BENZENE
                                               r
                                               c



                                              s!
Agency for Toxic Substances and Disease Registry

U.S. Public Health Service

-------
                                                       ATSDR/TP-88/03
           TOXICOLOGICAL PROFILE FOR
                     BENZENE
              Date Published — May 1989
                     Prepared by:

            Oak Ridge National Laboratory
   under DOE Interagency Agreement No. 18S7-B026-A1

                         for

Agency for Toxic Substances and Disease Registry (ATSDR)
               U.S. Public Health Service

                 in collaboration with

      U.S. Environmental Protection Agency (EPA)
       Technical editing/document preparation by:

            Oak Ridge National Laboratory
                        under
     DOE Interagency Agreement No. 1857-B026-A1

-------
                          DISCLAIMER

Mention of company name or product does not constitute endorsement by
the Agency for Toxic Substances and Disease Registry.

-------
                                FOREWORD

     The Superfund Amendments and Reauthorization Act of 1986 (Public
Law 99-499) extended and amended the Comprehensive Environmental
Response, Compensation, and Liability Act of 1980 (CERCLA or Superfund).
This public law (also known as SARA) directed the Agency for Toxic
Substances and Disease Registry (ATSDR) to prepare toxicological
profiles for hazardous substances which are most commonly found at
facilities on the CERCLA National Priorities List and which pose the
most significant potential threat to human health, as determined by
ATSDR and the Environmental Protection Agency (EPA).  The list of the 100
most significant hazardous substances was published in the Federal
Register on April 17, 1987.
     Section 110 (3) of SARA directs the Administrator of ATSDR to
prepare a toxicological profile for each substance on the list. Each
profile must include the following content:

     "(A)  An examination, summary, and interpretation of available
     toxicological information and epidemiologic evaluations on a
     hazardous substance in order to ascertain the levels of significant
     human exposure for the substance and the associated acute,
     subacute, and chronic health effects.
     (B)  A determination of whether adequate information on the health
     effects of each substance is available or in the process of
     development to determine levels of exposure which present a
     significant risk to human health of acute, subacute, and chronic
     health effects.

     (C)  Where appropriate, an identification of toxicological testing
     needed to identify the types or levels of exposure that may present
     significant risk of adverse health effects in humans."

     This toxicological profile is prepared in accordance with
guidelines developed by ATSDR and EPA. The guidelines were published in
the Federal Register on April 17, 1987. Each profile will be revised and
republished as necessary, but no less often than every three years, as
required by SARA.
     The ATSDR toxicological profile is intended to characterize
succinctly the toxicological and health effects information for the
hazardous substance being described. Each profile identifies and reviews
the key literature that describes a hazardous substance's toxicological
properties. Other literature is presented but described in less detail
than the key studies. The profile is not intended to be an exhaustive
document; however, more comprehensive sources of specialty information
are referenced.
                                                                     iii

-------
 Foreword


      Each toxicological profile begins with a public health statement,
 which describes  in nontechnical language a substance's relevant
 toxicological properties. Following the statement Is material that
 presents levels  of significant human exposure and, where known,
 significant health effects. The adequacy of information to determine a
 substance's health effects is described in a health effects summary.
 Research gaps in toxicologic and health effects information are
 described in the profile. Research gaps that are of significance to
 protection of public health will be identified by ATSDR,  the National
 Toxicology Program of the Public Health Service, and EPA.  The focus of
 the profiles is  on health and toxicological information;  therefore, we
 have included this information in the front of the document.-

      The principal audiences for the toxicological profiles are health
 professionals at the federal,  state, and local levels,  interested
 private sector organizations and groups,  and members of the public. We
 plan to revise these documents in response to public comments and as
 additional data become available;  therefore,  we encourage  comment that
 will make the toxicological profile series of the greatest use.

      This profile reflects our assessment of all relevant  toxicological
 testing and information that has been peer reviewed.  It has been
 reviewed by scientists from ATSDR,  EPA,  the Centers  for Disease Control,
 and the National Toxicology Program. It  has also been reviewed by a
 panel of nongovernment peer reviewers  and was made available for public
 review.  Final responsibility for the contents and views expressed in
 this toxicological  profile resides  with  ATSDR.
                                     James 0. Mason, M.D.,  Dr.  P.H.
                                     Assistant  Surgeon General
                                     Administrator, ATSDR
iv

-------
     -«« 	  iii
LIST OF FIGURES 	   ix
LIST OF TABLES 	   xi

 1.   PUBLIC HEALTH STATEMENT 	    1
     1.1  WHAT IS BENZENE? 	    1
     1.2  HOW MIGHT I BE EXPOSED TO BENZENE? 	    1
     1.3  HOW DOES BENZENE GET INTO MY BODY? 	    1
     1.4  HOW CAN BENZENE AFFECT MY HEALTH? 	    2
          1.4.1  Brief Exposure at High Levels 	    2
          1.4.2  Long-Term Exposures at Various Levels 	    2
     1.5  IS THERE A MEDICAL TEST TO DETERMINE
          IF I HAVE BEEN EXPOSED TO BENZENE? 	    2
     1.6  WHAT LEVELS OF EXPOSURE HAVE RESULTED IN HARMFUL
          HEALTH EFFECTS? 	    3
     1.7  WHAT RECOMMENDATIONS HAS THE FEDERAL GOVERNMENT
          MADE TO PROTECT HUMAN HEALTH? 	    3

 2.   HEALTH EFFECTS SUMMARY 	    7
     2.1  INTRODUCTION 	    7
     2.2  LEVELS OF SIGNIFICANT EXPOSURE 	    8
          2.2.1  Key Studies and Graphical Presentations 	    8
                 2.2.1.1  Lethality	    8
                 2.2.1.2  Systemic/target organ toxicity 	   13
                 2.2.1.3  Developmental toxicity 	   15
                 2.2.1.4  Reproductive toxicity 	   16
                 2.2.1.5  Careinogenieity 	   16
          2.2.2  Biological Monitoring as a Measure of
                 Exposure and Effects 	   17
                 2.2.2.1  Monitoring of exposure 	   17
                 2.2.2.2  Monitoring of effects 	   20
          2.2.3  Environmental Levels as Indicators of
                 Exposure and Effects 	   20
                 2.2.3.1  Levels found in the environment 	   20
                 2.2.3.2  Human exposure potential 	   21
     2.3  ADEQUACY OF DATABASE 	   21
          2.3.1  Introduction 	   21
          2.3.2  Health Effect End Points 	   22
                 2.3.2.1  Introduction and graphic summary 	   22
                 2.3.2.2  Descriptions of highlights of graphs 	   25
                 2.3.2.3  Summary of relevant ongoing research 	   25

-------
 Contents

           2.3.3  Other Information Needed for Human
                  Health Assessment 	         28
                  2.3.3.1  Pharmacokinetics and mechanisms  of action ..   28
                  2.3.3.2  Monitoring of human biological samples  	   29
                  2.3.3.3  Environmental considerations  	   30
  3.   CHEMICAL AND PHYSICAL INFORMATION 	                 31
      3.1  CHEMICAL IDENTITY 	' [ ' '	   31
      3 .2  PHYSICAL AND CHEMICAL PROPERTIES 	'.'.'.'.'.'.'.'.'.'.'.'.'.'.'.  '. . .   31
  4.   TOXICOLOGICAL DATA 	                           35
      4.1  OVERVIEW 	' ' .' ' ]	   35
      4.2  TOXICOKINETICS 	'.'.'.'.'.'.'.'.'.'.'.'.'."".'.   35
           4.2.1  Overview 	   35
           4.2.2  Absorption 	    36
                  4.2.2.1  Inhalation 	             36
                  4.2.2.2  Oral 	'.'.'.'.'.'..'.   37
                  4.2.2.3  Dermal 	\\   37
           4.2.3  Distribution 	'.'.'.'.'.'.   38
                  4.2.3.1  Inhalation 	   38
                  4.2.3.2  Other routes  of  exposure  	               39
           4.2.4  Metabolism 	"j   39
           4.2.5  Excretion 	   43
                  4.2.5.1  Inhalation 	'.'.'.'.'.   43
                  4.2.5.2  Oral 	'.'.'.'.'.   44
                  4.2.5.3  Dermal 	               44
      4. 3   TOXICITY 	'.'.'.'.'.'.'.'.'.'.'.'.   44
           4.3.1  Lethality and Decreased Longevity  	   44
                  4.3.1.1  Overview 	   44
                  4.3.1.2   Inhalation 	                 46
                  4.3.1.3   Oral 	    '   47
                  4.3.1.4  Dermal 	'.'.'.'.'.'.   47
           4.3.2  Systemic/Target Organ  Toxicity  	   47
                  4.3.2.1   Hematotoxicity 	   47
                  4.3.2.2   Immunotoxicity 	   54
                  4.3.2.3   Neurotoxicity 	   57
                  4.3.2.4   Dermal toxicity  	   58
                  4.3.2.5   Ocular toxicity  	   58
           4.3.3  Developmental Toxicity 	   59
                  4.3.3.1   Overview 	   59
                  4.3.3.2   Inhalation	   59
                  4.3.3.3   Oral 	|.   63
                  4.3.3.4   Dermal 	   63
                  4.3.3.5   Injection  	   63
                  4.3.3.6   General  discussion  	   64
           4.3.4  Reproductive  Toxicity  	   64
                  4.3.4.1   Overview 	   64
                  4.3.4.2   Inhalation  	   65
                  4.3.4.3   Oral  	   65
                  4.3.4.4   Dermal  	   65
                  4.3.4.5   Injection  	   66
                  4.3.4.6  General  discussion  	   66
vi

-------
                                                                  Concents

         4.3.5  Genotoxlcity 	   66
                4.3.5.1  Overview 	   66
                4.3.5.2  Human 	   66
                4.3.5.3  Animal 	   67
                4.3.5.4  In vitro 	   70
                4.3.5.5  General discussion 	   70
         4.3.6  Carcinogenicity 	   73
                4.3.6.1  Overview 	   73
                4.3.6.2  Inhalation	   73
                4.3.6.3  Oral 	   85
                4.3.6.4  Dermal 	   91
                4.3.6.5  General discussion 	   91

5.   MANUFACTURE, IMPORT, USE, AND DISPOSAL 	   95
    5.1  OVERVIEW 	".	    95
    5.2  PRODUCTION 	    95
    5.3  IMPORT 	   96
    5.4  USE 	   96
    5.5  DISPOSAL 	   96

6.   ENVIRONMENTAL FATE 	   99
    6.1  OVERVIEW 	   99
    6.2  RELEASES TO THE ENVIRONMENT 	   99
    6.3  ENVIRONMENTAL FATE 	  102
         6.3.1  Transport 	  102
         6.3.2  Transformation and Degradation 	  103
                6.3.2.1  Chemical degradation 	  103
                6.3.2.2  Biodegradation 	  104
7.   POTENTIAL FOR HUMAN EXPOSURE 	  105
    7 .1  OVERVIEW 	  105
    7.2  LEVELS MONITORED OR ESTIMATED IN THE ENVIRONMENT 	  105
         7.2.1  Air 	  105
         7.2.2  Water 	  108
         7.2.3  Soil 	  108
         7.2.4  Other 	  108
    7. 3  OCCUPATIONAL EXPOSURES 	  110
    7.4  POPULATIONS AT HIGH RISK 	  110

8.   ANALYTICAL METHODS 	  113
    8.1  ENVIRONMENTAL MEDIA 	  113
         8.1.1  Air 	  113
                8.1.1.1  Sample collection and preparation 	  113
                8.1.1.2  Methods 	  113
         8.1.2  Water 	  118
                8.1.2.1  Sample collection and preparation 	  118
                8.1.2.2  Methods 	  118
         8.1.3  Soil 	  118
                8.1.3.1  Sample collection and preparation 	  118
                8.1.3.2  Methods 	  118
         8.1.4  Food 	  118
                8.1.4.1  Sample collection and preparation 	  118
                8.1.4.2  Methods 	  118
                                                                       vil

-------
 Concencs

      8.2  BIOMEDICAL SAMPLES  	         119
           8.2.1  Fluids/Exudates  	'.'.'.'.'.'.'.'.'.'.'.'.'...'." 119
                  8.2.1.1  Sample preparation	  119
                  8.2.1.2  Methods 	 '.'.'.'.. 119
           8.2.2  Tissues  	 120
                  8.2.2.1  Sample preparation 	 120
                  8.2.2.2  Methods 	  120

  9.   REGULATORY AND ADVISORY STATUS 	                 121
      9.1  INTERNATIONAL 	','.',',	 i2i
      9. 2  NATIONAL	 121
           9.2.1  Regulations 	  121
                  9.2.1.1  Media-specific	 121
                  9.2.1.2  Hazard ranking 	' 121
                  9.2.1.3  Emission and effluent regulations 	 123
                  9.2.1.4  Consumer products regulations  	 124
           9.2.2  Advisory Guidance 	 124
                  9.2.2.1  Media-specific 	'.'.'.'.'.'.'.'.'.'. 124
           9.2.3  Data Analysis 	 125
                  9.2.3.1  Carcinogenic potency  	             125
      9.3   STATE 	"'' 125
           9.3.1  Regulations 	 125
                  9.3.1.1  Media-specific 	'.'.'.'.'.'.'.'.'.'.'.'. 125
           9.3.2  Advisory Guidance 	 126
 10.   REFERENCES 	 12y

 11.   GLOSSARY 	  169

 APPENDIX:  PEER REVIEW 	  173
viii

-------
                            LIST OF FIGURES
1.1  Health effects from breathing benzene 	    4
1.2  Health effects from ingesting benzene 	    5
2.1  Effects of benzene-- inhalation exposure 	    9
2.2  Effects of benzene--oral exposure 	   10
2.3  Levels of significant exposure for benzene-- inhalation 	   11
2.4  Levels of significant exposure for benzene--oral 	   12
2.5  Correlation of urinary phenol levels and atmospheric
     benzene concentrations in workers occupationally exposed
     in a rubber coating plant 	   19
2.6  Availability of information on health effects of benzene
     (human data) 	   23
2.7  Availability of information on health effects of benzene
     (animal data) 	   24
4.1  Biotransformation of benzene 	   41
4.2  Urinary metabolites of benzene 	   45
                                                                      ix

-------
                             LIST OF TABLES
3.1  Chemical Identity of benzene 	   32
3.2  Physical and chemical properties of benzene 	   33
4.1  Summary of results of some teratological studies on
     benzene in the mouse and rabbit 	   61
4.2  Teratology studies on inhaled benzene in rats 	   62
4.3  In vivo genotoxicity studies of benzene 	   68
4.4  In vitro genotoxicity studies of benzene 	   71
4.5  Case studies of workers occupationally exposed to benzene ....   74
4.6  Epidemiological studies of workers exposed to benzene 	   77
4.7  Carcinogenicity review studies of occupationally
     exposed workers 	   81
4.8  Summary of animal inhalation carcinogenicity experiments 	   86
4.9  Carcinogenic-related end points observed in animals
     exposed to benzene by inhalation 	   87
4.10 Summary of animal oral/gavage carcinogenicity experiments ....   88
4.11 Carcinogenicity-related end points observed in animals
     exposed to benzene by gavage 	   90
5.1  Disposal of petroleum industry wastes containing benzene 	   97
6.1  Annual emissions of benzene to air from various sources
     in the United States 	  100
6.2  Annual emissions of benzene to water in the United States ....  100
6.3  Benzene concentrations in wastewaters 	  101
7.1  Benzene levels in air samples 	  106
7.2  Benzene levels in water samples 	  109
7.3  Number of employees exposed to benzene (by exposure
     levels and by industry divisions)  	  Ill
8.1  Analytical methods for measuring benzene levels in air 	  114
8.2  Analytical methods for measuring benzene levels
     in water and soil 	  116
8.3  Analytical methods for measuring benzene levels in biological
     samples and food  	  117
9.1  Regulations and advisory guidance  for benzene  	  122
                                                                      XL

-------
                      1.  PUBLIC HEALTH STATEMENT
1.1  WHAT IS BENZENE?
     Benzene is a naturally occurring substance produced by volcanoes
and forest fires and present in many plants and animals, but benzene is
also a major industrial chemical made from coal and oil. As a pure
chemical, benzene is a clear, colorless liquid. In industry, benzene is
used to make other chemicals, as well as some types of plastics,
detergents, and pesticides. It is also a component of gasoline.

1.2  HOV MIGHT I BE EXPOSED TO BENZENE?

     The three main types of exposure to benzene are environmental,
consumer product, and occupational. Without question, the greatest
possibility for high-level exposures is in the workplace. However most
people are exposed to benzene in tobacco smoke and automobile exhaust.
     Benzene has been found in at least 337 of 1,177 National Priorities
List (NPL) hazardous waste sites. Other environmental sources of benzene
include gasoline (filling) stations, vehicle exhaust fumes, tobacco
smoke, underground storage tanks that leak, wastewater from industries
that use benzene, chemical spills, groundwater next to landfills
containing benzene, and possibly some food products that contain benzene
naturally. In addition, certain industries may.release benzene into the
surrounding air. These include ethylbenzene- and styrene-production
facilities, petroleum refineries, chemical manufacturing plants,  and
recovery plants for coke oven by-products. People living near such
industries may be exposed to benzene in the surrounding air.

     Consumer products containing benzene include glues, adhesives,
household cleaning products, paint strippers, some art supplies, tobacco
smoke, and gasoline.
     Occupational exposure to benzene can occur in the rubber industry;
oil refineries; chemical plants; the shoe manufacturing industry; and
gasoline storage, shipment, and retail stations.

1.3  HOV DOES BENZENE GET INTO MY BODY?

     Because benzene evaporates very quickly, the most common exposure
to benzene comes from breathing air containing benzene.

     Very small amounts of benzene are found in some foods, such as
canned beef, and in contaminated drinking water.
     Although benzene can enter through the skin, very few  individuals
come in contact with liquid benzene, except possibly through contact
with benzene-containing products such as gasoline.

-------
 2   Section 1

 1.4  UOV CAN BENZENE AFFECT MY HEALTH?

      Benzene is harmful,  especially to the  tissues  that  form blood
 cells.  How benzene affects your health would  depend on how much you are
 exposed to and how long you are exposed to  it.

 1.4.1  Brief Exposure at  High Levels

      Death may occur in humans and animals  after brief oral or
 inhalation exposures to high levels of benzene; however,  the  main
 effects of these types of exposures are drowsiness,  dizziness,  and
 headaches.  These symptoms disappear after exposure  stops.

 1.4.2  Long-Term Exposures at Various  Levels

      From overwhelming human evidence  and supporting animal studies,
 benzene is  known to be a  human carcinogen.  Leukemia  (cancer of  the
 tissues that form the white blood  cells) and  subsequent death from
 cancer  have occurred in some workers exposed  to benzene for periods  of
 less  than 5 and up to 30  years.  Long-term exposures  to benzene  may
 affect  normal blood production,  possibly resulting  in severe  anemia  and
 internal  bleeding.

      In addition,  human and animal studies  indicate  that benzene is
 harmful to  the immune system,  increasing the  chance  for infections and
 perhaps lowering the body's defense  against tumors.  Exposure  to benzene
 has also  been linked with genetic  changes in  humans and animals.
      Animal studies  indicate that  benzene has adverse effects on unborn
 animals.  These effects  include  low birth weight, delayed bone formation,
 and bone  marrow damage. Some of  these  effects occur at benzene  levels as
 low as  10 parts  of benzene  per million parts  of air  (ppm).  Although
 benzene has been reported to have  harmful effects on animal
 reproduction,  the  evidence  for human reproductive effects,  such as
 spontaneous abortion or miscarriage, is too limited to form a clear  link
 with  benzene.

 1.5   IS THERE  A  MEDICAL TEST TO  DETERMINE IF  I HAVE
      BEEN EXPOSED  TO BENZENE?

      Benzene can be  measured in  the  blood and the breath. The body
 changes benzene  to a chemical called phenol, which can be measured in
 the urine.  Amounts of benzene  (in  blood) and phenol (in urine) cannot be
used  as yet to predict what degree of harmful health effects may occur.

      The meaning of  benzene and  phenol measurements in blood and urine
 should be viewed carefully  for several  reasons: 1) phenol occurs
naturally in urine,  and urinary  amounts of phenol would have to be much
higher  than usual before  any measurement was meaningful;  2) present  test
methods are  limited  and raise doubts about the blood level values  found
 in some laboratories;  3)  because smoking can raise the background  level
of benzene  in  the blood,  smoking habits must be considered when
evaluating  exposure  to benzene;  4) benzene disappears rapidly from the
blood and measurements may  be accurate only for recent exposures;
5) average  amounts of benzene found  in  the body have not been determined
 for the general  population.

-------
                                             Public Health Statement   3

1.6  WHAT LEVELS OF EXPOSURE HAVE RESULTED IN HARMFUL HEALTH EFFECTS?

     The graphs on the following pages show the relationship between
exposure to benzene and known health effects. Effects in animals are
shown on the left side, effects in humans on the right.  The first
column, called "short-term exposure," refers to known health effects in
laboratory animals and humans from exposure to benzene for 14 days or
less. The second column, "long-term exposure," refers to benzene
exposures of more than 14 days.
     In the first set of graphs, labeled "Health effects from breathing
benzene" (Fig. 1.1), exposure is measured in parts of benzene per
million parts of air (ppm). The number of cases of cancer that could
occur after breathing 1 ppm benzene for a lifetime has been estimated to
be 260 persons in a population of 10 thousand, or 260 thousand persons
in a population of 10 million individuals. It should be noted that these
risk values are plausible upper-limit estimates. Actual risk levels are
unlikely to be higher and may be even lower.
     The levels marked on the graphs as "minimal risk for effects other
than cancer" show estimates of levels of exposure at which no adverse
effects are expected to occur. These levels are based on animal studies,
but some uncertainty still exists.
     In the second set of graphs, the same relationship is shown for the
known "Health effects from ingesting benzene" (Fig. 1.2). Exposures are
measured in milligrams of benzene per kilogram of body weight (mg/kg).
Although not enough information was available to estimate health effects
from absorbing benzene through the skin, benzene is known to enter
through the skin; this does not mean there is no possibility of a
hazard.

1.7  WHAT RECOMMENDATIONS HAS THE FEDERAL GOVERNMENT MADE TO
     PROTECT HUMAN HEALTH?
     The Environmental Protection Agency (EPA) set the maximum
permissible level in drinking water at 5 parts of benzene per billion
parts of water (ppb).  Because benzene can cause leukemia, EPA
established an ultimate goal of 0 ppb for benzene in drinking water and
in ambient water such as rivers and lakes. EPA realizes that this goal
may be-unattainable and has estimated how much benzene in ambient water
would be associated with one additional cancer case for every 100,000
persons (6.6 ppb benzene), one case for every 1 million persons (0.66
ppb benzene), and one case for every 10 million persons (0.066 ppb
benzene).
     Although more people are exposed to benzene outside the workplace
than in the workplace, the highest levels of benzene exposures occur  in
the workplace.
     The National Institute for Occupational Safety and Health  (NIOSH)
has recommended an occupational exposure limit  in air of 0.1 part of
benzene per million parts of air  (ppm). The Occupational Safety and
Health Administration's (OSHA) legally enforceable limit is an average
of 1.0 ppm over the standard 8-hour workday.

-------
Seccion  1
           SHORT-TERM EXPOSURE
       (LESS THAN OR EQUAL TO 14 DAYS)
                          LONG-TERM EXPOSURE
                         (GREATER THAN 14 DAYS)
EFFECTS
IN
ANIMALS
CONC IN
AIR
(Ppm)
EFFECTS
IN
HUMANS
EFFECTS
IN
ANIMALS
CONC IN
AIR
(ppm)
EFFECTS
IN
HUMANS
                  100.000
                              -DEATH
    DEATH -
                 •10.000
                  1.000
                                                            100000
                                                             10.000
                                                             1.000
EFFECTS ON
OFFSPRING

EFFECTS ON
BLOOD-
FORMING

EFFECTS ON4-'
IMMUNE I
SYSTEM
ru
. 1
EFFECT ON
REPRODUCTION. 	
LEUKEMIA
0 1(
1 f DROWSINESS
i t . .« » & * j». .^
f •< HEADACHE
> [ DIZZINESS
0 1

10
BLOOD-
FORMING
ORGANS
                   10
                   01
                  0 01-
 MINIMAL RISK
.FOR EFFECTS
 OTHER THAN
 CANCER
                                                              01
                                                             001
                                                                        MINIMAL RISK FOR
                                                                       • EFFECTS OTHER
                                                                        THAN CANCER
                 Fig. 1.1. Healtb effects from breathing benzene.

-------
                                                 Public Health  Statement
    SHORT-TERM EXPOSURE
(LESS THAN OR EQUAL TO 14 DAYS)
 LONG-TERM EXPOSURE
(GREATER THAN 14 DAYS)
EFFECTS EFFECTS
IN OOSE IN
ANIMALS (mg/kg/day) HUMANS
10.
11
1
1
000
DO
I0 	 DEATH
3
0
                                     EFFECTS
                                        IN
                                     ANIMALS
   DOSE
 (mg/kg/day)

   10.000
                                                   1000
                                                   100
                                     CANCER
                                     LOW WHITE
                                     BLOOD CELL
                                     COUNT
                                                  I 10
            01                                      01

                  Fig. 1.2.  Health effects from ingesting benzene.
     EFFECTS
        IN
     HUMANS

QUANTITATIVE DATA
WERE NOT AVAILABLE

-------
                       2.  HEALTH EFFECTS SUMMARY

2.1  INTRODUCTION

     This section summarizes and graphs data on the health effects
concerning exposure to benzene. The purpose of this section is to
present levels of significant exposure for benzene based on key
toxicological studies, epidemiological investigations, and environmental
exposure data. The information presented in this section is critically
evaluated and discussed in Sect. 4, Toxicological Data, and Sect. 7,
Potential for Human Exposure.

     This Health Effects Summary section comprises two major parts.
Levels of Significant Exposure (Sect. 2.2) presents brief narratives  and
graphics for key studies in a manner that provides public health
officials, physicians, and other interested individuals and groups with
(1) an overall perspective of the toxicology of benzene and (2) a
summarized depiction of significant exposure levels associated with
various adverse health effects. This section also includes information
on the levels of benzene that have been monitored in human fluids and
tissues and information about levels of benzene found in environmental
media and their association with human exposures.

     The significance of the exposure levels shown on the graphs may
differ depending on the user's perspective. For example, physicians
concerned with the interpretation of overt clinical findings in exposed
persons or with the identification of persons with the potential to
develop such disease may be interested in levels of exposure associated
with frank effects (Frank Effect Level, FEL). Public health officials
and project managers concerned with response actions at Superfund sites
may want information on levels of exposure associated with more subtle
effects in humans or animals (Lowest-Observed-Adverse-Effect Level,
LOAEL) or exposure levels below which no adverse effects (No-Observed-
Adverse-Effect Level, NOAEL) have been observed. Estimates of levels
posing minimal risk to humans (Minimal Risk Levels) are of interest to
health professionals and citizens alike.

     Adequacy of Database (Sect. 2.3) highlights the availability of  key
studies on exposure to benzene in the scientific literature and displays
these data in three-dimensional graphs consistent with the format in
Sect. 2.2. The purpose of this section is to suggest where there might
be insufficient information to establish levels of significant human
exposure. These areas will be considered by the Agency for Toxic
Substances and Disease Registry (ATSDR), EPA, and the National
Toxicology Program (NTP) of the U.S. Public Health Service in order to
develop a research agenda for benzene.

-------
 8   Section 2

 2.2   LEVELS OF SIGNIFICANT EXPOSURE

      To help public health professionals  address  Che  needs  of persons
 living or working near hazardous waste  sites,  the  toxicology  data
 summarized in this section are organized  first by  route  of  exposure--
 inhalation,  ingestion,  and dermal--and  then by toxicological  end points
 that are categorized into six general areas--lethality,  systemic/target
 organ toxicity,  developmental toxicity, reproductive  toxicity,  genetic
 toxicity,  and carcinogenicity.  The  data are discussed in terras  of  three
 exposure periods--acute,  intermediate,  and chronic.

      Two kinds of graphs  are used to depict the data. The first type is
 a "thermometer11  graph.  It provides  a graphical summary of the human and
 animal toxicological end  points (and levels of exposure)  for  each
 exposure route for which  data are available. The ordering of  effects
 does not reflect the exposure duration  or species  of  animal tested. The
 second kind of graph shows Levels of Significant Exposure (LSE) for each
 route and exposure duration.  The points on the graph  showing  NOAELs and
 LOAELs reflect the actual doses (levels of exposure)  used in  the key
 studies.  No  adjustments for exposure duration  or intermittent exposure
 protocol were  made.

      Adjustments reflecting the uncertainty of extrapolating  animal data
 to humans,  intraspecies variations, and differences between experimental
 vs actual  human  exposure  conditions were  considered when estimates of
 levels posing  minimal  risk to human health were made  for noncancer end
 points.  These  minimal  risk levels were  derived for the most sensitive
 noncancer  end  point  for each exposure duration by  applying uncertainty
 factors. These levels  are shown on  the  graphs  as a broken line  starting
 from the actual  dose  (level of  exposure)  and ending with a concave-
 curved line  at its  terminus.  Although methods  have been  established to
 derive these minimal risk levels (Barnes  et al. 1987), shortcomings
 exist  in the techniques that reduce the confidence in the projected
 estimates. Also  shown  on  the  graphs under the  cancer end point  are low-
 level  risks  (1(T4  to 10'7)  reported by EPA.  In addition, the  actual dose
 (level of  exposure)  associated  with the tumor  incidence  is plotted.

 2.2.1   Key Studies and  Graphical Presentations

      For benzene,  the  "thermometer" graphs for inhalation exposure are
 shown  in Fig. 2.1  and  those for oral exposure  are shown  in Fig. 2.2. The
 corresponding LSE  graphs  are  shown  in Figs.  2.3 and 2.4. Data were not
 found  for  deriving LOAELs and NOAELs based on  the toxic  effects of
benzene via dermal absorption.

 2.2.1.1  Lethality

      Inhalation.  Acute lethality data  in humans are  limited  to case
 studies, mostly  of victims  of inhalation  toxicity, in which exposure
 levels  are generally not  known  but have been estimated to be  19,000 to
 20,000 ppm for 5 to  10  min (Sandmeyer 1981). These data  suggest a
possible range for the  LOAEL for acute lethality in humans.  In  the rat,
 the  LOAEL  for an LCso value can be estimated to be 13,700 ppm for a 4-h
exposure (Drew and Fouts  1974).  A slightly higher mortality rate (4/6)
has been reported  for a 4-h exposure to 16,000 ppm (Smyth et  al. 1962).
These  data are indicative of  relatively low acute toxicity.

