasia  of red cell elements,
                                   slight to  moderate disturbance
                                   in  maturity of neutrophils and
                                   thrombocytes, moderate increase
                                   of  reticulocytes, no change in
                                   brain, lung, liver, spleen,
                                   or  kidney.
                                                               Horiguchi and Inoue, 1977
Guinea pig  Inhalation  1250  ppm  4  h/d  x
                        6  d/wk  (18  exposures)
                        1000  ppm  4 h/d  x
                        6 d/wk  (35 exposures)
                                   Prostration, marked liver
                                   and renal degeneration,
                                   marked pulmonary inflammation

                                   No symptoms; slight toxic
                                   degeneration in liver and
                                   kidney
                                                               Smyth and Smyth,  1928

-------
                                                    TABLE 12-2 (cont.) .
    Species
Route
Dose
Effect
                                                           Reference
   Dogs
   2  experimental,
   1  control
Inhalation  2000 ppm 8 h/d x
            6 d/wk x l) mo, and
            then 2660 ppm 8 h/d,
            6 d/wk x 2 mo
                        Death on days  179 and  180; slight   Fabre  et  al.t  1955
                        nasal and ocular irritation; motor
                        incoordination and  paralysis of
                        extremities  during  terminal phase;
                        congestion in  lungs, hemorrhagic
                        liver,  reduced lymphoid follicles
                        and  hemosiderosis in spleen;
                        hyperemic renal glomeruli; albumin
                        in urine
   Dogs
ro
i
Inhalation  200,  400, 600 ppm
            3  8 h exposures
            for 1 wk, then 5 x 7 h
            for 1 wk and finally
            850 ppm for 1 hr
                        No effect  on  circulation, spinal
                        pressure;  increase of  respiratory
                        rate,  small increase of minute
                        volume,  decrease of respiratory
                        volume
                                   von Oettingen et al.f
   Dogs
Inhalation  400 ppm; 7 h/d x 5 d    Moderate temporary lyraphocytosis   von Oettingen et ai.,
   Rats
Oral        113 mg/kg/d,
            354 mg/kg/d,
            590 mg/kg/d x  T38
                        No effect  on
                        body and organ weights,
                        adrenals,  pancreas, femoral
                        bone marrow, lungs, heart,
                        liver,  kidney, spleen,
                        testes, bone marrow, BUN,
                        blood counts
                                   Wolf et  al.,. 1956

-------
                                                     TABLE 12-2 (cont.)
    Species    ~Route
                 Dose
                        Effect
                                  Reference
    Rats
Subcutaneous
1  cc/',:g x 21  d
Slight induration at injec-
tion site; 5 to 1H% loss of
body weight; transient slight
drop in RBC and WBC counts;
hyperplasia of bone marrow;
moderate hyperplasia of
malpighian corpuscle in spleen;
marked pigmentation of spleen;
focal necrosis in liver; slight
cloudy swelling in kidney;  no
effect on heart, testes, or lungs
Batchelor, 192?
    Guinea pig  Subcutaneous
                 0.25 cc/d x 30 to 70 d
I
Jr
                       Local  necrosis at injec-
                       tion site; survival period:
                       30  to  70 days; polypnea and
                       convulsions during last
                       days of survival; hemorrhagic,
                       hyperemic, and sometimes
                       degenerative changes in
                       lungs, kidneys, secondary
                       adrenals, liver, and spleen
                                  Sessa, 1948-

-------
                                                           TABLE 12-2 (cont.)
    Species
Route
Dose
                                                          Effect
Reference
Rabbit
i
~^
>ji
                     Subcutaneous      1 cc/kg x 6 d
                                        cc/kg
                                         Transient  slight -granulo-
                                         penia followed  by  ^ranulo-
                                         cyt6sis;  no change in bone
                                         marrow

                                         More marked effect on
                                         granulocytes; all  rabbits
                                         dead by end of  secord day;
                                         no effect  on bone  marrow
                                                               Braier,  1973
    h = hour; d = day; wk = week; SCOT = serun glutamic oxalacetic transarainase; SGPT = serum
    glutamic  pyruvic  transaminase; WBC = white blood cell; RBC = red blood cell; HDP = uridine 5'-phosphate;
    BUN =  blood urea  nitrogen; mo = month.

-------
the liver,  heart, or  testes were  observed,  although  hyperplasia of  the  bone
marrow was noted, suggesting possible contamination of the solvent with benzene,
     Cameron  et  al.   (1938),  found  that  24,400 ppm  of  toluene  produced  a
•nortality  of 60$  and  10?  in  rats and mice,  respectively, after  1.5  hours  of
exposure.   In  another group of  rats and  mice exposed  to one-half the  above
concentration  for a longer  period,  6.5  hours,  the mortality was  50$ and 100$,
respectively.   These two  species  are  probably equally sensitive.  Other studies
of mice include  that  of  S^irbely et al. (1943),  in  which the  7-hour   LC    in
Swiss mice was  determined to be  5320  ppz,  and that of Bonnet et al.  (1979),  in
which a 6-hour  LC  of 694? ppm  for mice   was noted.
     In the  study of  Carpenter et  al.  (1976b),  4 of 4  cats  initially  survi"ed
exposure to inhalation of  7800 ppm "toluene concentrate"  for  6 hours.   During
Qvprtcur-o »hp cats showed progressive si mi  of tcxicity, including slight loss of
coordination,  mydriasis,  slight  hyperdensitivity  to light  within  20  minutes,
prostration  within 80  minutes, and light  anesthesia within 2 hours.  Only 1 cat
died during  the 14-day observation period.
     /
     Inhalation of 4000 ppm toluer" (purified by distillation)  for 4 hours daily
was lethal within a few days to 2  of 3 guinea  pigs.. The third animal was  severely
prostrated.   Under the same regimen, animals  exposed to less than one-third of
this concentration (1250 ppm) for  6  days  a week  survived  3  weeks  of exposure,
although they were severely affected.  At 1000 ppm, guinea pigs  were not affected
even after 35 exposures,  although  there were slight  toxic degenerative changes
ia the liver and kidney (Smyth and Smyth,  1928).
     Carpenter  et al.  (1944)  reported that  inhalation  of a .concentration  of
about 55,000 ppm  was  lethal to  6  rabbits  in about  40 minutes  (range  of  24  to
62 minutes).
     Von Oettingen et al.  (1942b)  observed that  inhalation  of 650 ppm toluene
containing 0.01$  benzene for 1 hour by 6  dogs produced an  increase of respiratory
rate and a  decrease  of respiratory  volume.   Exposure  to  1500 ppn  of  "toluene
concentrate" for 6 hours daily for 3 days  produced only slight lacrimation and
head tremors in dogs.  Reduction of the concentration  of "toluene concentrate" to
1000 ppm did not alleviate tne  head tremors (Carpenter et al.,   1976b).
     Bruckner and Peterson (198la,b) found an age-dependent sensitivity in rats
anl Bice.  Mice, 4 weeks of age, were  found  to be more susceptible to exposure of
2600 ppm toluene vapor for 3 hours than 8 or 12 week old animals.
                                     12-16

-------
     12.1.1.2.   ACUTE ORAL  TOXICITY — An LD^  of 7.53 g/kg and 7.0 g/kg  body
weight for  a  single oral dose in rats has been reported by Smyth et al.  (1969a)
and Wolf et al.  (1956),  respectively.  Withey and Hall  (1975)  found 5.58  g/kg to
be the LDc0 in male Sprague-Dawley rats. Immature 14-day-old Sprague-Dawley  rats
were more sensitive than young  or  mature  adult male rats of  the same strain to
the acute effects of toluene (analytical grade)  in the studies of Kimura et al.
(1971).  These investigators determined an oral LD™ of 2.6 g/kg body weight, 5.5
g/kg body weight, and 6.4 g/kg  body  weight  for each group,  respectively.   This
age-dependent sensitivity  was   also  noted  by inhalation  exposure  (see  Section
12.1.1.1.).  Cameron et  al.   (1938),  however,  reported  that very young rats  were
more resistant to toluene than  adult animals of the Wistar strain.  Thirty-three
percent of  a  group of 12 nine day old rats  survived 5.25 hours  of exposure to air
saturated with toluene,  but 100$ mortality was observed in a group of adult  rats
exposed for the same duration.
     Based on the  results  of their  studies  on the oral toxicity of toluene in
animals of different age groups,  Kimura et  al.  (1971) suggested a maximum  per-
missible limit  of  0.002 g/kg bw for a single oral dose.   This was obtained by
dividing the  dose that  elicited the first  observable gross signs of CNS toxicity
by  1000.
      12.1.1.3.   ACUTE  EFFECTS  FROM INTRAPERITONEAL INJECTION  --  Mortality is
produced by a single intraperitoneal injection of toluene  in the range of 0.8 to
 1.7 g/kg  in  rats  and   mice.    Koga and  Ohmiya  (1978)  determined  an  LD^ of
 1.15  g/kg  body  weight  for  male mice from administration of  toluene graduated
 between 0.79  and 1.65 g/kg  and  diluted  in olive oil.  Respiratory failure was the
main  cause of  death in these  animals.   An LD™  of  1.64  g/kg was  reported for
 female  mice  by  Ikeda and Ohtsuji  (197D.   The reason for the disparity in the
 above   data  (e.g.,  interlaboratory  differences  or  sexual  differences in
 sensitivity)  has not been  ascertained.  In rats  an  i.p.  injection ->f 0.65  g/kg
 toluene  produced apathy, while 1.5 to  1.7 g/kg  produced  death from respiratory
 failure  (Batchelor,  1927);  1.7  g/kg was  a  lethal  dose in rats,  mice  (Cameron
 et  al.,  1938), and guinea pigs  (Wahlberg, 1976)*
      Savolainen  (1978)  observed that the concentration of radioactivity in the
                                                                            •)!(
 CNS was highest  in  the cerebrum  after an intraperitoneal injection  of methyl   C-
 toluene.  Label was iirrJetectable in the CNS by 24 hours post-injection,  which is
 consistent with the time course of clinical  signs of acute toluene intoxication.
                                      12-17

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    A  temperature-dependent sensitivity to  toluene  was observed in adult rats
of both sexes by Keplinger et al.  (1959).  The lethal  dose at 26°C was 800 mg/kg,
while  at  8°C  and 36°C,  lethal aoses were 530 mg/kg and 225 mg/kg, respectively.
The toxicity  of  toluene is greater in hot and cold environments.   It is unknown
whether increased susceptibility to toluene  is  caused by the stress of altered
environmental  temperature,   or  by  altered,  physiological   processes  (e.g.,
absorption,  diffusion,  distribution, or metabolic rate) .
     12.1.1.4.    ACUTE  EFFECTS  FROM   SUBCUTANEOUS   INJECTION  —  Subcutaneous
injection of 1.1 to 1.25  g/kg and 4.3  to 3.7 g/kg toluene have been reported to
produce mortality  in rats  and  mice,  respectively  (Batchelor,   1927;  Cameron
et al.,  1938).    Braier  (1973)  reported  that  4 cc toluene/kg  injected  into
rabbits produced marked transient granulopenia within 24 hours and marked granu-
locytosis ar.d ensuing death  in all animals by the end  of the second day.  A slight
area of induration was  seen  at the injection site.
     12.1.1.5.   ACUTE EFFECTS FROM  INTRAVENOUS  INJECTION —  Intravenous injec-
tion of 0.2  cc  toluene/kg produced 100? mortality in rabbits (Braier, 1973).
     12.1.1.6.    ACUTE  AND   SUBACUTE   EFFECTS OF  PERCUTANEOUS  APPLICATION  ~
Repeated application  of undiluted toluene to the raDbit ear or shaved skin of the
abdomen produced slight to moderate  irritation (Wolf  et  al.,  1956; Smyth et al.,
1969a)  ai;d  increased  local capillary  permeability  (Delaunay  et al.,  1950).
Continuous cutaneous  contact in  the guinea  pig resulted in slowed weight gain,
karyopyknosis,  karyolysis, spongiosis, and  cellular  infiltration in  the dermis
within  16 hours  (Kronevi et al.,  1979;  Wahlberg,  1976).   Application  to the
abdominal skin  of the   rat  produced   hemoglobinuria  (Schutz,  1960).    Slight
irritation of conjunct!val membranes but no corneal injury (Wolf et al.,  1956) or
moderately severe injury  (Carpenter  and Smyth,  1946;  Smyth et al., 1969a),  have
been observed following direct application of toluene to the eye.
12.1.2.   Subchrcnic  and  Chronic Exposure  to Toluene.   Subchronic  or chronic
studies of toluene have  not  indicated, with the exception of the high exposure
level study  of  Fabre  et al.   (1955), evidence of major toxic effects.
     Fabre ct al.  (1955)  exposed 2 dogs for 8 hours daily, 6 days  a  week, to
2000 ppm pure toluene via inhalation for 4 months, and subsequently to 2660 ppm
for 2 months.  Slight nasal  and ocular irritation occurred at the lower concen-
tration, and motor incoordination that  preceded paralysis of  the  extremities
occurred in  the  terminal  phase.  Death  occurred un days 179 and 180. There was no
effect on gain in body weight, on  the bone marrow, or  on  the adrenal., thyroid, or
                                     12-18

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pituitary glands.   Congestion in the lungs, hemorrhage in the liver, a decrease
of lymphoid follicles,  and  hemosiderosis in the spleen were  observed.  Glomeruli
of the kidney were hyperemic, and albumin was found in the  urine.
     Svirbely et al.,  (1944)  found that  repeated inhalation of  1000  ppm of a
solvent mixture containing  30  to 35% toluene,  50 to 60% benzene,  and a small
amount of xylene for 28 weeics  (7  hours/day.  5 days/week) had no effect on  body
weight in rats or  dogs.  There was no significant increase  of liver volume, and
•fat was not found  in the liver or kidneys; however, narrowing of perifollicular
collars  was  observed in the spleen  (Table  12-2).   Splenic  hemosiderosis was
greater than that  found ai'ter exposure to  benzene (Svirbely et al., 1944).
     Continuous exposure to 107 pptn toluene for 90 days or  repeated exposure  to
 1085 ppm toluene for 6  weeks (6  hours'day, 5  days/week)  did  not adversely  affect
the liver, kidney,  lungs, spleen, or heart in 30 rats,  30  guinea pigs, 4 dogs,  or
6 monkeys.  In addition, treatment-related effects were not  seen  in the brain  or
the  spinal  cord of  dogs or monkeys.   No  significant  change was observed  in
hematoiogic  parameters  (hemoglobin,  hematocrit, or  leucocyte  count).    All
animals with the exception  of  2  of 30 treated  rats  survived exposure, and all
 gained body weight with the exception of  the monkeys (Jenkins et al.,  1970).
     Repeated inhalation of 240,  460, or  980  ppm of "toluene concentrate" for
 13 weeks  (6  hours/day,  5 days/week)  produced no  treatment-related organ  damage
 in  rats  or  dogs.   Serum  alkaline phosphatase  (SAP),  serum  glutamic pyruvic
 transaminase  (SGPT), serum  glutamic  oxaloacetic transaminase (SCOT), and blood
 urea nitrogen (BUN) activities were normal.  Prior treatment  with  toluene did not
 render the  animals either   more  susceptible or more  resistant  to a subsequent
 challenge dose of  12,000 ppm (Carpenter et al., 19?6b).
     The  results of an unreviewec subchronic inhalation study with  rats that was
 performed  by Bio/dynamics  for  the   American  Petroleum  Institute  (1980)  are
 available.  Groups of 15 Sprague-Dawley  rats  of each sex were exposed 6  hours per
 day,  5 days  per week for 26 weeks to cumulative  concentrations  of 0,  100, and
 1481  ppm  toluene.   Initially,  the high  dose group was exposed to  2000 ppm, but
 the  dose  was  lowered to 1500 ppm after 7 exposures  because CNS depression was
 apparent.   A battery  of  blood and  clinical chemistry  tests  (BUN, SGPT,  SAP,
 glucose), urinalysis, and neurohistological examination of tissue was performed.
 The only treatment-related  sign was increased incidence of dry rales and staining
 of the ano-genital  fur  in the high level  treatment  group.   Significant changes  in
 the  blood and urine were not found with  the exception of  a dose-related decrease
                                      12-19

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in blood  glucose levels and a  dose-relatfcd increase  in SGPT levels  in female
                                                        »
rats.  Body weights were significantly higher in the high-dose male rats than in
the control rats, but this was not considered a toxic effect.  Treatment-related
neurohistopathological  changes  were  not  found.
     Inhalation exposure to 1000 ppcn  toluene for 6  hours  a  day, 5 days a week for
6 months  had  no  treatment-related  effects on male  OFA rats (Gradski  et al.,
1981).   Twenty-four rats/treatment  and  control group were  examined,  and body
weight  gain,  hematologic  parameters (BBC and -W3C  counts, hemoglobin,  mean
corpuscular  volume,  hematocrit,  sedimentation  rate),  and  tissue  histology
(lungs,  liver,  spleen,  kidney,  genitals,  and other  unspecified  "principal"
organs) were assessed.
     In  a chronic  inhalation  study conducted  by Industrial Bio-Test Labora-
tories, Inc. for the Chemical Industry Institute for Toxicology (CUT), groups of
120 Fischer  344 rats of  each  sex were  exposed to 30,  100,  or  300  ppm of high
purity  (>99.98J)  toluene  for  6 hours/day,  5 days/week  for 24 months  (CUT,
I960).   All  animals were weighed at  the beginning of the study, weekly for the
first 6 months,  every other week from 6 to 24  months,  and immediately prior to
sacrifice.   Hematology.  blood  chemistry, urinalysis, opthamology, and pathology
determinations were conducted  on randomly  selected  rats  that  were  sacrificed
after 6,  12, or 18 months to determine progression  of toxic effects (Table 12-3).
All remaining  rats were  sacrificed  for  study  after  24  months,  but  histopatno-
logical   examinations were  conducted only on  tissues  from  rats  in  the  high
exposure  (300  ppa)  and  control  (0 ppm) groups.
     Unscheduled deaths  occurred in 140 rats  (14.6J of  960 animals)  over the
2-year course  of the study, but mortality in the treated rats reportedly did not
differ significantly from controls (CUT, 1980).  The body weights of the treated
males were  found to be  significantly heavier  than the control  males throughout
 the  study,  although   a  clear   dose-response   relationship was  not  apparent
 (Table 12-4).    A  similar  effect was   noted  for   the  females  but  the  effect
 disappeared  during the  final  4  weeks of  the  study.   There were,  however,  no
 significant  differences  among  the  groups   in  absolute  organ  weights  (brain,
 liver,  heart,  kidneys,  lungs,  and testes or ovaries were weighed).  The battery
 Pf clinical chemistry  tests,  hematologic  studies  and  urinalyses  (Table   12-3)
 revealed  normal  levels  in the treated rats  except  for  two hematologic parameters
 in females.    Females exposed  to  100 or 300 ppm  toluene showed  slightly, but
 significantly, reduced  hematocrits,  and  the mean corpuscular hemoglobin concen-
                                     12-20

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                           TABLE  12^3

           Number of Rats Per Sex and Treatment Group
                    Examined at Each Interval
Interval
6 Months   12 Months  18 Months  24 Months
Hematologya
Blood Chemistry
Urinal ysisc
Ophthamology
r
Pathology
5
5
5
5
5
5
5
5
5
5
20
10
10
25e
20
10
10
10
58-66
68-76g
Hemoglobin   concentration   (HgB),   hematocrit   (HcT),   total
erythrocyte count  (RBC),  and  total  and  differential  leukocyte
counts (WBC) were determined; mean  corpuscular volume (MCV), mean
corpuscular hemoglobin  (MCH)  and mean  corpuscular  hemoglobin
concentration  (MCHC) were subsequently calculated.

 Blood urea nitrogen  (BUN), serum  glutamic pyruvic transaminase
(SGPT)  activity,  and  serum   alkaline  phosphatase  (SAP)  were
determined.
c
 Appearance, specific gravity,  protein or albumin, pH, ketones,
glucose, and presence of microscopic particles were determined.
d
 Ophthalmological  examinations  were   conducted  on all  animals
scheduled  for  interval  and final  (24-month)  sacrifice,  except
animals  also   scheduled  for  blood   collection  at  the  final
sacrifice.

 20 rats/sex/group  were  originally scheduled for sacrifice, but
an additional 5  rats/group  were examined  so  that  they could be
used  as  replacement animals should  any of  the  firit  group not
survive until  the final  sacrifice date.
f
 Histologic examinations were conducted on 38 tissues taken from
the high exposure  (300 ppm) and  control (0 ppm) rats  only.  All
scheduled  sacrifices, as well  as  rats that  were  sacrificed ir\
extremis (8-17 rats/sex/group) or  died during the  course of the
study (2-7 rats/sex/group),  were examined  grossly.
ff
 All surviving animals at the end of the 24 months were sacrificed
for pathologic examination.
                               12-21

-------
 tration was slightly, but significantly, increased in females exposed to 300 ppc
 (Table  12-5).    A variety  of  inflammatory,  degenerative,  prcliferative,  and
 neoplastic lesions were observed  in various tissues (see Section 14.1), but the
 lesions occurred  with equal  frequency in all control and treatment groups; CUT
 (I960)  concluded  that no tissue changes can be attributed to toluene inhalation.
 There were no  significant  differences among the groups; absolute ophthalsologic
 examinations did  not reveal any toluene-induced changes  in  the eyes of the rats.
        \
                                  TABLE  12-4
             24 Month  Chronic Exposure of Fischer 344 Rats Exposed
              6 Hours/Day, 5 Days/Week,  to Toluene  by  Inhalation

Group


Hales
Control '
30 ppm
100 ppm
300 ppm
Females
Control
30 ppm
100 ppm
300 ppm
i
•
Number
Animals


69
69
89
90

90
90
90
90

Mean
Body Weight in Grans
Weeks of
0


141
141
141
142

109
109
109
109
26


340
349«
3q 1.**
341

203
191"
194
195"
52


384
396"
140*4"
403"

213
211
211
211
Exposure
76


426
445"
4i47«»
446"

214
246"
248«»
248"
ICO


430
456"
4b4"
451"

260
272"
272"
271"
104


43C
454"
452"
445

265
273*
275
272
Total
Weight
Change

286
31U"
312«*
304"

156
164
166
163
 Source:  CUT,  1980
•Statistically significant  difference from control (P <0.05)
"Statistically significant difference from control (P <0.01)
            In  the   only  subchronic   oral   study.  female   rats   fed  up  to
590 mg toluene/kg by  intubation for  periods  of up to  6 months did  not  show ill
effects as determined by gross appearance, growth, periodic  blood counts, analysis
for  blood  urea  nitrogen,  final body  and organ weights,  bone  marrow  counts, or
histopathological examination  of adrenals, pancreas,  lungs, heart, liver, kidney,
spleen, and testis (Wolf  et al., 1956).
12.2.  EFFECTS ON LIVER,  KIDNEY, AND  LUNGS
            Toxic effects  in  the kidney,  and possibly in the liver and  lungs after
higher doses,  have been reported.
                                     12-22

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                                                        TAKLK  I. -•
                                                 Mematology Measurements
         21* Month Chronic Exposure of Fischer 344 Rats txpoaed  6  Hours/Day,  5  Days/Wvek,  to  Toluene  by  Inhalation
ro
Group

Control
30 ppm
100 ppm
300 ppm

Control
30 ppm
100 ppm
300 ppm

Control
30 ppm
100 ppm
300 ppm

Control
30 ppm
100 ppm
300 ppm
Number of
Animals

20
20
20
20

10
10
10
10

20
20
20
20

10
10
10
10
(103

6
9
6
6

7
8
8
7

it
4
3
4

4
5
5
4
WBC
/cu mm)

.03
.96*
.54
.53

.51
.66
.13
.50

.04
.59
.91
.21

.93
.40
.74
.87
,R3C
(10 /cu
18 Months
8.757
8.766
8.700
8.894
24 Months
9.666
8.736
9.925
9.407
18 Months
8.022
7.956
7.915
8.010
24 Months
8.397
8.274
8.076
8.090
HgB HcT
mm) (g/DL) (?)
of Exposure
16.56
16.61
16.47
16.80
of Exposure
18.91
16.58
18.47
18.33
of Exposure
15.67
15.77
15.75
15.78
of Exposure
16.46
15.89
15.94
15.86
(Males)
43.10
42.42
41.93
42.34
(Males)
51.78
46.51
51.61
47.35
(Females)
41.70
41.25
40.83
41 .20
(Females)
44.99
43.06
42.47»
42.02S«
MCV
(Cu. Mic.)

50
49
49
43

51
52
50
50

53
52
52
52

54
53
53
53

.4
.6
.5
.8"

.2
.5
.7
.9

.0
.8
.7
.4

.7
.3
-9
. 1
MCH
(we,)

18.87
13.90
IS. 91
18.35

19.24
19.05
18.67
19.44

19.49
- 19.77
I9.S5*
19.63

19.50
19. 11
19.68
19.52
MCHC

38.
38.
33.
39.

37.
36.
38.
39.

3T.
37.
38.
37.

36.
36.
37.
37.

04
82
93
30"

87
33
34
33

26
90«
24»
98

10
42
08
46»
      Source:  CUT,  1980
     •Statistically significant  difference  froa;  control  (P <0.05)
     ••Statistically  significant  difference  from control  (P <0.01)
     WBC = white blood  cell  count;  RGB  =  red blood  cell  count; HgB  ; henoglobin  concentration; DL =  100 milliliters;
     HcT = hematocrit;  MCV = mean corpuscular  volume; Mic. = micron; MCH = me.nn  corpuscular- hemoglobin; MCHC -
     mean corpuscular hemoglobin  concentration.

-------
12.2.1.  Liver
    Histological damage was not observed after subchronic or chronic inhalation
of 1000  ppm  of a solvent  mixture containing  30 to  35?  toluene  for  26 weeks,
980  pfffl  of "toluene concentrate"  for  12  weeks,  1085  pptL of toluene for f weeks,
or 300 ppm of 99-98? pure  toluene for 24 months in a variety  of species in the
studies  described in Section 12.1.2. (Svirbely  et al.,  1944;  Carpenter et al.,
1976b; Jenkins  et  al.,  1970; CUT,   I960).   Furthermore,  no  liver damage was
detected in female rats after daily oral doses of 590 mg/kg toluene for 6 months
(Wolf et al.,  1956).
    Two preliminary  reports  (abstracts) from  the  laboratory  of  Bruckner and
Peterson noted no effect on hepatorenal function.  In a regimen designed to zinic
solvent  "sniffing", male rats and mice were  exposed  to 12,000  ppm toluene for 7
ten-minute   periods  (interspersed   with   20-minute  toluene-free   periods),
5 days/week  for  8 weeks.   No organ pathology was  found.   Lactic dehydrogenase,
SGPT activity, BUN  level,  and liver  triglyceride  content  were normal  (Bruckner
and Peterson,   1978).    In  another  study,  inhalation  of  4000 ppm  toluene
(3 hours/day, 5  times weekly) for up  to 8 weeks failed to reveal toluene induced
hepatorenal  injury as  indicated  by  a   battery  of  toxicological tests  (SCOT
activity, BUN level, urinary glucose  and protein concentration, and urinary cell
count),  and  histopathological  examination  of  the  liver,   kidney,  and  lung
(Bruckner and Peterson, 1976).
    Intraperitoneal  injection  of reagent grade toluene  (corn  oil  vehicle) at
doses of 150, 300,  600, or 1200 mg/kg had no effect  on serum ornithine carbamyl
transferase  activity  in adult  male  guinea  pigs, when assayed  24  hours  after
injection  (Divincenzo and  Krasavage, 1974).   Histological  examination revealed
no liver abnormalities  or  lipid accumulation with the  exception of the highest
dose, where there was evidence of .lipid  accumulation.
    Two hours  after   male  rats  (weighing  150  to 300  g)  were  administered
239-5 mg/100 g body weight of toluene  in mineral oil  by  gavage,  there was no
.evidence of injury  to the  microsomal  function of  the liver.  There was no effect
on protein  synthesis,  cell sap RNA,  glucose 6-phosphatase, oxidative demethy-
lase, nicotinamide adenine dinucleotide  phosphate  (NADPH), neotetrazolium reduc-
tase, or lipid conjugated diene content of microsomes (Reynolds,  1972).  Inhala-
tion of 3CO ppm  toluene  (6 hours/day,  5 days/week)  for   15 weeks  slightly
increased cytochrome P-450 content in tlie liver, appreciably enhanced ethoxycou
oarin o-deethylase, and at the end  of exposure,  increased  UDP glueuronyltrans-
                                     12-24

-------
ferase activity.   The com.pnr,  of  .toluene in  osrirenal fat  tended to  decrease
during continued evDo.snre.  wMle presence in  the!  brain was detected  throughout
exposure.  The d.lr"ini'at-ion  r,f  toluene content in perlrenal fat at  the same  tine
that  drug met.?ho.i .< r.i.nc  pn^vrnps increased suggests an  adaptation to continued
presence  of the soi.vpr.t-.  fFV>«sara  et  al..  1979).
                ntpnpono  contact  with a dose  of  1.7  g (2.0 i-.i) toluene, whicn
was  completely  ahso'-he'l  "'vti-'in  5  to- 7 days,  .produced  no  change  in  liver
i"orpho5 ogy  (Wahlberc,  lpr-e ?^e others  that suggest  a  slight toxic  effect.  In  a
sU^y  by von Oettjpcpri f-f ;•! . M^Pb), inhalation of 600 to 5000 ppm toluene that
contained 0.01S ben-7pne  for S wpeks  (7 hours/day,  5  days/week)  by rats caused an
enlargement  of the  ] ivpr  f increase of  weight and  volume) in a  dose-dependent
manner To hours after the last exposure.  Histologically there  was a progressive
decrease- of  cytoplasm density  in the liver  cells  as  the concentration of toluene
        i
increased, but hyperemia was not noted.   These observations were not seen in rats
sacrificed  2 wee^s after the ]ast  exposure.   Matsumoto et al . (1971) reported an
increase in  liver wei eht and  liver weight  to body weight ratio  in  rats exposed
9 hours/day. 6 days/week for ^3 weeks  to 2000 ppm toluene  vapor.   This was not
noted  at lower doses (100  pr-m  or 200 ppm).
     In the  inhalation  study of  Fabre et al .  (1955),  2 dogs exposed for U months
(8 hours/day,  6 days/week)  to  2000 ppm  pure  toluene  and,  subsequently,  to
?£66 ppm for 2 months .  hari hemorrhagic livers.
     Tahti-   et al.   (1977)  observed  that   inhalation  of  1000  ppm  toluene,
8 hours/day  for 1  week, increased SCOT  and SGPT activity  and  induced metabolic
acidosis in  rats.
     Although  early  reports from the same laboratory  revealed  no effect on SCOT
activity or  BUM level in mice  or rats,  a recent report  by  Bruckner and Peterson
(198lb) noted  an  increase  in SOOT  activity in these species  during intermittent
exposure to  12,000  ppra  toluene  (Section 12.3.).  Increase in LDH  activity  was
seen  in rats  and decrease  in  BUN  levels  was  seen  in  mice.   .No  histological
changes were observed,  but an  increase of organ weight  to  body weight ratio was
found.
     Histological  changes  in   the  liver  were  found  when  male  CFY  rats  were
injected  intraperitoneally with O.U3  or  0.87 g/kg  body  weight of  analytical
grade   toluene   for  up to  ^ weeks  (Ungvary et al . ,  1976).  There was  a dose-
                                     12-25

-------
dependent increase in the number of mitochondria per unit of cytoplasmic area in
the liver.  Total area,  nuclear density,  and nucleus/cytoplasm ratio increased at
the higher  dosage.   Dose-dependent decreases  in nuclear volume were seen after
intraperitoneal  or  subcutaneous  injection,  witn  subcutaneous  injection being
less effective than  intraperitoneal injection.  The  authors suggested that the
considerable  accumulation of mitochondria  was  related  to increased  metabolism by
the liver,  and that  oxidative  detoxification of the solvent might  involve mito-
chondrial  enzymes as well  as  hepatic microsomal  enzymes.  In an earlier paper,
Ungvary et  al. (1975) found that  intraperitoneal or aubcutaneous administration
of  toluene  produced degenerative  changes, i.e.,  separation of  ribosomes  and
vacuolar dilation of the rough endoplasmic reticulum.   In  these  studies,  the
higher  concentrations  of  toluene-also  decreased  glycogen  content.   Following
discontinuation of exposure,  the  hepatic  changes indicating  increased  load on
detoxification  processes   (increased  succinate  dehydrogenase  (SDH)  activity,
increase of mitochondria and  smooth  endoplasmic  reticulum,  decreased glycogen
content)  as   well   as  degeneration   (dilation  of  endoplasmic  reticulum,
accumulation of autophagous  vacuoles)   rapidly  regressed,  indicating  that  the
toxic  and  liver loading effects  of  toluene are  reversible.   The regenerative
property of  the  liver after  hepatectomy  was  not  significantly affected by
exposure to toluene  (Hudak  et  al., 1976).
     In a more recent study by Ungvary et  ai.  (19&U),  male CFY rats were exposed
via inhalation to 265 ppm  (6  hours daily),  929  ppm  or  1592 ppm (8 hours daily)
analytical grade toluene, and  female  rats  were exposed to the lowest dose only (5
exposures a week  for up to 6 months).   Growth was  inhibited in  males  at  the
higher concentrations,  and  in  the females.  No abnormal nistological changes  were
found  in the liver, but liver weight  was increased  by treatment.    Signs of
adaptive compensation  included proliferation of  smooth endoplasmic reticulum,
increased  cytochrcme   P^50  and   cytochrorae  b,  activity,  increased  aniline
hydroxylase activity,  and  increased  aminopyrine N-demethylase activity.  These
 changes were  dose-dependent and reversible, but showed little or no  dependence on
length of exposure.  There  was no effect on SCOT or SGPT activity.  The authors
 concluded from their latest studies  that  subchronic  exposure to toluene vapors
has  no specific hepatotoxic  effect.    The  histological effects  of inhalation
 exposure to toluene  were corroborated by the earlier intraperitoneal or subcuta-
neous  studies,  with the  exception  that  necrotic areas were not found after
 inhalation.  Whether or  not this reflects  the different  route of exposure or the
                                     12-26

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higher concentration  of  toluene  administered  intraperitoneally has  not been
ascertained.
12.2.2.    Kidney.   Histological  effects  of  renal toxicity  were not  seen  in
subchronic inhalation studies (Table 12-2) mice exposed to  1000 ppm for 20 days
(Horiguchi and Inoue,  1977)i  in  rats,  guinea pigs, dogs, or monkeys exposed to
1085 ppm for 6  weeks  (Jenkins  et al., 1970), in rats and mice exposed to 4000 ppm
toluene for  8  weeks  (Bruckner and  Peterson,  1981b),  or  in chronic inhalation
studies in rats exposed to 300 ppm for 24 months (CUT, 1980).  Toluene did not
elicit an observable effect in renal histology after daily subchronic oral  dosing
at a level of 590 mg/kg for 138 days in rats  (Wolf  et  al.,  1956).
     Pathological renal changes,  however,   have been observed in somfe studies.
von Oetvir.^en et al. (1942b)  found increasing numbers of casts  in  the collecting
tubules of rat kidneys during inhalation of concentrations  ranging from 600 ppm
to 5000 ppm for 5 weeks (7 hours  daily,  5  days/week).   A few casts in the  kidney
were seen after the third week of exposure at 600  ppm  and earlier in the  higher
doses.   Appreciable fat  in   the  convoluted  tubules  and  hyaline casts  in the
collecting tubules were observed in dogs following inhalation  exposure  to  200 to
    i
600 ppm for approximately 20  daily 8-hour  exposures, subsequently to 400 ppm for
7 hours/day,  5 days/week  for  1  week,  and  finally  to  850 ppm for  1 hour.
Matsumoto et al. (197D reported  that inhalation exposure to toluene at a level
of  2000 ppm for 8 hours/day,   6 days/week for  43 weeks  produced hyaline droplets
in  the renal tubules  of  rats.  There was an  increase  in  kidney  weight and the
ratio of kidney weight to  body weight.
     Inhalation  of  2000  ppm toluene  8 hours/day,  6 days/week  for  4 months,
followed  by exposure  to  2600 ppm  during  the remaining  2  months,  produced
hyperemic  renal glomeruli and  albuminuria in  dogs  (Fabre et al.,  1955).   In
guinea  pigs,  inhalation  of  1000  ppm  distilled   pure  toluene  (4 hours/day,
6 days/week for  a  total  of 35 exposures)  produced  slight toxic degeneration :n
the kidney  (Smyth  and  Smyth,  1928).  Eighteen  exposures  at a higher levels of
 1250 ppm produced more marked degeneration.   Degeneration of  convoluted tubular
epithelium  in  guinea  pigs  exposed to  toluene by the subcutaneous  route was
reported in an  abstract of a  paper  by Sessa  (1948).
 12.2.3.   Lungs.   Histological  lung  damage  was not  seen  after  inhalation of
 1000 ppm  toluene  for  20 days   in  mice  (Horiguchi  and  Inoue,   1977),  after
inhalation  of   1085 ppm  for   6 weeks  in rats,  guinea  pigs,  dogs.,  or monkeys
 (Jenkins et al., 1970), after inhalation of 4000 ppm for 8 weeks in rats and mice
                                      12-27

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(Bruckner  and Peterson,  198lb), after inhalation of 300 ppm for 24 months in rats
(CUT,  I960), or  after daily ingestion of 590 mg/kg for  133  days  in rats (Wolf
et al.,  1956).
     Irritative effects on the respiratory tract,  however,   have  been reported
(Browning, 1965;  Gerarde,  1959;  Fabre  et al.,  1955;   von  Oettingen  et al.,
19t2b).  Marked pulmonary inflammation was seen in guinea pigs after inhalation
of  1250 ppm  distilled  pure  toluene  for  ^ hours  daily,   6 days/week,  for
18 exposures  (Smyth  and Smyth, 1928).   Hemorrhagic,  hyperemic,  and sometimes
degenerative  pulmonary changes  were observed in  guinea pigs after a subcutaneous
injection of  0.22 g of toluene  daily for 30 to 70 days  as  reported in an abstract
(Sessa.  1948).    Repeated  exposure  to  concentrations of  200  to  600 ppm toluene
produced congestion  in the lungs of dogs,  and  pulmonary lesions were elicited in
rats  after  1  week  of  inhalation   of   2500 ppm   (7 hours/day,   5 days/week)
(von Oettingen  et al., 1942b).  Congestion in  the lungs was noted by Fabre et al.
(1955) in dogs  exposed for 8 hours a  day,  6 days a week to 2000 ppm toluene for
4 months,  and subsequently to 2660 ppm for 2 months.
12.3.  BEHAVIORAL TOXICITY AND CENTRAL NERVOUS SYSTEM EFFECTS
     Excessive  depression of the CNS  has been associated with acute exposure to
high levels of  toluene.  A concentration of 20,000 ppm  toluene was lethal to rats
after 30  to 50 minutes  of  exposure, with  death attributed to depression cf the
CNS  (Kojima and Kobayashi, 1975,  cited in NRC,  1980).   Inhalation of 12,000 ppm
of "toluene concentrate"  containing 0.06?  benzene  was lethal to rats following
tremors  that   appeared  within  5  minutes  of  exposure,   and prostration  that
occurred within  15 minutes of exposure (Carpenter et al., 1976b).
     A  dose-related  effect  on instability,  incoordination, and  narcosis  was
found in rats exposed 18 to 20  hours daily to  toluene  concentrations of 1600 ppm
and  1250 ppm (Batchelor, 1927).;  no symptoms were seen at 1100 ppm.   Carpenter
et al.  (1976b)  reported that  rats  were  unaffected  by  inhalation  exposure  to
1700 ppm  of  "toluene  concentrate"   for  4 hours,  and   suffered  only  slight
incoordination  at 3300  ppm.   Dogs were  unaffected by exposure  to 760 ppm for
6 hours,  but  exhibited head  tremors at  1500 ppm.   After  inhalation of 7800 ppm
"toluene  concentrate" for 6 hours, cats exhibited loss of coordination followed
by prostration  and,  finally^ light anesthesia within 2 hours, but no mortality.
     Bruckner and Peterson  (198lb) observed  that  the  onset of narcosis and the
depth of  CNS depression was dose-dependent in mice exposed via  inhalation  to
                                     12-28

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12,000 ppm,  5200 ppm, or 2600 ppm toluene.  Recovery was rapid.  Aftf>: exposure
to 12,000 ppm for 20 minutes, mean  performance levels scored prior to exposure
were restored within approximately one-half hour in 1-week-old rats.
     A single intraveriqus injection of 0.06 g  toluene per kg body weight caused
generalized rigidity with  hyperextension of the  back within  10  seconds  in an
experiment  with  1  dog  (Baker  and Tichy,  1953).   Recovery  occurred  within
12 minutes.  When a series of  10 doses  of 0.06 g toluene/kg  was given intra-
venously  every  3  t°  5  days  to another  dog, rigidity  and  twitching  of the
extremities were induced.  Recovery occurred in 5  to 10 minutes.  At necropsy,
cortical  and  cerebeliar atrophy was  found.   Marked shrinkage  and  hyperchro-
maticity  of many cortical  neurons,  patchy myelin  pallor  and   fragmentation,
especially  in  perivascular   areas,  were  found.    Multiple  fresh  petechiae,
especially in the white matter, were seen.  There was  a decrease and  degeneration
of Purkinje cells in the cerebellum (Baker and Tichy, 1953).
12.3.1.   Effect of  Solvent-Sniffing Abuse.  In the section on  CNS effects on
humans (Section 11.1.), inhalation of  readily available  thinners  by  young adults
has  been described  as  a prevalent practice  that  typically  affects  the  CNS.
Inhalation  of  solvent  mixtures  containing toluene in  the  laboratory rat  have
       i                                                  ,            1
demonstrated similar effects.  Inhalation of a mixture of solvents  containing 25?
methylene chloride, 5% methanol, 1)3% heptane, and 23% toluene for  10 minutes (60
to 226 mg/Jl) caused a  decrease  in grooming,  the appearance of ataxia, abnormal
scratching, hind  limb  flaccid  paralysis and,  finally,  unconsciousness in male
Fischer  rats.  Cumulative effects were noted  with 4 intermittent  10-minute  expo-
sure  periods with 15 minutes  between exposures.  When the interval  between each
exposure was increased  to  40 minutes,   recovery  was  almost  complete  (Pryor
et al.,  1978).
      Subchronic exposure to a thinner  containing toluene impaired acquisition of
a complex behavior.  Rats  inhaled 50,000 ppm  of a readily available  commercial
paint thinner composed of 42?  toluene, 25? methanol, 10? methyl  iso-butyl ketone,
and  minor amounts of other solvents for 4,  8, or  16 weeks  (twice-daily for 10
minute periods, 5 days  a week) and then were observed for acquisition of temporal
 discrimination in  a  differential reinforcement  of low rate schedule (DRL 20).  In
 this  test,  the animal  is  rewarded for a  bar press  separated  from the last
 response by 20  seconds.   These results suggested that  persistent inhalation of
 thinner  vapors  impaired  temporal  discrimination when  the  animals were tested
 within a relatively  short  time after  the period   of inhalation.   However,
                                      12-29

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responses in rats that had a period of  rest  after exposure did not differ from
controls (ColoUa et al., 1979).
     Studies in  laboratory  animals have shown that  toluene  contributes to the
symptoms of thinner toxicity.   In  the studies  of Peterson and Bruckner (1978),
impairment of cognitive function and muscular coordination were used to monitor
CNS  depression  and narcosis]    Behavioral   performance  (visual  placing,  grip
strength, wire  maneuver,  tail  pinch, and  righting  reflex)  in mice exposed to
3980 pptn toluene for 3 hours decreased over  time of  exposure, and was inversely
correlated with toluene concentration in brain  tissue.  Concentration of toluene
in the brain increased exponentially with  the  length of exposure and similarly
decreased after  termination of  exposure, as  did  levels of toluene in liver and
blood  (Figure  12-1).   A  single 10-minute exposure  to a  higher concentration
(10,615 ppm) was consistent with  the  pattern elicited  by  the  lower concent-
ration, longer duration  exposure.   Recovery  of behavioral performance occurred
as solvent  concentration in the brain  decreased  after exposure.  Bruckner and
Peterson  (198la)  noted that ataxia, immobility in  the absence of stimulation,
hypnosis with difficult  arousal and unconsciousness  were apparent in mice with
    i
blood  concentrations  of HO  to  75 ^ig/g, 75  to  125  ng/g,  125 to  150 ng/g and
>150 ng/g,  respectively, as measured by the air  bleb method.
     A study was conducted by Peterson and Bruckner (1978)  with mice to mimic the
conditions  typical  of  human solvent-sniffing abuse.   Intermittent exposures to
10,615 ppm  for  approximately  3  hours (5  minutes  of   exposure  followed  by
10 minutes  without,  toluene, or 10 minutes  of exposure  followed by 20 minutes
without  toluene),  or  to 11,9^2 ppm  for approximately  3  hours  (10  minutes  of
exposure  followed  by  20  or 30  minutes  without toluene)  were conducted. Reflex
performance became  progressively  lower  throughout the  experimental  periods  in
the  regimens that included toluene-free intervals of 20 minutes or less,  A 30-
minute,  toluene-free  interval  between  exposures  permitted  almost  unimpaired
perfrrmance,  indicating  complete  recovery  between  exposures   (Peterson  and
Bruckner, 1978).
     In a later acute study, Bruckner and Peterson (198la)  exposed mice and rats
to  seven consecutive  cycles.   Each  cycle  consisted  of  10-minute  inhalation
exposure  to 12,000 ppm toluene followed by  a  20-minute,  solvent-free recovery
period.  Unconditioned performance and reflexes of the  animals were tested prior
to and following exposure.   The  mice showed almost complete recovery during the
                                     12-30

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                                     TISSUE LEVELS
                   tu
800-,

600-


400-

ioo-
                                                             —I
                                                              4
    0     1     2    3     t     2    3
    HOURS OF EXPOSURE  HOURS POSTEXPOSURE
                      100 -,
                     o
                     er
                     ui
                     0.
                                 NORMALIZED TISSUE LEVELS
                                               	— BRAIN
                                               	LIVER

                                                  	BLOOD
                                    I
                                    1
    0    1
    HOURS OF EXPOSURE
            II
            3
                                              HOURS POSTEXPOSURE
                   UJ
                   z
                   UJ

                   O
                    o
600-


400-


200-
                                 BRAIN CONCENTRATION VERSUS
                                CHANGE IN PERFORMANCE SCORE

                                             	 BRAIN
                                              	APERFORMANCE
                               I     I     T
                          0123

                          HOURS OF EXPOSURE
                               i
                              2
                    -5  u,
                     4  2

                        re
                      _ IL.
                     1  a
                    r 1  uj
                     b~ °"
            3    4
HOURS POSTEXPOSURE
Figure 12-1.    Toluene  Levels in Tissue and Behavioral Performance (Mice were con-
              tinuously  exposed for  3  hours to an  intoxicating  concentration cf
              3980  pprn toluene.  Groups  of animals were  analyzed  for air bleb con-
              centration,  reflex  performance,  and  tissue levels after 15,  30, 60,
              120,  and 180 minutes  of exposure  and 1,  2,  and  t(  hours postexposure.
              Figure  12-1A shows toluene levels in liver, brain, and  blood.  Figure
              12-1B shows  toluene  normalized  to the  highest  mean  level  in each
              tissue.  Figure 12-1C compares brain levels of toluene  with change in
              performance  of the animals.  Lines represent means.  N = 7 mice on all
              but  H hours  postexposure,   in  which   case,  N  =  6.   (Pt-.terson and
              Bruckner,  1978)
                                      12-31

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course of  treatment,  while performance  scores  of rats exhibited  a  progressive
decline.   The  authors  speculate that ,the rapidity of  recovery  in mice might be
attribu'.ed to  the higher circulatory, metabolic,  and  respiratory  rates of mice;
the increasing CNS depression seen  in rats  over a 3-hour  period of intermittent
inhalation might result  from  a  progressive accumulation of the chemical.   Sub-
stantial residual quantities  in the  brain 1 hour post exposure had been noted by
the same authors in ah earlier  paper (Bruckner  and Peterson,  196la).
     In a subchronic study, groups of 6  mice or  U  rats with comparable numbers of
controls were  subjected  to 7  consecutive cycles  (as described  in  the  preceding
paragraph) on  a daily  basis,  5  times a  week for 8 weeks (Bruckner and  Peterson,
 198la).  Depression of body weight gain was  observed in both rats and mice during
the  8  weeks  of intermittent  toluene exposure.   An increase  in SCOT levels was
noted  in  rats and  mice,  but  the  increase  in mice  was  net   statistically
significant.   An increase  in LDH was seen in rats at all sampling  intervals, but
 this  effect  was not  noted in  mice.   BUN levels in rats were   unaffected  by
 treatment, whereas BUN levels in mice were  consistently lower during the period
. of exposure.  Recovery occurred  2 weeks  after exposure.  There were no effects on
 brain, lung, liver, heart, or kidney histology, although a depression in gain of
 organ  weights  (kidney, brain, lung) was noted in  both species.
 12.3-2.   Effects  on Simple and Complex  Behavioral Performance.   After a single
 exposure  to 800 ppsn  toluene for   4 hours,  unconditioned  reflexes  and  simple
 behavior  (corneal,  grip,  and righting  reflexes,  locomotor activity,  and coor-
 dination)  began to  fail  (Krivanek and Mullin,  1978; Mullin and  Krivanek, 1982).
 In these studies, male rats were exposed to concentrations of  0, 800, 1600, 3200,
 and  6^400  ppm and tested  at 0.5, 1,   2, and  k hours during  exposure and 18 hours
 after  exposure (Table  12-6).
     Concentrations   of   toluene  as low  as  1  ppa   administered  6  hours/day
 depressed  wheel  turning performance (a spontaneous  activity) after 10 days of
 exposure  in adult  male  mice  (Horiguchi  and Inoue,   1977).   No effect  on body
 weight was seen at  any of  the dosages used  (1,  10, 100, and 1000  ppm)  during 20
 daily  exposures.  However, alterations-in blood elements were  observed in animals
 exposed to 10, 100, or 1000 ppm (Table  12-7).
     The positive findings at 1 ppm  reported by Horiguchi  and  Inoue (1977), and a
 report of changes in motor nerve chronaxies  in rats exposed continuously to ^ ppm
 toluene  for 85 days  (Gusev,  1967;   cited by NRC,  1980).  are at  variance with
 negative effects observed  in  other  experiments at much higher levels and may be
                                      12-32

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                                                                              TABLE  ^-6


                                                                     Behavioral  Effects of Toluene
 I
U)
txl
Species
Wistar rats

Sprague-Dawl ey
rats

Rats (scale)

Rats



Rats (oalej


Rats (male)


Rats (eile)


Sprague-Dawiey
rata
Mice


Mice

Mice (sale)

Mice
Mice (male)





Route
Inhalation

Inhalation


Inhalation ,

Inhalation



Inhalation


Inhalation


Inhalation


Inhalation

Inhalation


Inhalation

Inhalation

Inhalation
I. p.





Dose
571*, 1118, 2296, and
1595 ppa
150 ppm for 0.5, 1 , 2 or
« h

550 to 800 ppu for
« h/d x 2 wk
"4000 ppto 2 h/d x 60 d



3UOO ppm for ->4 It
1000 ppm for b h

3200 ppm for "4 h

1600 ppa for 1 h
800 ppm for 1 h


23,000 ppm for 1/2 h/d x 7.6 d

3980 ppn for 3 ^
10,615 ppa for 10 mln

4,000 ppo for 3 h/d x
5 d/wk for 8 wk
1, 10, 100, 1,000 ppa for
6 h/d- » 10 d
2650 ppm
0.96 g/kg





Effect
Deficit In multiple
response schedule
Initial stimulation
followed by depression In
multiple response schedule
No effect on avoidance
response
Multiple response schedule
No effect on CRF or FR30
Deficit In DHL 12 sec
schedule
Deficit In conditioned
avoidance response
No effect level
Deficit conditioned
avoidance response
No- effect- level
Deficit in unconditioned
reflexes and simple
behavior
Induced forced turning

1/eficlt in visual placing,
grip strength, wire maneuver
tail pinch, righting reflex
Deficit on an accelerating,
rotating bar
Deficit in wheel-turning

Causes mice to fall on side
Loss of righting reflex in
5/7 In 20.6 + 1.6 Bin
Interval froo loss of
righting reflex to re-
covery 35.0 + 8.2 nin.
1t.3J lethality in 2M h
Reference
Colotla et el.,
1979
Celler et al., 1979


Bat tig and Grandjean, 1964

Ikeda and Mlyaka, 1978



Shlgeta et al., 1978


Krlvanek and Hullin, 1978;
Mullin and Krivanek, 1982

Krivanek and Mullin, -1978
Hullin and Krivanek, 1982

Ishlkawa and Schaldt, t973

Peterson and Bruckner,
1978

Bruckner and Peterson,
1976
Horiguchl and Inoue, 1977

Fausto», 1956
Koga and Ohalya, 1978





        r s hour; d = day; wk 5 week; i.p. =  Intraperltoneal; Bin = minute; sec = second.

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                                                TABLE 12-7

                                     Myelotoxicity Effects of Toluene
      Species
Route
Dose
Effect
Reference
      Rats
     Rats,
     Guinea  pigs,
     Dogs,
     Monkeys
     Rats
I
(A)
Inhalation  200,  600,  2500 or
            f.OOO  ppm for 7 h/d,
            '5  d/wk x 5 to 6 wk
Inhalation  107  ppm  continuous
            exposure for  90  d,
            or  1085  ppm  for  8 h/d,
            5 d/wk  x 6 wk
Inhalation  30,  100  or 300 ppra  for
            6 h/d x  5 d/wk x 24 mo
     Rats,
     Dogs
Inhalation
240, 480, or 980 ppm
for 6 h/d x 5 d/wk
x 65 d
A temporary decrease of
lympho-ytes and total at
the highest doses
white blood cell count;
no anemia; no effect on
bone marrow or spleen
No significant change in
leukocyte count, hemo-
globin, or hematocrit

No effect on any hemato-
logical parameter except
2 parameters in females:
at 100 or 300 ppm hemato-
crit was reduced, at
300 ppm mean corpuscular
hemoglobin concentration
was higher; no histo-
pathology on any organ
including spleen and bone
marrow
No effect on red blood
cell count, white blood
cell count, hematocrit,
hemoglobin, total and
differential white count,
SAP, SGPT, SCOT, or BUN;
no effect on bone marrow.
                                                    Von Oettingen et al., 1942b
                                                    Jenkins et al.,  1970
                                                    CII'i,  1980
Carpenter et al., 1976b

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                                           TABLE 12-7 (cent.)
     Species   Route
            Dose
                        Effect
                               Reference
      Rats
Inhalation
200, 1000 or 2000 ppm
for 8 h/day x 32 wks
ro
t
oo
Ul
      Rat
      Rats
Inhalation
Oral
112 ppm for
4 h/d x ij mo
Significantly retarded weight
gain at 2 higher doses during
initial 4 wks; no significant
difference in hemoglobin
hematocrit and total plasma
protein; no significant in-
crease of RBC; significant
increase of leucocytes at
highest dose at first week
of exposure followed by
recovery; eosinophile counts
decreased rapidly in the
first 2 to J) weeks and then
recovered; increase of
Mommsen's toxic granules.
Leukocytosis and chromo-
some damage in bone marrow
118, 354 or 590 mg/kg/d Normal bone marrow,
x 138 d                 spleen, bone marrow
                        counts, blood count
Takeuchi, 1969
Dobrokhctov and
  Enikeev, 1975
(cited in U.S. EPA, 1980b)

Wolf et ai.,  1956

-------
                                            TABLE 12-7 (cont.)
     Species   Route
              Dose
                        Effect
                                                   Reference
     Rats
     Rat
     Rat
    tlice
u>
     Dogs

     Dogs
Subcutaneous  0.8?  g/kg/g
              x  1U  d

Subcutaneous  1  g/kg/d  x 12  d
Dermal
Inhalation
Inhalation

Inhalation
10 g/kg/d

1, 10, 100 or 1000 ppm
for 6 h/d x 20 d
                    Normal leukocyte count,
                    spleen, and bone marrow

                    11.5$ chromosome damaged
                    cells vs.  3-9? in controls
                    Impaired leukopoiesis
                    Leukocytosis at all dose
                    levels;  100, 1000 ppm:
                    depressed red cell count;
                    10 to 1000 ppm:  decreased
                    thrombocyte count;
                    1000 ppm:  trend toward
                    hypoplasia in bone marrow
ppm for 7 h/d x 5 d No change in blood picture;
                    temporary lymphocytosis
2000 ppm for 8 h/d x
6 d/wk x i\ mo, and
then 2600 ppm for 8 h/d
x 6 d/wk x 2 mo
                    No effect on bone marrow
Gerarde, 1960

Lyakalo, 1973

Yushkevich and Malypheva,
   1975
Horiguchi and Inoue, 1977
                                                       Von Oettingen et al.,
Fabre et al., 1955
     n = number; h = hour; d = day; wk = week;  mo = month;  SAP =  serum alkaline phosphatase;
     SGPT = serum glutamic pyruvic transaminase;  SCOT -  serum glutamic oxalacetic transaminase;  BUN = blood urea
     nitrogen.