-------
                                                                   Health  Effects Summary     9
  4NIVWLS
    Cpni
10000
 1000
                                                                        'SO")

                                                                    .00000
             RAT  .Cw «•> COS'NUOLS
                                                                      1000 -
     -I
          •  RABBIT DEVELOPMENTAL TOXICITV \ 3 OAVS CONTINUOUS
          •  MOUSE LYMPWOMA  16 WEEKS INTERMITTENT
          •  MOUSE REPRODUCTIVE TOXIC.TV • 3 WEE
-------
10     Section 2
     ANIMALS
    (mg/kg/day)

  10000 i-
  1000
    100
    10
                                                                       HUMANS
                                                                      (mg/kg/day)

                                                                   10000 |-
                                                                           1.000
          • RAT LD,
                                                                            100
   • RAT. CARCINOGENICITY. 103 WEEKS. INTERMITTENT


   • MOUSE. CARCINOGENICITY. 103 WEEKS. INTERMITTENT




-  • RAT HEMATOTOXICITY. 6 MONTHS. INTERMTTTENT
                                                                             10
         O RAT. HEMATOTOXICITY. 6 MONTHS. INTERMITTENT

              • LOAEL IN ANIMALS         A LOAEL IN HUMANS
              O NOAEL IN ANIMALS

                         Fig. 2.2. Effects of benzene—oral exposure.
                                                                              1L-
                                                                                     OEATH

-------
                                                          Health Effaces Summary    1L
                        ACUTE
                      (SI4 DAYS)
                            INTERMEDIATE
                            (15-364 DAYS)
                       DEVELOP-   TARGET
            LETHALITY  MENTAL   ORGAN
                    TARGET           REPRO-
                    ORGAN  CANCER  DUCTION
   (ppm)


  100000



  10.000



   1.000



    100



     10
     01
    001
   0001
  00001
 0 00001
0 000001
r
          (BONE
• m      MARROW)
   . (CNS)
               r
m (BONE MARROW)   9 m
       • m
            • m (BONE MARROW)
                             CHRONIC
                           (2365 DAYS)
                             CANCER
                                                                       10~* -i
                                                                         ,-S-
                                                                       10
                                                                       10~6 ~
                                                                        10
                                                                         ,-7-1
                                                       ESTIMATED
                                                       UPPER-BOUND
                                                       HUMAN CANCER
                                                       RISK LEVELS
             • LOAEL FOR ANIMALS
             O NOAEL FOR ANIMALS
             A LOAEL FOR HUMANS
             A NOAEL FOR HUMANS
                  r   RAT
                  m  MOUSE
                  n  RABBIT
              MINIMAL RISK
              FOR EFFECTS
              OTHER THAN
              CANCER
                    Fig. 2.3. Levels of significant exposure for benzene—inhalation.

-------
12   Section 2
   (mg/kg/day)


    10000 i-



     1.000



      100



       10
      0 1
     001
    0001
   00001
  000001 -
 0 000001 •-
                   ACUTE
                 (S14DAYS)
                  LETHALITY
          • LOAEL FOR ANIMALS
          O NOAEL FOR ANIMALS
          A LOAEL FOR HUMANS
  INTERMEDIATE
  (15-364 DAYS)

 TARGET ORGAN
r (BONE MARROW)
        CHRONIC
       (^365 DAYS)

        CANCER
    RAT
                                                      10-41
                                                       -5-
                                                      10
                                                      10-6-
                                                       -7-1
                           ESTIMATED
                           UPPER-BOUND
                           HUMAN CANCER
                           RISK LEVELS
                                                      10
MINIMAL RISK
FOR EFFECTS
OTHER THAN
CANCER
              Fig. 2.4.  Lereb of significant exposure for benzene—oral.

-------
                                             Health Effects Summary   13

     Oral.  Oral lethal doses for humans have been estimated at 10 mL
(8.8 g) (Thienes and Haley 1972, as reported in Sandmeyer 1981), 9 to
12 g (Von Oettingen 1940, as reported in Sandmeyer 1981), and 30 g
(Moeschlin 1965, as reported in Sandmeyer 1981). After conversion to
milligrams per kilogram (for a 70-kg adult), a range of 128 to 428 mg/kg
can be estimated for oral lethal exposures in humans. On Figs. 2.2 and
2.4, 128 mg/kg is plotted as an estimated LOAEL for lethality in humans.
For the rat, the lowest LDSO found in the literature was 930 mg/kg; this
value is plotted on Figs. 2.2 and 2.4 as a LOAEL for LD50 in animals.
     Dermal.  No data were found for lethality of benzene in humans or
animals by dermal exposure.

2.2.1.2  Systemic/target organ tozicity

     The most sensitive target systems for benzene toxicity are the
hematopoietic and the immune systems. The nervous system is also
important in the context of acute toxicity.

     Numerous studies in animals have shown that benzene-induced bone
marrow depression is the result of damage to the pluripotential stem
cells and/or the early proliferating committed cells in either erythroid
(red cell) or myeloid (white cell) lines (Toft et al. 1982, Green et.
al. 1981). These alterations can occur during short-term exposures to
-10 ppm benzene. Similar adverse effects have also been observed in some
workers exposed to low levels of benzene for short time periods. Because
bone marrow depression is generally associated with long-term exposure,
a "sensitization phenomenon," occurring in a particularly susceptible
population, has been proposed as a possible reason for the effects
observed following low-level, short-term exposures (Aksoy et al. 1976,
Ikeda 1964). These short-term effects may also be related to the
accumulation of benzene (metabolized in the liver and in the bone
marrow) and its putative toxic metabolites in the bone marrow (Rickert
et al.  1979; Irons et al. 1980a,b)

     Inhalation, acute.  LOAELs and NOAELs have been derived for what
appear to be the most sensitive parameters of hematotoxicity,
immunotoxicity, and neurotoxicity.

     Toft et al. (1982) exposed NMRI mice intermittently for 8 h/day,
5 days/week, for 2 weeks to benzene concentrations ranging from 1 to
200 ppm. The LOAELs in this study were 50 ppm (P £ 0.05) for decreased
bone marrow cellularity (decreased number of nucleated cells/tibia) and
21 ppm (P £ 0.05) for decreases in the number of granulopoietic stem
cells in the marrow of the tibia. With the same doses administered
continuously, the LOAEL was 21 ppm for both parameters. NOAELs were not
clear in this study.

     The effects in the bone marrow are reflected in the peripheral
blood cell counts. Li et al. (1986) examined the effect of benzene and
other solvent vapors on peripheral blood leukocytes and leukocyte
alkaline phosphatase levels in female Wistar rats. The animals were
exposed to 0, 20, 50, 100, 300, 1,000, or 3,000 ppm benzene, 8 h/day  for
7 days, and leukocyte enzyme activities and leukocyte counts were
determined. The leukocyte counts were depressed significantly (based  on
nonoverlapping standard deviations) at 50 ppm (LOAEL) and not

-------
14   Section 2

significantly at 20 ppm (NOAEL). At 300 ppm, the enzyme levels were
significantly increased (P < 0.01) (LOAEL),  and at 100 ppm the levels
were increased but not significantly (P > 0.10) (NOAEL). There were no
effects at SO ppm.
     Rozen et al. (1984) observed dose-dependent depressions of
peripheral red blood cell (RBC) and lymphocyte counts in CS7B16 male
mice exposed to benzene concentrations of 0, 10.2, 31.0, 100, or 301 ppm
6 h/day for 6 days. The LOAEL for RBC depression was 100 ppm; the NOAEL
was 30 ppm. Lymphocyte counts were depressed at all levels (P < 0.05);
thus the LOAEL for lymphocyte depression was 10.2 ppm. These results are
consistent with the observation that, of the erythroid, myeloid,  and
lymphoid blood cell lineages, the lymphoid line appears to be the most
sensitive to benzene toxicity (Goldstein 1977, Irons et al. 1979).
Lymphocytes play an important role in the immune response.
     To measure cellular immunity in mice, a function dependent on
lymphocyte populations, Rosenthal and Snyder (1985) tested host
resistance to the bacterium Listeria aonocytogenes during 12 days
(6 h/day) of exposure to benzene. The benzene concentrations tested were
10, 30, 100, and 300 ppm. The numbers of bacteria in the host spleens
increased significantly (P s 0.05) at 30 ppm (LOAEL) or greater;  there
was an effect at 10 ppm, but it was not statistically significant
(NOAEL).

     Based on data presented by Gerarde (1959, 1960) and Von Oettingen
(1940, as reported in Sandmeyer 1981), Sandmeyer (1981) correlated the
signs and symptoms of acute benzene toxicity in humans via inhalation
with concentration and duration of exposure: 1.5 ppm was the olfactory
threshold; 25 ppm for 480 min had no obvious effect, even though benzene
was detectable in the blood; 50 to 150 ppm for 300 min produced
headache, dizziness, and lassitude; 500 ppm for 60 min produced
headache; 1,500 ppm for 60 min caused signs  of illness; 3,000 ppm was
tolerated for 30 min to 1 h; 7,500 ppm induced signs of toxicity in
30 min to 1 h; and 19,000 to 20,000 ppm for  5 to 10 min may be fatal.

     At inhalation exposures of less than 100 ppm, men and women absorb
-50% of the dose in 4 h (Nomiyama 1974a,b, both as reported in IARC
1982). However,  this fraction absorbed appears to increase with
decreasing dose.

     Inhalation, Intermediate.  CD-I mice were exposed to 1, 10,  30, and
300 ppm benzene vapor 6 h/day, 5 days/week,  for 13 weeks (Ward et al.
1985). Interim examination on day 28 of the  study revealed statistically
significant (P < 0.05) hematological changes in the males and females of
the 300-ppm group. These changes included decreased erythrocyte and
leukocyte counts. Treatment-related changes  were not observed at lower
concentrations.  The LOAEL for these effects  is, therefore, 300 ppm, the
NOAEL 30 ppm.

     Inhalation, chronic.  Data were not sufficient to derive LOAELs and
NOAELs.

     Oral, acute.  No data were found from which to derive LOAELs and
NOAELs.

-------
                                             Health Effects Summary   L5

     Oral,  intermediate.   Wolf et  al.  (1956) conducted a 6-month study
 in Wistar rats  using benzene  doses of  0,  1,  10,  SO, and 100 mg/kg/day
 and evaluated hematological parameters.  Leukopenia was a dose-related
 effect which was  slight at 10 mg/kg/day  and  not  observed at 1 mg/kg/day.
 The data were not treated statistically.  A LOAEL of 10 mg/kg/day and a
 NOAEL of 1  mg/kg/day were identified for this study.

     Oral,  chronic.  Data were not found from which to derive LOAELs and
 NOAELs.

     Dermal.  No  data were found for hematological effects via dermal
 exposure for acute, intermediate,  or chronic exposure.

 2.2.1.3  Developmental toxicity

     A number of  investigations have evaluated the
 developmental/maternal toxicity of benzene in animals. They have
 demonstrated that benzene is  not teratogenic but does cause maternal
 toxicity and embryo/fetotoxicity,  sometimes  at levels as low as 10 ppm.
 As will be briefly discussed  in the following paragraphs, the
 determination of  minimal  risk from current information has many
 uncertainties.

     Inhalation.  With respect to  decreased  fetal weight and skeletal
 variations,  only  the data for inhalation exposures are adequate to
 determine LOAELS  and NOAELS and minimal  risk levels.

     The study of Ungvary and Tatrai (1985)  demonstrates dose-dependent
 fetotoxic effects in mice exposed  on days 6  to 15 of gestation and
 rabbits exposed on days 7 to  20 of gestation to benzene concentrations
 of 156 and 313 ppm. In both cases,  exposure was continuous (24 h/day).
 The LOAEL for fetal weight and skeletal  variations in rabbits was
 313 ppm, and the  NOAEL was 156  ppm. For  mice, the LOAEL is estimated Co
 be 156 ppm,  but lower concentrations were not tested. The study appeared
 to be well conducted, but the  use  of only two concentrations limits the
 conclusions that  can be reached.

     Kuna and Kapp (1981)  tested three concentrations of benzene on
 pregnant rats 7 h/day, during days  6 to  15 of gestation. The LOAEL for
 fetal weight and  skeletal variations was  50 ppm, and the NOAEL was
 10 ppm.  The investigators stated that the chemical showed potential, but
 not statistically significant,  teratogenicity at 500 ppm.

     Oral.   No data were  found from which to derive LOAEL and NOAEL
values for oral developmental  toxicity;  however, Seidenberg et al.
 (1986) observed reduced fetal  body weights in the offspring of mice that
were given 1,300 mg/kg of benzene  per day by gavage on gestation days 8
 to 12. In addition, Nawrot and Staples (1979) reported an increase in
 resorptlons in pregnant CD-I  mice  given  oral doses of 0.5 or
 1.0 mLAg/day (both of which  were  maternally toxic), but not at
 0.3 mLAg/day.

     Dermal.  No  data were found for dermal  developmental toxicity.

-------
 16    Section  2

 2.2.1.4  Reproductive  toxicity

      Inhalation.  Ward et  al. (1985) exposed mice to benzene
 concentrations  of 1, 10, 30,  or  300 ppm  for 13 weeks. A LOAEL of 300 ppra
 was  noted  for histopathological  changes  in the ovaries (bilateral cysts)
 and  testes (atrophy/degeneration, decrease in spermatozoa, moderate
 increase in abnormal sperm forms). The NOAEL for these effects was
 30 ppm. The difference between the values considered to be the NOAEL and
 the  LOAEL  was tenfold;  more closely spaced doses could reveal these
 values to  be  closer.

      Oral.  Data  from  which to derive LOAELs and NOAELs for reproductive
 toxicity by oral  administration  of benzene were not found.

      Dermal.  Data were not found for reproductive toxicity of benzene.

 2.2.1.5  Carcinogenicicy

      Inhalation.  Benzene  is  considered  to be a human carcinogen by EPA,
 OSHA, the  World Health Organization (WHO), and the International Agency
 for  Research  on Cancer (IARC).

      The EPA  (1986) has reviewed the human and animal carcinogenicity
 data on benzene,  and this  report should be consulted for data of
 interest (e.g., q.* values for each of the oral and inhalation animal
 studies reviewed). The  following paragraph is a summary of the EPA
 calculations  of unit risk values for leukemia based on human
 epidemiological studies. It should be noted that these values are
 estimates  of  human risk, since the true human risk at low doses cannot
 be accurately identified.

      The potency  of benzene has  been estimated based on three separate
 epidemiological studies (Rinsky  et al. 1981;  Ott et al. 1978; Wong et
 al.  1983,  as  reported  in EPA  1986). Giving equal weight to cumulative
 dose  and weighted cumulative  dose, as well as relative and absolute
 model forms,  EPA  estimated a  risk value of 2.6 x 10*2 for leukemia due
 to a  lifetime exposure  of  1 ppm  benzene  in the air (EPA 1986). Based on
 this  value, the exposure levels  associated with individual lifetime
 upper-bound risks of ID""*, 10*^, 10*6, an
-------
                                              Health  Effects Summary   L7

      In  another case  history,  a chemical  plant  worker  died of  "acute
 myelogenous  leukemia" at age  51,  15  years after being  exposed  for an
 18-month period to  benzene  concentrations of  less  than 2 ppm,  time-
 weighted average (TWA)  (Ott et al. 1978).  These two  data points are
 indicated  in Fig. 2.3.

      In  animal  studies  it is  noteworthy that, in one bioassay,
 relatively short-term exposure to benzene (300  ppm for only 16 weeks)
 was  leukemogenic in mice 1.5  years later  (Cronkite 1986). This duration
 of exposure  is  less than that  for the usual chronic  carcinogenicity
 bioassay,  and,  to indicate  this  fact, the data  point has been placed in
 the  intermediate-exposure category in Fig. 2.3.

     Oral.   There are no oral  exposure data for benzene carcinogenicity
 in humans; however, the  oral dose levels  associated with specific
 carcinogenic  risks  can be extrapolated by converting the risk value of
 2.6  x 10'2 for  an inhalation exposure of  1 ppm  to  2.9  x 10'2 for an oral
 exposure of  1 mg/kg/day  and assuming identical  levels  of absorption of
 benzene  into  the body. Using the method described  in EPA (1986), the
 dose levels  associated with individual upper-bound estimates of risk of
 10'4, 10'5,  10'6, and 10'7  have been calculated to be  3.6 x 10'3,
 3.6 x 10 *4,  3.6  x 10-5.  and 3.6 x 10"6 mgAg/day,   respectively. (These
values are partially  based  upon Rinsky 1981 data.   The Rinsky 1987 data
have not been evaluated  by  EPA.)

     Rats administered benzene by gavage  at doses  of 25, 50,  and
 100 mg/kg/day for 103 weeks developed tumors of the zymbal gland and the
mouth at 50 mg/kg/day (NTP  1986). Mice treated  similarly developed
 lymphoma at 25 mg/kg/day. Other types of  tumors observed in this study
are described in a  later  section.

     Dermal.   Data were  not found for the carcinogenicity of benzene
administered by  the dermal  route, except  for many  skin painting studies
 in which benzene was used as a vehicle and was  negative for
carcinogenicity.

2.2.2  Biological Monitoring as a Measure of Exposure and Effects

     Biological monitoring  for benzene is complicated by the
difficulties in establishing precise correlations between the monitoring
end points, the equivalent  exposure levels, and the ultimate toxic
effects.  Most monitoring  end points provide only a rough estimate of
exposure because of the potentially wide variability in background
levels.  Current methods  lack sensitivity at levels corresponding to
exposures below about 10  ppm (inhalation exposure). New methods would
have to be developed before biological monitoring could be used to
identify minimal effect  levels.

     Biological monitoring  can be based on indirect indicators of
exposure as determined by measurements of benzene  or benzene metabolites
in biological media, or  it  can be based on measurements of biological
effects induced by benzene.

2.2.2.1  Monitoring of exposure

     In monitoring for benzene exposure, both selective and nonselective
end points have been used.  Nonselective end points provide only a rough

-------
18   Section 2

estimate of exposure. One such nonselective test is the urinary sulfate
ratio test, which is based on the premise that with increasing exposure
there will be an increase in benzene metabolites conjugated with sulfate
moieties. Estimates of benzene exposure can be made by comparing the
ratio of inorganic to organic sulfates in the urine.  Inorganic sulfate
levels amounting to 80 to 95% of total urinary sulfates are considered
normal background, 70 to 80% indicate some exposure to benzene,  60 to
70% indicate a dangerous level of exposure, and 0 to 60% indicate an
extremely hazardous exposure (Hammond and Herman 1960). Urinary sulfate
levels are, however, quite variable, and they have not been used to
identify exposure levels associated with minimal toxic effects.

     As a urinary metabolite of benzene, phenol provides a more specific
indicator of exposure than do urinary sulfates. Phenol measurements have
routinely been used for monitoring occupational exposures (NIOSH 1974),
and there is evidence that urinary phenol levels can be correlated with
exposure levels (Pagnotto et al. 1961, see Fig. 2.5). Inoue et al.
(1986) reported a good correlation between occupational benzene
exposures and urinary phenol levels when expressed in terms of
creatinine excretion. The calculated mean correlation coefficient was
0.891. After reviewing the available data, NIOSH (1974) concluded that a
urinary phenol level of 75 mg/L provided a good correlation with an 8-h
TWA exposure to 10 ppm. However, according to studies reported on by
Lauwerijs (1979, as reported in van Sittert and de Jong 1985), this
exposure level corresponds to a urinary phenol level of 45 to 50 mg/L.
For exposures to 25 ppm benzene, urinary phenol levels of 100 to as high
as 200 mg/L have been reported (Sandmeyer 1981, NIOSH 1974).
     Correlating urinary phenol with benzene exposure is complicated by
potentially high background levels in nonexposed persons. Lauwerijs
(1979, as reported in van Sittert and de Jong 1985) reported that
nonexposed persons had urinary phenol levels of about 20 mg/L, and in
studies cited by NIOSH (1974), levels ranging from 5 to 42 mg/L have
been measured in nonexposed persons. Because of such high background
levels, benzene exposures of 5 ppm or less would be difficult to monitor
by measuring urinary phenol levels, and consequently, correlations
between phenol levels and no-observed-effect levels (NOELS) or NOAELS
would not be possible for effects occurring in this dose range.

     Preliminary results of studies in which determinations of benzene
in blood were compared with those of phenol in the urine shoved that
blood levels of benzene are more reliable for assessing both exposure
and uptake of benzene (Braier er al. 1981). However, because of the
short half-life of benzene in the blood and the observations in animal
studies that the rate of disappearance of benzene from the blood varies
according to the number of times the animal has been exposed (C. A.
Snyder et al. 1981c), blood levels of benzene may not be useful except
for recent exposures (Goldstein 1986).

     Urinary phenol excretions can be increased by exogenous non-
benzene sources such as dietary protein (Folin and Denis 1915);
medicines that contain phenylsalicylate (Pepto-Bismol* and Chloraseptic*
lozenges) (Kociba et al. 1976); aspirin (Fishbeck et al. 1975); and
calamine lotion and phenol-camphor-liquid petrolatum preparations
(Ruedemann and Deichmann 1953).

-------
                                                    Health Effects Summary
                                                                            19
     700  -
     600  -
£   500  -
O
z
111
X
Q.
z
cc
400   -
     300  -
200  -
     100
SPREADERS   O



SATURATORS   •



CHURN MEN   9
                 10     20     30    40     50    60     70     80

                             ATMOSPHERIC BENZENE (ppm)
                                                                       90
                                    100
       •According to NIOSH (1974), values given represent both phenol and parcresol. Phenol alone

    would result in values lower than indicated.

    Source: Pagnotto et al. 1961.
       Fig. 2.5.  Correlation of urinary phenol levels and atmospheric benzene concentrations in workers

    occnpatioomlly exposed in a rubber coating plant.

-------
 20   Section 2

      Because benzene is partially excreted in expired air, breath levels
 have been used as a measure of exposure. In a review of several
 occupational exposure studies, NIOSH (1974) reported that the benzene
 concentration in the breath was 2 ppm immediately following a 4.5-h
 exposure to 25 ppm and 60 to 70 ppm following 1- to 4-h exposures to
 100 ppm. In studies on humans, Hunter (1968)  found that benzene could be
 detected in expired air 24 h after a 100-ppm exposure and suggested that
 it might be possible to back-extrapolate to the concentration in the
 inspired air.  However,  the amount of benzene  lost in expired air will
 vary not only with the dose, but also with the extent of metabolism in
 the body.  Consequently, levels in the breath  may not be proportional to
 the dose.

      Little data are available correlating breath levels with exposures
 to very low concentrations of benzene in the  air.  Comparative studies of
 residents  in urban and rural areas have  revealed higher levels of
 benzene in the expired air of urban dwellers  (Wester et al.  1986).  For
 nonsmokers,  benzene breath levels were 2.5 ±  0.8 ppb in the  urban area
 and 1.8 ±  0.2  ppb in the rural area.  However,  in both cases  breath
 levels  were higher than ambient air levels (1.4 ± 0.1 and 1.0 ± 0.1  ppb,
 respectively),  suggesting that other sources  of exposure were occurring.

      One early animal study found that a linear relationship existed
 between the equilibrium concentration of benzene in the blood and the
 concentration  in the air (Schrenk et al.  1941).  A steady-state
 concentration  was reached in a few hours.  Because  blood levels generally
 decrease rapidly following exposure,  monitoring would have to be
 conducted  during exposure.  No information was  found correlating effects
 with  blood levels.  However,  blood levels  would be  expected to provide a
 more  accurate  assessment of internal  dose and,  thus,  a more  accurate
 prediction of  target organ effects than  other  monitoring end points.

 2.2.2.2  Monitoring of  effects

      In  addition to using benzene  and benzene  metabolite levels for
 monitoring purposes,  various biological  indices  might also be used  to
 measure  low-level exposures.  Monitoring  of benzene  workers has inr uded
 monthly  blood  counts, with workers being  removed from areas  of po?. -ntial
 exposure when white blood cell  (WBC)  counts fell below 5,000 or RBC
 counts fellbelow 4.000,000 (ITII  1975).  Van Sittert  and de  Jong (1985)
 have  suggested  that,  for some compounds  such as  benzene,  the biological
 end points of chromosomal  aberrations  in  peripheral  lymphocytes and
 sister chromatid exchanges  (SCEs)  could be  used  as  monitoring end
 points.

 2.2.3  Environmental  Levels  as  Indicators  of Exposure and  Effects

 2.2.3.1  Levels  found in the environment

     Benzene is  ubiquitous  in the  environment, and  a  large segment of
 the U.S. population is undoubtedly exposed, but  levels  are ordinarily
 quite low compared  with  occupational  exposure  levels.  There  are  no data
 documenting health  effects  as a result of  environmental  exposure  to
benzene, including  consumption of  certain  foods  known to contain
benzene.

-------
                                             Health Effects Summary   21

2.2.3.2  Human exposure potential

     For the majority of the U.S. population, the most likely route of
exposure to benzene is inhalation. Furthermore,  there is a much greater
risk of developing health effects from occupational exposure to benzene.
The manner of benzene disposal at waste sites will determine the most
significant route of human exposure. If it is buried, then the most
likely route will be through consumption of contaminated water supplies.
Whether the benzene reaches the groundwater will depend on many factors
(e.g., groundwater depth, soil type, bacterial population, and amount of
rainfall).  If benzene is disposed of in such a manner that it comes into
contact with air, inhalation then becomes an important exposure route.
Dermal absorption through contact with surface soils is not expected to
be significant, because of benzene's relatively high volatility.

     Regardless of the exposure route, the most important factor
affecting toxicity is systemic absorption. Only a portion of the dose of
a chemical to which one is exposed is systemically absorbed, and unless
local irritation at the route of entry is significant, this is the
fraction of concern. Toxicity studies with benzene have shown that
pulmonary irritation is not a problem except at very high exposure
levels, and the fraction systemically absorbed as a result of inhalation
may be less than 50% of the exposure concentration. The breathing rate
can be an important factor, since more benzene will be inhaled during
heavy labor than during rest.
     Because some, if not most, health effects of benzene are thought to
be due to the formation of its metabolites, its toxicokinetic pattern is
of interest. The fraction of the amount systemically absorbed that
undergoes biotransformation to reactive metabolites determines the
magnitude of the toxic response.

2.3  ADEQUACY OF DATABASE

2.3.1  Introduction
     Section 110 (3) of SARA directs  the Administrator of ATSDR to
prepare a toxicological profile  for each of  the 100 most  significant
hazardous substances found at facilities on  the CERCLA National
Priorities List. Each profile must  include the following  content:
    "(A)  An examination, summary,  and interpretation of  available
          toxicological  information and epidemiologic evaluations on a
          hazardous substance in order to  ascertain  the levels  of
          significant human exposure  for  the  substance and  the
          associated acute, subacute, and  chronic  health  effects.

      (B)  A determination of whether  adequate information on  the health
          effects  of each substance is available or  in  the  process  of
          development  to determine  levels  of exposure which present a
          significant  risk  to human health of acute,  subacute,  and
          chronic  health effects.
      (C)  Where  appropriate,  an identification of  toxicological testing
          needed to identify  the types or levels of  exposure  that  may
          present  significant  risk  of adverse health effects  in humans."

-------
 22    Section  2

      This  section  Identifies  gaps  In current knowledge relevant  to
 developing levels  of significant exposure for benzene. Such gaps are
 Identified for certain health effects end points (lethality,
 systemic/target organ toxicity, developmental toxiclty, reproductive
 toxiclty,  and carcinogenicity)  reviewed In Sect. 2.2 of this profile In
 developing levels  of significant exposure for benzene, and for other
 areas such as human biological  monitoring and mechanisms of toxicity.
 The present section briefly summarizes the availability of existing
 human and  animal data, identifies  data gaps, and summarizes research in
 progress that may  fill such gaps.

      Specific research programs for obtaining data needed to develop
 levels of  significant exposure  for benzene will be developed by ATSDR,
 NTP,  and EPA  in the future.

 2.3.2 Health Effect End Points

 2.3.2.1  Introduction and graphic  summary

      The availability of data for health effects in humans and animals
 is depicted on bar graphs in  Figs.  2.6 and 2.7,  respectively.

      The bars of full height  indicate that there are data to meet at
 least one  of  the following criteria:

 1.   For noncancer health end points, one or more studies are available
      that  meet current scientific  standards and are sufficient to define
      a range  of toxicity from no effect levels (NOAELs) to levels that
      cause  effects (LOAELs or FELs).

 2.   For human carcinogenicity, a substance is classified as either a
      "known human carcinogen" or a "probable human carcinogen" by both
      EPA and  IARC  (qualitative), and the data are sufficient to derive a
      cancer potency factor (quantitative).

 3.   For animal carcinogenicity, a substance causes a statistically
      significant number of tumors  in at least one species and the data
      are sufficient to derive a cancer potency factor.

 4.   There  are studies which show  that the chemical does not cause this
      health effect via this exposure route.

      Bars of half height indicate  that "some" information for the end
point exists but does not meet any of these criteria.

      The absence of a column  indicates that no information exists for
that  end point and route.

      Figures  2.6 and 2.7 summarize the adequacy of the existing database
for the end points of benzene toxicity that include lethality,
systemic/target organ toxicity, developmental toxicity, reproductive
toxicity, and carcinogenicity. Figure 2.6 represents human data;
Fig.  2.7 represents animal data. Each figure depicts the adequacy of the
available data for these effects by the inhalation, oral, and dermal
routes of exposure. The information on systemic effects is divided
according  to exposure duration. Acute exposure refers to exposures
lasting 14 days or less;  intermediate exposure refers to exposures

-------
                                     HUMAN DATA
                                                                                                           SUFFICIENT
                                                                                                          INFORMATION*
                                                                                                        V    SOME
                                                                                                        ^INFORMATION
                                                                                                      J
                                                                                                                NO
                                                                                                           INFORMATION
                                                                                       DERMAL
                                                                                                                            n
                                                                                                                            sr
                                                                                                                            0>
                                                                                                                            0
LETHALITY        ACUTE     INTERMEDIATE    CHRONIC   DEVELOPMENTAL  REPRODUCTIVE  CARCINOOENICITY
          Z	.	__/    TOXICITV        TOXICITY
                    SYSTEMIC TOXICITV

                      'Sufficient information exists to meet at least one of the criteria for cancer or noncancer end points
                           g. 2.6. Availability of information on health effects of benzene (human data).

-------
                                             ANIMAL  DATA
                                                                                                               SUFFICIENT
                                                                                                              INFORMATION'
                                                                                                                                 05
                                                                                                                                 r>
Q

NO
                                                                                                                 SOME
                                                                                                              INFORMATION
                                                                                                                  NO
                                                                                                              INFORMATION
                                                                                              INHALATION
                                                                                         DERMAL
LETHALITY
               ACUTE
                         INTERMEDIATE
                                       CHRONIC   DEVELOPMENTAL REPRODUCTIVE   CARCINOGENIC!! Y

                                            .—/     TOXICITY        TOXICITY
                    SYSTEMIC TOXICITY
                   'Sufficient information exists to meet at least one of the criteria tor cancer or noncancer end points.

                       Fig. 2.7. Availability of information on health effects of benzene (animal data).

-------
                                             Health Effects Summary   25

lasting 15 Co 364 days; and chronic exposure refers to exposures of
1 year or longer.

2.3.2.2  Descriptions of highlights of graphs

     Figure 2.6 shows that there are no human studies that qualify as
adequate for determining NOAELs, LOAELs, and FELs for benzene.
Inhalation carcinogenicity data are considered to be adequate by the EPA
for the estimation of a unit risk. Some data exist in the area of
lethality by the inhalation and oral routes, and some data exist for
acute and chronic systemic effects by the route of inhalation; however,
these are not sufficient for establishing NOAELs, LOAELs, and FELs. No
data were available for any systemic effects resulting from dermal
exposure; no data were available for systemic, developmental,
reproductive, or carcinogenic effects resulting from oral exposure; and
no data were available for systemic (intermediate exposure),
developmental, or reproductive effects resulting from inhalation
exposure to benzene.