-------
regarded as spurious. For example, Takeuchi et al. (1991) reported that exposure
to 1000 ppm toluene  for  16 weeks  (12 hours/day) did  not  produce  evidence of
peripheral  (tail) nerve injury (as determined by nerve condition velocity, mixed
nerve  conduction velocity,  and  distal latency  measurements)  in Wistar rats.
Ikeda  and Miyake  (1978)  did  not find any  effect on  spontaneous  activity in
studies  of  repeated  exposure  to   4000 ppm  toluene  in  rats.   However,  the
behavioral  tests of  the  latter  authors  were  carried out  4 days  after final
exposure,  and  rapid  recovery of behavior after  exposure  (Shigeta et al., 1978;
Peterson and Bruckner, 1978; and Ishikawa and Schmidt, 1973) may explain in part
the disparate  results.
    A  single exposure  to  3000 ppm  toluene  for U hours  disrupted  established
timing  of  bar  pressing in a  conditioned avoidance response test with adult male
Wistar  rats (Shigeta et al., 1978).  Concentrations of 0 and  1000  ppm toluene did
not affect  this operant behavior. At 3000 ppm, increased response and shortening
of the inter-response-interval  were noted,  but  no  change  in  shock counts was
seen.   Behavioral  recovery  occurred  1 hour  after exposure.  Krivanek and Mullin
(1978)  reported  a  decrease  in  conditioned  avoidance reflexes in male rats after
inhalation of  3200 ppm toluene for  & hours,  but they reported no effect at dose
levels  of 1600 or  800  ppm.
     In another  study of operant  behavior, Colotla et al., (1979) used rats  that
had been trained to reinforced  bar pressing in a multiple schedule that consisted
of fixed ratio  (FR)  10  and differential  reinforcement of  low rates  (DRL) 20-
second  components with a 60-second time out between reinforcement periods.  Five
trained adult  Wistar rats were exposed to  concentrations of 57^, 1H»8, 2296, or
^595 ppm toluene,  and  test sessions  were  36 minutes  long.   Control  sessions
intervened between solvent  exposure  sessions to assess recovery.  A decrease in
rate of response of the FR component  and an increase of  freouency  rate of  the DRL
component were observed with all doses in  a  dose-dependent manner.  No residual
effects were observed.
     Exposure  to a lower  concentration of  toluene (150 ppm) for periods of  0.5,
1, 2,  or H hours affected performance  on a  multiple  fixed ratio-fixed interval
schedule of reinforcement in  ;  male Holtzman Sprague-Dawley rats.   An  initial
enhancement of FR  and FI  rates occurred during the shorter  exposure periods,
followed  by a  decrease in rates  during longer  exposure periods  (Geller  et  al.,
1979);  however, only a small number of animals was used, and  the response was not
uniform.   Battig and Grandjean (1961!) found  no effect on acquisition or  consoli-
                                     12-37

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dation of an avoidance response in 6 adult male rats after inhalation of toluene
that varied  in concentration from 550  to  800  ppm,  fpr 4 hours/day for 2 weeks.
Continued exposure at similar levels for another week  effected a somewhat slower
extinction of  the avoidance response.
     Repeated  exposure  of rats  to inhalation of 4000 ppm toluene, 2 hours daily
for 60 days, did not  affect spontaneous  locomotor activity,  emotionality, or
learning on  2  operant schedules:   memory in a continuous reinforcement schedule
(CRF) where every bar press was  rewarded by food, and extinction  of a fixed ratio
                     i
(FR 30). schedule performance  where  every  30th  response  is reinforced.   The
exposure impaired  learning on  a  third  operant  schedule,  however,   in  which
acquisition  of a differential  reinforcement of a low rate of responding (DRL 12
seconds) schedule required rats to allow at least 12 seconds  between responses to
receive a reward.  Impaired performance was present 60  days after final exposure.
Exposure to  toluene  appears  to affect more seriously higher  levels of cognition.
Histoiogicai examination  of the  brain did not  reveal any changes  (Ikeda and
Miyake,  1976).
     Inhalation of  1*000 ppm toluene by mice for 3 hours/day, 5 times weekly for
up to 8 weeks  caused a steady deterioration of performance on an accelerating,
rotating bar curing  the initial hour of each session of exposure. Solvent levels
in blood and liver  increased during each exposure session and decreased quickly
after exposure (Bruckner  and Peterson,  1976).
     Circling  (forced turning)  was produced  within  a mean of 7.6 days in 90-day-
old  male  Sprague-Dawley  rats  (n=10)   by  repeated  exposure  r.o  approximately
23|000 ppm  (4  to 5 mi. in kO to 50 4. of  air) for one-half hour per day.  After 15,
21, or 3*1  days of recovery, the rats were  reexposed daily  to toluene.  When only
 15 days of  recovery had  elapsed,  the  number  of  exposures  required  t'o elicit
forced turning was  significantly  let>s  than the nuinter required to acquire the
behavior originally. This effect was  not seen when a  longer period of recovery
had elapsed.  Thus,  toluene has a residual effect and  the effect is reversible.
This turning  was  not  associated  with  any  histological  lesions  in  the  brain
 (Ishikawa and  Schmidt,  1973).
 ^•So.   Effect on  Electrical  Activity of  the Brain  and  Sleep.  The effect of
 toluene on electrical, as well  as  behavioral, parameters in the brain was studied
 by Contreras et  al.  (1979).  Twenty cats were exposed via trachea.l cannulation to
7000 to  52,000 ppm (approximate) concentrations of  toluene for 10-minute periods
 a day, 7 days  a week for *40 days; exposures  were  started at 7000 ppm and were
                                     12-38

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increased  by  increments  of approximately  7000 ppm  (with  10-minute recoveiy
intervals  between  exposures) each  10 minutes until  electrical  and  behavioral
changes  appeared.   During the first seconds of acute intoxication at  12,000 ppm,
the behavior consisted of restlessness, polypnea, coughing;  sneezing, and vege-
tative responses consisting  of salivation, mydriasis, and lacrimation.  Itaxia
appeared 2 minutes later, ending with postural collapse.  Changes of electrical
actisvity at this point were  found  in the anterior lobe  of  the cerebellum, the
amygdala,  and  the  visual cortex.   There was no  behavioral  responsa to light,
sound, or pain stimuli  (Table  12-8).  The threshold  dose for restlessness war,
approximately 7,000 ppm.  No behavioral response to external  stimuli  occurred  at
approximately  39,000 ppm.   Recovery  from  ataxia   occurred  12 minutes  after
removal from  exposure.   With  repeated exposure at  a concentration of approx-
imately 27,150 ppm,  hypersynchronous rhythms spread  from the amygdala  to the
reticular formation,  visual cortex, and  cerebellum, and  electrical activity
appeared in the gyrus  cingvli  which coincided with  a display of hallucinatory
behavior.  These EEC and behavioral signs are similar to complex partial seizures
in man (Contreras  et al., 1979).
     Takeuchi and  Hisanaga   (1977,) found  that 1030,   2000,  or UOCO  ppm toluene
administered for 4  hours  to groups of 4 or 5 male Wistar rats  elicited changes  in
the  sleep  cycle,  altered cortical  and hippoceapal ERG  rhythms,  and increased
pulse rates.  All  phases  of  sleep  were disturbed at  concentrations  of 2000 and
4000 ppm;  1000  ppro deterred the onset  of the  slow-wave phase of  sleep,  but
facilitated onset  of the paradoxical phase.
     A similar observation was made  by Fodor et al.   (1973), where an increased
percentage of REM during  sleep was found in female albino rats during  exposure  to
 1000 ppm.  A  concentration of  1000 ppm decreased cortical and hippocampal com-
ponents of the EEC (Takeuchi and Hisanaga, 1977).  Exposure  to 2000  ppm toluene
increased cortical fast  components and hippocampal components, whereas exposure
to  1)000 ppm  increased the hippocampal  fast   component  as well.   At 4000 ppm,
excitability  measured  by rearing  reactions   (standing  on hind legs) increased
during the first hour of  exposure, but  this  phase was followed by a depression
and  the  rats  were unable to stand  or walk;  excitability increased again upon
reexposure.   At  2000 ppm only  increased  excitability  was observed,  and   at
 1000 ppm excitability was not increased significantly.  Myoclonic seizures were
seen in both  2000  and 4000  ppm  treated  groups,  with  greater frequency at the
higher concentration.
                                      12-39

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                                                       TABLE 12-8

                                          Central Nervous System Effects  of Toluene
   Species
 Route
                Dose
                                                             Effect
  Cats
                 Inhalation  ca. 7,000 to 52,000  ppm
                               10 min/d x 40 d
  Rats
Inhalation  1000, 2000,  or ^000 ppm
              for H h
ro
Jr
O
  Rats (male)    Inhalation  2000 ppm  toluene  for
                               8 h/d x 1  wk
  Sprague-Dawley Inhalation  500 ppm 6 h/d x 3 d
  Rats (male)                  Killed 16 to 18 h  after
                               exposure

                             1000 ppm 6 h/d x 5 d
                               decapitated 4 h after
                               exposures
                                                                                           Reference
  Rats
Inhalation  1000 ppm x 6 h/d x
            6 d/wk x J! wks
  Rats,  mice     Inhalation  265  ppm
Restlessness
Autonomic nervous system
  stimulation, ataxia,
  collapse
EEG changes
Seizures

EEG changes
Increased excitability
Changed sleep cycle
Increased pulse rate

Decreased threshold for
  Bern egride-induced
  convulsions

Increase of catecholamines
  in lateral palisade
  zone of median eminence

Increase of catecholamines
  in subependymal layer of
  median eminence
Increase of FSH

Increased spontaneous activity
during light period after
repeated exposure.   Single
exposure did not influence
cir^adian rhythm.

Threshold affecting CNS
                                                                       Contreras et al.,  1979
Takeuchi and Hisanaga,
  '.977
                                                                       Takeuchi and Suzuki,
                                                                          1975
                                                                       Andersson et al.,  1980
Ikeda et al.,  1981
                                                                       Faustov, 1958
  min =  minute;  d =  day;  h =  hour; -wk =  week;  EEG - electroencephalogram; FSH - follicle-stimulating hormone;  CNS =  central
  nervous system.

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     Convulsion  threshold after  intraperitoneal  injection  of Bemegride  was,
decreased significantly by preexposure to 2000 ppm toluene  for 8  hours/day in
6 Sprague-Dawley tale rats.  The change was noted after 1 week  of exposure, and
convulsion threshold continued to decrease  over 6 weeks of  exposure.   After 8
weeks of exposure,  the difference from the controls was not significant, although
the convulsion threshold remained lower.  The data suggest that toluene renders
the CNS more  susceptible  to  induction of a convulsion state.  Body weights of
these rats were lower than those of controls  during the exposure  period, although
differences were not significant (Takeuchi and Suzuki,  1975).
12«3.^«  Effect on Neuromodulators.  Andersson et al.  (1980) reported an increase
of dopaioine and noradrenaline in the median  eminence after inhalation of 500 ppm
and 1000 ppm toluene, respectively,   by male  rats.   The higher levels also pro-
duced  increases  of noradrenaline turnover  within the  median  eminence  and the
anterior periventricular and  paraventricular hypothalamic nuclei.  A significant
increase of plasma levels  of  follicle-stimulating hormone  (FSH)  and a non-signi-
ficant elevation of prolactin and corticosterone were  also noted.
   i  Yamawaki  et  al. (1982)  found  a  decrease  in specific  serotonin ( H)-5HT
binding to synaptic membrane fractions from whole brains, and from the hippo-
campus and pons/medulla oblongata regions of rats that were exposed  15 minutes a
day to 7000 ppm toluene for  14  days.   These results indicate that serotonergic
mechanisms may have  contributed to  some  of  the observed behavioral effects of
exposure  (i.e., hindlimb abduction,  resting tremor, head weaving).
 12.3.5.  Minimal Effect Levels.  Although most acute  as well as chronic studies
indicate  minor effects  of toluene  at  concentrations under  1000 ppm  and most
reviews  (NIOSH,  1973; U.S. EPA,  1980a;  NRC, 1980; Benignus,   198la, 198lb) have
emphasized the negligible effects of toluene  on the  CNS at this level, several
foreign  studies   suggest  that  lower  level  exposures may  not be  innocuous.
Horiguchi and  Inoue  (1977) found  a  decrement in simple task performance during
exposure  to   1 ppm  toluene,  Gusev  (1967)  reported  lengthened  motor  nerve
chronaxies at 4 ppm,  Colotla  et al.  (1979) noted a  decrement  in  operant behavior
at concentrations of 57*J ppm, and Anderson et al.  (1980) reported histochemical
changes in the brain at 500  ppm.  In all of these studies, sensitive parameters
of  CNS activity were measured.   Higher concentrations tended to  affect more
complex tasks. Furthermore,  the studies of Andersson et al. (1980) indicate that
500 ppm affects an area of the brain that regulates many vegetative, as well as
reproductive,  functions.   Although the  results of  the lower exposure level
                                      12-41

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studies  (Horiguchi  and  Inoue,  1977;  Gusev,  1967)  are   inconclusive,  these
findings indicate that  effects of toluene' on  the  CNS at levels below  1000 ppm
cannot be totally ignored,
12.4.  EFFECTS ON OTHER ORGANS
12.4.1.  Blood-Forming Organs.  Myelotoxicity is an  effect  that has been attri-
buted to toluene.  Prior to the early 1940's it was  believed  that toluene had the
same  effect  as benzene;  however,  in most  of the earlier  studies  toluene was
contaminated with benzene.  Since then, there have been studies indicating a lack
of myeiotoxicity, although several  have indicated a positive effect (see Table
12-7).
     One of  the first  studies  that  used pure toluene was that of von Oettingen
et al.  (1942b). Exposure  01'  rats  to 200 to  5000 ppm toluene contaminated with
less  than  0.01J  benzene  for 5 to  6 weeks  (7 hours/day,  5 days/week)  did not
affect blood-forming organs,  as indicated by the absence of anemia and changes in
the  bone marrow and spleen.   Exposure to the higher  concentrations of 2500 and
5000 ppm did produce a  daily temporary shift in the  blood picture  that  was
characterized by  a decrease  of lymphocytes and total white blood count, with a
moderate increase of segmented cells (Table  12-9).   Exposure of dogs to 400 ppm
toluene on 5 consecutive days for 7 hours daily  produced no appreciable changes
in the blood picture,  with the exception of a temporary lymphocytosis at the end
of exposure  (von Oettingen et al.,  1942b).   Fabre  et  al. (1955) also found that
exposure of  dogs  to  high concentrations of  toluene  containing less  than 0.1J
benzene  (2000  ppm   for  8 hours  daily,  6 days   weekly   for   4 months,  and
subsequently 2600 ppm  for the 2  remaining months)  had  no  effect on  the bone
marrow.
     Wolf et al. (1956) could find no  effect  on femoral bone marrow, spleen, bone
marrow  counts,  or hematological  parameters  in female Wistar rats orally dosed
with 94.4J pure toluene at levels of up to  590 mg/kg/day for 24 weeks.  Exposure
of  Fischer  344  rats for  24  months  (6  hours/day,  5  days/week)  to 30,  100,  or
300  ppm 99.98? pure toluene did not have any hematological  effects (CUT, I960)
 (see Table 12-4); there were also no changes in  the  bone marrow or spleen.
     Male Wistar  rats  administered  a  daily subcutaneous dose of 0.87 g/kg body
weight  for   14 days had  a normal leucocyte  count,  thymus  and  spleen weight,
femoral  marrow  nucleated cell count,  and  femoral marrow  nucleic acid content
 (Gerarde, 1956).
                                      12-42

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                          TABLE 12-9

Weekly Blood Picture of formal  Rata  and Rats Exposed to 600 and
   2500 ppn of Toluene 7 Hours/Day,  5 Days/Week, for 5 Weeks
'

Weeks
Preexposure period:
First

Second
Exposure period:
First

Second

Third

Fourth

Fifth

2 Weeks After
Exposure

Preexposure period:
First
Second
Exposure period:
First

Second

Third

Fourth

Fifth

2 Weeks After
Exposure

Preexposure period:
First

Second
Exposure period:
First

Second

Third

Fourth

Fifth

2 Weeks After
Exposure


Nuaibei of Animals

5
15
20

—
20
20
20
20
20
20
20
20
20
9
9


15
5
20

—
20
—
20
__
20
20
20
20
20
10
10


10
10
20

20
20
20
20
20
20
20
20
20
20
10
10



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 \
     Speck and Moeschlin  (1968) noted  that  subcutaneous  injection  of  300  or
700 rag/kg pure toluene administered daily to  rabbits for 6 and 9 weeks, respec-
tively,  had no myelotoxic effects.   There were no  changes  in DNA-synthesis  of
bone marrow cells as measured by incorporation of ^H-methylthymidine or1 in peri-
pheral  blood  elements  (leucocytes,  thrombocytes,  reticulocytes,  or  erythro-
cytss).
     Braier (1973) reported  that  subcutaneous  injection of  862 mg/kg toluene
daily for 6 days produced a moderate depression of granulocytes during the first
2 days of treatment.   This was  followed  by a sharp  rise in granulocytes by the
.eno  of  6 days  (i.e.,  twice that  of  the pretreatment  level).   No  significant
change was  noted  in  the bone  marrow.   In contrast, subcutaneous  injection  of
benzene  at the same dosage  elicited a progressive decrease in granulocyte count
throughout the period  of treatment.   Andrews et al. (1977)  found  that benzene
                               59
inhibited the incorporation of   Fe into erythrocytes  of  mice,  although intra-
peritoneal injection  of toluene alone did not (see Section 15.1).
     The studies suggesting a  myelotoxic effect include that  of Horiguchi  and
Inoue (1977), who exposed groups  of  6 male mice  to  toluene vapor at  concentra-
tions of 1, 10, 100,  or 1000 ppm for 6 hours  daily over a period of 20.days and
found that  the 2  highest  doses decreased red  cell count.   Concentrations  of
10 ppm and above decreased  thrombocyte count.  All  groups showed an increase  in
white cell count midway in  the  study,  followed by recovery except in the 100 ppm
group.  Slight hypoplasia of the bone marrow  was noted  at the highest dose.
     Takeuchi  (19&9)  observed  a  transient increase in leucocytes in 6 Donryu
strain rats exposed to 2000 ppm 99-9? pure toluene  containing less  than 0.2 ppm
benzene  in- the course  of  8-nour  daily  exposures  for  32  weeks,  as  well  as  a
transient decrease of  eosinophile  counts  upon exposure to 200, 1000, or 2000 ppm
toluene  under  the  same regimen  (see  Table  12-7).   After 32 weeks  of toluene
exposure, all groups  including an  unexposed  control  group  were subjected to  39
eight-hour  daily exposures to  benzene prior  to  sacrifice and histopathological
examination.  The-adrenal weight to body weight ratio was depressed significantly
in  all   groups  that  had been  exposed to  toluene.    Histologically,  the  zona
glonierulosa of the  adrenal cortex  of toluene-exposed rats was thicker, while the
zona fasciculata and  zona reticularis were reduced.   The authors suggested that
toluene  affected  the  hypothalamo-pituitary-adrenal   system.    While  that
hypothesis  is  tenable, since  the rats  exposed to  toluene differed  from  the
unexposed controls, all groups exposed and unexposed  to toluene were also exposed

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abstract of a later  paper (Takeuchi et al., 1972), which  was not available for
review, noted that exposure of male rats to toluene for 8 hours daily for ^ weeks
increased adrenal weight and eosinophil counts and decreased corticosteroid con-
centration after  1 week.
     Topical  application  of 10 g/kg toluene to  rats  4 hours daily for 4 months
had no effect on maturation of erythroblasts in the bone marrow, but an increase
of plasmic and lymphoid reticular cells in the marrow  indicated an impairment of
leucopoiesis.  A  lower aosage  of 1  g/kg toluene daily had no effect (Yushkevich
and Malypheva,  1975).
     Leukocytosis and  chromosomal  damage in the bone marrow (Section 1^.2.3-3.)
was noted  in rats that had been exposed  via inhalation to  112 ppm  of toluene,
4 hours  daily for  4 months  (Dobrokhotov and  Enikeev,  1975).    Recovery  from
leucocytosis  occurred  1 month after termination  of exposure, but the chromosomal
damage was unchanged.  It was  also  reported that inhalation of a combination of
toluene  and  benzene  produced chromosomal aberrations,  which were approximately
equal to the sum  of aberrations induced by single administration of the solvents.
Further; benzene  caused leukocytopenia, but the  mixture caused leukocytosis.  It
should be  noted,  however, that  the results  of  this study should be regarded as
inconclusive  because Russian  reports  of toluene-induced chromosomal aberrations
have not been corroborated by western investigators (Section 1i(.2.3.?.).
     In the  studies  by Katsumotc et al.  (1971), exposure of Donryu male rats to
inhalation  of  2000 ppm  toluene vapor  8 hours/day,  6 days/week for  43 weeks
decreased  the ratios of  thymus weight to body weight  and  spleen weight to body
weight.
     Although the evidence tends to weigh more  heavily toward the absence of a
myelotoxic effect from toluene exposure in animals,  the  suggestion  made by NEC
 (1980) that  the positive  findings may  indicate subtle  unrecognized hematopoietic
responses  is  sound.  For  example,  the effect of tcluene  on  hematocrit and mean
corpuscular  hemoglobin concentration  in  female  Fischer rats  and not in male rats
 (CUT, I960)  is of interest in view of the observation of Hirokawa (1955), where
there appears to  be  a  higher  susceptibility of  the  female  rabbit  to benzene.  In
that study,  the decrease  in erythrocyues,  hemoglobin content, white blood cells
and mean corpuscular hemoglobin concentration,  and increase in mean corpuscular
volume in the female was simulated in the  estradiol  propionate-treated orchi-
dectomized male.
                                     12-45

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    There  was  no  increase  of erythrocyte  fragility  in 6 rats  that inhaled
20,000  ppn  "toluene concentrate" for  45 minutes (Carpenter et al.,  1976b).   A
slight  increase in coagulation  time was noted  in  rabbit blood by Fabre et al.
(1955)  and  in  rats  by von Oettingen et al. O942b).
12.4.2.  Cardiovascular Effects.  Several  animal studies  have shown that massive
doses of  toluene cause a number of electrocardiographic changes*  In addition, a
sensitization  of the  heart to low oxygen levels has been observed.
     Inhalation of glue  fumes  containing toluene for  1  minute  significantly
slowed  sinoatrial  heart rate and slightly lengthened  the P-R interval in 8 ICR
mice.   .Subjecting  the animals to 5 minutes of  asphyxia  after inhalation of the
glue fumes  produced a 2:1 atrioventricular block in all animals  within  an average
of  42  seconds  of  asphyxia.    In  contrast,  after  24  five-minute periods  of"
asphyxia, the  sinoatrial  heart  rate rose,  the P-R internal  did not lengthen, and
atrioventicular (AV)  block did  not  occur in 12 mice (Taylor and Harris, 1970).
     In acute inhalation of  toluene,  atrial  fibrillation, bradiarrhythmia, and
asystole, along with respiratory paralysis, occurred.  Subcutaneous injection of
two doses  of  0.87  g/kg  body weight  daily for  6 weeks  elicited repolarization
disorders,  atrial  fibrillation,  and  in some  of  the  rats,  ventricular extra-
systoles  (Morvai et al.,  1976).
     Intravenous injection of 0.01  mg/kg epinephrine into dogs following inhala-
tion of  toluene vapors  elicited  ventricular  fibrillation  (Chenoweth,  1946).
This observation is of interest, because  the  "sudden  death" ayndrome following
"glue sniffing" in  humans might possibly be explained by an increased secretion
of epinephrine, which could  cause  fibrillation of  the heart as a result of the
combined  effect of  the two compounds.
     Intravenous injection of 0.5 mg/kg body weight of toluene into rats reduced
arterial  blood pressure;  however,  injection of the same dosage  by  the intra-
peritoneal or  subcutaneous  route  had no effect  on blood pressure (Morvai et al.,
1976).   No effect  on  blood pressure was seen in the chronic inhalation study of
von Oettingen et al.   (1942b),  where dogs were  exposed  to inhalation of 200 to
600 ppm toluene several  times  weekly  for several  months.   In this  study,  no
effect  was  observed  on  blood  pressure,  heart rate,  venous  pressure,  spinal
pressure, respiratory rate, minute  volume, or respiratory volume.
12.4.3.  Gonadal Effects.   Matsumoto et al. (197D  found  that exposure of Donyru
strain  male rats to  inhalation of  100 or 200 ppm toluene  vapor 8  hours/day,
6 days/week for 1  year produced no  change in  erythrocyte and leucocyte counts,
                                     12-46

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 serum total  protein,  or cholinesterase activity.   However,  at  the higher dose,
 degeneration of germinal cells of the testes was found in H of 12 animals while
 normal  germinal epithelium was found in controls.   Testicular  weight  was lower
 than controls  at  both  dose  levels.   There was a trend toward  a  decrease  of
 testicular to body weight ratio.
 12.5.  SUMMARY
      The most  pronounced  effect  of  toluene in animals  is  on  the CN3.   Acute
\exposure to high levels of toluene has been linked with  depression  of the CKS,
 but vapor levels of approximately 1000  ppm appear to have little or no effect on
 gross manifestations  of this  parameter.  A dose-related response of instability,
 incoordination, and  mild  narcosis has been observed in rats  exposed daily to
 toluene vapor at concentrations of  1250 and  1600 ppm, but no effect was noted at
 1100 ppm (Batchelor,  1927).   Inhalation of 1000 ppm toluene  vapor for b hours did
 not increase rearing  reactions  (standing on  hind  legs)  in rats  (Takeuchi  and
 Hisanaga,  1977).    Operand  behavior  (conditioned  avoidance  response)  was
 'unaffected by exposure to 1000 ppm  (Snigeta et  al.  1978)  or  800 ppm (Krivanek
 and Mullin,  1978) toluene.  Inhalation of 1000 ppm for 6 hours/day,  5 days/week
 for 13 weeks did not produce observable behavioral effects  in rats in the pilot
 study for the chronic CUT study  (CUT, I960).  Smyth and Smyth  (1928)  noted tnat
 daily inhalation of 1250 ppm for  ^ hours  each day  for 18 days produced narcosis
 in guinea pigs, while no effect was noted at 1000  ppm during a longer period of
 exposure.   Fabre et al.  (1955)  found that exposure to 2000 ppjs  toluene  for
 8 hours daily, 6 days weekly for  4 months produced only slight nasal and ocular
 irritation  after  transient  initial hyperactivity in one of  two   dogs.    No
 behavioral  effects  were  found  in  rats  and  dogs  after inhalation  of  980 ppm
 "toluene  concentrate"  (150 ppm   toluene)  for  6 hours  daily  for  13 weeks,
 Carpenter et al.  (1976).
      The use of more sensitive methods of detection have,  however,  revealed an
 effect on simple behavioral  parameters  and the CNS  at lower levels.  EEC changes
 were seen  in rats after  inhalation of  1000  ppm  toluene  (Fodor  et al.,  1973;
 Takeuchi and Hisanaga,  1977). A  deficit was noted  in unconditioned reflexes and
 simple behavior at 800 ppm for 4  hours  in rats (Krivanek  and Mullin, 1978), in a
 multiple response schedule  at 571) ppm in rats  (Colotla  et al.,  1979),  and in
 wheel-turning in  rats at  1  ppm  (Horiguchi and  Inoue,   1977).   Neuromodulator
 content in the hypothalamus was affected  at 500 ppm (Andersson et al., 1980).
                                      12-147

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     Early studies suggested a myelotoxic  effect  of toluene.  However, several
studies done since the early 1940's Using  toluene of greater purity have indi-
cated an absence of toluene-induced injurious effect on blood-forming organs in
rats and dogs (von Oettingen et al., 1942a,b; Gerarde, 1959; Wolf et al., 1956;
Fabre et al., 1955; Jenkins et al., 1970; Carpenter et al.,  1976b; CUT, 1980).
Nonetheless,  there is  no  unanimity  on  this point.   Leukocytosis,  impaired
leukopoiesis, and  chromosomal  damage  in the  bone  marrow  have been observed in
some foreign studies (Horiguchi and Inoue, 1977;  Dobrokhotov and Enikeev, 1977;
Lyapkalo, 1973; Yushkevich and Malypheva,  1975).
     Inhalation of concentrations up to  1085 ppn> toluene for 6 weeks or 300 ppm
for 24 months, or ingestion of 590 mg toluene/kg body weight for 6 months, pro-
duced  no liver  damage  (Svirbely  et  al.,  1944; Carpenter et al., 1976b; Jenkins
et  al.,  1970;  CUT,  1980; Wolf et al.,  1956).   Exceptions were the studies of
von Oettingen  et  al.  (1942b),  in  which inhalation  of  600  ppm  toluene caused
increases of  weight and  volume in  the  liver of rats, Fabre et  al.   (1955) in
which  hemorrhagic livers we^e found in dogs, and Ungvary et  al.  (1976) in which
intraperitoneal  injection of 0.43  or  0.82  g/kg  toluene  produced histological
changes in the liver.   However, in  a more recent study by Ungvary et al. (1980),
male CFY rats were exposed  by daily inhalation to  265 ppm or 929 ppm analytical
grade  toluene and female rats were exposed to lower doses five times a week for
up  to 6 months.   No  abnormal  histological  changes  were  found in  the liver,
although growth was inhibited at the higher  dose in males and at the lower dose
in  females;  specific hepatoxic  effects were not noted, although signs of adaptive
compensation were observed.
     Renal  changes consisting  of  casts  in collecting tubules  of  rats  were
observed by  von Oettingen et al.  (1942b)  after inhalation  of 600 ppm toluene.
Hyperemic renal glomeruli and albuainuria were seen in 2 dogs after  inhalation of
toluene  vapors  at concentrations  of  2000 ppm followed  by 2660 ppm  for  4 and
2 months, respectively (Fabre et  al.,  1955).  Slight renal  degeneration has been
observed in guinea pigs  (Smyth and  Smyth,  1928;  Sessa,  1948).  No renal damage
was found  after repeated  inhalation  of 1085 ppm  toluene  for 6 weeks in rats,
guinea pigs, dogs, or monkeys (Jenkins et  al., 1970), after  repeated inhalation
of  300 ppm  for 24 months in rats  (CUT,  1980),  or after repeated ingestion of
590 mg toluene/kg body weight for 6 months in rats  (Wolf et  cJ.., 1956).
     Irritative effects were noted  in the respiratory tract of dogs, guinea pigs,
and rats  following exposure to  toluene vapor (Browning,  1965;  Gerarde, 1959;
                                      12-48

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Fabre  et al.,  1955;  von Oettingen et al., 1942b; Smyth  and Smyth,  1928; Sessa,
1918). Sensitization of  the  heart after  inhalation of toluene has been observed
in mice, rats, and dogs  (Taylor and Harris, 1970; Morvai et al., 1976; Chenoweth,
19*6).
    The acute  oral  LD5Q of toluene is  in the  range of 6.0 to 7.5 g/kg in rats
(Kimura et al.,  1971; Smyth et  al.,  1969b; Withey  and  Hall,  1975;  Wolf et al,,
•1956).  An acute dermal LD^ of 14.1  mg/kg has been  determined  for rabbits (Smyth
et al., 1969b).  Slight to moderate irritation of the rabbit and guinea pig skin
was elicited  by acute or  subacute application of toluene (Kronevi et al., 1979;
Wolf et al.,  1956),  and application to  the  rabbit cornea  has  caused slight to
moderate irritation  (Wolf et al.,  1956; Smyth et al., 1969; Carpenter and Smyth,
1916).
     The inhalation LC   for mice is in the range of 5500 to 7COO  ppm  toluene for
an exposure period of 6 to 7 hours (Svirbely et al., 1943; Bonnet et al., 1979),
An LC  of 8800  ppm of "toluene concentrate"  for 4  hours  (4,038 ppm toluene) was
determined  for  rats  (Carpenter  et  al.,  1976b).    In  guinea  pigs,  inhalation
'exposure to 1000 ppra toluene  for 4  hours caused death in 2 of 3 animals (Smyth
and Smyth,  1928).
     Subchronic treatment of rats  (von Oettingen et al,,  1942b) and rats, guinea
pigs,  dogs, and monkeys  (Jenkins et  al., 1970;  Smyth and Smyth, 1928) at levels
of  200  and  1085 Ppm,   respectively,  did not  have  a  deleterious  effect  on
hematology and  organ pathology.  Horiguchi and Inoue (1977) did report, however,
that mice  showed  changes in blood elements  at  levels as  low  as  10  ppm.   Oral
administration  of  toluene at a level of 590 mg/kg/day for 6 months was tolerated
by rats with  no adverse  effects  (Wolf  et al., 1956).
     The only chronic study of  toluene was the study performed  for CUT (1980) in
which rats were exposed  for  24 months  via inhalation to toluene at levels of up
to 300 ppm.   No effect  on hematology,  clinical chemistry, body weight or histo-
pathology was noted except  for  two  hematologic parameters  in females;  females
exposed to  100 or 300 ppm showed  reduced hematocrit levels  and  increased mean
corpuscular hemoglobin  concentration at  300  ppm toluene.
                                     12-49'

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12.6.  'REFERENCES

ANDERSSON,  K., FUXE, K., TOFTBARD, R., NILSEN, Q.C., ENERGTH, P. and GUSTAFSSON,
J.A.   (1980).   Toluene-induced activation  of certain hypothalamic  and media-
eminence catecholamine  nerve-terminal  systems  of  the male-rat and its effects on
anterior pituitary hormone secretion.   Toxicol.  Letters.   5(6): 393-398.

ANDREWS, L.S.,  LEE, E.W.,  WITHER, C.M.,  KOCSIS, J.J. and SNYDER,  R.   (1977).
   \
Effects  of  toluene on  the  metabolism,  disposition and hematopoietic toxicity of
(3H)benzene.  Biochem.  Pharmaco!.   77(4):  293-300.

BAKER, A.B. and TICHY, F.Y.   (1953).   The effects of the  organic  solvents and
industrial  poisonings  on the central  nervous system.  Proc. Assoc.  for Research
i_n Nervous  and Mental  Disease.  32:  475-505.

BATCHELOR,  J.J.    (1927).    The  relation toxicity  of   benzol  and  its  higher
homologues.  Amer. £.  Hyg.  7: 276-298.

BATTIG,  K.  and  GRANDJEAN,  E.  (1964).   Industrial solvents and avoidance condi-
tioning in  rats.   Arch. Environ. Health.  9: 475-479.

BENIGNUS, V.A.   (198la).  Health effects of toluene:  A review.  Neurotoxicology.
2:  567-58-8.

BENIGNUS, V.A.   (198lb).  Neurobehavioral  effects of  toluene:  A review.  Neuro-
behavioral  Toxicology  and  Teratology.   3:   407-415.

BERGMAN, K.  (1978).   Application of  whole-body autoradiography to distribution
studies  of  organic solvents.  Int. Symp. Control Air Pollut. Work.  Environ. Pt.
2, p.   128-139.

BONNET,  P., RAOULT, G.  and GRADISKI, D.  (1979).  Lethal concentration 50 of main
aromatic hydrocarbons.   Arch  Maladies Prof.,  jte medicine  du  travail et  c[e
Securite Sociale.  40(8-9):  805-810.
                                     12-50

-------
BRAIER,  L.   (1973).   Comparative study of isoc'yclic hydrocarbons ii animals aqd
in man.   Haematologica.   58(7-8): 49U500.

BROWNING, E.,  (1965).  Toxicity and Metabolism of Industrial Solvents.  New York:
Elsevier Publishing  Co.,  p. 66-76.

BRUCKNER,  J.V.  and   PETERSON,  R.G.    (1976).    Evaluation of  toluene  toxicity
utilizing the  mouse as  an  animal model  of human  solvent  abuse.   Pharmacol.
13(2): 244.

BRUCKNER,  J.V.  and PETERSON, R.G.  (1978).  Effect of repeated exposure of mice
and rats to  concentrated  toluene and acetone vapors.  Toxicol. Appl. Pharmacol.
45(1):  359.

BRUCKNER,  J.V.  and  PETERSON, R.G.   (198la).   Evaluation of toluene and acetone
inhalant abuse. I. Pharmacology and phannocodynamics.  Toxicol. Appl. Pharmacol.
£j_: 27-38.

BRUCKNER,  J.V.  and  PETERSON, R.G.   (198lb).   Evaluation of toluene and acetone
inhalant abuse. II.  Model development  and  toxicology.  Toxicol. Appl. Pharmacol.
6J_: 302-312.

CAMERON, G.R.-,  PATERSON,  J.L.H., DE  SARAM, G.S.W.  and THOMAS,  J.C.  (1938).  The
toxicity of  some methyl derivatives  of benzene with special reference to pseudo-
cumene and heavy coal-tar naphtha.  J_, Path. Bact.  46: 95-107.

CARPENTER, C.P.,  SHAFFER,  C.B., WEIL, C.S. and SMYTH,  H.F., Jr.  (1944).  Studies
on the inhalation of 1,3-butadiene; with a comparison of its narcotic effect with
benzol,  toluol,  and styrene,  and a note  on the  elimination  of styrene  by the
human.  J.  Ind.  Hyg. Toxicol.  26:  69-78.

CARPENTER, C.P. and  SMYTH, H.F.   (1946).   Chemical  burns  of  the rabbit cornea.
Amer.  j.  Qpthalmol.  29: 1363-1372.
                                     12-51

-------
CARPENTER,  C.P.,  GEARY,  D.L.,  JR.  and MYERS,  R.C.    (1976a).    Petroleum
hydrocarbon  toxicity studies.   XIII.   Aminal  and human response  to  vapors of
toluene  concentrate.  Topical.  Appl. Pharmacol.  36:  173-190.

CARPENTER,  C.P.,  GEARY,  D.L.,  JR;  and MYERS,  R.C.    (1976b).    Petroleum
hydrocarbon  toxicity studies.  X.  Animal and  human  response  to vapors  of '50
Thinner.1 Toxicol. Appl.  Pharmacol.   36(3):  127-112.

CHEMICAL INDUSTRY INSTITUTE  OF  TOXICOLOGY (CUT).   (1980).   A twenty-four month
inhalation  toxicology study  in  Fischer-311  rats exposed to atmospheric toluene.
Executive  Summary  and  Data   Tables.     Conducted  by  Industrial   Bio-Test
Laboratories,  Inc.,  Decatur, IL, and Experimental  Pathology Laboratories, Inc.,
Raleigh, NC,  for CUT,  Research Triangle Park,  NC.  October 15, 1980.

CHENOWETK,  M.B.   (1916).  Ventricular fibrillation induced  by hydrocarbons and
epinephrine.   J_. Ind. Hyg. Toxicol.   28: 151.

COLOTLA, V.A.,  BAUTISH,  S., LORENZANA-JIMENEZ, M. and  RODRIGUEZ, R.   (1979).
Effects of solvents  on schedule-controlled  behavior.   Neurobehavioral Toxicol.
1 (:):  113-118.

CONTRERAS,  C.M., GONZALEZ-ESTRADA,   T.. and ZARABOZO, D.   (1979).  Petit mal and
grand mal  seizures  produced by toluene or  benzene  intoxication  in  the  cat.
Electroencephalogr.   Clin.  Neurophysiol.  16(3): 290-301.

DELAUNAY, A.,  LEBRUN, J.F.E. and WANG,  H.-S.   (1950).   Action and mechanism of
action of toluene and related compounds on the  permeability of  blood capillaries.
Compt.  Red.  Soc. Biol.  Ill:  58-59.

DIVINCENZO,  G.D. and KRASAVAGE, W.J.  (197D.   Serum ornithine carbamyl trans-
ferase as a liver response  test for exposure  to organic solvents.   Amer.  Ind.
Hyg. Assoc.   j;.   _35:  21-29.
                                     12-52

-------
DOBROKHOTOV,  V.B. and  ENIKEEVj  M.I.   (1975).   Mutagenic  effect pf  benzene,
toluene,  and a  mixture of these  hydrocarbons in  a  chronic experiment.   Gig.
Sanit.   _^:  32-34.   (In Russian with English. summary;  evaluation based on  an
English translation  provided  by  the  U.S.  EPA).

ELOVAARA,  E., HEMMINKI,  K.  and VAINIO,  H.   (1979).   Effects  of  methylene
chloride,  trichloroethane, trichloroethylene,  tetrachloroethylene and toluene on
the development  of chick embryos.  Toxicology.  12(2):  111-119.

FABRE,  R.  et al.    (1955).    Recherches  toxicalogiques  sur  les solvents  de
remplacement due benzenede.   Archives Maladies  Professionalles je Medicine  d_u
Travail et de Securite  Sociale.   16:  197-215.   (Cited in Bergman,  1979).

FAUSTOV,   A.S.    (1958).   Toxicity   of  aromatic  hydrocarbons.  I.  Comparative
tbxicity of some aromatic  hydrocarbons.  II. Some problems of the toxic-hygienic
properties of aromatic  hydrocarbons.  Trudy Voronezh. Med.  Inst.   35:  247-255,
257-262.

FODOR,  G.G., SCHLIPKOETER, H.W. and  ZIMMERMANN, M.  (1973).  The Objective Study
of Sleeping Behavior in Animals  as a Test  of  Behavioral Toxicity.   In:  Adverse
Effects of Environmental Chemicals and Psychctropic Drugs.   Quantitative Inter-
pretation of Functional Tests. Gernany, Elsevier Science Publishing Co.,  Vol.  1,
p. 115-123.

GELLER, I.,  HARTMANN, R.J.,  RANDLE,  S.R.  and CAUSE, E.M.   (1979).   Effects  of
acetone and toluene  vapors on  multiple  schedule  performance of rats.   Pharm.
Biochem.  Behavior.   11: 359-399.

GERARD'E,  H.W.  (1959).   Toxicologi.cal studies on  hydrocarbons.  III.  Arch.  Ind.
Health.  19: 403-418.

GRADSKI,  D., BONNET,  P., DUPRAT, P.,  ZISSU, D., MAGADUR, J.L., and GUENIER,  J,P.
(1981).    Etude  toxicologique  chronique  par   inhalation chez   le  rat  de
I'association benzene-toluene.   Toxicol.  Europ.  Res.   3:  201-206.
                                     12-53

-------
GUSEV, I.S.  (1967).  Comparative toxicity of  benzene, toluene and xylene.  Biol,
Deistvie Gig.  Znachemie Atmos.  Zagryaznenii.  10: 96-108.   Taken  from:   Chem.
Abst. 69:17711e,  1967.

HIROKAWA,  T.   (1955).   Studies on the  poisoning  by  benzol  and its homologues.
III.  Experimental studies on the sexual differences of blood picture.  Jap. J_.
Med. Sci.   Biol.   8:  279-281.

HORIGUCHI, S.  and  INOUE,  K.   (1977).   Effect?  of toluene on the wheel-turning
activity  and  peripheral  blood findings  in  mice - an  approach to  the maximum
allowable concentration of toluene.  J_. Toxicol. Sci_.  2CQ: 363-372.

HUDAK,  A.,  BORS,  Z., UNGVARY,  G. and  TOLLY,  G.    (1976).    Reversibility and
interaction with  hepatic  regeneration  of toluene reduced liver injury.   Acta
Morphol. Acad. Sci.  Hung.  21(1-2):  153-166.

IKEDA,  M.  and OHTSUJI,  H.   (1971).    Phenobarbltal-induced  protection against
toxicity of toluene and benzene in the  rat.   Toxicol.  Appl.  Pharmacol.  20( 1):
30-43.

IKEDA, T. and MIYAKE, H.   (1978).   Decreased  learning in rats following repeated
exposure to toluene:  Preliminary report.  Toxicol. Lett.  1 (1Q: 235-239.

IKEDA, I., KAEHARA, N.,  SADMOTO, T.,  HARABUCHI, I., YAMAMURA, K. and MIYAKE, H.
(1981).   Effects  of toluene  exposure  on   the  rest-activity  cycle  of  rats.
Toxicol. Lett.  9(3_):  255-266.  Taken  from Chem.  Abstr. 95_:2l6001k,  1981.

ISHIKAWA, T.T. and SCHMIDT,  'I.,  Jr.   (1973).  Forced turning induced by toluene.
Pharmacol.  Biochem.  Behav.  1(5): 593-595.

JENKINS, L.J., Jr.,  JONES, R.A.  and  SIEGEL,  J.   (1970).   Long-term  inhalation
screening  studies  of benzene,  toluene,  o-xylene,  and cumene  on  experimental
animals.  Toxicol.  AppK Pharmacol.   16: 818-823.
                                     12-51

-------
KEPLINGER, M.L., LANIER, G.E. and DEICHMANN, W.B.  (1959).  Effects of environ-,
mental  temperature  on  the  acute toxicity  of a  nuniber  of compounds  in rats.
Toxicol. Appl.  Pharmacpl.  J_: 156-161.

KHINKOVA, L.  (1974).   Experimental  data on the toxicity of some organic solvents
used in the furniture industry.  Tr. Inst Khig,  Okhr.  Tr. Prof.  Zabol.  22(1):
                                  _    g—                        _—
133-140.  Taken from:  £hera. Abst.  _88:11?0j,  1978.

KIMURA, E.T., EBERT, P.M. and DODGE,  P.W.  (T97D.  Acute toxicity and limits of
solvent residue for sixteen organic  solvents.   Toxicol. Appl. Pharmacol.  19(4):
699-7014.  Taken from:  Chem. Abst. 75: "39139u,  1971.

KOGA,  H.  and OHMIYA,  Y.   (1978).   Potentiation  of  toluene  toxcity by hepatic
enzyme  inhibition in mice.  J_. Toxicol. Sci.   3(1): 25-29.

KOJIMA, T. and KOBAYASHI, H.   (1975).  Toxicological study on toluene poisoning
by  inhalation.   Toluene poisoning in  the  hypoxic atmosphere.   Nippon Hoigaku
Zasshi.  29(2): 62-87.   (Cited in NRC, 1980).

KRIVANEK, N. and MULLIN, L.S.  (1978).  Comparison of conditioned avoidance and
unconditioned  reflex  tests in rats  exposed by  inhalation to  carbon monoxide,
 1,1,1-trichloroethane,  toluene,  ethanol.   Toxicol.  Appl.  Pharmaeol.   45(1):
357-358.

KRONEVI,  T.,  WAHLBERG, J.  and HOLMBERG,  B.   (1979).  'Histopathology of skin,
liver,  and  kidney after epicutaneous administration of five industrial solvents
to  guinea pigs.  Environ. Res.   19(1): 56-69.

LYAPKALO, A.A.  (1973).  Genetic activity of benzene and toluene.  Gig. Tr. Prof.
Azbol.   V7: 24-28.   (In Russian with English summary;  evaluation  based on an
English translation provided  by  the U.S. EPA).

MATSUMOTO,  T.,  TAKEUCHI, Y.,  TANAKA,  T.  and  MAEDA, K.   (1971).  Experimental
studies on the chronic toluene poisoning. 3. Effects of  toluene exposure on blood
and  organs  in  the rats.  Sangyo  Igaku.  Jap. «J.  Indust. Health.  13; 501-506.
                                      12-55

-------
MORVAI, V., HUDAK, Ai and VARGA, U-B.   (1976).  ECG changes in benzene, toluene,
and xylene poisoned*  Acta Med. Acad.  SCi. Hung.   33(3): 275-286.

MULLIN, L.S. and KRIVANEK, N.D.  (1982).   Comparison of unconditioned reflex and
conditioned avoidance  tests  in rats exposed by inhalation to  carbon monoxide,
1,1,1-trichloroethane, toluene, or ethanol.  Neurotoxicology.  3:  126-137.

NIOSH (NATIONAL INSTITUTE  FOR OCCUPATIONAL SAFETY AND HEALTH).  (1973).  Criteria
for a  Recommended Standard.   Occupational Exposure to  Toluene.   Final Report.
Contract No. HSM-99-72-118.  Available .through NTIS,  NTIS No. PB-222-219/8, 108
P.

NRC  (NATIONAL  RESEARCH COUNCIL).   (1980).   The Alkyl Benzenes.   Committee on
Alkyl Benzene Derivatives, Board on  Toxicology and Environmental Health Hazards;
Assembly of Life  Sciences, National Research Council.   Washington, DC: National
Academy Press.

PETERSON,  R.G.  and BRUCKNE.i,  J.V.   (1978).  Measurement of  toluene levels in
animal tissues.   In:  Voluntary Inhalations of Industrial Solvents.  C.W. Sharp
and Carrol, L.T., eds.  Rockville, MD: Nat. Inst. Drug Abuse.  24: 33-^2.

POWERS, M.B.  (1979).  Memorandum for  the  Record from the NTP Chemical Selection
Group, Toxicology Branch,  CGT,  DCCP, National Institute, Washington, DC, May 25,
 1979.

PRYOR, G.T., BINGHAM, L.R. and  HOWD, R.A.  (1978).  Behavioral toxicology in rats
of a mixture  of solvents  containing substances subject to inhalation abuse by
humans.  Toxicol. Appl. Pharmacol.  *I5( 1): 252.

REYNOLDS,  E.S.   (1972).    Comparison  of early  injury  to  liver  endoplasmic
reticulum  by halomethanes,  hexachloroethane,  benzene,  toluene,  bromobenzene,
ethionine, thioacetamide, and  dimethylnitrosamine.  Biochem.  Pharmacol.  21(9):
2555-2261.

SAVOLAINEN, H.   (1978).   Distribution and nervous system binding of intraperi-
toneally injected toluene.  Acta Pharmacol. Toxicol.  ^3(1):  78-80.
                                      12-56

-------
SCHOLZ, R., SCHMITZ, H., BUCHER, T. and LAMPEN, J.O.  (1959).  Effect of nystatin
on yeast.  Blochem.  331; 72-86.

SCHUTZ, E.  (1960), Effects on organic liquids on the skim Arzneimittel-Forsch.
_10: 1027-1029.

SESSA, T.   (1948).  Histopathology in experimental chronic  toluene  poisoning.
Folia Med.  (Naples).  3±: 91-105.  Taken from:  Chem. Aust." 42:  I666b,  1948.
SHIGETA, S.f  AIKAWA,  H., MISAWA, T. and  KONDO,  A.   (1978).  Effect  of  single
exposure to toluene on Sidam avoidance response in rats.  J_. Toxicol. Sci.   3(4):
305-312.
SLIMAK,  M.    (I960).   Exposure Assessments  of Priority Pollutants:   Toluene.
Report (draft) prepared by Arthur D. Little,  Inc., MA.  Prepared for U.S.  Envir-
onmental  Protection  Agency,  Monitoring and Data Support  Division,  Washington,
DC.

SMYTH, H.F., JR., WEIL, C.S., WEST, J.S.  and CARPENTER, C.P.  (1969a).  Explora-
tion of  joint toxic action: Twenty-seven industrial chemicals intubated in  rats
in all possible pairs.  Toxicol. Appl. Pharmacol.   14(2):  340-347.

SMYTH, H.F., JR., CARPENTER,  C.P.,  WEIL, C.S., POZZANI, U.C., STRIEGEL,  J.A.  and
NYCUM,. J.S.  (1969b).  Range-finding toxicity data. VII.  Amer.  Ind.  Hyg.  Assoc.
J.  30(5): 470-476.
SMYTH, H.F. and SMYTH,. H.F.,  JR.   (1928).   Inhalation experiments with certain
lacquer solvents.  J_. Ind. Hyg.  10: 261-271.

SPECK, B. and MOESCHLIN, S.   (1968).  Effect of toluene, xylene, chloramphenicol,
and thiouracil on bone marrow.  Experimental autoradiographic study with thymi-
dine-3hh  Schweiz. Med. Wochenschr.  98(42): 1683-1686.
                                     12-57

-------
SVIRBELY, J.L., DUNN, R.C. and VON OETTINGEN, W.F.  (1943).  The acute toxicity
of vapors  of certain  solvents containing  appreciable  amounts of  benzene and
toluene.  J^.  Ind. Hyg. Toxicol.  25: 366-37 J.  Taken from:  Chem. Abst.  39.: 4979,
1945.

SVIRBELY, J.L., DUNN, R.C. and VON OETTINGEN, W.F.  (1944).  J. Ind. Hy_g_.  26_:
37-46.  Taken from: Chem. Abst. 38_: 4696, 1944.

TAHTI, H., RUUSKA,  J.  and VAPAATALO, H.   (1977).   Toluene toxicity studies on
rats after "i week inhalation exposure.  Acta  Pharmacol.  4 1 (*t): 78.

TAKEUCHI, Y.  (1969).  Experimental studies  on the  toluene  poisoning—chiefly on
the findings of peripheral blocd and adrenal  gland.  Ind. Health^  ]_: 31-45.

TAKEUCHI, Y., T. TANAKA,  T.  MATSUMOTO and T. MATSUSHITA.   (1972).  Response of
dience-phalon-hypophysis  adrenal  cortex system  in exposure to  toluene  vapor.
Sangyo Igakus 14(6): 543-553.
I

TAKEUCHI, Y. and HISANAGE,  N.   (1977).    The  neurotoxicity of  toluene:   EEC
changes  in   rats exposed to  various concentrations.   Brit.  J^.  Med.   34(4);
314-324.

TAKEUCHI,.Y. and SUZUKI, H.  (1975).  Change  of  convulsion  threshold of the rat
exposed to toluene.  Indust. Health.  13: 109-114.

TAKEUCHI, Y.; ONON, Y.; and HISANAGA, N.  (1981).  An experimental study on the
combined  effects of n-hexane and  toluene on  the peripheral nerve of  the rat.
Brit. J_.  Indus.  Med.  _3_8:  14-19.

TAYLOR, D.C. and HARRIS, W.S.  (1970).  Glue sniffing causes heart  block in mice.
Science.  1?0: 866-868.
   !        ~ ~ L

TSUZI, K.   (1956).   Convulsion caused  by phenol compounds.  Kumamoto Med.  j):
152-164.  Taken from: Chem. Abst.  51: 9909g,  1957-
                                     12-58

-------
UNGVARY, G.,  HUDAK, A., BORS, Z. and FOLLY, G.   (1976).  The effect of toluene on
the liver assayed by quantitative and morphological methods.   Exp. Mol. Pathoi.
25(1):  49-59.
UNGVARY, G.,  HAMORI,  J.  and  HUDAK, A.   (1975).   [Experimental study  of  the
hepatotoxic  effect  of toluol.  II.  Electron  microscopic  and electron  histoc-
hemical studies.]  Morphol.  Igazsagugyi Ory. Sz.  15(4):  256-263.

UNGVARY, G., MANYAI, S., TATRAI, E.  (1980).   Effects of toluene inhaltion on the
liver of rats — dependence on sex,  dose and exposure  time.  _J. f^y_g. Epid. Micr.
Immun.  24_:  242-252.

U.S. EPA (U.S.  ENVIRONMENTAL  PROTECTION  AGENCY).   (1980a).   Priority Pollutant
Frequency Listing Tabulations  and Descriptive Statistics.  Memo from D. Neptune,
Analytical  Programs  to  R.B.   Shaffer,  Director  of  Effluent Guidelines  Div.,
January, 1980.  (Cited in Slimak, 1980).

U.S. EPA  (U.S.  ENVIRONMENTAL   PROTECTION  AGENCY).   (1980b).  Volatile  Organic
Compound (VOC)  Species Data Manual,  2nd  ed.,  Publication  No.  EPA-450/4-80-01D.
Office  of  Air,  Noise,  and  Radiation,   Office  of  Air  Quality Planning  and
Standards, Research Triangle Park, NC.

VON  OETTINGEN,  W.F.,  NEAL,  P.A. and DONAHUE, D.D.   (1942a).  The toxicity ar.d
potential dangers of  toluene—Preliminary report.   J^. Amer. Med. Assoc.   118:
579-584.

VON   OETTINGEN,   W.F.,   NEAL,   P.A.,   DONAHUE,   D.D.,    SVIRBELY,    J.L.,
BAERNSTEIN, H.D., MONACO, A.R., VALAER,  P.J.  and  MITCHELL,  J.L.   (1942b).   The
Toxicity and Potential Dangers of Toluene with Special Reference to its  Maximal
Permissible Concentration.  U.S. Public Health Serv.  Puh.  Health Bull. No. 279,
50 p.

WAHLBERG,  J.E,    (1976).    Percutaneous  toxicity  of  solvents.    A  comparative
investigation  in  the  guinea   pig with  benzene,  toluene,  and  1,1,2-trichloro
ethane.  Ann. Occup. Hyg.   19(2):  115-119.  Taken from: Chem. Abst.  86^:  66415W,
1977,
                                     12-59

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WITHEY, R.J, and  HALL,  J.WV  (1975).   Joint  toxic action of perchloroethylene
with benzene or toluene in rats.  Toxicology.   Ml): 5-15.

WOLF, M.A. et al.  (1956).  lexicological studies of certain alkylated benzenes
and benzene.  Arch. Ind. Health.  14: 387.

YAMAMURA, K.,  IKEDA,  T., MAEHARA, N.,  SADMOTO,  T. and HAKABUCHI,  I.   (1981).
Effects of toluene exposure on blood pressure and its responsiveness to impulse
noise in rats.  Toxicol. Lett.  9(1Q:  361-366.  Taken from Chera. Abst.  96: l686x,
1982.

YAMAWAKI, S.,  SEGAWA,  T. and SARAI,  K.   1982.   Effects of acute  and chronic
toluene inhalation on  behavior  and  (3H)-serotonin binding in rats.   Life Sci.
30:  1997-2002.

YUSHKEVICH, L.B.  and MALIPHEVA,  M.V.   (1975).   Study  of  the bone marrow as ah
index  of experimentallyinduced  poisoning  with  chemical  substances   (such  as
benzene and its homologs).  Sanit. Tokslkol. Metody Issled. Gig.   36.   (Cited in
U.S. EPA, 1980).
                                     12-60

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         13.  PHARMACOKINETIC CONSIDERATIONS IN HUMANS AND IN ANIMALS

13.1.   ROUTES OF EXPOSURE .AMD ABSORPTION
    For hmnans, the most common routes  of  exposure  to toluene are through the
respiratory tract and the skin.  Toluene is absorbed readily through the respira-
tory tract.  In  experimental  exposures  of humans to toluene conducted by Astrand
and coworkers (1972; also reported  in  Astrand, 1975), toluene was  detected in
 \
arterial blood during the first  10 seconds of exposure.  Toluene was supplied in
the inspired air 'at  100  or  200 ppm through  a  breathing valve and  mouthpiece.
Unless otherwise specified,  in the  experiments  reported here,  human  subjects
breathed toluene vapor  from  some type of respiratory  apparatus.    In  resting
subjects,  the  concentration  of toluene in arterial  blood increased  rapidly
during the  first 10  minutes of exposure and then began to level  off,  approaching
an apparent steady state  by 30 minutes.  The concentration of toluene in alveolar
air (i.e.;  an  air  sample taken at  the  end  of a normal expiration)  increased
concoinitantly.
    Alveolar and arterial  concentrations  of  toluene were  proportional  to the
concentration in inspired air.  At the  end of  30 minutes of  exposure to 100 or
200 ppm (0.375 or 0.750 mg/2,) toluene,  the concentration of toluene in alveolar
air (mg/X,) was  18$ of that in  inspired air (mg/R,), while  the  concentration in
arterial blood (mg/kg) was 270$. of that in inspired air (mg/£)  (Astrand et al.,
 1972;  Astrand,  1975).  The ratio between  arterial blood ana alveolar air concen-
trations was  15, which  is similar to  the _in vitro  blood/air partition coeffi-
cients (at  37°C) of  14.6,  15.6,  and 15.6  reported for human blood  by Sato et al.
 (197^a), She>"wood (1976), and Sato and Nakajima (1979a), respectively.
     According to Veulemans and Niasschelein  (1978a),  subjects' lung clearances
 (i.e., the  virtual  volume of inspired air from which all  available toluene is
absorbed per unit time) decreased during exposure at rest,  reaching an apparent
steady state 9 to 13 minutes  from the  beginning  of  exposure.   Lung clearance
 (C.-C  )/C.  x V   where  C.  is the  concentration of toluene in inspired air (mg/JZ,),
  1616        1
C  is  the concentration of toluene in expired air  (mg/2,), and V  is the respira-
 C                                                            6
tory minute volume (fc/min).  Lung clearance  varied  less among  individuals than
did the concentration in expired air.
    Nomiyama and Nomiyama (197^a) measured the pulmonary retention  ((C.-C )/C.
x 100) of volunteers exposed  to  about 115 ppm toluene for H hours.  The subjects
                                      13-1

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may have  been  fairly sedentary because  the  authors did  not  mention exercise.
Retention at the end of 1 hour was approximately 52? and decreased to 37$ at the
end of  2  hours,  remaining constant  at that  "".evel  for the  remaining  2 hours.
These  results  suggest  a  slower  approach  to  steady-state  concentrations  in
expired or alveolar air than  was indicated by  the time courses obtained for lung
clearance by Veulemans  and Masschelein (1978a) or  for  alveolar air concentra-
tions by Astrand et al. (1972).  The results  also suggest a lower percentage of
uptake or retention  than was reported  by Veulemans and Masschelein  (1978a) and
others as will be  presented  subsequently.  The reasons for these discrepancies
are unclear.
     Eyercise affected the absorption of toluene through the respiratory tract.
In the  experiments of Astrand  and coworkers  (Astrand  et al.,  1972;  Astrand,
1975),  exercise  greatly increased the  concentrations  of toluene  in  arterial
blood and alveolar air of the subjr-c,ts  during  exposure,  and these concentrations
did not level  off as soon in  exercising  subjects  as in resting subjects.   The
concentrations of  toluene in arterial blood and alveolar air were approximately
the same  at 30 minutes of exposure  to  200 ppm during rest as at 30 minutes of
exposure to 100 ppm during  light exercise (50  watts). At 30 minutes  exposure to
100 or 200  ppm (0.375  or 0.750 mg/X,) toluene, the concentrations in milligrams
per liter expressed relative  to the concentration in insplrad air  (mg/fc) were 33?
for alveolar air and 620? for arter.lal  blood at exercise of 50  watts,  and 47? for
alveolar air and 725? for arterial blood at exercise  of  150 watts.  The ratio of
arterial  to alveolar concentration remained  about  the  same  as at rest.   Thus,
alveolar  concentrations appeared to  reflect  arterial concentrations  during
exposure to 100 to 200 ppz toluene at rest and various  intensities of exercise.
     The  inhalation  of  4?  C0? by resting subjects  during exposure  to  100 ppm
toluene increased  their  alveolar  ventilation  (£/min) and the concentrations of
toluene in  their arterial  blood  and  alveolar  air (Astrand et  al.,  1972).   The
increased toluene  concentration in blood and  alveolar air were similar to those
obtained with a corresponding increase in alveolar ventilation during exercise.
Because exercise  increased both alveolar ventilation and  heart  rate while CQ
increased only alveolar ventilation, the  effect  of exercise on toluene absorp-
tion appears to be due to increased alveolar  (or pulmonary) ventilation.
     In the experiments of  Veulemans  and Masschelein (1978a), the "steady state"
lung clearances of 6 different subjects  during exposure to 50  ppm toluene at rest
and  at  workloads  of  25 and  50 watts  on a  bicycle ergometer  correlated  well
                                      13-2

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  o
(r  = 0.96) with their  respiratory  minute volumes.  Lung  clearance  was deter-
                              i
mined from  the regression  line to be  equal  to 0.47 tf  .   The uptake  rate  in
milligrams  per  minute,  which  equals  lung  clearance  times   the   inhaled
concentration, therefore was equal to 0.47 tf C. (where C. is expressed in mg/£)
                                            61         X
and total uptake in milligrams  equaled  47$  of the total amount inhaled.   Lung
clearances and respiratory minute volumes doubled with an exercise intensity of
25 watts  and  tripled  with  an  exercise  intensity   of   50 watts   over  the
corresponding values at rest (Veulemans and Masschelein, 1978a).
     Carlsson and Lindqvist (1977)  found  that the uptake of  toluene  by 7 male
subjects exposed to 100 ppm for 30  minutes  (0.375 mg/Jl)  during rest  or various
levels of exercise  (50,  100,  and 150 watts on  a  bicycle ergometer)  correlated
inversely (r  = 0.72) with  the  alveolar concentration  determined  at  the end of
30 minutes exposure, as described by the following equation:
              1 U tak   -   n  6^ alveolar  concentration (mg/fc)  x 100    + 72.9
                                 inspired concentration  (mg/L)
This  relationship  is  logical and applies to other  solvents  as well (Astrand,
1975; Ovrum et  al., 1978).   Percent uptake  was determined on  the  basis of the
total amount  of toluene inhaled and exhaled  during  the entire exposure period
(i.e., the expired air was collected continuously throughout exposure, and thus
was a mean value).   The uptake ranged from about 47 to 67$ at rest and from about
36 to 57$ at an exercise level of 150 watts.  This  group  of men  comprised 3 thin,
1  slightly  overweight, and  3   cbese subjects (Carlsson and  Lihdquist, 1977).
Similar uptake values were also more recently reported by Carlsson (1982) for a
group of  12 subjects who were exposed to 80 ppm toluene  during rest (=50$), and
during a fourth consecutive 30-minute period  of 150W exercise  (=30$).
     Cvrum and coworkers (1978), monitoring four workers exposed to toluene in a
printing  plant, found  good agreement  between  the  value  for percent uptake
determined directly from the total amounts of toluene inspired and expired during
a  sampling  period  and the  value determined  indirectly  from  the   instantaneous
concentrations  in  alveolar  and  inspired  air, using the equation  given in the
preceding paragraph.  Percent uptake determined by the direct method was 47$ and
by the  indirect method was 51$.  The  total  uptake of  toluene that would occur
during exposure to 80  ppm (0.3  mg/£.) for an 8-hour work day was calculated using
the mean value for  pulmonary ventilation of 16 H/min measured for these 4 workers
and a percent uptake of 50.  The total  uptake amounted to approximately 1150 mg
(Ovrum et al.,  1978).
                                      13-3

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 \
     The percent  uptake values  determined by  Carlsson and  Lihdquist  (1977),
Ovrum et al.  (1978) and Carlsson  (1982) are in  reasonable  agreement with those
previously reported in abstracts from the foreign literature:   51)? average uptake
during 5 hours'  exposure to 271 to 1177  Hg/£ (Srbova and Teisinger, 1952) and 72/6
initial  retention  decreasing to  57$  retention  towards  the end of  8 hours of
exposure to 100 to 800 ng/S, (Piotrowski, 1967).
     Another factor, in addition  to exercise,  that  has  been reported to affect
the absorption of  toluene through  the respiratory tract is the amount of adipose
tissue in the body (Carlsson  and Lindquist,  1977; Carlsscn and Lindquist, 1982).
Carlsson and Lindquist (1977) found that mean  alveolar  air concentrations were
slightly higher in  3  thin  men than in 3 obese  men at the  end of 30 minutes of
exposure to 100 ppm (0.375 mg/K.)  toluene during rest or exercise.   The ranges,
however, overlapped.  Conversely,  the total uptake of toluene duri.ig 30 minutes
of exposure (determined as  previously described) was lower for the thin subjects
than for the obese ones (Table 13-1).

                                   TABLE  13-1
   f
            Uptake of Toluene in Thin and Obese Men Turing Exposure
              to a Toluene  Concentration of 375  mg/m   (100  ppm)a'

Number of
Subjects

Thin (N = 3)
Mean
Range
Slightly overweight
(N = 1)
Obese (N = 3)
Mean
Range

Adipose
Tissue
(kg)

6.0
1.4-10.7

22.8

44.0
35.1-49.0


Rest


61
55-69

71

84
72-73
Uptake

50 W


148
133-158

179

198
183-206
(mg)
Exercise
100 W


193
168-211

246

258
237-275


150 W


228
181-271

299

319
258-358
 Source:  Carlson and Lindquist, 1977
 b
 The subjects were exposed during one 30-minute period of rest and three
 consecutive 30-minute periods of exercise in order of increasing intensity.
 A 20-minute pause without exposure occurred between rest and exercise.
 Expired air was collected continuously during exposure.

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The thin subjects had a mean adipose  tissue  content  of 6 kg and the obese ones
had a mean  adipose tissue content of  M  kgr   It appears,  from Figure 6 in the
Carlsson and Lindquist (1977)  paper,  that  the obese men inspired a greater total
quantity of toluene than did the thin men.  Because the concentrations of toluene
in the inspired  air  were  the  same for both  thin and obese subjects,  pulmonary
ventilation must have  been  greater  in the obese ones.  Thus the differences in
uptake between the thin and obese men may have  been  at  least  partially due to
greater  ventilation  (respiratory minute  volume) in  the  obese subjects rather
than to their adipose tissue per se.  Veulemans and Masschelein (1978a) reported
finding no correlation between  a  subject's content of adipose tissue and uptake
of toluene  during exposures to 50  to  150 ppm toluene  lasting  about  4.hours.
Astrand  and  coworkers  (1972)  stated that  they  found no systematic differences
between  male  subjects  (N - 11,  adipose  tissue 5.7 + 1.5 kg, mean  + S.D.)  and
female subjects  (N = ^, adipose  tissue 13-3 kg, mean; 9-6 to 20.2 kg,  range) in
alveolar air and arterial blood  concentrations of toluene.
     Dahlmann  and  coworkers   (1968a,   1968b)  investigated  the absorption of
toluene contained in cigarette  smoke through the mouths and  respiratory tracts of
volunteers.  The uptake of toluene from smoke that stayed in the subject's mouth
for  2 seconds  or less  and was  not  inhaled was ?9J;  uptake  when the  smoke was
inhaled  into the lungs was 93$.   It is unclear whether each  subject was exposed
to a single  puff of  smoke,  the smoke from  1  cigarette  (8  puffs),  or  the smoke
from 2 cigarettes.
     During  inhalation  exposure of   resting subjects,  the  concentration of
toluene  in peripheral venous blood  (from  the cubital vein of the arm)  attained
apparent steady  state  more  slowly than did lung clearance or concentrations in
alveolar air or arterial blood  and was more variable among subjects than were the
above mentioned values (Veulemans and Masschelein, 1978a;  1978b; Astrand et al.,
1972; Sato  and Nakajima,  1978).  Peripheral  venous concentrations  appeared to
level off  during the second or third  hour of exposure.  Von Oettingen (19^2a,
19^2b) had observed  that  toluene concentrations in subjects' peripheral venous
blood  at  the  end  of  8  hours  of  exposure were  roughly  proportional  to the
concentrations of  toluene (200  to  800 ppm)  in the  atmosphere  of  the  exposure
chamber.
     Similar results were obtained when the toluene concentrations in peripheral
venous blood of 19 workers at the  end of a work week v:ere  correlated with toluene
concentration in workplace air; the data points showed considerable scatter, but
                                      13-5

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a positive correlation (rc  =  0.78) was observed.  (Apostoli et  al.^  1982).  IJlood
was sampled at the end of the work shift Friday afternoon and air was sampled for
20-25 minutes with personal sampling devices once during the same afternoon.  The
concentrations  of toluene in  venous  blood ranged  from 3^ to  572 ng/&  and in
workroom  air  from  15  to  182 \ig/i  (57-686 ppm).    The  ratio  of  toluene
concentrations in peripheral venous blood  (ng/£.)  to that in air (|ig/Z) was =3.
The  authors  calculated  similar  yalues  from  the  data  of  otherp  for  both
experimental  and  occupational  human exposure  (Astrand  et al.,   1972;  Veulemans
and Masschelein,  1978b; Ovrum et al.,  1978; Angerer and Behling, 1981).
     Veulemans   and  Masschelein   (1978b)   reported   that   the  steady-state
concentrations of  toluene  in peripheral  venous blood were correlated with the
rate  of uptake  at  different  inspired  concentrations  (50,   100,  and 150 ppm)
(r2 = 0.73) arid at different levels of rest and  exercise (r2 _  0.74).  In both
instances, the relationship  between peripheral venous concentrations and uptake
rate was:
     Venous concentration  (mg/2,) = 0.3 min/X, x uptake rate (mg/min).
The concentration of toluene in peripheral  venous blood of exercising subjects
    i
increased more rapidly and appeared to reach steady-state  values sooner than in
resting subjects  (Astrand et al.,  1972;  Veulemans and Masschelein, 19?8b).
     Absorption through the  respiratory tract has been  studied  less extensively
in experimental animals than in humans.   The initial uptake of  a relatively low
concentration  of  toluene  was  found to  be  approximately  90% in dogs inhaling
toluene  (Egle and Gochberg,  1976).   Varying  the  ventilatory  rate  from  5  to
HO inhalations per minute, the  tidal volume from 100 to 250 m£, or the concentra-
tion of  toluene  from 0.37 to 0.82 ^g/i  (approximately  100 to  220 ppm)  had no
significant  effect  on  the  animals'  initial respiratory uptake.  Toluene was
readily absorbed from the upper as well as from  the lower respiratory tract.  The
dogs  were anesthetized  with  sodium  pentobarbital  for  these  experiments and
breathed toluene from a recording respirometer for  1 to 2 minutes.  The percent
uptake  was  calculated  from  the  total amounts  of  toluene  inhaled and exhaled
during the 1 to 2 minute exposure.
     von Oettlngen and coworkers  (1942b)  found  that  the  concentration of toluene
in the  peripheral  venous blood of dogs  at  the end of  8  hours  of  exposure was
proportion.il to the concentration  of toluene  (200, 400,  or 600 ppm) in  the air of
the exposure  chamber.   As previously described,  similar observations had been
made with humans.
                                      13-6

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     Mice exposed  singly  to an extremely high initial concentration of methyl-
  C-toluene. in a closed chamber for 10 minutes retained about 60? of the radio-
activity  when removed from the chamber  at  the end  of  the exposure  (Bergman,
1979).  This  value is a  rough approximation of  absorption  because  some of the
toluene may have been adsorbed to the  animals'  fur.  A substantial portion of the
retained dose appears to  have been absorbed,  however,  as shown by its subsequent
excretion in  the urine (Section 13-4.).  The initial concentration of toluene in
the chamber (10 [ii  evaporated  in  a volume of about 30 m&, or about 77,000 ppm)
would  have  been  above the saturation concentration even if the temperature had
been   as  high   as  30°C   (saturation  concentration  =  48,900 ppm  at  30°C)
(Verschueren, 1977).  Bergman  (1979)  noted that  exposure to toluene under these
conditions markedly reduced the respiratory rate of the  mice and attributed this
reduction to  irritation.   It  seems more likely that the decreased respiratory
rate was due  to narcosis.
     Absorption of toluene also occurs through the skin.  Dutkiewicz and Tyras
(1968a,  1968b),  in experiments with  humans, measured the absorption of liquid
.toluene into  the skin of  the forearm and found  the  rate of absorption to be  14 to
,23 mg/cm /hr.  This rate was calculated from the difference between the amount of
toluene  introduced  under  a watch glass affixed  to the  skin and  the amount
remaining on the skin at the end of 10 to 15 minutes.  Absorption  of toluene from
aqueous solutions  during immersion of both hands  was  160 to 600  [ig/cra  /hr and was
directly proportional to the initial concentration of toluene (180 to 600 mg/ii).
From  these  results,  Dutkiewicz and  Tyras  (1968a,  1968b)  calculated  that the
absorption  of toluene through the skin of both  hands  during contact with a
saturated aqueous  solution of  toluene for 1 hour could be in the same  range as
absorption  through the respiratory tract during  8  hours of exposure to  26.5 ppm
 CO. 1 mg/JL)  toluene.
     Sato and Nakajima (1978)  found,  however,  that the maximum toluene concen-
tration  (170  |ig/£) in the blood of  subjects  who  immersed one  hand  in liquid
toluene for 30 minutes was only 26% of  the  concentration  (650 ng/2.) in  blood of
subjects who  inhaled 100  ppm toluene  vapor  for 30  minutes.  Blood was collected
from  the cubital  vein of the (unexposed)  arm  at intervals  during  and after
exposure.   Sato and Nakajima  (1978) suggested  that some of  the  toluene that
penetrates  the stratum corneum may be subsequently  given off into the  air, rather
than entering the systemic circulation.  Toluene appears to pass slowly  from the
skin into the bloodstream after penetrating the skin.  Guillemin et al. (1974)
                                      13-7

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reported that  the  elimination of toluene  in alveolar  air  sometimes increased
during the first 20 minutes after the  termination of exposure of both hands to
liquid toluene, and Sato and  Nakajima  (197?) noted that  the  maximum levels of
toluene in venous  blood were  maintained for  about  15 minutes after the end of
exposure.
     Jakobson  et  al,  (1982)  monitored  the  concentration  of  toluene  in  the
arterial blood of anesthetized guinea pigs following epicutaneous exposure.  In
                     2
this  study,  a 3-1 cm   area of clipped  back skin was  continuously exposed to
liquid toluene by means of a sealed glass ring.   It  was found that the concen-
tration of  toluene in  the  blood  increased  rapidly within  1  hour  to. a peak of
-1.3 Jig/mi, and then decreased in spite of the continuing exposure  to a plateau
concentration  of  =0.5  ng/m2.  after  6  hours.   A  similar  pattern  of uptake was
observed  with other lipophilic  solvents (i.e.,  carbon  tetrachloriue,  hexane,
tetrachloroethylene-, 1,1,1-trichloroethane, and trichloroethylene).
     Absorption of toluene vapor through the  skin  does  not appear to result in a
significant contribution to the body burden of toluene as  compared to absorption
through the respiratory tract.  In experiments conducted by Riihimaki and Pfaffli
(1978), volunteers wearing light, loose-fitting clothing and respiratory protec-
                                                                 "oT." i
tion were exposed  to  600  ppra  toluene for 3-5 hours.   The subjecJS^emained at
rest except for 3 exercise perio-ds, each lasting  for  10 minutes, which occurred
at 0.5, 1.5,  and 2.5 hours of exposure.   The exercise was sufficient  to stimulate
perspiration  and  raise the  skin  temperature  slightly,  conditions  which  are
thought  to  enhance percutaneous absorption.   The concentration  of toluene in
peripheral venous  blood, measured at  the end of  1, 2,  and  3  hours  of exposure,
Was constant  at approximately 100 (ig/i.
     Riihimaki  and  Pfaffli  (1978)  compared total  uptake  through  the  skin
(calculated from  the  amount of  toluene  exhaled  assuming that  16$ of absorbed
toluene  is  exhaled)  with  theoretical  uptake through the  respiratory  tract
(assuming pulmonary ventilation  of  10  Jl/min  and  retention  of 60?)  at the same
(600 ppm) level  of exposure.   They estimated that  uptake through  the skin was
approximately  1$ of the theoretical uptake through the  respiratory  system.
     In  similar  experiments conducted by Piotrowski  (1967, reviewed in NIOSH,
1973)i subjects exposed dermally to  1600 mg/m-' Ci27  ppm) toluene for 8 hours had
no  increase  in urinary excretion of  a metabolite  (benzoic  acid)  of toluene.
Based  on  this result,  Piotrowski  (1967) concluded that  absorption of toluene
                                      13-8

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through the skin would  not exceed 5% of absorption through the respiratory tract
under the saine conditions.
     The absorption of toluene from the gastrointestinal tract appears to occur
more slowly than through the respiratory tract,  but appears to be fairly complete
based on experiments with  animals.   The concentration  of  radioactivity in the
blood of adult male rats reached a maximum  2 hours  after gastric intubation of
100- \jJi   4-%-toluene in  400 u& peanut oil  (Pyykko  et al.,  1977).   The oil may
have  retarded absorption.    Based  on  the  percentages  of  the dose  excreted
unchanged  in  the  expired air  and as  hippuric acid  in the urine  of rabbits,
toluene  appears to  be completely  absorbed  from  the   gastrointestinal  tract
(El Masri et al.,  1956; Smith et al.,  195*4).
13.2.  DISTRIBUTION
     Toluene  is highly soluble  in lipid  and sparingly soluble in  water,  as
indicated  by  the  partition  coefficients  in  Table 13-2.   Judging from  the
fluid/air  partition  coefficients for  water,   plasma,  and  blood,  much  of  the
toluene in  blood may be  associated  with the lipid  and  lipoprotein  components,
including  the cellular elements.   The  tissue/blood partition  coefficients for
fatty tissues  were very high (113 for adipose tissue  and  35 for bone marrow); for
other tissues, they ranged from about  1 to 3.
     Little is known  about the  tissue distribution of toluene in humans.  During
inhalation exposure to 50 to 200 ppm toluene,  the slow approach to steady-state
of  peripheral  venous  concentrations   as  compared  to  arterial  concentrations
(described  under absorption) indicates  that equilibration with the  tissues may
take at least 2 to 3 hours.  Concentrations in peripheral  venous blood do not,
however,  reflect  the  dii. harge  of toluene to the  tissues  as fully  as would
concentrations in central venous blood.   A teenage boy  who  died from sniffing
glue had the following levels of toluene in his tissues:  heart  blood, 11 mg/kg;
liver, 47 mg/kg; brain, 44 mg/kg;  and  kidney,  39 mg/kg  (Winek et al. 1968; also
reported in Winek and Collum,  1971).
     Several laboratories have investigated the tissue  distribution of toluene
and its metabolites in animals exposed by inhalation to  relatively high concen-
trations of toluene.  The concentrations  of  toluene in liver, brain,  and  blood of
mice exposed  to 3950 ppm  (15 mg/fc) toluene for  3 hours in  a  dynamic exposure
Chamber rose continuously throughout the exposure period, as  shown previously in
Figure 12-1.  Concentrations of toluene reached 625  mg/kg in  liver,  420 mg/kg in
brain, and  200  mg/kg  in blood  at  the end  of  exposure  (Peterson and Bruckner,
                                      13-9

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                                   fABLE 13-2

                   Partition Coefficients for Toluene at 37 °C
                                 Partition  Coefficient   Reference
I.  Fluid/Air or Material/Air

    Water
    Oil, olive
    Blood, Human
    Fat, human, peritoneal

    Oil, olive
    Lard
    Blood, human

    Blood, human
    Blood, rabbit
    Plasma, rabbit

II.  Tissuea/Blood (Rabbit)

    Liver
    Kidney
    Brain
    Lung
    Heart
    Muscle, femoral
    Bone narow, red
    Fat, retroperitoneal
   2.23
 492
  15.6
1296

1380
1270
  15.6

  K.64
  10.41
  16.99
   2.58
   1.54
   3.06
   1.92
   2.10
   1.18
  35.43
 113.16
Sato and Nakajima, 1979a
Sherwood, 1976
Sato et al., 1974a, 1974b
Sato et al., 19?4a, 1974b
 Homogenatfcs.

 20 J  fat by volume.

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1978;  Bruckner and Peterson, 198la).  Exposure of mice  to 10,600 ppm (40 mg/£)
toluene for   10 minutes  resulted  in  lower  tissue  and  blood  concentrations.
Intermittent  exposure  to  10,600  ppm in cycles of 5 minutes on, 10 minutes off or
10 minutes on, 20 minutes off for  a total  of  3 hours produced tissue and blood
levels approximately 3 times-higher than those produced  by the single 10-minute
exposure to 10,600 ppm and similar  to those produced by the 3-hour exposure to
T.0,600 ppm. The intermittent exposures were an attempt to simulate solvent abuse
(e.g.,  glue  sniffing) by  humans  (Peterson  and  Bruckner,  1978;  Bruckner  and
Peterson,  198lb).
     From an  analysis of the time  course  of  toluene levels  in  central venous
blood and  in brain of rats during and after whole-body exposure of the aniials to
575 ppm toluene,  Benignus  et al.  (1981) concluded  that their data  fit a one-
compartment model.    Groups  of rats  were  killed  at   intervals  from   15  to
240 minutes  during  exposure  and  at  intervals  from 15  to  2^0 minutes  after
exposure.  Blood samples were taken from the  posterior  vena cava at sacrifice.
According  to  the one-compartment  model analysis  of Benignus  et  al.  (1981),
asymptotes  (steady  state  levels)  during  exposure  to  575  ppm   toluene  were
10.5 ppm  toluene  in blood and  18  ppm  toluene in brain,  and concentrations of
toluene in these tissues reached 95? of  these  estimated asymptotes in approxi-
mately  55 minutes.  Visual  inspection of the  experimental data,  however, shows
that the mean toluene concentrations in the blood and brain of rats exposed for
120 and 2^0 minutes  were above the  predicted asymptotes  and, particularly i;i the
brain,  may still  have been increasing appreciably  during' this  interval.   The
authors mention that this observation "could be construed as  an indication that a
multicompartment model ought to  have been fitted."  The elimination of toluene,
estimated by one-compartment model analysis, occurred at a slightly faster rate
from the brain than from central  venous blood.  The experimental data of Eenignus
et al.  (1981) for rats are similar to those of Peterson and Bruckner (1978) for
mice, previously discussed.
     After adult male rats  were exposed  by inhalation to radioactively-labeled
toluene, the  highest  concentrations of radioactivity were found in their white
adippse tissue  (Carlsson and Lindquist,  1977;  Pyykko  et al.,   1977).   In the
experiments of Pyykko and  coworkers (1977) the  concentration of  radioactivity
reached  a maximum  in  all  tissues,  but  white   adipose tissue  within  15 to
30 minutes after the end of 10 minute?  of exposure to  ^600  ppm  ll- H-toluene.  The
concentration in white adipose tissue reached a maximum  1 hour after the  end of
                                     13-

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exposure.    In the  experiments of  Carlsson  and  Lindquist  (1977)F  a  similar
increase in the concentration of radioactivity in white adipose tissue occurred
during  the first  hour  after  cessation  of  exposure  for   1 hour  to  550 ppra
(1.950 mg/O methyl-  C- toluene.   No such increase occurred in other tissues.
     Carlsson and Lindquist  (1977)  found  that after white  adipose  tissue,  the
next highest concentrations of  radioactivity  occurred  in adrenals and kidneys,
followed by  liver,  cerebrum,  and cerebellum.   At the  end  of  exposure,  white
adipose tissue contained a  6-fcld higher concentration of radioactivity than did
cerebrum or cerebellum.  Pyykko et  al. (1977) reported that after white adipose
tissue, the next highest  concentration  of  radioactivity  was  found in  brown
adipose tissue,  followed  in  order  of  decreasing  concentrations by  adrenal,
stomach, liver and kidney,  brain and other tissues, blood, and bone marrow.  The
loss of radioactivity from  adipose tissue  and  bone marrow appeared to occur more
slowly than the loss from other tissues  (Pyykko et al.,  1977).  Radioactivity in
the tissues presumably represented  toluene and its metabolites.
     Bergman (1979),  using three-stop whole-body autoradiography, investigated
the  distribution  of toluene,  its  metabolites,   and  covalently  bound  reactive
                                                                           ill
intermediates in mice exposed to an extremely high concentration of methyl-  C-
toluene.  This work was briefly described in a previous report (Bergman, 1978).
The mice were exposed singly to a very high initial concentration of toluene for
 10 minutes in a closed chamber,  as described in Section  13.1., and sacrificed at
intervals  thereafter.    Low temperature  autoradiography,  performed at  -80°C,
allowed the detection of both volatile radioactivity (representing toluene) and
non-volatile  radioactivity  (representing metabolites).    In  a second  step,
sections   were   dried   and  heated   to   remove   volatile  material   before
autoradiography, thus permitting detection of  non-volatile metabolites only.  In
the third step, sections that had been dried  and  heated were then extracted to
 remove  water-soluble  and lipid-soluble i-adioactivity,  presumably leaving on] y
 the radioactivity that was  covalently bound to proteins and nucleic acids.
     Lov;  temperature  autoradiography  performed  immediately  after  exposure
revealed high levels of radioactivity in adipose  tissue,  bone marrow, and soinal
 nerves, with some radioactivity also present  in  the  brain,  spinal cord, .liver,
 and kidney  (Bergman, 1979).   Bergman  reported that the adrenal  did not contain
 high  concentrations  of radioactivity,  but  he did  not  discuss  whether radio-
 activity was found in the stomach.

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    The  only  radioactivity visible in  dried,  heated sections appeared  in  the
liver,  kidney, and  blood  (Bergman,  1979).   This  indicates that  significant
amounts of metabolites  had  already  been  formed  by  the end of exposure,  and that
the radioactivity in fat  and nervous tissue was  due  to the parent  compound,
Similarly,  as  early  as  8  minutes  after  intraperitoneal  injection of  290 \ig
  C-toluene/kg into mice, the majority of  radioactivity in the kidney  (78$)  and
liver  (64J)  and  about  half  the  radioactivity  in  blood C<8J) was  reported to
represent non-volatile metabolites, while most of the radioactivity in brain and
virtually all  in the adipose tissue  was volatile and  thus  represented toluene
itself  (Koga,  1978).  The methods used in Koga's study  are  unclear  because  the
t?xt of  the  paper is  in Japanese, with only the figures, tables,  and summary in
English.   Bergman (1979)  reported  that  no radioactivity was  detected  in auto-
radiograms  prepared from dried,  heated,  and extracted  sections,  indicating an
absence of covalent binding.
     As nad  been  observed in the studies of Pyykko et al. (1977) and Carlsson and
Lindquist (1977), radioactivity disappeared from the tissues relatively quickly
after  exposure was terminated.  The  distribution patterns observed in mice killed
more than 4  hours after exposure were the same on low temperature  autoradiograms
as on  dried, heated sections.   Thus,  the radioactivity remaining  in the tissues
at this  time represented  non-volatile metabolites.   At  8 hours after  exposure,
only  the kidney  and  the  intestinal   contents  had detectable  radioactivity
(Lergmam, 1979).
     Oral a-dainistration  of iJ-  H-t.oluene (100 \il toluene in  400 uE, peanut oil by
intubation)  to aault male rats  produced a pattern of tissue distribution similar
to  that  produced  by  inhalation exposure  (Pyykko  et al.,  1977).   Distribution
appeared  to  be delayed,  however, by  absorption from the digestive tract.  Maximum
tissue  concentrations  occurred  2   to  3 hours  after  administration   for  scat
tissues  and  5  hours after administration for adipose tissue.
     In  sunmary,  toluene was preferentially accumulated  in adipose tissue  and
was retained longer in adipose tissue  and  bone marrow than  in  other tissues,
which  is reasonable   on   the  basis  of  the  high  tissue/blood  distribution
coefficients of  these  tissues.   Toluene  and   its  metabolites  were   found in
relatively high concentrations  in tissues active in  Its metabolism and excretion
(i.e., liver and kidney).   Levels in the  brain relative  to those in other tissues
Were perhaps  lower  than woull be expected on the  basis  of  the tissue/blood
                                     13-13

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distribution  coefficients  reported  by Sato  et  al.  (197^a,  197^b).   Tissue
distribution was  similar after inhalation and oral exposure.
13.3.   METABOLISM
    Toluene  is  thought to  be metabolized  in  humans  and  in  animals  by  the
pathways  outlined in Figure  13-1.    Some  of the  absorbed toluene  is excreted
unchanged in the  exhaled air,  but the major portion is metabolized by side-chain
oxidation to benzole acid, which is  conjugated with glycine to form hippuric acid
and then  excreted in  the urine. Small  amounts  of  benzoic acid may be conjugated
with glucuronic  acid.   Minor  amounts  of  toluene undergo  ring hydroxylation,
probably  via are.rie oxide intermediates, to  form o-cresol and p-cresol, which are
excreted  in the urine as sulfate or glucuronide conjugates.
     Hurans exposed  to toluene  by  inhalation exhaled  about  16? of the absorbed
toluene  after  exposure  was  terminated,  according  to Nomiyama arid  Nomiyaaa
(197^) and Srt-ova and  Teisinger (1952,  1953), - or  4J, according to Veulemans and
Masscheleir. (1978a:.  Volunteers inhaling 50 to  150 ppm toluene for about t hours
during rest or exercise excreted 60 to  70%  of the  absorbed dose as hippuric acid
in the urine during  and after exposure  (Veulemans  and  Masschelein,  1979).   A
siarilar va^ue  was obtained when subjects were  exposed  to toluene (67 ppm) and
xyleris (85 ppa)  simultaneously for  3  hours;  68J of  the  absorbed  toluene  was
excretec  as urinary hirDuric acid during and after exposure (Ogata et  al., 1970).
Srbava and Teisin.ger  (19l~3)  reported that  although most of  the benzoic acid in
tre urine of subjects who inhaled 72  to 532 ppm (0.271  to 2.009 mg/fc)  toluene was
excretec.as nippuric  acid,  10  to 205 was  excreted as a glucuronide conjugate.
     The  excretion of hippuric acid in  the urine was elevated within 30 minutes
of  the initiation of  inhalation  exposure,  indicating  that the  metabolism of
Uluci.-e is rapid  (Noraiyama and  Norciyama,  1975;  Ogata et al.,  1070; Veulemans and
Masschelein,  1979).   The maximum rate  of hippuric acid formation from benzoic
acid was  reporter by Amsel  and Levy  (1969) to be about  190  [jmol/nin,  and it
appeared  to be limited  by  the availability  of glycine  (Amsel  and  Levy,  1969;
Quick.  1931).   Assuming retention of  60? of the  inhaled concentration, Riihimaki
(1979)  estimated  that uptake of toluene may saturate the conjugation  capacity at
a  toluene  concentration of  760 ppm  (32  mmol/m-1) during  light  work  (pulmonary
ventilation of 10 fc/roin) or 270 ppm  (11  mmol/m-1) during  heavy  work  (pulmonary
ventilation of 30 Z/min).
     o-Cresol,  a  cocpound that  is  not  detected often  in normal urine, has been
identified in  the urine  of workers exposed  to 7 to  112  ppm  toluene  (Angerer,

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 EXHALED
UNCHANGED
                      CH-,
                    o
                  TOLUENE
                                      CHjOH
    o   -
BENZYL ALCOHOL
                                   i-CRESOL
                                                                  CONMCHjCOOM
                                                                6
                                                             HIPPURIC ACID
                                                              GLYCINE
                                                        COOH
                                                    BENZOlC AGIO
                                                            GLUCURONIC ACID
                                                    BEN20YL GLUCURONIDE
                                                    GLUCURONIDE AND
                                                    SULFATE CONJUGATES
        Figure 13-1.   Metabolism of  Toluene In Humans  *nd Animals
                       (Adapted from  Lahaia, 1970)
                                    13-15

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1979;  Pfaffli  et al.,  1979;  Hansen,  1982).   The concentration  of  _o-cresol  in
urine  collected  at  the end of exposure  was  directly proportional to  the time-
weighted  average  exposure  of  the workers  (Pfaffli et al., 1979).  Angerer (1979)
estimated that approximately  0.05% of the retained  toluene  had been  metabolized
to ci-cresol.  £-Cresol also may have been a metabolite of  toluene, as its concen-
tration was  higher  in  the  urine  of workers exposed  to toluene than in the urine
of  unexposed  workers  (Angerer,  1979).   The  difference,   however,  was  not
significant.   Wiowode reported  finding  jn-cresoi in  addition to _o-  cresol  and
£-crescl  in the  urine of  workers  exposed to 280 ppm  toluene  (Woiwode  et  al.,
1979.)  and male  subjects who were experimentally exposed  to  200 ppm  toluene for
4 hours (Woiwode and Drysch,  1981).  No  ni- cresol  was  detected in the urine of
unexposed workers or the  subjects  before the experimental  exposure.   No other
studies of ^r,  vivo  human or animal metabolism or _in vitro microsooal metabolism
reviewed  for this document have  detected _m-cresoj.  as a metabolite of toluene.
     The  concentration of  pnenol has been reported to be slightly elevated in the
urine  of  exposed workers  as  compared to controls  (Angerer,  1979;  Szadkowski
et al., 1973/-   The origin of the  increased  phenol  excretion was thought to be
the sTsall  amount of  benzene present  in industrially-used  toluene  (Angerer,
1^79).
     The  metabolism of toluene  has  been more fully  studied in animals than in
h'jma.-is. T*"'e initial .itep in the metabolism of toluene to  benzoic acid appears to
be sice-chajn hydroxyLation of toluene to benzyl  alcohol by the microsorual mixed-
function  oxidase system.  Toluene has been  shown to  produce a type  I binding
spectrum   v«ith cytochrome  P45Q from  rat? and hamsters,  indicating that  it  is
probably  a substrate for the mixed-function oxidase system (Canady et al., 1974;
Ai-Goilany eL al.,  1978).  When incubated with rabbit  hepatic microsomes, toluene
was metabolized primarily  to benzyl alcohol (Daly et al.,  1968) and small amounts
of benzyl alcohol have been detected  in  the  urine  of rats  given toluene orally
(Bakke and Sneline,  1970).
     Additional  evidence that toluene is metabolized by mixed-function oxidases
has been obtained by Jkeda  and Ohtsuji  (1971) who demonstrated  that the induction
of  hepatic mixed-function oxidases  by  pretreatment of  adult female  rats  for
1 days witn phenobarbital increased  the metabolism  of toluene.   When  given
1. IB ing  toluene/kg  body   weight   intraperitoneally,  phenobarbital-pretreated
(induced) rats had  greatly  elevated  urinary excretions  of  hippuric  acid and
decreased concentrations   of  toluene  in  the  blood compared  to non-induced rats
                                     13-16

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given the same dose of toluene.  Induced i^ats had high levels of benzoic acid in
the blood;  non-induced rats had none (blood was obtained at decapitation).
     The increased metabolism of toluene by induced rats appeared to reflect an
increase in side-chain hydroxylation of  toluene, because the activity of hepatic
sides-chain   hydroxylase,  assayed,  in vitro  with  the model  substrate  p-nitro
       i                              ~~~~~~~
toluene, was significantly increased per gram of liver.   The i_n vitro oxidation
of the  resultant  alcohol  (p-nitrobenzyl alconol)  to the  acid  vp-nitrobenzoic
acid) was not affected. The conjugation of benzoic acid with glycine, measured
in vivo as the total  amount of hippuric acid excreted after  benzoic acid adminis-
tration, was also unaffected (Ikeda and Ohtsuji, 1971).
     It has been assumed (Ikeda and Ohtsuji,  1971;  Nomiyama and Nomiyama, 1978;
NRC, 1980), by analogy with the metabolism of the model substrate p-nitrotoluene
(Gillette,  1959), that benzyl alcohol  is metabolized to benzaldehyde by alcohol
dehydrogenase and  that benzaldehyde  in turn  is  oxidized to benzoic  acid by
aldehyde dehydrogenase.  These  enzymes  both are.found  in  the  soluble fraction
from liver.  Benzaldehyde  itself  has not  been  detected  in the urine or expired
air  of  animals  given toluene  orally  (Smith et al.,  1951*; EUkke  and Sheiine,
1970).   Metabolism  of  toluene probably occurs primarily  in the  liver.   This
assumption is based on the  previously  discussed tissue  distribution of metabo-
lites, the demonscrated metabolism of  toluene by liver microsomal preparations,
and by analogy with  the metabolism of  other xenobiotics.
     Rabbits intubated  with 300 mg toluene/kg  body  weight eliminated approxi-
mately 18$ of the dose in  the expired air  (Smith et  al.,  195*0  and, in another
study from the same  laboratory, excreted about 7*t? of the  dose as hippuric acid
in  the  urine (El Masri  et  al.,  1956).  These results  are  similar  to those
obtained with humans  who  inhaled toluene.   None of  the toluene appeared to be
converted to benzoyl glucuronide (Smith et al., 195*0, although about T4J of an
oral dose of benzoic acid  was  excreted  by  rabbits  as the  glucuronide conjugate
(Bray et al., 1951).
     Toluene  metabolism  appears   to  be  rapid in animals,  as  shown  by  the
appearance   of metabolites in  the livers,  kidneys,  and  blood  of  mice within
minutes of   exposure  to  toluene  (Bergman,  1979;  Koga,  1978)   (discussed in
Section 13.2.) and by the increase.'  Tinary excretion of  hippuric acid in rabbits
within 0.5  hour  of the initiation  of inhalation  exposure  (Nomiyama  and Noraiyama,
1978).  As  was   previously mentioned  for  humans,  the  rate of  conjugation of
benzoic acid  with glycine  may be  limited  in  animals  by the  availability of
                                     13-17

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glycine.  Administration of glycine to dogs exposed by inhalation to 200, 100, or
600 ppm toluene enhanced  the  rate 01  hippuric  acid excretion  (Von  Oettingen,
19H2b). At tne end of  8 hours of exposure to 600 ppm toluene, the concentrations
of toluene in peripheral  venous blood from glycine-treated dogs were lower than
the concentrations in dogs  that had  not  been treated  with glycine.   No  such
difference was observed at the  two lower exposure levels.  This result suggests
that  conjugation  of  benzoic  acid with  glycine may  have limited  metabolic
elimination at the highest  level  of exposure.  The level  of  exposure at which
glycine treatment produced  a difference in venous  blood levels of  toluene is
similar to that (780 ppm) calculated  by Riihimaki (1979) for  saturation of the
glycine conjugation capacity of humans.
     A minor  pathway  for the  metabolism of  toluene  is ring  hydroxylation by
mixed-function oxidases.   Incubation of toluene with rat or rabbit liver micro-
somes resulted in the  production of small amounts of ^-cresol and _p_-cresol (Daly
et al., 1968; Kaubisch et al.,  1972).  The migration  of deuterium  when toluene
was labeled in the ^-position and  a comparison of the  rearrangement products of
arene oxides of toluene with the  cresols obtained by microsomal metabolism of
toluene indicated  that  arere  oxides  are  intermediates in the metabolism  of
toluene to o- and £-cresols  (Daly  et al.,  1968;  Kaubisch et al., 1972).
     Because phenols,  including cresols, are eliminated in the urine as sulfate
conjugates, thereby increasing  the excretion of organic sulfates and decreasing
the  excretion  of  inorganic  sulfate,  investigators have used  urinary sulfate
excretion  after toluene administration as an indicator  of cresol formation.  Oral
doses of 350 mg toluene/kg  body weight  produced  no  increase in organic sulfate
excretion   in  rabbits  (Smith et al.,  195*1).    In rats,  high  doses  (2.2  and

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macromolecules.   Very little toluene is metabolized  via  this  pathway,  however,
and the studies  already discussed  in  the  distribution  section  indicate  that
binding of  toluene  metabolites  to proteins and nucleic acids  does  not  occur to
any significant  extent.
     Van Doorn and  coworkers (I960) have  reported  detecting small amounts  of a
mercapturic acid, tentatively identified  as  benzylmercapturic  acid (N-acetyl-S-
benzyl-L-cysteine),  in the urine of  male rats treated with toluene.   Approxi-
mately O.i; to  0.7$  of a dose  of 370 mg/kg  toluene  body weight,  administered
intraperitoneally,  was recovered as the mercapturic acid.  The concentration of
glutathione in the liver was decreased slightly by  administration of  toluene.
Benzylmercapturic acid  would  arise  from  conjugation with  glutathione of an
electrophilic  product of side-chain oxidation of toluene.
     The metabolism  of  toluene appears  to  result  in its  detoxification.   The
length of  the  sleeping time produced by high doses of toluene (1.18 to 1.45 g/kg
intraperitoneally)  was decreased in phenobarbital-induced female rats to 50? or
less of the sleeping time of  controls  (Ikeda  and Ohtsuji,  1971).  Similar results
were  obtained with  male mice  (Koga  and  Ohmiya, 1978).   Phenobarbital-induced
animals, however,  did not  have  significantly different mortality rates  than
controls when given  high doses  of  toluene  (Ikeda  and Ohtsuji,  1971;  Koga and
Ohmiya, 1978). Male  mice given various inhibitors of drug metabolism (SKF 525A ,
cyanamide,  and pyrazole)  30 minutes before the injection  of toluene had sleeping
times that were significantly longer than those  of control  mice and had higher
mortality  rates than did control mice (Koga and Ohmiya,   1978).
13.1.  EXCRETION
     In both  humans  and  animals, toluene is  rapidly excreted as the unchanged
compound in expired 'air  and  as  a metabolite, hippuric acid, in the urine.  Most
of the absorbed toluene  is  excreted within 12 hours  of the end of exposure.
     The concentrations  of toluene  in  exhaled air  and   in arterial  and venous
blood of human subjects  declined very rapidly as  soon as  inhalation exposure was
terminated (Astrand et al.,   1972;  Carlsson  and Lindquist,  1977;  Ovrum et  al.,
 1978; Sato et  al.,  1974b; Veulemans and Masschelein,  1978a,  1973b).  Sato et al.
(197^b) reported that  semilogarithmic   plots   of   toluene  concentrations  in
alveolar air and  in  peripheral  venous blood versus time after the end of exposure
suggested  that desaturation  occurred in  three exponential phases:   an initial
rapid phase, followed by an  intermediate  phase,  and then  a slow phase.  The  data
were obtained  from  3  male subjects who  inhaled 100 pptn toluene  for 2 hours  (Sato
                                     13-19

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et al.,  1974b; clarified in Sato and Nakajima,  1979b).   The desaturation curves
were  resolved  graphically  into three components, and constants  were determined
by the least squares method.   The rate  coefficients  and  corresponding half-lives
(t.,?) for the decay of toluene in  peripheral venous b? ood were 0.355 min   ^1/2
= 1.95,.'minutes),  0,0197  min~1  (t..,- =  35.2 minutes), and 0,00339 min"   (t1/2
204 minutes).   Rate  coefficients  and  half-lives  for  the  decay of  toluene  in
alveolar air  were   0.437 min"1   (t,/2   -   1-5^ minutes),   0,0262  min"   (t-i/o
  -26.5 minutes),  and  0.00313 min'1  (t1/2    221  minutes).
     Because the  rate coefficient  for  the rapid phase was  derived  from only two
points  (at-C and  5 minuces), the second of  which belonged  with the  intermediate
phase,  Sato et  al.  (1974b)  noted  that  the  coefficient   for  the   rapid  phase
invol%'ed some  error.   The  data of Sato et al. (1974b) indicate that the decay of
toluene  concentrations in  peripheral venous blood was more gradual  than that in
expired  air.   Similar conclusions have been reported by Astrand et al. (1972),
and Veulemans  and Masschelein  (1978b).  Astrand et  al. (1972) have  reported that
peripheral  venous  concentrations  declined more  gradually  than   did  arterial
concentrations .
     Veulemans end Masschelein (1978a) and Nomiyama  and Nomiyama  (1974b)  found
the excretion  curves for toluene in expired  air  to be adequately described as the
smr,  of  2 exponential terms  rather than  3-    Subjects  for these  studies  were
exposed  to ^0, 100, or 1^0 pptn toluene for about 4  hours.  The sampling regimens
differed from that  of Sato  et  al.  (1974b),  in that Veulemans  and Masschelein
 (19?&a)  did  not  begin monitoring expired air as soon after exposure ended, and
Nomiyama and Noitiyama (1974b)  sampled expired air infrequently during the period
used  by  Sato et al.   (1974b) to determine  the first two exponential  phases.  Rate
coefficients   for the rapid  and  slow  phases  were  calculated by Veulemans and
Maaschelein  (19783)  to be 0.340 min   and  0.00608 min   ,  respectively,  using a
curve-fitting  computer program.  These rate coefficients  corresponded to half-
lives of 2.04 and  114 minutes.   Nomiyama and  Nomiyama (1974b) reported  rate
coefficients  for  the rapid phase of 5.10 h~   (**    - 8.16 minutes) for men and
      _ 1
3-22  h   (t    - 12.9 minutes) for women;  the rate coefficient for ti e slow phase
was 0.335 h~1  U     - 124  minutes)  for both sexes.
     In  the  desaturation period, men  and women expired 17.6  and  9-4?, respec-
tively,  of  the total amount  of toluene' calculated  to have been absorbed during
exposure (Nomiyama and Nomiyama, 1974b).  These  values are close to  what had been
reported previously  (i.e., 16J) by Srbova and Teisinger  (1952,  1953) in abstracts
                                     13-20

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from the foreign literature.  .Veulemans and Masschelein  (1978a)  estimated that
about 4$ of the  toluene absorbed during exposure was subsequently excreted in the
expired air.  Unlike  the  continuous  exposures employed in the  other  pertinent
investigations,   however,   the  exposure  regimen  employed  by  Veulemans  and
Masschelein (19?8a) was  discontinuous (i.e.,  four 50-minute periods of exposure
separated by 10-minute intervals of nonexposure).
     According to Veulemans and Masschelein (1978a)  a  much  greater variability
was observed for the excretion of toluene in expired air during the  first 4 hours
after the end of exposure than had been observed for the related lung clearances
during exposure. This variability could be explained  partially by differences in
respiratory  minute volume  during  the  post-exposure  period;  the percent  of
absorbed toluene  iroreted in the  expired air  during  the first 4 hours  after
                                                                     2
exposure  correlated  positively  with  respiratory minute  volume  (r   -  0.7D.
Another  factor  that appeared to affect  excretion was  the amount  of  body fat,
because  there was  a  significant (p  < 0.025)  negative  correlation  between  fat
content  as measured by the  index of  Broca and the percent excretion in expired
  I                             2
air  after  exposure at rest (r   =  0.2134).   This indicates that  less of  the
absorbed toluene would be excreted  in the  expired air of an obese person than in
the  expired  air of a thin person during the  first  4 hours  of  desaturation.
Additionally, subjects who had been exposed to toluene while exercising expired
less of  the absorbed  amount during the  first  4 hours  of  desaturation than  did
subjects who had been exposed while resting (Veulemans  and Masschelein, 1978a).
     As previously described,  60 to 70? of the toluene absorbed by humans  during
inhalation can  be  accounted for as  hippuric  acid in  the  urine  (Veulemans  and
Masschelein, 1979;  Ogata et al., 1970).  The excretion rate of hippuric acid in
the urine of subjects inhaling 50,  100,  or 150 ppm toluene increased during the
first 2 hours, leveling  off at about  the third hour after initiation of exposure
(Veulemans and Masschelein, 1979; Nomiyama and  Nomiyama,  'i978).   Hippuric acid
excretion  (mg/hr)  declined  fairly  rapidly after cessation of about k hours of
exposure.     Nomiyama and  Nomiyama  (1978),  treating  this   decline   as  a
monoexponential process, determined  a  half-life for hippuric acid in urine of
117 minutes for men and  74 minutes  for women.   Veulemans and Masschelein (1979)
reported an  initial,  fairly rapid decrease  with a  half-life  between 2.0  and
2.3 hours,  followed  by  a more  gradual  return to baseline  excretion  levels by
about 24 hours after the start of exposure.
                                     13-21

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     The excretion rate  of  hippuric acid, measured at the end of about 4 hours of
experimental exposure or 8  hours of occupational exposure, correlated reasonably
well with the  uptake rates  (Veulemans  and Masschelein,  1979) or  total  uptake
(Wilczok and Bieniek,   1978) during  exposure.   At  a  given level  of  physical
activity and exposure concentration,  the intra- and interindividual variability
in hippuric  acid excretion was  greater than that noted for uptake rates and was
attributed to the variable  baseline excretion of this compound  because it was not
explained by other factors  (body weight, body fat, cardiorespiratory parameters)
.(Veulemans and  Masschelein, 1979).  Exercise during exposure increased the rate
of excretion of  hippuric•acid  (Veulemans and Masschelein,  1979) in  acccrdance
with the increase in uptake rate.
     Hippuric acid is a  normal constituent of urine derived from benzoic acid and
precursors of benzoic acid in the diet (Quick,  1931).  Concentrations of hippuric
acid in  thfe urine of 101 workers not  exposed to toluene  ranged from  0.052 to
1.271 mg/mJl  (corrected to urine specific gravity of 1.02^)  and rates of excretion
of 'hippuric  acid  ranged from 18.47 to  23-00 mg/hr for diuresis  of  greater than
30 mSYhr (Wilczok and Bieniek,  1978).   Others .have also  reported  great  varia-
bility in the physiological concentrations  of  urinary hippuric  acid  (Ikeda and
Ohtsuji, 1969;  Ir.amura  and  Ikeda,  1973;  Engstrom, 1976;  Kira,  1977;  Ogata and
Suglhara, 1977;  Angerer, 1979).
     Volunteers exposed  in a chamber  to 200 ppm toluene for 3 hours followed by a
1 hour  break and an additional  4 hours of  exposure excreted hippuric acid as
shown in Figure  13-2 (Ogata et  al.,   1970).  This exposure regimen was chosen to
simulate exposure in the  workplace.  After  leveling off  after  approximately
3 hours of  exposure, excretion  increased again  during the afternoon exposure.
The rate of hippuric acid  excretion  remained elevated for  about 2 hours  after
exposure was terminated  and then declined almost to baseline levels by 18 hours
after the end of exposure.  The total quantity of hippuric acid  excreted during
the  period  lasting  26  hours  from   the  initiation  of  exposure  was  directly
proportional to  the degree of exposure  (ppm x time) up through the  highest
toluene concentration of 200 ppm and could be used to calculate  exposure with a
fairly high  degree of accuracy.  Less accurate  for  this  purpose were excretion
rates  during exposure   (i.e., total  hippuric acid  excreted during  exposure +
 time)   and  concentrations  in   urine,    corrected   for  specific   gravity.
Concentrations  of hippuric  acid in  urine collected during the  entire exposure
period and  corrected to a specific   gravity of  1.021)  were 0.30  + 0.10,  2.55 +
                                     13-22

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                                                               -« CONCENTRATION (mg/mi)
u>
I
ro
u>
                     ~ 12-,
                      z
                      o
                      cc
                      I-
                      z
                      a)  c j
                      o  6-
                      z
Q  4-|

o


u

E  2H

D
                                                          .	o  RATE (mg/minute)
                                           6    8    10   12   14   16   18   20  22   24   26


                                                         HOURS
                                                                             Z _

                                                                             2  g
     U H
     < D



-4   E I





-2   £
                                                                                            i  L0
                       Figure  13-2.  Urinary Concentrations and Excretion  Fates of Hippuric Acid  in

                                    Volunteers Exposed  to Toluene (Volunteers were exposed to 196 ppm

                                    toluene for 3 hours in the morning and  for A hours in the after-

                                    noon with one hour's break in between.   Points are means + SEH.)