     Figure 2.7 shows that adequate animal studies are available from
which to determine NOAELs, LOAELs, and FELs for systemic effects (acute
exposure) and developmental toxicity via inhalation and for systemic
effects (intermediate exposure) via oral exposure. Oral and inhalation
carcinogenicity data for benzene-exposed animals are adequate for
determining unit risk estimates for humans. Some data exist for
lethality by both the inhalation and oral routes, for reproductive
effects by inhalation, and for developmental toxicity via oral exposure,
but none are sufficient to determine NOAELs, LOAELs, and FELs for
noncarcinogenic effects. No data were available from which to determine
NOAELs, LOAELs,  and FELs for systemic effects (acute and chronic) and
reproductive toxicity by the oral route and for systemic effects
(chronic) by inhalation. No data were available for any parameters of
toxicity induced by dermal exposure.

2.3.2.3  Summary of relevant ongoing research

     The following information regarding ongoing research on benzene
toxicity was taken from the DIALOG (SSIE Current Research or Federal
Research in Progress) 1987 printouts:

     B. D. Goldstein (Rutgers Medical School) will study the role of
muconaldehyde, an intermediate in benzene metabolism, in benzene
toxicity. Three isomers of muconaldehyde will be synthesized; the
recovery of muconaldehyde from biological systems will be accomplished
using chemical and analytical techniques, the possible formation of
muconaldehyde in an in vitro mouse liver microsome system will be
studied, and the toxicity of muconaldehyde will be evaluated.

     0. A. Meyer et al. (National Institute of Environmental Health
Sciences) will identify methods sensitive to the neurobehavioral effects
of exposure to benzene during development and will attempt to
characterize any long-term neurobehavioral deficits following exposure
of rats to benzene during the postnatal period of development.

     M. T. O'Berg (E. I. du Pont de Nemours & Company) is conducting a
retrospective cohort study of 2,000 workers exposed to benzene between

-------
26   Section 2

1910 and 1976. Mortality of the cohort will be compared with expectation
based on the U.S. population as well as the entire work force of the Du
Pont Company, with particular attention to cancer, especially leukemia.

     Another epidemiological study of benzene-exposed workers, performed
by N. K. Weaver et al. (American Petroleum Institute), is in progress.
The study focuses on workers in petroleum operations. If primary studies
reveal excess mortality from specific diseases,  especially leukemia,
further studies will be performed to determine latency periods,  dose
relationships, and host factors.

     S. Lamm (Tabershaw Occupational Medicine Associates) is conducting
a historical epidemiological mortality study on chemical workers
involved in the production and use of benzene within the last 30 years
to measure the risk of illness or death in the exposed,  compared with an
unexposed, population.

     R. Snyder et al. (Thomas Jefferson University) are currently
studying mechanisms of benzene toxicity [i.e., benzene metabolism
(disposition) in three strains of mice that have exhibited different
sensitivities to the bone marrow depressant effects of benzene,  the
mechanism by which benzene inhibits cell replication in the regenerating
rat liver, and the effects of benzene and its metabolites on bone marrow
cells in culture].  R. Snyder is studying the effect of benzene on the
production of cell growth factors (hematopoietic factors).

     G. F. Kalf (Thomas Jefferson University) is using an in vitro
liquid culture system to study hematopoiesis and, in particular, the
ability of the stromal microenvironment to support hematopoiesis
following exposure to benzene. From these studies the investigator may
determine whether the site of benzene toxicity is the marrow adherent
layer, whether the target cell in the adherent layer is the macrophage,
whether toxicity results from the inhibition of growth factor production,
and whether hydroquinone and p-benzoquinone represent the toxic species.

     D. E. Nerland and H. E. Hurst (University of Louisville School of
Medicine) will study the effects of phenolic metabolites of benzene,
such as resorcinol, hydroquinone, and quinol. on erythropoiesis; the
immediate precursor of catechol, benzene dihydrodiol, will be tested for
bone marrow toxicity and certain pharmacokinetic properties.

     D. E. Nerland will also examine benzene metabolism and toxicity in
mice, rats, and guinea pigs exposed to low levels of benzene. Alpha and
beta interferon titers will be measured in mice to help determine
whether immunotoxicity is expressed prior to hemotoxicity.

     S. W. Burchiel (University of New Mexico) will use flow cytometry
and immunofluorescence to study the effects of several chemicals,
including benzene,  on murine bone marrow, spleen, and peripheral blood
cells, particularly as these parameters apply to immunotoxicity.

     A. C. Upton (New York University Medical Center) is conducting
studies which include development of an animal model for the
carcinogenesis of inhaled benzene or benzene in combination with known
leukemogenic agents in rats and mice.

-------
                                             Health Effects Summary   27

     R. Miday et al. (EPA, Cincinnati) are evaluating potential adverse
health effects in 35 persons exposed to well water contaminated with
benzene (8 ppm).

     N. S. Legator  (University of Texas Medical Branch at Galveston) is
conducting dominant lethal tests and cytogenetic analyses on mice
exposed to benzene by topical application or intramuscular injection

     The following  information regarding ongoing research on benzene
toxicity was obtained from Life Systems, Inc.:

     C. A. Snyder (New York University) is investigating the hematotoxic
effects of inhaled benzene at concentrations encountered in actual
environmental settings. Low-level exposures and sensitive hematopoietic
cell assays will be used to explore: (1) the nature of the dose and
temporal responses of low benzene concentrations. (2) whether precursor
cell toxicity observed at low concentrations foreshadows more extensive
hematopoietic damage, (3) the extent to which the toxic effects produced
at low concentrations of benzene are reversible, and (4) whether benzene
toxicity will be more severe if the hematopoietic system is given
additional stress.

     S. W. Burcheil (University of New Mexico)  is conducting studies to
examine the effects of benzene and other toxic substances on murine,
bone marrow, spleen, and peripheral blood cells using computer-based
multiparameter flow cytometry and immunofluorescence. This will provide
significant insights into the differential effects of benzene on immune
systems.

     D. Uierda (West Virginia University) is investigating the cellular
mechanisms of toxicity of benzene and benzene metabolites and bone
marrow immuno/hematopoiesis. The polyhydroxy metabolites of benzene to
be studied include benzoquinone, benzenetriol,  catechol, hydroquinone,
and phenol. A primary objective is to examine the relationship between
the in vivo and in vitro effects of benzene metabolites on bone marrow
precursor cell proliferation and differentiation.
     The following  information regarding ongoing research in the area of
benzene carcinogenicity was obtained from Dynamac Corporation:

     The Chemical Industry Institute for Toxicology  (CUT) indicated in
their 1986 annual report that a joint American Petroleum Institute/CIIT
research program has been initiated to study the mechanisms of benzene-
induced leukemia in mice. Although chronic exposure  to 100 ppm benzene
is believed to result in bone marrow damage and  increased incidence of
leukemia in man, the significance of exposures to low levels of benzene
is not known. This  study may help to clarify the relevance of the mouse
model in the assessment of human risk and may provide sensitive end
points for use as biological monitors of excess human exposure to
benzene.

     The benzene metabolite hydroquinone is being tested for
carcinogenicity. The compound has been administered  by gavage to rats
and mice in a 2-year NTP bioassay; currently the study is undergoing
quality assessment.

-------
 28    Section 2

      In addition to the  above,  other  ongoing research projects  on
 benzene have been reported for  this profile: E. P. Cronkite  (Brookhaven
 National Laboratory)  is  studying benzene  leukemocarcinogenesis  in mice,
 and T.  M.  Fliedner and Seidl  (University  of Ulm, Federal Republic of
 Germany) are studying carcinogenesis  and  other effects of  inhaled
 benzene in mice,  as well as tissue distribution and excretion.

 2.3.3  Other Information Needed for Human Health Assessment

 2.3.3.1  Pharmacokineti.es and mechanisms  of action

      The proposed mechanisms of action for human and animal
 toxicity/carcinogenicity are not fully understood on either  the cellular
 or  molecular level; however, the subject  has been studied extensively,
 and several theories-  have been  postulated.

      The database concerning absorption,  distribution, metabolism, and
 excretion of benzene  in  laboratory animals is extensive. Human data are
 more  limited,  particularly for  oral absorption, tissue distribution, and
 metabolism.  Additional information would  be helpful in clarifying the
 metabolism of benzene in bone marrow, as  would pharmacokinetic data
 concerning the binding of benzene metabolites to various tissues (i.e.,
 how long are the  metabolites bound to tissues?).

      The following information  regarding  ongoing research on benzene
 pharmacokinetics  was  taken from a computer printout from the DIALOG
 (SSIE)  (1987)  database:

      The percutaneous absorption of benzene will be assessed by I. H.
 Blank et al.  (Massachusetts General Hospital) using human cadaver
 stratum corneum in vitro,  human skin  in vivo, and monkey skin in vivo
 and in  vitro.

      R.  E.  Billings (SRI  International) is studying the hepatic
 formation  and toxicity of catechol metabolites of aromatic compounds,
 including  benzene.  The objective of the project is to (1) identify and
 characterize  the  specific enzymes involved in catechol formation and
 (2) examine  the mechanism of catechol formation and toxicity in isolated
 hepatocytes,  in perfused  liver,  and in vivo in rabbits and mice.

      The following information  regarding  ongoing research on benzene
 pharmacokinetics  was  taken from a toxicological profile on benzene by
 Life  Systems,  Inc., whose sources were the TOXLINE and CRISP databases
 and the  National  Institutes of  Health:

      R.  E.  Peterson (University of Wisconsin) has proposed a study of
 the prediction of metabolic pathways for benzene and other toxic
 chemicals  in rats  and mice  relative to the fact that there is an
 inhibition of the  metabolism related to the mixed-function oxidase
 enzyme  system.

      R.  Snyder (Rutgers,   State  University of New Jersey) is
 investigating the  relationship  between the production of metabolites of
benzene  and  the epidemiology of benzene - induced bone marrow depression
 and leukemia  in mice, rats, and rabbits.  The complete pathway of benzene
metabolism will be  investigated.

-------
                                             Health Effaces Summary   29

     V. F. Thomas  (University of Miami) will study the kinetics of
uptake, distribution, and elimination of benzene and other toxic
chemicals in man and in rats. Mathematical modeling will be used as a
tool.

     In addition to the above, it has been reported that N. Abraham (New
York Medical College) is studying the metabolism of benzene.

2.3.3.2  Monitoring of human biological samples

     Analytical methodologies exist for monitoring benzene levels in
expired breath and blood (see Sect. 8, Analytical Methods). These
techniques, particularly gas chromatography/mass spectrometry (GC/MS)
and gas chromatography/photoionization detection (GC/PID), have limits
of sensitivity in the sub-parts-per-billion range.  However, to date,
little information is available to correlate these monitoring end points
with NOAELs or LOAELs for specific biological effects.
     Urinary phenol levels have also been used as an index of benzene
exposure, particularly for occupational monitoring. At benzene levels of
5 ppm and higher, there appears to be a linear correlation between
exposure and phenol level; however, at lower exposures this correlation
can be obscured by potentially high background levels of urinary phenol
in unexposed individuals. Urinary phenol levels have not been used to
identify minimal adverse effect levels.

     Biological end points can be used to monitor benzene exposures.
Blood cell counts have been used as an indicator of high occupational
exposures, but not for NOAEL or LOAEL determinations. Another biological
end point that has been suggested is chromosomal aberrations in
lymphocytes; however, quantitative data are lacking.
     The following information on ongoing research on biological
monitoring for benzene exposure was obtained from searches of the SSIE
Current Research Data Base' and the Federal Research in Progress Data
Base on the DIALOG system.

     W. B. Coates, N. K. Weaver, and C. R. Stack (Hazelton Laboratory,
Vienna, Virginia) are conducting 5-day acute inhalation studies on rats
and mice to validate clinical procedures to be used in later long-term
studies. The current study focuses primarily on changes in blood
chemistry that may be early indicators of benzene-induced toxicity.

     D. E. Nerland (University of Louisville, Kentucky) is conducting
experiments on mice to determine whether benzene is immunotoxic.
Alpha/beta and gamma interferon titers will be quantified in animals
exposed to low concentrations of benzene. Quantification of interferon
production may provide a rapid method of screening workers exposed  to
benzene.

     G. A. Ansari  (University of Texas, Galveston) is conducting studies
on rats and in vitro studies with human plasma to determine whether
plasma proteins (in terms of their biological activity, concentration,
and covalent modification) can be used as markers of chemical exposure.
Benzene is one of the model compounds being used in the study.

-------
30   Section 2

2.3.3.3  Environmental considerations

     Analytical methodology.  The analytical methodologies of gas
chromatography (GC) coupled with flame ionization detection (FID),
photoionization detection (PID), or mass spectrometry (MS) provide for a
sufficient level of sensitivity (0.1 ppb and below) in measuring
environmental concentrations of benzene. GC/MS provides the greatest
Level of specificity but is generally not as sensitive as GC/FID or GC/PID.

     Bioavailability from environmental media.  The database concerning
che bioavaliability of benzene from food and environmental media is
good. Benzene levels have been documented in food,  air, soil,  and water,
and human uptake from all but soil is considered very possible.  Soil is
primarily an indirect source of benzene exposure via contaminated
groundwater.

     Environmental transport and fate.  The environmental transport and
fate of benzene are relatively well understood.  Additional information
concerning the leaching of benzene from various  soil types with varying
amounts of rainfall and differing microbial populations would, however,
be helpful.

     Interactions with other common cocontaminants.  Some studies have
been conducted on the interaction of benzene with other chemicals, both
in vivo and in the environment. Toluene, Arochlor-1254, phenobarbital,
and ethanol are known to alter the metabolism and toxicity of benzene.
These findings indicate that benzene metabolites, rather than benzene
itself, are the primary toxic agents in hematotoxicity and
immunotoxicity. Other than the reaction of benzene  with active
atmospheric species, such as ozone and hydroxy radicals, reactions with
nitrogen oxides and sulfur dioxide have been investigated. Undoubtedly,
although information concerning the reaction of  benzene with chemicals
other than those noted above does exist, it is unlikely that information
is available concerning the reaction with all the compounds with which
benzene will be found.

     Ongoing research.  No information was found concerning ongoing
research.

-------
                 3.  CHEMICAL AND PHYSICAL INFORMATION

3.1  CHEMICAL IDENTITY

     The chemical formula, structure, synonyms, and identification
numbers for benzene are listed in Table 3.1.

3.2  PHYSICAL AND CHEMICAL PROPERTIES

     The most important physical and chemical properties of benzene are
given in Table 3.2.

-------
32
Section 3
                                TiMeJ.l.  Chemical identity of benzene
                                                    Value
                                                                              References
   Chemical name

   Synonyms
   Trade name

   Chemical formula

   Wiswesser line notation


   Chemical structure
                                Benzene

                                Annulene. benzeen. benzen. benzm.
                                benzine, benzol, benzole, bicarburet
                                of hydrogen, carbon oil, coal naphtha.
                                cyclohexatnene, fenzen. mineral
                                naphtha, motor benzol. NCI-C55276,
                                nitration benzene, phene, phenyl
                                hydride, pyrobenzol. pyrobenzole
                                Polysirean
                                RH
I Oth C I Chem  Abstr

RTECS 1986. Oiemlme
1987. Wmdholz et al
1976. HSDB 1987
                                                                 IARC 1982
                                                                 RTECS 1987. as reported
                                                                 in HSDB 1987
                                              c=cx
                                         H-C      C-H
                                             \x      '/
                                              p-s
                                             H      H
  Identification numbers

    CAS Registry No                    71-43-2
    NIOSH RTECS No                  CY-1400000
    EPA RCRA Hazardous Waste No.      UOI9
    OHM-TADS No                     7216601
    DOT/UN/NA/IMCO Shipping No     Benzene 1114
    STCCNo.                          4908110
    Hazardous Substances Data Bank No.   35
                                                                10th C I Chem Abstr
                                                                Tatken and Lewis 1983
                                                                HSDB 1987
                                                                HSDB 1987
                                                                HSDB 1987
                                                                HSDB 1987
                                                                HSDB 1987

-------
                 Chemical  and  Physical  Information    33
Table 3.2. Physical and chemical properties of
Property
Molecular weight
Physical state
Odor
Odor threshold
Taste threshold
Melting point
Boiling point
Density"
Conversion factors
Solubility
Water
Nonaqueous solvents
Alcohol
Ether
Chloroform
Carbon disulflde
Acetone
Oils
Partition coefficient (log P)
(octanol/water)
Partition coefficient (log P)
(blood/air)
Vapor pressure

Viscosity
Heat capacity
Surface tension
Liquid-water interfacial tension

Latent heat of vaporization
Ratio of specific heats of vapor
Critical temperature
Critical pressure
Critical density
Soil adsorption coefficient (AToc)
Refractive index (nD:o)
Value
78 11
Clear colorless liquid
Rhombic prisms
Aromatic
4 68 ppm
05-45 mg/L
55'C
80 1"C at 760 mm Hg
0 8765 g/cm3
1 ppm — 3 26 mg/m3 at 20° C
1 mg/m3 — 0 31 ppm

0072%(wt/wt)at22°C
820 mg/L at 22°C
1000 mg/L at 25°C
1787 mg/L at 25°C
Miscible
Miscible
Miscible
Miscible
Miscible
Miscible
1 56-2.15
78
45 53 torr at IO°C
95 18 torr at 25°C
182.8 torr at 40.0°C
0125atmat 25°C
0.654 centipoue at 20° C
0 2499 cal/g/°C at 25°C
28 9 dyn/cm at 20° C
(0.0289 newton/m)
35 0 dyn/cm at 20°C
(00289 newton/m)
169 BTU/Ib
1061
289 5*C
48 7 atm
0 304 g/mL
0 3-100 (see Sect 22)
1 5011
References
Weast et al 1985
Windholz et al 1983
Weast et al. 1985
NFPA 1986
Weiss 1980
EPA 1975. as reported in
HSDB 1987
Weast et al 1985
Weast etal 1985
Weast etal 1985
Verschueren 1983

Jackman 1975
Chiou et al. 1977
Krasnoshchekova and Gubergnts
1975, as reported in CHEMFATE 1987
Chiou et al. 1982
Windholz et al. 1983
Windholz et al. 1983
Windholz et al. 1983
Windholz et al. 1983
Windholz et al. 1983
Windholz et al. 1983
Leo etal. 1971
Sato and Nakajuna 1979
Zwolinski and Wilhoit 1971. as
reported in CHEMFATE 1987
Thibodeaiu 1981
Jackman 1975
Jackman 1975
Weiss 1980
Weiss 1980
Weiss 1980
Weiss 1980
Jackman 1975
Jackman 1973
Jackman 1975
Rogers et al. 1980
Weast et al. 1985

-------
34     Section 3
                                            Table 3.2 (continued)
Property
UV absorption coefficient
(in water)
Sadtler reference number
Flash point
Autoignition temperature
Flammable limits
Burning rate
Heat of combustion
Vapor-air density
Vapor volume
Evaporation rate
Evaporation halflife
(from water)
Henry's law constant
Reactivity in water
OH Radical - Rate constant
Value
7000 L/mol-cm at 200 nm
220 L/mol-cm at 255 nm
6402 (IR. prism)
1 36 (IR. grating)
1765 (UV)
3429 (NMR)
I2°F
1044-F
1 3% (lower limit)
7 1% (upper limit)
6 0 mm/mm
-17.406 Btu/lb
1 4 at 100-F
37ft3
( 1 gal evapor )
2 8 (ether - 1 )
27h
3-5 h
5.5 X I0~3 atm m3/mol

31 X I010 L/mol-s
References
Setzlcorn and Huddleston 1965
Weast et al 1979, as
reported in HSDB 1987
NF7A 1986
NFPA 1986
NFPA 1986
Weiss 1980
Weiss 1980
NFPA 1986
AAI 1980
AAI 1980
Thomas 1982
Mackay and Leinonen 1975.
Mackay and Yeun 1983
Mackay and Leinonen 1975

Anbar and Neta 1967
                  Halfhfe
  Photodegradation in air

  Reactivity in air
    OH Radical  •  Rate constant
                 Halflife
    O3 Radical -  Rate constant
                 Halflife
                 Rate constant
                 Halflife
    O(3P) Radical • Rate constant
                                  078 X  1010 L/mol-s (25°C)
0 71 year
No direct photolysis; does not
absorb at 290 nm or longer
0.8 X 10~l2cm3/mcul-s(27eC)
0 13 X 10~" cm3/mcul-s (25'C)
0.114 X I0~" cm3/meul-s (2S°C)
24 5 days
0.7 X 10~22 cm3/mcul.i (24"C)
ca. 90 yean
0.7 X 10~22cm3/mcul-t(24aC)
126 yean
0.20 X I0~13 cm3/mcul-s (27"C)
Dorfman and Adams 1973, as
reported in CHEMFATE 1987
Anbar and Neta 1967
Howard and Durkin 1975. as
reported in CHEMFATE 1987
Cox et al. 1980
Gaffney and Levine 1979
Lorcaz and  Zellner 1983a
Coxctal. 1980
Pitts et al. 1979

Pate et al. 1976
                                                                    Hampton 1980, as
                                                                    reported in CHEMFATE 1987
      " Relative density Ratio of absolute density of benzene at 20°C to absolute density of water at 4°C.
       Data originally reported as cm /raol-s.

-------
                                                                      35
                         4.  TOXICOLOGICAL DATA
4.1  OVERVIEW
     Benzene is volatile and lipid-soluble and can be absorbed into che
body following ingestion, inhalation, and dermal contact.  Available data
from both animal and human studies indicate that after absorption
benzene must undergo metabolic transformation to exert its toxic
effects. Metabolism of benzene occurs primarily in the liver;  however,
the enzymes necessary for metabolism are also present in the bone
marrow, the putative organ of toxicity. The lymphoid system is another
target organ of benzene toxicity. It has been demonstrated
experimentally that the benzene metabolites benzene oxide, hydroquinone,
phenol, catechol, and trans,trans-mucondialdehyde can produce
hematotoxic effects. The metabolites hydroquinone, p-benzoquinone,
phenol, and catechol are known to cause lymphoid suppression.

     In humans, the hematotoxicity of benzene is characterized by
pancytopenia (a decrease in various circulating blood cells),  a
condition that reflects hypoplasia of the bone marrow. Some individuals
surviving bone marrow depression have developed myelogenous leukemia.
Benzene may also induce immunosuppression or sensitization.
     In addition to hematotoxicity and immunotoxicity, benzene can cause
neurotoxic effects  (drowsiness, dizziness, headache, vertigo,  delirium.
and loss of consciousness). Animal studies indicate that benzene is noc
teratogenic, but it has caused increased incidences of resorptions,
reduced fetal weight, skeletal variations, and altered fetal
hematopoiesis. Benzene is genotoxic, causing structural and numeric
chromosome aberrations, SCEs, and induction of micronuclei; however, it
has rarely been shown to cause gene mutations. The carcinogenicity of
benzene has been demonstrated in rats and mice. Epidemiological studies
suggest that long-term low-level exposure to benzene  is carcinogenic in
humans. Based on such evidence, EPA and IARC have classified benzene as
a human carcinogen  (leukemogen).

4.2  TOZICOKINETICS

4.2.1  Overview
     Following absorption into the body through  ingestion,  inhalation,
or dermal contact,  benzene  is widely distributed  to  tissues, with  the
relative uptake dependent on  the perfusion rate of  the tissues  by  blood
For example, accumulation in  fat  is  slow because  of  low perfusion, buc
the total potential uptake  is high  in  these  tissues because of  the high
lipid  solubility of benzene.

-------
 36    Section 4

      Metabolism of benzene  occurs  primarily  in  the  liver, where  it  is
 converted to benzene  oxide,  an unstable  intermediate which  forms phenol.
 There is  also evidence  that phenol is  formed from benzene by a direct
 insertion mechanism without proceeding through  a benzene oxide
 intermediate. Other metabolites include  catechol, hydroquinone, and
 conjugated phenolic compounds.  The primary oxidation of benzene  is
 catalyzed by enzymes  of the cytochrome oxidase  system, which occur
 primarily in the liver  but  are also found in bone marrow.

      Benzene is  excreted both  unchanged  via the lungs and as metabolites
 in  the urine. Phenol  and its conjugated  sulfates and glucuronides are
 excreted  in the  urine,  with unconjugated phenol the major urinary
 metabolite.  The  rate  and percentage of excretion via the lungs are
 dependent on exposure dose  and route.

 4.2.2  Absorpt ion

 4.2.2.1   Inhalation

     Human.   Data regarding the  inhalation absorption of benzene by
 humans consistently suggest  a  lung absorption factor of about 50% for
 continuous  doses  of 50  to 100  ppm  for  several hours (IARC 1982;  Nomiyama
 and Nomiyama 1974a,b; Sato  and Nakajima  1979; R. Snyder et al.  1981;
 Hunter 1968;  Srbova et  al.  1950).  Nomiyama and Nomiyama (1974a,b)
 exposed men and women to  benzene at 52 to 62 ppm for 4 h and estimated
 respiratory retention and uptake as  a  function of time during the
 exposure  period.  Respiratory uptake  (the difference between the
 concentration of  benzene  in  inhaled and  exhaled air expressed as
 percentage  of the concentration  in inhaled air) was measured at 47%,
 with little  difference  between men and women. Respiratory retention (the
 difference between respiratory uptake and excretion) was estimated at
 30% of the  inhaled dose.  Absorption was  greatest in the first 5 min and
 reached a constant  level  somewhere  between 15 min (Srbova et al.  1950)
 and 3 h (Nomiyama and Nomiyama  1974a,b) of continuous exposure.  Complete
 saturation  of body  tissues and  fluids may require several days (Gerarde
 1963).

     Animal.  Inhalation  studies with laboratory animals cor:inn that
benzene is  rapidly  absorbed  through  the  lungs,  although uptake is not
complete. Schrenk et  al.  (1941)  found that the initial absorption of
benzene in dogs was nearly complete  within 30 min,  reaching an
equilibrium within  several hours,  and that a linear relationship existed
between the concentration in air and the equilibrium concentration in
blood.

     An inhalation  study  with  rats  and mice recently presented at the
Society of Toxicology meeting  (Sabourin et al.  1986) indicated that
uptake is inversely proportional to  the exposure concentration.  The
percentage of inhaled benzene  that was absorbed and retained during a
6-h exposure period decreased  from  33% to 15% in rats and from 50% to
10% in mice as the exposure  concentration was increased from about 10 to
1,000 ppm.

-------
                                                 Toxicologies! Data   37

4.2.2.2  Oral

     Human.  Although definitive scientific data are not available on
oral absorption of benzene, case studies of accidental or intentional
poisoning indicate that benzene is readily and rapidly absorbed by the
oral route. Estimated oral doses ranging from 9 to 30 g have been fatal
in humans (Sandmeyer 1981). In addition, information gained from animal
studies (Parke and Williams 1953, Sabourin et al. 1986) indicates that
humans would absorb benzene with relatively high efficiency.
     Animal.  Benzene appears to be efficiently absorbed following oral
dosing. Parke and Williams (1953) were among the first to demonstrate
oral absorption of benzene. After ^C-labeled benzene was administered
orally to rabbits (0.34 to 0.5 g/kg), the total radioactivity eliminated
in exhaled air and urine accounted for -90% of the administered dose.
Recent data from the NTP (Sabourin et al. 1986) also indicate that
virtually all of an oral dose of benzene (0.5 to 150 mg/kg) is absorbed
by rats and mice.

4.2.2.3  Dermal

     Human.  Benzene can be absorbed through the skin, but the rate of
absorption is generally lower than that for inhalation exposure. No
studies were located regarding the cutaneous absorption of benzene
vapor. A recent investigation on the in vitro permeability of human skin
indicated that exposure of excised skin to benzene resulted in the
absorption of 0.17 mg/cm2 after 0.5 h and 1.92 mg/cm2 after 13.5 h
(Loden 1986). The initial absorption was in good agreement with an in
vivo study by Hanke et al. (1961), which showed that complete saturation
of a human forearm with benzene results in an hourly absorption of
0.4 mg/cm2. This rate of absorption was equal to 2% and 2 to 3% of that
of ethylbenzene and toluene, respectively (Dutkiewicz and Tyras 1967,
1968).

     Franz (1984) demonstrated, in vivo, that 0.023 ± 0.022% of the
benzene applied to human skin was absorbed; the remainder of the applied
dose quickly volatilized and was lost to the atmosphere. In vitro, the
total absorption of applied benzene was 0.1% for human skin, with peak
absorption occurring at 15-40 min. Total absorption increased 10-100
times when progressively larger doses, which persisted on the skin for
up to 3 h, were applied. Thus, it appears that a major factor
controlling the percutaneous absorption of benzene is its contact time
with the skin.

     Blank and McAuliffe (1985) calculated that an adult working in
ambient air containing 10 ppm benzene would absorb 7.5 /*L/h from
inhalation and 1.5 pL/h from whole-body (2 m2) exposure. They also
estimated that 100 cm2 of smooth and bare skin in contact with gasoline
containing 5% benzene would absorb 7.0 jjL/h. Blank and McAuliffe (1985)
noted that, because of the complex fate of benzene in the body, it is
difficult to relate quantitatively the amount of benzene in the body or
excreta to the amount penetrating the skin. Because of the degree of
water solubility of benzene, these authors propose that diffusion
through the stratum corneum is the rate-limiting step for dermal
absorption.

-------
38   Section 4

     Based on skin absorption studies with mice (Susten et al. 1985),
NIOSH calculated  that a worker exposed to benzene as a result of skin
contact with petroleum naphtha, a solvent commonly used in tire
manufacturing, could absorb -6 mg of benzene through intact skin. This
amount absorbed was compared with an estimated 14 mg of benzene absorbed
as a result of inhalation of 1 ppm for an 8-h day. Absorption through
abraded skin was  estimated to be 5.3 times higher than through intact
skin (OSHA 1985). A similar calculation was performed by Johnson (1979,
as reported in Susten et al. 1985), based on data from a study by
Maibach and Anjo  (1981) in which a rubber solvent containing 0.35%
benzene was applied repeatedly to the forearms of rhesus monkeys. From
these data, Johnson calculated that -0.4 to 0.9 mg of benzene would be
absorbed through  the skin of rubber workers with one palm in contact
with the solvent  30 times a day. If Johnson had based his estimates on
contact with two  palms 150 times per day as did Susten et al. (1985), 4
to 9 mg per day could be absorbed (compared to 6 mg per day calculated
by Susten et al.).

     Animal.  In  rhesus monkeys and hairless mice, dermal absorption was
<1% following a single direct application of liquid benzene (Maibach and
Anjo 1981, Susten et al. 1985). Multiple applications as well as
application to stripped skin resulted in greater skin penetration
(Maibach and Anjo 1981). For the monkey and mini-pig, total in vivo
dermal absorptions were 0.14 and 0.09%, respectively, of the applied
dose; total in vitro absorptions were 0.19 and 0.23%, respectively
(Franz 1984).  The total absorption of benzene in each of these animals
was approximately twofold higher than that of man.

4.2.3  Distribution

4.2.3.1  Inhalation

     Human.  An early study indicates that humans exposed to high doses
of benzene (6,000 ppm) absorb -30% into the blood; over half of the
absorbed portion  is translocated into bone marrow, adipose tissue, and
liver (Ouvoir et  al. 1946). Benzene is relatively insoluble in body
fluids;  however, because of its high lipid solubility, it may be stored
and accumulated in fatty tissues (Sandmeyer 1981). Sato et al. (1975)
showed that the amount of body fat influences the toxicokinetics of
benzene.  In experimental human inhalation exposures, the benzene
concentration in blood was consistently lower in females than in males.
Elimination was slower in females and attributed primarily to the
relatively higher fat content (Sato et al. 1975).