                                    (Ogata et aj.. ,  1970)

-------
0.55,  and 5.99 ±  1.20nig/niS,  (mean + standard deviation) for  control,  100 ppro,
and 200  ppm  exposed subjects,  respectively.  Values for controls were lower and
more uniform than  those reported by others, as described previously^
     Spot urine samples  collected  from workers after at least 3 hours of exposure
to toluene (and from nonexposed workers at the sarqe time) have not given as good
a distinction between unexposed and exposed  workers.   Imamura and Ikeda (1973)
have pointed out tnat the upper fiducial limit (P ^  0.10) of' normal hippuric acid
concentrations, whether or  not  corrected for  specific gravity, is so close to the
lower, fiducial limit of workers exposed to 100 ppm toluene (the Threshold Limit
Value)  that  such  a   measurement  would   not  be  reliable   in  screening  for
overexposure.   This conclusion was based on  data  reported  jy Ikeda and Ohtsuji
(1969).   The correlations between  concentrations of toluene in workplace air and
the•concentration  of  hippuric acid in  urine of  individual  workers  have  been
       i
relatively poor (Veulemans  et  al., 1979;  Szadkowski,  1973;  Ogata et al.,  1971).
The  correlation   between  exposure  concentration  and  excretion  rate  during
                                                            o
exposure, although  slightly   better,  was   also  poor:    r1"  -  '">.096  for  the
correlation  with hippuric  acid  concentration (corrected for  specific  gravity)
and  r^    0.116 for  the correlation  with rate of excretion  of hippuric  acid
(Veulemans et  al.,  1979).  Some of the variance in excretion rates was accounted
for by differences in lung clearance, and,  hence, uptake among  workers (Veulemans
et  al.,  1979).
                                                                Ill
     Mice exposed  to a  very high initial concentration of methyl-  C-toiuene in a
closed chaaber for  10 ciinutes excreted about  10? of the  absorbed dose as volatile
material in  the exnaled air and about  68J  as  unidentified compounds in the urine
within  8 hours   (Bergman,  '.979).    Details of   exposure were  discussed  in
Section 13.1.   In  these experiments, volatile expired radioactivity (thought to
represent the  parent compound) was collected continuously  in  a trapping device.
The total volatile radioactivity expired during each time interval was converted
to the roear,  percent dose excreted  per minute  during that interval and plotted at
the end cf the interval. The resultant semilogarithmic plot of mean percent dose
exhaled per  minute versuj time was a curve.  Computerized  non-linear regression
analysis  of  the  data  according   to  the  method  of   least  squares  yielded  3
exponential   components  with   rate  coefficients  of  0,0659,   0.0236,   and
0.0014 min~    corresponding   to  apparent   half-lives  of   10.5,   29.^,   and
 158.7 minutes, respectively.

-------
     The  respiratory  rates  of  the  mice  were,  according  to  Bergman  (1979)f
"remarkably  reduced"  during exposure,  and.  nence probab.ly were reduced during at
least part, of  the  post-exposure period.  If respiratory minute volumes were also
.decreased,  thJLs. would,  on  the  basis  of  the  observations of  Veulemans  and
Masschelein  (1978a),  be  expected to reduce the  pulmonary  excretion  of toluene.
TheWsults of Bergman (1979) nay  therefore not  be relevant to exposures at lower
concentrations  of  toluene.
     After inhalation exposure of rats  or  mice to toluene, the  disappearance of
toluene and  its metabolites from blood and from most  tissues,  including brain,
was  rapid (Peterson  and Bruckner,  1978;  Benignus  et  al.,   1981; Carlsson  and
Lindquist, Pyykkc  et al., 1977; Bergman,  1979)  as described in Section 13.2.  The
exceptions  were  white   adipo.se  tissue,  for  whicn   both  accumulation  and
elimination  were  slow,  a_'id  bone rr.irrow,  for which  elimination  was  very slow
(Carlsson and  Lindquist,  19''?; Pyykko et al .,  1977).  By 2U hours after exposure
to radioactively-labeled toluene, the concentration  of  radioactivity remaining
in most tissues was less  than 1.  and that  regaining in adipose tissue was about
5J of the initial  whole-body concentration (Pyykko et al., 1977).
     Rabbits exposed  to  toluene- vapor at 55U ppm for  100 minutes or 4500 ppm for
10 minutes had increased rate? of urinary hippuric  acid excretion that reached
maximum  values   l.h hours   after*   exposure   (Nomiyana   and  Nomiyaaa,  1978).
Excretion rates returned to  baseline levels at  7  hours  after the initiation of
exposure  to  350 ppzi for  100  minutes  and at about 3 hours after the initiation of
exposure  to  4500 ppm  for  10 tLinutes.
     Dermal  exposure  of human subjects to toluene liquid or vapor resulted in the
appearance of  toluene in the expired air  (Guilleraan et.al.,  197**;  Riihircaki and
Pfaffli,  1978) as  discussed  in Section- 13.1.  The  excretion of  toluene in the
expired air of subjects exposed  to 600 ppm  toluene for  3 hours appeared  to
consist of at least 2 exponential phases (ffiihimaki and Pfaffli,  1978).  The mean
amount of toluene  expired during the "quantitatively significant" portion of the
excretion curve was calculated to be 45.9 umole (4.23 mg) Riihimaki and Pfaffli,
1978). Piotrowski (1967, reviewed in NIOSH,  1973)  found  that  subjects exposed
dennally   (with respiratory  protection)  to  427 ppm  (1600  nig/or) toluene  for
8 hours  had  no   detectable  increase  in   urinary  excretion of  benzoic  acid
(presumably  analyzed  after  hydrolysis of  conjugates).
     Oral administration of toluen^ to rabbits resulted in a pattern of excretion
similar to thai observed after inhalation exposure  of  humans.   Rabbits (N = 2)
                                     13-25

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intqbated  with 350 mg  toluene/kg  body weight  expired  18J of  the dose  as
parent  compound within  1^.5 hours; less than  ^% of the dose was  eliminated in the
expired air in the period from 1^.5 through 35 hours after dosing (Smith et al.,
1951)).   In similar  experiments from  the same laboratory,  rabbits intubated with
274 mg  toluene/kg body weight excreted an  average of  7'4f of the  dose in the urine
as  hippuric  acid;  excretion  was  complete  with 2^ hours  of  dosing  (El  Masri
et al., 1956).  The  elimination of toluene and its metabolites from tissues and
blood  of  rats  given toluene  orally  (Pyykko et  al.,  1977)  was similar  to  the
pattern already described after inhalation exposure  (Pyykko et  al., 1977) except
that elimination after oral  administration  appeared to  be delayed  by a slower
rate of absorption  than had  been observed for inhalation exposure.
   '.The  excretion  of  other  metabolites   of  toluene  (i.e.,   cresols,  benzyl
alcohol,  glucuronide  and  sulfate  conjugates,   benzylmercapturic  acid) in  the
urine of humans and animals has already been described in  Section 13-3-  With the
possible exception  of benzoylglucuronide (Srbova and Teisinger,  1953),  none of
these  excreted metabolites  represented more  than  about  1?  of the  total dose of
toluene administered or absorbed  (Ar.gerer,  1979; Bakke  and  Sheline,  1970;  Van
Doom  et  al.,  1980;  Smith  et al.,   195^).    Trace  amounts  of  toluene  were
eliminated in the  urine  of  humans  exposed   to  toluene   (Srbova and  Teisinger,
1952).
     Biliary  excretion of  toluene  or  its  metabolites appeared to be negiigiole.
                ill
Rats given 50 mg   C-toluene/kg body weight intraperitoneally excreted less than
2%   of   the   administered    radioactivity   in   the   bile    within   2*4 hours
(Abou-El-Markarem et al.,  1967).
     Most of  the  experimental work on  the disposition  of toluene  in humans  and
animals has  focused on single exposures.  The elimination of  toluene  is  rapid
enough   that  few  investigators  have  studied  its  potential  accumulation  with
repeated daily exposure.   Ovrum and coworkers  (1978)  took  samples of capillary
blood daily  before  work from  8 printers exposed occupationally to 35 to 353 ppm
toluene.  No  cumulative increase  in  blood concentrations of toluene  was found
during  the course of a  5 day work week. Konietzko and ooworkers  O9&0) observed,
however,  that  toluene concentrations in  peripheral  venous  blood  tended  to
increase during the  course of a 5  day work week, although the ranges overlapped
(Table  13-3). Mean  exposure  concentrations,  measured by  a personal air sampling
method, did  not  increase during the week.   The blood samples were taken before
work on Monday, Wednesday,  and Friday from 8 workers exposed to 18I< to 332 pptn
                                     13-26

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                                                    TABLE  13-3
             Toluene Concentrations in Workplace Air and Peripheral  Venous Blood of Exposed Workers
a,b


First week
r
ro
-j
Second week


Toluene in air (ppm)
Toluene in blood
before exposure (jig/ml)
Toluene in blood
after exposure (ng/mi)
Toluene in air (ppm)
Toluene in blood
before exposure (^g/mi.)
Toluene in blood
after exposure (ug/nul)
Monday Tuesday
225 233
(95-303) (153-383)
0.12
(0.09-0.21*)
3.63
(2. 3-1*. 75)
285 304
(145-473) (190-521)
0.27
(0.07-0.57)
11.60
(6.99-17.10)
Wednesday
209
(107-341)
0.51
(0.28-0.82)
6.69
(4.21-10.36)
309
(213-413)
1.00
(0.35-151)
10.49
(3. 24-20. 3D
Thursday Friday
212 203
(92-314) (124-309)
0.77
(0.29-1.
6.70
(3.99-10
232 191
(125-451) (105-432)
1.21
(0.44-2.
5.85
(1-94-9.


67)
.6?)
29)
73)
Source:  Konietzko et al., 1980
Means and (range) of eight workers

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daily in a plastic processing  factory.   Concentrations in  blood  samples  taken
after work were  highly variable arid did not seem to follow a consistent  pattern.
     In an analysis of  3155 samples of urine taken  in  the  course  cf biological
monitoring from  different workers  on  different  days  of  the week and in different
workplaces,  Lenhert et  al. (1978)  observed that concentrations of hippuric acid
in\ the  urine  did  not  vary with the day of  the week except  on Monday,  when the
concentrations  were  significantly  higher than  on  other  days.   Tha  authors
conjectured that the elevation of  hippuric acid concentrations on Mondays  was a
result of different: eating habits  on the weekend.
     In  experiments  with dogs, exposure to  400  ppm for 7 hours  a day  for 5
consecutive days did net result in an increase in the  total  amount of  hippuric
acid  excreted per  day  over the period  of 5 days or change the time course of
urinary excretion (Von Cettingen et al.,  19^2b).   Nor  did  the concentration of
toluene in peripheral venous blood sampled at the end of exposure increase with
dayiof exposure.
13.5.  SUMMARY
     Toluene is readily  absorbed  through the respiratory tracts  of  humans and
expennental  animals,   as  would   be  expected  from its  blood/air  partition
coefficient of approximately  15 (Sato and Nakajima, 1979a;  Sato et al.,  197^3,
197^b; Sherwood, 1976).   The amount of toluene absorbed (uptake) is proportional
to  the  concentration  in inspired  air,  length  of.  exposure,  and  pulmonary
ventilation (respiratory  minute volume)  (Astrand et al., 1972; Astrand,  1975;
Veulemans and Masschelein, 1978a).
     The  uptake  of  toluene by humans  was  about 50? of  the  amount  inspired
(Veulemans and Masschelein, 1978a; Carlsson  and  Lindqui.3t,  1977.  Ovrun et al.,
1978).   Total   uptake    (absorption)    can   be    approximated   as   follows:
Uptake = 0.5 V  C.  t, where V   is  the respiratory minute volume in JL/min,  C. : s
the inspired concentration  in mg/S,,  and t is the length of exposure in minutes
(Ovrum  et  dl.,  1978;   Veulemans   and  Masschelein,  1978a).     Because  of  its
dependence on respiratory minute volu-me,  the  uptake of toluene is affected by the
subjects' l*>vel  of  physical  activity  (Astrand  et  al.,  1972; Astrand,  1975;
Veulemans and Masschelein,  1978a;  Carlsson and Lindquist,  1977).   A  subjects'
content of adipose  tissue  had little or no effect  on  the uptake of toluene during
exposures lasting M hours or less  (Veulemans and Masschelein, 1978a; Astrand et
*!•»   1972)  except in the case of  extremely  obese individuals  (Carlsson and
Lindquist,  1977),  zvnd  even then  the increased  uptake  may have  been  at  least
                                     13-28

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partly due  to greater pulmonary Ventilation  in  the obese subjects than  in  the
thin ones.  Under  "steady state" conditions,  peripheral venous concentrations of
toluene correlated roughly  with exposure  concentrations.    Inter- and  intra-
individual  variability were  high enough to make  this an insensitive estimate of
exposure  concentration or uptake  (Von Oettingen  et al., 19*42a,  19*»2b;  Veuleaans
and Masschelein,  197db).
     Although toluene appears  to  be absorbed less readily through the  skin than
through the respiratory tract,  percutaneous absorption  of liquid toluene may be
significant.  The tnaxitausi toluene  concentration in  peripheral  venous  blood of
subjects  who imaersed one hand in liquid toluene for 30  minutes was about 26$ of
the  concentration in peripheral  venous blood of subjects who  inhaled  TOO  pptn
toluene vapor for 30  minutes (Sato and Naxajima,  1978).  Absorption of  toluene
vapor through the skin in humans,  however,  probably anounts  to less  than 5? of
the  total  uptake  through the respiratory  tract under the  same-  conditions  of
exposure (Riihimaki  and  Pfaffli,  1976;  Piotrowski,  1967;  reviewed  in  NIOSH,
1973)'  Absorption of toluene through  the  gastrointestinal tract  appears  to be
fairly complete,  based on the amounts of toluene and  its metabolites excreted by
experimental animals  after  administration  of  toluene  (Pyykko  et al..  1977;
El Hasri  et al.,  1956; Smith et  al.,  1951).
     Toluene appears  to  be  distributed  in the  body  in  accordance  with  the
tissue/blood distribution coefficients and  its  metabolic  and  excretory  fats.
Thus, toluene itself  is found in high concentrations in adipose tissue and bcne
marrow, and toluene and its  metabolites are found in moderately high  concentra-
tions in liver and kidney (Peterson  and Bruckner,  1978;  Bruckner and  Peterson,
198la; Carlsson  and Lindquist,  1977;  Pyykko  et  al.,  1977; Bergoan,  1979).   The
time course of toluene concentrations in  the brain  appeared  to correlate with
behavioral  effects (Petersor and Bruckner,  1976; Bruckner and Peterson,  198la).
     The major portion of inhaled  or  ingested  toluene is metabolized by side-
chain oxidation  to benzoic acid, conjugated with glycine  to  form hippuric acid,
and excreted in  the urine.   Regardless of  the route of  administration, dose, or
species,  60 to 75J of the absorbed (inhalation)  or administered (oral)  toluene
could be  accounted for as  hippuric  acid in  the urine  (Veulemans and Masschelein,
1979; Ogata et al., 1970; F.I Masri et al.,  1956).  Much  of the remaining toluene
(9 to  18J) was  exhaled  unchanged  (Nomiyama and  Noraiyama,  19?1b; Srbova  and
Tsisinger,  1952,  1953; Smith et al.,  195M.  Two percent or less appeared in the
urine as  cresols  and benzylmercapturic acid; these  metabolites  are of  concern
                                     13-29

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because they indicate formation of reactive intermediates that potentially could
bind to  tissue macromolecules.    No evidence  of  covalent  binding  to tissue
components  has  been detected, however, by autoradiography  of mice that inhaled
iaC-toluer.e (Bergman,  1979).
     Most of  the toluene absorbed by humans  or animals after inhalation or oral
exposure  is excreted within 12 hours  of the end of exposure (Ogata et al., 1970;
Veuleaans and Masschelein, 1979; Nomiyama and Nomiyama, 1978;  Smith et al., 1951;
Bergman,  1979).  In experimental animals, elimination  of toluene and its metabo-
lites from most tissues,  including brain,  was rapid;  elimination from fat and
bone qaarrow was  slower (Peterson  and  Bruckner,   1978;  Benignus  et  al.,  1981;
Bruckner  and  Peterson,  198la;  Pyykko  et  al.,   1977;  Carlsson  and Lindquist,
1977).
     In humarLS , tr.e time  course  of desaturation  after  cessation of inhalation
exposure  appeared to consist of  3 exponential phases  with  half-lives  of 1.95,
35.2, and 20^ minutes  for toluene concentrations  in peripheral venous blood and
1.59, 26.5, and  221 minutes  for  toluene concentrations in  alveolar  air (Sato
et ol.,  i9'ii).   Toluene concentrations in expired  air  or peripheral venous blood
after the  encJ of  inhalation  exposure were  not  reliable indicators  of toluene
•uptake  or  of  exposure  concentrations  because of the great  variability among
individuals (Veulemans  and Masschelein,  1978a,  1978b;  Astrand et  al., 1972).
Sone of *.:u s variability, particularly in expired air concentrations,  could be
explained ty Differences in exercise  loaa during exposure, in respiratory minute
volumes a: ter exposure, and in adipose tissue content  (Veuleraans and Masschelein
'978a,  i97b:,;.   Siitilariy, although the excretion of hippuric acid ir; the urine
is roughly proportional to the  degree of  exposure to toluene, inter- and intra-
ini.Viduai  variations  in  tr.e physiological  excretion of hippuric  acid render
quantitatior.  of exposure or  uptake  n'rom urinary  hippuric acid concentration or
excretion rat<=c unreliable (InsEazura and  Iksda,   1973; Vetileaians  et al., 1979;
Veulemans ar.d Masschelein, 1979;  Ogata et  al., 1971; Wilczok and  Bienick, 1978;
arid otners  as reported in Section I^.U.).

13.6  REFERENCES

ABOU-EL-KAhKAREM,  M.rt. et al.  (1967).  Biliary excretion of foreign compounds.
Benzene and its  derivatives  in  the rat.   Biochem. J.   105:1269-1274.
                                     13-30

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AL-GAILANY,  K.A.S.,  HOUSTON,  J.B.,  and  BRIDGES,  J.W.   (1978),    The role  of
substrate  lipophilicity in determining Type  1 microsomal PM50 binding  charac-
teristics.   Biochem.   Pharmacol.   27X50:783-788.

•AMSEL, \L.P., and LEVY,  G.   (1969).  Drug  biotransTormation interactions  in man.
II.  A  pharaacokinetic study of  the  simultaneous  conjugation  of benzoic  and
salicylic  acids with  glycine.  J.  Pharm.  Sci .   58(J) :321-326.

ANGERER, J.   (1979).   Occupational chronic exposure to organic solvents.   VII.
Metabolism of  toluene in man.  Int.  Arch. Occup.  Environ.  Health.   ^3( 1 ):63-67.

ANGERER,  J. and  BEHLING,  K.   (1981).   Chronische Loosungsmittelbelastung  atn
Arbeitsplatz:   IX. Ein Verfahren  zur Evalvierung von  Grenzwerten  fur  parameter
der in'neren Belastung am  Beispiel  der  Toluolexposition.   Int.  Arch. Occup.
Environ. Health.   _|48:  137-1^6.   (Cited in Apostoli  et  al., 1982).

APOSTOLI,  P.,  BRUGNONE, F.,  PERBELLINI,  L., COCHEO,  V.,  BELLOMO,  M.L.  and
SILVESTRI,  R.   (1982).  Biomonitoring of Occupational toluene exposure.   Int.
Arch.  Occup. Environ.   Health.   50:  153-168.

ASTRAND,. I.    (1975).   Uptake  of  solvents  in  the  blood and tissues of man.  A
review.  Scand. J_. Work Environ. Health.   KM: 199-218.

ASTRAND,  I.,  EHRNER-SAMUEL,  H. ,   KILBOM,  A.,  and OVRUM,  P.   (1972).   Toluene
exposure.    I.   Concentration in  alveolar  air and  blood ?.t rest and during
exercise.  Work Environ. Health.   72(3):119-130.

BAKKE, O.M., and SCHELINE, R.R.  (1970).   Hydroxylation of aromatic hydrocarbons
in the rat.  Toxlcol.  Appl.  Pharmacol. J6:691-700.

BENIGNUS,  V.A., MULLER, K.,  BARTON,  C.N.  and  BITTIKOFER,  J.A.   (1981).  Toluene
levels  in  blood  and  brain  of rats  during  and  after   respiratory  exposure.
Toxicol. Appl.  Pharmacol.   6±: 326-334.
                                     13-31

-------
BERGMAN,  K.   (1978).   Application of whole-body autoradiography to distribution
studies of organic solvents.  Int. Symp. Control Air Pollut* Work. Environ. Pt.
2, pp. 128-139.

BERGMAN,  K.   (1979).  Whole-body  autoradiography anc allied tracer techniques in
distribution and elimination studies of some, organic  solvents.   Scand.  J_.  Work
Environ.  Health.  _5:263 pp.

BRAY, H.G.,  THORPE, W.V., and WHITE, K.   (1951). Kinetic studies of the metabo-
lism of foreign organic compounds.  Biochea. Jl.  *48: 68-96.

ERUCKKiR, J.V., and PETERSON, fi.G.  (1978).  Effect  of repeated exposure of mice
and raus to concentrated toluene and acetone vapors.  Toxicol . Appl .  Pharmacol .
^5(1): 559.

BhuCKKES, J.V., and PETERSON, R.G.  (198la).  Evaluation of toluene and acetone
inhalant,  abuse.   I.    pharmacology  and  pharmacodynamics.    Toxieol .  Appl .
Fnaraacol .  t_i : 302-312.

BFUCKNEh, J.V., and PETERSON, R.G.  (198lb).  Evaluation of toluene and acetone
inhalant abuse. II.  Model development and toxicology.  Toxiccl Appl.  Pnarmacol .
6jl: 27-38.

CANAL:,  W.J.,  ROBINSON, D.A.,  and COLBY,  H.D.   (1974).   Partition  model for
hepatic  cytcchronie p-ii^O-hydrocarbon complex  formation.    Biocheta.  Pharmacol .
 CARL3SON, I'., and LINDGUIST,  T.   (1977).  Exposure of animals and man to toluene.
 Scand.  J. Wo-k, Environ. Health.  3( 3) : 135-1 ^3.

 CAHL££ON, A. and LIHDQUIST, E.  (19&2).  Exposure to toluene.  Concentration in
 subcutaneous adipose  tissue.   Scand ^. Work,  Environ. Health.   6(1): 56-62.
 Taken from: Chem. Abst.  96:  212050y,  1982.
                                     13-32

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CARLSSON, A.  (1982),  Exposure to toluene.  Uptake,  distribution  and  eliminatioh
in man.  Scand. J. Work, Environ. Health.  8(1):  ^3-55.  Taken from:  Chem. Abst.
96: 2120196,  1982.

CHEMICAL INDUSTRY INSTITUTE OF TOXICOLOGY  (CUT);   (1980).  A  twenty-four month
inhalation  toxicology study in Fischer-S^M  rats  exposed  to atmospheric toluene.
Executive Summary and Data Tables.  October 15,  1980.
       ,  T.,  EDFOR'S, M.-L.,  and RYLANDER, R.   (1968a).   Mouth  absorption of
various  compound  sin cigarette  smoke.  A_rsh. Environ.  Health.   16(6) ;831-635.

DALHAMN,  T.,  EDFORS, M.L.,  and RYLANDER,  R.   (1968b).  Retention of  cigarette
smoke components  in  human lungs.   Arch.  Environ. Health.   17:7^6-7^6.

DALY, J., JERINA, D.,  and  WITrCOP B.   (1968).  Migration  of deuterium during
hydroxylation of  aromatic substrates by liver microsomes.  I.  Influence of  ring
substituents.   Arch. Biochem.  Biophys.   128( 2) :517-527 .

DUTKIEWICZ, T., and  TYRAS,  H.   (iy68a).   The quantitative estimation of  toluene
skin absorption in man.  Arch.  Gewerbepath Gewerbehyg.  24:253-257-

DUTKIEWICZ, T., and  TYRAS, H.  (1°68b).  Skin absorption of toluene, styrene, and
xylene by man.  Brit. J. Ind. Med.  25(3):2^3.

EGLE, J.L. and GOCHBERG, B.J.   (1976).  Respiratory retention of inhaled toluene
and benzene in the dog.  _J.  Toxicol . Environ.  Health.   1 ( 3):531-538.

EL MASRI,  A.M., SMITH, J.N., and WILLIAMS, R.T.  (1956).  Studies  in  detoxifica-
tion. The metabolism of alkyl benzenes, _n- pro py] benzene , and n-butyl benzene  with
further  observations on  ethylbenzene.  Biochem. J_.   64:50-56.

ENGSTROK,  K.,  HUSMAN, K., and RANTAKEN,  J.  (1976).  Measurement  of  toluene and
xylene metabolites  by gas  chromatography.   Int.  Arch. Occup. Environ.  Health.
36(3) : 153-1 60 /
                                     13-33

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GERARDE, H.W., andAHLSTROM, D.B<  (1966).  Toxicplogic studies  on  hydrocarbons.
XI.  Influence of dose on the metabolism of mono-n-alkyl derivatives of benzene.
Toxic.  Appl. Pharm.  2:185-190.

GILETTE,,. J.R.   (1959).   Side chain oxidation of p^nitrotoluene;   I.   Enzymatic
oxidation of  p-nitrotoluene.  ^J. Bio'l. Chem.  23*1:139-1
GUILLEMIN, M., MURSET,. J.C., LOB, M., and RIQUEZ, J.   (1974).   Simple  method to
determine the efficiency  of  a  cream used for skin protection against  solvents.
Brit. J_. Ind.  Med.  31 (4):310-316.

HANSEN, S.H.  and  DOE3SING, M.   0982).  Determination of  urinary hippuric acid
and o-cresol, as indices  of  toluene  exposure,  by liquid  chroiratography  on
dynamically modified silica.  £. Chromatgr.   229:  141-148.

IKEDA,  M.,  and OHTSUJI,  H.    (1967).    Significance of  urinary  hippuric  acid
determination as  an index of toluene exposure.   Brit.  J_.  Ind. Med.   26:244-246.

IKEDA,  M.,  ar.d OHTSUJI,  H.    (1969).    Significance of  urinary  hippuric  acid
determination  as  an   index  of   toluene   exposure.     Brit.   J.  Ind.   Med.
IKEDA,  M.,  and OHTSUJI, H.   (1971).   Phenobarbital-induced  protection  against
toxicity  of  toluene  and   benzene in  the  rat.    Toxicol.   Appl.   Pharmacol.
20(1).-30-43.

IMAMURA,  T.,  and  IKEDA,  M.  (1973).  Lower fiducal  limit  of  urinary  metabolite
level as  an index of excessive exposure to industrial  chemicals.  Brit,  jj.  Ind.
Med.  _30:269-292.

JAKOBSON, I., WAHLBERG, J.E., HOLMBERG, B. and JOHANSSON, G.   (1982).  Uptake via
the blood and elimination of 10 organic solvents following epicutaneous exposure
of anesthetized guinea  pigs.  Toxicol. Appl.  Pharmac.^1.   63:  181-187.
                                     13-34

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KAUBISCH,  N., DALY, J.W., arid JERINA , D.M.   (1972).   Arene oxides  as  interme-
diates in  the  oxidative metabolism  of  aromatic  compounds.   Isomerizatipn  of
methyl-substituted arene oxides.   Biochem.  1 1 : 3080-3088.

KIRA, 3.   (1977).  Measurement by gas chromatography of urinary hippuric acid and
methylhippuric  acid as indices of  toluene and  xylene exposure.   Brit. J_.  Ind .
Med.  BM1*): 3C5-309.  Taken from:   Chem. Abst.   88:841370,  1978.
KOGA, K.  (1978).  Distribution,  metabolism and excretion of toluene  in  mice.
Folia Pharmacol. Jpn.  7*t( 6) : 687-698 .

KOGA, K., and OHMIYA, Y.   (1978).   Potentiation of toluene toxicity by hepatic
enzyme inhibition in mice.   J_. Toxicol. Sci.  3( 1 ) :25-29.

KONIETZKO, H.,  KEILBACH,  J.,  and DRYSCH,  K.    ('1980).   Cumnmlative  effects  of
daily toluene exposure.   Int.  Arch.  Occup. Environ. Health.  *l6( 1 ) :53-58.
LAHAM, S.  (1970).  Metabolism cf industrial solvents.  Ind.  Med.  _39:  61-6*1.

LEHNERT, G., R.D. LADENDORF arid  D.  S2ADKOWSKI.   (1978).   The relevance  of  the
accunulation of  organic solvents  for  the  organization  of  screening tests  in
occupational medicine.  Int. Arch. Occup-Environ. Health.  |4_1:  95-102.

NIOSK (NATIONAL INSTITUTE FOR  OCCUPATIONAL SAFETY AND HEALTH).  (1973). Or. teria
for a Recommended Standard.   Occupational Exposure to Toluene.   Final  Report.
Contract No. HSM-99-72-118.  Available through NTIS No. PB-222-219/8,  108 p.

NOMIYAMA, K., and NOMIYAMA, H.  (1978).  Three fatal cases of thinner-sniffing,
and experimental  exposure  to  toluene in human and  animals.   Int.  Arch.  Occup.
Environ.  Health.  tj1( 1): 55-6*1.

NRC (NATIONAL  RESEARCH  COUNCIL).   (1980).   The Alkyl Benzenes.   Committee  on
Alkyl Benzene Derivatives,  Board  on  Toxicology and Environmental Health Hazards;
Assembly of Life Sciences,  National  Research Council.  Washington, DC:  National
Academy Press.
                                     13-35

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\
 NOMIYAMA*  K., and NOMIYAMAji  H.   (197^a).   Respiratory  retention,  uptake and
 excretion  of organic solvents  in  man.   Benzene; toluene, n-hexane, trichloro-
 ethylene,  acetone,  ethyl  acetate  and  ethyl  alcohol.   Int.  Arch. Arbeitsmed.
 32(1-2):75-83.

 NOMIYAMA,  K., and NOMIYAMA,  H.   (197Mb);   Respiratory elimination of  organic
 solvents in man.   Benzene,  toluene,  n-hexane,  trichloroethylene,  acetone,  ethyl
  kcetate and ethyl  alcohol.   Int. Arch.  Arbeitsmed.   32(1-2):85-91.

 OGATA,  M., TAKATSUKA, Y.,  TOMOKUNI,  K.,  and MUROI,  I.   (1971).   Excretion  of
  hippuric acid and  m- or  p-methylhippuric acid in the  urine of persons exposed  to
  vapors  of  toluene  and m- or  p-xylene in  an exposure chamber and in workshops,
  with  specific  reference   to  repeated   exposures.     Brit.   J^.  Ind.  Med.
  23(4):382-385.

  OGATA,  M.,  TOMOKUNI,  K.,  and TAKATSUKA,  Y.    (1970).    Urinary excretion  of
  hippuric acid and  m- or  p-methylhippuric acid in the urine of  persons exposed  to
  vapors  of  toluene  and m- or p-xylene as a test of  exposure.   Brit. ^J.  Ind. Med.
  27(1):43-50.

  OGATA,  M., and SUGIHARA, R.  (1977).  An improved direct colorimetric method for
  the quantitative analysis of urinary hippuric  acid as an index  of toluene  expo-
  sure.  Acta.  Med. Okayama.  31:235-242.
  OVRUM, P.,  KULTENGREN,  M.,  and LINDQUIST,  T.  (1978).   Exposure to  toluene  in  a
  photogravure printing  plant.   Concentration in ambient  air  and uptake in  the
  body.  Scand.  J_.  Work,  Environ. Health.  H( 3): 2 37-2*45.

  PETERSON, R.G., and BRUCKNER,  J.V.   (1978).  Measurement of toluene levels in
  animals tissues,   In:   Voluntary Inhalations of Industrial Solvents.  C.W. Sharp
  and Carrol,  L.T., editors.   Rockville, MD:  Nat.  Inst.  Drug Abuse.   2£:33-^2.

  PFAFFLI, P.,  SAVOLAINEN,  H.,  KALLIOMAKI,  P.L.,  and KALLIOKOSKI,  P.   (1979).
  Urinary  c-cresol  in  toluene   exposure.    Scand.  ^J.  Work   Environ. Health.
  5(3):286-289.
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FJOTROWSKI,  J.  (1967).  Quantitative  estimate  of the absorption of toluene in
people.  Med. PraOy.  _^8:213-223-  (In Pol.) (Cited in NIOSH, 1973).

PYYKKO, K., TAHTI,  H.,  and VAPAATALO, H.   (1977).   Toluene  concentrations ifi
various tissues of rats  after inhalation and oral administration.  Arch. Toxicol.
3J5:169-176.   Taksn from:  Chem. Abst. 88:'45927r,  1978.

QUICK, A.J.   (193D-   The conjugation of  bepzoic acid in the urine.  J_. Biol.
Chem.  92:65-85.

RIIHIMAKI, V.,  and PFAFFLI,  P.    (1978).   Percutaneous  absorption  of solvent
vapors in man.  Scand. ^J. Work Environ. Health.   4( 1 ): 73-85.

RIIHIMAKI, V.  (1979).  Conjugation and urinary excretion of toluene and m-xylene
metabolites in a man.  Scand. ^J. Work Environ. Health.  5(2): 135-142.

SATO,  A.,  FUKIWARA,  Y.,  and  NAKAJIMA,  T.   (197^b).  Solubility  of  benzene,
toluene and tD-xylene in  various body fluids and tissues of  rabbits.  Jap. J_, ind .
Health.  _16(1):30.

SATO,  A., and NAKAJIMA, T.  (1979a).   Partition coefficients of some aromatic
hydrocarbons  and  ketons-s   in  water,  blood  and  oil.    Brit.  .J.  I_nd.  Hed .
SATO,  A.,  and  NAKAJIMA,  T.   (1-?79b).   Dose- dependent  metabolic  interaction
between benzene  and  toluene iri vivo  and  in vitro.   Toxicol .  Appl .  Pharmacol .
SATO,   A.,   NAKAJIMA,   T.,   FUJIWARA,   Y.,   and   HIROSAWA,   K.     (197ta).
Pharmacokinetics of benzene and toluene.   Int. Arch.  Arbeitsmed.  33( 3): 169-182.

SATO, A.  and  T. NAKAJIMA.   (1978).   Differences  following skin or inhalation
exposure  in  the absorption  and  excretion kinetics  of  trichloroethylene  and
toluene.  Brit. J.   Ind. Med.  35: 1)3-49.
                                     13-37

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SHERWOOD,  R.J.   (1976).   Ostwald solubility coefficients of  some  industrially
important  substances.   Brit.  ^J.  Ind.  Med.   33(2): 106-107.

SMITH,  J.N. et al.   (1954).   Studies  in  detoxication,  55.   l^e metabolism  of
alkylbenzenes.   a/Glucuronic acid  excretion following  the administration  of
alkylbenzenes; b/Elimination  of  toluene in  the expired air of rabbits.  Biochem.
J.  56:317-320.

SREOVA,  J., and TEISl'NGER,  J. (1952).   Absorption and elimination of toluene  in
nan.  Arch. I_n£.  H yg.  Occup.  Med.  6^:462.

3RECVA,  J., and  TEISINGER,  J.   (1953). Metabolism of toluene.   Pracov.  Lek.
5_:259-263.  Taken from:   Chem. Abst.  ^9:34186,  1955.

SZADKOWSKI, D.,  PETT,  R. , ANGERER,  J., MANZ,  A.,  and  LEHNERT,  G.   (1973).
Chronic solvent  exposure at  work.   II.  Harmful material  levels in  blood and
excretion  rates  of  metabolites  in urine with  the   importance of  environmental
criteria for toluene exposed  printers.   Int.  Arch.  Arbeitsmed.   31(4):265-276.

VAN DCORN, R., BOS,  R.P.,  and BROUNS,  R.M.E.   (1980).   Effect  of  toluene and
xyleneo on liver  glutathione  and  their  urinary excretion as mercapturic acids  in
the rat.  Arch. Toxicol.   43( *Q :29?-304.

VERSCHUEREN,  K.    (1977).  Handbook of  Environmental Data  or. Organic Chemicals.
New York,  NY:  Van Nostrarid Reinhold Company, pp. 592-596.

VEULEKANS, H., and  MASSCHELEIN,  R.   (1978a).   Experimental human  exposure  to
toluene.  I.   Factors influencing the individual respiratory uptake  and elimina-
tion.  Int. Arch.   Occup. Environ. Health.   42(2):105-117.   Taken  from:   Chem.
Abst.  _90:l8l.040q, 1979-

VEULEMANS, H. and MASSCHELEIN,   R.   (1978b).   Experimental human  exposure  to
toluene.  II.   Toluene in venous  blood during  and  after exposure.   Int.  Arch.
O^ccup.  Environ.   Health.   42(2):  105- i7. Taken from:  Chem. Abst.  90: I8l040q,
1979.
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VEULEMANS,  H., and  MASSCHELEIN,  R.   (1979).   Experimental  human exposure  to
toluene.   III.   Urinary  hippuric  acid excretion  as a  measure  of  individual
solvent  uptake.   Int.   Arch.  Occup.  Environ.  Health.   43(1):53-62.

VON OETTINGEN, W.F., NEAL, P.A. .and DONAHUE, D.D.   O?'i2a).   The toxicity and
potential  dangers of toluene—Preliminary report.  £. Am. Hed. Assoc.   118: 579-
584. ^.

VON OETTINGEN, W.r.,  NEAL;  P.A.;  DONAHUE,  D.D,,  SVIRBELY,  J.L., BAERNSTEIN,
H.D., MONACO,  A.R.,  VALAER, -P.O., and MITCHELL,  J.L.  (19^2b). The Toxicity and
Potential  Dangers of Toluene, with  Special  Reference  to  its Maximal Permissible
Concentration.  U.S. Public Health  Serv.  Pub. Health  Bull. No. 279, 50  pp.

WILCZOKv T.,  and BIENIEK,  G.  (1978).  Urinary hippuric acid  concentration after
occupational  exposure to toluene.   Brit. £.  Ind. Med.  35(1):330-33H.
     i
WINEK, C.L.,  and COLLOM,  W.D.   (1971).   Benzene  and  toluene fatalities.    J_.
Occup.  Med.   J^:259-261/

WINEK, C.L.,  WECHT,  C.H.,  and COLLOM,  W.D.   (1968).  Toluene  fatality from glue
sniffing.   Perm. Med.  71:81.

WOIWODE, W.,  WODARZ, R., DRYSCH, K., and WEICHARDT,  H.   (1979).   Metabolism  of
toluene in man:  Gas  chroraatographic  determination of o~.  m-. and p-cresol  in
urine.  Arch. Toxicol.  j43:93-98.

WOIWODE, W.,  and DRYSCH, K.   (1981).  Experimental  exposure to toluene.  Further
consideration of cresol formation in man.   Br.  J. Ind. Med.   38:  19^-197.
                                     13-39

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            T4.  CARCINOGENICm, MUTAGENICITY, AND TERATOGENICITY
11). 1.  CARCINOGENICITY
    CUT  (1930) concluded  that exposure  to  toluene at levels  of  30,  100, or
300  pppn  for  6  hours/day,  5 days/week for 24 months did not produce an increased
incidence  cf neoplastic,  proliferative, inflammatory or degenerative lesions in
Fischer  3^  male or female rats  (see description  of  study in Section 12.1.2.).
Neoplasms  were  observed  frequently in  the lungs  and  liver,  endocrine organs,
lymphoreticular system, mammary  gland,  integument,  testis and  uterus,  but the
lesions  occurred with equal frequency in all control and treatment groups.
    Although  the CUT (1980)  study was comprehensive and is  the only chronic
bioassay of  toluene presently  available,  it  should  be  noted  that  there are
several  factors  that  preclude  a definite conclusion  of non-carcinogenicity.
First,  it should  be noted  that  this study has been  considered inadequate for
carcinogenic!ty evaluation (Powers,  1979)  because  a maximum tolerated dose  (MTD)
was not  achieved at  the highest  dose tested in either this  2-year study (300 ppm)
or in a  preliminary  90-day study (1000  ppta).  Also, the low mortality of rats in
the CUT  study (1*4.6%)  differs from  the  mortality  rate (up  to 25?)  normally
associated with maintaining these animals under barrier conditions (NCI, 1979a,
b).  If  the CUT animals  were not raised under barrier conditions (which is not
stated), then  still higher mortality rates would be expected in  this age group of
Fischer  314*4 rats.    A  high  spontaneous  testicular  interstitial  cell   tumor
incidence in aging F3t*14  rats  (66 and 85J reported by  Coleman  et al.,  1977 and
Mason  et  al.,  1971, respectively)  removes  this  organ frotc any assessment of
carcinogenicity.  Additionally,  the high spontaneous  incidence  (165)  of  mono-
nuclear  ceil  leukemia  on aging F3^ rats  (Coleman  et al.,  1977) suggests that
this strain may be  inappropriate for the study  of  a chemical that may be  myelo-
toxic  (Table  12-7).  An  independent quality  assurance audit of the CUT  study
indicated that there were no deviations from protocol  requirements, and that the
final  report  accurately   reflects   (with  minor exceptions)  the data  from the
laboratory records.  The errors that were noted in  the final report, do not affect
the conclusions drawn frotn the data.
     A  chronic bioassay of commercial-grade toluene in rats and  mice exposed by
inhalation  is currently  being  conducted  by  the  NTP Carcinogenesis  Testing
Program.  Prechronic testing of commercial  toluene in the same species exposed by
gavage  has been completed (NTP,  1983).
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    Toluene has been utilized extensively as a solvent for lipophilic chemicals
being  tested for  their carcinogenic  potential  when  applied topically  to the
shaved  skin  of  animals.  Results of control  experiments  with pure toluene have
been uniformly  negative.   Peel  (1963) r  f°r example,  applied toluene (volume not
stated)  to the  shaved  Interscapular skin 3 times a week throughout the lifetime
of 54 male SWR, C3HeB, and A/He mice and found no carcinogenic response.  Coombs
et al.  (1973) treated the dorsal skin of 20 randomly bred  albino mice with 1 drop
of toluene (6 nt)  twice a  week  for  50-weeks.   There was no evidence of squamous
papiilcwas or carcinomas in the mice one year following termination of exposure,
although survival  was  only 35$  (7 of 20).   Doak  et al. (1976) applied estimated
toluene volumes cf 0.05 to 0.1  mi/mouse to the backs of CF1,  C H, and CbaH mice
(approximately  25 mice of each sex of each strain)  twice weekly for 56 weeks, and
failed  tc elicit skin  tumors  or a significantly  increased frequency of systemic
tumors  over untreated controls.  It  is unclear in these studies, however, whether
the toluene  was applied  under an  occlusive  dressing or  allowed to  evaporate.
Li'jinsky and Garcia (1972) did  report a skin papilloma  in 1 mouse and  a  skin
carcinoma in a  second  nouse  in a  group of  30  animals that  were  subjected to
topical applications  of 16 to 20 \ii of toluene twice a week for 72 weeks.
     Frei and Kingsley  (1968) examined the promoting effect of toluene in Swiss
mice following  initiation with 7,12-diraethylbenz[a]anthracene  (DMBA).   In  this
st'irty,  the ears cf the  mine wer° topically  treated once with 0.1 mx, of 1.5? "H3A
in mineral oil  ar:d subsequently,  beginning a week later,  twice  a week with the
same volume  of 10CX toluene  for 20 weeks.   Results showed that 11 of  35  mice
developed tumors  (6 permanent,  5  regressing)  compared witn  8 of 53 negative
controls treated with  100? mineral oil (Table 14-1).  In 14 mice painted  with
100$ toluene but no DMBA  initiator-, 2 developed  tumors (1 permanent,  1 regress-
ing).   In anotner  study with  an identical experimental design, Frei and Stephens
(1968)  similarly found  that 100$ toluene promoted a yield of tumors no different
from that found in  the  concrols (Table  14-1).  In this study,  a total of 7 tumors
were found in  35 surviving mice treated with  toluene following initiation with
DMBA;  th<: negative control  group  (DMBA followed  by biweekly  applications of
mineral  oil) had 3 sKin tumors  in  53 survivors after the  20 weeks.
11.2.   MUTAGENICITY
11.2.1.   Growth Inhibition Tests in Bacteria.  The ability of toluene to induce
DNA damage was evaluated in two  studies  by comparing its differentia?! toxicity to
wild-type and DNA  repair-deficient.bacteria (Fluck et al., 1976; Mortelmans and
                                      1U-2

-------
                                                               TABLE 14-1

                                        Epidermal Tumor Yielc!  in 20 Week Two-Stage Experiments0
DMBA Promoting Age-it
4- None
* 5% croton oil6
* 100J toluene
4- 100? mineral oil
5J croton oil
100J toluene
f 4- None
LO
4- 51 croton oil
4- 100*
4- 5J croton oil
5J croton oil0
100J toluene
No. Surviving
Mice
33b
35b
53b
25b
146
23d
33e
35d
53e
20d
!4d
Tumor
bearing
sifrvivors
NR
NR
NR
NR
NFi
NR
88?
11$
11?
5*
oj
Number of Tumors
Permanent
0
381
6
8
1
1
NR
NR
NR
NR
NR
0
Regressing
0
70
5
0
2
1
NR
NR
NR
NR
NR
0
Total
0
1)51
11
8
3
2
1
352.
7
8
1
0
Tumors
per
Survivor
0
13-7
0.31
0.15
0.11
0.14
0.04
10.7
0.2
0.15
0.05
0
Regressing
Tumors
(J) Reference
0 Frei and Kingsle^ ,
1968
15.5
45.4
0
66.6
5.0
.NR Frei and Stephens,
1968
NR
NR
NR
NR
0
aEars of Swiss mice  treated once with 0.1 mZ of 0.51 DMBA and subsequently, beginning 1 week later,  twice a week with the
.promoting agent.
 Not specifically staged whether this is the number of surviving mice.  Also, the number of mice at  the start not stated.
°In mineral oil
 30 mice at the star',
e60 mice at the stare.
NR = not reported

-------
Riccio,  1980).  Two species were tested with negative results:  Escherichia coli
W3110  (jgol  A*)  ana p3478  (pol A")  and Salmonella  typhimurium SL^525  (rfa)  (ree*)
and SLWO  >fa)  (reef).   In  the  first study,  Fluck  e.t al.   (1976) applied
toluene  (25 ni/plate)  without  metabolic activation  directly  to wells  in  tr.e
center of  culture plates containing the  E.   coli and found  no zones of growth
inhibition with either strain.  In the Mortelmans and Riccio (1960) study, growth
inhibition was  also  found to be   comparable with both the repair* competent  and
deficient  strains of  the E.  coli and S.  typhimurium when sterile  filter  discs
inoculated with O.C01  to 0.01 \i$L  toluene  were  placed in the centers of culture
plates;  these assays were performed both with and without metabolic  activation.
Mortelmans and  Riccio  (I960)  furtner found that  toluene  (0.001  to 0.01  ni/plate)
was not  differentially toxic  to either strain of  the E_. coli or S_. typhimuriun. in
quantitative growth  inhibition tests.  In the  quantitative assays,   the toluene
was preincubatcd in liquid  suspension with the  bacteria,  with and  without  S-9
activation, prior to plating; following plate incubation,  the numbers of surviv-
ing cells  were  counted (instead of recording measurements of diameter.'; of  zones
of growth  inhibition).
T4.2.2.  Tests  for Gene Mutations
     1*4.2.2.1.   ASSAYS USING BACTERIA AND YEAST  — Toluene has been  reported to
be non-mutagenic  in  the.  Ames Salmonella  assay  when tested with strains TA1535,
TA1537,  TA1;.33, TA9S,  and TA100  (Litton  Bionetics,  Inc.,  19?8a; Mortelma.is  and
Riccio,  1930; Nestmann et al., I960; Bos et al., 1981;  Snow et al., 1981), and in
the Z.  coli WP2 reversion  to trp*  prototrophy  assay  (Mortelmans  and Riccio,
1980).  The details  of these studies are summarized  in  Table T4-2.   All assays
were performed in the presence and in the  absence of Aroclor 125^-induced  rat
liver hoaogenate  (3-9) and employed positive and negative controls.   It  should be
noted that  there may  have been significant losses  of toluene from  tne culture
media during incubation  in  all  but one of the aforementioned studies   (Snow
etal.,  1981),  particularly at  the higher doses tested.   Snow  et al. (1981)
conducted  plate incorporation assays in sealed  plastic bags and chambers as well
as vapor exposures  in  desiccators to prevent excessive evaporation.  The design
of the  Snow et al.  (1981) study  is also noteworthy,  because the  toluene  was
tested  wi'.h  toluene-induced rat  liver  S-9 fraction as  well  as  with Aroclor-
induced  S-9.
     Toluene,  with  and  without  metabolic  activation,  was  also  tested  in   IJ.
           for its  ability  to  induce reversions to  isoleucine independence  in

-------
Typ« of Assay
Reverse Mutation
S. typhlRurlua


S. typhleurluo


S. typhlBurlua


S. typhtsajrluB


S. typhlourlua


S. typhlsurlua

f
Ui £. coll

S. c«revlsiae

Kitotic Can* Conversion
S. csrrvUlaa


S. cerovlsl*<8

Kitotic Crossing-Over
S. C-«.-.iVlsi»»

Metabolic
Strain Activation*

TA 98, 100, yes and no
1535, 1537,
1538
Ta98, 100, yes and no
1535, 1537,
1538
TA98, 100, yes and no
1535. 1537,
1538
TA98, 100, yes and no
1535, 1537,
1538
TA98, 100, yes and no
1535, 1537,
15j8
7*98, TA100 re8 *n(1 n°C

yes and noc
V?2 yes and no

D7 y«a «nd no


J.J yes and rs>
°
yes and x>
07 yss and AO


D7 yes ?nd no

ilose Application Ftrspon^c

0.001 to 5.0(il/
plate

0.001 to O.OJII6


0.01 to I0til/plat«


5 Ml/plate


0.115 to ?,~) tit/
pi He

0.3 (il to 100 t't''
plat*
11 to 3761 ppa
0.01 to 10 (it/
pl»te
0.001 to 0.5Jf


0.001 to 5. Out/
pi»t»
0. 138 to 1. t»°
O.C01 to 5.0Jf


O.OOi to 5.0»f


Plate


Llq ild


Plat.


Plato


Plate


Plate

Vapor
Title

Liquid


Plato

Liquid
Liquid


L 1 iju 1 d


Incorporftt 1 on


suspension


Incorporation


1 ncorpor it Ion


Ircorporrit Ion


Incorporation

exposure*
Incorporation

suspension


Incorporation

suspension
suspension


suspension

Hef«renc-«

l.ltlon Blonetlcs, Inc.,
1978a




Horieli&ana and Rlcolo,
19SO

N«!>teann « toluene «j  tea ted  with toluene- IniucJ-J S-9 a.i well as  irlth  »roclor  lndur«d S-9.
"Hhe plates w«re  Incubited In  sealed plaatlc bags or chaisbers  for part  of  a  7?-^r Incut-stlon period; 1n the  Arcclor-lnduced
 S-9 testa, th* plAtca uvrr rrasovei fron thr bags after Ii8 hr,  counlfd,  Incubatfd an ad-tlllon .""I hr, rind recounted;  In tha
 expcrlBCnt^ with  tolu«^*-induced 3-9 th? pl^tpa w*re rcaov»«(J  after  ?**  hr  to prevsnt aolature anil 31-rpaJlng  problffias,  and then
 Incubated an additional  «8 tir Ntfore count Ing.
*Tha aanaya *rrt  run  in  a  »»-*lfl(3 Incubation rh.iist.cr with a .ifrv.nd glass  (ilate (op*n! which contained the toluene;  sftrr
 2*1 hr th* ch^iabers  were open" j and the platps Inrub.iteJ for an additlon.il  $8 hr.
 1001 Borlal.ly al 0.11  an<: O.',l

-------
strain D7 (Mortelsnans and .RicciOi  1980).  mitotic gene conversion  to  tryptophar,
independence in strains DU (Litton Bionetic,  Inc., 197&a) and D7  (Morte.lmans  ana
Riccio,  1980),  and  mitotic  crossing  over  at  the  ade2 locus  in  strain  D7
(Mortelmans and  Riccio.  I960).   Toluene  did not  elicit a positive  mutagenic
response in any of these tests (Table 14-2).
     iu.2.2.2.  TK MUTATION IN L5178Y MOUSE LYMPHDMA  CELLS -- Litton  Bionetics,
Inc. (19?6a) reported that toluene failed  to  induce specific  locus  forward  muta-
tior. in the L5178y Thynidine Kinase (TK)  mouse lytnphoma cell  assay. Toluene  was
tested at concentrations of 0.05  to 0.30  n£/m£.,  with and  without mouse liver  S-9
activation.
1^.2.3.  Tests for Chromosomal Mutation?
     HJ.2.3-1   MICRON'JCLEUS  TEST  IK MICE —  It was  reported  recently by  SRI
International  (Kirkhart,  I960)   that  the  intraperitoneal   administration  of
toluene to male Swiss  sice  failed to cause an increase  in aicronucleated  poly-
chromatophilic  erythrocyte^  in  the  bone  marrow.    Doses  of  250,   500,  and
1000 mg/kg were aizinistered  to groups of 32 mice at 0 and 2U hours, witn sacri-
fices 30,  18,  and 72 hours after the first dose (8 mice/sacrifice).  Five hundred
polychromatic erythocytes per animal were evaluated for  the presence  of micro-
nuclei.  The highest dose tested (1000 mg/kg) approximated the LDj-^  for male mice
(Koga and Ohniya, 1978),
     1^.2.3.2.  XG'J" DOMINANT LETHAL ASSAY — Toluene was reoei/.-ly  evaluated  for
its ability to induce dominant lethal mutations  in sperm  cells of CD-I  male mice
(Litton Bior.etics, Inc,, 1981).  Test mice (12 pen dose) were'exposed via inhala-
tion to targeted exposure levels of  100 ar.d ^00  ppm 6 hours  per day,  5 days  per
week for 6 weeks.  Twelve negative control aice  were  exposed to filtered air in
an identical exoosore regiaen, and 12 positive control nice were injected intra-
peritoneally  with  0.3 mg/kg  triethylenemelaniine  (TEH;  on day *O of  the dosing
schedule.  Following  treatment,  the  males were  mated  sequentially to  2 fenales
per week for  each  of  2 weeks; 1^4 days  after the  midweek  of  mating, each female
waa sacrificed using  COV  and  the number  of  living and dead implantations were
counted.   The  results of  this  study  showed  that tolutne die!  not   cause  ary
significant reduction In the  fertility of the treated Bales, and  did  not  cause
increases in either pre- or post-implantation loss of embryos when  compared with
the negative controls.  A significant induction  of dominant le^r.al  nutations  was
observed in the positive control mice.

-------
     14.2,3.3-   CHROMOSOME ABERRATION  STUDIES -•* Two  reports  from the Russian
literature  concluded that toluene  induced chromosomal aberrations  in  rat bone
marrow cells  following  subcutaneous  injection  (Dobrokhotov,  1972;  Lyapkalo;
1973). In  an analysis of 720 metaphases from the bone marrow of 5 rats th?t had
been subcutaneously injeoted with 0.8 g/kg/day toluene Tor 12 days, Dobrokhotov
(1972) fouqd that 78  (13$) showed aberrations.  .Sixty-six percent of the aberra-
tions were  chromatic breaks,  24$ were chromatid "fractures", 1% were chromosome
"fractures", and 3$ involved multiple aberrations.  The frequency of spontaneous
aberrations in 600  marrow metaphases  from  5 control rats injected with vegetable
oil averaged 4.16$ (65.8$ were  breaks  and 32.4$  were chromatid aberrations;  no
"fractures" or  multiple  injuries were  recorded.).   It was  further found that
similar administration of 0.2 g/kg/day of benzene induced a frequency of  chromo-
somal damage (13.6$)  comparable to  that of 0.8 g/kg/day of toluene,  and that when
a  mixture  of  0.2  g/kg  benzene  and  0.8 g/kg  toluene was  injected daily  for
12 days,  the   damage  was  approximately  additive  (33-33$  aberrations).   The
significance of the positive clastogenic  effects attributed to toluene is dif-
ficult to  assess,  however,  because  the  purity of the  sample  employed was not
stated, and because  the  distinction between chromatid  breaks  and  fractures is
unclear.
     Lyapkalo  (1973) administered  1 g/kg/day  toluene  to  6 rats and 1 g/kg/day
benzene to  8 rats by subcutaneous injection for 12 days.  Treatment with toluene
reportedly  resulted in chromosome aberrations in 11.6$ of the bone marrow cells
examined  (84  aberrant  metaphases/724 cells)  compared  with  3.87$  (40/1033)  in
olive oil  injected controls.   The  types of aberrations that were observed con-
sisted of  "gaps"  (60.47$),  chromatid  breaks  (38.37$) and  isocromatid  breaks
(1.16$).   Benzene caused a greater degree of chromosome damage  than the toluene
(57.2$ of the cells examined had aberrant chromosomes  (573/1002)), and the dis-
tribution  of  aberration types  was different  (44.72$  "gaps",  50.94$ chromatid
breaks^ 4.34$  isochromatid breaks).  The purity of the toluene used  in this study
also was  not stated.
     In a third Russian  study,  Dobrokhotov and Enikeev (1975) reported that rats
exposed  to 80 ppm  (610 mg/m  ) toluene  via inhalation,   4 hours  daily  for
4 months, showed damaged metaphase  chromosomes in 21.6$ of the bone  marrow cells
analyzed*  The percentage of metaphases with damaged chromosomes in bone marrow
cells from air-exposed  control rats was  4.02$.   Inhalation of 162 ppm benzene
caused damage  to chromosomes in  21.56$  of the marrow  cells, and  a mixture  of the

-------
toluene  and benzene  (80 and  162  ppm,  respectively) damaged  chromosomes in ar,
additive  manner (41.21$ of the cells were involved). Chromosome  damage was also
observed  in  all of tne  groups  1 and  2.5 months after the initial  exposure, and  1
lionth after the end  of exposure,  the  frequency  of chromosome damage was still
elevated. A total of 96 rats  were used in this study, but the number of  rats in
each group was not  stated; it should also be emphasized  that  the  number of cells
scored .and the purity of the toluene used were not  reported.
     In   contrast  to the  aforementioned  Russian cytogenetics  studies,  Litton
Bionetics, Inc. (197.8b)  found that  intraperitoneal injection of  pure  toluene
into Charles  River  rats  did  not induce bone marrow  chromosomal  aberrations.
Toluene  was injected  at dose  levels of 22,  71, and 214 mg/kg  in  2 different
experiments.  In 1 study,  5  rats were sacrificed  at 6, 24,  and 48  hours following
injection of each dose; in a second  study, 5 rats were dosed  daily at each level
for  5 days, and the rats were sacrificed 6 hours  after  injection  of the last
dose.  Approximately  50 cells  per   animal  were  scored  for   danage.   Dimethyl
sulphoxide  (DMSC;   the  solvent  vehicle)  administered  intraperitoneally  at
0.65 m£,/rat was used  as a  negative control, and  triethylenemelamine  (TEM) in
saline at  0.3 mg/kg  was used  as  a  positive control.  The  results  of the tone
marrow cytogenetic analyses following  sacrifice are  summarized  in Table 14-3-
It was also noted that  none  of  the observed aberrations differed significantly in
frequency or type  from either concurrent or historical spontaneous values.
     Gerner-Smidt  and  Friedrich  (1978) reported  that toluene at concentrations
of  1.52,  152,  and  1520 ^g/an£ did not influence the  number of  structural  chromo-
somal aberrations  in cultured human lymphocytes.   Benzene and xylene  at the sane
concentrations also  'had  negative  clastogenic   effects  hut  toluene  (152  and
 1520 ug/mZ) and xylene (1520 tig/mil)  caused a significant cell growth inhibition
that was  not  observed  with benzene.   The  data  from this study  cannot  be ade-
quately  evaluated,  however,  because the source and purity of the toluene were not
Stated,  no positive control experiments  were performed, no metabolic activation
system was employed,  an^ the type  of chromosome damage scored was not specified.
     Peripheral blood  lymphocytes of  toluene-exposed rotogravure  workers have
 also  been  examined  for chromosome  aberrations  with negative results.  In one
 study, Forni  and cowotkers  (1971)  examined  the  lymphocyte  chromosomes  from 34
 workers  from a single  plant and  34  controls from outside the plant matched for
 age and  sex.   Ten of the workers were exposed daily  to minimum concentrations of
 131  to  532  ppm benzene  for 2 to 7 years  and  subsequently  to  toluene  in the

-------
                                                                        TABLE  11-3
                                     Rat Bone Marrow Cell  Aberrations Following Intraperitoneal Injection of Toluene
Treatment Dose
DHSO 0.65 ml/rat
(Solvent)


Triethylene 0.3ng/kg
Malaolna

Toluene 22 mg/kg



Toluene 71 ag/kjj



Toluene 211 Bg/kg



Tlae of
Sacrifice
6 h
21 h
18 h
6 h (SA)°
21 h


6 h
21 h
18 h
6 h
6 f
21 h
18 h
6 H
6 h
21 h
16 h
6 h (SA)d
No. of
Animals
5
5
5
5
5


5
5
5
5
5
5
3
5
5
5
5
5
Total No.
of Cells
225
250
250
227
250


250
242
250
238
239
227
150
212
250
250
250
250
Type and Frequency
of Aberrations
Structural
2f,1td

n,b,if
Hd
11tb,23b,5af
26t ,1r,10td.
Ipu, 1qr, 2ac,
	
—
	
3f
ltd
2td,1af,1f
—
—
1f
—
1tb,1td
1td,3af
N'Jraerlcal
__
—
._
--
,l5f, 2pp
12>,
3tr
_.
—
—
--
IPP
—
_-
—
2PP-
1pp
—
--
No. of Cells
With One or More
Aberrations
3 (1.3*)
0 (O.OJ)
2 (0.8*)
1 (0.1J)
72 (28.8*)


0 (0.0*)
0 (O.OJ)
0 (O.OJ)
2 (0.8*)
2 (0.8J)
1 (1.8*)
0 (0.0*)
0 (O.OJ)
3 (1.2J)
1 (0.1*)
2 (0.8*)
2 (0.8*)
No. of Animals
Without
Aterratlons
3
5
1
1
0


5
5
5
3
1
3
3
5
3
1
3
3
Mitotlo
Index8
3.8
6.0
6.1
5.0
1.1


3.1
5.9
7.0
6.3
2.5
1.3
5.7
3.3
1.5
3.6
5.1
5.1
 Source:  Litton Blonetics, Inc., 1978a
 The toluene used was 99.96 wt. J pure (ethylbenzene, 0.03*;  £-xylene,  <0.01*;  m-xylens,  <0.01*;  sulfur, 0.1 ppis) (Fowle, 1981).
°SA s subaoute study; rats were dosed dally for 5 days,  with  sacrifice '6 hours  after  the  last dose
 af = acentric fragment (2 tld); f = fragaents; pp = polyplold;  pu  =  pulverized  chroousoite;  qr =  quadrlradlal; r = ring; sb = chromosoBis break;
 t j translocstion; tb = chromatid break; td = chrcaatld deletion;  tr =  trlradlal;  >  =  greater than 10 aberrations
*Basad on a couit of at least 500 cells per animal

-------
general range of 200 to ^00 ppm for 14 years;  24 of the workers were exposed only
o toluene for 7 t9  15 years,   (the ink solvent used in this plant was changed
from benzene to toluene,  which  contained some xylene, but reportedly no benzene,
after an outbreak  of  benzene poisoning in 1954.)  No significant differences were
found  between  the toluene  and  control  groups  in frequencies  of stable  and
unstable chromosome aberrations or in chromosome counts (Table 14-4).  Approxi-
mately IOC metaphases from each subject or control were scored.  The proportion
of   chromosome   changes  was   significantly   higher   statistically   in  the
benzene/toluene group compared with  controls,  and in  the!  benzene/toluene group
relative to the toluene grcup.
     Maki-Paakkanen  et al.   (1980)  found  no   evidence  of  clastogenicity  in
cultured peripheral  blood lymphocytes from  32 printers and assistants  from 2
different rotoprinting factories  who had  a history of  exposure to pure toluene
(benzene  concentration,   £0.05?;   average  benzene concentration,  0.006?)  at
8-hour, time-weighted average (TWA) concentrations of 7  to  112 ppm.  The average
age  of the workers  was  34.2 years  and  the average  length  of  employment  was
•14.6 years.   Results of  analyses showed  that when frequencies  of  chromosome
aberrations were  compared with those of  15  unexposed research institute workers,
there  were no significant differences (Table 14-5).  Similarly,  no significant
deviation?  were  observed  in  the frequencies  of - aberrations  in relation  to
duration of exposure.
     Bauchinger  et  al.  (1982)  performed cytogenetic  analyses  on  peripheral
lymphocytes from 20 male rotogravure  plant workers who were  exposed for _>16 years
to  toluene  that contained 
-------
                                                          TABLE 14-4

                              Frequency of Unstable and Stable Chromosome Changes and Chromosome
                                   Counts  in  Subjects  Exposed  to Benzene  or Toluene  or Both

Expsoure

Benzene
Toluene
Control

Subjects

( + toluene)

subjects
No. of
Cases

10
24
34
Age
Range

36-54
29-60
25-60
Total
Cells
Counted

964
2,400
3,262


C
u
1.66(1
0.80(0
0.61(0
% Cells

b

<87)d,e,f
.83)d
.67)


C
s
0.
0.
0.
% Cells

Q

62e'f 13.1
08 14.3
09 10.2
With Chromosome Number

46

86
85
89



.0
.4
.5

>U6
(Polyploid)
0.9(0.52>
0.3(0.29)
0.3(0.3)
 Source:   Forni et al., 1971
 Cells with "unstable" chromosome aberrations (fragments,  dicentrics,  ring  chromosomes).  The presence of each
 fragment was considered as one break,  the presence  of a dicentric  or  ring  chromosome as two breaks.
 Cells with "stable" chromosome changes (abnormal  monocentric  chromosomes due to deletions, translocations. etc.,
 trisoraies)
j
 Numbers  in parentheses show percentage of calculated  breaks.
 Difference from toluene group was significant (P  <  0.05)
f
 Difference from control was significant (P < 0.01)

-------
                                                                        TABLk.  11-5
                      EfCecl. of Occupational Toluere Exposure £.nd Smoking on Ci.romoaomal Aberrations and Slater Chromatld Exchanges
Cells with Chromosomal Aberrations (1)










— •
t
r\>








Occupational
Toluene Exposure
(yr)
Total Worker
(11.6 yr average
Total Cortrol
C (controls)
Nonsmokers
Smokers
Total

1-10 Voean, 8.0)
Nonauoker;)
Snokera
Total
>10 Joean, 19.3)
Norafflokers
Sookers
lotal


No. of
Subjects
32
exposure)
15

M
11
15


3
to
13

1 *
8
19

Mean
Age
(yr)
31. 2e

31. 2e

31.0
3<; _ 5
31.3


27.7
28.2
28.1

38.5
35.9
37.5


Cells
Analyzed
,._

—

800
1100
1900


300
1000
1JOO

1100
800
1900


Chroma t Id
Type
1.0

0.7

0.5
0.9
0.7


0.7
0.7
0.7

0,8
1.8
1.2
Gaj,).° Excluded

Chromosome
Type
0.5

0.9

0.8
1.0
0.9


0.3
0.3
0.3

0.5
0.8
0.6
Sister ChromatM Exchanges (SCEs)


Total
1.5

1.6

1.3
1.8
1.6


1.0
1.0
1.0

1.1
2.5
1.8

Gaps Included
Total
2.5

2.7

2.3
3.1
2.7


2.3
1.9
2.0

2.5
3-1
2.8

Cells
Analyzed
	

—

231
318
552


79
295
371

330
205
535

Hear per Sub
per Cell
8.5

8.9

8.0
9.7"
9.2


7.9
9.1"
6.8

7.5
9.6»*
8.3

iect











9



•

 Source:  Makl-Paakkanen  et  al.,  1980
 100 cells analyzed  per Individual
°30 cells analyzed per individual
 Calculated fron  Individual  roeana
 hean value
f.--               ,  r
 OX'OJ Mci'o ^Uj  f oU
yr i year
*"F '. 0.0' and •*• P '. C.001 ~:-parci tc norajiokcrs In  the group,  one-tPlleJ  Student's t-test

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                                                                       TABLE 11-6
                                    Mean Frequency of Chromosome Abnormalities * S.E. a'id SCEs + S.E. In L
                                            of Toluene- Exposed Rotogravure Workers and Unexposed Controls'^
ytnphocyte
No. of
Subjects
Toluene-Exposed 20°
Unexposed Controls 21
.
Age
(years)
Caps
S Cells6
I.S.D. per cell (J)
'f'4.2 » 0
7.0 ~ 0
12.1 * 0
10.5 0
.0218 +
.0021
.019 t
.003
0.90 t
0.13s"
0.51 +
0.06
Aberrations per Cell
Chromatld
breaks
0.0036 +
O.OOIO6"
0.0019 +
0.0005
Chromatid
exchanges
0.
0.
0.
0.
0015-+
0005
OOOU +
0002
Acentric
fragments
0
0
0
0
.0035 +
.0008
.0023 +
.OOOll


Dicentrlca
0,
0.
0.
0.
.0005 +
,0003
.0005 +
.0002
SCE
per cell
9.62 +
0.37^
6.18 +
0.25
"source:   Bauchlnger et •!.,  1982
 300 cells and 500 cells were analyzed for  structural  chronoscmal  cbangea  In each exposed and control subject, respectively.  Fifty cells/subject were
 scored for SCEs.
C11 heavy snokerj (>10 clfiarettes/day),  1 moderate  snoker, and  8 non-snokers
 8 heavy smokers, 1 moderate  aooker,  and 15 f^on-smokers
 cells with structural chromosome changes  (S-oells)
 Difference f. as the unaxposed control group was  significant  (P<0.05-)
Siffwenos free the unexposed control group was  significant  (F<_0.0!)
 The subjects of either group were subdivided Into  smokers and  non-saokers for statistical evaluation.  Non-smoking workers had significantly higher
 (P=0.02) SCE values (8.55 *  0.27) than  non-smoiclng controls  (7.75 + 0.25), fi)id smoking workers lad significantly higher (P=0.020) SCE values (10.33
 t O.t9)  than aaoking controls (8,85  + O.'JI).

-------
 cantly  higher.   Although the  workers examined  in this  study were  exposed to
.levels  of  toluene  that, are similar to those reported  by Forni  et al,   (1971) and
 Maki-Paakkanen  et  al . (I960),  it .should  be  noted  that a,-greater  number of
 cells/individual we^e  scored for aberrations  ("300  or  vDQ versua 100).
     J'R- a  report  or,  chromosome  aberrations   or"  women i-n  laboratory  work,
 Funes-rCravicta  et  ai.  C197Y) «*lsj presented datH'.on T< workers who were exposed
 to  toluene  in a rotogravure factory.  Exposures  ranged from 1.5 to 26 years and
 air measurements  of toluene show-ed TWA values of 100 to 200 ppm, with occasional
 rises up tc 500 to 700  ppa; tr,t exposures were sufficient  in cnotst cases to elicit
 frequent he'acjacr.^s ar:d fatigue, and occasional  vertigo,  nausea, and feelings of
 drunkenness.  T.-.e  workers had teen exposed  to  toluene since approximately 1950;
 before  195i,  ir- way at,at-;d t.'r.ai. toe toluene was probaoly  contaminated  by a "low"
 percentage of  ber.:".:r.i.    Hesults  of  lymphocyte  analysis showed  an  excess of
 chromosome aierrati^r.s  (ahr,jr~ui  chromosomes  and breaks)  in the  14 toluene-
 exposed workt?rs relative to a control group of  1(2 adults.   It should be noted,
      i
 however, that only a saiiii;  r.umber  or'  subjects  were examined in  this study and the
 exposure badure re  toluene at concentrations  Of  15.2, 152,  and
 1520  ug/nii. hac r;c  <=-rffcct  oi; '.se  number of  sist^r-chromatid  exchanges (SCEs) in
 culturea hur-an  1-,-Trspn...', ytes,  but no  positive control  experiments  were performed
.and no- meta^ii^'  activati.'.". syot.em  was  eraployed.   Twenty-six  cel.\s/dose were
 scored  fcr iCJ^r and cyt^toxi': i ty  was  observed  at  the highest  dose.   Evans and
 Mitchell {'i9 112 ppm
 pure  toluene, Maki-1-aakkanen et al. (1980) found no increase in SCEs relative to
 a  group of "5 unexposed  control  subjects.  The  average  age of  the worksrs was
 31.2 years and  their  average  length of  employment   was 1^.6 years.   The SCE

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                                 TABLE  11-7
           Chromosome Aberrations in Rotoprinting Factory Workers

No. of Subjects
Age (year)
Ra.'.ge
Mean
No. of Cells Analyzed
Total
Aononnal
Total
Frequency range (J)
Mean frequency ($)
No. of Chroaosomes Analyzed
Total
Breaks
Total
Range (per 100 cells)'
Mean (per 100 cells)

Control
H9

0.16 to 63
21. 1

5000

217
0 to 20
1.3

230,000

233
0 to 22
5.1
Group
Toluene
11

23 to 51
37.2

1,100

108
2 to 15
7.7

61,100

121
2 to 17
8.9


Benzene/Toluene
8

51 to
61.3

800

76
1 to
9.5

36,800

95
6 to
11.9


65





17





17

Source:  Funes-Craviota et al.,  1977
Exposure details provided in accompanying  text.
                                     11-15

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analysis was part of a study examining chromosomal aberrations in these workers;
the exposure history of the subjects is described in more detail with the summary
of the aberration  findings (Section  11.2.1.1.),  and  the.results  of the  SCE
analyses  are  included  in  Table 11-5.
     Bauchinger et  al.  (1982)  reported a significantly increased number of SCEs
per peripheral  lymphocyte in a group of 20 male rotogravure workers who had been
exposed to 200-300  ppm  pure  (<0.3?)  toluene for >l6  years  (Table 11-6).   As
described  in  the more  detailed Section  11.2.1.1 sumn00  to  700  ppm), but benzene  concentrations were riot
measured.   The technicians also  had a  history of exposure to toluene,  but the
exposures  were poorly  characterized (duration and concentrations  not stated) and
each had considerable concurrent exposure to other solvents as  well, particu-
larly  benzene  and  chloroform.    Results  of peripheral  lymphocyte  analysis
(20 cells/individual scored) showed a statistically significant increase in SCEs
in the laboratory  technicians  and the  children of female technicians,  but not in
the exposed printers;  however, due to the nature of the exposure, the increases
noted cannot  be exclusively attributed to toluene.
11.3-  TERATOGENICITY
11.3.1.   Animal Studies.   Toluene  was  reported  in a recent   abstract  to  be
teratogenic to  CD-1 mice following oral  exposure  (Nawrot and Staples,  1979).
Toluene was administered  by gavage from days  6-15 of gestation at levels of 0.3,
0.5, and  1.0 m£/kg/day  (approximately 0.26,  0.13,  and  0.87 g/kg/day,  respec-
tively) and from days  12  to 15 at  1.0 mil/kg/day. The vehicle used was cottonseed
oil (0.5$ of maternal body weight  per dose).   A significant increase in embryonic
lethality occurred at all dose levels  when  administered on days  6 to 15, and a
significant reduction in fetal  weight  was  measured in the  0,5  and  1.0 mi/kg
groups.   Exposure to  1.0 mJZ./kg   toluene  on  days  6 to 15 also significantly
increased  the incidence of cleft palate; this  effect reportedly did not appear to
be due merely  to a general retardation in growth rate. When toluene was adminis-
tered at  1.0 ra£/kg  on days  12 to  15, however, decreased maternal  weignt gain was
                                     11-16

-------
the only effect observed.   Maternal toxicity was  not noted after  exposure to
toluene on days 6  to  15 at any  dose  level.   It should  be  emphasized that the
numbers of mice exposed and the numbers of fetuses  examined  were not stated in
the available  abstract  of this  study;  a  complete  copy  of  this  report  is not
available for review but has been submitted for publication.
     Hudak and Ungvary  (1978)  recently concluded that toluene  was  not terato-
gen\ic to CFLP mice  or CFY rats when administered via inhalation according to the
following schedule:
                       Dose
       CFPL .nice
133 ppm (500 rag/m3)
399 pps (1500 mg/in3)
       CFY rats      266 ppc (1000 ag/m3)
                     399 ppm ( 1500 mg/nr )
                     399 ppm (1500 mg/m3)
Days of Pregnancy
      6-13
      6-13

      1-21
      1-8
      9-1M
Duration
2k hours/day
2*1 hours/day

 6 hours/day
2^4 hours/day
2*4 hours/day
It was found that the entire group of mice exposed to  399  ppm toluene died within
2*4 hours.  Toluene administered to rats  at 399 ppm also had an  effect on maternal
survival, but  none of  the exposures adversely affected the incidence of external
or visceral  malformations,  in  either species relative  to  air-exposed controls
(Table 14-8).  An  increased incidence of skeletal anomalies  (fused ste^nebrae,
extra ribs) was observed, however,  in the rats exposed continuously to 399 ppm
tcluwie on days 9  to  1*4.  and  signs of retarded skeletal development (including
poorly ossified s'cernebrae, bipartite vertebra centra, and shortened 13th ribs)
were found in the  rats  exposed on days 1 to 8  (399  ppm)  and  during the entire
period of  pregnancy (days  1 to 21) at 266 ppm for 8 hours/day.  An erobryotoxic
effect of toluene was  further indicated  by low fetal  weights in the mice, and in
the rats  exposed  on  days  1 to 8  of pregnancy.  Fetal  loss  (percent of total
implants), mean litter size, mean placental weight, and maternal weight gain were
unaffected by exposure in either species.
     In a more recent  teratogenicity study, groups of 20 CFY rats were exposed to
266 ppm (1000 mg/m )  toluene,  125  ppm WO mg/nr) benzene,  or a combination of
these concentrations of  toluene and  benzene vapor for 2*4 hours/day on days 7 to
H of gestation  (Tatrai  et al., 1979).   A  group of 22  rats  inhaling  pure air

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                          TABLE  11-8




Toralogenlclty Fvaluallr/n of Toluene in Cfl Rats and CP1P Mice*





No. pregnant anloals exaalned
No. pregnant an! Mia died
Maternal weight g*lnb (J)
No. live fetuses
No. resortxd fetuses
c: No. dead fetuses
— Fetal loss (J)
Mean litter site
Mean fetal velght (g)
Mean placantal weight (g)
Weight retarded fetuses0 (J)
External oal format lorj
No. fetuses dissected
Internal ealforaatjona
Anophthalaia
Hydrocephalua
Hydronephoroa 1 -.
No. of Alizarin-stained
frtuses
Skeletal retardation signs'

Air Inhalation

Days 1 to ?1 t
8 h/d
10
0
16.6
111
8
0
6.7
11.1
3.8
0.5
7.2
0
51

0
--
1

57
0


?M> ppa
JllfS 1 tO
8 h/d
10
0
11.1
133
3
0
2.2
13-3
3-6
0.5
16
0
61

0
..
6

69
17"

Toluene
399 ppo
21 flay.i 1 13
21 h/d
9
5
11.0
95
6
0
5.9
10.6
3.3"
0.5
16"
0
19

0
1
H

12
7"
Flats
Air Inhalation

B Days 9 to 11
21 h/d
?6
0
16.9
318
15
0
1.1
13-1
3.6
0.5
6.9
0
;79

1
--
16

169
11

Toi uene
399 ppo
Days 9 to 11
21 h/d
19
?
11.8
213
1B
0
7.8
11.2
3.8
0.5
17.3
0
110

0
—
4

102
21"

Kir Inhalation

Days 6 to 13
?1 h/d
11
0
—
121
6
1
6.1
9.0
1.1
--
6.5
0
6!)

0
—
1

60
3
Mice

Toluene
133 ppo
Days 6 to '3
21 h/d
11
0
—
112
10
0
8.2
10.2
1.0»
--
27.6"
0
58

0
--
3

51
1
399 ppa
Days 6 to 13
21 h/d
0
15

0
0
0
0
--
~
—
—
__
0

—
—
«

—
-_

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                                                                TABLE 14-8 (cont. )










_,
tr
1





Skele'.al anomalies
Fused stsrnebrae
Extra ribs
SkeH-lal malformations'1
Missing vertebrae
Brachizella



Air Jnl a! at lor,

Daya 1 to 21
8 h/d

3
0

0
0



Toluene
2bt ppns 399 ppn
lays 1 to ?' Day« ! tr
8 h/d 2!i l./d

0 .0
0 0

0 0
0 0


Rats
Mr Inhibition Tolunne
39') ppffi
B rays 'i to 14 Days 9 to
tJU h/J 21 h/d

2 ?••
0 2?'*«

0 2
- 0 0


Mice
Air Inhul stlon Toluene
133 ppra 399 Ppo
11 Dnys 6 to 13 Daya 6 to 13 Days 6 to 13
24 h/d 24 h/d 24 h/fi

0 0
0 0 —

0 0 —
1 0


a
 Source:  Hudsk and Ungvary,  1978
 Percent of starting body weight
°Porcent of living fstuaea weighing <3.3 g (rats) or 0.9 g (sice)
 Agnathla, brachloclla, tlaalng ti.ll
 The rats were sacrificed on day ;;l of pregnancy, the nice on day  18
 Thynus hypolasla alao looked for
 Including poorly ossified stsrnebrae, bipartite vertebra centra,  and rmortened Ijih  ribs
 rissura aternl and agnathla alsd looked for
•P < 0.01 (t-test); •« P < 0.05  :Hann Whitney U Test); ••• P <  0.01  (K»nn Whitney  U Teal)
t, i hour; d = day

-------
served as controls,  and  trw fefj.-s^s  were exa.tin*.",! t^ri ciay .^ *  of pregnancy.   The
results of the toluene exposures ir,-Mus study are consistent  wit.r: those ot  l-udak
.and Ungvary's continuous  TV-* p.;'" t.Jje-ie ex^csjre^ with rats  • or -Jays  9  to  lit of
gestation.  ?atr*i  et a!.   '^"'.S  c-j:.cl'j-.i';J -tn-:it : or.li r.uc.ia' t-xpiiurt  to 266 ppa
toluene was  not teratoger.se (no • exterr.ai ,  internal,  or: skelct ji aalforaaticns
.were reported), aithougn  the e.<;osu: es were dssiciatea with »/i.-:er.ce of  skeletal
retardation   (not   detailed)    ar;:   ar:  increased  insidenvt  of   extra   ribs
(Table 1^-9) •   It  was additionally-  fcunc tnat  tr.e incidence of  extra  ribs was
higher in the group  exposed f;  toluene  in ?oa;tir.4t:-jn  with teiz^rit* than  ir. the
groups exposed  to toluene alor.e.  Mat^r.'.ui less, m;»t^rr.al weight  gau., n-icber of
litters, aear.  ispla.ntatiori'aaa, plaoer.tal we^^rit, fetal loss,  arid fetal  -eight
less  were r.c-t  s..5r.i!':".a:*  y  >: r--..-ted  by t: e  toluene  exposures.   Exposure to
125 PPfB bentene -JAd cause  jecrre liies in aaterr.-j;  weight pain, placer;tal  weigr.t and
fetal weight,  but these  effects app-ear-d *-   ce  :r.n:bited by concurrent  exposure
to 266 'pps toluene.   Further, it war.  reported tr.at po'st-iapiar.tation  fetal  loss
(the nucber  of  dead and  resort;: f^t-jses relative to the nuaber wf  total implan-
tation sites in percent'  wa? sigr.iiic2~.tiy  increased  in  the;  grsup  exposed to
ber.zer.e  in -v.^Dinatior. w: tn tolu»T:r;  feta.  loss ua.s  r.ct,  as  indicated  earlier,.
affected  by  exposure to  tne t^l_ehe  (or b-rri-.i.-ie;  alone.
     In  a -third .inhalati'jr study, "jitt-:,  £:or.-:tlca.  Inc. (1972C-J rtpcrted no
evidence cf  teratogeni .: u/  in tr.*-  :•)-,^y-.'-c f?--. :3ti  of Cnarles Piver rats  that
were exposed to 'Cj or w >'.•  pps:  toluene  ;apr,»- for  r; :;ojr3/day  on day^  c  to  15 of
gestation.   Histologi.;al rx:: .rS r.acior.a revea^e..;  uo unusual  incidence of  visceral
or  skeletal  ahr.r -rs.al ities  ;7atle  " • - ' f -  ;   oinu.ijal   skeletal  variation;,  were
observed  in'  a  scall  out  cotipar^oie .-,-._;o>-:r -if !etu.?^i from  both the exposed ana
control groups,  but  tr.^ae c-! .t.i^es  «-re  iri -ost  c^es  attributed to  retarded  bone
ossification  arid were r.ot cc/.-.^idL-T-'.-'i t>, fc-t  U3^fonr.a';ior
-------
                      TABLE  14-9

Teratc-^enic  Effects  of  Exposure to Toluene, Benzene^
arid a ^cunbir.ation of Toluene and Benzene  in  CFY  Rats"
Inhalation or. days
7 to 14 of pregnancy
2-K h/d
Hunter of females
treated
dies
r.or. pregnant
total rescrption
Nusbtr cf liters
Hear, ixplantation/caa
Maternal weight gair.
in i of starting body
weigh'.
Re'.itive liver weight
U}'
Hear, placenta! weight
(f,'>
i
Naaber of fe-tuses
live
dead
resarbed
Mean fetal weignt CgJ

Weight retarded
fetuses in i of .living
fetuses
External malformations
Fetal loss/ total
Implantation sites (%}

-------
                                    TABLE 14-9  (cont.)
Inhalation on days
1 to 1A of pregnancy
24 h/d
Skeletal anomalies
sternua m-isal igned
asyaaietric vertebra
extra ribs

Air Toluene
266 ppm

h h
1
1 7*

Benzene
125 ppc

5
3
1

Toluene/Benzene
266 ppm •* 125 ppo

1
1
19*"
oigrdf icance
of
Interaction




Skeletal malformations
No.  fetuses dissected
Internal nalformations
  poiycystic lungs
  pyeiectasia
  dystopia renic
  vesica giganta
  nicrophthalmia
  ar.opjithalaia
 •hyflrocephalus
   ir.ternus
133
118
116
"Source:  Tatrai  et  al.,  1980

+ --  p < 0.1; *  =  p < 0.05; ••  -  p 
-------
                                  TABLE 14-10
                  Teratogenicity and Reproductive Performance
                    Evaluation  in  Rats Exposed  to Toluene
v. .
Pregnancy ratio
(Pregnant/Bred)
No. pregnant rats that- died
Live litters
Implantation sites
(Left Horn/Right Horn)
Resorptions
Litters with resorptions
Dead fetuses
Litters 'with dead fetuses
Live fetuses/implantation site
Mean live litter size (fetuses)
Average fetal weight (g)
Number of fetuses examine for soft
tissue (visceral) changes^
Nuaber of fetuses examined for
skeletal changes
Number of fetuses with normal
skeletal examinations
Fetuses with commonly encountered
e f
skeletal changes '
Fetuses wij,h unusual skeletal
variations' 'g

0
26/27
0
26
152/194
26
13
0
0
320/3^6
12
3-6

108(51/57)

212

139
67(20)
6(4)
Dose (ppm)
100
27/27
0
27
181/177
28
20
1
1
329/358
12
3-5

105(47/58)

221

150
62(20)
9(4)

400
27/27
0
26
179/190
4lb
17
0
0
328/369-
12
3-8

104(51/53)

224

158
58(20)
8(6)
 Source:  Litton Bionetics, Inc.,  1978b
 The increase in total resorptions at this dose was attributed  to  the total
 resorption of the litter of one particular female.
Cumbers of male/females examined  in parentheses.
Tour specimens froa one litter were not examined  (missing).
 A  qualitative examination of the  observations recorded  for the fetuses  indicates
 that bilateral ribs, unilateral ribs, and reduced ossification of various bones
j.were the most frequently encountered changes.
 Number of litters in parenthesis.
T'hese were generally cases of more severe and extensive retarded  ossification.

-------
incubation.   Survival incidence  after  14  days of  incubation  appeared to  be
influenced only after injection of toluene on day 6 at 100 umol/egg;  the "approx-
imate LE>  " for  toluene was  judged to  be  in excess of 100 umcl/egg.   Macroscopic
examination on day 14 indicated that only 3 of 46 of the  chick  embryos  treated
with 5 to 100 |imol/egg of  toluene  were malformed; 1 displayed profound edema and
3 had skeletal abnormalities (musculoskeletal  defects of the lower  extremities,
but not wings).
     Mclaughlin et al. (1964) injected toluene at dose  levels of 4.3,  8.7,  and
17.'4 mg  into  the  yolk sac  of fresh  fertile  chicken  eggs   before  incubation.
Following incubation, the  percentages  of  hatch at the  3 doses were, respectively,
&5%, 25?, and 0%.  Teratogenic effects were not observed in either the eggs  that
failed to hatch or in the  chicks  that, did hatch.
14,3-2.   Human Reports.   Holmberg  (1979)  gathered  information  on  exposure  to
noxious agents during the  pregnancies  of  120 mothers of children  with congenital
CMS defects and tneir matched-pair controls.   The matched-control mother is the
mother whose  delivery immediately  preceded that  of  the  case mother in the  same
Finnish maternity welfare district.  Results showed that  14 of  the 120  case
mothers  had  been  exposed more  often than  control  mothers   (3/120)  to  organic
solvents during the  first  trimester of pregnancy.  Among the  14 exposed mothers,
2 had been exposed to toluene.  One of thf toluent-exposed mothers  (age  18)  had
reportedly been exposed in  the metal  products manufacturing industry (no other
details of exposure  given),  and gave birth to a child that died after  2 hours and
showed internal congenital hydrocephaly and agenesis  of the corpus  callosum  upon
autopsy;  other findings  included pulmonary  hypoplasia and  a diaphragmatic
hernia.  The  other mother was exposed to toluene  concomitantly  with other  sol-
vents  (xylene, white spirit, methyl  ethyl  ketone) during rubber products manu-
facturing; her child was  hydranencephalic and died 24 days after birth,   it was
noted that in this case parental age (maternal, 42 years;  paternal,  44 years) and
a previous child with brain injury (born 20 years  previously,  died at age 4)  were
more likely than the recent  exposure to have predisposed the  more recent child to
the defect.
     Toutant  and  Lippman  (1979)  described  the  birth  of a  child  with  "nearly
classic" fetal alcohol syndrome to a 20-year-old primigravida whose major addic-
tion was  to solvents (reportedly, primarily toluene).  This  woman  had a 14-year
history of daily heavy solvent abuse  (no details provided) and a 3-year history
of  alcohol intake  of about  a six-pack of  beer  weekly.   On  admission,  she
                                     14-24

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exhibited signs compatible  with severe solvent  and/or alcohol  abuse  (ataxia,
resting and intention tremors,  mild diffuse sensory deficits, short-term memory
loss,  and poor intellectual  functioning).  The child was born at term, was small
(10th percentile in weight,  5th percentile in head size), and exhibited abnormal
features that included microcephaly, a flat nasal bridge,, hypoplastic mandible,
short palpebral fissures,  mildly low-set ears, pronounced sacral dimple, sloping
forehead, and uncoordinated arm movements.  It was  noted  that although solvent
abbse rather  than  alcohol predominated in  this mother's addiction pattern,  the
case seemed no different from reports of fetal alcohol syndrome.
14.1.  SUMMARY
     CUT  (1980)  concluded  that  exposure  to 30,  100, or  300  ppm  toluene  for
24 months did  not  produce an increased incidence of neoplastic, proliferatiye,
inflammatory,  or  degenerative  lesions in rats relative to unexposed controls;
the highest  dose  tested  was  not,  however,  a maximum  tolerated dose.   Other
studies indicate that toluene is not carcinogenic when applied topically to the
shaved skin of mice (Pohl,  1973;  Lijinsky and Garcia,  1972; Coombs et al., 1973;
Doak  et  al.,  1976),  and that it  does not  promote the development of epidermal
tumors  following  initiation  with  PMBA   (Frei   and  Kingsley,  1968;  .Frei  and
Stephens, 1968).
     Toluene  has yielded  negative  results  in  a  battery of microbial, mammalian
cell, and whole organism  test  systems.  The microbial assays conducted include
differential toxicity testing with wild-type and  DNA  repair-deficient strains of
E.- coli- and f3.  typhimurium (Fluck et al.,  1976; Mortelmans and Riccio, 1980),
reverse mutation testing with various strains of S. typhimurium, E. coli WP2, and
S. cerevisiae D7  (Litton Bionetics, Inc.,  1978a; Mortelmans  and Riccio, 1980;
Nestman et al., 1980), and mitotic  gene conversion and crossing-over evaluation
in 5, cerevisiae D4 and D? (Litton Bionetics, Inc.,  1978a; Mortelmans and Riccio,
1980).   Toluene also failed  to induce specific  locus  forward mutation in the
L5178Y  Thymidine   Kinase  mouse lymphoma  cell  assay  (Litton  Bionetics,  Inc.,
1978a), was negative in the micronucleus test in mice  (Kirkhart, 1980), and was
negative  in  the mouse  dominant lethal assay  (Litton  Bionetics,  Inc.,  1981).
Sister-chromatid exchange (SCE) frequencies were not  altered in Chinese hamster
ovary cells (Evans and Mitchell, 1980) or in human lymphocytes  (Gerner-Smidt and
Friedrich,  1978)   cultured  with toluene _in vitro.    The  frequency  of  SCEs in
peripheral lymphocytes  from workers that  had a  history of chronic  exposure to
similar levels of  toluene has been  reported to be increased  (Bauchinger et al.,
                                     14-25

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1982)  as well as uncharged (Funes-Craviota et al., 1977; Maki-Paakkanen et al.,
1980).
     In the Russian literature,  chromosome aberrations  were reported in the bone
marrow cells of rats exposed subcutaneously (Dobrokhoto'f,  1972; Lyapkalo, 1973)
2nd via inhalation  (Dobrokhotov and Enikeev,  197f )  to toluene.  These findings
were not corroborated, however, in a Litton Bionetics, Inc. (1978b) study in rats
following intraperitoneal injection or in cultured human .lymphocytes exposed to
toluene in vitro  (Gerner-Smidt  and  Friedrich, 1978).   An excess  of chromosome
aberrations in lymphocytes free workers who were chronically exposed to similar
levels of toluene  has been reported by Bauchinger  et al., 19£0 and Funes-Craviota
et al., 1977, but not by Forni et al.. '971 or Maki-Paakanen  et al., 1980; it is
probable, however, thai  part of  the  exposure in the Tunes-Craviola et al. (1977)
study was to benzene-contaminated toluene.
     Toluene was reported in a recent  abstract from NIEHS to induce cleft palates
at a level of 1.0  mJl/kg  (0.87 g/kg)  following  oral exposure to mice on days 6 to
15  of  gestation  (Nawrot and  Staples,  1979);  significant increases  in embryo-
lethality and decreases  in  fetal  weight  were  noted as well at doses  as  low as
0.3 m/kg/day and  0.5 m/kg/day, respectively.  The teratogenjc effect reportedly
did not appear to  be due merely to the general  retardation in growt'i rate.  Three
other  studies  concluded  that toluene  is not tpratogenic  in mice  (Hudak  and
Ungvary,  1978) or rats (Hudak and Ungvary, 1978;  Litton B:onetics, Inc., 1978b;
Tatrai  et al.,   1980)  following  inhalation   exposure.   Embryotoxic  effects
(increased  incidence of  skeletal  anomalies  and  signs  of  retarded  skeletal
development,  low  fetal   weights)  and  increased maternal mortality  wer« noted,
however,  in  some  of the  rats and mice  exposed  via inhalation.    Injection o*"
toluene  into the  yolk  sac  (McLaughlin   et al.,   1964)  or air space  (Elovaara
et al., 19795) of chicken eggs before incubation or during development, respec-
tively, did not result in teratogenic effects.
 14.5  REFERENCES

 BAUCHINGER,  M.,  SCHMID,  E.,  DRESP,  J.,  KOLIN-GERRESHEIM,  J.;  HAUF,  R.  and
 SUHR, E.  (1982).  Chromosome changes  in  lymphocytes after occupational exposure
 to toluene.  Mutat.  Res.   102(4):
                                     14-26

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BOS, R.P.; BROUNS, R.H.E.; VAN DOORN,  R.;  THEUWS,  J.L.G.;  and HENDERSON, P.T.
(1981).   Non-mutagehicity of  toluene,  p-,  m-. and  p-xylene> o-methylbenzyl
alcohol  and  ^-methylbenzyl  sulfate   in  the  Ames   assay.     Hutat.  Res.
88^0:273-279.

CHEMICAL INDUSTRY INSTITUTE OF TOXICOLOGY (CUT).   (I960).  A  twenty-four month
inhalation toxicology study in Fischer-S'J1! rats  exposed  to  atmospheric toluene.
Executive  Summary   and   Data  Tables.    Conducted  by   IndustriaJ  Bio-Test
Laboratories, Inc., Decatur,  IL,  and Experimental Pathology Laboratories, Inc.,
Raleigh, NC, for CUT, Research Triangle Park,  NC.   October 15,  1980.

COLEMAN, G.L.,  BARTHOLD, S.W.,  OSBALDISTON, G.W., FOSTER,  S.J. and  JONES, A.M.
(1977).   Pathological changes during aging in barrier-reared Fischer 3^ male
rats.  ±. Gerontology.  32: 258-278.
    I
COOMBS, M.M.; SHATT,  T.S.;  and CROFT, C.J.  (1973).  Correlation between carcino-
genicity and chemical structure in cyclopenta[a]phenanthrenes.   Cancer Research.
33:832-837.

DOBROKHOTOV, V.B.,  and  ENIKEEV,  M.I.    (1975).  Mutagenic effect  of benzene,
toluene,  and a  mixture  of these hydrocarbons  in  a chronic  experiment.   Gig.
Sanit.   J_:32-3^.   (In  Russian  with English summary;  evaluation  based  on an
English translation provided by  the U.S. EPA.)

DOBROKHOTOV, V.B.  (1972). The mutagenic influence of benzene and  toluene under
experimental conditions.  Gig. Sanit.  ^7_:36-39.   (In  Russian;  evaluation based
on  an English translation provided  by the U.S.  EPA.)

DOAK, S.M.A. et  al.   (1976).  The carcinogenic response in mice to  the topical
application of propane sultone  to the skin.  Toxicology. 6^:139.
                                     1U-27

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ELOVAARA,  c).  et al.   (1979).   Effects of methylene  chloride,  trichloroethane,
trichloroethylene,  tetrachloroethylene and toluene on  the development of chick
embryos.   Toxicology.   12(2):111.

EVANS,  E.L.,  and-MITCHELL,  A.D.   (1980).  An Evaluation of the Effect of Toluene
on Sister  Chromatid Exchange  Frequencies in  Cultured  Chinese Hamster  Ovary
Cells.    Prepared  by  SRI  International, Menlo  Park,  CA,  under Contract  No.
68-02-29^7 for  the U.S. Environmental Protection Agency, Research Triangle Park,
NC,

FORNI,  A.; PACIFICO, E.;  and.LIMONTA, A.   (197').  Chromosome studies in workers
exposed to benzene or toluene or both.   Arch.  Environ.  Health.  22(3):373-378.

FLUCK,  E.R. et  al.  (1976).  Evaluation of a DNA polyraerase-deficient mutant of
E.  coli for the rapid detection of carcinogens.  Chem. Biol. Inter.  15:219.

FREI, ,J.V.,  and KINGSLEY, W.F.    (1968).   Observations  on  chemically induced
regressing tumors of mouse epidermis.   J_. Natl. Cancer. Inst.  J4J: 1307-1313.

FREI, J.V.,  and STEPHENS,  P.   (1966).   The correlation  of  promotion of tumor
growth and of  induction of hyperplasia  in  epidermal  two-stage carcnogenesis.
BrU. ,;. Cancer.  22:63-92.

FUNES-CRAVIOTA, F. et al.  (1977).  Chromosome aberrations and sister-chromatid
exchange in workers in chemical laboratories and  a rotoprinting factory and in
children of women laboratory workers.   Lancet.  2:322.

GiRNER-SMIDT, P., and FRIEDHICH, U.  (1978).   The mutagenic  effect  of benzene,
 toluene and xylene studied by the SCE  technique.  Mutat. Res.  58(2-3):313»

HOLMBERG,  P.C.   (1979).   Central-nervous-system defects  in children  born to
mothers exposed to organic solvents during pregnancy.  Lancet.  2:177-179.

HODAK,  A., and  UNGVARY,  G.   (1978).   Embryotoxic effects of  benzene  and  its
methyl derivatives:   Toluene and xylene.  Toxicology.  11:55.
                                     11-28

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KIRKHART, B.  (1980).  Micronucleus Test on Toluene.  Prepared  by  SRI  Intkrna-
ticnal,  Menlo Park,  CA,  urder Contract No. 68-02-2917 for the U.S. Environmental
Protection Agency, Research Triangle Park,  NC.

KOGA, K., and OHMIYA, Y.   (1978).   Potentiation  of toluene toxicity by hepatic
enzyme inhibition in mice.  .J.  Toxicol. Sci.   3(1) :25-29.

LIJINSKY, W., and GARCIA, H.  (1972).  Skin carcinogenesis tests of  hydrogenated
derivatives of anthracene and  other polynuclear  hydrocarbons.   Z.-  Krebstorseh.
Klin. Onkcl.  .77:226.  (Cited in U.S.  EPA,  1980).

LITTON  BIONETICS,  INC.   (1981).   Mutagenicity Evaluation  of  Toluene—Mouse
Dominant  Lethal  Assay.   Final  Report.   Submitted  to  the American  Petroleum
Institute, Washington, D.C. in January 1981.   LBI Project  No.  21111-05.  Licton
Bionetics, Inc., Kensington, MD.  15 pp.

LITTON  BIONETICS,  INC.    (1978a).   Mutagenicity  Evaluation of Toluene.   Final
Report.  Submitted to the American Petroleum Institute,  Washington, D.C.  in  Kay
1978.  LBI Project No. 20847.  Litton Bionetics,  Inc., Kensington,  MD.   150  pp.

LITTON  BIONETICS,  INC.   (1978b).  Teratology Study  in  Rats.   Toluene.   Final
Report.   Submitted  to  the American  Petroluem  Institute,  Washington,  D.C.  in
January  1978.  LBI Project No.  20698-1.  Litton Bionetics,  Inc.,  Kensington,  MD.
17 pp.

LYAPKALO, A.A.  (1973).   Genetic activity of benzene and toluene.  Gig. Tr. Prof.
Azbol.   VJ_: 2^-28.   (In  Russian with  Engligh summary;  evaluation  based on  an
English  translation  provided by the U.S. EPA.)

MAKI-PAAKKANEN, J. et al.  (1980).  Toluene exposed workers and chromosome aber-
rations.  J. Toxicol. Environ. Health.  j6:775.

MASON, M.M., GATE, C.C.  and BAKER, J.   (1970.  Toxicology and csrcirogenesis of
various  chemicals  used  in the preparations of vaccines.   Clinical Toxicology.
Hi 185-201.
                                     11-29

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MCLAUGHLIN, J. et al. (196*0.  Toxicity of fourteen volatile chemical as measured
by the chick  embryo method.   Am.  Ind.  H%£.  Assoc. J_.   25:282.

MORTELMANS, K.E., and RICCIO, E.S.  (1980).  In vitro Microbiological Gentoxicity
Assays of Toluene.   Prepared by SRI International, Menlo Park, CA, Under Contract
No. 68-02-29^7 for the U.S. Environmental Protection Agency, Research  Triangle
Park, NC.

NAWROT,  P.S., and  STAPLES, R.E.   (1979).   Embryo-fetal  toxicity  and  teratc-
genicity of benzene and toluene in the mouse.  Teratology.  J9:41A.   (abstract).

NCI  (NATIONAL CANCER  INSTITUTE).  (1979a).   Bioassay of 2,5-Dithiobiurea  for
Possible Carcinogenicity.  National Institute of Health, DHEW Publ. No.   (NIH)
79-1387.

NCI (NATIONAL CANCER INSTITUTE).   (1979b).  Bioassay of 3-Nitrc-p-acetophenetide
for Possible  Carcinogenicity. -National Institute of Health, DHEW Publ.  No.  (NIH)
79-1388.

NESTMANN, E.R.;  LEE,  G.G.-H*;  MATULA, T.I.;  DOUGLAS,  G.R.;  and MUELLER, J.C.
(1980).  Mutagenicity  of  constituents  identified in pulp and paper mill  effluent
using the Salmonella/mammalian-microsotne assay.  Mutat.  Res .  79:203-212.

NTP  (NATIONAL TOXICOLOGY PROGRAM).   (1983).   Chemicals  on Standard  Protocol
(1/13/83). Bethesda,  MD:  NTP, Carcinogenesis Testing Program.   Available  from:
Technical Information Section,  Carcinogenesis  Testing  Program,  NTP, Lanaow.
Bldg., Rm. A306,  Bethesda,  MD 20205.

POEL, W.E.   (1963).   Skin  as a  test  site  for the  bioassay of  carcinogens  and
carcinogen- precursors.  Natl.  Cancer  Inst.  Monogr.   10:611.

POWERS, M.B;   (1979).  Memorandum for  the Record from the NTP Chemical  Selection
Group, Toxicology Branch, CGT, DCCP,   National Institute, Washington, D.C.,  May
25, 1979-
                                     14-30

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ROCHE, S,M.,  and MINE,  C.H.   (1968).   The  teratogenicity of some  industrial
chemicals.  Toxicol. Appl. Pharmacol.  12:327.

SNOW, L.; KACNAIR, P.; and CASTO, B.C.  (198O.   Mutagenesis testing  of  toluene
in Salmonella  strains TAlOO and  TA98,   Report  prepared for the  U.S.  EPA  by
Northrup Services, Inc., P.O. Box 12313, Research Triangle Park,  NC 27709.

TATRAI, E.; HUDAK, A.; and UNGVARY, G.   (1979).   Simultaneous effect on the  rat-
liver  of benzene, toluene, zylene and  CCL'J.   Acta. Physiol. Acaci.  Sci. Hung.
53(2):261.

TOUTANT,  C.,   and LIPPMANN, S.    (1979).   Fetal  solvents syndrome  (letter).
Lancet.   1(8130):1356.

.TRACOR JITCO,  INC.  (1982).   Report of Audit-IBT  Study  No. 8S62-08810, 2^-Month
.Chronic  Inhalation  Toxicity and Carcinogenicity Study  with Toluene  in Albino
Rats.  Prepared  by  Tracor Jitco,  Inc.,  Rockville MD,  for the  Chemical Industry
Institute of Toxicology, Resaarch Triangle Park,  NC.  January  15,  1982.

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           15.  SYNERGISKS AND ANTAGONISMS  AT THE  PHYSIOLOGICAL LEVEL

15.1.   BENZENE AND TOLUENE
     Arunai studies have  shown  that benzene and  toluene  may be  metabolized by
similar enzyme ays tens  in the  parer.chymal  cells  of  the  liver.    Pawar  et al.
(1976)  found  that   the  activities   of  hepatic  aninopyrine  N-detsethylase,
NADPK-linked  peroxidation,   and  ascorbate-induced   lipid   peroxidation  were
reduced,  while  acetar.il ide  hydrcxylast-  was  increased,  by  either  benzene
pretreata;ent  or  toluene pretreatment  in male  rats.    Induction  of aminopyrine
N-denethylase ar.d components of the electron transport system was seen when the
anisals were giver, p.ner.obarbital (Fawar  et  al.,  197c; Mungikar and Pawar,  1976).
When phencbarDital was coacxir.istered with  benzene or toluene, the  changes in the
activity of these enzymes produced  by  single  administration of the xenobiotios
were  attenuated  (Pawar  et  al., 1976).   That  induction  of   hepatic enzymes  by
phenobarbitai affects metabolism  of toluene  is indicated  by the  reduction of
toluene  toxicity  (decreasea narcosis)  in  f stale  rats or male  itice  given
phenobarbitai prior to intraperitoneal injection of toluene  (Ikeda and Ohtsuji,
1971;  Koga  ana   Ohaiya,   197c),  and   the  accelerates   excretion  of  toluene
metabolites froe  female  rats  as  described in Sections  12-3- and 12.U.  (Ikeda and
Ohtsuji, 1971).
     The following studies indicate that toluene has the  potential for altering
the fcioactivity  of  benzene  wr.en Riven in sufficiently large quantities.   When
benzene was given in combination with toluene, the conversion of benzene to its
metabolites (phenols) was suppressed  in rats  (Ikeda. et al.,  1972)  and  in  mice
(Andrews et al.,  1977;.  Ikeda  et al.   (1972)  administered a mixture of benzene
arid toluene  (equivalent to  110  oig  benzene/kg  and  iJ30 mg toluene/kg) intraperi-
toneal ly to female rats and  observed a reduced excretion  of  total phenols.  When
a mixture of  toluene  and  benzene  (110 mg toluene/kg  and  4*10 mg benzene/kg) was
administered, hippuric acid  excretion  was reduced  up  to k  hours after injection.
Induction of hepatic microsoraal  enzymes  by  phenobarbitai prior to administration
of the mixture alleviated the suppression.
     Andrews  et al.  (1977)  co-administered   ^40  or  880 mg/kg  benzene  and
1720 mg/kg toluene  intraperitoneally to  mice  and  found a significant  reduction
in urinary excretion of benzene metabolites and a compensatory increase of pul-
monary excretion of unmetabolized benzene.  When toluene  and benzene were coad-
                                      15-1

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ministered  by  subcutaneous injection, toluene did  not  significantly change the
total  amount of  benzene found in fat, liver,  spleen, blood, or bone marrow, but
it did reduce  significantly  the accumulation of metabolites  in  these tissues.
Coadrainistratior. of  toluene and  benzene also counteracted benzene-induced reduc-
                  cq
tion of red eel]  J'Fe  uptake in  developing  erythrocytes, suggesting  that the
myelotoxicity  of  benzene  might  be attenuated by toluene-inhibition of benzene
metabolism  in  the bone  marrow.    In  an  ir\ vitro study with  a  liver microsorae
preparation, Andrews and coworkers (1977)  determined that toluene is a competi-
tive Inhibitor of benzene metabolism.
     In the studies  of  Ikeda et  si. (1972)  and Andrews et  al.  (1977">,  benzene and
toluene were administered  intraperitoneally in large amounts.  Sato and Nakajima
(1979), however, used doses in  the range  of  2*4.2  to 390.6 mg/kg of benzene and
23.6 to U6G.8  rag/kg  of toluene tc assess the effects  of concentrations that Eight
be found in the  workplace.  They found that when benzene was given to rats in the
range of 24.2  to 97=7 mg/kg,  there was no  significant difference in the rate of
disappearance  of  benzene  from  the blood  whether  the  benzene was administered
singly or  in  coraDination  with  an  equimolar  amount  of toluene.   At  a dose of
390.6 mg/kg benzene, an equimolar dose of toluene  delayed the disappearance of
benzene from  blood,  and the excretion of  phenol was reduced.   A dose-dependent
inhibition of the metabolism of benzene  by  toluene was  found.   In a study of
human exposure,  inhalation of a mixture of  25  ppm benzene and  100  r?10  toiuene for
2 hours did not exert  any influence on  the  disappearance rate  of benzene and
tcluene in either blood or end-tidal  (alveolar) air when compared to inhalation
of either  solvent singly.  Desaturation curves  (concentration versus  time) for
blood or en^-tidal air for each solvent after  inhalation of  the specified mixture
were virtually identical  to desaturation curves obtained after inhalation of the
same  solvent  (25 ppm benzene  or 100 ppm  toluene)   by itself.    These results
indicate  that  in   the  range of  threshold  limit  value  "the   pharmacokinetic
processes ...  of absorption, distribution,  excretion, and metabolism  ef either
benzene or toluene  are not  influenced  by simultaneous  exposure to the other"
(Sato and Nakajima,  1979).  The  data for  the single-solvent  exposures had been
published previously (Sato et al., 1974);  details of the experiment  with toluene
were discussed in Section 12.4.
15.2.  XYLENES AND TOLUENE
     When 0.1  m£/kg  cr 0.2 mJL/kg toluene was co-administered  with similar  doses
of  m-xylene  intraperitoneally  into   male  rats,  the  amounts of  hippurjc and
                                      15-2

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o-methylhippuric  acid excreted  in urine over  a  period of  21 hours  were  not
different from the amount  of metabolites  formed  by single injections of  toluene
or m-xylene.   The velocity  of  excretion of  metabolites in  the  simultaneously
injected  group was  slightly slower  than that  in the singly  injected  groups.
Thus, simultaneous administration of the compounds does not significantly inter-
fere with the metabolism of  either compound  (Ogat.a and Fujii, 1979).
    To  study the excretion  kinetic interactions  between  toluene and  xylene,
Riihimaki (1979) determined the conjugation and urinary excretion of metabolites
of toluene  m-xylene,  benzoic acid and methylbenzoic  acid in vivo in one  adult
human  male.   Forty-one  mmol benzoic acid or  7-4 mmol methylbenzoic acid  was
ingested singly or in combination by the subject.  Urine was collected for 25 to
30 hours after ingestion;  the total  recovery  of  the ingested compounds with  the
exception of  one  sample  (84$ of  the dose excreted  in  that  case)  indicated that
all elimination  took place  via  the  kidneys.   Combined  intake  of methylbenzoic
acid and  benzoic  acid did  not significantly affect conjugation or excretion of
either'metabolite.   This study indicates that during simultaneous exposures to
toluene  and JB-xylene, even  at    relatively  high occupational  exposure  levels,
conjugation and excretion of metabolites are not  likely to be rate-limiting steps
except  under  conditions  of lirrxted availability  of glycine.
15.3-   TOLUENE AND OTHER SOLVENTS
     Simultaneous  intraperitoneal injection of  1.18  g/kg  toluene and 0.91 g/kg
n-hexane  into female  rats  did not affect the concentrations  of _n-hexane in  the
blood nor excretion  of hippuric  acid (Suzuki  et  al.,  197*O  .
     Impaired peripheral  (ta;l)  nerve function  (as indicated by'decreased motor
nerve conduction velocity  and  mixed  nerve  conduction velocity,  and increased
distal latency) was observed in rats after 8,  12, and 16 weeks'  exposure  to 1000
ppm ji-hexane  for 12  houru/day  (Takeuchi  et  al.,  1981).  Similar exposure to a
mixture  of 1000  ppm n-hexane plus  1000 ppm  toluene  resulted in only  slight
impairment, and exposure to  1000 ppm toluene alone had a negligible effect on the
above indices.  Clinical  signs  of neuropathy were not  observed  in any  of  the
groups  throughout the experiment.
     Coadministracion of  ethanol  by  ingestion  and  toluene  by inhalation
(1060 ppm,  6  hours daily,  5  days  a week for  *4 weeks)  to rats did not change  the
electrocardiogram, hematocrit values, or histological  and  histochemical struc-
ture of the heart.  Toluene  increased vascular  resistance of the myocardium  and
reduced  cerebral blood  flow, while alcohol   ingestion reduced  arterial  blood
                                     15-3

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pressure, the cardiac index, and blood flow to the myocardium, kidney,  skin, and
carcass.  Myocardial and cutaneous vascular resistance,  as  well as cerebral blood
flow, increased after alcohol ingestion.  li was concluded that combined exposure
to the  two substances  produced  additive effects on myocardial  vascular resis-
tance (Morvai and Ungvary,  1979).  During subchronic exposure of rats to toluene
and ethanol,  there  is  a potentiation of microsomal  and mitochondrial changes in
the liver (Hudak et al.,  1978).   Ethanol administered  to rats in  single  oral
doses of 4 g/kg enhanced the _in vitro  metabolism of  toluene without causing an
increase in  the microsomal protein and  cytochrome  P-<450 contents  (Sato et  al.,
1981).   The enhancement was greatest  (about twofold) at the time when ethanol was
disappearing  from  the  body,  i.e.,  16 to 18 hours after ethanol administration.
     Smyth et  al.  (1969)  suggested  in a  study  of  joint  toxic  action  that
perchloroethylene  is capable of enhancing  the toxicity of  orally  administered
toluene in rats.  Withey and Hall  (1975) observed that intubation administration
of trichloroethylene and toluene  to  rats,  in combinations  of mixtures  at  five
different dcse  levels,  revealed  a  departure from  an additive  model.   They
concluded that the  effect of co-administration  of the solvents  could  not  be
described in  terms  of synergism or potentiation until further studies were made.
     Ikeda (197^)  found that coadministration of trichloroethylene  and toluene
(730 mg/kg and J430 mg/kg,  respectively) to  rats by the  intraperitoneal route
reduced  the  amounts'  of  metabolites  of both  solvents compared  with  amounts
excreted after administration of either solvent alone.