     Tissue levels of benzene have been reported in cases of both
accidental and intentional exposure.  Tauber (1970) reported levels of
0.38 mg% in blood (mg% - mg per 100 mL of blood or mg per 100 g of
tissue),  1.38 mg% in the brain, and 0.26 mg% in the liver of a worker
who died from exposure to high air concentrations of the chemical.
Autopsies from individuals who died after "sniffing" different materials
containing benzene revealed a range of concentrations in various
tissues:  blood, 0.094 to 6.4 mg%; brain, 1.38 to 3.9 mg%; liver, 0.26 to
1.6 mg%;  kidney,  0.55 mg%; urine 0.06 mg%; bile, 1.1 mg%; fat, 2.23 mg%;
and stomach, 1.0  mg% (Winek et al. 1967, Uinek and Collom 1971).

-------
                                                 Toxicological Data   39

     Animal.  A series of inhalation studies conducted with dogs
(Schrenk et al. 1941) indicate that benzene is rapidly distributed
throughout the body. Fat, bone marrow, and urine contained about 20
times the concentration of benzene in blood; benzene levels in muscles
and organs were one to three times that in blood; and RBCs contained
about twice the amount of benzene found in plasma.
     Following inhalation exposure of rats to 500 ppm, benzene levels
reached a steady-state concentration within 6 h in blood (11.5 Mg/g),
bone marrow (37.7 j*g/g) ,  and fat (164.4 /*g/g).  Lower concentrations were
found in kidney, lung, liver, brain, and spleen. Phenol, catechol,  and
hydroquinone were detected in blood and bone marrow following 6 h of
exposure to benzene, with levels in bone marrow exceeding the respective
levels in blood. The levels of phenol in blood and bone marrow decreased
much more rapidly after exposure ceased than did those of catechol or
hydroquinone, suggesting the possibility of accumulation of the latter
two compounds (Rickert et al.  1979).

     Sato et al. (1974) noted that benzene was stored longer and
eliminated more slowly in female and male rats with large body fat
content than in lean animals.
     The relative uptake in tissues appears to be dependent on the
perfusion rate of tissues by blood. Following a 10-min inhalation
exposure of pregnant mice, benzene was found to be present in lipid-rich
tissues, such as brain and fat, and in well-perfused tissues, such as
liver and kidney. Benzene was also found in the placenta and fetuses
immediately following inhalation of benzene (Ghantous and Danielsson
1986).

     C. A. Snyder (1987)  notes that bioaccumulation and rates of
elimination of benzene are expected to be different from those of its
metabolites, due to differences in physical and chemical properties.
Benzene is nonpolar, neutral, and lipid soluble, while the metabolites
are polar, acidic, and water soluble. Reflecting these differences are
studies which show that the bone marrow/blood gradient for benzene in
rats and mice is 3 to 4 (Rickert et al. 1979; C. A. Snyder et al. 1977,
as reported in C.A. Snyder 1987), while that of phenol is about 0.3
(Greenlee et al. 1981). The bone marrow/blood gradient for catechol and
hydroquinone, two other benzene metabolites, is about three times
smaller than that for benzene (Greenlee et al. 1981).

4.2.3.2  Other routes of exposure
     Distribution studies in humans and animals were not found for the
oral or dermal routes of exposure.
     One study suggests that benzene crosses the human placenta, with
levels in cord blood equal to or greater than those measured in maternal
blood (Dowty et al. 1976).

4.2.4  Metabolism
     Many of the studies elucidating the biotransformation of benzene
used routes of exposure other than  inhalation, oral, and dermal. There
is no available evidence to suggest  that the route of administration has
any substantial effect on the subsequent metabolism of benzene, either

-------
 40   Section A

 in humans or in animals. For these reasons,  as well as for clarity of
 the discussion, this section will not be divided into route-specific
 sections.

      Data regarding human metabolism of benzene have been limited to
 studies that identify unmetabolized benzene  in the breath and phenolic
 metabolites in the urine of humans exposed by inhalation and are
 discussed in the following section on excretion.  Qualitatively,  the
 metabolism and elimination of benzene appear to be similar in humans and
 laboratory animals, but no directly comparable studies are available.

      Early metabolic studies with experimental animals focused on the
 identification and quantitation of metabolites,  while more recent
 investigations have emphasized the mechanism of the initial benzene
 oxidation and have characterized the reactive species responsible for
 benzene-induced toxicity.  The liver is the major  site of benzene
 metabolism;  consequently much of the research has focused on this organ.
 Benzene metabolism has been studied in vitro using liver homogenates,
 cell supernatant fractions containing microsomes,  and preparations of
 microsomes.  Most of the available evidence suggests that benzene
 toxicity is  produced by one or more metabolites rather than by benzene
 itself;  however,  bioactivation of benzene  is complex.

      The metabolism of benzene initially involves  its oxidation  to form
 hydroxylated benzenes,  with phenol as the  major metabolite.  The  enzymes
 catalyzing the  hydroxylation of benzene are  mixed-function oxidases
 which are  mostly found in  the liver,  but also occur in bone marrow,  the
 target organ of benzene toxicity (C.  A.  Snyder 1987).  Possible routes of
 biotransformation are shown in Fig.  4.1.

      C.  A. Snyder (1987) proposes that the formation of the major
 hydroxylated benzenes involves  its oxidation via  two concurrent
 pathways,  one by  direct hydroxylation and  the other by indirect
 hydroxylation via an epoxide  (benzene oxide)  intermediate.  The direct
 oxidation  of benzene is thought to be mediated by one  or more  free
 radicals,  superoxide anion,  or  hydrogen peroxide. In addition, an enone
 (cyclohexadione)  may be involved in both the  direct  and the  indirect
 hydroxylation of  benzene (Hinson et al.  1985).

      Further metabolic  products are  formed by the introduction of a
 second hydroxyl group to form hydroquinone and/or catechol  or  a  third
 hydroxyl group  to  form  1,2,4-trihydroxybenzene or by  conjugation
 reactions  to produce glucuronides and sulfate  esters before urinary
 excretion. Additionally, the  hydroxylated  metabolites  can be oxidized to
 their corresponding  quinones  or semiquinones  (C. A.  Snyder  1987).  In
 guinea pig microsomes,  hydroquinones  are products of phenol metabolism
 that may be  further  oxidized  to yield p- or o-benzoquinone  (Smart  and
 Zannoni 1985). A minor  metabolite,  phenylmercapturic  acid, may be  formed
by enzymatic reactions  of benzene oxide with  glutathione (Jerina  et al.
 1968).

     Because of the  stability of the benzene  ring, only a small number
of ring-opened metabolites are  formed. House  liver microsomes  catalyze
ring opening in the  presence  of NADPH, producing muconaldehyde, a known
hematotoxin  (Latriano et al.  1986). Small amounts of  trans,trans-muconic

-------
                                                  101
                                MDRECT
                                HVDROXVLATION
                       OLUTATHIONE
              CO2H     TRANSFERASE
              I
          SCHaCM

              I
          CH3CNH

             II
             O
-ALANME

-OlYCME
 PHENVLhCRCAPTURC ACD
                                                                 101
                                                                 DRECT
                                                                 HVDROXYLATON
                                                                   CONJUGATIONS
                                                                                    SULMTES ESTERS .
                                                                                    GLUCURONDES
                                                                                    ELMNATEO M URME
 /   I   \
DBECT • MOMECT
 HVDROXVLATION
                                        OH
                                                       /HVDROXVLATION   V



                                                            t       \
                                                                                                       OH
                              IOI
      C=C
H02C
                    AOD
                      8ULMTES.ESTERS.*
                      OLUCURONDES
                      ELMMATEO M URME
                                                                        REACTIONS WITH NUCLEOPHLIC
                                                                    MACROMOLECULES PROTENS DNA RNA
                                                                                                                                    n
                                                                                                                                    o
                                                                                                               to
                                                                                                               n
                                                                                                               to
                                        Fig. 4.1.  Bioiransformation of benzene.

-------
42   Seccion 4

acid (Che corresponding diacid of muconaldehyde) were found in the urine
of rabbits and mice receiving oral doses of ^-^C-labeled benzene (Parke
and Williams 1953, Gad-el-Karim et al. 1985). The expired air of rabbits
also contained minor amounts of labeled carbon dioxide,  indicating
complete benzene metabolism (Parke and Williams 1953).
                       •   '
     The liver, the repository of cytochrome enzymes thought to be
involved in the oxidation of benzene in mammalian systems, plays an
important role in the bioactivation of benzene. Sammett et al. (1979)
provided corroborative evidence for this role by showing that partial
hepatectomy of rats diminished both the rate of metabolism of benzene
and its toxicity, suggesting that a metabolite formed in the liver is
necessary for toxicity.

     Benzene metabolism in bone marrow is not clearly understood.
Although bone marrow possesses a limited capacity to metabolize benzene,
it is insufficient to account for the levels of metabolites (phenol,
hydroquinone, and catechol) in this tissue. The rate of benzene
metabolism in bone marrow is lower than that in liver and is attributed
to the low level of mixed-function oxidase activity found in bone
marrow. Phenol, however, appears to be readily metabolized by bone
marrow (Irons et al. 1980a, Irons 1985).

     A number of investigators have suggested that covalent binding of
benzene metabolites to cellular macromolecules is related to its
mechanism of toxicity. For example, benzene metabolites have been found
to bind to proteins in mouse liver, bone marrow, kidney, spleen, blood,
and muscle (Longacre et al. 1981a); to proteins in perfused bone marrow
preparations (Irons et al. 1980a) and in rat liver DNA (Lutz and
Schlatter 1977);  and to DNA in rabbit and cat bone marrow mitochondria
(Rushmore et al.  1984). The inhibition of RNA synthesis in liver and
bone marrow mitochondria has been correlated with covalent binding of
benzene metabolites to DNA (Kalf et al. 1987.)

     Benzene has been found to stimulate its own metabolism, thereby
increasing the rate of toxic metabolite formation. Pretreatment of mice
with benzene stimulated benzene metabolism by liver microsomes, while
pretreatment with phenolic metabolites did not (Dean 1978, Gonasun et
al.  1973).  The rate of benzene metabolism can be altered by pretreatment
with various compounds. Benzene is metabolized largely by mixed-function
oxidases in the hepatic microsomes. Therefore, chemicals which stimulate
the activity of this enzyme system also increase the rate of benzene
metabolism.  Phenobarbital pretreatment has been shown to increase the
rate of benzene metabolism by 40% in rats and 70% in mice. SKF-52SA
inhibited benzene metabolism in the rat,  while toluene inhibited benzene
metabolism in both rat and mouse (R. Snyder and Kocsis 1975, Sato and
Nakajima 1979). Gonasun et al. (1973) reported that carbon monoxide,
aniline,  aminopyrine,  and cytochrome C metapyrene inhibit benzene
metabolism in mouse liver microsomes. Ethanol ingestion as well as
dietary factors,  such as food deprivation and carbohydrate restriction,
also enhance the metabolism of benzene in rats (C.A. Snyder et al.
1981a,  Sato and Nakajima 1985).

-------
                                                 Toxi.coLogi.caL Data   M

4.2.5  Excretion

4.2.5.1  Inhalation

     Hunan.  Following inhalation exposure to benzene, humans eliminate
the compound in unchanged form in the exhaled air and in metabolized
form in urine. Estimates of the fraction of benzene excreted in the
expired air have ranged between 12 and 50% (Teisinger et al. 1952,
Srbova et al. 1950; Nomiyama and Nomiyama 1974a,b)  The respiratory
elimination is described as triphasic. The initial component has a  half-
life of -0.9 h, similar to the value determined in rats (Rickert ec al
1979).  The second, slower phase has a half-life of 3 h, and the third
has a half-life of >15 h. No differences in respiratory elimination were
observed between men and women (Nomiyama and Nomiyama 1974a,b).
     Hunter (1968) reported that the level of benzene in expired air
rapidly decreased when exposure ceased, but that benzene could be
detected up to 24 h after exposure, suggesting that it is possible  to
back-extrapolate to the concentration in inspired air. However, the
amount of benzene excreted in expired air will vary not only with the
dose, but also with the extent of metabolism in the body. Consequently,
levels in the breath may not be proportional to the dose.
     Urinary excretion of benzene metabolites, primarily phenol,
constitutes another important pathway for benzene elimination. Most of
the phenol is excreted in the form of sulfate esters (Teisinger et  al.
1952),  but significant amounts of glucuronides may be present,
especially after exposure to high concentrations of benzene (Sherwood
1972).  In an inhalation study conducted by Teisinger et al. (1952),
28.8% of the absorbed benzene was excreted as phenol, 2.9% as
pyrocatechol, and 1.1% as hydroquinone. The urinary excretion was
highest within the first 24 h following exposure and was essentially
complete within 48 h, while the urinary excretion of hydroquinone was
slower, but still usually completed within 48 h.
     Urinary phenol excretion has been used as an indicator of overall
benzene exposure, with urinary levels of 75 mg/L indicating an exposure
to about 10 ppm (8-h TWA) (NIOSH 1974) and 100 mg/L indicating an
exposure to 200 ppm-h (25 ppm for 8 h) (Sandmeyer 1981). Recently,  Inoue
et al.  (1986) measured the urinary excretion of phenol in workers
following a 7-h workshift exposure to 1 to 76.4 ppm benzene and obtained
a correlation of 0.891 between exposure level and urinary phenol
excretion.
     Animal.  As in humans, benzene is eliminated by expiration from  the
lungs and via excretion in the urine of experimental animals. Only a
small amount is excreted in feces. Exhalation is the major route of
excretion of unmetabolized benzene (IARC 1982). A biphasic pattern of
elimination was observed in rats exposed to 500 ppm for 6 h, with half-
times for expiration of 0.7 and 13.1 h (Rickert et al. 1979). The
initial half-life of 0.7 h was similar for blood, bone marrow, and other
organs; fat, with a half-time of 1.6 h, was the only tissue that
differed markedly from blood.
     The major route of excretion  following a 6-h inhalation exposure  of
rats and mice to various concentrations of ^-^C-benzene appeared  to be

-------
 44   Section 4

 dependent on the Inhaled concentration (Sabourin  et  al.  1986).  At  lower
 concentrations (13 to 130 ppm in rats  and 11  to 130  ppm  in mice),  >85%
 of the excreted radioactivity was identified  as urinary  metabolites  and
 <6% was in the expired air.  At higher  concentrations (260  or  870 ppm in
 rats and 990 ppm in mice),  11 to 48% of the excreted radioactivity was
 exhaled in the expired air as unmetabolized benzene,  suggesting
 saturation of the metabolic  pathways for benzene.

 4.2.5.2  Oral

      Human.   No studies are  available.

      Animal.   Parke and Williams (19S3)  administered radiolabeled
 benzene by oral intubation to rabbits  and recovered  43%  of the  label  as
 exhaled,  unmetabolized benzene and 1.5%  as carbon dioxide.  Urinary
 excretion amounted to about  35% of the  dose. The  isolated  urinary
 metabolites  were mainly in the form of phenolic sulfates and  glucuronides
 and included phenol (23%), hydroquinone  (4.8%), catechol (2.2%),
 trans,trans-muconic acid (1.3%),  phenylmercapturic acid  (0.5%), and
 1,2,4-trihydroxybenzene (0.3%).  The rest of the radioactivity (5 to  10%)
 remained in  the tissues or was excreted in the feces. This  same general
 profile of urinary metabolites is also  found in rats  (Cornish and  Ryan
 1965),  mice  (Longacre et al.  1981a), and cats and dogs (Oehme 1969).  the
 chemical  structures of the urinary metabolites are given in Fig. 4.2.

      The  effect of dose on the excretion of ^-^C-benzene  administered
 orally  has been studied in rats and mice (Sabourin et al.  1986). At
 doses of  <15  mg/kg,  >80% of  the administered dose was excreted  in  the
 urine of  both species.  As the dose was  Increased from 15 to 150 mg/kg.
 the  amount of benzene exhaled in the breath increased to 50%  of the
 administered  dose  in rats and 71% in mice. The exhaled benzene was
 largely in the  form of unmetabolized benzene,  again  suggesting  that
 saturation of the  metabolic  pathways had occurred.

 4.2.5.3   Dermal

     Human.   No  studies are  available.

     Animal.   No studies are available.

 4.3  TOXIC ITT.

 4.3.1   Lethality and Decreased Longevity

 4.3.1.1  Overview

      Individual case reports  of acute benzene lethalities have appeared
 in the  literature  since the  early 1900s.  The benzene concentrations
 encountered by  the  victims were  not often known.  However,  tissue levels
 of the chemical have been reported for some victims.  Lethality  in humans
has been tentatively attributed to  asphyxiation,  respiratory  arrest,
central nervous system (CNS)  depression,  or cardiac arrhythmia.

     Benzene  appears to be of low acute  toxicity when administered to
animals by various  routes, but sudden death may occur at high
concentrations. Lethality in animals has been ascribed to ventricular
fibrillation  or accelerans stimulation.

-------
                                         Toxicological Daca
   OH
           PHENOL
  OH
           HYDROQUINONE
  OH
  OH
       OH
          CATECHOL
H02C
      H

H      Nc-n'C02H
 \     xc-c\   *
                  H        trans. trana-MUCONIC ACID
         H
        S—CH0—CH —
                I
               HN
                         COgH
                                      PHENYLMERCAPTURIC ACID
                    —CH,
 OH
     OH
         1.2.4.-TRWYDROXYBEN2ENE
OH
            Fig. 4.2. Urinary metabolites of benzene.

-------
46   Section 4

4.3.1.2  Inhalation

     Human (case reports).  Tauber (1970) reported a case of acute
benzene toxicity in which the chemical overflowed from a tank in a light
oil loading area. Sudden death, attributed to the combination of high
air concentrations of benzene, excitement and running on the part of the
victim, and the presence of toluene in the atmosphere, occurred in a
worker exposed to the fumes. Benzene levels were 0.38 mg% in the blood,
1.38 mg% in the brain, and 0.26 mg% in the liver.

     Winek and CoHorn (1971) reported three cases of accidental death
due to intentional acute exposure to benzene. One boy, who died as a
result of "sniffing" rubber cement containing benzene, had a blood level
of benzene of 0.094 mg% and a kidney level of 0.55 mg% (Winek et al.
1967). Another death occurred when a boy sniffed straight benzene. In
this case, benzene tissue levels were: 2.0 mg% in the blood, 0.06 mg% in
the urine, 1.9 mg% in the kidney, 1.6 mg% in the liver, 1.1 mg% in the
bile, 2.23 mg% in fat, 3.9 mg% in the brain,  and 1.0 mg% in the stomach.
In a third victim, who was sniffing glue and accidentally shot himself
while in a euphoric state, the blood level was 6.5 mg%.

     At autopsy, Winek and Collom (1971) observed inflammation of the
respiratory tract, hemorrhages of the lungs,  congestion of the kidneys,
and cerebral edema, but no hematological effects, even when benzene
levels in the blood were as high as 2 mg/100 mL. The suggested causes of
death following exposure to high concentrations of benzene were
asphyxiation, respiratory arrest, CNS depression, or cardiac arrhythmia
(Winek and Collom 1971; R. Snyder and Kocsis 1975, as reported in
Andrews and Snyder 1986).

     Hamilton (1922) reported on several work-related cases of lethal
acute benzene exposures, some of which occurred in spite of extensive
precautions. In two cases, rescuers died.

     Based on the data of other investigators, Sandmeyer (1981)
correlated the signs and symptoms of acute benzene toxicity via
inhalation (the most common route of exposure) with concentration and
duration of exposure. Sandmeyer (1981) estimated that exposure to
benzene concentrations of 19,000 to 20,000 ppm for 5 to 10 min may be
fatal. The rapidity of death in such cases suggests that acute lethality
may be caused by benzene itself, not a metabolite.

     Animal.  Animal lethality data indicate that benzene is of low
acute toxicity via the respiratory route (0'Bryan and Ross 1986). An
inhalation LC50 value for rats was calculated as 13,700 ppm for a 4-h
exposure (Drew and Fouts 1974). Smyth et al.  (1962) reported that 4 of 6
rats died following a 4-h exposure to 16,000 ppm benzene.

     Nahum and Hoff (1934) investigated the mechanism of sudden death in
benzene toxicity. Cats and monkeys, having undergone various treatments,
were exposed to high concentrations of benzene. The intact animals
exhibited extrasystoles and ventricular tachycardia of a pre-
fibrillation type. Adrenalectomy reduced, but did not abolish, the
ventricular extrasystoles; removal of both stellate ganglia did not
reduce the frequency of ventricular rhythms.  However, the ventricular
rhythms were abolished by removal of both the adrenals and the ganglia
and were restored by injections of adrenalin. Respiratory failure also

-------
                                                  Toxicological  Data    47

 occurred during the period when the animals were narcotized.  The
 Investigators concluded that benzene causes the liberation of adrenalin
 and sensitizes the myocardium to its action and that death may  occur
 suddenly from ventricular fibrillation or accelerans stimulation  or
 both.  Other animals died from respiratory failure.

      In addition to the studies listed below,  other acute  lethality daca
 for benzene in experimental animals have been reviewed in  Sandmeyer
 (1981)  and IARC (1982).

 4.3.1.3  Oral

     Human.   The lethal oral dose  of benzene  in humans has  been
 estimated at  10 mL (8.8 g)  (Thienes and Haley 1972,  as reported in
 Sandmeyer 1981).

     Lethal oral doses  have produced signs  and symptoms of  staggering
 gait, vomiting,  shallow and rapid  pulse,  somnolence,  and loss of
 consciousness,  followed by  delirium,  pneumonitis, collapse, and then
 sudden  CNS  depression  (Von  Oettingen 1940,  as  reported in Sandmeyer
 1981);  more moderate doses  produce dizziness,  excitation, and pallor,
 followed  by flushing, breathlessness,  headache,  weakness, constriction
 of  the  chest,  and fear  of impending death (Sandmeyer  1981). The victims
 may experience  visual disturbances and convulsions.  Feelings of
 excitement and  euphoria may quite  suddenly  change to  weariness, fatigue
 and sleepiness,  followed by coma and death  (Lurie 1952, as  reported in
 Sandmeyer 1981).  In one  case,  accidental  ingestion  of benzene may also
 have induced  ulceration of  the gastrointestinal  mucosa (Appuhn  and
 Goldeck 1957; Caprotti  et al.  1962.  both  as reported  in EPA 1980b).

     Animal.  Animal lethality data indicate  that benzene is of low oral
 acute toxicity  (0'Bryan  and Ross 1986). Oral  LD50 values for rats ranged
 from 0.93 to  5.96  g/kg  (Cornish and Ryan  1965, as reported  in IARC 1982,
 Withey and Hall  1975);  the  values  varied  with  age and strain of the
 animals (Kimura  et  al.  1971, as reported  in Sandmeyer 1981). In the
 mouse the oral  LD5Q was  4.7  g/kg (Savchenko 1967, as  reported in
 Sandmeyer 1981).

 4.3.1.4  Dermal

     No information was  available  regarding benzene-induced lethality in
 humans or animals via dermal exposure.

 4.3.2  Systemic/Target Organ Toxicity

     The most significant health effects  of benzene are hematotoxicity.
 immunotoxicity, and neurotoxicity.

 4.3.2.1  Hematotozlc ity

     Overview.  Humans exposed to  benzene have developed marked
hypoplasia of the bone marrow  with pancytopenia  (a decrease in  the
various cells of the circulating blood);  some  individuals surviving the
marrow depression have developed myelogenous  leukemia.

     Pancytopenia and its variations and  aplastic anemia have been
detected following  chronic  exposure  to benzene in a variety of

-------
48   Section 4

situations, primarily occupational. The case studies reviewed below
characterize the variety of hematological parameters that can be altered
in humans exposed  to benzene.

     In one study, pancytopenia developed in 32 workers exposed for
periods ranging from 4 months to IS years to estimated concentrations of
ISO  to 650 ppm benzene. However, these and other data found in the
literature fail to establish a relationship between the extent of
exposure and effect. Only one study was found which attempted to
establish a correlation for noncarcinogenic hematotoxicity in workers
exposed to low concentrations of benzene, -20 ppm. From these data, the
investigator estimated that benzene poisoning may occur at levels of
about 10 ppm.

     The hematotoxic effects observed in humans have been reproduced in
animals. Although more animal than human data are available from which
to determine low- or no-effect levels of benzene hematotoxicity, the
data show that animal responses to benzene exposure are variable and may
depend on factors such as species (differences observed in various
species may be due to inherent interspecies variability and/or
differences in exposure regimens), strain, duration of exposure, or
whether exposure is intermittent or continuous (the same concentration
of benzene is more toxic when exposure is continuous).  Also, wide
variations have been observed in normal hematological parameters,
complicating statistical evaluation. However, in spite of the
variability, some consistent findings in benzene toxicity have been
noted. For example, lymphocyte levels have been depressed most severely
and in the shortest time, anemia does not occur as frequently as
lymphocytopenia, and granulocytes appear to be the most resistant of the
circulating cells in response to benzene exposure.

     The experimentally induced hematological effects of benzene appear
to be the same regardless of route of administration. Leukopenias have
been reported in animals exposed subchronically via inhalation to
concentrations of 60 or 88 ppm and via oral administration of doses of
SO mg/kg.  Dose responses have been demonstrated in several cases.

     Numerous studies have shown that benzene-induced bone marrow
depression is the result of damage to the pluripotential stem cells
and/or the early proliferating committed cells in either the erythroid
or the myeloid lines. Pluripotent stem cell levels have been altered by
short-tern exposure to 100 ppm benzene. Other studies have demonstrated
suppression of the UBC and R£C lines.

     There is general agreement among the various investigators in the
field of benzene coxicity that benzene metabolites, not benzene, are the
primary toxic agents in the induction of hemato- and immunotoxicity.
This agreement has evolved from studies in which agents known to alter
benzene metabolism (toluene, Aroclor-1254, phenobarbital, and ethanol)
have also altered benzene toxicity. Toluene, Aroclor-1254, and
phenobarbital appear to alleviate benzene toxicity, while ethanol
generally increases benzene toxicity.

     The hematotoxic effects of the benzene metabolites benzene oxide,
hydroquinone, phenol, catechol, and trans,trans-mucondialdehyde have
been demonstrated experimentally.

-------
                                                 lexicological Data   49

     Inhalation, human.  Aksoy et al.  (1972) studied 32 patients that
had been exposed to benzene concentrations estimated at 150 to 650 ppm
(based on spot measurements) for 4 months to 15 years. These individuals
exhibited severe blood dyscrasias that included pancytopenia with
hypoplastic, hyperplastic, or normoplastic bone marrow. Eight of the 32
patients died with thrombocytopenic hemorrhage and infection.

     Yin et al. (1987b) conducted a study in China involving 508,818
workers exposed to benzene. Of these workers, 4.98% were exposed to
benzene alone, and 95.02% were exposed to mixtures containing mainly
benzene, toluene, and xylene. The levels of benzene detected at 95% of
the work stations ranged from 0.06 to 844 mg/m3. One and one-third
percent of the work stations had levels of >1,000 mg/m3, and 64.6% had
levels of <40 mg/m3. In the overall study, 2,676 cases of "benzene
poisoning" were found (24 aplastic anemia, 9 leukemia, and the remainder
presumably leukopenia). The prevalence rate of "benzene poisoning" was
0.94% in workers exposed to benzene alone and 0.44% in workers exposed
to mixtures. One of the major findings of the study was in the
shoemaking industry. "Benzene poisoning" was found in 28 of the 141
shoemaking factories studied (124 cases in 2,740 employees). A positive
correlation was observed for prevalence of poisoning and benzene
concentration (correlation coefficient, 0.42; P < 0.05). In one
workshop,  there were four cases of aplastic anemia in 211 workers
(1.25%) exposed to benzene and chlorobutadiene (3:1) for 118.5 days to a
mean concentration of 1,035.6 mg/m3.  The prevalence of benzene-induced
aplastic anemia in the shoemaking industry was about 5.8 times that in
the general population.

     Findings similar to these have been reported in the literature for
other individuals or groups of individuals chronically exposed to
benzene in occupational situations.  For example:

   •  pancytopenia has been reported in one individual with 13 years of
     exposure in which bleeding was observed with a normal platelet
     count;  the platelets were, however, functionally and
     morphologically abnormal (Favre-Gilly and Bruel 1948, as reported
     in Goldstein 1977);

   •  23/332 rotogravure workers exposed for 6 to 60 months to benzene
     vapor concentrations of 11 to 1,069 ppm had severe hematological
     abnormalities (Goldwater 1941);

   •  6/217 apparently healthy workers in small shoe factories exposed to
     benzene concentrations of 30 to 210 ppm for 3 months to 17 years
     had pancytopenia (Aksoy et al.  1971, as reported in Goldstein
     1977);

   •  six rotogravure printers exposed to 24 to 1,060 ppm (Erf and Rhoads
     1939,  as reported in Goldstein 1977) had pancytopenia. It was
     suggested that bone marrow is hyperplastic early in toxicity and
     hypoplastic later on;

   •  1/216 workers exposed in the watch industry (Guberan and Kocher
     1971,  as reported in Goldstein 1977) died of aplastic anemia;

-------
 50    Section  4

    •  25/1,104 workers  in a rubber  factory exposed to concentrations of
      up  to  500  ppm (average,  100 ppm)  developed severe pancytopenia; 83
      others had mild hematological  abnormalities (Wilson 1942, as
      reported in Goldstein 1977).

      Other  cases are reviewed in Hamilton (1922), Aksoy et al  (1976)
 Goldstein  (1977),  and  IARC (1982).

      Inhalation,  animal.   Generally, hematotoxicity has not been
 associated  with acute  inhalation exposure to benzene. However, the
 following subacute and subchronic studies demonstrate adverse
 hematological effects  in animals, similar to those observed in humans.

      Li  et  al.  (1986)  examined the  effect of benzene and other solvent
 vapors on peripheral blood alkaline phosphatase levels in female Vistar
 rats. The animals  were exposed to 0, 20, 50, 100, 300, 1,000, or
 3,000 ppm benzene  for  7 days,  and enzyme activities were determined on
 serum and leukocytes.  Serum levels  of  the enzyme were unaffected by the
 exposures.  Leukocyte levels of alkaline phosphatase were unaffected at
 20 and 50 ppm,  showed  a dose-dependent increase at 100 and 300 ppm
 (P < 0.01), but  increased no  further at 1,000 or 3,000 ppm. The elevated
 enzyme levels were  observed following exposure to benzene,  but not to
 the other solvents.  As the  enzyme levels increased,  the leukocyte levels
 decreased.  Both  effects were  alleviated by administration of toluene.
 These results suggest  a relationship between benzene-induced increased
 enzyme levels and  leukopenia.

     Ward et  al.  (1985)  demonstrated changes in the peripheral blood and
 bone marrow of CD-I  mice  and  Sprague-Oawley rats exposed subchronically
 to 300 ppm  benzene.  Fifty male  and  50 female rats per group and 150 mice
 per group were exposed to 1,  10, 30, and 300 ppm benzene vapor, 6 h/day,
 5 days/week,  for  13  weeks and were  sacrificed at various time points up
 to 91 days  after exposure.  Hematological effects were not observed at 1,
 10, or 30 ppm. At  300  ppm,  however, male and female mice exhibited
 significant increases  (P  <  0.05) in mean cell volumes and mean cell
 hemoglobin  values and  decreases (P < 0.05) in hematocrit,  hemoglobin,
 RfiC count,  leukocyte count, platelet count,  and percentage of
 lymphocytes. These changes  were first observed on day 14 or day 28 and,
 in the males only, persisted  to the end of the study. The most common
 compound-related histological  findings included: myeloid hypoplasia of
 the bone marrow, splenic  periarteriolar lymphoid sheath depletion,
 lymphoid depletion in  the mesenteric lymph node, increased
 extramedullary hematopoiesis  in the spleen,  and plasma cell infiltration
 of the mandibular lymph node. All of these lesions were present at early
 sacrifice time points  and persisted throughout the study,  increasing in
 severity and  incidence  with time.  The effects were present more often in
males than  in females  and were more severe in males.