15.4  REFERENCES

ANDREWS, L.S.;  LEE, E.W.;  WITHER,  C.M.; KOCSIS, J .J.;  and SNYDER,  R.    (1977).
Effects of toluene on  the metabolism,  disposition and  hemopoietic  toxicity of
(SH)benzene.   Biochem.  Pharmacol.   77(4):293-300.

HUDAK,  A.; SZEBERENYI,  S.; MOLNAR, J.; CSEH, I.; SUVEGES, M.; FOLLY,  G.; MANYAI,
S.; and UNGVARY, G.  (1978).  Effect on liver of  chronic exposure to toluene and
ethanol in rat. Acta  Physiol.  Acad. Sc i. Hung.   5U1-2): 128.

IKEDA,  M.  (1974).   Reciprocal  metabolic inhibition  of toluene and trichloro-
ethylene in vivo and in vitro.   Int. Arch.  Arbeitsmed.  33(2):125-130.
                                      15-1

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IKEDA, M.; OHTSUJI, H-;  and  IMAMURA,  T.   (1972).  In vivo supression of benzene
and styrene oxidation by coadjninisterred toluene in rats and effects of4 phenobar-
bital.   Xenobiotica.  2(2): 101-106.

IKEDA,  M.,  and OHTSUJI „ H.   (1971).  Phenobarbital-induced  protection against
tbxicity of  toluene  and  benzene  in  the  rat.    Toxicol.  A£p_l_.  Pharmacol .
20 rO: 30- '13.

KOGA,  K,,  and OHMIYA,  Y.  (1978).   Potentiation of toluene toxicity by hepatic
enzyme inhibition  in mice.   J_.  Toxicol.  Sci.  3( 1 ): 25-29.

MORVAI,  V., and UNGVARY, G.  (1979).  Effects of simultaneous alcohol and toluene
poisoning  on  the  cardiovascular  system of  rats.   Toxicol.  Appl.  Pharmacol .
50(3J.: 38 1-389.

MUNGIKAR,  A.M.,  and PAWAR,  S.S.   (1976).   The effect of toluene, phenObarbital
and 3-nethylcholanthrene on hepatic microsomal  lipid  peroxidation.   Curr.  Sci .
OGATA,  M., and FUJII,  T.   (1979).   Urinary excretion of hippuric  acid  and m-
methylhippuric acid  after  administration  rf  toluene  and  m-xylene mixture to
rats.  Int. Arch.  Occup.  Environ. Health.  ^3( 1): 1*5-51.

PAWAR,  S.S.;  MUNGIKAR,  A.M.;  and MAKHIJA,  S.J.   (1976).   Phenobarbii:al induced
effect  on  pulmonary and hepatic microsomal  ethylmorphine  N-dPScthylase and lipid
peroxidation   during  oral   intoxication of organic  solvents in  rats.    Bull .
Environ. Contam. Toxicol .   15(5) : 357-365.

RIIHIMAKI, V.. (1979).  Conjugation and  urinary  excretion  of toluene and m-xylene
metabolites  in a aan.  Scand. J_,  Work Environ.  Health.   5(2): 135-1*42.

SATO, A.; NAKAJIMA, T.;  FUJIWARA,  Y.; and HIROSAWA ,  K.   (1974).  Pharmacokinetics
of benzene and toluene.  Int. Arch. Arbeitsmed.  33(3) : 169-182.
                                      15-5

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   \
SATO,  A., and NAKAJIMA, T.  (1979).  Dose-dependent metabolic interaction between
benzene  and   toluene  ^n  vivo  and   ^n  vitro.    Toxicol.   Appj..  Pharmacol.
US(2):2^9-256.

SATO,  A., NAKAJIMA, T.  and KOYAMA, Y.  1981.  Dose-related effects of single dose
of  ethanol on  the metabolism  in rat  liver of  some  aromatic and  chlorinated
hydrocarbons.   Toxicol. Appl.  Pharmacol;  60: 8-15.

SMYTH, H.F., Jr.;  WEIL,  C.S.; WEST, J.S.; and CARPENTER, C.P.   (1969).  Explora-
tion of joint  toxic -action:  Twenty-seven industrial chemicals intubated in rats
in all possible pairs.   Toxicol. Appl. Pharmacol.  11(2):340-3^7.

SUZUKI, T,; SHIMBO,  S.; and NISHITANI, H.   (WO.  Muscular atrophy due to glue
sniffing.  Int. Arch. Arbeitsmed.  33(2):115-123.

TAKEUCHI, Y.;  ONON,  Y.; and HISANAGA, N.  (1981).  An experimental study on the
      I
combined effects  of  n-hexane  and toluene on  the peripheral  nerve of  the. rat.
Brit. Jour  Indus. Med.  _3_8: 14-19.

WITHEY, R.J.,  and HALL,  J.W.   (1975).  Joint toxic  action of perchloroethylene
with  benzene or toluene in rats.  Toxicology.  M(1):5-15.
                                      15-6

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                         16.  ECOSYSTEM CONSIDERATIONS

16.1.  EFFECTS ON VEGETATION
16.1.1.  Introduction.  Toluene volatilizes  rapidly from solutions (Mackay and
Wolkoff, 1973)«   Most  studies investigating the phototoxicity  of  toluene have
been with  algae.   Of  thsse  studie's,  only one (Dunstan  et al.,  1975)  was done
under conditions  that maintained a nearly constant  concentration of toluene in
the  culture  medium throughout  the  experiment.   Other studies  were  done with
culture vessels capped  with metal caps or with cotton plugs, allowing the toluene
to volatilize and escape from the exposure solutions.  Even though steady-state
concentrations  are  lacking,  these  studies  do approximate  situations in  the
environment where a point source of  toluene  exists  to  a body  of  water.   The
discussion of these studies will, therefore, be  under the headings of "closed"
and "open" experimental systems.
16.1.2.  Effects of Toluene on Plants.
     16.1.2.1.  ALGAE
     16.1.2.1.1.    Closed  .System Studies — Dunstan  et  al.  (1975)  exposed  *4
marine  algal  species  to toluene  concentrations  ranging  from  1  to  10
Axenic algal cultures were inoculated at  18=C and grown with a  12-hour light/dark
cycle  under  cool-white   fluorescent  light  C4000 nW/cm ,   380   to  700 nm)  in
filtered enriched  seawater.   To minimize loss  of  toluene  by vaporization,  thj
125 m£, Erlenmeyer  flasks  were made airtight  with  rubber stoppers.   Experiments
were never run beyond a cell  density at which C0? limitations might limit growth.
The four species used were the diatom, Skeletonema costatum; the dinoflagellate,
Amphidinium carterae; the cocolithophorid, Cricosphaera carterae; and the green
flagellate, Dunaliella tertiolecta.
     To  illustrate the difficulty  of establishing  absolute concentration when
working with toluene, Dunstan et al. (1975) observed the toluene concentrations
at three intervals in stoppered flasks (Table 16-1).  Eighty-four percent of the
theoretical initial  concentration  was lost at  the  beginning  of the experiment
during the  handling and dispensing of the toluene into culture flasks, even when
the toluene was rapidly dispensed  under sterile conditions.
     Figure 16-1 shows how toluene can  both  stimulate and inhibit algal growth
depending on the species  and the concentration of toluene.  The dinoflagellate,
Amohidinium carterae. was inhibited at all concentrations of toluene (1 to
                                      16-1

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                                 TABLE 16-1

               Concentrations of Toluene in Stoppered FJasks
       Time of Measurement                         Percent  of Theoretical
                                                       Concentration


       Theoretical  initial  concentration                1C?

       Measured initial  concentration                    16

       Concentration after  3 days  of growth

          Stoppered flask                                14

          Cotton-plugged flask                            1

Source:  Dunstan et al., 1975
                                       16-2

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    150-,
UJ Z
UJ
K
I

O
cc
o
  UJ
  a.
    100-
     50-
0-fv\A
  1
     150-,
                                                    150-,
                 Amphidmium cartorM
                      UJ Z

                      P °
                         3£
                         Q.
                      UJ 	

                     ' I O
                                                   100-
                                                    50-
                10
103
                                 10*
10
                CONCENTRATION
Ul Z
-> o.
UJ —
CC -I
l§

§1
    100-
     50-
                 Skeleton»ma costatum
                102      103      104
                CONCENTRATION (jif /8)
                      uj z

                      ^ ec 100-
                      -J Q.
                      UJ —
                      EC _l
     §5  50
     c

                                                 °
                                          105
                                       Dunsliefla tertk>l»cU
^Qi      'O3
CONCENTRATION
10*
10a
                                      Cricotphaoru cwtaro
                     10Z      10J
                    CONCENTRATION
                                                       TO4
                                  FIGURE 16-1

  Phytoplankton Growth in  Various Concentrations  of Toluene (Organisms  were grown
  in  stoppered flasks.  Growth,  cieasured by cell  numbers and in vivo  chlorophyll,
  was determined on the 2nd  and  3rd days of logarithmic growth.  Concentrations of
  low molecular weight hydrocarbons are In theoretical values.)

                         Source:  Dunstan et  al., 1975
                                         16-3

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10  ng/X.)  from 20 to 50%.  The other  three species, however, were stimulated by 1
      li
to  10  ng/H,  but-   higher  concentrations  of   toiuene  either  had  no  effect
(Dunaliella  tertiolecta)  or  became  inhibitory   (Skeletonema   costatum  and
Cricosphaera carterae).  This  work indicated that  one  of the most significant
environmental effects was in the short-term selection of certain phytoplanktonic
species  by  the  growth  stimulation  brought  about  by  low levels  of  toluene.
Dunstan et al. (1975) concluded that the differential growth of phytoplanktonic
species within the phytoplankton population ultimately determines the community
structure, its succession,  and its trophic relationship.
     Potera  (1975)  evaluated  the  effect of  toluene on  saltwater phytoplankton
dominated by  Chlorella  sp.  using  Warburg manometry.  Toluene inhibited photo-
synthesis 29% at 3^ mg/4, and 35$  at  342  mg/2. (at 20=C).  Respiration (at 20=C)
wan inhibited 62% at 34 mg/JL and 16J at 342 mg/fc.
     16.1.2.1.?.  Open Studies —  Illustrative of the "open" type of experiment
is that of Kauss and Hutchinson (1975).  The freshwater alga, Chlorella vulgaris,
was exposed to toluene  for  10  days in  125 m2, cotton-plugged Erlenmeyer flasks.
Each flask was agitated to resuspend  the  cells daily.  The concentrations listed
in Figure  16-2  are nominal initial  concentrations.  In  this open experiment,
toluene was less toxic to the alga because the toluene concentration diminished
by volatilization  during the  experiments.   Comparison with  controls  revealed
that  a  lag  phase  that lasted  for  1  day  existed between inoculation  and
commencement  of  growth  for  50 and  100 mg/J,.   Recover;,  was less  rapid  with
250 mg/i.  At  concentrations  approaching toluene  saturation  (i.e., 505 mg/£),
toluene was lethal to the cells.
     Table 16-2 summarizes the toxic effects of toluene on algae.  In assessing
the toxicity of toluene  to algae,   both the inherent toxicity of toluene and the
exposure time need to be considered.  The no-effect  concentration  for sost algal
species studied appears  to be  at  the  1C  mg/£, level.  The evaporation rate from
solution (fresh or saltwater),  however,   rapidly  diminishes the exposure concen-
tration  of  toluene (Dunstan et al . ,  1975).   The  toxicity of toiuene  is more
closely approximated by  levels of  100 mg/i. in "open" systems, as  ?hown by Kauss
and Hutchinson (1975).
     16.1.2.2.   EFFECTS ON  HIGHER  PLANTS  — Currier  (1951)  exposed  barley,
tomatoes, and carrots to toluene vapor.   Air  at a flow rate of 11.5 K./min passed
through a small vaporizing chamber containing the toluene  and into  the top of a
bell jar containing the  plants.  The concentration  of toluene in  ~he vapor
                                      16-4

-------
  3
  CM
             .  O——O lOOppm
                A    • * ZSOppro
                               468
                                   T!ME(DAYS)
                               FIGURE 16-2

Growth of Chlorella vulgaris in Medium Containing Toluene  (Data plotted are
the average of three replicates.  Lines of best  fit  were determined using
regression coefficients.   Numbers represent initial  hydrocarbon concentration
on a parts per million basis.   The arrow on the  ordinate indicates starting
cell concentration.)

                       Source:   Kauss and Hutchinson,  1975
                                    16-5

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                                    TABLE 16-2

                        Toxic Effects of Toluene to Algae
Species
Concentration
   Effect
Reference
                                    FRESHWATER
Chi or el la vulgaris       245 mg/J.


Chlorella vulgaris       250 mg/X.


Microcystis aeruginosa   105 mg/i,


Scenedesiaus quadricauda  >400 mg/£
                 2H h ECCO
                 (cell number)

                 96 h no-effect  cone.
                 (cell number)

                 8 d no-effect cone.
                 (chlorophyll _a)

                 8 d no-effect cone.
                   (chlorophyll  a)
                       Kauss and Hutchinson,
                          1975

                       Kauss and Hutchinson,
                          1975

                       Bringmann and Kuhn,
                          1978

                       Bringniann and Kuhn,
                          1978
                                    SALTWATER
Amphidinium carterae     <0.001 Eg/2,
Dunaliella tertiolecta   10 mg/i
Skeletonena costatura
Ectocarpus sp.


Enteromorpha sp.
  10 mg/S,
Cricosphaera carterae    10 mg/i.
   1730 mg/S.


   1730 mg/«.
2 to 3 d  no-effect
cone, (cell number
and chlorophyll)

2 to 3 d  no-effect
cone, (cell nuinber
and chlorophyll)

2 to 3 d no-effect
cone, (cell number
and chlorophyll)

2 to 3 d no-effect
cone  (cell nuinber
and chlorophyll)

inhibits asexual
spore germination

inhibits asexual
spore germination
                                        Duns tan  et  al.,
                                           1975
                                        Duns tan  et  al.,
                                           1975
Dunstan et al.,
  1975
                                        Duns tan  et  al.,
                                           1975
Skinner, 1972


Skinner, 1972
h s hour; cone. = concentration; d = day.
                                       16-6

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chamber was varied by changingithe temperature of the toluene.  The concentration
of vapor in the air  was determined by measuring  the amount of toluene evaporated
per unit  of  time.   Three  tomatoes,  20 carrots,  and  12 barley  seedlings were
tested 32, 32, and T< days respectively after planting.  Plants were exposed in
the gas chamber  for 1M, 1/2, 1, and 2 hours.  Ths type  and extent of injury were
recorded after 'i month  to allow for a recovery period.  Temperature  of the plants
was held at 25°C.
     Results showed that toxic effects  of  toluene vapor were influenced by expo-
sure  period  and dosage  (Table 16-3).   Toluene  was  observed  to  be  toxic  at
concentrations of 6.4  to 12.0 mg/JL after  15  minutes of  exposure  (Currier,  1951).
Fifteen minutes of exposure at  12 mg/i. toluene produced a 50.  0, a:>J 60J injury
to  tomato, carrot,  and barley, respectively.   The  effects  of ^he exposures on
flower and fruit  development were not determined.   For lethality to  occur at
12.0 mg/i, barley required 1 hour, tomato 2  hours, and  carrot over-  2 hours.  The
toxicity  appeared  to  vary  markedly  within  a narrow  limit.   By  lowering  the
concentration of  toluene from 12.0  to 6.4 mg/S,, the  percentage  of  injury to
barley  after  a 2-hour  exposure was reduced from  100$ (lethal)  to  15J.   At
24.1 mg/£, toluene was  only  twice as toxic  to barley seedlings as at 12.0 mg/S,
after a 30-minute exposure.
     Toluene entered the plant rapidly  through the cuticle and  stomata.  Symptoms
of injury included a darkening of  the tips of the  youngest leaves,  presumably as
a  result  of  leakage  of sap  into the cellular  spaces (Currier,  1951).   This
darkening spread to the older  leaves.  There was  a loss of turgor, with draping
stems and leaves.  In bright sunlight, the  chlorophyll  was destroyed.
     Toluene  is  classified as  a  contact  poison  that  quickly  kills  the plant
tissue  with  which it  comes  in contact (Currier, 1951).  This  material  is not
accumulated in plants nor is it translocated.  The nech.anj.sm of toxicity involves
disorganization of the  outer membrane  cf  the cell due  to solvent  action on the
lipoid constituents,  resulting in  disruption of  photosynthesis,  respiration, and
turgor pressure.
16.2.  B10CONCENTRATION, BIOACCUMULATION, AND BIOMAGNIFICATION  POTENTIAL
     Limited information is available concerning toluene's potential for accumu-
lating  in aquatic  organisms  and  aquatic  food  chains.  Possible pathways of
toluene uptake *"••? dirtily  rrom water  (bioconcentratlon) and from both water and
food  (bioaccumulation).  Biomagnification occurs  if the concentration of a com-
                                      16-7

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                                     TABLE 16-3
          Toxic Effects  of Toluene Vapor pn Carrots,  Tomatoes, .and Barley
Percent Injury
Material
Tomato
Carrot
Barley
Barley
Barley
Concentration
12.0 mg/JZ.
12.0 ffig/ I
12.0 mg/i
6.H mg/£.
2U.1 mg/J.
Exposure Time (h)
1/4
50
0
60
0
ND
1/2
60
50
50
25
100
1
75
75
98
15
100
2
100
75
100
15
ND
 Source:  Currier,  1951
 0? =  no effect;  100} = lethal  1 month after exposure.
h = hour; ND = not  determined.
                                       16-8

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pound in an  organism increases with its  trophic  level  as a  result  of passage
through rood chains.
     Nunes and Benville (1979) studied the uptake and depuration of toluene and
other monocyclic  aromatic components  of  the  water-soluble  fraction  (WSF)  of
Alaskan Cook Inlet crude  oil  in  Manila clams (Tapes aemidecussata).  Claina were
exposed for 8 days to a constant WSF concentration under continuous-flow exposure
conditions.  The toluene concentration in  water was  measured daily.  The toluene
concentration in a pooled  sample of 10 clams was measured at 2, 4, 6, and 8 days.
At  the  end  of the exposure  period,  remaining  clams were  transferred  to
clean-flowing seawater and pooled tissue samples were analyzed for toluene after
1, 7, and 12 days of depuration.   The data are provided  in the following tabula-
tion:

                                        Toluene Concentration (ppm)
Exposure
Depuration
           Davs
1
2
3
14
5
6
7
8

1
7
                        Water
                                      1.2
                                      1.3
                                      1.7
                                      1.14
                                      1.2
                                      0.9
                                      1.0
                                      1.1
Tissue


 2.3
 2.2
 0.87
 2.0
                                                                 3.30
                                                                 0.80
                                                                 1.10
The  mean water  concentration during  the uptake  period was  1.2  ppm  toluene.
Tissue  concentrations reached  a maximum  by 2  days  of exposure  and  remained
relatively  constant  except for  a  temporary decline  on day six.   The average
tissue  concentration  during the exposure  period was  1.5 ppm.   The calculated
bioconcentration factor (BCF) is 1.25  (which is  equivalent to 1.5 ppm in tissue
and  1.2 ppm in water). The depuration study showed that to_luene  was  lost rapidly
during  the  first  week of  depuration,  but that  a  significant  concentration of
toluene remained in the clams by 2 weeks after beginning depuration.
                                                                    iij
     Hansen et al. (1978)  investigated the uptake and depuration of   C-toluene
by blue mussels  (Mytilus  edulis).   Groups of mussels were exposed  under static
conditions to four concentrations of   C-toluene for up to 8 hours, followed by
                                      16-9

-------
exposure to clean, recirculating seawater for up to 192 hours.  The   C-toluene
concentration in water and tissue (pooled sample from four mussels) was measured
by liquid scintillation  counting at 1,  2,  *»,  and 8 hours  after  beginning the
uptake phase and periodically in tissue  during the depuration phase.
     The   C-toluene concentration in tissue exceeded the water concentration by
1 hour at all exposure concentrations except the highest (^0 pi/kg = ppm), which
was toxic  as shown  by closure  of  the  missels at this concentration  (Hansen
et al . ,  1978).  Equilibrium was  reached by 1 hours  in all groups.  The BCF values
at 8  hours,  expressed as  the  tissue  concentration divided by the mean  water
concentration, were as follows:
                      Water concentration
                                                 BCF
                              0.05               3-8
                              O.H                5.7
                              4.0                3.6
                              U.O                3.6

     The BCF values,  which averaged U.2, seemed to be  independent of the exposure
concentration,  indicating  that accumulation  was proportional to  the  level  in
water (Hansen et al., 1978).  More than half of  the accumulated   C-toluene was
eliminated  by  1  hour after the depuration  phase began at all exposure concen-
                                            i ii
trations.  The depuration time by which no   C-tbluene was detectable in tissue
                                              1 4
was 1 hour  in  the  mussels  exposed to 0.05 ui   C-toluene/kg, 4 hours for those
exposed to O.U uA/kg, 120 hours for those exposed to 4 jiJl/kg, and 192 hours for
the animals exposed to UO ufc/kg.
     Lee et al . (1972) reported that the same  species of mussel (Mytilus edulis)
took up 3 to 10 ng  of  C-toluene  per mussel (average dry weight tissue r 0.3 g)
during static  exposure  for an  unspecified  period of  time  to 0.1  to 0.5 mg/Jl.
Using tissue toluene concentrations of  10 to  33  ug/g,  the BCF is calculated to
have been, between  66 and  100.   Because  these  values  are  based  on  dry tissue
weights rather than wet weight,  they are considerably higher  than those reported
by Nunes and Benville (1979) and Hansen et al. (1978).
     Berry  (1980)  investigated  the uptake  of l4C-toluene  by bluegill sunfish
(Lepomis macrochirus) and  crayfish (Orconectes  rustlcus).   The  exposure solu-
                                      1ii
tions were prepared by adding  1  mJl of    C-toluene to  100 I of  water for the fish
                                     16-10

-------
 experiment and by  adding 1  mS,   C-toluene  to  10 I of  water  for  the  crayfish
 experiment.   A group of 1*0 animals was added after thorough mixing of the solu-
 tions.  Duplicate water samples and 2 to 4  animals were taken at 0, 0.5, 1, 2,  4,
 8,  12,  16,  20, 2^,  and  48 hours  after beginning  exposure.   The    C-toluene
 concentration, expressed as  nanograms per milligram ( =  ppm),  was  determined  in
 water and in 7 (crayfish) or 9 (fish) tissues or organs  by liquid  scintillation
 counting.  The  BCF for  each  tissue  was  also  calculated.   Analysis  of  water
 samples  showed  that the  toluene concentration  in water  decreased  at  a  much
 greater rate  in the  crayfish  experiment than  in the  bluegill experiment  (89J
 versus 51? loss by 48 hours).  The  maximum  BCF of bluegill tissues ranged from
 about 3 for  brain  to 45 for  spleen.  Fish muscle  tissue was not analyzed.   The
 maximum BCF for most  fish tissues was reached by 8 hours.  The maximum  BCF  of
 crayfish tissues ranged from about 8 for muscle to 140 for hepatopancreas.  The
 B.CF values  increased  throughout  the 48-hour  exposure  period for all  tissues
 except testes and muscle.   These results indicate that toluene is accumulated
 above the water concentration by many tissues in these two species.  The BCF  of
 eight in the edible portion (muscle) of crayfish  is considered to be a minimum
' value  bf .ause  of  the  rapidly decreasing toluene  exposure concentration  during
 this experiment.
      Berry et al.  (1978) also measured the uptake of ^K-toluene by fed and unfed
 mosquito (Aedes aegypti)  larvae and the uptake of   H-toluene by fed larvae in the
 presence or  absence of  benzene.  The  larvae were exposed  to an initial concentra-
 tion of 0.5 m£ ^H-toluene/Jt, water.   Duplicate  water samples and  2 to  5  larvae
 were taken at  1, 2, 4,  8,  12,  15,  20, and 24 hours and  counted individually  by
 liquid scintillation counting.  Maximum ^H-toluene counts  per  minute (cpm) were
 equal  in  fed  and unfed  larvae,  but were reached more  quickly (1  hour  versus
 4 hours) by  the fed animals.  The  K-toluene counts per minute  values in larvae,
 expressed as  the  percentage of  initial  water  counts, were greater  during the
 first 4 hours  in the benzene and toluene mixture than in  the solution containing
 toluene alone.  BCF values  cannot  be calculated  because the authors expresssed
  H-toluene uptake as counts  per minute per larvae rather than  counts per minute
 per gram.  The weight  of the  larvae was not provided.  Interpretation was also
 complicated  by rapid loss of ^H-tbluene (half-time about four  hours)  during the
 uptake  period.   It  is  likely,  however,  that uptake by  ingestion  of toluene
 adsorbed to  food particles can  be a significant route of accumulation in aquatic
 organisms.
                                      16-11

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     Ogata and Miyake (1973)  identified  toluene as the cause of offensive odor in
the flesh of grey mullet (Hugil Japanicus) taken from a harbor receiving efflu-
ents from  refineries  and  petrochemical  industries.  Toluene  was identified in
seawater and  fish  tissue  by  gas chromatography, infrared  (IR) and ultraviolet
(UV) absorption, cind mass spectrotnetry.   The toluene concentration in most fish
was not quantified; however,  the flesh of  one mullet with an offensive odor con-
tained 5 pp«  toluene.   Additional  exparioNentj showed  that  toluene was accumu-
lated by caged  eels  kept  for 10 days in several locations  in  the  harbor  to an
average of 2.** times the water concentration-.  These eels had  the same offensive
odor as mullet collected from the harbor.  In another experiment, four eels were
exposed in seawater  to  which a mixed solution of benzene,  toluene, and xylenes
was added daily for 5 days. The concentration  of  each chemical  was then measured
in seawater, muscle, and liver.  The results with toluene were as follows:

                                Toluene Concentration
                  Fish No.      	(ppm)  	     BCF
     Muscle           1                 11.2              0.70
                      2                  2.6              0.16
                      3                  5.1              0.32
                      U                 30.8              1.91
                   Mean                 12.4              0.77
     Liver            1                  9.0              0.56
                      2                  2.5              0.1C
                      3                  5.2              0.32
                      U                  2.5              0.16
                   Mean                  U.8              0.30
     Water           —                 16.1

The results indicate that BCF  in muscle was equal to or greater than the BCF in
liver and that tissue concentrations rarely exceeded the water concentration.
     In later experiments,  Ogata and Miyake  (1978)  found  that  eels (Anguilla
Japonica)  accumulated toluene  to  whole-body  concentrations  greater  than the
water  concentration  in  freshwater.   For this study,  the  authors  studied the
uptake and  elimination  of toluene by eels  exposed  in  freshwater to crude oil.
The animals were exposed for  10 days to a recirculating oil suspension (50 ppm,
w/v), yhich was renewed every day.  During this period,  the toluene concentration
was  meadured  in  pooled groups  of  5 eels  taken on  1,  5,  and  10  days  after
beginning exposure.  The concentration of toluene in water was measured each day
                                     16-12

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at 1,  3, 6, 9, 1J4.5, and 24 hour? after preparing the crude  oil suspensions.  The
regaining eels were  then  transferred  to clean seawater and sampled after 3> 5,
and 10 days of depuration.  The average toluene concentration in Water during the
uptake period was 0.130 ppm.   The concentration in eels was  0..641  ppm after
1 day, 1.547  ppm after 5 days, and 1.718 ppm after  10 days.  The respective BCF
values were  4.9,  11.9, and  13.2.   A  semilogarithmic plot of  the logarithm of
tissue concentration versus  time indicated  that equilibrium  had not quite been
reached by  10 days.  The  depuration  phase of the experiment showed that tissue
concentration decreased rapidly from  1.718 ppra at the beginning of depuration to
0.31b ppm after 3 days, 0.121 ppra after 5 days, and  0.035 ppm after 10 days.  A
sem'ilog plot  showed  that  toluene was  eliminated  in  2 phases.   The elimination
half-time during the first phase, lasting from 0 to 5 days, was  1.4  days.  About
93J of the  accumulated  toluene  was  eliminated by the end  of this  period.   The
remaining toluene was eliminated at a somewhat slower rate, with  about 2% of the
accumulated toluene  remaining after 10 days of  depuration.
     The  only information  found  concerning fo.od-chain transfer  of toluene is
provided by Berry and Fisher (1979). who exposed mosquito larvae  (Aedes aegypti)
    • ji
to    C-toluene for  3 hours  and then  fed them  to  bluegill  sunr'ish  (Lepomis
macrochirus).  In duplicate  experiments, e,;ch of  25  fish in separate containers
were  fed  with 10  contaminated  larvae.   The mean level  of radioactivity in 10
larvae was 736 cpm in the first experiment and 3196 cpm in the second experiment.
Internal  organs  (spleen,  gall bladder,  liver,  stomach,  intestine,  and kidney)
from 5 fish, sampled  at  each  interval of 1, 4,  8, 24,  and 48 hours after feeding,
were analyzed for radioactivity by  liquid scintillation counting.  Radioactivity
was expressed as counts per minute  per organ rather than or. a weight basis.  The
only  organ  that  had  counts  per  minute  values significantly  greater than back-
ground levels was the stomach at 1, 4,  and 8 hours after feeding.  The authors
concluded that an insignificant amount  of toluene,  if any, leaves the digestive
tract  to  be  accumulated  in other  organs  of  sunfish.   The  validity  of  this
conclusion  is unknown because  the  dose was so  low  that absorption,  if it had
occurred, could not  have been differentiated from background counts and because
the counts were not  expressed on a tissue weight  ^asis, even in  the stomach.
     In summary,  the available information indicates that the  primary path of
toluene  uptake in  aquatic  organisms  is  direct  absorption  from  water.   The
reported or calculated BCF  values  for the edible portion or the whole organism
ranged from <1 to  about 14,  indicating that toluene  has a low bioconcentration
                                      16-13

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potential.   These BCF values are lower than the value predicted on the  basis of
the relationship established between octanol-water partition coefficient  (P) of
lipophilic  compounds and .steady-state BCF (Veith et al.,  1979).  This relation-
ship,  expressed by the equation I:log BCF 5 (0,85 log P)  -  0.70," would predict a
BCF of 39,  using a log P value of 2.69 for toluene (see Subsection 3.4.2.).
     Low bioconcentraton potential,  rapid depuration,  and  the ability of fish to
metabolize  toluene all indicate that  toluene  is unlikely to biomagnify through
aquatic food chains.  Aquatic organisms do accumulate  toluene,  however,  and con-
centrations in edible species from polluted  areas have reached  levels that cause
organoleptlc effects'in humans (Ogata and Miyake, 1973).
16.3.  EFFECTS ON MICROORGANISMS
     Toluene  has  been  used  for  quite  some  time  as  an  antimicrobial agent.
Sabalitschka and Preuss (1954) sterilized a urine sample containing Escherichia
coli  and  Pseudomonas  fluorescens   within   24  hours  with 4000 mg/S,   toluene.
Threshold concentrations for toluene have been established by Bringmann  and Kuhn
(1959.,  1976,  I960)  and Bringmann et al.   (1977)   for various microorganisms.
These investigators reported  values  of 29 mg/JL for P_.  putida, 200 mg/Jl for IE.
coli, and  greater  than 450 mg/2,  for  the ciliated protozoan  Uronema parduczi.
Partial sterilization of soil was  achieved by  adding toluene to the soil (Pochon
and Lajudie, 1948).
     The effects  of  toluene  on bacterial  activity and  growth  have also been
studied.  As measured by methane evolution rates, 20 mg/£,  toluene increased the
growth  rate  of bacteria in  sewage  sludge  deposits,  while 200 mg/S, produced a
toxic effect (Barash,  1957).  Similarly low levels  of  toluene allowed good growth
o' jp.  putida and Nocardia sp., while saturation levels (515 mg/& at 20°C) were
toxic (Gibson,  1975).   Depending  on the  concentration  (173  to 17,300 mg/X,), a
rotifer (Dicranophorus forcipatus) was unaffected,  or  temporarily inhibited, or
permanently inhibited by toluene (Erben,  1978).   Death  and  disintegration of
rumen ciliates occurred between 460 and 645 mg/2, of toluene (Eadie et  al.,  1956).
At  sublethal  concentrations  (1000  and 6000 mg/£,),  toluene  caused  a   negative
chemotactic response  or totally inhibited the chemotatic  response of all marine
bacteria tested (Mitchell et al.,  1972;  Ycung  and Mitchell, 1973).  Although the
effects were reversible, the authors of the  1972 paper expressed concern  that the
inhibition  could seriously undermine the capacity  of the marine microflora to
control  the  self-purification processes  in the  sea.  Beck  and Poschenrieder
(1963) found that high  concentrations of toluene  (50 to  100,000 mg/g of soil)
                                     16-14

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suppressed soil microflora activity.   In addition, they found that gram-positive
bacilli sporeformers, streptomycetes, and cocci were especially resistant, while
gram-negative bactrr-ia were sensitive.
     Toluene has been shown  to  affect the integrity of the inicrobial cell wall
and cytoplasmic membrane (Dean,  1978).  Thompson and  Macleod  (1974) reported that
marine pseudomonad cells washed and suspended in 0.5 M Nad  were lysed by treat-
ment with  20,000 mg/Ji  toluene  and  released 95?  of the cells'  alkaline phos-
phatase.   Because  the  cells remained  intact  with 0.05 M MgSO. and 20,000 mg/S.
toluene, the  authors concluded  that  Mg ions  prevented  cellular  disruption by
strengthening the integrity of the cell wall.   Woldringh  (1973) established that
a  2500 mg/2, solution  of  toluene partially  dissolved  the inner  cytoplasmic
memorane of E). coli and displaced nuclear material to the periphery of the cell.
DeSmet  et  al.  (1978) reported  that  at  100,000 mg/2, toluene,   the  cytoplasmic
membrane was completely disorganized.  The  presence  of Mg ions at lower toluene
concentrations  (up to  10,000 mg/2.),  however,  prevented extensive damage to the
cytoplasmic  membrane and  loss   of  intracellular material;  thus,  permeability
depended on the integrity  of the outer membrane (DeSmet et al.,  1978).  Deutscher
(197^)  found  that the  effects  of tolrene  treatment  were  dependent  on various
cultural conditions including pH, temperature, Mg ion concentration,  and age of
the  culture.    Temperature-dependent  effects  of  toluene   treatment  were  also
reported by Jackson  and  DeMoss (1965).   Toluene changed  the  asymmetric  unit
membrane profile to a symmetric profile in vegetative cells  of Bacillus subtil is
and  caused gaps  in  the  membrane to  appear  (Silva  et al.,   1978).    Fan  and
Gardner-Eckstrom  (1975)  found  that  toluene-treated  Bacillus  megaterium cells
liberated  a membrane protein essential  for-  peptidoglyca  synthesis and that this
protein could  be  added  back to the membrane to  reconstitute peptidoglycan syn-
thesis.    Toluene  at  86,000  mg/H   induced   the   autolysij  of  Saccharomyces
cerevisiae,  the release  of UV  absorbing  substances  from  the cells,  and  the
deacylation   of  phosphoplipids   (Ishida-Ichimasa,   1978).     At   saturation
concentrations  of  toluene,  however,  no cytolysis of yeast  occurred (Lindenberg
et al.,  1957).   Scholz et al.   (1959)  noted that  toluene-treated  yeast cells
accumulated hexosephosphates.   Bucksteeg (19^2)  found  that  the concentration of
toluene  and  time of  exposure  determined  its  effect  on  Cytophaga  sp.  and
time needed to produce  lethal effects.  Azotobacter was more resistant than the
Cytophaga  sp.   Bucksteeg  theorized that  toluene  affected  the  physical  and
                                      16-15

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i    chemic?! constitution of the cell.   An alteration in plaque  morphology in two
    coliphages (Tgrt and Tj occurred with 1% toluene (Brown, 1957).
         The ability of  toluene  to disrupt cell membranes  led  to the use  of this
    compound  as  an unmasking  agent in  microbial  research  to  assay a  variety  of
    enzymes (Herzenberg.. 1959;  Dobrogosz and DeMoss, 1963;  Levinthal et al., 1962).
    The _in vitro assays  using  toluene  have been used to make enzymes within a cell
    accessible to  exogenous substrates  (Jackson and DeMoss, 1965;  DeSmet  et al.,
    1978).  Generally,  toluene treatment makes the cells permeable to low molecular
 \  weight compounds (such as -deoxynucleoside triphosphate dNTP) and several macro-
    molecules  while remaining impermeable to  proteins  larger than ^pnroximately
    50,000 daltons  (Deutscher,  1974;  DeSmet et al., 1978).   Several investigators
    have  used these findings  to  study  DNA replication in  bacteria (E. c?li,  B.
    subtilis), bacteriophage (E. coli, Tj,),  and diatoms  (Cylindrotheca  fusiformis)
    after treating the organisms with 0.1 to 1? toluene in solution (Miller et al.,
    1973; McNicol and Miller,  1975; Moses  and  Richardson,  1970; Matsushita  et al.,
    1971; Winstun and Matsushita,  1975; Sullivan and Valcani, 1976).  Other uses  of
    toluene treated cells  are  in studying  the  synthesis  of heteroribonucleotides,
   ' RNA,  and  peptidoglycan and the  repair synthesis of DNA (DeSmet et  al.,  1978;
    Moses and  Richardson,  1970; Segev  et al.,  1973;  Winston and Matsushita, 1975).
    Burger (1971) showed that  toluene-treated  E_.  coli  cells continued DNA replica-
    tion,  but only in  that  chromosomal region  that  was  about  to  be  replicated'
    iri vitro.  Toluene-treated cells can also be used to study the  effects of various
    antibiotics in cell  growth and DNA  replication  (Hein,  195^; Burger  and  Glaser,
    1973).
         Although  the  exact mechanisms  of toluene-induced  disaggregation  of cell
    membranes are not known, Jackson and DeMoss  (19&5) state that the mechanisms fall
    into  two  classes:    (1) a disaggregating  (autolytic)  enzyme(s), perhaps  syn-
    thesized  in  the presence  of  toluene, or  (2)   a  direct denaturation  of  cell
    membrane   constituents   such  as  phospholipids;  a   condition  inhibited  by
    stabilizing factors such as divalsnt cations (e.g., Mg).
                                         16-16

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16.14.  REFERENCES

BARASH, V.A.   (1957).   The influence of some mineral and organic substances on
methane fermentation  in sewage sludges,   Vsesoyuz.  Nauch.-Issledovatel.  Inst.
Vodosnabshen.,  Kanalizats., Gidrotekh.    Sooruzhenii  i  Inzhener-  Gidrogeol,,
Materialy Soveschaniya.  pp. 105-114.

BECK, 1.  and POSCHENRIEDER, H.   (1963).   Experiments  concerning the action of
toluene en the microflora  in soils.  Platn. Soil.  18:  346-357.

BERRY,  W.O., BRAMMER,  J.D.,  and BEE,  D.E.   (1978).   Uptake  of water-soluble
gasoline  Tractions  and  the}'" effect,  on oxygen consumption in aquatic stages of
the mosquit^. (Aedes aegypti L.).  Environ. Pollut.   15(1):  1-22.

BERRY,  W.O.  and FISHER, J.W.   (1979).   Transfer of toluene  14C from mosquito
larvae  to bluegill  sunfish.  Bull. Environ. Contain. Toxicol.  23(6): 733-736.

BERRY, W.O,   (1980).  A  comparative  study  of  the uptake of  toluene by bluegill,
sunfish   Le-pomis  macrochirus   and  Orconectes  rusticus.    Environ.  Pollut.
80:  109-119.

BRINGMANN,  G.,  and  KUHN,   R.    (1978).   Grenzwerte der  Schadwirking wasserg-
efahrdender   stoff  gegen   blaualgen  (Microcystis  aeruginosa)  und  grunalgen
(Scenedestrus quadricauda)  in zelivermehrungschemmtest.  Vcm Wasser.  50: 45-50.

BRINGMANN, G., GOTTFRIED, and KUHN, R.  (1977).  Limiting values  for the damaging
action  of water  pollutants to  bacteria  (Pseudomonas  putida)  and  green  algae
(Scenedesmus quadricauda)  in the cell multiplication inhibition  test.  2. Wasser
Abwasser  Forsch.   10(3-4):  87-98.

BRINGMANN, G.  and KUHN,  R.  (1976).  Comparative results of  the  damaging effects
of  water  pollutants  against   bacteria  (Pseudomonas  putida)  and   blue  algae
(Microcystic aeruginosa).   Gas-Wasserfach, Wasser-Abwasser.   117(9): 41-113.
                                      16-17

-------
BRINGMANN, G. and KUHN,  R.  : (1959).  The toxic effects of waste water on aquatic
bacteria, algae, and small  crustaceans.  Gesundheis-Ingerieur.  80: 115.  (Cited
in McKee and Wolf, 1963),

BRINGMANN, G. and  KUHN,  R.  (1980).  Bestimmung  der biologischen schadwirkung
wassergefahodender atoffe gegen protozoen.  II.  Bnkterinpressende ciliaten.  Z.
Wasser Abwasser Forsch.  13(1); 26-31.

BROWN, A.   (1957).  Alterations  of plaque morphology in acme caliphages.   J_.
Baoteriol.  _73:  585-537.

BUCKSTEEG, W.  (1942).   The effect and mode of action of toluene en the bacterial
cell.  Zentr. Bakt. Parasitenk.  _105: 209-213.

BURGER, R.M.  (1971).  Toluene-treated Escherichia coli replicate only that DNA
which  was   about   to   be  replicated  _i£   vivo.     Proc.   Nat.  Acad.   Sci.
68(7): 2124-2126.

BURGER, R.M. and GLASER, D.A.  (1973).  Effect of nalidixic acid on DNA replica-
tion   by  toluene-treated   Escherichia   coli.      Proc.  Nat.   Acad.   Sci.
70(7): 1955-1953.

CURRIER,  H.B.    (1951).   Herbicida.i  properties  of benzene and  certain  methyl
derivatives.  Hilgardia.  20(19): 383-406.

DEAN,  B.J.    (1978).   Genetic  toxicology of benzene,  toluene, xylenes,  anc
phenols.  Mutat. Res.  _47: 75-97.

DE SMET, M.J., KINGMA,  J. and WITHOLT, B.   (1978).  The  effect of toluene on the
structure and permeability of the outer  ana cytoplasmic membranes  of Escherichia
coli.  Biochim.   Biophys. Acta.  506( 1): 64-80.

DEUTSCHER, M.P.    (1974).   Preparation of cells permeable to  macromolecules  by
treatment  with   toluene.    The  tRNA  nucleotidyltransferase.    J^  Bacteriol.
116(2): 633-639.
                                     16-18

-------
 DOBROGOSZ,  W.J.   and DeMOSS,  R.D.    (1963).    Induction  and  repression of
 L-Arabinose isomerase in Pediococous pentoaiaceuy.  J. Bacteriol.  85:  1350-1365.

 DUNSTAN, W.M., et al.  (1975).  Simulated and inhibition of phyto playlet on growth
 by iow molecular weight hydrocarbons.   Mar.  Biol.   31:  305-310.

 EADIE, J.M., MANN, S.O*. and OXFQRP, A.E.   (1956).   Survey of  physically active
 organic  infusoricidal cctnpounds  and  their  soluble derivatives  with  special
• reference  to  their action on the  rumen  microbial  system.  J.  Gen. Hicrobiol.
 _U: 122-133.

 ERBEN, R.   (1978).  Effects of some petrochemical  products on  the survival of
 Dicranophorus forcipatus 0. F. Muller  (Rotatoria)  under laboratory conditions.
 Verein. Linnol.  20:   1988-1991.

 FAN,  D.P.  and GARDNER-ECKSTROM,  H.L.    (1975).  Passage of a meiabrane protein
. through  the walls  of toluene-treated  Bacillus  megaterium  cells.   _J.  Bact.
' 123: 717-723.

 GIBSON, D.T.  (1975). Microbial Degradation of  Hydrocarbons.  E.D. Goldberg, Ed.
 The Nature of Seawater:  Report of  the Dahlem Workshop on the Nature of Seawater,
 Berlin,  1975, March  10-15.   Physical  and Chemical Sciences Research Report 1.
 Dahlem Konferenzen, Berlin,  p.  667-696.

 HANSEN, N., JENSEN, V.B.,  APPELQUIST,  J.,  and MORCH, E.  (1978).  The uptake and
 release of  petroleum  hydrocarbons  by  the  marine  mussel Mytilus edulis.   Prog.
 Water Technol.   10(5-6): 351-359.

 HEIN, H.   (195^^.  Bakteriologische Untersuchungen  mit  neomycin and nebacetin.
 Arznemittel-Forachung.  4_: 282-287.

 HERZENBERG, L.A.   (1959)-   Studies on the induction of beta-galactosidase in the
 cryptic  strain  of   Escherichia   coli.     Biochimica.  _et  Biophysica.   A eta.
 31: 525-539.
                                      16-19

-------
ISHIDA-ICHIMASA ,  M.   (1978).   Degradation of  lipids in  yeast  (Saccharomyce
cerevisiae) at the early phase  of  organic solvent-induced autolysis.   Agric.
Biol . Chem.  J2(2): 247-251.  Taken from:  Chem. Abst.  88;: I6482ilq,  1978.

JACKSON, H.W. and DeMOSS, J.A.  (1965).   Effects of toluene on Escherichia coli.
J_.  Bacteriol.  90(5):
KAUSS, P.B.  and  HUTCHINSON,  T.C.  (1975).   Effects  of water-soluble petroleum
components on the growth of Chlorella vulgaris.  Environ. Pollut.  9 ( 3 ) :  157-17^.

LEE, R.L., et al .  (1972).  Petroleum hydrocarbons:  Uptake and discharge by tLe
marine mussel Mytilus edulis.  Science.  177:
LEVINTHAL,  C.,  et  al .   (1962).   Reactivation  and  hybridization of  reduced
alkaline phosphatase.  Proc. Nat. Acad. Sci.  *I8_: 1230-1237.

LINDENBERG, B.A., MASSIN, M. and GAUCHAT,  G.  (1957).  Cytolysis of yeast caused
by narcotics considered at> an indifferent physical phenomenon.  Compt . Rend. Soc.
Biol.   151: 1369-1372.  Taken from: Chem. Abst.  52: 13856d, 1958.

MACKAY, D.  and  WOLKOFF , A.Q.   (1973).   Rate of  evaporation  of low-solubility
contaminants from water bodies  at  atmosphere.   Environ.  Sci .  Technol .   _7: 611.
(Cited  in Syracuse Research Corporation, 1980).

MATSUSHITA, T.,  WHITE,  K.P.  and SNECKA.   (1971).   Chromosome  replication  in
coluenized Bacillus subtilis cells.  Nature New Biol.   232: 111-1T4.   (Cited  in
Winston and Matsushita,  1975).

McNICHCL, A.L. and MILLER, R.C.  (1975).  Biological activity of Ty DNA  synthe-
sized in toluene treated Escherichia coli cells.  J. Virpl .  15: iJ79-'t83.

MILLER, R.C., TAYLOR, D.M.,  McKAY,  K,  and SMITH, H.W.  (1973).  Replication of Ty
DNA in  Escherichja coli  treated with toluene.  J. Virol.    12:  1195-1203.
                                     16-20

-------
MITCHELL, R., FOCEL, S. and CHET, I.  (1972).  Bacterial chemoreception.  Impor-
tant   ecological   phenomenon  inhibited   by   hydrocarbons.      Water   Res.
£(10): 1137-1140.  Taken from:  Chem. Abst.  7_8: 53571d, 1973.

MOSES, R.D. and RICHARDSON, C.C.  (1970).  Replication and repair of DNA in cells
of Escherichia coli treated with toluene.  Proc. Nat. Acad. Sci.  6j:  67^-681.

NUNES, P. and BENVILLE,  P.  (1979).   Uptake and depuration of  petroleum in the
manila  clam, Tapes semidecussath  Reeve.    Bull.   Environ.  Contain.   Toxicol.
21(6); 716-726.

OGATA, M.  and  MIYAKE,  Y.   (1978).   Disappearance  of aromatic  hydrocarbons and
organic sulfur compounds from fish flesh reared in crude oil suspension.  Water
Res.  12(12): 10M1-1044.

OGATA, M.  and  MIYAKE,  Y.   (1973).   Identification  of  substances  in  petroleum
causing objectionable odor in fish.   Water Res.  1_:  1^93-1504.

POTERA, G.T.   (1975).    The effects  of benzene, toluene,  and  ethyl benzene  on
several  important  members  of the  estuarine  ecosystem.    Piss. Abstr.  13.
36(5): 2010.

POCHON, J. and LAJUDIE, J.  (19^8).   Action of certain antiseptics on the normal
microflora of the soil.  Compt. Rend.  226: 2091-2092.

SABALITSCHKA, T. and PREUSS, J.  (1954).  Action of  toluene on  bacteria.  Deut.
Apqth.-Ztg. ver. Suddeut.  Apoth-Ztg,   £4: 1226-1228.

SCHOLZ, R.-, SCHMITZ, H., BUCHER, T. and LAMPEN, J.O.  (1959). Effect of nystatin
on yeast.  Biochem. Z.   331: 71-86.

SEGEV, N., MILLER, C.,  SHARON,  R. and BEN-ISHAI, R.   (1973). Exicision repair of
ultraviolet  radiaiton  damange in  toluene  treated  Escherichia  coli.   Biochem.
Biophys. Res. Commun.  J53CO: 12^2-1245.  Taken from:  Chem. Abst.   	: 62056n.
                                     16-21

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SKINNER, C.E.   (1972).   Role of algae  in  the  deterioration of decorative  and
marine paints.  FATIPEC Cong.  _H:  421-427.

SILVA, M.T., SUSA, J.C.F. and BLASSA,  G.   (1978).  Ultrastructural  effects  of
chemical agents and moist heat  on  Bacillus subtilis.   I.  Effects  on vegetative
cells.  Am. Microbiol.  129B: 363-375.  (Cited in NRC,  1980).

SULLIVAN, C.W. and VOLCANI,  B.E.  (19t6).   Role of silican in diatom metabolism.
VII. Silicic acid-stimulated DMA  synthesis in  toluene permeabilized cells  of
Cylindrothica lusiformis.  Exptl.  Cell Res.  98:  23-30.

THOMPSON, L.M. and McLEOD, R.A.  (1974).   Biochemical  localization  of  alkaline
phosphatase  in  the  cell  wall  of  a  marine  pseudomonad.    J.   Bacteriol.
117(2): 819-825.

VEITH, G.D., DEFOE, D.L., BERGSTEDT,  B.V.   (1979).  Measuring and Estimating the
Bioconcentration  Factor  of  Chemicals  in  Fish.   ^J.  Fish  Res.  Board  Can.
36: 1040-1048.

WINSTON, S. and MATSUSHITA,  T.   (1975).  Permanent loss of chromosome initiation
in toluene-treated Bacillus subtilis cells.  ^J. Bacteriol.  123: 921-927.

WOLDRINGH, C.L.  (1973).  Effects  of  toluene and  phenethyl alcohol  on the  ultra-
structure of Escherichia coli.   Jf-  Bacteriol.   114(3);  1359-1361.