     In the rats, less  severely affected than the mice, the major
hematological effects,   statistically significant at 300 ppm (P < 0.05),
 consisted of decreased leukocyte counts in males on day 14 and in
 females on day 91 and  decreases in percentage of lymphocytes in both
males and females on days  14  through 91. The only exposure-related
histological lesion observed was a slightly decreased femoral marrow
cellularity.

-------
                                                 lexicological Daca   51

     Wolf et al. (1956) studied the effects of repeated inhalation
exposure to benzene on male rats. Benzene levels of 80 ppm (175
exposures for rabbits) or 88 ppm (136 exposures for rats.  193 exposures
for guinea pigs) induced leukopenia, increases in average spleen
weights, and histopathological changes in the bone marrow in the rats,
guinea pigs, and/or rabbits. Based on these results, the investigators
suggested that for humans the safe level for repeated vapor exposure to
benzene is well below 80 ppm. In addition to benzene, Wolf et al.  (1956)
tested eight alkyl benzenes, including toluene, styrene, and vinyl
toluene. Of the nine, benzene appeared to be the most toxic and was the
only chemical that affected the hematopoietic system.

     Other examples of characteristic hematological symptoms of benzene
poisoning and the exposures that produced them include: leukocytopenia
(100 ppm, continuous exposure for 2 days) (Gill et al. 1980);
lymphocytopenia (400 ppm. 6 h/day for 9.5 weeks) (Cronkite et al.  1982),
and in some cases granulocytopenia (4,000 ppm, 6 h/day produced "rapid"
decrease) (Gill et al. 1980) or transient granulocytosis (300 ppm,
6 h/day for 4 weeks) (C. A. Snyder et al. 1978).
     Benzene - induced changes in the peripheral blood reflect injury to
the cells of the hematopoietic organs. Special assays have been
developed to detect changes in leukocytes and erythrocytes at various
stages of development; these assays have been used extensively to study
the mechanistic aspects of benzene - induced hematotoxicity.

     Toft et al. (1982), using mice', demonstrated that benzene
concentrations in the range of occupational exposure levels reduced the
number of nucleated cells in marrow per  tibia and the number of colony-
forming granulopoietic stem cells (CFTI-C) per tibia. Continuous exposure
to 21 ppm benzene for 4 to 10 days significantly (P < 0.05) reduced boch
parameters, and intermittent exposure (8 h/day, 5 days/week for 2 weeks)
to 21 ppm significantly (P < 0.05) reduced the CFU-C per tibia. The
results were dose dependent. Toft et al. also observed that short and
high exposures had minimal effects on the parameters studied.
     Cronkite et al. (1985) demonstrated similar depression and,  in
addition, recovery of the pluripotential stem cells of the bone marrow.
C57B/6 BNL mice were exposed to 10, 25,  100,  300, or 400 ppm benzene for
2 to 16 weeks. Exposure to 10 and 25 ppm for  2 weeks did not result in
adverse effects, but exposure to 100 ppm significantly reduced overall
cellularity and the number of pluripotential  stem cells of  the bone
marrow  (P < 0.003 and P < 0.001, respectively). The peripheral blood
lymphocyte count was not affected at 10  ppm, but showed a dose-related
reduction aC the higher doses. Granulocyte levels were unaffected.  Stem
cell recovery was studied  in the 300-ppm exposure group.  In the animals
exposed  for 2 and 4 weeks  the stem cell  numbers had returned  to control
levels by 2 weeks after exposure, those  exposed for 8  weeks had
recovered by 16 weeks,  and  those exposed for  16 weeks  had recovered
incompletely to 92% of  control values by 25 weeks.  Lymphocyte  levels in
the peripheral blood of all  exposure groups  (300 ppm)  eventually
recovered to normal. The animals began  dying  about  9  months after
exposure, mainly of  thymic  and nonthymic lymphomas.

-------
 52   Section 4

      Other investigators have demonstrated similar pluripotential stem
 cell depletions in mice (Gill et al. 1980, Green et al.  1981b, C. A.
 Snyder et al. 1981b, Cronkite et al. 1982), as well as reductions in the
 granulocyte/macrophage progenitor cells (Green et al.  1981b,  C.  A.
 Snyder et al. 1981b).  The benzene metabolites benzene  dihydrodiol,
 hydroquinone, and phenol'did not affect bone marrow cellularity  or
 progenitor cell (CFU-C) numbers as severely as did benzene (Tunek et  al
 1981).

      In studies on the erythroid cell line,  Longacre et  al.  (1981b)
 demonstrated 60 to 80% decreases in the incorporation  of 59Fe into  the
 bone marrow precursors in mice injected subcutaneously with  6 to 20
 doses of 440 or 880 mg/kg benzene.  The effect was dose related.  Bolcsak
 and Nerland (1983) also demonstrated that  benzene and, to  a  lesser
 extent,  its metabolites phenol,  catechol,  and hydroquinone significantly
 decreased 59Fe incorporation into developing erythrocytes. Baarson et
 al.  (1984)  and Valle-Paul and Snyder (1986)  demonstrated that repeated
 exposures to 10 ppm benzene reduce  the progenitor red  cells  (CFTJ-E) in
 mice.

      Oral,  human.   No  data were  found for  benzene-induced  hematotoxicicy
 in humans exposed by the oral route.

      Oral,  animal.   Wolf et al.  (1956)  reported dose-related
 hematological effects  in female  rats administered repeated oral  doses of
 benzene.  Doses of 1,  10,  50,  and 100 mg/kg/day were  administered via
 oral intubation for -26 weeks. Olive-oil-treated rats  served  as  vehicle
 controls.  Leukopenia and erythrocytopenia  were observed  at the two
 highest  doses,  slight  leukopenia was observed at the 10-mg/kg dose, and
 there was no  effect at the  1-mg/kg  dose.

      Similar  results were  reported  in a more  recent  oral study (NTP
 1986). F344/N rats  and B6C3F1 mice  received doses  of 0,  25, 50,  100,
 200,  400, or  600  mg/kg in  corn oil  for  17  weeks  (NTP 1986). The  rats
 exhibited dose-related leukopenia,  lymphoid depletion  in the  spleen at
 200  mg/kg,  and increased extramedullary hematopoiesis  in the  spleen at
 600  mg/kg (120  days of exposure). Mice  in  the  400- and 600-mg/kg groups
 had  a dose-related  leukopenia.

     As  in  subchronic  studies, oral  administration of benzene  to
 C57BL/6N  mice  and F344 rats at doses  of 0, 50,  100, or 200 mg/kg,
 5 days/week,  for  103 weeks  resulted  in  dose-related lymphocytopenia and
 leukocytopenia  in both species (NTP  1986). The mice, in addition, had
 lymphoid  depletion  of  the splenic follicles and  thymus and hyperplasia
 of the bone marrow.

     Dermal, human.  No data  were found regarding  hematotoxicity  in
humans exposed dermally to benzene.

     Dermal, animal.  No data were found regarding hematotoxicity in
animals exposed dermally to benzene.

     General discussion.  Hematotoxicity is not a  significant concern  in
cases of acute exposure to benzene. However, in light of the serious
nononcogenic and oncogenic hematopoietic effects observed in humans
exposed subchronically and chronically  to  the chemical, numerous
experimental studies have focused on  the bone marrow.

-------
                                                 Toxicological Data   53

     Because of a paucity of data on the relationship between exposure
and effect in human studies and the variety of conditions under which
exposure occurs, it is difficult to determine from the literature the
lowest dose at which adverse effects can be observed in humans. However,
Chang (1972) studied 119 workers exposed to benzene concentrations of
-20 ppm and related'henratological changes to concentration and duration
of exposure. From these data he has suggested that benzene poisoning may
occur at levels as low as 10.1 ppm. American Conference of Governmental
Industrial Hygienists (ACGIH) estimates, however, are higher.

     The progression and consequences of benzene-induced pancytopenias
are very similar to those of idiopathic aplastic anemia (Goldstein
1977). Three possible exceptions to this are fairly high incidences of
bone marrow hyperplasia, lymphocytopenia, and macrocytosis that have
been observed in benzene anemia, but not in idiopathic anemia.

     The benzene-induced cytopenias, which can occur as a group or in
various combinations, may be related to specific adverse health effects.
For example, a decrease in circulating granulocytes diminishes the
bodily defenses against infections; this may account for the fact that
many victims of benzene toxicity have died from infections (Goldstein
1977). Thrombocytopenia, another cause of death in benzene poisoning,
induces capillary fragility, petechiae, subcutaneous bleeding
(bruising), or frank bleeding. Lymphocytopenia and eosinophilia, which
may be related to alterations in immune function, have been reported in
some cases.

     Evidence is accumulating for a positive association between
pancytopenia or aplastic anemia and later development of leukemia.
Vigliani and Saita (1964) described 13 cases of "benzene leukemia," of
which 9 were hemocytoblastic (or myeloblastic), 1 was chronic myeloid, 2
were erythremia, and 1 was erythroleukemia. In all cases in which a long
and severe exposure to benzene could be established, the leukemia was
hemocytoblastic in type, frequently preceded by aplastic anemia with
leukopenia. Aksoy and Erdem (1978) followed 44 benzene-exposed patients
with pancytopenia. The workers had been exposed to high concentrations
of benzene (ISO to 650 ppm) for 4 months to 15 years. Leukemia (the type
was not described) developed in six of them within 6 years of follow-up.
Other evidence of a relationship between pancytopenia and leukemia has
been reported (Hernberg et al. 1966, Vigliani and Forni 1976, and Aksoy
1981, all as reported in C. A. Snyder 1987; DeGowin 1963; Aksoy et al.
1976). According to Aksoy (1978), aplastic anemia is detected in
subjects generally while they are still exposed to high concentrations
of benzene; leukemia may occur at the same time or shortly after
exposure has ceased; however, in a few cases, the latency period between
exposure and the onset of leukemia has been long.

     The potential mechanisms for the development of pancytopenia in
humans include the destruction of bone marrow stem cells, the impairment
of the differentiation of these cells, or the destruction of more mature
hematopoietic cell precursors and circulating cells (Goldstein 1977).
Pancytopenia can also result from the combined destruction of the
peripheral blood and bone marrow elements. These mechanisms have been
explored in animal experiments.

-------
 54   Section 4

     The  role of  che microenvironment  In the depression of the colony-
 forming units of  the bone marrow has also been investigated. There is
 limited evidence  that  the stromal cells, particularly macrophages, may
 be  targets  for benzene - induced myelosuppression (Kalf et al. 1987). This
 was  suggested by  the results of an acute study by Frash et al. (1976),
 in which  the colony-forming potential  of bone marrow cells was not
 affected  when the cells were incubated with benzene prior to injection
 into a host animal, but was decreased  when benzene was injected into
 lethally  irradiated recipients after the normal bone marrow cells were
 injected. Gaido and Wierda (1984) demonstrated that benzene metabolites,
 particularly hydroquinone and benzoquinone,  were toxic to stromal cells
 in vitro  and that the  stromal toxicity resulted in impaired
 granulocyte/monocyte colony formation.

     Several molecular mechanisms for  myelotoxicity have been proposed.
 These include suppression of RNA and DNA synthesis (Post et al. 1985,
 Moeschlin and Speck 1967), alkylation  of cellular sulfhydryl groups
 (Irons 1985), disruption of the cell cycle (R.  Snyder et al. 1981, Irons
 et al. 1979), oxygen activation (or free radical formation) (Irons
 1985), and  covalent binding of benzene metabolites to cellular
 macromolecules (R. Snyder et al. 1978, Gill and Ahmed 1981, Longacre et
 al.  1981a).

     Previous sections in this report have demonstrated that benzene and
 its  metabolites localize in the bone marrow and that the metabolites are
 responsible for various hematotoxic effects.  When several of the
 metabolites of benzene were compared with the parent compound in
 hematotoxicity studies, the metabolites were less toxic than benzene.
 Tunek et  al. (1981) suggested that perhaps the metabolites tested may
 conjugate strongly in the liver and other organs and may not reach the
 bone marrow in amounts sufficient to produce the expected effect.  On the
 other hand, one metabolite of benzene, Crans.Crans-muconaldehyde (LD5Q,
 6.7  to 7.1 mg/kg). was highly toxic to mice when administered at the
 dose of 2 mg/kgi  intraperitoneally, for 10 to 16 days (Witz et al.
 1985).  The  treatment resulted in a statistically significant decrease in
RBC  count, hematocrit,  hemoglobin, bone marrow cellularity, and hepatic
 total and free sulfhydryl content. There was a dramatic increase in UBC
and  spleen weights at 16 days.

     It has, also been shown that agents that alter benzene metabolism
also modify benzene toxicity. Ethanol, which accelerates the
hydroxylation of benzene and transforms phenol into highly toxic
metabolites (Sato et al. 1980,  Nakajima et al.  1985), has increased the
severity of benzene-induced anemia, lymphocytopenia,  and reduction in
bone marrow cellularity and has produced transient increase in
normoblasts in the peripheral blood and atypical cellular morphology
 (C.  A.  Snyder et al.  1981a, Baarson et al.  1982, Nakajima et al.  1985).
The modulating effects of ethanol are dose dependent (Sato et al.  1981).
The enhancement of the hematotoxic effects of benzene by ethanol is of
particular concern for benzene-exposed workers who consume alcohol.

4.3.2.2  Imaunotoxicity

     Overview.   Lymphocytes play an important role in the immune
response.  Benzene - induced hematotoxicity involves the erythroid,

-------
                                                 lexicological Data   55

myelold, and lymphold lineages, of which Che lymphoid line appears co be
the most sensitive. Some of the manifestations of benzene-induced
immunotoxicity are reviewed in the following sections.

     Alterations in serum immunoglobulins and complement levels and
indications of benzene-induced autoImmunity and allergy have been
observed in benzene-exposed workers. Animal studies support the findings
of immune dysfunction in humans and further define the various
parameters of the immune response that are altered in response to
benzene exposure. These studies show that the components responsible for
antibody production (B-cells) and those responsible for cell-mediated
immunity (T-cells) are significantly depressed by benzene concentrations
as low as 10 ppm, depending on the parameter measured. Impaired cell-
mediated immunity results in decreased resistance to infectious agents
and to transplanted tumor cells.

     Inhalation, human.  Altered serum immunoglobulins and complement
levels have been found in workers exposed to benzene and other solvents
(Lange et al.  1973a,b; Roth et al. 1972; Smolik et al. 1973, all as
reported in Goldstein 1977). Renova (1962, as reported in Goldstein
1977) detected antibodies against leukocytes, platelets, and red cells
in the sera of chronically exposed workers.

     Symptoms of immune stimulation (allergy) have been reported in
workers chronically exposed to benzene. For example, eosinophilia, an
indication of an allergic response, has been noted by several
investigators (Aksoy et al. 1971, Hernberg et al. 1966, Bernard and
Basset 1946, Blaney 1950, Duvoir and Derobert 1942, all as reported in
Goldstein 1977). Lange et al. (1973a, as reported in Goldstein 1977)
reported a positive leukocyte autoagglutinin test associated with
decreased granulocyte levels, suggestive of allergic blood dyscrasia.
Roth et al. (1973, as reported in Goldstein 1977) emphasized the role of
reticulosis and autoimmune phenomena in the pathogenesis of bone marrow
damage.
     Inhalation, animal.  Rozen et al. (1984) demonstrated in a dose-
response study that short-term inhalation exposure (6 h/day for 6 days)
to benzene at near occupational exposure levels significantly depressed
the proliferative responses of bone-marrow-derived B-cells and splenic
T-cells. CS7B1 mice were exposed to benzene concentrations of 0, 10, 30.
100, or 300 ppm. The lipopolysaccharide-induced B-lymphocyte
proliferative response was depressed (P £ O.OS) at 10 ppm, and the
phytohemagglutinin (PHA)-induced T-cell response was depressed
(P £ O.OS) at 31 ppm, without causing a significant concomitant
depression in the numbers of T- or B-cells. Peripheral lymphocyte counts
were depressed at all levels, but erythrocyte counts were depressed only
at 100 and 300 ppm.

     Following the exposure regimen that previously had induced thymic
lymphoma in mice, Rozen and Snyder (198S) then demonstrated that benzene
concentrations of 300 ppm 6 h/day for 115 exposures reduced the
abilities of T- and B-cells to respond to mitogenic stimuli and markedly
reduced the numbers of B-lymphocytes in the bone marrow and spleen and
the number of T-lymphocytes in the thymus and spleen  (Rozen and Snyder
1985). In addition, a compensatory proliferation was  observed  in cells
of the bone marrow and thymus  in response to the benzene exposures.

-------
 56   Section 4

      Benzene suppressed the primary antibody response to fluid tetanus
 toxoid (FTT) by 74 to 89% in Swiss albino mice exposed to 400 ppm of the
 chemical 6 h/day for 5, 12, or 22 exposures (Stoner et al.  1981).
 Similar exposure to adsorbed tetanus toxoid (APTT)  for 5, 12, or 22 days
 also suppressed the primary response,  by 8, 36.  and 85%,  respectively.
 Significant suppression of the responses to both antigens was also seen
 with exposures to 200 ppm benzene for  10 to 20 days,  but no significant
 effect was seen at 50 ppm. At benzene  concentrations  of 50,  200,  or
 400 ppm,  the secondary antibody response to tetanus antitoxin was
 unaffected. The investigators concluded that the threshold  dose  for
 repression of the primary antitoxin responses is between 50 and  200 ppm
 inhaled benzene.

      Rosenthal and Snyder (1985)  studied the effect of subacute  exposure
 to benzene on cell-mediated immunity by measuring host resistance to
 infectious agents.  Mice were exposed to benzene  concentrations of 10,
 30,  100,  and 300  ppm for either 5 days  prior to  (preexposure regimen)  or
 5  days prior to and 7 days during (continuous exposure regimen)
 infection with Lisceria monocycogenes.  On days 1, 4,  and  7  of infection,
 splenic bacterial counts were made,  and T-  and B-lymphocytes were
 enumerated by a direct immunofluorescence technique.  These  results were
 compared  with those of appropriate air  controls.

      Preexposure  to the benzene concentration of 300  ppm  resulted in
 increased bacterial numbers  (730% of controls) on day 4.  Lower
 concentrations  of benzene  had no  such effect.  Continuous  exposure,  on
 the  other hand, produced increased bacterial  counts in the  spleen on
 day  4  at  all but  the  10-ppm  concentration.  Concentrations of 30,  100,
 and  300 ppm increased the  bacterial  counts  to  490,  750, and  720%  of
 controls,  respectively.  Bacterial counts  were  not increased  on days 1
 and  7  at  any benzene  concentration for  both exposure  regimens,
 indicating recovery of the immune response  at  day 7.  This indicates a
 delay  in  the cell-mediated immune response.  In addition, a
 concentration-dependent depression was  noted  in  T- and B-lymphocyte
 populations  (P £  0.05  at £30  ppm). B-cells  were  more  sensitive to
 benzene than were T-cells  on  a  percentage of control  basis.

     Rosenthal and  Snyder  (1986)  tested another  parameter of cell-
 mediated  immunity,  that of tumor  resistance. Male C57B6J mice were
 exposed to  100 ppm benzene (6 h/day, 5  days/week, for 20 exposures)  and
 were then  injected with cells from a virus-induced tumor. Ninety  percent
 of the benzene-treated mice developed lethal tumors,  in comparison  with
 only 30% of  the non-benzene-treated controls.

     Other routes of exposure.  No data were found to  document
 immunetoxicity in humans or animals exposed orally,  or by dermal
 application,  to benzene.

     The various parameters and mechanisms of benzene  immunotoxicity
have been investigated  in  animals  injected with  the chemical
 subcutaneously and intraperitoneally, as well as in in vitro
experiments. These Important studies support the results of  inhalation
 studies in demonstrating that (1) benzene depresses antibody formation
 in mouse spleen cells, reflecting alterations in spleen cell function
 (Vierda et al. 1981);  (2)  benzene produces a selective reduction  in the
number of B-lymphocytes  in the circulating blood of rabbits  (Irons  and

-------
                                                 lexicological Data   57 -

Moore 1980); and  (3) metabolites of benzene  (catechol, hydroquinone,
1.2,4-benzotriol, and benzoquinone) are cytotoxic to spleen cells,
reduce the number of progenitor cells from the spleen and bone marrow,
and/or suppress T- and B-lymphocyte mitogen  responses (Wierda and Irons
1981, Wierda et al. 1980, both as reported in Vierda et al. 1981; Irons
et al. 1981; Pfeifer and Irons 1981).

     General discussion.  The immune system, a complex and widespread
organization of organs and cells, is responsible for allergic reactions,
tumor immunity, host-graft reactions, immunological tolerance, the
defense of the body against microbial infections, and the pathogenesis
of various occupational disorders of the lung and skin.  Injury to the
immune system can have serious health consequences.

     An important function of the immune system is the immunosurveillance
of carcinogenesis, in which lymphocytes are  thought to play an important
role. Goldstein (1977) and Leong (1977) postulated that benzene
leukemogenesis could be a result of the impairment of this mechanism.

     Rozen and Snyder (1985) observed a compensatory proliferative
response of the cells of the bone marrow and thymus of animals following
6 months exposure to benzene at a concentration that had previously
produced thymic lymphoma in mice. The investigators suggested that the
response may play a role in benzene carcinogenicity.

     Suppression of cell growth and function in the lymphoid system,  as
in the bone marrow, correlates with the concentrations of hydroquinone
and catechol which accumulate in lymphoid tissue following exposure to
benzene (Greenlee and Irons 1981, Greenlee et al. 1981,  Irons et al.
1980b, Wierda and Irons 1982). Hydroquinone, p-benzoquinone, phenol,  and
catechol also suppress microtubule assembly and progenitor cells (Kalf
et al. 1987).

     The suppression of phytohemagglutinin-stimulated lymphocyte
activation by the metabolites of benzene may be mediated through the
inhibition of microtubule function; the inactivation correlates with  the
ability of the metabolites to undergo sulfhydryl-dependent autooxidation
(Irons et al. 1981, Pfeifer and Irons 1981). Irons and Neptun (1980)  and
Irons et al. (1981) suggested that hydroquinone or its terminal
oxidation product, p-benzoquinone, may be responsible for these effects.

4.3.2.3  Neurotoxicity

     Inhalation, human.  Following acute inhalation of benzene,
individuals exhibit symptoms indicative of CNS toxicity (Sandmeyer
1981). These include drowsiness, dizziness, headache, vertigo, and
delirium and perhaps loss of consciousness.  Symptoms are similar in
lethal and nonlethal exposures (EPA 1980b). The neurological effects  of
benzene are thought to be direct effects of benzene rather than  its
metabolites (Bergman 1979, as reported in Dempster et al. 1984).

     At low chronic exposure levels, workers have experienced symptoms
of CNS lesions [i.e, dizziness when cold water is placed in the  ear and
impairment of hearing (Brzecki et al. 1973,  as reported in Sandmeyer
1981)]. Workers exposed to benzene in combination with other chemicals
exhibited asthenoneurotic or astheno-vegetative polyneuritis, sometimes
associated with neuronal progression, even after exposure had ceased

-------
 58    Section 4

 (Drogichina et al.  1971,  as reported in  Sandmeyer  1981).  it  is  possible,
 however,  that CNS involvement in benzene toxicity  is generally  masked by
 other,  more visible,  effects (Sandmeyer  1981).

      Inhalation,  animal.   Disturbed neuronal transport characteristics
 have  been noted in animals following acute inhalation of  benzene  (Jonek
 et  al.  1965,  as reported  in Sandmeyer 1981).

      Rabbits  exposed  to -45,000  ppm benzene exhibited light narcosis
 (relaxation)  after 3.7 min of exposure;  tremors, chewing, excitement,
 and running movements after 5 min;  loss  of pupillary reflex to  strong
 light after 6.5 min;  loss of blink  reflex to tactual stimulus after
 11.4  min;  pupillary contraction  after 12 min; involuntary blinking after
 15.6  min;  and death after 36.2 min  (Carpenter et al. 1944).

      Dempster et  al.  (1984)  demonstrated behavioral disturbances,
 characterized by  increased milk-licking, in mice exposed  to benzene
 concentrations of 100 ppm after  1 day of exposure and to  300 ppm after
 5 days  of exposure. Less  sensitive  parameters,  home-cage  food intake and
 hind-limb grip strength,  were reduced only at 1,000 and 3,000 ppm. The
 mice  were exposed to  concentrations  of 100, 300, 1,000,  and 3,000 ppm
 6 h/day for the number of days necessary to reach a minimum
 concentration x time  product of  3,000  ppm-days.  The milk-licking change
 occurred  at the same  time as hematological changes, suggesting  that the
 behavioral  change may be  mediated by the metabolites of benzene, as are
 hematological  changes.

      Other  routes of  exposure.  Geist  et al.  (1983) demonstrated
 learning  deficits  in  Sprague-Dawley  rats given oral doses of 550 mgAg
 benzene.  No data were found  demonstrating neurological effects  in humans
 or  animals  exposed  to benzene  via the  dermal route. However,  CNS effects
 have been reported  in animals  following intravenous injection (Braier
 and Francone  1950,  as reported in Sandmeyer 1981).

     General discussion.   The  neurotoxicity of benzene has not been
 studied extensively. The  most  obvious  effects that occur following acute
 exposure  to very high concentrations are fairly well documented, but the
 more subtle effects that  might occur  from chronic exposure to low
 concentrations  may be masked by or overlooked because of the alterations
 that  take place in  the hematopoietic system.

 4.3.2.4  Dermal toxicity

     Wolf et al.  (1956) reported that benzene was slightly to moderately
 irritating  to  the skin of rabbits and caused moderate necrosis.  Ten to
 20  applications of  the undiluted chemical to the ear (method of Adams et
 al.  1941, as reported in  Wolf  et al. 1956) produced erythema, edema,
 exfoliation, and blistering.

 4.3.2.5  Ocular toxicity

     Wolf et al.  (1956) tested benzene for irritation in the rabbit eye.
Two drops of the undiluted material  caused moderate conjunctival
 Irritation and very slight,  transient corneal injury. There were very
 small areas of  superficial necrosis  in the cornea.

-------
                                                 lexicological Data   59

     Information was not found for benzene toxicity in other major organ
systems.

4.3.3  Developmental Toxicity

4.3.3.1  Overview
     Although there is little information on the developmental toxicity
of benzene in humans, benzene has been shown to be embryo/fetotoxic in
animals, as evidenced by increased incidences of resorptions,  reduced
fetal weight, skeletal variations, and altered fetal hematopoiesis.
However, benzene has not been shown to be teratogenic or embryolethal in
test animals even at levels causing toxicity to the mother,  as evidenced
by reduced weight gain. Humans are most often exposed to benzene by
inhalation. This route of exposure has therefore been used in
developmental animal research more than other routes, with the rat as
the most common test animal.

4.3.3.2  Inhalation
     Human.  Benzene crosses the human placenta and is present in the
cord blood in amounts equal to those in maternal blood (Dowty et al.
1976). In a study of subjects with known benzene poisoning in Italy,
Forni et al. (1971b) report the case of one pregnant worker exposed to
benzene during her entire pregnancy. Although she had severe
pancytopenia and increased chromosomal aberrations, she delivered a
healthy boy with no evidence of chromosomal alterations. The following
year she delivered a normal daughter. Sometimes a normal infant survives
when the mother dies at parturition of severe anemia caused by benzene
(Messerschmitt 1972) . An increased frequency of chromatid and
isochromatid breaks and SCE was found in lymphocytes from 14 children of
female workers exposed to benzene and other organic solvents during
pregnancy (Funes-Cravioto et al. 1977). No mention was made of physical
abnormalities among the offspring. Holmberg (1979) reported the
occurrence of congenital CNS defects in children of 14 mothers exposed
to organic solvents during the first trimester of pregnancy. Only one of
these women was exposed to benzene, and she gave birth to a stillborn
anencephalic fetus; however, she was also exposed to other solvents.
Epidemiological studies of pregnant women occupationally exposed to
undefined organic solvents found no effect on birth weight (Olsen
1983a), on frequency of malformations (Olsen 1983b), or on rates of
perinatal death or malformations  (Axelsson et al. 1984). In a study
conducted in the Love Canal area by Heath (1983), the outcome of
pregnancy was evaluated in populations living in the proximity of waste
dumps in which benzene had been identified. No clear increased
occurrence of abortion, birth defects, or low infant birth weight was
observed in women living next to  the canal. In a study by Budnick et al
(1984), no statistically significant clusters of birth defects were
found when analyzing data from Drake Superfund Site, Pennsylvania, an
area contaminated with benzene and other carcinogens.

     Animal.  The many studies of exposure by inhalation during
pregnancy have shown that benzene is not teratogenic, even when  the
mother and offspring show signs of  toxicity. Fetotoxicity is  evidenced
by decreased weight and by  an increase in skeletal  variants such as

-------
 60   Section 4

 missing sternebrae and extra ribs, which are not considered to be
 malformations (Kimmel and Wilson 1973). Mouse and rabbit studies are
 summarized in Table 4.1; rat studies are summarized in Table 4.2.

      Murray ec al. (1979) exposed mice to 500 ppm benzene for 7 h/day on
 days 6 to 15 of pregnancy and observed growth retardation and increased
 skeletal variants but no malformations in fetuses and no significant
 effect on incidence of pregnancy,  average number of live fetuses, or
 resorptions per litter.  Similar results were found in rabbits exposed to
 the same level.  Ungvary and Tatrai (1985) exposed pregnant mice to 500
 or 1,000 mg/m3 (157 or 313 ppm) benzene on days  7 to 20 of gestation and
 noted a significant fetal weight retardation but no malformations.

      Alterations in hematopoiesis  have been observed in the fetuses and
 offspring-of pregnant mice exposed to benzene (Keller and Snyder 1986).
 Administration of 5,  10,  or 20 ppm benzene by inhalation to pregnant
 Swiss Webster mice for 6 h/day on  days 6 to 15 of gestation caused
 marked changes in the numbers of erythroid colony-forming cells of the
 progeny.  Granulocytic colony-forming cells were  affected by the two
 higher exposure  concentrations.  Some of the mice exposed prenatally were
 allowed to macure and were reexposed.  Their hematopoietic progenitor
 cell  numbers  were depressed compared with controls  exposed for  the  first
 time  as adults.  Other hematopoietic end points were affected in a
 previous  study (Keller et al.  1985),  where pregnant mice were exposed to
 only  one  level of benzene,  10 ppm.

      The  many studies using rats provide ample data for  comparison  of
 results,  but  the most helpful ones  are by Kuna and  Kapp  (1981),  Coate et
 al. (1984), Green et  al.  (1978), and Tatrai et al.  (1980a).  Table 4.2
 summarizes results when  exposure occurred essentially only during
 organogenesis. The data  are arranged by level of exposure  and show  a
 dose  response. In the following paragraphs  the various studies  will be
 discussed by  end point.

      Maternal toxicity, as  indicated by a decrease  in maternal  weight
 gain,  is  clearly evident  at levels  of more  than  100  ppm. Resorption data
 from  the  eight rat studies  are  inconclusive, because  of  the  divergent
 results.  Green et al.  (1978)  found  no  increase in resorptions at  the
 2,200-ppm exposure level. The studies  at  Litton  Bionetics  Incorporated
 (Litton 1977  and 1978, as reported  by  Schwetz 1983)  showed an increase
 in resorpcions at relatively  low levels  of benzene exposure, but  Schwetz
 observed  that  in the  1977 study  an  air  temperature rise  in one  of the
 chambers might have produced maternal  stress that increased  resorptions.
 In the  1978 study,  the experimental  control value of  2%  resorptions was
 unusually  low, skewing statistical  results. The  rate of  resorptions  at
 10 and  40  ppm was  comparable  to historical controls.

      The  fetotoxicity of benzene in  rats  is demonstrated by  retarded
 fetal weight and  skeletal variants. Table 4.2 shows  that exposure to
high  levels of benzene ranging from  50  to 2,200 ppm resulted in a
 significant decrease  In fetal weight, with the exception of  two groups
of rats in a study by Green et al.   (1978). In this study, dams breathing
air containing 100 or 300 ppm benzene bore young which were similar  in
weight  to control pups. Statistically significant numbers of skeletal
variations (delayed ossification, missing sternebrae, and extra ribs)

-------
                                                           Toxicologies!  Data     61
Table 4.1. Summary of results of some teratological studies on benzene in (be moose and rabbit0
Animals Route
New Zealand Inhalation
rabbit
New Zealand Inhalation
rabbit
CF-1 mouse Inhalation
CFLP mouse Inhalation
ICR mouse Subcutaneous
CD-I mouse Gavage
Exposure
level
500 ppm
156 ppm
313 ppm
500 ppm
156 ppm
313 ppm
2 mL/kg
4 mL/kg
0 3 mL/kg
0 5 mL/kg
1 0 mL/kg
" — = No significant difference compared
pared with controls
* Resolutions
c Fetal death
''Abortions
Maternal Fetal
weight body
gain weight
-
DECR DECR
DECR
DECR
DECR
DECR.
DECR
DECR
DECR
with controls. DECR —
Resorptions/
fetal death/
abortions
-
INCR*
—
INCR*'C
INCR* c
INCR*'C
INCR*'C
INCR*
INCR*
Skeletal
variants Malformations
Slightly
INCR
INCR
INCR
INCR
INCR
INCR
— -
decreased compared with controls, INCR ~
References
Murray et al
1979
Ungvary and
Tatrai 1985
Murry et al
1979
Ungvary and
Tatrai 1985
Matsumoto et
al 1975
Nawrot and
Staples 1979
increased com-

-------
   62
Section 4
                                 Table 4.2,  Teratology studies on inhaled benzene in rats"
Strain
S-D
S-D
S-D
S-D
S-D
S-D
CFY
S-D
S-D
S-D
CFY
CFY
CFY
S-D
CFY
CFY
S-D
CFY
S-D
a _
Hours
per day
6
7
6
6
6
6
24
7
6
6

24
24
6
24
24
7
24
6
— No signi
Benzene
(ppm)
I
10
10
10
40
40
47
30
100
100
125
125
141
300
313
470
500
940
2200
ficant diffei
Maternal
weight gain
-
:
-
-
~
DECR
DECK
-
-
DECR
DECR
DECR
-
DECR
DECR
DECR
DECR
DECR
"ence consD&rei
Resorpttons
-
[NCR
-
INCR
-
-
-
-
-
-
-
INCR
-
-
INCR
-
INCR
—
! with controls:
Fetal
weight
-
^
-
-
-
DECR
DECR
-
DECR
DECR
DECR
DECR
-
DECR
DECR
DECR
DECR
DECR
DECR -
Skeletal
variants
-
	
-
-
-
-
INCR
-
-
INCR
INCR
INCR
INCR
INCR
INCR
INCR
INCR
INCR
decreased con
Malformation] References
- Coate et al 1984
Kuna and Kapp 1981
Litton6
Coate et al. 1984
— Litton*
Coate et al. 1984
Tatrai et al. 1980a
Kuna and Kapp 1981
— Green et al 1978
- Coate et al. 1984
Ungvary 1985
- Tatrai et al 19806
- Tatrai et al. 1980s
- Green et al. 1978
Hudak and Ungvary 1978
- Tatrai et al. I980a
Kuna and Kapp 1981
- Tatrai et al 1980a
- Green et al. 1978
wared with contrail: INCR — incrauai
compared with controls.
    Luton Bionetics Incorporated (Litton 1977 and 1978, as reported in Schweu 1983).

-------
                                                 lexicological Data   63

were found  in all groups exposed  to  concentration levels equal to
125 ppm and higher.

     Similar to the studies  cited in Table 4.1, those reported in
Table 4.2 failed to find malformations  as a result of benzene exposure
The highest level of exposure was 2,200 ppm (Green et al. 1978). One
exencephalic rat was found in a group exposed to 500 ppm benzene at the
Hazelton Laboratories  (Kuna  and Kapp 1981), where 151 pups were
examined. In the same  study, out  of  98  pups examined for skeletal
defects, one pup had angulated ribs  and two other pups had nonsequential
ossification of the forefeet. These  anomalies were not statistically
significant and may have resulted from  maternal nutritional stress.
Information from the control group was  inadequate, because only 11 of
the 17 control dams produced offspring. Since other studies using high
levels of benzene did  not result  in  these malformations, there does noc
appear to be significant evidence that  they were caused by benzene.

     In two studies not included  in  Table 4.2, rats were exposed to
inhaled benzene both pre- and postconceptionally (Table 4.2 was
restricted  to postconceptional exposure only). Gofmekler (1968) showed
that continuous exposure of  female rats to 0.3 to 20 ppm benzene for 10
to 15 days before mating and throughout pregnancy did not affect newborn
weight, but there were differences in the weights of individual organs.
There was a slight tendency  toward decreased litter sizes at higher
levels of exposure. A  complete absence  of litters resulted from exposure
to 210 ppm for 10 to 15 days before  mating and throughout pregnancy.
Pushkina et al. (1968) exposed female rats for 20 days before mating and
throughout pregnancy.  No malformations  resulted. Six different
concentrations of benzene ranged  from 0.3 to 210 ppm, and at the higher
levels the litter size was smaller.  The weight of the fetuses decreased,
and the relative weights of their organs were reduced.

4.3.3.3  Oral

     Human.   No information was found.

     Animal.  Benzene  administered orally to CD-I mice at 0.3, 0.5, or
1.0 mLAg on days 6 to 15 of pregnancy  caused maternal lethality and
resorptions at the 0.5- and 1.0-mLAg levels but no increase in
malformations (Nawrot  and Staples 1979). Seidenberg et al. (1986) gave
pregnant mice 1300 mgAg/day (1-47 mLAg/day) benzene by gavage on
gestation days 8 to 12. A significant reduction in fetal body weight was
observed.

4.3.3.4  Dermal

     Human.   No information was found.

     Animal.  Prolonged application  of  benzene to the skin of rats for
4 months at 64 or 320  mgAg/day increased the mortality of the first-
generation offspring (Malysheva 1980).

4.3.3.5  Injection

     Human.   No information was found.

-------
 64   Section 4

      Animal.  Guinea pigs treated with subcutaneous  Injections  of
 0.1  gAg/day (0.11 mL/kg/day)  of a 40% solution of benzene
 (0.044 mL/kg/day benzene) throughout pregnancy did not  show  any effect
 on maternal body weight,  length of gestation, number of offspring,
 offspring body weight,  or neonatal mortality  (Desollle  et al. 1967, as
 reported by Barlow and Sullivan 1982).  The same Investigators,  using
 rats,  injected 0.1 gAg/day (0.11 mLAg/day) benzene in a solution of
 olive  oil throughout pregnancy and found no effect on duration  of
 pregnancy,  maternal body  weight,  average litter size, or neonatal
 mortality rates (Desoille et al.  1965,  as reported by Barlow and
 Sullivan 1982).  No mention of  malformations was made in the guinea pig
 and  rat studies.  In a study by Matsumoto et al. (1975), ICR mice were
 administered benzene by the subcutaneous route at doses of 2 or  4 mL/kg
 on days 8 and 9 or 12 and 13 of gestation. An increased incidence of
 delayed ossification of bones  was found at the high dose level when
 given  on gestation days 12 and 13.  A decrease in white  cell counts and
 hemoglobin content in the mothers was consistent with expected effects
 of benzene  and may have contributed to  the decreased weight of  the
 fetuses.

 4.3.3.6  General  discussion

     Epidemiological studies implicating benzene as a developmental
 toxicant  in humans have many limitations. These include exposure to
 multiple  substances,  lack of control  incidences for end points,  problems
 in identifying exposed  populations,  and lack of data on exposure levels.
 Thus it is  extremely difficult to make  a clear assessment of the effect
 of benzene  on the human conceptus.  The  preponderance of animal data is
 from inhalation experiments, because  this has been the principal route
 of concern.  More  data are needed  on other routes,  since exposure from
 waste  sites  will  likely also involve  other routes. Inhalation results
 have been fairly  consistent across  species.  It has been suggested that
 benzene fetotoxicity in the rat appears to be a function of maternal
 toxicity, as the  Joint  occurrence  of  a  decrease in fetal weight and an
 increase  in  skeletal variants  usually occurs when there is a decrease in
 maternal  weight.  There  is no clear-cut  evidence on mechanisms of this
 toxicity, and there  are little data on maternal food consumption during
 benzene exposure  and blood levels  of  benzene and its metabolites in the
 mothers and  their fetuses.  There  is sufficient evidence that benzene is
 not teratogenic and  not overtly embryotoxic at 10 ppm.

     Schwetz (1983)  has provided  an excellent review of research on
 developmental  toxicity  in animals, while Barlow and Sullivan (1982) have
 provided  Che most comprehensive information on humans.

 4.3.4   Reproductive  Toxicity

 4.3.4.1   Overview

     There is  a paucity of information on the reproductive toxicity of
benzene.  Some  harmful gonadal  effects have been noted in experimental
 animals, but the  levels of exposure have been higher than those to which
humans  are exposed in the  modern  industrial environment.

-------
                                                  Toxicologlcal  Data    65

 4.3.4.2   Inhalation

      Human.   Vara and Klnnunen (1946)  examined 30 women with  symptoms  of
 benzene  toxlcity Indicating exposure to levels much higher  than those  In
 today's  working environment.  Twelve  of these  women had  menstrual
 disorders  (profuse or scanty periods and dysmenorrhea).  Ten of  these 12
 women were married,  and two abortions  and no  births took place  during
 their employment even though no  contraceptive measures  had  been taken,
 leading  the  Investigators  to suggest a detrimental effect on  fertility
 at high  levels  of exposure.  Gynecological examinations  revealed that the
 scanty menstruations of five of  the  patients  were due to hypoplasia of
 the ovaries.  Menstrual disturbances  (heavy bleeding) were reported In
 female workers  exposed to  31  ppm benzene In a Polish factory  (Michon
 1965, as cited  in Barlow and Sullivan  1982).  In another group of factory
 workers, occupational benzene poisoning resulted  in ovarian hypofunction
 (Pushklna et  al.  1968).  In a  study of  360 female  gluing operators
 exposed  to petroleum (a major source of benzene)  and chlorinated
 hydrocarbons  both dermally and by inhalation,  no  significant difference
 in fertility  between exposed  workers and unexposed controls was  found
 (Mukhametova  and Vozovaya  1972,  as reported In Barlow and Sullivan
 1982). However,  spontaneous  abortion and premature birth increased.

     Animal.  In a subchronic inhalation study, Ward et  al. (1985)
 exposed male  or female CD-I mice to  concentrations of 1, 10, 30, or
 300 ppm benzene vapor,  6 h/day,  5 days/week for 13 weeks. Hlstopatholog-
 ical changes  were  observed In ovaries  (bilateral  cysts)  and testes
 (atrophy/degeneration,  decrease  in spermatozoa, moderate increase in
 abnormal sperm  forms)  of mice exposed  to 300  ppm  benzene; the severity
 of the lesions  was greater in the males.  An inhalation  study was con-
 ducted by Wolf  et  al.  (1956)  exposing  rabbits and guinea pigs to benzene
 7 to 8 h/day, 5 days/week,  for up to 6  months.  The guinea pigs showed  a
 slight increase in average testicular weight  at the 88-ppm  level.
 Rabbits showed  slight histopathological testicular changes  (degeneration
 of the germinal epithelium) when exposed to 80  ppm. Gofmekler (1968)
 found that continuous exposure of female rats to  210 ppm benzene for 10
 to 15 days before  mating and  3 weeks after mating resulted  in a complete
 absence of litters.  It is  not known  whether this  was due to failure to
mate,  infertility, or early preimplantation losses of fertilized ova.

4.3.4.3  Oral

     Human.   No studies are available.

     Animal.  No studies are  available.

4.3.4.4  Dermal

     Human.   No reproductive  data with  dermal as  the only route of
exposure to benzene  are available. Multiroute exposure was discussed in
 the section on  inhalation.

     Animal.  Prolonged dermal application of benzene to the tails of
adult rats for  4 months at 64 or 320 mg/kg/day  did not affect the
 fertilizing ability  of males  or  the  conceptional  capacity of females
when mated with untreated  rats (Malysheva 1980).  There was, however, a
decrease in the number of  spermatogonia in the  males.

-------
66   Section 4

4.3.4.5  Injection
     Human.  No studies are available.

     Animal.  An increased incidence of abnormal  sperm heads was induced
in male mice given five daily intraperitoneal  injections of benzene in
corn oil at doses ranging from 0.5 to 1.0 mLAs (Topham 1980).

4.3.4.6  General discussion
     As with studies of human developmental toxicity,  evidence  of an
effect of benzene exposure on human reproduction  is not sufficient to
demonstrate a definite association. Exposure to benzene occurs  along
with exposure to many other chemicals,  so no conclusion can be  drawn
relative to any single agent. There are insufficient animal data to
propose NOAELs and LOAELs.

4.3.5  Genotoxic ity

4.3.5.1  Overview
     Benzene (or more likely its metabolites)  causes both structural and
numerical chromosome aberrations in humans, laboratory animals, and
cells in culture and SCEs and micronuclei in in vivo animal studies.
Other positive effects found following in vivo animal exposure  include
sperm-head abnormalities, inhibition of DNA and RNA synthesis,  DNA
binding, and interference with cell cycle progression. DNA binding and
inhibition of DNA synthesis have also been demonstrated in vitro.
Benzene has rarely been shown to cause gene mutations.
     This review is not an exhaustive critique of all available
literature on the genetic toxicity of benzene but intends to present
sufficient data to support the conclusion that benzene is a genetic
hazard to humans. This section is organized by end point under  human,
animal, and in vitro headings rather than by route of administration.
This approach was taken for the following reasons: much of the
supporting data for human effects come from in vitro studies for which
route of exposure is not applicable; no in vivo animal studies
considered in this review used dermal exposure, while many used either
intraperitoneal or subcutaneous injections, routes not likely to be
important for human exposure but useful in determining potential
genotoxic effects; and virtually all human epidemiology studies report
only inhalational exposures, although concurrent dermal exposure cannot
be excluded.

4.3.5.2  Human

     Available evidence for benzene- induced genetic toxicity in humans
comes from epidemiological studies of persons occupationally exposed to
benzene. Exposures reported  in these studies are primarily via
inhalation, although contact with  the skin may also have occurred  in
some cases. The difficulties with  epidemiological studies are well known
and include lack of accurate exposure data, possible exposure  to
multiple chemicals, and selection  of appropriate control groups.
However, the association between benzene exposure and  the appearance of
structural and numerical chromosomal aberrations  in human lymphocytes

-------
                                                 Toxicological Data   67

has been found so consistently that benzene can undoubtedly be
considered a human clastogen. The evidence for benzene-induced
chromosomal aberrations 'in humans has been extensively reviewed (R.
Snyder et ml. 1977. Dean 1978, Forni 1979, White et ml.  1980, IARC 1982,
C. A. Snyder 1987, van Raalte and Grasso 1982, Dean 1985).
     Tough and Court Brown (1965) examined blood samples from 20 men
with benzene exposures of 1 to 20 years. Benzene exposure had stopped
2 years before, at a time when 14 of the men had shown neutropenia. The
number of lymphocytes with unstable chromosomal aberrations (Cu cells)
was significantly higher in the exposed men than in on-site or off-sice
controls (1.4% for exposed vs 0.6% for both sets of controls), while the
number of cells with stable aberrations (Cs cells) was higher than in
controls but not significantly higher. Pollini et al.  (1969) found 70%
aneuploid lymphocytes in five women with benzene hemopathy. This value
dropped to 40% 5 years later, when all five women still had stable and
unstable chromosome changes. Large metacentric chromosomes of group 1-3
were often missing. Forni et al. (1971a) performed chromosome studies on
lymphocytes of workers in a rotogravure plant where benzene was used as
a solvent. The workers were exposed to benzene for 1 to 22 years (until
an epidemic of benzene poisoning led to its replacement by toluene) and
then to toluene for an additional 12 to 14 years. At the time of benzene
poisoning, concentrations of 125 to 532 ppm benzene were measured in
various parts of the plant. Statistically significant increases in both
unstable and stable chromosomal aberrations were found in benzene -
exposed workers as compared with either unexposed controls or with
toluene-only-exposed controls. Another study by Forni et al. (1971b)
found similar results in a group of 25 persons (13 men and 12 women)
following recovery from benzene hemopathy. Exposure levels were unknown
but were obviously high enough to cause clinical symptoms, in some cases
quite severe.  Blood samples were taken 1 to 18 years following
recovery. The percentage of both Cs cells and Cu cells was significantly
higher in the exposed group than in the control group. Only 1 of 25
control individuals had >1% Cu cells and 1 had 1% Cs cells, compared
with 18 of 25 exposed individuals with >1% Cu cells and 17 of 25 with 1%
or more Cs cells. There was a tendency for the rate of unstable
chromosome aberrations to decrease and stable aberrations to increase or
remain steady in follow-up studies; however, there was no consistent
pattern of change. Other studies have consistently found increased
chromosomal aberrations in benzene-exposed populations (Erdogan and
Aksoy 1973, Khan and Khan 1973, Ding et al. 1983. Koizumi et al. 1979,
Van den Berghe et al. 1979). Additional case studies also support  the
chromosomal-damaging effect of benzene  (Hartwich et al. 1969, Forni and
Moreo 1967, Forni and Moreo 1969, Sellyei and Kelemen 1971).

4.3.5.3  Animal

     Benzene-induced cytogenetic effects, including chromosome and
chromatid aberrations, SCEs, and micronuclei, have been consistently
found in in vivo animal studies  (see Table 4.3). Of particular interest
are the results of a study by Tice et al. (1982)  in which significant
increases in SCEs in bone marrow cells were observed  in mice exposed co
28 ppm benzene for 4 h.

-------
68     Section  4
                               Tabfc4J  la tiro gtsMtoKklry stadies of
End pout/route

Test system
GcMmaudo
ChromoMa
Results
as (no data)
ml effects
References

      Chromosome aberrations
        Inhalation              Mouse bone marrow
        Oral


        Intrapentoneal


        Subcutaneous

        Dermal

     Micron uclei
        Inhalation


        Oral
       Intrapentoneal


       Subcutaneous
       Dermal
     SCE
       IntraperitoneaJ

       Oral
       Dermal
  Rat bone marrow

  Mouse bone marrow

  Chinese hamster bone marrow
  Mouse bone marrow

  Rat bone marrow

 Rat bone marrow
 Rabbit bone marrow
 No data
 Mouse lymphocytes
 Mouse bone marrow
 Rat lymphocytes
 Mouse bone marrow
 Mouse circulating erythracytes

 Chinese hamster bone marrow

 Mouse bone marrow


 Mouse bone marrow
 No data



 Mouse bone marrow

 Mouse lymphocytes
 Rat lymphocytes

Pregnant mouse bone marrow
Fetal mouse liver

No data
No data
 +     Zhurkov et al.  1983.
        Ticect al. 1982.
        Tice et al. 1980
 +     Styles and Richardson 1984.
        Anderson and Richardson 1981

 +     Siou et al. 1981.
        Meyne and Legator 1980
 -I-     Skmetal  1981

 +     Pavlenko et al.  1979,
        Meyne and Legator 1980
 +     Anderson and Richardson 1981

 +     Philip and Krogh Jensen 1970
 +     Kissling and Speck 1972
 +      Erexson et al. 1986.
 +      Toft et al. 1982.
 +      Erexson et al. 1986
 +      Anonymous  1986,
        Gad-el Kanm et al.  1986.
        Fel'dt 1985.
        Harper et al. 1984.
        Siou et al. 1981.
        Meyne and Legator  1980.
        Kite et aL 1980.
        Barak et al. 1983
 +      Baraleet al.  1985,
        Choy et al. 1985.
        Siouetal. 1981

 +      Anonymous 1986,
        Gad-el Kanm et al. 1986.
        Meyne and Legator 1980

 +      Diaz etaL 1980
+     Tice et al. 1982.
       Tice et al. 1980
+     Erexson et al. 1986
+     Erexson et al. 1986

+     Sharma et al. 1985
+     Sharma et aL 1985

-------
                                                                     ToxicoLogical Data
                                                                            69
                                     Table O  (coadaoed)
       End point
         Test system
                                                        Results
         References
Dominant lethal

  Inhalation
  Oral
  Intraperitoneal
Sperm head abnormality
  Inhalation
  Oral
  Intrapentoneal

Cell cycle progression
  Inhalation

  Subcutaneous

  Oral
  Dermal

RNA synthesis inhibition
  Subcutaneous

  Inhalation
  Oral
  Dermal

DNA synthesis inhibition
  Subcutaneous
  Inhalation
  Oral
  Dermal

DNA binding
  Inhalation
  Subcutaneous
  Oral
  Dermal
                                  Chromosomal effects (con't)
No data
Mouse germ cells
Mouse spermatogonia
Rat germ cells
                                         Other effects
No data
No data
Mouse (spermatogonia treated)
Mouse bone marrow

Rat bone marrow
Rat lymphocytes
No data
No data
Rat liver mitochondria
Rabbit bone marrow
No data
No data
No data
Rabbit bone marrow
No data
No data
No data
Rat liver
Rat liver mitochondria
No data
No data
Fel'dt I98S.
Pavlenko et al.  1979
Lyon 1976 (as reported
in Dean 1978)
Topham 1980
Tice et al. 1982,
Tice et al. 1980
Irons et al. 1979
Irons et al. 1979
Kalf et al. 1982
Kisshng and Speck 1972
Kisshng and Speck 1972
 Lutz and Schlatter 1977
 Kalf et al. 1982  .

-------
70   Section 4

     Dominant lethal effects have not been conclusively demonstrated in
animals following benzene administration. Pavlenko et al. (1979)
reported dominant lethal induction in mouse spermatogonia following an
intraperitoneal injection of 3,000 mg/kg benzene.  Postimplantation loss
in treated mice was approximately double that of controls. Fel'dt (1985)
found no significant dominant lethal effect in random-bred male mice
following oral administration of up to 320 mg/kg benzene. Although the
dose in the latter study is an order of magnitude lower than that in the
first study, the two reports cannot be directly compared, because of a
different route of administration and unknown genetic constitution of
the mice used. An intraperitoneal injection of 440 mg/kg benzene induced
no dominant lethality in male rats (Lyon 1976, as reported in Dean
1978). Benzene (or a metabolite) was shown by Topham (1980)  to reach the
germ cells when he obtained positive results in a sperm-head morphology
assay in CBCFl mice. A dose range of 0.1 to 1.0 mL/kg/day for 5 days was
used with the sperm sampled 5 weeks after the last dose. The maximum
induction of sperm-head abnormalities occurred at 0.6 mL/kg/day, while
the highest dose tested was lethal.

4.3.5.4  In vitro

     The potential genetic toxicity of benzene has been examined in a
number of cellular and subcellular systems, including isolated
mitoplasts, bacteria, yeast, and mammalian cells in culture  (see
Table 4.4). End points considered in these studies were DNA binding, DNA
synthesis, DNA repair, gene mutations, and effects at the chromosome
level such as chromosome aberrations, aneuploidy,  SCEs, and micronuclei
formation.

4.3.5.5  General discussion

     Both structural and numerical chromosome aberrations have been found
consistently in bone marrow cells of persons occupationally exposed to
benzene. The conclusion, based on human epidemiological studies, that
benzene is a human clastogen is well supported by in vivo animal studies
and in vitro cell culture and subcellular studies. Virtually all studies
that looked for effects at the chromosomal level were positive when the
ability to metabolize benzene was present. These experimental results
are consistent with the chromosomal damage seen in exposed humans. The
leukemia observed in some benzene-exposed persons may result from the
appearance of a clone of chromosomally abnormal cells in the bone
marrow. With respect to genetic effects, no safe human exposure level
can be determined from available epidemiological data. Significant
increases in SCEs were produced in bone marrow cells of mice at 28 ppm
benzene, the lowest level tested in that study. The significance of SCEs
is unknown, but their production by a chemical is generally considered
to indicate a genotoxic potential. These exposures were via inhalation
and,  based on animal studies, greater effects may result following oral
than inhalational exposure to comparable levels of benzene.  Data
presented in the genotoxicity section and elsewhere in this review show
that benzene metabolites are the active entities;  different metabolites
are possibly responsible for different genetic effects. Differences in
metabolic capability are likely responsible for some of the variations
in response to benzene seen in different test systems.

-------
                                                                  lexicological  Data    71
                      Table 4.4. la vitro geaotoxicity studies of bcaxcM
      End pout
         Test system
 Results"
                  References
Ames test
               One mutations
Salmonella lyphimurium
Azaguanine resistance    Salmonella typhimunum
Histidme reversion
Methionine suppressors

Stamen hair test
Sex-linked  recessive
lethal
TKtest
TK. ouabain,
HGPRT loci
Bacillus subttlu

Aspergillus nidulani

Tradescanna

Drosophila melanogaster


Mouse LSI78Y cells

Total of 15 studies using
various human, mouse, and
Chinese hamster cells
Mixed •
           De Flora et al  1984.
           Shimizu et al  1983.
           Hermann 1981.
           Shahin and Fourmer 1978.
           CSSTT Study 1985*

           Seixas et al  1982.
           Kadenetal  1979

           Tanooka 1977

           Crebelli et al 1986

           Schairer et al.  1979

           Kale and Baum 1983
Oberly et al. 1984
CSSTT study 1985
                           *
Chromosome
aberrations
SCE
Micronuclei
Recombinatioa
Heritable
tramlocation
Human lymphocytes
Total of 8 studies using
Chinese hamster or human
cells
Barley, onion, and broad
bean root tips
Chinese hamster ovary and V79
cells and rat RL4 cells
Human lymphocytes
Chinese hamster ovary cells
Drosophila melanogaster
Spermatocytes
Spennatogoma
Drosophila melanogaster
 Mixed     Gerner-Smidt and
           Fnednch 1978,
           Monmoto 1976,
           Koizumi et al. 1974
 Mixed     CSSTT study I98S*
   +      Zhang and Dong 1982

  -/-     CSSTT study 1985*

 Mixed    Monmoto 1983,
           Gerner-Smidt and
           Fnednch 1978.
           Monmoto and Wolff 1980.
           Monmoto et al. 1983.
           Erexson et  al IMS,

  -/-     Douglas etal. 198S
           Kale and Baum 1983
                                                                Kale and Baum 1983

-------
72      Section  <4
                                           Table 4.4 (condimcd)
             End point
          Test system
Results
                                                                                References
       DNA binding
      ONA breaks
      Unscheduled
      DNA synthesis
      Differential killing
      DNA synthesis
      inhibition
                 Other effects
 Rabbit bone marrow miioplasts       +
 Rat liver miioplasts                 -t-
 Rat hepatocytes                    -

 Chinese hamster V79 cells           -
 Chinese hamster ovary cells        Mixed

 Mouse L5178 Y cells

 Rat hepatocytes                     -
HeLa cells
EschencHia coll                   Mixed

Bacillus subiilu                   Mixed

Human leukocytes                   +
HeLa cells                       Mixed
           Rushmore et al  1984
           Rushmore et al  1984
           Sma ei al  1983.
           Bradley 1985
           Swenbergetal 1976
           Douglas et al  1985.
           Lakhamsky and
           Hendncks 1985
           Pellack-Walker and
           Blumer 1986
           Probst et al. 1981.
           Williams et  al. 1985.
           Probst and Hill 1985.
           Glauert et al 1985''
           Barrett 1985
           Deflora et al. 1984,
           McCarrollet al.  I981b
          Tanooka 1977.
           McCarroll el al.  198la
           Dobashi 1974
          Dobashi 1974.
          Painter and  Howard 1982
         "With/without an exogenous metabolic activation system.
          Progress in Mutation Research 5 1985. Ashby J, de Series FJ. Draper M. Ishidate M Jr. Mar-
     golin BH. Matter BE. Shelby MD. eds Evaluation of Short-Term Tests for Carcinogens  Report of
     the International Program on Chemical Safety Collaborative Study on in vitro Assays. Elsevier.
     Amsterdam.
         'The CSSTT working group disagreed over data analysis and therefore called the results incon-
     clusive.
          Weak positive result

-------
                                                 lexicological Data   73

4.3.6  CareInogen icIty

4.3.6.1  Overview

     IARC concluded Chat there was sufficient evidence that benzene is
carcinogenic (leukemogenie) in humans by inhalation and there was
limited evidence that benzene is carcinogenic in animals.  EPA has
similarly listed benzene as a human carcinogen, Group A, using the EPA
Cancer Risk Assessment Guidelines. There is, however, still controversy
concerning dose-response relationships in humans.

     The Gene-Tox Carcinogenesis Panel categorizes benzene as having
sufficient positive evidence for carcinogenicity in animal studies, and
the NTP concluded that there was clear evidence of carcinogenicity of
benzene for the strains of rats and mice tested in their 2-year
oral/gavage bioassay program.
     Data are insufficient to validate carcinogenic potential via
ingestion in humans or by the dermal route in humans or animals.  It is
reasonable to assume, however, that benzene could cause cancer in humans
if ingested in sufficient quantities. The risk of cancer after dermal
exposure is probably less than by other routes, since absorption through
the skin is low.