YOUNG, L.Y. and MITCHELL, R.   (1973).  Negative chemotaxis of marine bacteria to
toxic chemicals.  Appl. Micro.   25(.6); 972-975.
                                     16-22

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                        17.  EFFECTS ON AQUATIC SPECIES

17.1.  GUIDELINES FOR EVALUATION
     Evaluation of the available  information  concerning the effects of toluene
on aquatic organisms  must  take into account  several  factors.   A  primary con-
sideration for evaluation of  toxicity test results is toluene's  high volatility.
The half-life for volatilization of toluene from a  water  column  1 m deep has been
reported to be between approximately 30 minutes (Mackay and Wolkoff,  1973) and
5 hours  (Mackay  and  Leinonen, 1975).   Benville  and  Korn  (1977)  analyzed the
toluene concentration in test  containers  during a 96-hour static toxicity test
and  showed  that  the  percentage  of toluene lost  was 48$  by 24 hours,  53%  by
48 hours, and greater than 99$ by 72 hours.   Korr. et al.  (1979) reported that
toluene was lost  at a greater rate  from bioassay containers at 12°C (99$ loss by
72 hours) than at 8°C (>99$ loss by 96 hours)  or at 4°C (75$ loss by 96 hours).
Potera (1975) found that the observed half-life of  toluene in bioassay containers
was  16.5 jf 1.13 hours.  The rate of volatilization of toluene from water varies
with the amount of mixing,  temperature, surface area to volume ratio,  and other
factors.  Adsorption to sediments and suspended particles may decrease evapora-
tive loss and result  in greater persistence of toluene.  Although adsorption may
lower  the  concentration  of  dissolved  toluene in the water column,  binding  to
sediment and suspended matter may increase the  effective exposure concentration
to benthic. and filter-feeding organisms.
     Most of the  reported  aquatic toxicity  studies  with toluene have  used  a
static exposure technique.    In most cases, the  LCCQ has been calculated on the
basis  of initial nominal (unmeasured)  or  initial  measured concentrations.  The
test organisms  in these static  experiments,   however,  are exposed to rapidly
decreasing toluene concentrations.  Most  of  the reported acute static toxicity
studies show little or no change in the LC5Q value  between  24 and 96 hours.  This
lack of change indicates  that most,  if not all,  of the mortalities in these tests
occurred during the first 24  hours when  toluene  concentrations were highest.  In
contrast, those flow-through studies that  reported acute LC™ values at more than
one exposure period showed that LCrQ values decreased significantly with time.
     Numerous  other   factors  may  affect  the  results  of  toxicity tests with
toluene.  It has been shown  that the  acute toxicity of toluene  is affected  in
some cases  by  temperature  and  salinity  (Section 17.3.).   These effects  on
                                      17-1

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toxiclty may be due to effects o'n the test organisms (metabolism, uptake, stress,
etc.), effects on the physicochemical  behavior  of  toluene  (solubility, volatili-
zation, etc.), or interactive effects of both.   For example,  toluene  is less
soluble in saltwater than in freshwater and is both more soluble and more vola-
tile at higher temperatures.  Laboratory results  may also bs influenced by t-he
loading ratio (gram  organism per liter water); dissolved oxygen concentration;
age, health, and  species of test organisms;  and  other  exposure conditions, all of
which may interact to affect the results in an unpredictable manner.
     Prediction of environmental effects from  laboratory  results  must consider
the  influence  of the variables  associated  with  laboratory  tests  and with the
natural variability  intrinsic .to the  aquatic  environment.   Results  of static
acute toxicity tests with volatile  compounds such  as  toluene may approximate the
acute toxic  effects  that  may  occar  in nature to  the  same species  during acci-
dental  spills,  because  toluene concentrations  rapidly  Decrease in both situa-
tions.   Flow-through acute toxicity  tests may provide some  insight  into the
expected effects of a short-term but constant release of toluene into the aquatic
environment,  as  might  occur  in  areas  receiving  refinery  or  petrochemical
effluents.  Neither static nor flow-through acute  toxicity tests can predict the
chronic effects  of  low  level toluene  pollution.   In  addition, acute toxicity
tests usually determine the concentration of toxicant that kills or affects 50%
of the test population.  LC™ or EC,-n values,  therefore,  represent concentrations
that  are toxic to half the population, and  provide no information concerning the
concentration that will  have no adverse effects during acute or  chronic exposure.
17.2.  EFFECTS OF ACCIDENTAL SPILLS'
     No  information  was  found  concerning  the  effects  of accidental  spills of
toluene per se on aquatic organisms; however, toluene is one of the major aroma-
tic components of crude oil and such  refined petroleum  products as  diesel fuel,
gasoline, anc1 jet fuel, all of which have been  released in large amounts to thn
aquatic environment  during spills.
     The long term ecological impact of accidental spills  of  toluene is unknown.
In spill situations, most of  the toluene would  probably evaporate rapidly.  For
instance, McAuliffe  (1976)  reported  that  toluene, benzene, and xylene could be
found in the water under crude oil slicks only  during the  first 30 minutes after
spillage.   In contrast, spills in areas  of shallow water and  restricted water
floW,  auch as in  certain portions of estuaries, lakes,  and  streams,  have  a
                                      17-2

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greater potential  for  causing acute mortalities because  the  toluene may reach
higher dissolved  concentrations and  may  persist longer  through  adsorption to
sediments.  Toluene is acutely  toxic  to many aquatic species  at concentrations
well below its water solubility, and lethal exposure may occur during spills in
shallow water.
     Although chronic,  low-level  pollution  by  toluene has been  reported  in a
Japanese river (Funasaka et  al., 1975) and a harbor  (Ogata  and Miyake, 1973) that
received  refinery  and  petrochemical  effluents,  the effects of such low level
chronic pollution in natural aquatic habitats are unknown.
17.3.  LABORATORY STUDIES OF TOXICITY
17-3.1.   Lethal Effects.  The lethal  effects of toluene have been reported for
numerous species of freshwater and marine fish and invertebrates.  The acute LCc-0
for  22 species  of  freshwater and marine  animals ranged between 3 ana 1130 ppm
(Table 17-1). All  but six of the LC5Q  values were determined in static tests.  Of
the  six flow-through LC   tests, four utilized measured toluene concentrations.
No information was found concerning the effects of toluene on amphibians.
     17.3.1.1.    FRESHWATER  FISH — The  earliest  investigation  of  toluene
toxicity to freshwater  fish  was conducted by Shelford et al.  (1917), who reported
that 1 hour  of  exposure to  61  to 65  mg/Ji toluene  was  lethal  to orange spotted
sunfish (Lepomis humilis).   This test was conducted under static  conditions at
20°C in freshwater of unspecified temperature and composition.
     Degani  (19^3)  conducted static toxicity tests  with  15-day-old  lake trout
(Salvelinus namaycush)  fry and 1.5 g mosquitofish (Gambusia  affinis)  in dechlor-
inated tapwater at  17 to 18°C using  3  to 5 fish per container (2 S, volume).  The
time  to  death   at a  nominal  exposure   concentration of  90 ppm toluene  was
390 minutes  for trout  and 47 minutes  for   mosquitofish.  The time  to  death of
trout fry exposed to 50 ppm  toluene was 258 minutes.
     Wallen  et  al.  (1957)  also conducted  static acute toluene toxicity tests
with female  mosquitofish  (Gambusia  affinis)  of  unspecified  size in turbid pond
water  (150 ppm  turbidity as  measured  by  Jackson turbidimeter, pH 7.5  to 8.5,
methyl orange alkalinity <  100 ppm,  temperature  17  to 22°C).  For these toxicity
tests,  ten fish per concentration  were  added   immediately  after addition  of
different amounts of toluene  to the bioassay containers (15 liter volume).  The
test solutions were constantly  aerated and mortalities were recorded daily for
96 hours.  The  24, 18, and  96  hour LC™  values were 13HO,  1260,  and "180 ppm,
respectively.  These values  were estimated  on the  basis of the initial nominal
                                      17-3

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                                                                        TABLE 17-1

                                                Acute Toielclty  of Toluene  to  fish  and  Aquatic  Invertebrates
Species
FISH
Freshwater
Ide

(Leuciscus Idus
aelanotus)



Hosqultof lah
(Gaobusla afflnls)

Goldfish
(Carasslus auratus)

Goldfish
— • (Carasslus auratus)
-j
Jr
Goldfish
(Carasslua auratus)









Fathead ralnncv
( Pioephales proeaelas)
LC50
Terap. Type 21 h 18 h 72 h
( °C) Test


20*1 SU 	 70

2Cj-1 SU — - 1?2




17 to SU 13110 1260 	
22

20^1 SM 56 	


25 SU 57.7 57.7
(18.9 (18-9
to to
68.8) 68.6)
17 to FM 11.6 J7.6 25.3
19 (32.0 (21.6 (20.1
to to to
71-7) 36.0) 3V. 9)







25 FM — 	

No Effect Reported
96 h Concentration Concentration Comments
Units


52 mg/t Lab 1, icoj kill at
BB.ng/l.
365 Lab 2, 1001 kill at
170 n«/L.
Teats were supposedly
conducted under
Identical conditions.
1180 560 ppn Tests were conducted
in afrated turbid
pond water.
	 	 Bg/t Test was conducted
in tap water ( pH
7.6)
57.7 — - ng/l Test was conducted
(18.9 in soft water.
to
68.8)
22.80 — - ppa Teats were conducted
(17.1 under flow-through
to to conditions in soft
30.0) dechlorinated tap
water. The test was
continued to 720 h
( 30 d) at which
time the LC (and
951 confidence Intar-
val) was ID. 6 ( 10.7
to 20.0) ppa.
18-7? 	 ng/l Eobryas were »ore
resistant than lartae
Reference


Juhnke and
Ludenann, 197B





Wall en et ml.,
1957

Bridle et •!.,
1979

"ickerlng »nd
Henderson, 1966


Brennisan et •!.,
1976









Devlin at el.,
(982
Fathead oinnow             25      SU
  (Pimephales proaetas)
Fathead Blnnov             25      SU
  (Plmephales proaelas)
16.3
(37.0
 to
59-1)
56.0
(11.7
 to
67.1)
16.3
( 37.0
 to
59.1)
                                                 la
                                                67.D
31.3
(3?.8
 to
15.9)
u?.3
(33.5
 to
5 J.',)
Tests were conducted
In soft water.
                                                      Teits were conducted
                                                      in hfird  wAler.
Pickering and
  Henderson, 1966

-------
TABLE 17-1 (cont.)
Species
Blueglll aunflsh
(Lepoaia nacrochirus)


Blueglll sunfiah
(Lepogia siacrochlrua)


Guppies
(Poecilia retlculata)


Zebrafiah
(Brachydanlo rerio)

>
)
i
Hedaka
(Oryziaa latlpea)

Heoaka
(Oryzlaa latlpes)


Coho salaion fry
(Cncorhynchus klsutch)
MA H INK
Coho sal non
(Onoorhynchus kiauteh)


Pink salmon fry
(Oncoriiynchug klsutch)


Tenp. Type 24 h
(«C) Test
25 SU 24.0
(18.9
to
30.5)
NR SU 16.6
(15.0
to
19.1)
25 SU 62.8
(55.0
to
73.7)
20+1 FU —




25+2 SU 80
(mean:
80)

25^2 SU 4-4


FH
FM

8 SU



12 SM 5.4
(4.4
to
6.5)
48 h 5072 h
24.0 —
(18.9
to
30.5)
13.3 12.7
(11.6 (11.5
to to
14.8) 14.5)
61.0
(52.8
to
71.9)
25 to
27



20 to 135
(oean=
63)

36


...
	 —

22.4 22.4



	 	



No Effect
96 I Concentration
24.0 	
(18.9
to
30.5)
12.7 10.0
(11.5
to
14.5)
59.3
(50.9
to
70.3)
	 —




23 to 110 O6
(Beans
54)

32


9.36
3.08

22.4 10







Reported
Concentration
Units
• mg/i



ppm



mg/l



tag/ 1




Bg/Jt



Eg/i


\ll/l
Hi/1

ppa



Pfxn



Conn en ta
Tests wcre-conduot-ed
in hard water.


Only these data
cited in U.S. EPA,
1980.

Tssts were conducted
in hard water.


Tests were conductsd
in closed aquaria
with dechlorlnated hard
tap water at a flow
rate of 6 i/h.
Range and oean of
LC values for dif-
ferent stage esbryoa

LC values for fry.
Th3Ul68 h. LC was
23og/l. 5°
Unparasitlzed
Parasitized

Tests were conducted
in artificial salt-
water (pH 8.1, 30°/oo
salinity).
Testa were conducted
according to methods
of Korn et al., 1979.

Reference
Pickering an
-------
                                                                     TABLE 17-1 (cont.)
Species Tenp. Typ« 2« h <18 h 5°72 h
( *C) Test
Pink saloon l| SM 	 	 	
(Oreorhynchus Iclsutch)


8 SM



12 SM



Striped bass 16 SM 7.3 	 —
(Morone saxattlls?


She«pshead Blr.,iow HR SU >277 >277 	
(Cyprinodon varlegatus) <«85 277
<«85
13
(5.0
to
35)


	






...


—

No Effect Reported
Concrntrallon .Concentration Cotraents Beferenne
Units
	 ul/l Tests were conducted Korn et al., 1979
with saloon fry
acclloated to 28°/oo
seawater at dlf-
	 ferent temperatures.



	



— ul/t Tests were conducted Benvllle «nd
In 25°/oo salinity Kom, 1977
seawater with Juvenll*
fish.
J77 PPB Data only cited In U.S. EP», T978
U.S. EPA, 1980.
	 mg/l Tests were conducted Bard et al. ,
In 15J sallnaty sea- 1981
water win Jutenlle
fish.


28 ng/l Test was conducted teBlanc, 1980
with reconstituted
wel! water (hardness
72+b ag/l as CaCO ,
pH 7.0+0.2) in 3
containers sealed with
plastic wrap.
	 mg/l Test was conducted Brlngsiann and Kuhn,
In natural water { pH 1959
7.5, hardness ?1ll Eg/I).
9. .95 ppm Test was conducted Berry and
with distil' d uramraer, T977
                                          to
                                        21.63)
Harln«

Brine shrimp nauplli
  (Srtemla sal Ina)
«f<:.5    SU   33
                                                                                                      water.
Test was conducted
with artificial sea-
water.
                                                                                                    Price et al., 1971

-------
TH8L8 17-1  (con'...)
Species
Bay shrimp
.(Crogo franclscorua)



Shrlap
(Eualua spp.)










Or»«» snrlrsp
(Peeaeaonetas pugic')













Crass shrltrcp
(Pacnemoneles puglo)




Teisp. Type
(*C) Test
16 SH




« SM



8 SH



12 SM



20 SM



20 SM



10 SM


10 SM



20 SH


20 SH


2» h
12
CO
to
13)

...



...







20.2
(16.3
to
22,5)
17.2
(16,9
to
19.1)
37.6
(35.0
to
50.3)
38.1
(36.1
to
39-6)
30.6
(21.3
to
6H.5)
25. «
(te.8
to
tc«;o
•8 h '72 h 96 h
	 «.3
(3.1
to
5.8)

	 21.*
(19.5
to
23.5)
20.2
(17.9
to
22.8)
	 11.7
(13-1
to
16.6)
... — - ...



	 __. .„_



... ...


___ 	



— — —


.__ ... ...


Ho Effect Reported
Concentration Concnntratlon
Units
lA/t




vtn



\ilfl



nt/i



•*/l



•g/l



VR/l


«R/t



Bg/l


Of/I


Coraewnts
Tests w«r« conduetad
with 25°/oo


salinity aeavater.












»dtilts st 15°/roo
salinity.


Mults at 25°/oo
salinity.


Adults at 15°/oo
salinity.

Kdults at 25°/oo
anllnlty.


Lsrrae at 15°/oo
salinity.

Lanrse at 25 /oo
salinity.

Reference
B*n»llle and
Kopn, 1977



(Corn et al..



(Corn et *!.,



Korn et at,,



Fotera, 1975



Potera, 1975



Potera, 1975


Potem, 1975



Poters, 1975


Potera, 1975








1979



1979



1979

























-------
                                                                    TABLE 17-1 (oont.)
Species
Cras.i shrimp
(Palacwistcs puglo)
Mysld shrlpip
(Hysldopala bahla)


Dungsnos? crab
(Cancr. ea^lster)
Co - 
-------
0
20
80
80
100
100
0
30
80
90
100
100
0
40
100
100
100
100
toluene concentrations.  Because the test containers were vigorously aerated, it
is probable that the actual toluene concentrations decreased rapidly during the
exposure period.    It  was  also observed  that  the  turbidity  of  the  toluene
solutions decreased from  150  to 100 ppra over  the  96-hour axposwre period.  At
concentrations of 560 ppm and  below,  all  fish appeared to  be  unaffected.  The
remainder of the test results are presented below:

          Concentration                   Percent Mortality
              (ppm)                        21 h     18_h
              < 560
              1,000
              1,600
              3,200
              5,600
             10,000
     Pickering and Henderson  (1966)  investigated the  acute toxicity of toluene
to   fathead  minnows   (Pimephales   promelas),   bluegill  sunfish   (Lepomis
tcacrochirus), goldfish  (Carassius  auratus),  and  guppies  (Lebistes reticulatus
- Poecilia reticulata).  The length and weight of the  fish  used  for testing were
3.8 to 6.U cm and 1 to 2 g for the first 3 species and 1.9 to 2.5 cm and 0.1 to
0.2 g for guppies.  Each  test utilized 10 fish per concentration or control in
either 10 S, (minnows,  sunfish, goldfish) or 2  I  (guppies) of soft water (pH 7.5,
alkalinity 18 mg/JL, EDTA hardness 20 mg/JZ,)  made by mixing 5 parts of hard natural
spring water with 95  parts  of  distilled  demineralised  water.   In  addition,
fathead minnows were tested (10 fish/concentration) in the  hard  spring water (pH
8.2, alkalinity 300 mg/£, EDTA  hardness 360 mg/£.)  to  investigate the effect of
these water  characteristics  on toluene toxicity.  All  tests  were conducted at
25°C.  The test solutions were not aerated, and dissolved oxygen concentrations
were measured but  not  reported.   The 24,  48,  and 96-hour LC    value? and their
95% confidence limits,  as calculated by the moving average-angle  method of Harris
(1959)   using   initial  nominal  toluene  concentrations,   are  presented  in
Table 17-1. The 96-hour LC,-n  values  increased  in the  order of  bluegill sunfish
(21.0 mg/£), fathead minnow (34.3 mg/£  in soft water, 12.3 mg/j(, in hard water).
goldfish (57.7 tng/H),  and  guppies  (59-3 mg/Jl).   The 96-hour LC5Q for fatt-ead
minnows in soft water was not significantly different from the  96-hour LC5Q for
the same species in hard water.  Comparison of the 95% confidence limits of the
96-hour  LC  n values in soft  water for the 4 species indicated that  the LC_0
values were  not  significantly different  between father! minnows  and  bluegill
                                      17-9

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aunfish or  between  goldfish and  guppies.   Both  fathead minnows  and bluegill
sunfish had 96-hour LC   values  significantly lower than goldfish and guppies*
The 96-hour LC^g was not significantly different from the 24-hour LC5Q  for any of
the species tested in soft water.
     Replicate  flow-through  acute  toxicity   tests  were  also conducted  with
fathead minnow embryos,  1-day-old  protolarvae,  and 30-day-old larvae by Devlin
et al. (1982).  The 96-hour LC    ranged between  18 and 31 mg/K, for  30-day-old
fish,, between  25  and 36 mg/S,  for protolarvae,and  between  55 and  72 mg/X, for
embryos.   Embryos were significantly more resistant that the other life stages.
     Static acute  LC  values for bluegill sunfish have also  been reported by the
U.S. EPA  (1980).  The 24, 48,  12, and 96-hour  LC5Q values were  16.6, 13-3,  12.7,
and  12.7 ppm,  respectively.   No  effects  were  observed at  or  below 10 ppm.
Additional information concerning  these tests was  not available.
     Berry (1980)  mentioned that the upper non-lethal toluene concentration for
bluegill  sunfish  (Lepomis macrochirus)  was  8.7 mg/!l.   The duration of exposure
and lowest lethal concentration.were not specified.
     Bridie  et al.  (1979)  and Brenniman  et  al.   (1976) also  investigated the
acute toxicity of toluene to  goldfish.   Bridie et al.  (1979)  used goldfish of
slightly greater  weight (mean 3.3 g,   range  2.3   to  '1.3 g)  than  Pickering and
Henderson (1966) to determine  the static 24-hour LC^.   In this test,  6 fish per
concentration  were  exposed without  aeration  to  a toluene  series in 25  I of
tapwater that had  a  pH of 7-8 and contained (in milligrams per liter):   Cl~ = 65;
N02"= 0;  N03" = 4; SC^2'  = 35;  P0^3~  = 0.15; HC03~ = 25;  Si02 = 25; NH^ = 0; Fe =
0.05; Mn  = r   ^a2* =  100;  Mg2"1" =  8;  and alkali  as Na4"  =  30.    The toluene
concentration  was measured  at the beginning  and  end  of the test.  The 2^-hour
LC,-,,, obtained by interpolation  from a  graph  of the logarithm of concentration
versus percent mortality, was..58 mg/Jt, which is the same as  the 24-hour LC   for
goldfish  reported by Pickering and Henderson  (1966).
     Much larger goldfish (length, 13 to 20 cm; weight,  20  to 60 g) were used by
Brenniman et al.  (1976)  to  determine the acute toxicity of toluene under flow-
through exposure conditions.  The LC_0 values  were determined by exposing 6 fish
per 38 i  aquarium to three  toluene concentrations  (and  a control) in  dechlorin-
ated soft tapwater  (methyl  orange  alkalinity  =  34  ppm as CaCO_; phenolphthaline
alkalinity  =  37 ppm as  CaCCK;  total  harness  =  80 ppm  as  CaCO^;   calcium -
21.6 ppm;  magnesium = 5.3 ppm;  SiO- = 8  ppm; chromium - <0.002 ppm; pH  7.0 + 0.3;
                                     17-10

-------
temperature 17  to 19°C) at a  flow rate  calibrateJ  to renew  the  test chamber
volumes every  1.5  hours.   This flow  rate was sufficient to maintain  dissolved
oxygen concentrations at _>7  ppm and to maintain constant toluene concentrations,
as measured by continuous monitoring at 210 ntn by spectrophotometer. The 24, 48,
72, and 96-hour  LC™ values,  calculated  by  probit analysis.,  were 41.6,  27.6,
25.3, and 22.8 ppra,  respectively-     'Ithough most  of the fi ;h died during the
first 24 hours, the 96-hour LCr^ wag significantly lower than  the  24-hour LC,-0«
These  LC,_0  values  are somewhat  lower  than those  reported  by  Pickering  and
Henderson (1966)  and Bridie et al.  (1979) for goldfish tested under  static condi-
tions.  In addition, the LC _ values  reported by Pickering  and Henderson (1966)
did  not  decrease significantly  from  24  to  96 hours.   These  differences  are
                                   j
probably due to a rapid decline in  the toluene concentration through evaporation
in the static tests in contrast to constant toluene concentrations in  the flow-
through  test.   Brenniman  et ai.  (1976)  continued  their  flow-through exposure
test  for  30 days,  at  which  time  the LC(-0 had decreased  to  14.6 ppm.   These
results emphasize the fact  that static acute  toxicity testa  may seriously under-
estimate the  acute  toxicity of toluene and  that  chronic effects  may  occur at
concentrations that are considerably  lower than those that  cause acute effects.
     Juhnke  and  Ludemann  (1978)  investigated  the  static  acute toxicity  of
toluene to the ide (Leuciscus ijdus  melanotus)  using comparable  procedures in two
different laboratories.   The  toxicity  tests were  conducted  according  to  the
methods of  Mann (1975, 1976),  i.e.  48  hours of  exposure  with  10 fish  (1.5 +
0.3 g, 5 to 7 cm) per concentration in tapwater (pH 7-8, hardness 268 + 54 mg/J!.)
at 20 +  1°C.   The 48 hour  LC  (Q% mortality). LC__, and LC.00 (100% mortality)
values determined at each laboratory were as follows:

                                   48 Hour Lethal Concentration Values  (mg/i.)
                                      T r-             ip             f ^
                                      "°0              50           ""100
          Laboratory  i                 72             70              88
          Laboratory 2                365            422             470
     Although it  was stated that  those  tests were  conducted  under comparable
conditions, the results were clearly different.  The concentration that caus«d no
deaths of  fish  in laboratory  2 (365 mg/2,) was  about  4 times higher  than the
concentration that killed all  fish in laboratory 1  (88 mg/i).   The authors did
not discuss the reasons for the difference in results.
                                     17-11

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     Sloof (1978,  1979) reported  that  the 48-hour LCcn of toluene  to  zebrafish
                                                     50
(Brachydariio rerio)  waa 25 to  27 mg/l.   This  test was  conducted  under  flow-
through  (6  &/hr) exposure conditions  using 10 fish  per concentration in  10 £
sealed aquaria and dechlorir.ated tapwater  (20 ^ 1°C; pH 8.0 +  0.2;  hardness  180 +
1.8 mg/K. as CaCO  ).
     The acute effects  of  toluene on parasitized and unparasitized coho salmon
(Oncorhynchus kisutch)  fry were studied by Moles  (1980).  The parasitized fry
were artificially infected before toluene exposure with glochidial larvae of the
freshwater mussel, Anodont? oregonensis.   Toluene exposure waa conducted  under
flow-through  conditions,  using  five measured  concentrations and  20  fisn per
concentration.   The  temperature and  characteristics of the water used were not
specified.     The  96 hour  LC     as  calculated  by  probit   analysis,  was
9.36 \iSL/i, (ppm)  for unparasitized fish and 3.08 \ii/i for fish parasitized with a
mean number of 69 glochidia  per fish.   The LC   values  were  significantly  dif-
ferent, indicating that parasitized  fish  were  less resistant to  the effects of
toluene.
     Stoss  and  Haines  (1978)  investigated the  effects of  static  exposure to
toluene on tht survival of fertilized eggs  and newly natched  fry  of the meclaka,
Cryzias la*ipes.  Groups of ten eggs or fry were exposed in loosely  capped  vials
'.sricaining  20 m£ of the exposure medium (synthetic  rearing medium:    pH   7.6;
akalinity 99 mg/£, as CaCO ) at 23  + 2°C.  Toluene concentrations were prepared by
                         j>
diluting a water-soluble extract  of  10 tat toluene/2, mec'ium.   In order  to deter-
mine the sensitivity of different stages of embryo development,  tests were  begun
with eggs of various ages  after fertilization.  Tests  with  fry were  all  begun
within  24 hours  after  hatching.,   Nominal  initial  toluene concentrations  were
used for calculation of LC5Q  valjes.  The  LC5Q values  for embryos  varied  with
length  of exposure and the age at time of  introduction.  The mean 24, 48, ar.d
96-hour LCc/- values for all ages of embryos  were 80, 63,  and  54 mg/£.  The  range
of LC    values  was 20  to  135  mg/fc at 48 hours and 23  to 110 mg/fc  at 96  hours
(Stoss, personal communication).   Karly  (£3.5 hours  old)  and late (^192  hours
old) embryos had significantly lower LC^n  values  at  each exposure period  than
embryos  of  Intermediate age  at  time  of  introduction.   The 24,  48,  96, and
168-hour LC.-,  values for fry were 44, 36, 32,  and  23 mg/Jl, respectively (Stoss,
           ylj
personal communication).   These  values  were  lower  than the maan  embryo  LC,-Q
values for the same  exposure period; however,  fry  LC    values were  greater  than
ths LCf-n values  for  the susceptible early and  late stage  embryos  and lower  than
                                     17-12

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most of the LC^Q values for intermediate stage embryos.  Stoas and Haines (1978)
also investigated  the sublethal effects of toluene on hatching time and induction
of  developmental  abnormalities.   These  sublethal  effects are discussed  in
Section  17.3-2.1.
     17.3.1.2.    MARINE FISH  —  Morrow et al.  (1975)  studied  the  effects  of
toluene  on young coho  salmon (Oncorhynchus kisutch) that had been acclimated to
artificial seawater (30 °/oo (parts per thousand) salinity; 8°C; pH 8.1) for up
to 2 weeks.  A  static  exposure technique was  used in which toluene  was  added
directly to exposure aquaria containing fish and  73 £, of seawater (
-------
conducted with 1C to  15  fry  per  concentration (<1 g fish/A water).   Fish were
added to the test containers after addition of an appropriate amount of toluene
in water stock solution.  The containers were not aerated until after the first
48 hours of exposure to minimize' evaporative loss.  Even so, analysis showed that
toluene decreased to nondetectable levels  by 72 hours at 12°C and by 96 hours at
8°c"and to 25% of the  initial concentration by 96 hours at 4°C. The 96-hour LC
values, estimated by probit analysis using initial measured concentrations  ex-
pressed as \ii/i  toluene  (= ppm),  were  6.4 at  4«C, 7.6 at 8°C, and 8.1 at 12°C.
The 95% confidence  intervals of  the  4°C and 12°C LC,-n  values  did  not overlap,
indicating  that  temperature affected  the toxicity  of  toluene.   There  was no
significant difference  between 24 and  96-hour  LCj-fs values because  almost  all
deaths occurred within the first  24 hours  of exposure.  The effect of temperature
may have been caused by greater sensitivity of the fish at the lower temperature
and/or by the longer persistence of toluene at the lower temperature.
     Thomas and  Rice (1979) used  the  previously described techniques  of Korn
et al.  (1979)  to determine  the  static 24~hour  LC-. of  toluene with somewhat
larger (1 to 2  g,  4.5 to 5.5 cm) pink  salmon fry at 12°C in  seawattr.  The 24-hour
LC    (and 95J  confidence interval) was  5.4  (4.4 to 6.5) ppm,  which is signifi-
cantly different  from the  96-hour LC^  value  of  8.1 ppm  (7.5  to 8.8) obtained
with younger fry at 12eC by Korn et al.  (1979).   The  reasons for this difference
cannot be determined from the information provided.
     A similar static exposure technique was used by  Benville -and Korn (1977) in
their study of the  acute toxicity of toluene to  juvenile striped  bass  (Morone
saxatilis)  in  aeawater  (25 °/oo  salinity,  16CC).   The test  was  initiated by
adding different amounts  of saturated toluene in water stock solution to the test
aquaria, each  containing 10  fish.  Toluene  concentrations were measured at the
beginning of the test and every 24 hours thereafter to the end of the test.  The
24 and  96-hour LC5Q values  were both 7-3 \ii/i  (ppm).   Almost all mortalities
occurred  within  6  hours.    The average  percent loss  of toluene  was 40$  by
24 hours, 53J by 48 hours, and  >99? by 72 hours.
     The only flow-through toxicity test with marine fish was conducted by Ward
et al.  (1981). The flow-through 96-hour LC^, based on measured concentrations,
was 13 rng/4  for Juvenile sheepshead minnows (Cyprinodon variegatus).  This value
was aauch lower than that obtained with static  tests.  The static 96-hour LC   for
similar fish was reported to be >277  <485  ppm in an unpublished U.S. EPA study
(1978, cited  in U.S.   EPA,  1980) and in  Ward et al. (1981).  Although toluene
                                     17-11

-------
concentrations were  not  measured in  the static  test,  the difference  in LC,-0
values is almost  certainly due  to  rapid loss of  toluene  from the static test
containers.
     17.3.1-3.  FRESHWATER  INVERTEBRATES — Berry and  Brammer (1977) investi-
gated the  acute static  toxicity of  toluene  to  fourth-instar  larvae  of the
mosquito,  Aedes  aegypti.   The  larvae  were  reared  fron  eggs  and  tested  in
distilled  water at  25 + 1°C.  For each of four replicate tests, duplicate groups
of 20 larvae each  were exposed  to 14 toluene concentrations. The mortality data
were pooled (160 larvae/concentration) to  calculate  the 24-hour  LCt-0 by probit
analysis.  Initial  exposure concentrations were determined  by gas-liquid chroma-
tography.  The 21-hour LC,-0 (± standard error) was 21.52  + 0.16  ppm.  The highest
concentration  (+  standard  error)  that  caused no mortality   over  the  24-hour
exposure period was 9-95 + 1.30 ppm.
     Berry (1980)  mentioned that the upper non-lethal toluene  concentration for
crayfish  (Orconetes  rusticus)  was  104.4 mg/d.   The  duration of  exposure and
lowest lethal concentration were not specified.
     The acute toxicity of toluene has also been  determined with the cladoceran,
Daphnia magna, by Bringmann and Kuhn (1959)  and by LeBlanc (1980).  Bringmann and
Kuhn (1959)  reported a 48-hour LCj-. of 60 mg/Jl.  This static test was conducted
with first instar  (<24 hours old) Daphnia magna in natural freshwater (pH 7.5;
hardness 214 mg/£.)  at 23°C.
     LeBlanc  (1980)  conducted static  tests with  first  instar (<24 hours old)
animals in deionized well water reconstituted to a total hardness of 72 j- 6 mg/R.
as CaCO, and a pH of 7.0  + 0.2 at 22 + 1°C.  Three groups of 5 daphnids each were
       j
exposed to each of  at least  five  toluene  concentrations and  uncontaminated water
in covered 250 mi  beakers containing 150  mS, of test solution.  The 24 and 48-hour
LCj... values (and 95$ confidence intervals),  based  on initial nominal concentra-
tions, were both 310  (240  to  420) mg/Jl.   The "no  discernible  effect concentra-
tion" was  28 mg/S,.   This  LC^ value  is  considerably higher  than that reported by
Bringmann  and Kuhn (1959).  The reasons for this difference cannot be determined
from the data provided.
     W.3.1.4.   MARINE  INVERTEBRATES --  Price  et al.  (1974)  determined the
static 24-hour LC,-,, of  toluene  to brine shrimp  nauplii   (Artemia  salina)  in
                  ;>U
artificial  seawater  (27.87 g/i,   Nad;  1.36 g/S,  CaSO^;   3.T g/£,  MgS04«7H20;
8.42 g/i MgCl2; 0.79 g/fc  KC1; 0.16 g/S, MgBr2«6H20) at 24.5'C. Groups of 30  to 50
newly hatched  brine  shrimp  were exposed  to  5 toluene concentrations in 100 mi
                                     17-15

-------
seawater-  The estimated 24-hour LC50, based on initial nominal concentrations,
was 33 mg/fc.
     Bay shrimp (Crago francisoorum) were shown by Benville and Korn (1977) to be
somewhat more  sensitive to toluene.   The  24-hour static LC,-ni  determined in
natural seawater (25 °/oo salinity) at 16°C, was 12 \i!L/l (ppm).  The 96-hour LC__
                                                                             50
for this species (4.3 uJl/Jl) was significa.itly lower than the 24-hour LC5Q (non-
overlapping T5/t confidence limits).   These values were calculated from initial
measured toluene concentrations.
     Korn et al.  (1979)  investigated  the  effects of  temperature  on the acute
toxicity of toluene  to another  genus of shrimp  (Eualus  spp.).   Shrimp (0.8 g;
6 cm long)  were acclimated  to  the test temoeratures  in  natural 26 to 28 °/oo
salinity seawater for 4 days and then exposed in  groups of  10 to  15 animals to a
series  of  toluene concentrations,  prepared  by  dilution of a  saturated water
solution.  The tissue loading in the  test containers was less than 1 g/i.  Mea-
surement by UV spectrophotometry showed that toluene concentrations decreased to
nondetectable levels by 72 hours at  12°C and by  96 hours  at 8°C, and to 25$ of
the initial concentration by 96 hours at 4°C.  The 96-hour LC,-n values, calcu-
lated  from  initial  measured toluene  concentrations,  were  21.4 \ii/i  at  4°C,
20.2 \ii/SL at 8°C, and 14.7 jii/fc at 12°C.  The 96-hour LC  Q values at  4°C and 8°C
were not significantly different  (overlapping  95$ fiducial limits)  from each
other,   but both were significantly higher than the 96-hour LC   at  12°C.  This
trend of greater toxicity at  higher temperatures was opposite to the relationship
found  by these authors for  pink salmon fry (Section  17.3.1.2.) and by Potera
(1975)  for grass shrimp (see below).  The reasons  for  this difference could not
be established but may have  been due  to some combination of effects of tempera-
ture on persistence  of  toluene  in water, altered  toluene  uptake and metabolic
rates,  and possible interaction of toluene toxicity and temperature stress.  The
authors concluded  that  temperature  affected the toxicity of toluene  to these
species of  shrimp and salmon  but that  it  would be impossible  to  predict the
effects of temperature change on the  toxicity of toluene to other species.
     Potera  (1975)  investigated  the  effects  of  temperature   (10  and  20°C),
salinity (15 and 25 °/oo), and life stage (larvae  and  adults) on the static 24-
hour LCj.  of toluene to the grass shrimp, Palaemonetca pugio.  The 24-hour LCj-g
values, based on measured  initial concentrations, ranged from 17.2 to 38.1 ing/S,.
                                     17-16

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     As shown by overlapping 95?  confidence intervals  (Table  12-1), there was no
significant difference  in  LC(-n values  between  adults and  larvae at  the  same
salinity and temperature, or between adults tested at the same temperature but at
different salinities.  The  LC5Q was significantly lower at 20°C, however,   than
at ip"DC  for  adults  tested  at either  15 °/oo or  25  °/oo  salinity.   The time to
produce narcosis in at least 50? of adult shrimp at 20°C was less than 30 minutes
at initial exposure  concentrations of 19.8 mg/JZ,  and greater.  Recovery of more
than 90$ of exposed  shrimp  could  occur if shrimp were transferred to clean water
after exposure to up to 30 mg/2, for 30 minutes.
     Potera  (1975)  also determined the 24-hour  LC™ for the copepod, Nitocra
spinipes,  at a  temperature of  20°C  and at  salinities  of   either  15 °/oo  or
25 °/oo.   The 24-hour LCc0  values  from replicate  tests were  24.4  at 15 °/oo
salinity and  74.2 mg/Jl  at  25 °/oo salinity.   These  values  were significantly
different  (non-overlapping 95? confidence intervals).   Potera (1975) suggested
that the lower salinity  may have  stressed the copepods, resulting in a lower LC
value.
     Neff  et  al.  (1976)  also determined the static  96-hour  LC,Q of toluene to
grass shrimp, Palaemonetes pugio.  This value, based on  initial nominal concen-
trations, was 9-5 mg/£,  which is lower than the 24-hour  LC    values reported by
Potera (1975).
     Caldwell et  al.  (1976) determined the  48  and 96-hour  LC   of toluene to
larvaL stages of  the dungeness  crab  (Cancer magister) under  flow-through expo-
sure conditions.  The 48 and 96-hour LC^ values were 170 and 28 mg/Jl, respec-
tively.
     Static  acute LCcn values  for mysid shrimp (Hysidopsis  bahia)  have  been
                     5U                            —     	 	
reported by the  U.S.  EPA,  (1980).  The 24 and 48 to 96-hour LC50 values were 64.8
and  56.3 ppm,  respectively.    The   "no  effect"  concentration  was  27.7 ppra.
Additional information concerning this test was not available.
     The  48-hour  static  LC    of  toluene  to  larvae   of  the  Pacific oyster
(Crassostrea  gigas) was reported to be  1050 mg/Jl (LeGore, 1974).  This  test was
conducted  with  filtered seawater (25.3 to  30.8  °/oo  salinity) at 20 to 21.5°C
using 30,000  larvae per exposure concentration.
                                      17-17

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17.3.2.   Sublethal Effects.
     17.3.2.1.   FISH  —  Very little  information  is available concerning the E jb-
lethal effects  of toluene  exposure  on  fish.   Morrow et al.  (1975)  studied  the
effects  of several aromatic hydrocarbons, including toluene, on the  'evela of Na+
and K* in  the  blood  of young coho  salmon (Oncorhynchus kisutch)  in seawater.
Static exposure to 30 ppm  toluene caused a small increase in these blood cations,
reaching a maximum at about 2 hours after beginning  exposure.   The Na+ concen-
tration returned  to  the control level  by 3  hours.   Blood K+  decreased  after
2 hours  but was  still elevated at ^ hours,  the  last sampling period.  The toluene
exposure concentration  of  30 ppm was  sufficient to  cause  some  mortalities  and
behavioral effects.   The  authors   suggested  that  toluene increased  membrane
permeability, particularly in the gills.  In the hypertonic  seawater medium, this
change would  result in ion  influx and water loss in the fish,  perhaps accounting
for the initial rise  in blood ion concentration.
     Brenniman et al. (1979) conducted a series of experiments to determine  the
effects of toluene exposure on blood gas physiology, hippuric acid content,  and
histopathology of goldfish  (Carassius auratus).  The fish used in these experi-
ments were exposed  to  two or  more toluene  concentrations  under  flow-through
conditions using dechlorinated tapwater.
     For the pathology study, groups of six fish were exposed for up to 30 days
to 0, 5,  10, and  21  ppm toluene (Brennim^n  et al.,   1979).  No  gross or micro-
scopic lesions  were  observed  in fish  during the first week of exposure.  After
the first week,  ascites  developed in 3 fish at  21 ppm and in 2 fish at  10 ppm.   In
exposed fish that survived 15 to 30  days, about 50J had a white epidermal exudate
of unknown origin, and some fish at  all toluene concentrations had gross lesions
in gill, liver, or gall  bladder.  Excessive mucus production in gills  occurred in
all fish at 21 and 10 ppm  and in 50? of the fish at 5 ppm.  Microscopic lesions
were found in gills  (fusion),  liver (decreased cytoplasmic nuclear ratio),  and
kidney  (tubular vacuolization)  of  many exposed fish buc   not in  control  fish.
Exposed fish did  not eat  food  and  had livers that  were paler  and smaller than
control  fish.
     For the blood gas study, groups of 3 or  ^4  fish were exposed for  H  hours to 0,
60, or 80 ppm toluene (Brenniman et  al.,  1979).  The  blood samples were analyzed
for pH,  percent oxygen saturation, partial pressures of carbon dioxide (pCQ ) and
oxygen (p  ), and bicarbonate.  The results are presented below:
         °2
                                     17-18

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Mean Values
Toluene Cone.
(ppm)
0
60
80
"°2
42.33
I6.25a
15.633
Rco2
11.50
23.25a
19.27
PH
7.56
6.90a
6.96a
0 -Saturation
2 (?)
'18.67
27.00a
20.33a
Bicarbonate
9-83
5.10
*4.17a
  p <  0.05 when compared to control.

     Toluene exposure  caused  significant changes  in  all  parameters (Brenniman
efc al., 1979).  The authors suggested that the decreased pn  , increased p,  ,  and
                                                        U f-\             \j U fj
resultant acid-base imbalance  may have been  due to lowered u  and CO- exchange at
the gills.   Two  proposed  mechanisms  for impaired gas  exchange vere  lowered
respiratory rate and gill  damage.   The former  mechanism is less likely because
sublethal toluene exposure has  been shown to increase the  respiratory  rate in
fish  (Sloof,  197C,  1979;  Thomas  and Rice, 1979).   The  latter mechanism is
supported by the authors'  observation that  toluene caused excess mucus produc-
tion and fusion of gill lamellae in gills.
     The whole-fish  content  of hippuric  acid  was measured  in  fish  exposed in
groups of 6 fish to 0,  5, 10,  or 21 ppm toluene for 96 hours (Brenniman et al.,
1979).  This experiment was conducted to determine whether  the fish were able to
metabolize  toluene  ultimately  to  hippuric   acid,   as   occurs   in   mammals
(Chapter 12.).  The  results,  presented below,  indicated  that hippuric  acid  was
elevated at all  the toluene concentrations tested and that this metabolic pathway
occurs in goldfish.
Toluene Concentration
(ppm)
0
5
10
21
Mean Hippuric Acid Concentration
(ppm)
1539-50
3608.67
3536. 67a
2829- 17a
a
 P < 0.05 when compared to control.

The pattern  of decreasing hippuric acid  concentration  with increasing toluene
concentration was attributed to increasing stress  and lower metabolic efficiency
                                     17-19

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as toluene concentration increased.  Hippuric acid was elevated above the control
levels, however,  even at the highest toluene concentration.
     The only other information available relevant to toluene  metabolism in fish
is provided by Ohmcri et al. (1975),  who investigated the comparative in vitro
metabolism of a toluene analog,  £-nitrotoluene,  by liver  homogenates of rats and
eels.  The species of eel was not specified.  Both species were able to metabo-
lize £-nitrotoluene (PNT) to £-nitrobenzoic acid  (PNB acid),  via oxygenation of
PNT  to  jg-nitrobenzyl  alcohcl  (PNB  alcohol),  to  £-nitrobenzaldehyde  (PNB
aldehyde), and finally to PNB acid.  The rate of the overall reaction  (PNT to PNB
acid) in eel liver,  however;   was only 3^$ (at 25°C) to 1)6$ (at 37°C)  of the rate
in rat liver.  The rate of formation of PNB alcohol from PNT in eel  liver was 29%
(at  25°C)  to  16$ (at 37°C) of  the rate  in  rat  liver.   This  step was the rate-
limiting step for the  overall reaction because the formation of PNB  acid from PNB
alcohol was faster in eels than  in rats.
     Thomas and  Rice  (1979)  measured the effects of flow-thrcugh toluene expo-
sure on the respiratory rate and oxygen consumption of pink salmon  (Oncorhynchus
gorbuscha) fry at two temperatures (4°C,  12°C) in  seawater. The fish were placed
in sealed  chambers  fitted  with a water  inlet and outlet, mesh electrodes (for
measuring opercular breathing rate),  and oxygen electrodes (for measuring oxygen
concentration of inflowing and outflowing water).   After  determining the S^-hour
LCj-,.  (5.38 ppm), the  authors  exposed  fry  to  several  toluene concentrations,
expressed  as  percentages  of  the  LC,-,..    Significant  increases  in  opercular
breathing rate at 12°C occurred  at exposure concentrations of  91) and 69? of the
LC50,  but  not at ^45 or 30$ of the LC5p.  The  breathing  rate remained elevated
throughout the 15-hour exposure  period only at 9^$ of the LC,_0, at which concen-
tration 6 of 23  fish  died.  The  breathing rate at a toluene exposure concentra-
tion  of  69$ of  the LC_n  reached a maximum at  3  hours  and  returned  to control
level  by  15 hours.  Additional  experiments  showed that exposures to  71$ of the
LCc-Q increased oxygen consumption.  The percent increase  in both oxygen consump-
tion and breathing rate was greater at ^°C  than at 12°C.  The  authors suggested
that these effects were due to the energy requirements for metabolism of toluene
and  that this requirement was  greater at the lower temperature.  The threshold
for an effect on breathing  rate  at 12°C was estimated  to be about 16$ of the LC5Q,
or about 2.5 ppm.
     Sloof  (1978,  1979) conducted similar  experiments  to  determine  the sensi-
tivity of a biological  monitoring  system  using  fish respiratory rates  as  an
                                      17-20

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indicator of water pollution by toluene and other chemicals.  Adult rainbow trout
(mean weight  56 g) were acclimated  to dechlorinated tapwater at 20  + 1°C and
tested individually in  sealed  flow-through  chambers equipped  with  stainless
steel mesh electrodes for measuring breathing rate.  After the normal breathing
rate for a fish  over  a 3 day period had  been  determined,  toluene contaminated
water was added continuously  and  the breathing rates were monitored over a period
of 1*8 hours.  Measurements were  taken  at the same time  of  day  during the pre-
exposure and exposure periods,  A toxic effect was considered to have occurred if
the respiration frequency of  at  lea.it  75% of the test fish exceeded the prede-
termined individual normal frequencies  measured  at the same hourly interval.  The
lowest toluene  concentration that caused an increase in  respiratory rate was
2.5 rae/JU  This concentration is  identical to the estimated threshold concentra-
tion for an effect on breathing rate in pink salmon  (Thomas ana Rice,  1979).
     Leung and Bulkley (1979)  investigated the effects of 100 \ii/i toluene on the
rate of opercular movement by 8-day-old embryos  of the Japanese medaka, Oryzias
medaka.  The basal (unexposed) rate was determined for each of three embryos and
then toluene was added  to the culture medium to obtain a nominal concentration of
100 112,/i,.   The  rate  was  then   determined  for  each  embryo at  about 5 minute
intervals  for   40  minutes.    The  average  rate  before  exposure  w&s  zero
movements/minute.  The average of  8 counts  (each 1 minute long) over 40 minutes
after beginning  exposure was 2.28  movements/minute.  The standard deviation was
so great, however,  that this increase  was  not  statistically significant.
     The sublethal effects of toluene  on  medaka  were also investigated by Stoss
and Haines (1979).  The exposure techniques and lethal effects reported by these
authors  have  been discussed  in Section 17*3-1-1 •   Static exposure of eggs to
initial nominal concentrations of  11 and  82 mg  toluene/2, resulted in a signifi-
cant delay in time to hatching and a decrease in the proportion of embryos that
hatched successfully.  Exposure  to 41 ng/JZ.  and  greater caused numerous develop-
mental abnormalities,  including  disruption of  cell  cleavage  patterns,  defor-
mation of eyes, appearance of isolated  tlood islands  in  the circulatory system,
and abnormal  heart structure, t?il flexures,  and  visceral organ formation and
placement.  No abnormalities were observed in embryos exposed to 16 mg toluene/S,.
     Subchronic embryolarval toxicity  tests have been conducted with  freshwater
fathead minnows  (Devlin  et al.,  1981)  and  saltwater sheephead minnows (Ward et
al., 1981).  Devlin et al. (1982)  exposed embryos under  flow-through  conditions
to  H  to  15 mg/ii,  toluene for  36 days.   Larval growth  was  the  most  sensitive
                                      17-21

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indicator of toxicity, with significant inhibition of larval growth occurring at
toluene concentrations  as  low aa 6 mg/£..   No  effects were observed at 4 mg/JL.
The maximum acceptable  toxicant  concentration  (MATC) was,  therefore, between 4
and 6 mg/J, for this species.
     Ward et  al.  (1981) conducted flow-through  toxicity tests  with embryos of
sheepshead minnows exposed to  1  to  19  mg/J,  toluene for 28  days after hatching.
Exposure  to 7.7 mg/fc  and  greater  caused a  significant  decrease  in hatching
success and survival of Juveniles.  There were  no effects on growth of surviving
fish.   As a  result,  the MATC was  >3.2 <7-7 mg/Z..   The 96-hour  LC   for this
species was between 277  and 485 ppm (Section 17.3.1.2.). The ratio between acute
and sub-chronic toxicity was between 36 and  152,  indicating that chronic effects
occur at concentrations much lower than  acute  effects.
     In  summary,   the lowest  toluene   concentration shown  to  cause sublethal
effects in fish was 2.5  ppm, the concentration that caused an increased breathing
rate in trout (Sloof, 1978,  1979) and salmon (Thomas  and Rice,  1979).  This va]ue
is .somewhat  below the  lowest  acute LCj-r, value  reported  for any  fish  species
(3.08 ppm for coho salmon, see Table 17-1).  An  embryo-larval test with sheeps-
head minnows  (U.S.. EPA,  1980)  showed  that subchronic toxic  effects occurred at
7.7 ppm but not at 3-2 ppm and that  the ratio of  the  acute LC    to the subchronic
MATC for this species was between 1.7  and 4.0.   Another embryo-larval test with
fathead minnows (Devlin et al., 1982)  showed that subchronic effects occurred at
6 mg/£, but not at  4 mg/H, and  that the  ratio of the acute LC   to the subchronic
                                                           -'U
MATC was between 3 and  18.  Although acute-chronic ratios may vary greatly among
species, this information suggests that  chronic  toxic effects may occur in coho
salmon and other sensitive species at  concentrations well below 3 ppa.
     17.3.2.2.   INVERTEBRATES — Berry et al.  (1978)  conducted  a series  of
experiments to determine the effects of 24 hours  of exposure to sublethal concen-
trations  of  water-soluble  fractions (WSFs) of gasoline, benzene,  xylenes,  and
toluene on  oxygen  consumption by fed  and unfed  larval stages  of the mosquito,
Aedes aegypti.  Control experiments  with untreated animals showed that there was
no significant difference in 0  consumption  between fed and unfed larvae.  Treat-
ment with the WSF of  1 mi/i gasoline, however  caused  an  increased 0^ consumption
in fed, but not unfed,  larvae  relative  to untreated  controls.  Treatment of fed
larvae with individual  WSFs of benzene  (1 mi,/£),  xylenes (0.3 m£/£,), or toluene
(0,1 to 0.5 mA/H)  had no effect on  0_ consumption relative to fed controls.  A
WSF mixture of benzene,  xylenes, and toluene and a mixture of benzene and toluene
                                      17-22

-------
(0.2 mil/jt, for  each compound) caused  significant  increases in  0- consumption.
Exposure to a WSF mixture of benzene and xylenes or toluene and xylenes (0.2 mJl/£
for each compound) had no effect.  The authors also conducted experiments on the
uptake of %-labeled toluene in fed and unfed animals, as well as uptake of ^H-
toluene by  fed larvae in the presence  or absence of  benzene  (Section  15.3.).
Maximum -^H-toluene counts were  equal  in fed and unfed larvae,  but were reached
more quickly (1 hour versus ^ hours) by the fed animals.  The ^H-toluene counts
in larvae, expressed as the  percentage of the initial water counts, were greater
in the benzene and toluene mixture than in the solution  containing  toluene alone.
The authors concluded that the effects of gasoline on 0  consumption were due to
the enhanced uptake and  synergistic effects of  toluene and benzene,  two of the
major aromatic components of gasoline.  They also  suggested that the presence of
food accelerated  the uptake of  toluene  through  absorption of toluene  to the
consumed food particles.
     Blunuo (1978) investigated the effects of toluene on the swimming activity
and survival  of  barnacle  (Balanus  eburneus) larvae.    Groups  of larvae  were
exposed for 1 hour in specially constructed tubes to 10,  20, 30, 40, 50, 60, 70,
80, and 90? of th-3 water soluble fraction  (WSF) made by saturating seawater wjth
toluene.  The tubes were designed  so that  actively swimming  photopositive larvae
would be attracted to light at the top  of the tube.  After 1  hour of exposure, the
inactive larvae  were  collected  from the bottom  of the tubes and  stained with a
vital dye (neutral red)  to determine percent mortality.  The remaining portion,
containing  the active larvae, was then collected  and counted.  The interpolated
concentration  that immobilized 50$ of  the  larvae was  12.5?  of  the WSF.   All
larvae were immobilized  at 30? WSF and higher.   About 33-1/3$ of the larvae were
immobilized at  10? WSF,  the lowest concentration  tested.  The percent mortality
of the immobilized larvae ranged from about 3? at 10? WSF to a maximum of 12$ at
90? WSF.   The author also measured the  effects of WSFs  that had been  aged in
covered  containers for  1 day  in a refrigerator  or  exposed  to  air for  up to
3 days.  The percent WSF that immobilized 33-1/3? of the larvae was  10? in the
fresh solution,  37.5? in the refrigerated  solution,  and 90?  in  the evaporated
solution.   Additional  experiments  showed that aeration  of  the  WSF for 6 hours
lowered the toxicity to  the same extent as 3 days  of exposure to  air.
     Bakke  and Skjoldal  (1979) investigated the effects  of  toluene on activity,
survival, and physiology of the isopod,  Cirolana boreaJis.   For determination of
median effective .times   (ET,-ni  partial or complete narcotization  as  endpoint),
                                      17-23

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groups of 15 isopods were exposed in  duplicate to nominal initial concentrations
of 0, 0.0125, 1.25, 5.7, 12.5, 25, and 125 ppm  toluene for ^  days.  The exposure
medium  (33-5  to 3H.5 °/oo salinity  seawater  at 8  to 10°C) was  changed  every
2 days.  The interpolated or extrapolated ET50 values were as follows:
                  Toluene
               Concentration                   /r1 50  .
                  ippmj	                   (hours)
                  0
                  0.0125
                  1.25
                  5.7                            MOO
                 12.5                             69
                 25                               26
                125                                3

     No effects on activity were observed  in animals exposed to 1.25 ppm or less
(Bakke  and  Skjoldal,  1979).    The authors also  investigated the  recovery  of
isopods after exposure for varying periods to 12.5 or  125 ppm toluene.  Exposure
to  125  ppm  for  1 hour  caused  complete inactivity,  but all  animals recovered
within  12 hours after transfer  to clean water.  Exposure for 2 or more hours to
125 ppm caused  partial  or  complete mortality.   All  isopods could recover after
exposure to 12.5 ppm for 30 hour-s but not  longer.  Additional experiments showed
that  there  was no  significant  effect of  M days  of exposure to  up  to 5.7 ppm
toluene on oxygen consumption, ATP concentration,  or energy charge.  Exposure to
12.5 ppm resulted in a progressive decrease in ATP level and energy charge over
8 days  of  exposure, at  which time  all  organisms had  died.   Exposure to  the
rapidly lethal concentration of 125 ppm toluene showed no effect on ATP level or
energy charge *  These results  Hith 12.5 and 125 ppm were essentially the same as
those reported  by  the authors  in  a  previous  paper  (Skjoldal and Bakke,  1978).
Bakke and Skjoldal  (1979) concluded  that  the effect of toluene  on activity was
much  more  sensitive  as an indicator of  sublethal  toluene toxicity  than  its
effects on respiration, ATP level, and energy charge.
     In  sucmary,  the  lowest  toluene concentration  shown  to cause sublethal
effects in invertebrates was  5.7 ppm,  the concentration that caused narcotiza-
tion  of isopods (Bakke  and Skjoldal,  1979).   This  concentration  is somewhat
higher than the 96-hour LCc0 of 1.3 ppm for bay shrimp  (see Table  17-1) reported
by Benville and Korn (1977).  The latter concentration is the lowest reported to
have toxic effects on freshwater- or marine invertebrates.  Although the chronic
toxicity of toluene to aquatic invertebrates has not been studied,  it  is probable
                                     17-2'!

-------
that chronic effects could occur in sensitive invertebrate species at concentra-
tion  below  Jf.3 ppm.   This conclusion  is supported  by  the fact  that  chronic
effects in fish occurred at concentrations well below the acutely toxic concen-
trations (Section 17.3-2.2.).

17.1.  REFERENCES

BAKKE, T.  and SKJOLDAL,  H.R.    (1979).   Effects  of toluene on  the  survival,
respiration,  and  adenylate system  of  a  marine  isopcd.    Mar.  Pollut.  Bull.
10 CO; 111-115.

BERRY, W.O.  and  BRA>*ffiR,  J.D.   (19<'7^.   Toxicity  of  water-soluble  gasoline
fractions to  fourth-instar larvae of the mosquito Aedes aegypti L.   Environ.
Pollut.  13(3): 229-234.