4.3.6.2  Inhalation
     Human.  There are many epidemiological and case studies that
correlate benzene exposure with leukemia in humans. Although some of
these studies suggest that low levels of benzene may be carcinogenic,
this is not universally accepted. "Controversy involves the actual
levels of benzene exposure, small number of statistics, and the 'one-hit
concept' implying linear response and no threshold" (Cronkite 1986).  The
duration and levels of benzene exposure in such retrospective studies
are usually deficient or cannot be obtained (Andrews and Snyder 1986),
and cause-effect relationships are complicated by exposure to other
chemicals in addition to benzene (Andrews and Snyder 1986, Dean 1978).
     Pertinent information for seven selected case studies correlating
benzene exposure with carcinogenesis is presented in Table 4.5. In 1976,
OSHA and NIOSH reached the conclusion that benzene was a leukemogen in
humans (EPA 1985a, NIOSH 1977a), and other EPA reviews concur (EPA
1984a, EPA 1986). The 1982 U.K. Health and Safety Executive accepts the
relationship between exposure to benzene in higher concentration ranges
and acute myelogenous leukemia (HSE 1982, as reported in Dean 1985). The
1982 IARC Monograph on cancer summarizes: "It is established that
exposure to commercial benzene or benzene containing mixtures can cause
damage to the hematopoietic system including pancytopenia. The
relationship between benzene exposure and the development of acute
myelogenous leukemia had been established in epidemiologic studies.
Reports linking exposure to benzene with other malignancies were
considered to be inadequate for evaluation. There is sufficient evidence
that benzene is carcinogenic in man" (IARC 1982).
     The studies that were used for quantitative assessment by EPA
(1986) are Ott et al. (1978), Rinsky et al. (1981), and Uong et al.
(1983, as reported in (EPA 1986). Even the better studies, however, did

-------
74    Section
                      Table 4.5 Cue studies of workers occupadonaUy exposed to benzene
Group studied
44 Pancytopemc patients
exposed to benzene in
adhesives
42 Leukemia patients and
21 patients with other
malignancies. 47 were shoe
workers, the remainder in
other occupations using
benzene solvents



6 of 94 Hodgkin's patients
who had been exposed to
benzene adhesives
A 35-year-old man who had
used benzene 8 yean earlier
as a paint solvent
6 Leukemia patients in
different occupations all
using benzene solvents
A 67-year-old man exposed to
benzene in a rubber
hydrochlonde plant
A 51 -year-old chemical worker
exposed to benzene 1 5 yean
earlier
Exposure level/
duration
1 50-650 ppm/
4 mo- 15 years
Not given




150-2 10 ppm
1-28 yean
200-1640
mg/m3/l8
months
Levels
unknown/
1-20 yean
16 ppm/
14 year
<2 ppm/
18 mo
Condition
observed
Leukemia
Myeloid
metaplasia
Leukemia
Multiple myeloma
Myeloblastic
leukemia
Acute
erythroleukemia
Pre leukemia
Malignant
lymphoma
Paroxymal
nocturnal
hematuna
Lung cancer (all
heavy smokers)
Hodgkin's
disease
Subacute
granulocytic
leukemia
Hemocvtoblastic
leukemia
Acute
myeloblastic
leukemia
Acute
myelogenous
leukemia
Author
call'
PC
•
DC
PC
•
•
•
PC
•

•
PC
DC
DC
DC
1981
PC
References
Aksoy and Erdem
1978
Aksoy 1980




Aksoy et al.
1974b
Sellyei and
Kelemen 1971
Vigliam and
Saita 1964
Rinsky et al.
Ott et al.
1978
         DC -  Direct correlation.
         PC - Possible correlation.
         •  - Author made no claim.

-------
                                                 ToxicoLogical Daca   75

not have complete exposure information. Ott et al. (1978), while finding
an excess of leukemia deaths, points out that the varied work histories
and the lack of medical histories make a retrospective assessment of the
possible relationship to benzene exposure Judgmental. EPA used the Ott
study in combination with the other studies for risk estimates,  but less
emphasis was placed on it, because of a lack of a clear-cut dose-
response relationship (EPA 1986).
     The initial study by Infante, later updated by Rinsky, relied on a
1946 Industrial Commission of Ohio survey of one department and on
company surveys between 1963 and 1974 to estimate exposure levels
(Infante et al. 1977). "There was a paucity of information regarding
atmospheric levels of benzene to which workers may have been exposed,
but the available data indicated that average benzene levels generally
ranged from less than 10 ppm to 100 ppm" (White et al. 1980). In the
discussion in the IARC Monograph (1982) concerning this work, it was
stated that "the methods employed in the 1940s for measuring benzene
concentration in air, while reasonably accurate, were relatively less
sensitive than those available today." In fact, in laboratory studies,
one type of detector kit used read 40% low (Hay 1964, as reported in
Rinsky et al. 1981), and a detector tube used as late as the 1960s was
found to have an accuracy of ±50% (Ash and Lynch 1971, as reported in
Rinsky et al. 1981).
     In a follow-up to the Infante study (Rinsky et al. 1981),
correspondence, memoranda, and records of air-sampling measurements from
several sources taken at various times between 1947 and 1977 were cited
as documentation of exposure levels at location 1 of the study. "There
is limited data for location 2 and no data available prior to 1946 when
an exhaust system was installed. One might infer that prior  to 1946 the
presses were not enclosed and therefore benzene emissions were higher.
We believe that benzene exposures at location 2 were similar to location
1. Our analysis of the available data lead us to conclude that for the
most part employees' 8 h TWA exposures (which ranged from 100 ppm in
1941 to 10 ppm in 1969) were within the recommended limits in effect at
the time. However ... on certain days the 8 h TWA was probably
exceeded" (Rinsky et al. 1981).
     The IARC Working Group "accepted the central conclusion of Infante
et al. and Rinsky et al. that excessive mortality from myelogenous
leukemia had occurred among workers with occupational exposure to
benzene that was generally within accepted limits. However the possible
contribution of the occasional excursions in exposure and of the
employment of some workers in other areas of the plant must  be noted;
and in the opinion of the Working Group, these  factors may have made
some contribution to  the observed excess in mortality from leukemia"
(IARC 1982).
     Van Raalte and Grasso (1982) did an extensive critical  review of  a
number of studies reporting positive results,  including  Infante  et al.
(1977) and Rinsky et  al.  (1981). This review takes issue with sampling
techniques, exposure  determinations, mortality  standards,  and other
aspects of experimental design or methodology.  Goldstein (1985)
emphasizes that in dealing with  a low-incidence phenomenon it  is

-------
 76    Section 4

 difficult Co utilize general industrial  hygiene measurements  as  a  means
 of  typifying individual exposure.  Van Raalte  and Grasso  (1982) and
 Goldstein (1985)  concluded that the  cause-effect relationship between
 benzene and leukemia is sufficiently clear, but Van Raalte and Grasso
 assert that there are few data from  which dose-effect or dose-response
 relationships can be established.  They claim  that  there  is no evidence
 for a leukemogenic action of benzene at  low concentrations, and,
 although an association has been suggested for lymphatic leukemia,
 Hodgkin's disease,  and non-Hodgkin's lymphomas, Van Raalte and Grasso
 consider this to  be questionable.

      On the other hand,  White et al.  (1980),  reviewing arguments in
 OSHA's case to lower occupational  benzene standards from 10 to 1 ppm,
 examined a study  by Thorpe (1974)  in which no correlation between
 benzene exposure  and cancer was found. They attributed the lack of
 correlation to methodologic deficiencies. They further summarized  that
 OSHA's earlier conclusion that no  threshold could be established for
 benzene is supported by the consensus of the  scientific community.
      Conditions observed and the investigators' assessment of
 correlation from  a  few of the better-known epidemiological studies are
 summarized in Table 4.6.

      Since the quantitative assessment by the EPA,  Rinsky et  al. (1987)
 have  published a  risk assessment based on an update of the previous
 Infante and Rinsky  studies of a cohort of 1,165 rubber workers. In order
 to  reduce  the uncertainties posed  by estimates of group exposures,
 individual work histories were compiled and cumulative exposures were
 estimated  for each  employee in the cohort based on the available past
 industrial hygiene  measurements, which, as discussed above, were
 limited.  In some  cases,  a single measured exposure served as  an index of
 exposure  for a number of years.  (For details of these measurements, see
 Rinsky et  al.  1981.)  Standardized  mortality ratios (SMRs) were
 determined for leukemia  by four cumulative exposure categories.

         Cumulative exposure
              (ppm-years)	SMR    95% Confidence interval
< 40
40-200
200-400
> 400
Total
109
322
1,186
6,637
337
12-394
36-1,165
133-4,285
1,334-19,393
154-641
     The questions discussed above remain concerning the precise levels
of benzene Co which workers were exposed. Additionally, there is no
means of assessing the impact of the manner in which workers were
exposed: I.e., short-term high-level exposure, long-term low-level
exposure, exposure at a constant level with occasional high-level
exposure, or only occasional high-level exposure, all of which could
produce Che same cumulative dose. However, the trend demonstrated is a
marked, progressive increase in SMR with increasing cumulative dose. The

-------
                                                              Toxicological  Data    77
                   Table 4.6. Epidemiological studies of worfccn exposed to
Group studied
Mortality study of rubber
Corkers exposed to benzene
between 1940 and 1949






Follow up mortality study
of rubber workers exposed
to benzene between 1 940 and
1949




Mortality study update of
rubber workers exposed to
benzene between 1 940 and 1 949



Mortality study of 594
workers employed at a
chemical company between
1940 and 1973


Mortality study of 4.602
chemical workers employed
between 1946 and 197S





Mortality study update of
594 previously studied and
362 additional chemical
employees potentially
exposed to benzene between
Exposure level/
duration
Within legal
limits of
time, i e ,
100 to 10 ppm/up
to 10 years or
more



Within legal
limits of the
time, i e ,
100 to 10 ppm/up
to 10 years
or more


Redefined
from
<40 ppm • yean
to >400 ppm years


<0 1-35 5
ppm estimated
TWA/up to 34
years


TWA of from
<1 to >50 ppm
with peaks to
>100 ppm/6 month
to 29 years



1-30 ppm/
duration
not well
documented

Condition
observed
Leukemia (all
types)
Lymphatic
hematopoietic
cancer
Myeloid
leukemia
Monocytic
leukemia
Malignant
neoplasias of
the lymphatic
and hematopoietic
tissue
Leukemia
(myelogenous)
(monocytic)
Lymphatic and
hematopoietic
neoplasms
Leukemia
Multiple
• myeloma
Leukemia
Acute
myelogenous
leukemia
Myeloblastic
leukemia
Leukemia and
Aleukemia
Lymphatic and
hemoreticular
cancer
Non-Hodgkin's
lymphopoietic
cancer
All leukemia

Acute
myelogenous
leukemia
Author
call"
DC

DC


DC

DC

DC




DC


DC


DC
DC

PC
PC


PC

DC

PC


PC


•

DC


SMR References
506 Infante et
at 1977,
260 Infante
1978





330 Rinsky et
al 1981



560


227 Rinsky et
al. 1987

337
409

* Ott et al.
• 1978


•

97-275 Wong et al.
1983. as
91-175 reported in
EPA 1986

108-165


158-214 Bondetal.
1986
444


1938 and 1970

-------
78
Section  <4
                                            Table 4.6 (continued)
Group studied
Mortality study of workers
employed at a Texas
refinery between 1952 and 1981



Mortality study of
commercial pressmen
members of Local 51 who
died between 1950 and 1981


Exposure level/
duration
<1 ppm/up to
27 years




Levels not
given — benzene
and other
solvents/
duration not
well documented
Condition
observed
Stomach cancer
Cancer of
buccal cavity
Pharynx cancer
Larynx cancer
(No leukemia)
Lymphatic
cancer
Hematopoietic
cancer
Non-Hodgkin's
lyrnphoma
Author
call" SMR References
• • Tsai et al
• • 1983

• •
• •

• • Zoloth et
al 1986
• •

• •

        9 DC  — Direct correlation.
         PC =• Possible correlation.
         •  = Author made no claim

-------
                                                 Toxicological Data   79

95% confidence intervals are quite wide, particularly for the higher
doses, which is to be expected with only nine leukemia deaths. There was
no apparent pattern for these deaths with regard to latency, which
ranged from under 5 years to more than 30 years. There was also a
statistically significant excess of death from multiple myeloma.
                    •    f
     In addition, numerous other studies have evaluated the relationship
between benzene exposure and cancer but without attempting to quantify
the relationship. Alderson and Rushton (1982) found a deficit of
leukemia and no excess of myeloid leukemia in a mortality study of
35,000 workers at eight oil refineries in the United Kingdom. There was
no monitoring for benzene exposure, but the authors say the average
benzene exposure is "likely to have been greater than for the population
as a whole."
     A case-control study was conducted as an extension of this
investigation (Rushton and Alderson 1981). Based on job history,  each
worker was allocated to a benzene exposure level of "low," "medium," or
"high," and deaths from leukemia were analyzed. The risk for the workers
with medium or high exposure, relative to the risk for workers with low
exposure, approached significance (? - 0.05) when length of service was
taken into account. The authors state that if there were an increased
risk of leukemia associated with benzene exposure, only a very small
proportion of the refinery work force would be affected. Alderson and
Rushton (1982) point out that the eight refineries have a quite
different range of plants and processes; that the workers have been
employed for varying lengths of time and at varying job assignments; and
that epidemiological studies at oil refineries are likely to be beset
with problems of mixed exposures and small numbers of men who have
worked at specific plants.
     In a follow-up to a study by McMichael et al. (1975), Vilcosky
(1984) did not find any correlation between benzene and any of the
cancers reported. He points out, however, that "exposure" should be
reinterpreted in this study as "potential exposure" and that all of the
leukemia cases in the earlier study had died before the beginning of the
Wilcosky study.
     Decoufle et al. (1983) found a significant increase of leukemia
deaths in a study of chemical workers, but made no attempt to correlate
exposure levels beyond saying that it was from fugitive emissions
peculiar to the technology, quality control, and maintenance procedures.
They also observed that some employees' personal habits such as cleaning
hands, tools, and clothes in benzene and siphoning benzene for home use
contributed to their exposure. Although not reported in other studies,
this may not be a unique occurrence.
     Aksoy et al.  (1974a) reported a leukemia  incidence during 1967 to
1973 of 13/100,000 among 28,500 Istanbul shoe workers exposed to 150 to
650 ppm benzene for 4 months to 15 years. That is significantly higher
than the estimated 6/100,000 of the general population. After the
phaseout of benzene in 1969, the number of leukemias decreased, and none
were reported in the subsequent 3 years  (Aksoy 1980). IARC  (1982)
comments that "the estimates are limited by the study design
characteristics and by  the uncertainties about the way  in which cases
were ascertained and how many of the study population were exposed  and

-------
80   Section 4

how many unexposed." EPA  (1986) called the "exposure information so
imprecise that they are not suitable for quantitative assessment."

     Yin et al. (1987a) did a retrospective cohort study of 28,000
benzene workers in China,- all of whom had worked in various factories
for at least half a year  between 1972 and 1981; however, exposure and
employment duration were  not necessarily limited to those years. Thirty
cases of leukemia (23 acute, 7 chronic) with a mean latency of 11.4
years (0.8 to 49.5 years) were found in the benzene cohort, as opposed
to 4 in a matched control cohort. Twenty-five of the leukemic workers
had already died. Information on exposure levels was collected from
company records, but there is no indication of the extent of these
records with the exception of a note that three levels were based on
only one measurement. Mean benzene levels to which workers developing
leukemia were exposed ranged from 10 to 1,000 mg/m3 (-3 to 330 ppm),
with the majority falling between 50 and 500 mg/m3 (16 to 160 ppm). It
should be noted that the  exposure ranges from which the means were
derived were rather wide, indicating the possibility of at least
occasional high exposures. Only four upper-level measurements were less
than 10 ppm, while half of the remaining cases were between 10 and
100 ppm, and the other half were between 100 and 2,000 ppm. The authors
observed that the cumulative mortality of leukemia was in proportion to
the duration of the exposure to benzene up to 20 years and then leveled
off.

     The same group reported that between 1979 and 1981 Chinese workers
using benzene or benzene-containing mixtures were examined, and nine
cases of leukemia were found. (Presumably, some of these may have been
reported in the earlier study discussed above.) Although one worker was
exposed for only 2 years, the others were exposed for between 7 and 25
years.  No estimate of exposure levels was given for the leukemia cases,
but exposure estimates for aplastic anemia cases found in the same study
were 93 to 1156 mg/m3 (-30 to 360 ppm) (Yin et al. 1987b).
     In order to give the reader a slightly broader view of the
literature than is possible within the scope of this report, some
primary sources not included elsewhere in this section are cited in
Table 4.7 along with the  review in which they were found. The reader
should not assume that these were the only sources cited by the reviewer
or the sole basis for the reviewer comments included. With"the exception
of Mallory eC al. (1939,  as reported in IARC 1982) and Gallinelli  (1966,
as reported in Goldstein  1977), the human studies are of persons exposed
or potentially occupationally exposed to varying levels of benzene for
varying lengths of time.

     Animal.  Recent studies have shown benzene to be carcinogenic in
animals by inhalation although there is not universal acceptance of all
results, primarily because of the difficulty of developing reliable
animal models for benzene-induced leukemia.
     The Goldstein et al. (1982a) study, using lifetime inhalation
exposure, was the first to report leukemia in test animals. Although it
is not statistically significant compared with controls, the authors
point out that the granulocytic leukemia found is extremely rare in
Sprague-Dawley rats, even after treatment with known leukemogenic
agents.  They suggest the  importance of the work in terms of a potential

-------
                                                                 lexicological Data     81
           Table 4.7.  Carcraogeokity review stadia of occnpatkwally exposed workers
   Reviewer
   Work cited
 Conditions reported
      Reviewer's comments
R  Snyder 1984
Kalf et al
1987
Infante and
White 1983
Cole and
Merletti 1980
 I ARC 1982
McMichael
etal 1975.
Monson and
Nakano 1976
Vighani 1976,
Arpet al  1983
Browning 1965
                 Vighant and
                 Form 1976
Tabershaw and
Lamm 1977,
Zenz  1978
 Aksoy 1985a      LeNoir 1897

                  Bousser et al
                  1947

                  Vianna and
                  Polan 1979
 Torres et aL
 1970

 Delore and
 Borgamo 1928
Lymphatic leukemia.
myelogenous
leukemia.
lymphosarcoma
Myelogenous
leukemia
Acute myeloid
leukemia.
subacute myeloid
leukemia.
chronic myeloid
leukemia,
lymphatic  leukemia,
erythroleukemia


Acute leukemia,
chronic myeloid
leukemia.
chronic lymphatic
leukemia

Leukemia
                   Leukemia

                   Lymphosarcoma
 Malignant
 lymphoma

 IgG myeloma
 Lymphoblastic
 leukemia
Most observers agree that the
lowest air levels of benzene
demonstrated to produce decreases
in any circulating blood
cells in humans are in the range
of 40 to 50 ppm over a penod of
lime.  Until the mechanisms of
action of benzene are determined
and used in the preparation of
nsk estimates, debates over
acceptable levels of human
exposure will continue

Benzene is a carcinogen
associated with increased
incidence of myelogenous leukemia
in humans

Epidemiologic studies of workers
have been too insensitive to
determine nsk of death from
cell types of leukemia that may
have nsk ratios of leu than
5 0. Case studies suggest an
association of chronic leukemia.
including lymphatic leukemia and
benzene
There is sufficient evidence
that benzene is carcinogenic to
hematopoietic tissues in man
(leukemia)

There seems to be sufficient
data to incriminate benzene as a
potent carcinogen causing
leukemia, malignant lympnoma,
multiple myeloma, and lung
cancer. Genetic factors may
have a role in the development
of condition
 It is established that human
 exposure to commercial benzene
 or benzene-containing products
 can cause damage to the hematop-
 oietic system, including
 pancytopema. The  relationship
 between benzene exposure and the

-------
82
Section
                                      Table 4.7 (coatfamed)
      Reviewer
                       Work cited
                                               Conditions reported
                           Reviewer's commenu
                     Hunter 1939
    White et aL
      1980
                     Mallory
                     et al. 1939

                     DeGowm 1963

                     Tareeff et al.
                     1963

                     Goguel et al.
                     1967
                            Ludwig and
                            Wenhermann
                            1962
                            Galavotti and
                            Troisi 1950,
                            Nissen and
                            Soeborg
                            Ohlsen 1953,
                            DiGuhelmo and
                            lannaccone
                            1958.
                            Rozman et al.
                            1968.
                            Byron et aL
                            1969.
                            Forni ftfld Moroo
                            1969.
                            Girard and
                            Revol
                            1970


                            Ishimani et
                            al. 1971
                                                            development of acute myelogenous
                                                            leukemia has been established in
                                                            epidemiological studies.
                                                            Reports linking benzene to other
                                                            malignancies were considered to
                                                            be inadequate for evaluation.
                                                            There is sufficient evidence
                                                            that benzene is carcinogenic in
                                                            man
Lymphoblastic
leukemia (non-
occupational—
12-year-old boy
who frequently
used paint remover)

Acute myeloblastic
leukemia

Myeloid leukemia

Leukemia


Myeloid leukemia,
chronic lymphoid
leukemia,
acute leukemia,
eryihroleukemia

Myeloid leukemia
                                       Erythromyelosu
Acute leukemia,
chronic lymphoid
leukemia,
myeloid leukemia

Leukemia

-------
                                   Table 4.7 (continued)
                                                                  Tax Leo logical  Data     83
   Reviewer
    Work cited
Conditions reported
                                                               Reviewer's comments
Goldstein 1977
 Emile-Well 1932.
 Oldfelt and
 Knutson  1948.
 Curletto  and
 Ciconah  1962.
 Inceman  and
 Tangun 1969,
 Kinoshita et
 al I96S,
 Tzanck et al
 1937.
 Zini  and
 Alessandn
 1967,
Gallmelli
 1966
                Appuhn and
                Goldeck I9S7.
                Kohli et al.
                1967
                                   Acute myelogenous
                                   leukemia
                  (Nonoccupational—
                   16-yr-old an
                  student)

                  Erythroleukemia
                     Occupational exposure to benzene
                     appears causally related to
                     acute myelogenous leukemia and
                     its acute variants  The
                     question of whether leukemogene-
                     sis occurs only after high-dose
                     benzene exposure leading to
                     significant bone marrow damage
                     remains unanswered

-------
 84    Section  4

 model for  the study  of benzene-Induced  leukemia. The same group  (C. A.
 Snyder et  al. 1984)  suggest a possible  causal relationship between
 benzene and the  incidence  of certain  tumors that rarely occur
 spontaneously.

      Maltoni  et  al.  (1982c,d; 1983; 1985) have reported inducing Zyrabal
 gland carcinoma,  for which there  is no  human counterpart, and other
 tumors by  inhalation.  The  EPA Gene-Tox  Carcinogenesis Panel (Nesnow et
 al.  1986),  in an evaluation of  the Haltoni et al. (1982d) and Maltoni et
 al.  (1983)  studies,  called results of the inhalation experiments
 inconclusive.

      A major  continuing study (Cronkite et al. 1984, 1985; Cronkite
 1986)  provides a basis on  which a reproducible model may be built. In
 this  study  mice  were exposed to 300 ppm benzene by inhalation 6 h/day,
 5 days/week for  16 weeks.  This  exposure regimen was selected because the
 authors  thought  it most closely paralleled likely human exposure. Human
 epidemiological  studies in the  literature reported that occupational!/
 exposed persons  were exposed for about  15% of their life span; 16 weeks
 represents  -15%  of the life span for mice. Although precise occupational
 exposures were not known,  most  studies  indicated that workers had been
 exposed  to  up to  250 to 300 ppm during at least part of a working day;
 thus  300 ppm was  chosen. The C57B1/6 and CBA/Ca mouse strains were
 chosen for  these  studies because of their respective susceptibilities to
 ionizing radiation-induced thymic lymphoma and acute myeloblastic
 leukemia (AML).   The  strains are also recognized for their low
 spontaneous rates of AML,  the disease most frequently associated with
 benzene exposure  in  humans.

     Cronkite et  al.  (1984)  reported a highly significant increase in
 leukemia in C57B1/6  mice after  exposure according to the above regimen.
 In a continuation of that  study (Cronkite et al.  1985), a definite
 pattern for lymphoma appearance and mortality was observed.  A first wave
 of lymphoma commenced  at about  150 days after exposure. The mice began
 to die at 330 days,  and mortality increased through 390 days,  at which
 time it leveled off. A second wave of lymphoma and solid tumors began
 420 days after exposure, and mortality did not increase again until 570
 days after exposure. This  is a  "significantly different pattern for the
 appearance of lymphoma and solid tumors than that after lifetime
 exposure (i.e. "lifetime exposure" as reported by C. A. Snyder et al.
 1980). This suggests that  continuous exposure either suppresses the
 incidence of lymphoma  or shortens the life span of the mice so that
 lymphoma cannot be observed."

     In 1986,  Cronkite  reported that 100 or 300 ppm benzene is
 carcinogenic  (leukemogenic)  in  both male and female CBA/Ca mice.
 However, no experimental details for the 100-ppm effect are given in the
 report.

     Inhalation  is the  most common route of exposure to benzene for
humans, and the successful induction of cancer in animals by this route
 is an  important step in understanding the most basic questions
 concerning benzene and human health. Some of these are the mechanisms of
cancer development,  the role of genetic factors in the development of
cancer, and, most important from a regulatory standpoint, dose-response
 relationships.

-------
                                                 ToxLcologLcal Data   85

     Rather than describing all experiments in the text, Tables 4.8 and
4.9 summarize examples of the growing body of animal results,  the
oncogenic end points observed, and the investigators' assessment of
correlation. Table 4.8 presents a summary of animal inhalation
experiments that have investigated the potential carcinogenicity of
benzene. Some author comments are given in the footnotes to this table.
Table 4.9 presents essentially the same data but allows a more detailed
examination of tumor type and animal species, while also focusing on the
lowest effective exposure concentrations.

4.3.6.3  Oral
     Human.  There are no studies available.
     Animal.  In recent years, there have been several oral/gavage
studies with positive results. There is some discussion of the relevance
of gavage studies vs other routes by which human exposure is more likely
to occur, especially inhalation and dermal (NTF 1986).
     Investigation of benzene-induced neoplasia in laboratory rodents
has increased (Dean 1985). According to the EPA, recent research has not
shown that benzene is a noncarcinogen, but that the correct model
incorporating exposure route, animal strain, and species has not been
tested (EPA 1986a). Haltoni and Scarnato, exposing rats to benzene by
ingestion, were the first to  demonstrate that benzene is an animal
carcinogen  (Maltoni and Scarnato 1977, as reported in Maltoni 1983;
Maltoni and Scarnato 1979 as  reported in Maltoni 1983).
     Maltoni et al. (1982b, 1983, 1985) have reported inducing Zymbal
gland carcinoma, for which there is no human counterpart, by
oral/gavage. There seems to be general agreement for a  causal role of
benzene in  the formation of these tumors (see IARC 1982, R. Snyder 1984,
Dean 1985). Maltoni et al. (1982b, 1983) also report the induction of
other tumors. The EPA Gene-Tox Carcinogenesis Panel  (Nesnow et al.
1986), in an evaluation of the Maltoni et al. (1982d) and Maltoni et al.
(1983) studies, concurred on  their findings  for the  induction of Zymbal
gland and oral carcinoma by oral/gavage.

     NTP has now completed a  2-year oral/gavage study  of rats and mice
showing positive results  (NTP 1986).  For details of  that study,  see
Tables 4.10 and 4.11.
     Rather than describing all experiments  in  the  text> Tables  4.10 and
4.11 summarize examples of the growing body  of  animal  results,  the
oncogenic end points observed, and the investigators'  assessment of
correlation. Table 4.10 presents a summary  of animal oral/gavage
experiments which have  investigated  the  potential  carcinogenicity  of
benzene.  Some author comments are given  in  the  footnotes  to this table.
Table 4.11  presents essentially  the  same data but  allows  a more  detailed
examination of  tumor  type  and animal  species, while  also  focusing  on  the
lowest effective exposure  concentrations.

-------
86
Section
                        Table 4.8.  Sommmry of inimaJ Inhalmdo. c«rctoog«^eJty expertoerti
Species"
CS7BL/6J mice



AKR/J mice

CD-I mice

SDrat

SDrat

SDrat









C57BL (6 mice)

SD raw




CS7BL [6J mice
(female))
CBa/CA mice (male)
CBA/CA mice (male)
Exposure or
observation tune
488 days '



505 days

Life

86 weeks

116 weeks

118 weeks'









64 weeks

863 days




580 days

800 days
900 days
Effect
Lymphocytic lympboma (thymic
involvement)
Myeloma
Leukemia
Malignant lymphoma

Myelogenous leukemia (acute) [I]f
Myelogenoiu leukemia (chronic) [I]
Zymbal gland carcinoma

Hepatoma

Breeders
Zymbal gland caranoir • I ]
Hepatoma [I]
Mammary carcinoma [1
Offspring-
Zymbal gland carcinoma [I]
Nasal carcinoma [I]
Hepatoma [I]
Leukemia (I)
Mammary carcinoma [I]
Thymic lymphoma [I]
Lymphoma. unspecified [I]
Zymbal gland carcinoma

Liver tumor
Chronic granulocytic leukemia
Mammary carcinoma
Lymphoraa
Solid tumor
Leukemia
Hepatoma
Author
call*
DC
DC
•
•
NC

PC"
PC
DC?

DCf


DC*
PC
•

DC
DC
PC
•
•
DC1
DC
PC

PC
DC
PC
DC
DC
DC
PC
References
C A Snyder
et al. 1980


C A Snyder
etal 1980
Goldstein et al
I982a
Maltoni et al
1982d
Maltoni et al.
1982b

Maltoni et al.
1983


Maltoni et al.
1983



Crookite et al.
1984
C A. Snyder
etal 1984



Cronkite et al.
1985
Cronkite 1986
Cronkite 1986
          "SD - Sprague-Dawley.
          *DC - direct correlation, NC - no correlation, PC - possible correlation, and • - no author call.
          "Gene-Tox Carcinogenens Panel call (Nesoow et al.  1986)- [ + ] - positive, [I] - inconclusive.
          rfThe importance of toe present study is in terms of a potential model for the studv of beniene-induced
      leukemia. Although the authors claim this is the fust case of myelogenous leukemia reported in laboratory
      animals, the incidence of myleoproliferative disease is not significantly higher than controls.  There has. how-
      ever, been no observation of spontaneous myeloproliferative disease in SD rats or CD-I mice.
          'Continuous exposure to 200-300 ppra benzene for 4-7 h daily causes onset of Zymbal gland carcinoma.
      One carcinoma (female) out of 158 male  and 149 female controls.
          •^These results provide the first evidence that benzene causes hepatomas in experimental animals, further
      proof that benzene is a mulbpotential carcinogen.
          'Exposure started at 12 days gestation and continued for 118 weeks. Offspring were thus exposed initially
      as embryos.
          *The results confirm that benzene causes the onset of zymbal gland carcinomas in SO rats, even when
      given by inhalation. Incidence parallels length of treatment.  Nasal carcinoma observed. Appears to cause
      hepatomas and other related dysplastic and hyperplastic lesions.
          'The 88 experimental controls were leukemia free. Death rates due to leukemia-lymphoma for 118 ben-
      zene exposed and 354 recent historical control mice were compared  by a Cox model of survival analysis. Ben-
      zene is a significant variable (P < 0.0001) in predicted lymphoma-leukemia death rale.