BERRY, W.O.,  J.D.  ERAFtffiR,  and  D.E.  BEE.    P978).   Uptake of  water-soluble
gasoline fractions and their effect on  oxygen  consumption  in aquatic  stages of
the mosquito  (Aed'es aegypti L.)   Environ. Follut.  15: 1-22.

BLLFNDO, R.  (1978).  The toxic  effects  of the  water  soluble fraction  of No. 2
fuel  oil  ;«id of  three aromatic  hydrocarbons  on the behavior and  survival  of
barnacle larvae.  Contrib. Mar. Sci.  2]_: 25-37.

BENVILLE, P.E., JR. and KORN, S.  (1977).  The acute  toxicity of six monocyclic
aromatic crude oil components to  striped bass (Morone saxatilis) and bay shrimp
(Crago franciscoruen).  Calif. Fish Case.  63(^0; 20*4-209.
BERRY, W.O.  (1S80).  A comparative study of the uptake of toluene by bluegill,
sunfish Lepomis marcochirug and crayfish Orconectes rusticus.  Environ. Pollut.
80: 109-119.

BRENNIHAN,  G,,  HARTUNG, R. and  WEEER,  W.J., JR.   (1976).  A  continuous  flow
bioassay methoo to evaluate the effects of outboau^cS motor exhausts and selected
aromatic toxicants on fish.  Water Fes.  10(2): l65-'i69.
                                     17-25

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BRENNIMAN, G.R., ANVER, M.R., HAflTUNG, R. and ROSENBERG, S.H.  (1979).  Effects
of outboard motor exhaust emissions on goldfish (Carassius auratus).  £. Environ.
Pathol.  T6xicol.  2(6): 1267-1281.

BRIDIE, A.L., et al.   (1979).   BOD and COD of some petrochemicals.  Water Res.
_13: 627-630.

BRINGMANN, G. and KUHN, R.  (1959).  The toxic effects  of waste water on acjuatic
bacteria, algae, and small crustaceans.   Gesundheis-Ingerieur.  60: 115.  (Cited
in McKee and Wolf,  1963).

CALDWELL, R.S., CALDARONE, E.M. and tiALLON, M.H.  (1976).  Effects of a Seawater-
Soluble Fraction of Cook  Inlet Crude Oil and Its Major Aromatic Components on
Larval Stages of the Dungeness  Crab,  Cancer magister dana.  In:  Fate and Effects
cif Petroleum Hydrocarbon jln Marine Organisms and Ecosystems.  Pergamon Press, NY.
pp. 210-220.

DECANI,  J.G.    (19^3).   Studies of the  toxicity of ammunition  plant  wastes to
fishes.  Am. Fish Soc. Trans.   73; 45-51.

DEVLIN, E.W., 3RAMKSR, J.D. and PUYEAR, R.L.   (1982).   Acute toxicity of toluene
to three age groups of fathead  minnows,   bull. Environ. Contain. Tox.  29:  12-17-

FUNASAKA,  R.,  OSE,  Y. and SATO, T.   (1975).   Offensive  odor of fish  from the
Niagara-  River.    III. Aromatic  hydrocarbons  as  one  of  the  offensive-odor
substances.  Eisei  Kagaku  21(2): 93-100.  Take from:   Chem. Abst.  £3:  173356n,
1975.

HARRIS, E.K.   (1959).  Confidence limits for the LC,_0  using the moving average-
angle  method.   Biometrics.  15:  t»2H-'432.   (Cited in  Pickering  and  Henderson,
1966).

JUHNKE,  I. and LUDEMANN,  D.    (1978).   Results of research with 200  chemical
compounds  on acute  fish  toxicity with the golden orfe  test.  £. F_.  Wasser-und
Abwasser-Forschung.   '< 1 (5);  161-16^.
                                      17-26

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KORN, S., MOLES,  D.A.  and RICE, S.D.   (1979).   Effects of temperature on  the
median tolerance limit of  pink salmon and shrimp exposed to toluene, naphthalene,
and Cook Inlet crude oil.   Bull. Environ. Contam. Toxicol.   21(4-5):  521-525.

LeBLANC, G.A.    (1980).   Acute toxicity  of  priority pollutants to water  flea
(Daphnia magna).  Bull. Environ. Contam. Toxicol.  2_U:  684-691.

LeGORE, R.S.  (1974).  The Effect of Alaskan Crude Oil  and  Selected Hydrocarbon
Compounds on  Embryonic Development  of  the  Pacific  Oyster Craasostrea gigos.
Ph.D. Dissert. Univ.  Wash.  190 pp.  (Cited in  U.S.  EPA,  1980).

LEUNG, T.S. and BULKLEY,  R.V.   (1979).   Effects of petroleum  hydrocarbons on
length of incubation and hatching success  in the  Japanese Medaku.  Bull. Environ.
Contam.  Toxicol.  23: 236-213.

McAULIFFE, G.D.   (1976).  Dispersal and Alteration of Oil Discharged  on a Water
Surface.  In:  Fate  and Effects of Petroleum Hydrocarbons  in  Marine  Ecosystems
and Organisms.  D.A. Wolfe, Ed.  London:  Pergamon Press,   pp. 363-372.

MACKAY, D.  and  WOLKOFF, A.Q.   (1973).   Rate of evaporation  of low-aoiubUity
contaminants from water bodies  to  atmosphere.  Environ. Sc 1.  Technol.   T_:  611.
(Cited in Syracuse Research Corporation,  1980).

MACKAY, D.  and  LEINONEN,  P.J.   (1975).   Rate of evaporation  of low-solubility
contaminants  from   vater  bodies   to   atmosphere.    Environ.   Sci.   Technol.
J9: 1178-1180.

MANN, H.  (1975).  The golden orfe test:  German  proposal for testing  the action
of chemical compounds on fish.  Vom Wasser.  44:  1-13.

MANN, H.   (1976).  Comparative acute toxicity  testing  of  water pollutants  and
wastswater with the golden orfe fish test:  Experimental results from  three  ring
te.-sts.  Z. £.  Wasser-und Abwasser-Forsehung. _9_: 105-109.

MOLES, A.   (1980).   Sensitivity of  parasitized  Coho salmon  fry to  crude  oil,
toluene and naphthalene.  Am. Fish Soc., Trans.   109(3): 293.
                                     17-27

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MORROW, J.E., GRITZ,  R.L. and KIRTON, M.P. (1975).   Effects of some components of
crude oil on young Coho salmon.  Copeia.  2i 326-331.

NEFF,  H.J.,  ANDERSON,  J.W.,   COX,   B.A.,  LAUGHLIN,  R.B.,  ROSSI,  S.S.  and
TATEM, H.E.  (1976).  Effects of petroleum on survival, respiration, and growth
of, marine animals.  Proc. of the Sywp. Amer. Univ., Washington, D.C.
   \

OGATA,  M.  and M1YAKE, Y.   (1973).   Identification  of substances  in petroleum
causing objectionable odor in fish.  Watjer Res.  T_:  1^93-1 SOU.

OHHOSI,  S.,  et  al.    (1975).    The metabolism and  accumulation  of petroleum
components in fish, the side chain oxidation of p-nitrotoluene and p-nitrobenzyl
alcohol  in liver  hesaogenates  of  the rat  and  eel.   Physiol.  Chen.  Physics.
7: .177.

PICKERING,  Q.H.  and HENDERSON,  C.   (1966).  Acute  toxlcity  of acme important
petrochemicals to fish.  .J. Water Pollut. Contr. Fed.  58(9): TJ19-1^29.

POTERA, G.T.   (1975).  The effects of benzene, toluene, and  ethyl  benzene on
several  important  members  of  the  estuarine  ecosystem.    Piss.  Abstr.  *3-
36(5): 2010.

PRICE,  K.S.,  WAGGY,  G.T.  and CONWAY, R.A.   (WO.   Brine  Shrimp  bioassay and
seawater BOD of petrochemicals.  .J. Water Pollut. Cont. Fed.  ^6(1): 63-77.

SHELFORD, V.E.  (1917).  An experimental study of the  effects of gas waste upon
fishes with special  reference  to stream  pollution.   Bull. 111.  State Lab. Nat.
Hist.   11:   .

SKJOLDAL, H.R. and BAKKE,  T.  (1978).  Relationship between ATP and energy charge
during lethal metabolic stress  of the  marine isopocl Cirolana borealis.  J_. Biol.
Chem.  253(10); 3355-3356.
                                     17-28

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SLOOF,  W.    (1978).    Biological  Monitoring  Based  on Pish  Respiration for
Continuous Water Quality Control.   In:   Aquatic Pollutants.   Transformation and
Biological Effects.   0.  Huzinger and S. Safe,  Eds.,  Pergamon Press.  Vol.  1,
pp. 501-506.

SLOOF, W.  (197S).  Detection limits of a biological monitoring system  based  on
fish respiration.  Bull. Environ. Contain. Toxicol.   23(^-5):  517-523.

STOSS, F.W. and HAIHES, T.A.  (19?9).  The effects of  toluene on embryos and fry
of the Japanese medaVca Oryziaa latipes with a proposal for rapid determination  of
maximum acceptable toxicant concentration.  Environ. Pollut.   20(2):  139-1*18.

THOMAS, R.E.  and RICE,  S.D.  (1979).   The effect  of exposure temperatures  on
oxygen  consumption and opercular breathing rates of pink salmon  fry  exposed  to
toluene,  naphthalene,  and  water-soluble fractions  of Cook Inlet crude oil and
No. 2 fuel oil.  Mar.  Pollut.  T9_: 39-52.

U.S. EPA  (U.S. ENVIRONMENTAL  PROTECTION AGENCY)  (1980).  Ambient Water Quality
Criteria  for  Toluene.   Publication  No. EPA  '410^5-80-075.    U.S. Environmental
Protection Agency. Washington, DC.

WALLEN, I.E., GREEN, W .C. andLASATER,  R.  (1957).   Toxicity to Gambusia affinis
of   certain   pure  cneoiicals  in  turbid  waters.    Sewage  Indust.  Wastes.
29(6>:  695-711.

WARD, G.S.,  PARRISH,  P.R.  and KIGBY,  R.A.   (1981).  Early life  stage  toxicity
tests with a saltwater fish:  Effects of eight chemicals on survival,  growth, and
development  of  sheephead minnows  (Cyprinodon variegatus). J. Toxicol.  Environ.
Health.   8: 225-240.
                                     17-29

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                          18.  HEALTH EFFECTS SUMMARY

18.1.  EXISTING GUIDELINES AND STANDARDS
18.1.U  Air.  The Occupational Safety and Health Administration (OSHA) currently
limits occupational exposure to toluene to 200 ppm as an 8-hour,  time-weighted-
average   (TWA),   with   an   acceptable  ceiling   concentration  of   300 ppm
(HO CFR 1910.1000);  the acceptable maximum peak above the ceiling concentration
is 500 ppm  for a maximum  duration  of 10 minutes.  The  National Institute for
Occupational Safety and  Health (NIOSH, 1973) currently  recommends  an  exposure
limit of  100  ppm as an  8-hour TWA  with  a  ceiling of 200 ppm.   An  8-hour TWA
concentration  of 100 ppm  is also  recommended  by  the  American  Conference  of
Governmental  Industrial  Hygianists  (ACGIH,  I960)  as  a Threshold Limit  Value
(TLV) for toluene; the short-term (15 minute) exposure limit recommended by the
ACGIH is 150 ppm.  ACGIH (1980) has further noted that there may be significant
contribution to  the overall  exposure by the cutaneous route.
     Threshold limit values that have been established for occupational exposure
to toluene in other countries are listed as follows (Verschueren, 1977):
     USSR                       13 ppm  (50 mg/m3)          1972
     Czechoslavakia             52 ppm  (200 mg/m;;)         1969
     West Germany (BDR)       200 ppm  (750 mg/m^)         197'*
     East Germany (DDR)         52 ppm  (200 mg/m::)         1973
     Sweden                     98 ppm  (375 ing/nT3)         1975
     There are no standards  for general atmospheric pollution by toluene in the
United States, although  a  National  Ambient  Air  Quality Standard specifies that
nonmethane hydrocarbons  shall not exceed 0.24 ppm  (160  i/g/nr) as a maximum 3-
hour average  concentration (6 to 9  a.m.), more  than once per year (40 CFR 50).
Ambient  air  quality standards  have,  however,  been promulgated  for  toluene in
other   countries.     These   foreign  standards  are   summc.rized  as   follows
(Verschueren,  1977):
     Country                        Concentration             Averaging Time
     USSR
0.15 ppm (0.6 mg/nr)             20 min
0.15 ppm (0.6 mg/m3)             24 hr
     West Germany  (BSD)           15 ppm  (60 mg/m3)                30 min
                                  5 ppm  (20 mg/m3)                24 hr
                                      18-1

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     East Germany (DDR)           0.5 ppm (2.0 mg/rrl             30 min
                                  0.15 ppm (0.6 mg/nr)            24 hr
     Bulgaria                     0.15. ppm (0.6 mg/m^)            20 min
                                  0.15 ppm (0.6 mg/ra3)            24 hr
     Hungary                     13.3 ppm (50.0 mg/m^)            30 min
    ;s                              5.3 ppm (20.0 mg/m^)            24 hr
    •Hungary (protected areas)    0.16 ppm (0.6 mg/in^)            30 min
                                  0.16 ppm (0.6 mg/m3)            24 hr
     Yugoslavia                   0.16 ppm (0.6 mg/m^)
                                  0.16 ppm (0.6 nag/or)
20 min
24 hr
18.1.2.  Water.   The  Committee on Safe Drinking Water of the National Academy of
Sciences concluded in 1977 that toluene and its major metabolite, benzoic acid,
were relatively  nontoxic,  and that  there  was insufficient  toxicological  data
available to serve as a  basis  for  setting  a long-term ingestion standard (WAS,
1977).  It was recommended that studies be conducted to produce relevant informa-
tion.  Toluene has recently  been considered for a second time by a reorganized
Toxicology Subcommittee of the Safety  Drinking  Water  Committee of the National
Academy of Sciences  (U.S. EPA,  1980a),  but the  results of the deliberations of
thia group have net yet been made public.
     The U.S. EPA (1980a) has recently derived an ambient water criterion level
for toluene of 1^.3 mg/2..  This criterion is intended to protect humans against
the  toxic  effects of toluene  ingested through water  and  contaminated aquatic
organisms,  and is based on an Acceptable Daily Intake (ADI) calculated from the
maximum-no-effect dose reported in the Wolf et al. (1956) subchronic oral study
in rats-and an uncertainty factor of 1000.   The criterion level for toluene can
alternatively be  expressed  as 424 mg/X, If exposure is assumed  to  ue from the
consumption of fish and shellfish products  alone.
18,1.3.  Food.  Toluene has been approved by the Food and Drug Administration for
use as a component of articles intended for use  in  contact with foo^ (i.e., an
indirect food additive).   Articles that contain residues of toluene may be used
in   producing,   manufacturing,  packing,   processing,  preparing,   treating,
packaging,  transporting,  or holding food.   The use  of  toluene  in  the  food
industry is summarized as follows:

     Component of adhesives                               21 CFR 175.105
     Adjuvant substance in resinous and
       polymeric coatings for polyolefin films
       used as food contact surfaces                      21 CFR 175.320

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     Component of the uncoated or coatacl
       surfaces of paper and paper-board
       articles Intended for use with
       dry foods                                   "       21 CFR 176.1GO
     Used in the formulation of semirigid
       and ri^id acrylic and modified acrylic
       plasfj.c articles                                   21 CFR 177.1010
     Additive for cellophane (residue limit
       0.1*)                                              21 CFR 177.1200
     Additive for 1,4-cyclohexylene dimethy-'
       lene terephthalate and 1 , 4-eyclo-
       hexylene diroethylene isophthalate
       copolymer                                          21 CFR 172,1240
     Solvent for 4,4'-isopropylidenediphenol-
       epichlorohydrin resins with a minimum
       molecular weight of 10,000 (residue
       limit £1000 ppm in the finished resin)             21 CFR 177.1440
     Solvent for polysulfide polymer-polyepoxy
       resins                                             21 CFR 177.1650
     Solvent for poly(2,6-dimethyl-1,4-
       phenylene)oxide resins (residue limit
       0.2$ by weight)                                    21 CFH 177.2460
     Blowing agent adjuvant used in the manu-
       facture of foamed polystyrene (residue
       limit £0.35? by weight of finished
       framed polystyrene)                                21 CFR 178.3C"0
     Toluene has also been exempted from the requirement of a tolerance when it
is used as a solvent or cosolvent in pesticide formulations that are applied to
growing crops (40 CFR 180.1001).
18.2.  INHALATION EXPOSURES
     As detailed  in  Chapter 11 of  this  report,  many studies have reported the
effects on  humans  of inhalation exposures  to  toluene.   Because most  of these
studies  involved  relatively small  numbers of human subjects,  they  failed to
precisely  define  the  levels  or  durations of  the  exposures,  and/or  did  not
consider  the  potential role  of exposures  to  other toxic-ants.   None  of these
studies would be suitable for  human risk assessment if  taken individually.  In
combination, however,  they constitute a  considerable body  of human experience
and provide a relatively consistent pattern of dose-response relationships.
18.2.1.  Effects of Single Exposures.  The effects on humans of single exposures
to toluene for periods of up to 8 hours are relatively well documented.  Data on
both toluene glue sniffers (Press and Done,  1967a,  1967b; Wyae,  1973; Lewis and
Patterson,  1974;  Helliwell  and Murphy,  1979;  Hayden et al., 1977;  Oliver and
Watson, 1977; Barnes,  1979)  and  workers  accidentally exposed to high levels of
                                      18-3

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toluene (Lurie, 1919;  Browning,  1965;  Longley et  al.,  1967; Reisen et al., 1975)
indicate that  exposure to air  saturated or  nearly.saturated with  toluene can
cause a spectrum of effects, from lightheadedness to unconsciousness, in a very
short period of time.   Deaths attributed to the deliberate inhalation of toluene
have been  reported - in  at  least  21! cases  (Winek et  al.,  1968; Chiba,  1969;
Nomiyama and Nomiyama,  1978).   Although most of these reports  do  not provide
quantitative exposure  estimates,  glue sniffers  are  probably  exposed to nearly
saturated  air-vapor  mixtures of  about 30,000 ppm  toluene.    The  occupational
report of Longley et al.  (1967) indicated that a loss  of consciousness occurred
within minutes after  exposure  to  atmospheres estimated to  contain 10,000 ppm
toluene at waist level and 30,000  ppm  toluene at  floor  level.  The acute inhala-
tion toxicity  data  on experimental mammals,  summarized  in  Table 12-1, suggest
that exposure periods of  several  hours to  toluene levels greater than 4000 ppm
may  be  lethal.  Based  on the results of  longer term human  studies discussed
below, short exposures to concentrations of  up to 1500 ppm are not likely to be
lethal (Wilson, 19*43;  Ogata et al., 1970, see following discussion).  The single
report by  Gusev  (1965)  of effects on  EEC activity in  1 individuals exposed to
0.27 ppm for 6 minute intervals may be a subtle  indication of the perception of
toluene at  this  low level but does not  have any apparent  toxicologic signifi-
cance.
     For single  exposure periods  that approximate  a  normal  working  day  (7 to
8 hours),  von  Oettingen  et  al.  (19^2a, 19^2b) and Carpenter et al.  (19W pro-
vide relatively consistent information on sublethal dose-response relationships.
As summarized previously in Table  10-1, von Oettingen et al.  (.19*123,  19H2b) noted
a range of subjective complaints from  8 hour exposures  to toluene concentrations
ranging from 50 ppm (drowsiness)  to 800 ppm (severe fatigue, nausea,  incoordina-
tion, etc.,  with afte"  effects  lasting at  least several days).  Although the
terminology used by Carpenter et al. (19*1*0  is somewhat different from that used
by  von  Oettingen,  the  effects  noted   seem  comparable  over  the common exposure
range (200 to 800 ppm).   Although the consistency between  these two studies is
reassuring,  it  should  be  noted  that,  even combined,  both  studies  involve
exposures of only five individuals who were placed on multiple exposure/recovery
schedules.  The impact tnat such multiple exposures could potentially have on the
results cannot be determined.  Given the  small number of individuals involved in
the exposures  to toluene, an attempt  to generalize  for the human population a
detailed dose-response gradient comparable  to that presented in Table 11-1 does
                                      18-11

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not seem justifiable.  When these studies are considered along with the results
of Ogata and  coworkers  (1970)  and Gamberale and Hultengren (1972), however, it
seems reasonable to conclude that exposure periods  of 8 hours or less to toluene
concentrations below  100 ppm may result in mild subjective coaplaints (fatigue
or headache)  but  are  not  likely to induce  observable  effects.   Concentrations
above 100 ppm may cause impaired reaction time (200 ppm x 3 hours, Ogata et al.,
1970; 300 ppm x 20 minutes, Gamberale and Hultengren,  1972).  At concentrations
of 300 to 800 ppm and above, gross signs  of incoordination may be expected (von
Oettingen et al., 1942a, 19J42b;  Carpenter et al.,  19W.
     Accidental acute overexposure to  toluene may  be limited to some extent by
the organoleptic  or irritant properties of the compound.   Gusev (1965) reports
ranges of  maximum imperceptible concentrations and minimum perceptible concen-
trations of  0.35  to 0.79 ppm and 0.40  to 0.85 ppm, respectively.   May (1966)
reports  a  minimum perceptible  concentration  of 37 ppm.   The  reasons for this
discrepancy between the Russian  and American values are not apparent.  Although
the Russian  study entailed  a total of 30 subjects and 7M observations and the
American report involved 16 individuals  (number of observations not specified),
it  is  unlikely that  the difference  in the reported  detectable levels  is due
simply to sample size. In any event,  toluene appears to be detectable in the air
at levels below those causing impaired  coordination (i.e., >100 ppm).  In addi-
tion, Carpenter and coworkers C^M) reported that toluene caused mild throat and
eye irritation at 200 ppm and also caused lacrimation at 400 ppm.
     In summary, the estimated dose-response relationships for the acute effects
of single short-term  exposures  to toluene are presented below:

     10,000 to           :    Onset of  narcosis within a few minutes.  Longer
     30,000 ppm               exposures  may be lethal.
     >4,000 ppm          :    Would probably cause rapid impairment of reaction
                              time and  coordination.   Exposures  of  1  hour or
                              longer might  lead to narcosis and possibly death.
      1,500 ppm          :    Probably  not  lethal  for exposure periods of up to
                              8  hours.
        300 to 800 ppm   :    Gross  signs  of  incoordination  may  be  expected
                              during exposure periods up to 8 hours.
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        100 ppm          :    Lacrimation and irritation to the eyes and throat.
        100 to 300 ppm   :    Detectable signs of inooordination may be expected
                              during exposure periods up to 8 hours,
        200 ppm          :    Mild throat and eye irritation.
         50 to 100 ppm   :    Subjective  complaints  (fatigue or'headache)  but
                              probably no observable  impairment of reaction time
                              or coordination.
        >37 ppm          :    Probably perceptible to most humans.

From the above discussion,  it should be  evident  that  these  approximations  are
crude composites and contain several areas of uncertainty and overlap.
18.2.2.   Effects of  Intermittent Exposures  Over Prolonged  Periods.   Limited
information is  available on  the effects  of subchronic or  chronic  continuous
exposures to  toluene  on humans or experimental  animals.  Most of  the studies
either  involve  occupational  exposures  or  are  designed to  mimic  occupational
exposures.  Consequently, while the data  described below may be directly applic-
able to estimating effects from occupational exposures,  an additional element 01'
uncertainty must be  considered  in  any  attempt to estimate the  effects  of
continuous exposures that may occur from ambient  air.
     Wilson (19*13) provides the only acceptable data on  the effects of repeated
occupational exposures to toluene over a period of weeks (Section 11.1,1.2.).  In
this study,  the workers  were classified  into  three groups  by  the  levels  of
toluere to which they  were  exposed:   50  to 200 ppm,  200 to 500 ppm, and 500 to
1500 ppm.  The effects noted at the various  levels were  essentially the same as
those  seen  in single  exposures.  In  the  low  exposure group, the  reports of
headache and lassitude are  consistent with symptoms  noted  by von  Oettingen  and
coworkers (1942a, 19U2b) over the same range  of exposure. Although Wilson (19^3)
did not attribute these  effects  to  toluene exposure, his failure  to include an
unexposed  control  group makes   this  judgment  questionable  in  view  of  the
von Oettingen data.   In the  middle  and  high exposure  groups,  the  reports of
headache, nausea, and concentration-related impairment of coordination and reac-
tion time are also  consistent with the symptoms  reported by  von  Oettingen  and
coworkers  (19423,   1942b)  and Carpenter  and coworkers dg'lt) for  short-term
single exposures. The major discomforting feature of  the Wilson (19^3) report is
that it involved only 100 out of a total  of 1000 workers. It is unclear whether
the remaining 900 workers evidenced any symptoms  of  toluene exposure.
                                      18-6

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     The  only  other study that  reports  effects  of  repeated  exposures  to  toluene
fo»* relatively short periods of  time is that presented by Greenburg and coworkers
(1912).   In  this study,  repeated  occupational exposures to  toluene at levels of
100 to  1100  ppm  for periods  of  2  weeks  to  5  years  were  associated  with enlarged
livers  In 13 of 61 airplane painters.  This incidence  of liver-enlargement was
reported  to  be  3  times  that of a control group of  430 workers  not  exposed to
toluene.  Because Greenburg and coworkers (1942) were not able to  associate liver
enlargement  with  clinical  or  laboratory evid-ence of  disease,  because  the
painters  were also  exposed  to  significant quantities  of  other  volatile paint
•components (Table  11-9),  and because  the liver  effect has not  been corroborated
by  other investigators  (e.g.,  Parmeggiani  and Sassi,   1954;  Suhr,  1975),  the
hepatomegaly reported  by Greenburg should be given  relatively little weight in
risk assessment.
     Other reports of repeated occupational exposures to toluene  involvu  periods
of several years.   For mean  exposure  levels  above  200 ppm,  all of  the available
studies  except that of Suhr (1975)  report some evidence of neurologic  effects
(Capellini and Aleasio,  1971;  Panaeggiani and Sassi,  1954;  Munchinger, 1963;
Rouskova,  1975).
     The  Suhr (1975) study  involved  a  group of 100 pi-inters exposed to 200 to
400 ppm  toluene for over 10 years.   Compared to a group  of 100 non-exposed
individuals,  no  significant  differences were seen  in symptoms  of  CNS  depression
or sphallograph  tests, which are  designed  to measure muscular  coordination.  An
interpretation of  the  significance  of the Suhr  (1975) study  is confounded,
uowever,  by  several factors. As  discussed in Sections  11.1.1.2.  and  11.3-, the
limitations  of  this study  include  an undefined  control  group,  uncertainties
involving the time of reflex  reaction and sphallograph  testing  (i.e., blood
toluene  levels  may have declined significantly  if the workers  were examined
before or after the work  shifts),  and the use of an  apparently unvalidated device
(sphallograph)   for   the  detection   of   slight   disturbances   of  muscular
coordination.
     The  other  studies  that do  report effects at  equal  or  higher  levels of
exposure  can be  challenged for  various reasons.  The report of "nervous hyper-
excitability"  in  6  of  11 exposed to  200  to 800  ppm toluene  for  "many years"
(Parmeggiani and Sassi,  1954)  does  not seem  to  be characteristic of  toluene
intoxication.   This report is from the Italian literature,  however,  and a  full
text translation has not yet been made available for this review.   The Capellini
                                      18-7

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and Alessio (1971) study,  which  associated stupor,  nervousness,  and insomnia
with occupational  exposure to 250  (210  to  300) ppm toluene for several years,
involved  only  a  single  worker.    The  "organic  p.sychosy-ndrome"  diagnosed  by
Munchinger  (1963)  in  workers exposed  to  300  and  iJ30 pp.m toluene  for  18 and
12 years,  respectively,   is  supported  by  the  results  of Rorschach  tests and
Knoepfel's  13-Error  tests.   Because Munchinger  did not  use  a control group,
however* the utility of this  study  is limited.   The changes in EEC response to
photic stimulation that were reported by Rouskova (1975)  in workers exposed to
>250 ppm toluene for an average of 13.5 years'also involved exposure to unspeci-
fied  levels of  1,1,1-trichloroethane.   Thus,  the   interpretation  of  the dis-
crepancies between the study by Suhr (1975)  and  these other reports is problema-
tic.  Considering the relatively well documented CNS effects  of single exposures
to toluene  at  levels  above 200 ppm  (Section  18.1.1.)  and the  effects noted by
Wilson (19^3)  at comparable levels  for  much  shorter periods of time, it would
seem imprudent to accept the Suhr (1975) data as  a "no-obs-vrved-effect level" for
human risk assessment.
     An alternative approach could  be to use  the study  by Capellini and Alessio
(1971) in  which no CNS or  liver  effects were  noted in  a group  of 17 workers
occupationally  exposed to 125  (80 to 160) ppm toluene for "diverse  years."  In
addition to the problems  of small  sample size,  failure to precisely define the
duration of exposure, and  lack  of  a  control  group, the  use  of  this study is
compromised by reports of effects in two other groups of workers at lower levels
of toluene exposure.  Matsushita  and coworkers (1975) reported  impaired per-
formance in  neurological  and muscular  function tests  in  a group  of  38 feaale
shoemakers  who had  been  exposed to 15 to 200 ppm  toluene for  an average of
3 years and 4 months.  In  addition,  19 of 38  exposed women, compared to  3 of  16 in
the control group, complained  of dysmenorrhea.  The  second group of workers was
composed of 100 car painters who had  been occupat.ionally exposed to  an average of
30.6 ppm  toluene for an  average  of  14.8  years.   As reported  by  Hanuj.r,en and
coworkers  (1976) and Seppalainen and coworkers (1978),  the exposed  workers had a
greater incidence of CNS  symptoms and impaired performance on tests for intelli-
gence and memory, as well as for visual and  verbal ability.  Both of the studies
on this  group  of  worker  used  control groups or  approximately  100 unexposed
individuals.  The major problem with the reports of adverse effects  on the female
shoemakers  and male  car  painters  is that both  groups  were  exposed  to other
potentially toxic agents.  The female shoemakers were exposed to "alight" levels
                                      18-8

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of gasoline (Matsushita et al., 1975) and, as  detailed in Table  11-3, the raale
car painters were exposed to several  other organic solvents.
     Tl*e subchronic and chronic data on experimental mammals are of only limited
use in  helping to resolve the  uncertainties  in the human data.   Jenldns and
coworkers (1970), and CUT (1980)  report  no-observable-effect  levels  (NOELs) in
experimental mammals 1085 PF°> (8 hours per day, 5 days per week for 6  weeks) and
300 ppm  (6 hours  per day, 5 days  per week for  24 months),  respectively.   As
                                                          m~?
discussed above in  this  section,  a NOEL   of 1085  ppca  is  contradicted by human
experience, suggesting that humans are more sensitive  tnan experimental mammals
to toluene exposure.  Similarly, the  continuous-exposure  NOEL  of  H)7  ppm for 90
days in rats,  guinea pigs,  dogs,  and monkeys (Jenkins et al., 1970), and the
2 .year interaittant  exposure  NOELS of 30 ppm and 100  ppm in rats (CUT, 1980), do
not, by  themselves,  negate the concerns  with  neurological effects reported in
humans at lower levels.
18.3.  ORAL EXPOSURES
     Very little information  is available or.  the  acute,  subchronic,  or chronic
effects of toluene in experimental mammals.  As summarized in  Table 12-1, acute
oral LD^s in  adult  rats  range  from  5500 mg/kg to 7530 mg/kg.  Using the cubed
root of  the  body weight ratios for  interspecie?  conversion  (U.S. EPA, 1980b;
Freireieh et al., 1966;  Rail, 1969), an approximate lethal dose for humars can be
estimated  at  983 mg/kg  (5500 mg/kg  •  (70 kg   •  0.^ kg)  3).    The  conversion
factor, as used here, assumes that  humans are more sensitive than rats, which, as
discussed above, is  consistent  with the  available  data on inhalation exposure.
This estimate of the approximate lethal dose is also consistent with  the report
by  Francone  and Braier  (195*0  that leukemia  patients were  able to tolerate
cumulative  doses  of  up  to  130,000 ng of toluene  given over  a  3-week period
(approximately 88 mg/kg/day).
     The  only  subchronic  oral data  are reported  in the  study  by Wolf and
coworkers (1956), indicating  a NOEL in rats at 590  mg/kg/day, given  5 days per
week for 6 months.
18.H.  DERMAL EXPOSURES
     Studies on the  dermal toxicity of toluene are  inadequate  for quantitative
risk assessment.  Qualitatively, the little information  that  is available sug-
gests that moderate dermal contact with liquid toluene (i.e.,  exposure of human
forearm skin to toluene for  1 hour on C  successive  days) may  cause skin damage
but does not result  in overt  signs of toxicity  (Malten et  al., 1968).  Similarly,
                                      18-9

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the acute and subchronic data en toluene exposure  in experimental mammals do not
suggest  that  toluene is  a potent  toxicant  on dermal  contact.    A  method for
quantitatively using such data to estimate equivalent human dose-response rela-
tionships, however, has not been fully  formulated or validated.
     As discussed in Section  13.1.-,  exposure  to toluene vapor results Jn rela-
tively little densal absorption compared to absorption across the lungs,   **
18.5.  RESPONSES OF SPECIAL CONCERN
I8i5.1.   Carcinogenicity.   CUT (1960)  concluded  that  exposure  to 30, 100, or
300 ppm toluene  for  2*4  laonths did  not  produce an increased  incidence of neo-
plaatic,  proliferative,  inflammatory,  or  degenerative  lesions  in Fischer 3^
rats.   It  should be  noted,  however,  that   this  study  has been  considered
inadequate for carcinogenicity  evaluation  because the highest level tested was
not a  maximum  tolerated dose.  Also,  the  high spontaneous  incidence  (165) of
raononuclear cell leukemia in aging Fischer JkH male rats reported by Coleman and
coworkers (1977) suggests that this strain  may be  inappropriate for the study of
a chemical that might be myelotoxic.
     Other studies suggest  that  toluene is not carcinogenic when applied topi-
cally to the shaved skin of animals.   Toluene is  used extensively  as a solvent
for  lipophilic  chemicals  being  tested for  carcinogenic,  potential;  negative
control studies  employing  100J  ",oluene have  not elicited  carcinogenic effects.
Also, no evidence of a  promotion effect was r.cted when toluene was painted on the
skin of mice twice weekly for 20 weeks  following initiation with 7,12-dimethyl-
benz-[a]-anthracene (Frei and Stephens,  1968; Frei cind Kingsley,  1968).
     The above data are not adequate for assessing the potential carcinogenicity
of toluene with great assurance and they cannot be used for supporting carcino-
genicity as a valid biologic endpoint in quantitative risk assessment.
18.5.2.   Mutagenicity.   Toluene  has yielded negative results in  a  battery of
microbial, mammalian cell, and  whole  organism test  systems as indicated in the
following:

          Differential Toxicity/DNA Repair Assays
               Escherichia ccli
               Salmonella typhimurium
          Reverse Mutation Testing
               Salmonella typhimuriun  (Ames test)
               Escherichia coli WP2 assay
               Saccharomyces cerevisiae D7
                                     18-10

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          Mitotic Gene Conversion/Crossing Over
               Saccharomyoes cerevisiae D4, D7
          Thymidine Kinase Assay
               L5178Y mouse iymphoma cells
          Micronucleus Test
               mouse
          Dominant Lethal Assay
               mouse
     Assays for sister-chronsatid exchange (SCE) and cytogenetic effects in human
and animal systems  have  provided inconsistent results.  In  vitro studies' have
shown that toluene  treatment  did not  alter  SCE  frequencies  in cultured Chinese
hamster ovary cells  (Evans  and  Mitchell,  1980),   and  that SCEs and  chromosome
aberrations  were  not induced in cultured human lymphocytes  (Gerner-Smidt and
Friedrich,   1978).    Increased  frequencies  of  SCEs  and/or  aberrations in
lymphocytes  from  workers  who were  chronically  exposed to  similar  levels of
toluene  have,  however,  been  reported by some  investigators (100 to  200 ppm,
Funes-Craviota et al., 1977; 200 to 300 ppm, Eauchingsr et al.,  1982), but not by
others (200 to 400 ppm,  Forni  et al.,  1971; 7 to 112 pp-n, Maki-Paakkanen et al.,
1980).  In the  Russian  literature, chromosome aberrations were reported in the
bcne marrow  cells of  rats  that  were exposed subcutaneously  (Dobrokhotov,  1972;
Lyapkalo, 1973) and via  inhalation (Dobrokhotov and  Enikeev,  1977) to  toluene,
but these findings were not corroborated in a Litton Bionetics, Inc. (1978)  study
with rats  following intraperitoneal injection.  Differences in doses  employed
and experimental design (e.g., numbers of cells scored) may account (at  least in
part) for the  conflicting results, but it  should be  noted that it is  probable
that part  of the exposure in  the Funes-Craviota  et  al. (1977)  study was to
benzene-contaminated  toluene, and that the purity of the toluene used in the
Russian studies was not stated.
18.5.3.  Teratogenicity.  Toluene was  reported in a recent abstract from NIEHS to
induce cleft  palates at a level of 1.0 m£/kg  (approximately  866 mg/kg) following
oral exposure to  mice on days 6 to 15 of gestation (Nawrot  and Staples,  1979).
This effect reportedly did not appear to be due merely to a  general retardation
in growth rate.   Levels  of 0.3  and 0.5 mS,/kg  (approximately  260 and  433 mg/kg)
toluene had no teratogenic effect, but the number  of  mice exposed  and nunber of
fetuses  examined  were  not  stated.   Nawrot  and  Staples (1979)  also  noted a
significant increase in embryonic lethality  at all dose levels and  a significant
                                     18-11

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reduction in  fetal  weight at  the. two higher dose  levels.   No  frank  signs of
maternal toxicity  were seen at  any dose level; however, at  the highest dose,
decreased maternal weight gain was reported in mice exposed on days 12 to 15 of
gestation.  A complete copy of  this report has not been made  available for review
but has been submitted for publication.
     Three other studies have concluded  that toluene is not teratogenic in mice
(Hudak and Ungvary,  1978)  or rats  (Hudak and Ungvary,  1978;  Litton Bionetic .,
1978; Tatrai  et al., 1979)  following  inhalation exposure.   Hudak  and Ungvary
(1978) and Tatrai et al.  (1979)  have noted,  however,  an increased incidence of
skeletal anomalies and signs of  retarded skeletal.development in the rats that
were not considered malformations as such.  Embryotoxicity was also indicated by
low fetal weights in  mice and some rats  (Hudak and Ungvary,  1978).  At the high
exposure levels in  the study  by Hudak and Ungvary  (1978),  increased  maternal
mortality  was  noted  in  rats  (399  ppm,  2*4  hours/day,  days  1  to 8)  and  mice
(399 ppm, 24 hours/day, days 6  to 13).  No increased maternal mortality was noted
by  either  Hudak and  Ungvary (1978) or  Tatrai et  al.  (1979)  at lower exposure
levels  in  rats (266 ppm,  8 hours/day,  days 1   to  21;  266 ppm,  24 hours/day,
days 7 to  1*0  or mice (133  ppm,  24  hours/day, days  6  to 13).   In the study by
Litton Bionetics, Inc.  (1978), no signs  of maternal toxicity were noted in rats
exposed to 100  or 400 ppm, 6 hours/day,  on days  6 to 15 of gestation.
     The extrapolation of these results to  define potential human risk  is an
uncertain process.   The  dose that produced cleft palates in mice on oral expo-
sure, 866 mg/kg, is  only slightly  higher than the NOEL in rats, 590 mg/kg/day.
     Although  inhalation exposure  to  toluene has  not been shown  to  be tera-
togenic, embryotoxicity is an endpoint  of concern.  The  effects  noted in rats and
mice  at  the high  exposure level  (400 ppm)  in  the study by  Hudak  and Ungvary
(1978) may be of limited use in  human  risk assessment because of the occurrence
of  maternal  mortality.   The lowest effect  level  not  associated with maternal
mortality was  133  Ppm,  24 hours/day, on days 6  to 13, which  caused  low fetal
weights in mice.  No  fetal effects were  noted in the study by Litton Bionetics,
Inc. (1978), however, when rats were exposed  to 100  ppro or 400 ppm, 6 hours/day,
on days 6 to 15 of gestation, or  in the Tatrai et al. (1979)  study when  rats were
continuously exposed  to  266  ppm  toluene  on days  7  to 14.  As  is  the  case with
oral exposure  studies,  a quantitative approach  for  using  this type of data in
human risk assessment has not  been validated.
                                      18-12

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18. 6-.  REFERENCES

ACGIH  (AMERICAN  CONFERENCE OF  GOVERNMENTAL INDUSTRIAL  HYGIENISTS).    (1980).
Documentation of the Threshold Limit Values -for Substances in Workroom Air,  tth
ed.f Cincinnati, OH-  p.
BARNES, C.E.  (1979).  Solvent abns*:.  A review.  Int. J_. Addict.  _VJ:  1-26.

BAUCHINGER, M., SCHMID, E., DRESP, J., KOLIN-GERRESHEIH, J., HAUF.  R.  and SUHR,
E.   (1982).   Chromosome changes  in lymphocytes  after occupational exposure  to
toluene.  Hutat. Res.
BROWNING,  E.    (1965).   Toxicity and Metabolism  of Industrial Solvents.   New
York: Elsevier Publishing Co., pp. 66-76.

CAPELLINI, A. and ALESSIO,  L.   (1971).  The urinary excretion of hippuric acid in
workers exposed to toluene.  Med. Lavoro.  62: 196-201.  (In Ital.).

CARPENTER, C.P., SHAFFER, C.B., WEIL, C.S. and SMYTH, H.F.,  JR.  (WO.  Studies
on the inhalation of  1,3-butadiene; with a comparison of its narcotic effect with
benzol,  toluol,  and styrene,  and a  note on the elimination of styrene  by the
human.  £. Ind.  Hyg. Toxicol.  26: 69-78.

CHEMICAL INDUSTRY INSTITUTE OF TOXICOLOGY (CUT).   (1980).   A  twenty-four month
inhalation toxicology study in Fischer-S^I rats exposed to atmospheric toluene.
Executive  Summary   and  Data  Tables.    Conducted   by   Industrial   Sio-Test
Laboratories,  Inc.  and  Experimental  Pathology Laboratories, Inc., Raleigh,  NC
for CUT, Research Triangle Park, NC.  October 15,  1980.

CHIBA,  R.    (1969).    Sudden  death  from  thinner.   Nichidai  Igaku  Zasshi.
2!8: 982-998.  Taken from: Chem. Abst.  72: 64867g,  1969-

COLEMAN, G.L., BARTHOLD, S.W.,  OSBALDISTON,  G.W.,  FOSTER,  S.J. and JONES,  A.M.
(1977).   Pathological  changes during aging  in barrier-reared Fischer 3M  male
rats.  J. Gerontology.  _3_2: 258-278.
                                     18-13

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       '     ' V"Bt  (1972).  Ine mutagenic influence of benzene and toluene under
experimental conditions.  Gig. sanit.  J7:  36-39.  (In Russian; evaluation based
on an English translation provided by the U.S. EPA).

DOBROXHOTOV, v.B.  and  EfilKEEV , M.I.   (1975).   Hutagenic effect of  benzene,
toluene,  and  a mixture of  these hydrocarbons in a  chronic experiment.   Gig.
sSB-y-*   !'• 32-3*4.    (in  Russian;  evaluation  based  on an  English  translation
provided  by the U.S. EPA).

EVANS, E.L., and MITCHELL, A .D .  (1980).  An Evaluation of  the Effects of Toluene
or.  Sister  Chroma t id  Exchange  Frequencies -in Cultured  Chinese  Hamster  Ovary
Cells.  Prepared by SRI International, Menlo Park, CA , under Contract No.  68-02-
     for  the U.S.  Environmental Protection Agency,  Research Triangle  Park,  NC.
FORHI, A., E.  PACIFICO and A. LIMCWTA.   (1971).  Chromosome studies in workers
exposed  to benzene  or toluene or both.  Arch. Environ. Hearth.  22:  373-378.

FRANCONE, M.P. and BRAIER , L.  (195*0.  The basis for the substitution of benzene
by the higher  homologues in  industry.  Med. Lavoro.  4j>:  29-32.  (In Ital.)".

FREI,  J.V.  and STEPHENS,  P.   (1968).   The  correlation  of promotion  of  tumor
growth and  of induction of  hyperplasia  in epidermal  two-stage carcinogenesis .
Brit. £. Cancer.  22: 83-92.

FREI,  J.V.  and  KINGSLEY,  W.F.    (1968).   Observations  on chemically  induced
regressing tumors of mouse epidermis.  «J. Natl. Cancer Inst.  JH:  1307-1313-

FREIREICH,  E.J., GEHAN ,  E.A.,  RALL,  D.P.,  SCHMIDT,  L.H,  and SKIPPER,  H.E.
(1966).  Quantitative comparison of toxicity  of anticancer agents in mouse,  rat,
hamster, dog,  monkey, and  man.  Cancer Chemother. P&p.  ^0:  219.

FUNES-CRAVIOTA , F., et al.   (1977).  Chromosome aberrations and sister-chroaatid
exchange in  workers in chemical laboratories and a rotoprinting factory and  in
children of women laboratory workers.  Lancet.  2:  322.
                                      18-11

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                               I
GAMBERALE,  F.  and  HULTENGREN,  M.   (1972).    Toluene exposure.   114 Phychop-
hysiological functions.  Worlc Environ. Health.  3(3)1; 131-139.

GERNER-SMIDt,  P.  and FRIEDRICH,  0.   (1978).  The  mutagenic effect  of benzene,
toluene, and xylene studied by the SCE technique.  Mutat. Reg.  58(2-3);  313.

GREEMBURG, L., MAYERS, M.R,., HEIMANN, H.  and MOSKOWITZ, S.  (19'I2).  The effects
of exposure to toluene in  industry.  J_. Amer. Hed. Aaaop.  118;  573-578.

GUSEV,  I.S.   (1965).  Reflective  effects  of microconcentrationa  of benzene,
tolusne, xylene,  arid their comparative assessment.   Hjrg. Sanit.   _3_0: 331-335.
(Russian report published  in English).

HANNINEN,  H.,  ESKELINEN,  L., HUSMAN, K. and  NURMIMEN,  M.   (1976).   Behavioral
effects of long-term exposure to a mixture of organic solvents.   Scand. J.  Work
Environ. Health.  2(*Q: 2UO-255.
    j
HAIDEK, J.W.,  PETERSON, R.G. and BRUCKNER, J.V..  (1977).  Toxicology of toluene
(raethylbenzene):  Review of current literature.  Clin. Toxicol.  11(5): 5^9-559.

HELLIWErX,  M.  and   MURPHY,  M.    (1979).   Drug-induced neurological  disease.
(letter).   Brit.  Med. J.   1(6173):  1283-1284.

HUDAK, A.  and  UNGVARY, G.  (1978).   Embryotoxic effects of benzene and its methyl
derivatives:   Toluene and  xylene.  Toxicology.  11: 55.

JENKINS, L.J., JR., JONES, R.A. and SIEGEL,  J.   (1970).  Long-term inhalation
screening  studies  of benzene,  toluene,  o-xylene,  and cumene  on experimental
animals.   Toxicol.   Appl.  Pharmacol.   16: 818-823-

LEWIS,  P.W.,   and PATTERSON, D.W.    (WO.   Acute and chronic  effects  of the
voluntary  inhalation of certain  commercial  volatile solvents  by Juveniles.  J.
Orug issues.   *«(2)_; 162-175.
                                      18-15

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 I,

LITTON 3IONETICS,  INC.   (1978).   Teratology Study in  Rats.   Toluene.   Final
Report.   Submitted  to the American  Petroleum  Institute,  Washington, D.C.  in
January, 1978.  LBI Project No. 20698-1.  Litton Bionetics, Inc., Kensington,  MD.
17 P.

LONGLEY, E.O., JONES,  A.T., WELCH, R. and LOMAEV,  0.   (-1967).  Two acute toluene
episodes in merchant ships.  Arch. Environ. Health.  jUl: 181-187.

LURIE, J.B.   (1919).   Acute toluene poisoning.  £. Africa Hed. J_.  2_3_: 233-236.

LYAPKALO, A.A.  (1973). Genetic activity of benzene and  toluene.  Gig. Tr. Prof.
Azbol.   17; 2U-28.   (In  Russian with English summary;  evaluation  based  on an
English translation provided by  the U.S. EPA).

MAKI-PAAKKANEN,  J., et al.    (1980).   Toluene exposed  workers  and chromosome
aberrations.  J. Toxico]. Environ. Health.  ^: 775.

MALTEN,  K.E., SPRUIT, D.  and  DEKEIZER,  M.J.M.   (1968).  Horny layer injury by
solvents.   Berufsdermatosen.   16;'  135-147.

MATSUSHITA,  T., et al.   (1975).   Hematological  and neuro-muscular response of
workers  exposed to low concentrations of toluene  vapor.  Ind. Health.  13: 115.

MAY,  J.   (1966).    Odor thresholds of solvents for assessment of solvents odors
 in  the air.   Straub.  26(9): 31-38-

MUNCHINGER,  R.   (1963).  Der uachweis central nervoser storungen  bei losungsmitt
 el  ,.,-onierten Arbeitern.  Excerpta Medea Series, Madrid;  16-21. 2(62): 687-689,

NAS (National Academy of Sciences).    (1977).  Drinking Water and Health.  Safe
Drinking Water  Committee, Advisory  Center  on   Toxicology,  Assembly of-Life
Sciences,  National Research Council, National Academy  of Sciences, Washington,
DC.,  p.  939.  Available from Printing and Publishing Office, National  Academy of
Sciences,  2101  Constitution Ave.,  Washington, D.C.   20*418.
                                      18-16

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NAWBOT, P.S. and STAPLES, R,E.  (1979),'  Embryo-fetal toxicity and teratogenicity
of benzene and  toluene  in the mouse.  Teratology.  J9: 41A.  (Abstract).

NIOSH (National Institute for Occupational Safety and Health).   (1973).  Criteria
for.a  Recommended Standard.  Occupational  Exposure to Toluene.   Final Report.
Contract  No. HSM-99-72-118.   Available throng  NTIS,  NTI3  No. PB-222-?19/8-
108  pp.

NOMIYAMA, K. and H. NOMIUMA.  (1978).  Three fatal cases of thinner sniffing,
and  experimental  exposure to toluene  in humans and animals.   Int. Arch. Occug.
Environ.  Health.  4J;  55-64.

OGATA,  M.,  TOMOKUNI, K., TAKATSUKA, Y.  (1970).  Urinary excretion of hippuric
acid and m- or p-methylh.ippurie acid in the urine of persons exposed to vapors of
toluene and m-  or  p-xylene  as  a  test   of  exposure.    Brit.  J.  Ind.  Ned.
27(1);  43-50.

OLIVER, J.S. and WATSON, J.M.  (1977).  Abuse of solvents "for  hicks":  A review
of 50  cases.  Lancet.  1(6002): 84-86.

PARMEGGIANI, L. and SASSI, Co.  (1954).  Occupational ri*k of toluene:  Environ-
mental studies  and  clinical  investigations  of  chronic  intoxication.    Mod.
Laboro. _45: 574-583.

PRESS,  E.  and DONE, A.K.   (1967a).  Solvent sniffing.   Physiologic effects and
community  control measures for intoxication from the intentional inhalacion of
organic solvents,  r. Pediatrics.   ^9:  451.

PRESS,  E.  and DONE, A.K.  (1967b).  Solvent sniffing.  Physiologic effects, and
community  control measures for intoxication from the intentional inhalation of
organic solvents.  II.  Pediatrics. .39:  611.

RALL   D.P.    (1969).    Difficulties  in extrapolating the results  of toxicity
studies in laboratory animals to man.   Environ. Res.  2:  360-367.
                                      1C-17

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HEISIN, E., TEICHER, A., JAFFE, R. and ELIANHOU, H.E.  (1975). Myoglobinuria and
renal failure in toluene poisoning.  Brit. ±.  Indust. Hed.   32(2);  163-164.

ROUSKOVA, V.  (1975).  Photic stimulation  in early diagnosis of the effects of
some harmful  industrial  substances on  the central nervous system.  Int. Arch.
Arbeitsmed.  314(4); 283-299.

SEPPALAINEN,  A.M.,  HUSMAN,  1C. and MARTENSON,  C.   (1978).  Neurophysiological
effects  of  long-term exposure to a mixture of  organic solvents.  Scand. jr. Work
Environ. Health.  _4(4): 304-314.  taken from:   Chem. Abst.   90:  156383*,  1979.

SUHR, E.   (1975).  Comparative Invest!vation of the State of Health of Gravure
Printers Exposed  to Toluene.  Gesellschaft zur Forderung des Tiefdrucks E.V.,
Weisbaden,  Federal Republic of Germany.  92 pp.

TATRAI,  E., HUDAK,  A.  and UNGVARY, G .<   (1979).  Simultaneous effect on the rat
liver  of benzene,  toluene,  zylene, and CCL4.   Aeta.  Physiol.  Acad. Sci. Hung.
53(2); 261.

VERSCHUEREN,  K.   (1977).   Handbook of  Environmental Data  en Organic Chemicals.
New York,  NY:  Van Nostrand Reinhold Co., pp.  592-596.

VON OETTINGEN,  W.F.,  MEAL, P.A. and DONAHUE,  D.D.  (1942a).  The  toxicity and
potential   dangers  of  toluene—Preliminary report.    «J.  Amer.   Med.  Assoc.
 118; 579-584.

VON   OETTINGEN,   W.F.,   NEAL,   P.A.,   DONAHUE,  D.D.,   SVIRBELY,   J.L.,
BAERNSTEIN, H.D., MONACO,  A.R., VALAER,  P.J. and  MITCHELL, J.L.  (1942b).  The
Toxicity and Potential Dangers of Toluene, with Special Reference to its Maximal
Permissible Concentration.   U.S.  Public  Health  Service.   Pub.  Health Bull.
No. 279, 50 pp.

U.S. EPA (U.S. Environmental Protection Agency).  (1980a).  Ambient Water Quality
Criteria for Toluene.   Publication No. EPA 440/5-80-075.   U.S. Environmental
Protection Agency, Washington, D.C.
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                                   TECHNICAL REPORT DATA
                            (rleaie read Jnururiiom on the reverse htfori- completing!
1. REPORT NO.
    EPA-600/8-82-008 F
             3. RECIPIENT'S ACCESSION NO.

               PB  84-100056	
    '-? AND SUBTITLE

   HEALTH ASSESSMENT  DOCUMENT FOR
   TOLUENE—Final Report
             6. REPORT DATE
               August  1983
             6. PERFORMING ORGANIZATION CODE
 '. AOTMORIS)
9. PERFORMING ORGANIZATION NAME AND ADDRE~SS
    Syracuse Research  Corporation
    Center for Chemical  Hazard Assessment
    Merrill Lane
    Syracuse, NY  13210
                                                            . PERFORMING ORGANIZATION REPORT NO
             10. PROGRAM EuEMENT NO.
             IV CONTRACT/GRANT NO.

               68-02-3277
12. SPONSORING AGENCY NAME AND ADDRESS
    U.S. Environmental  Protection Agency
    Environmental  Criteria and Assessment Office
    Office of Research and DeveloDment
    Research Triangle Park,  NC 27711
             13. TYPE OF REPORT AND PERIOD COVERED
              4. SPONSORING AGENCY CODE
               EPA/600/22
IS-SUPPLEMENTARY NOTES
           j^e hg-jj.^ effect  of primary concern with regard to exposures of humans  to
 toluene is dysfunction of  the central nervous system (CNS). Occupational exposures in
 the range of 200 to 1,500  ppm have elicited dose-related CNS alterations. Although
 myelotoxicity was previously attributed to toluene, recent evidence indicated  that
 toluene is not toxic to  the  blood or bone marrow; myelfctoxic effects are considered to
 have been the result of  concurrent exposure to benzene.
           Available evidence is inadequate for assessing the carcinogenic potential of
 toluene. Although a 24-month inhalation exposure of rats to 300 ppm did not  produce any
 positive carcinogenic effects, various design deficiencies precluded the usefulness of
 this study in assessing  carcinogenic potential.
           Toluene has been shown to be non-mutagenic in a battery of microbial,  mamma-
 lian cell, and whole organism test systems. Animal exposure studies suggest  that
 toluene has low teratogenic  potential. However, tmbryototcicity has been shown  to be an
 endpoint of concern. The reproductive effects of toluene is a category recommended for
 additional research.
           Based on available exposure estimates, the only group at possible  hign risk
 are workers exposed at or  near the Threshold Limit Value  (100 ppm).
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