-------
                                                                         Toxicological  Data     87
          TaMe 4.9. Carcinogenic-related end points observed ID uiinab exposed to benzene by inhalation"
  End points observed
              AKR/J   C57BL/6J    CD-I     CBA/Ca
  SD rat       mouse      mouse      mouse     mouse
                           References
Lymphocytic lymphoma
Myelogenous leukemia
acute
Myelogenous leukemia
chronic
100/life
                        300/life
                                    300/hfe
300/hfe
C  A Snyder et al  1980
Goldstein et al  1982a

Goldstein et al  I982a
Zymbal gland carcinoma
Hepatoma
Mammary carcinoma
Nasal carcinoma
Thymic lymphoma
Lymphoma unspecified
Liver tumor
Granulocytic leukemia
Solid tumor
Leukemia
100/life
200-300/104
100/hfe
200-300/104
300/16
300/16
100/hfe
100/hfe
300/16

C A Snyder et al. 1984
300/16 Maltom et al. 1982b (rat).
Cronkite 1986 (mouse)
C. A Snyder et al. 1984
Maltom 1983
Cronkite et al. 1984
Cronkite et al 1984
C A Snyder et al 1984
C A Snyder et al 1984
Cronkite et al 1985
300/16 Cronkite 1986
  "Doses are expressed in ppm/h given 4-7 h/day. 5 days/week over a number of weeks or lifetime (e g, 300/99 =
300 ppm given for 99 weeks): exposures shown are the lowest for which author claims possible causal relationship

-------
88
Section
            Table 4.10.  Summary of animal oral/gavage carcinogenicity experiments
Species
strain"
SD rat

SD rat

SDrat








Wistar rat



Swiss mice


F344/N rat


Observation
time* End points observed
84 weeks Carcinoma of oral cavity
Zymbal gland carcinoma
115-157 Glioma
weeks
92 weeks Zymbal gland carcinoma [ + Y
Carcinoma of oral cavity [ + ]
Carcinoma of nasal cavity
Angiosarcoma
Leukemia
Mammary tumor
As above plus:
Hepatoma
Others
100 weeks Zymbal gland carcinoma
Carcinoma of oral cavity
Thymoma
Other hemolymphoreticular neoplasms
100 weeks Zymbal gland carcinoma
Adenocarcinoma pulmonary tumor
Hemolymphoreticular neoplasia
2 years Carcinoma of oral cavity [ + ]
Zymbal gland carcinoma [ + ]
Skin carcinoma (M) [ + ]
Author
callf
DC*
DC
NC'

DC*
DC
DC
PC
DC
DC

DC
PC
DC*
DC
*
DC
DC
DC
DC
DC1
DC
DC
References
Maltoni et al
19826
Maltoni et al.
1982a
Maltoni et al.








Maltoni et al



Maltoni et al.


NTP 1986

^




1983








1985



1985






-------
                                                                   Toxicological Data    89
                                        Table 4.10  (continued)
Species
strain"
B6C3F, mice








Observation
time* End points observed
2 years Zymbal gland carcinoma [ + ]
Malignant lymphoma [ + ]
Alveolar/bronchiolar carcinoma [ + ]
Alveolar/bronchiolar adenoma
Hardenan gland adenoma (M) [ + ]
Preputial gland carcinoma (M) [ + ]
Ovarian granulosa cell tumor (F)
Mammary gland carcinoma (F) [ + ]
Mammary gland carcmosarcoma (F)[ + ]
Author
call'
DC
DC
DC
DC
DC
DC
DC
DC
DC
References
NTP 1986








    "SD = Sprague-Dawley rat.
    * Actual exposure period may have been less.
    CDC = Direct correlation.
     NC = No correlation.
     PC = Possible correlation.
     * = No author call.
    'These results provide the first evidence that benzene causes carcinoma of the oral
 cavity in experimental animals, further proof that benzene is a multipotential
 carcinogen.
    TJnder our experimental conditions, benzene did not seem to produce brain tumors. One
 glioma (male) out of 30 male and 30 female controls.
    7Gene-Tox  Carcmogenesis Panel call (Nesnow et al.  1986): [ + ] = Positive,
 [I] = Inconclusive.
    *Data confirm that benzene causes Zymbal gland carcinoma at two dose levels with dose
 response.  It induces carcinoma of the oral cavity, very unusual in our colony of
 rats: also, liver angiosarcoma very rare in  SD rats.  Benzene increases incidence of
 hemolymphoreticular neoplasms ("leukemias") and mammary carcinoma.
     Benzene by ingestion or inhalation is a multipotential carcinogen  in two strains of
 rats and in mice and precedes a variety of tumors.  Incidence of Zymbal gland
carcinoma and carcinoma of the oral and nasal cavities is affected  by  length of
 inhalation treatment and age of animals.
    'Under the  conditions of these 2-year gavage studies, there was clear evidence of
carcmogenicity of benzene for male and female F334/N rats and male and female B6C3F,
mice.

-------
90    Section
          Table 4.11.  Carcinogenicity-related end points observed in animals exposed
                                 to benzene by garage"

                               SD rat*   Wistar*   F344/N*   Swiss'   B6C3F,'
                                           rat        rat      mouse    mouse

                                               Duration in weeks
       End points observed          52        100        103        78       103

       Zymbal gland carcinoma      SO      500         50       500        50
       Oral carcinoma             250      500         50
       Nasal carcinoma            250
       Leukemia                   50
       Mammary  tumor             50      500
       Hepatoma                  250
       Hemolymphoreticular                 500                 500
       neoplasia
       Skin carcinoma                                 200
       Pulmonary adenocarcinoma                                500
       Malignant  lymphoma                                                25
       Alveolar/bronchial                                                  50
       carcinoma
       Alveolar/bronchial                                                  25
       adenoma
       Hardenan gland                                                    25
       adenoma
       Preputial gland                                                     50
       carcinoma
       Ovarian granulosa                                                  100
       cell tumor
       Mammary  gland                                                    50
       carcinoma
       Mammary  gland                                                   100
       carcinosarcoma
       Hcpatocellular adenoma                                             25
       Hepatocellular carcinoma                                            50

          "Doses  (mg/kg) are the lowest 5-days/week exposures at which author claims
       causal relationship.
          *Maltoni  1983.
          'Maltoni  et al.  1985.
          'NTP 1986.

-------
                                                 lexicological Data   9L

4.3.6.4  Dermal
     Human.  There are no studies available.

     Animal.  There is no evidence that benzene has induced skin tumors,
although not all possible sites were examined in all experiments (IARC
1982). At one time, benzene was used as a solvent vehicle in the study
of carcinogenic compounds. As such it was applied topically as a control
without effect (Burdette and Strong 1941, Bock et al.  1959, Van Duuren
et al. 1969). Interestingly enough, Baldwin et al. (1961) argue against
the carcinogenicity of a compound being tested because "a similar tumor
incidence was observed in the benzene control" (there was no nonbenzene
control). These experiments are not considered adequate to determine
carcinogenic potential.
     Bull et al. (1986) commented that it has been very difficult to
induce tumors in experimental animals with benzene and that the rate of
false-negative responses to chemicals with recognized carcinogenic
activity is quite high when tested on mouse skin.

4.3.6.5  General discussion
     IARC concluded that there was sufficient evidence that benzene is
carcinogenic in humans and that there was limited evidence that benzene
is carcinogenic in animals (IARC 1982). The EPA Gene-Tox Carcinogenesis
Panel (Nesnow et al. 1986) calls benzene a sufficient positive in animal
studies, and NTP concluded that there was clear evidence of
carcinogenicity of benzene for the strains of rats and mice tested in
their 2-year oral/gavage study (NTP 1986). EPA (1988b) has verified the
weight-of-evidence classification for carcinogenicity as EPA category A,
based on a sufficient level of human evidence as  supported by a
sufficient level of animal evidence. There are still questions regarding
both the mechanism of benzene carcinogenesis  (Dean 1985) and the most
appropriate models for developing human  risk  estimates.
     Based primarily on the Crump and Allen analysis of epidemiological
studies  (Crump and Allen 1984) showing an excess  of leukemia cases
(Rinsky  et al. 1981, Ott et al. 1978, Wong 1983), OSHA has lowered the
benzene  level to 1 ppm (OSHA 1987). This analysis uses the linear,
nonthreshold model generally accepted among regulatory bodies  in the
United States for extrapolation of risks. In  the  OSHA hearings, Crump
explained, "Although many dose response  forms could be posited, it was
felt that  the number of leukemias  is too few  (there are only  16
leukemias  in the Rinsky et al., Ott et al., and Wong cohorts  combined)
to permit  discrimination among alternative dose  response models. A
linear model provides an acceptable fit  to all data sets examined  .  .  .
and the  risk is not apt to be much larger than that predicted by a
linear model" (OSHA 1987).
     Contributing  factors  influencing  cancer  development are  not clearly
understood. Cancer may result  from a multistage  process  occurring  over a
long period of  time, and  presumably initial  and  progressive  stages  of
carcinogenesis may be  modified by  both genetic and environmental  factors
(Strong  1977). Goldstein  (1977)  questions whether benzene  may be an

-------
92   Section 4

Inducer or cocarcinogen  rather  Chan a direct carcinogen, and Cronkite
(1986) suggests that  there  is probably some repair of genetic damage
that leads to neoplastic change.

     Only a small proportion of exposed individuals actually develop
leukemia; therefore it could be interpreted that there is a sensitive
subpopulation, possibly  with some metabolic idiosyncrasy that allows the
formation of reactive metabolites at a specific cellular target (Dean
1985). A familial connection in benzene-related leukemia was established
by Aksoy (1985a,b). Blattner et al. (1976) describe a family involving a
father and four of five  siblings with chronic lymphocytic leukemia. They
conclude that an inherited  defect in a specific class of cells appears
to underlie susceptibility  to leukemia in this family. Although the
authors do not mention benzene  exposure, Goldstein (1977), in a review
of this work, points out that all had been employed in the dry-cleaning
business since the 1940s, a period during which benzene was widely used.
He suggests that the omission of this occupational exposure to benzene
may be evidence of a  "generally lesser degree of physician recognition
of the association of benzene with leukemia."

     Many mechanisms have been  suggested for benzene carcinogenicity.
Leong (1977) has suggested  that, through alterations in lymphocyte
populations, benzene and its metabolites may modify "immune
surveillance" and allow  the development of unusual cellular species that
may lead to the development of  leukemia and other neoplasms in humans.
     Experimental data in animals and studies of human cases of benzene
intoxication indicate a  link between nononcogenic suppression of
cellular growth and the  development of leukemia. Many cases of benzene-
induced leukemia appear  to  have been preceded by aplastic anemia (Toft
et al.  1982). The compensatory  response (regenerative hyperplasia)
observed in the bone marrow, thymus, and spleen of exposed animals may
play a role in the carcinogenic response (Rozen and Snyder 1985, C. A.
Snyder 1987). Harigaya et al. (1981) suggest that benzene may act as a
promoter, rather than an initiator, by forcing proliferation of the
hematopoietic pluripotential stem cell to maintain essential
hematopoiesis and thus hasten the appearance of preleukemic and
leukemogenic clones from stem cells that have been exposed to
leukemogenic initiaCing  agents  prior to benzene exposure.

     One of the favored  mechanisms for benzene toxicity or1-
carcinogenicity is related  to the covalent binding of benzene
metabolites to cellular  macromolecules. In mice administered
radiolabeled benzene for relatively short durations, metabolites have
been found covalently bound to  liver, bone marrow, kidney, spleen,
blood,  and fat (R. Snyder et al. 1978, Gill and Ahmed 1981, Longacre et
al. 1981a).  The label was bound to the nucleic acids of the
hematopoietic cells and  to  nucleic acids and other macromolecules of the
mitochondria (Gill and Ahmed 1981), and, as the levels of covalently
bound and water-soluble  metabolites increased, so did benzene toxicity
(Longacre et al. 1981a).

     Lutz and Schlatter  (1977)  observed covalent binding to DNA in the
livers of rats exposed to benzene vapor. Phenol, hydroquinone, catechol,
benzoquinone, and 1,2,4-trihydroxybenzene form adducts in bone marrow
mitochondria, resulting  in  the  inhibition of the synthesis of

-------
                                                 Toxicologies! Data   93

mitochondrial proteins that are necessary for mitochondria! function
(Kalf et al. 1982, as reported in Andrews and Snyder 1986).

     The ultimate mechanism for benzene-induced hematotoxicity is not
known at this time; it could be one of the mechanisms discussed above
but most likely involves a combination of factors. Identification of the
mechanism would be -facilitated by clear identification of the specific
target cell and intracellular target of benzene and its metabolites
(Andrews and Snyder 1986).
     For additional information on the carcinogenicity of benzene.
summaries and reviews of some of the animal studies can be found in I ARC
1982, Dean 1985, Maltoni 1983, Lee et al. 1983, Van Raalte and Grasso
1982, and NTP 1986. Some summaries and reviews of the human data can be
found in Goldstein 1977, Lee et al. 1983, R. Snyder 1984, Aksoy 1985a,
Aksoy 198Sb, and Kalf et al. 1987.

-------
                                                                      95
               5.  MANUFACTURE. IMPORT, USE,  AND DISPOSAL

5.1  OVERVIEW
     Annual U.S. production of benzene is in excess of 1 billion
gallons, accounting for over 30% of the total produced worldwide.
Benzene is obtained almost entirely from petroleum sources.  It is  used
primarily as a chemical intermediate in the manufacture of various
plastics, synthetic rubbers and fibers, and resins. Such materials are
used in a wide variety of consumer goods, including plastic containers,
radios, toys, sporting goods, furniture, appliances, automobiles,  tires,
adhesives, and textiles. Other uses for benzene and/or its derivatives
include dyes, drugs, pesticides and other agricultural chemicals,
lubricants, solvents, and cleaning products.  Benzene also occurs in
gasoline at concentrations averaging less than 1%, but which can be as
high as 5%. On a volume basis, probably most of* the benzene produced
remains in petroleum fuels such as gasoline.

5.2  PRODUCTION
     More than 90% of the benzene produced in the United States is
derived from petroleum sources. Benzene is obtained from refinery
streams (catalytic reformates), pyrolysis gasoline, and toluene
hydrodealkylation. During catalytic reforming, naphthenes and paraffins
in naphtha are converted to aromatic hydrocarbons, and the benzene is
recovered by solvent extraction (e.g., with sulpholane or tetraethylene
glycol). Pyrolysis gasoline, which is  a by-product generated when
ethylene and propylene are produced by the steam cracking of lower
paraffins or heavier hydrocarbons, is  a mixture of saturated
hydrocarbons, monoolefins, diolefins,  styrenes, and aromatics,  including
a high percentage of benzene  (Hughes and Abshire  1983). In the  toluene
hydrodealkylation process, toluene or  toluene/xylene mixtures are
reacted with hydrogen at temperatures  up to 730°C  and demethylated to
produce benzene  and methane.  In 1981,  catalytic reformates accounted  for
about 50% of the total U.S. benzene production, pyrolysis gasoline 18%,
and toluene hydrodealkylation 24%  (Hughes  and Abshire  1983). Only about
half the benzene available through catalytic reforming  is extracted;  the
rest remains as  a component of gasoline  (IARC 1982, EPA 1985a). The
average concentration of benzene  in gasoline  is <1% (EPA  1985a), but  it
can be as high  as 4  to  5%  (Holmberg and Lundberg  1985).
     Benzene can also be derived  from  coal in the light oil produced
during coke manufacture. This source accounted  for about  7% of  U.S.
production  in 1981  (Hughes and Abshire 1983).
     Estimates  of U.S.  production of benzene  in 1985,  the most  recent
year for  which  data are available,  range from 1.275 billion gallons
 (C&EN  1987b) to 1.363 billion gallons  (CEH 1987).  A U.S.  benzene

-------
96   Section 5

production level of about 1.5 billion gallons has been predicted for
1987 (C&EN 1987a).

5.3  IMPORT
     Annual imports of-benzene into the United States over past years
have generally ranged from 100 to 200 million gallons.  Imports in 1987
were estimated to total 175 million gallons (C&EN 1987a).  Exports were
thought to be less than 10 million gallons.

5.4  USE

     Benzene recovered from petroleum and coal sources  is  used primarily
as an intermediate in the manufacture of other chemicals and end
products. The major uses of benzene are in the production  of
ethylbenzene, cumene, and cyclohexane. Ethylbenzene (57% of benzene
production volume) is an intermediate in the synthesis  of  styrene,  which
is used to make plastics and elastomers. Cumene (20%) is converted to
phenol, phenol derivatives, and acetone, which are used in the
manufacture of phenolic resins, epoxy resins, nylon fibers, and acrylic
resins. Cyclohexane (12%) is used to make nylon fibers  and resins.  Other
industrial chemicals manufactured from benzene include
nitrobenzene/aniline (5%)., linear alkylbenzene sulfonates  (3%),
chlorobenzene (2%), anthraquinone,  benzene hexachloride, benzene
sulfonic acid, biphenyl, hydroquinone, resorcinol, and maleic anhydride
(CEH 1986).

     In the past, benzene was widely used as a solvent, but this use is
now decreasing (IARC 1982). Less than 2% of the amount produced is used
as a solvent in such products as trade and industrial paints, rubber
cements, adhesives, paint removers, artificial leather, and rubber
goods.  Benzene has also been used in the shoe manufacturing industry,
the rotogravure printing industry,  and chemical laboratories (OSHA 1978,
Mara and Lee 1978, both as reported in EPA 1985a). In the  past, consumer
exposure to benzene occurred through  the use of paint strippers,
carburetor cleaners, denatured alcohol, and rubber cement used in tire
patch kits and in arts and crafts supplies (Young et al. 1978).
Recently, Wallace et al. (1987) reported that benzene emissions could be
detected from such products as carpet glue, textured carpet, liquid
detergent, and furniture wax.  For many solvent uses, benzene has been
replaced by other organic solvents; however, it may still occur as a
trace impurity in these products.

5.5  DISPOSAL

     Waste by-products from benzene production processes include acid
and alkali sludges, liquid-solid slurries, and solids  (Saxton and
Narkus-Kramer 1975, Gilbert et al.  1982). Although benzene is included
in the listing of Resource Conservation and Recovery Act (RCRA)
hazardous wastes  (F001-F005, spent solvents; 51FR6537), it is not yet
subject to the treatment standards required for such wastes, and,
consequently, it may be disposed of legally in landfills.  In the past,
landfilling has been a major method of disposing of benzene-containing
industrial wastes. Estimates of benzene levels disposed of in solid
wastes from the petroleum industry are given in Table  5.1.

-------
                    Manufacture,  Import,  Use.  and  Disposal   97
Table 5.1.  Disposal of petroleum industry wastes containing benzene
Disposal
method
Landfilling
Landspreading
Lagoonmg
Incineration

Total
Fraction
disposed
51.1%
8.4%
397%
0.8%


Solid wastes
(kkg)
5.88 X 10*
0.97 X 108
4.57 X 10*
0.09 X 108


Benzene
(kkg)
116
19
91
2
•710

     Source:  Adapted from Gilbert et al. 1982. based on Jacobs
 (1978) by JRB(1980b).

-------
98   Section 5

     The suggested method of disposing of liquid benzene wastes is
incineration (ITII 1975, Sittig 1981). Small spills can be handled by
absorbing the benzene on paper, evaporating in a glass or metal dish in
a hood, and then burning the paper (ITII 1975).
     Benzene-containing wastevater can be processed in standard
wastewater treatment systems using covered hydrocarbon/water separators
(BLackwood et al. 1979).

-------
                                                                      99
                         6.  ENVIRONMENTAL FATE
6.1  OVERVIEW
     Benzene is released Co Che environment by both natural and man-made
sources, although Che anthropogenic emissions are undoubtedly Che most
significant. Annual benzene emissions from man-made sources are in che
vicinity of 236,000 metric tons. Chemical degradation reactions,
primarily che reaction wich che hydroxy radical, limit the atmospheric
residence time of benzene Co only a few days--and possibly only a few
hours--if che concentration of hydroxy radicals is sufficiently high.
Biodegradacion, principally aerobic, is Che mosC importanC environmental
face mechanism for waCer- and soil-associated benzene and, although
probably somewhat slower Chan atmospheric chemical reactions, under the
right condicions can be quiCe efficient.

6.2  RELEASES TO THE ENVIRONMENT
     As che data in Sect. 7.2 indicate, benzene is being released into
the environment. However, since benzene is released both from natural
sources [e.g., crude oil seeps  (Brief et al. 1980), forest fires, and
plant volatiles (Graedel 1978)] and man-made sources, careful
consideration of monitoring data is necessary. Available data suggest
that che benzene levels recorded in rural areas may be che result of
biological sources, while in urban areas the predominant sources are
probably man-made. If Chis is true, Chen seasonal variacions in benzene
release Co rural environmencs would be expecced because of che more
active biological accivicy in peak growing seasons.

     The most significant source for release of benzene Co che
environment is undoubcedly from che combusCion of gasoline. An
indicaCion of Che magnitude of  release can be obtained Chrough
consideration of the firsc entry in Table 6.1. It is likely that a
significant amount of the estimated 40 to 80 thousand metric tons are
released from combustion, but the exact fraction cannot be calculated
from Table 6.1. Table 6.2 presents an estimate of the annual emission of
benzene to water, and Table 6.3 provides a more detailed  look at the
potential sources of benzene released to water. Other sources of
environmental release, although minor compared  to Chose lisced  in
Tables 6.1 and 6.2, include effluents from septic tanks (Viraraghavan
and Hashem 1986), structural fires  (Lowry et al. 1985), off-gassing from
particleboard  (Glass et al. 1986), and exhaled  air of smokers  (Wester et
al. 1986) as well as from cigarette smoke itself (Lauwerys 1979, as
reported in IARC 1982).

-------
100    Section  6
                        Table 6.1.  Annual emissions of benzene to air
                          from various sources in the United States
                         Source                  Emissions (thousand metric tons)
          Component of gasoline"                            40.0-80.0
          Production of other chemicals                      44 4-56.0
          Indirect production of benzene*                     23.0-79.0
          Production of benzene from petroleum                1.8-7.3
          Solvents and  miscellaneous sources                   1.5
          Imports of benzene                              0.013
              "Production, storage, transport, vending, and combustion.
              *Coke ovens, oil spills, nonferrous metals manufacturing, ore mining,
          wood processing, coal mining, and textile industry.
              Source:  Adapted from JRB Associates, Inc.  I980a, as reported in
          IARC  1982.
             Table 6.2.  Annual emissions of benzene to water in the United States
                             Source                    Emissions (metric tons)
             Indirect production of benzene"                     200-11,000
             Solvent and miscellaneous uses                   1450
             Production of chemicals other than benzene        1000
             Production of benzene from petroleum               630
             Imports of benzene                                 13
                 "Coke ovens, oil spills, nonferrous metal manufacture, ore min-
             ing, wood processing, coal mining, and textile manufacture.
                 Source: Adapted from JRB Associates,  Inc. 1980a, as reported
             in IARC 1982.

-------
                                                   Environmental  Face    101
            T»We 6.3. Benzene concentrations in wastewalers
                                            Concentration (ppb)
    Type of wastewater
                           Occurrence
High
                                                  Medium    Low
Timber products
Steam electric
Leather tanning
Iron and steel manufacturing
Petroleum refining
Nonferrous metals
Paint and ink
Printing and publishing
Ore mining
Coal mining
Organic* and plastics
Inorganic chemicals
Textile mills
Plastics and synthetics
Pulp and paper
Rubber processing
Auto and other laundries
Pesticides manufacturing
Photographic industries
Pharmaceuticals
Explosives
Battery manufacturing
Plastics manufactunng
Foundries
Porcelain/enameling
Aluminum
Electronics
Oil and gas extraction
Organic chemicals
Mechanical products
Transportation equipment
Synfuels
Public-owned treatment works
Rum industry
17
21
8
6
43
35
122
22
2
30
58
60
5
17
45.
14
8
17
5
II
17
2
1
2
5
2
17
16
27
4
2
2
24S
6
150025
78819
2274 46
17600
7649 25
29169
403848
83100
20281
757106
303421
5367 79
29044
53765
2645 33
664158
41527
78585
4986
381601
5086 90
56431
142.80
12893
807
25399
281943
9012.57
95752
19515
1482.20
2660
500312
16.83
6448
2847
503
1487
3774
29 17
3592
2505
10191
2184
9494
1953
14.15
1793
2506
9730
964
3785
1626
3002
36230
448 15
14280
6945
374
13072
8843
9126
12.47
625
79631
1795
2973
879
1549
2 12
1 76
524
209
042
1 71
1 37
102
106
1 68
097
503
148
089
263
1 80
190
8 18
169
18 IS
33199
14280
997
273
744
282
1195
078
424
11043
930
0.72
390
    "Number of samples containing benzene
    Source: Unpublished data from EPA survey; see Shackelford and Chne
1983 for discussion of methodology

-------
L02   Section 6

6.3  ENVIRONMENTAL FATE

     A tremendous volume of literature exists concerning the
environmental fate of benzene. This section will cite only a sufficient
number of these references to illustrate certain points;  the reader is
directed to the following for more detailed information:  CHEMFATE (1987,
an on-line database available through Syracuse Research Corporation);
Versar, Inc. (1979); and Korte and Klein (1982).

6.3.1  Transport

     The volatility and solubility of benzene are properties with the
greatest influence on environmental transport of benzene.  On the basis
of solubility data (1,000 mg/L at 25°C) benzene can be said to have a
fairly high solubility in water. It is considered to be a high
volatility chemical with a vapor pressure of 95 mm Hg at 2S°C (Mackay
and Leinonen 1975).

     For benzene released to the air some washout in rainwater is
anticipated. Once benzene in rainwater is deposited in soil or water,
volatilization is expected to return a portion back to the atmosphere,
starting the cycle again. It is conceivable that under the proper
environmental conditions, e.g., no rain and strong winds,  benzene could
be transported several miles; however, because of its reactivity with
the hydroxy radical, global transport of benzene is not likely.
     When released to water, volatilization will result in a substantial
loss to the atmosphere. This potential loss is evident from the Henry's
law constant of 5.5 x 10*3 atm-m^/mol calculated by Mackay and Leinonen
(1975) [Smith et al. (1980) consider chemicals to have high volatility
if this constant is greater than 4.6 x 10'3 atm-m3/mol].  Hackay and
Leinonen (1975) estimated a half-life for benzene of 4.81 h for a
1-m-deep body of water at 25°C. Similarly, Branson (1978) estimated a
half-life of 4.8 h under the same conditions.

     No information was found assessing the potential for benzene
transport to sediments. However, this is not viewed as a significant
transport process because 'the volatility of benzene would favor release
to the air, especially in waters where there is frequent exchange
between surface and bulk water. Branson (1978) notes that the chemicals
that will most likely be associated with sediments are those with low
vapor pressure and low water solubility, characteristics that do not
apply to benzene.

     Benzene released to soil can be transported to the air through
volatilization, to surface water through runoff, and to groundwater as a
result of leaching. For the first two situations to occur, the release
would have Co be at or near the soil surface. If the released benzene is
buried in the earth, then the most likely transport mechanism will be
leaching to groundwater. A useful parameter for investigating the
leachability of a chemical is the soil sorption coefficient (KOc)•
According to Kenaga (1980), compounds with a KOC of <100 are considered
to be moderately to highly mobile. Thus, benzene with a KOC of 60 to 83
(Kenega 1980, Karickhoff 1981) would be considered mobile. Other than
the KOC, other parameters that must be considered to determine if
benzene will reach groundwater include the soil type (e.g., sandy vs

-------
                                                Environmental Face   103

clay),  Che amount of rainfall, the depth of the groundwater,  and the
extent of degradation.

6.3.2  Transformation and Degradation
     Degradation of benzene occurs through both chemical and biological
mechanisms; chemical degradation of benzene is the most important
process for atmospheric benzene and biodegradation most significant for
water- and soil-associated benzene.

6.3.2.1  Chemical degradation
     There is little question that the most significant chemical
degradation process for benzene is its reaction with the atmospheric
hydroxy radical. This can easily be seen by calculating the half-lives
for the oxidation of benzene using the following method:
                             fOH - r-


                      t 4 - 0.693r (Lyman 1982) .

     For OH radicals a half-life for benzene of 5.6 days can be
calculated using a value of 1.3 x 10'12 cm3/molecule•s for kQH (Gaffney
and Levine 1979) and a value of 1.1 x 106 molecules/cm3 for the
concentration of OH radicals (Lyman 1982). When an OH radical value of
1 x 10& molecules/cm3, corresponding to what might be present in a
polluted atmosphere, is used (Lyman 1982), the half-life is shortened to
1.48 h. The same method can be used for other active species such as 03
and 0(3P), but, in contrast, a half-life of 327 years for rural
atmospheres and 104 years for urban atmospheres is calculated for the
reaction of benzene with 03 using a rate constant for 03 of 7 x 10'23
cm3/molecules-s (Pate et al. 1976) and atmospheric concentrations for 03
of 9.6 x 1011 molecules/cm3 (rural) and 3 x 1012 molecules/cm3 (urban)
(Lyman 1982). Similarly, a half-life of 10.9 years for the oxidation of
benzene by 0(3P) radicals can be calculated using 7.2 x 10^
molecules/cm^ as the atmospheric concentration of 0(3P) radicals (NRC
1980b, as reported in EPA 1983) and a value for k  3 . of 0.28 x 10'13
cm3/molecule•s  (Gaffney and Levine 1979).         l  '
     Additional atmospheric reactions include those with nitrogen oxides
and sulfur dioxide, but, similar to reactions with 03 and 0(3P)
radicals, these reactions are secondary in importance to that of benzene
with OH  radicals (Korte and Klein 1982). The reaction of benzene and
nitrogen oxides has been well investigated (e.g., Kopczynski 1964, Levy
1973, Nojima et al. 1975, Korte and Klein 1982), primarily to determine
the role of benzene in photochemical smog formation. The conclusion is
that benzene probably does not play a significant role  in photochemical
smog formation. However, as noted by Nojima et al.  (1975), some of the
products of  the reaction of benzene with nitrogen oxide  (laboratory
investigations) (e.g., nitrobenzene, o- and p-nitrophenol, and 2,4- and
2,6-dinitrophenol) may have potential adverse  effects  to human health.

     Not much  information  is  available  concerning the  chemical
degradation  of benzene  in water.  However, what data are available
suggest  that chemical degradation does  not play an  important role  in  the

-------
104   Section 6

fate of waterborne benzene. Using a rate constant for the reaction of
benzene with OH [(kQH of 0.31 x 1010 L/mol-s (Anbar and Neta 1967)], an
OH radical concentration in water of 1 x 10'17 mol/L (CHEMFATE 1987),
and the equation presented earlier in this section, a half-life of 0.71
year is calculated--much slower than the same reaction in air.
     The direct photolysis of benzene in the atmosphere is not likely
since the upper atmosphere effectively filters out wavelengths of light
<290 run, and, according to Bryce-Smith and Gilbert (1976, as reported in
EPA 1983), benzene does not absorb wavelengths of light >260 run.

6.3.2.2  Biodegradation
     The degradation of benzene by microorganisms has been well
researched, and the conclusion reached is that benzene is biodegradable
(e.g., see Haider et al. 1981, Hopper 1978, Setzkorn and Huddleston
1965, Tabak et al. 1981, Gibson 1977, Higgens et al. 1980, Smith and
Rosazza 1974, Korte and Klein 1982, Unger and Claff 1985).
     The above-mentioned studies document the aerobic degradation of
benzene, and, although far less information is available, benzene
apparently is biodegraded under anaerobic conditions although probably
somewhat slower than aerobically. One study that illustrates this is the
research of Wilson et al. (1986). These investigators found that under
anaerobic conditions in the laboratory, benzene was not significantly
degraded during the first 20 weeks of incubation, but, by 40 weeks of
incubation, benzene concentrations were reduced by 72%. At 120 weeks of
incubation over 99% degradation had taken place. However, Batterman
[1986, as reported in Chea. Abscr. 104(24):212909U], in investigating
the in situ anoxic biological treatment of a hydrocarbon-contaminated
aquifer, reported the complete removal of benzene after only 6 months.
Gibson (1980) presents a proposed pathway for the anaerobic
biodegradation of aromatic compounds.
     As discussed by Gibson (1980) and Hopper (1978), microbial
metabolism of benzene proceeds through the formation of cis-dihydrodiols
and, with further oxidation,  to catechols which are the substrates  for
ring fission. Thus, before going to catechol, benzene biodegrades to
l,2-dihydroxy-1.2-dihydrobenzene (Gibson 1980).

     One important point must be made. The results of laboratory
experiments which characterize most of the above, especially  those  using
large numbers of organisms known to degrade benzene, must be  carefully
applied to field situations.  An example of this is  seen  in the studies
of Haider et al.  (1981). Nocardia  species  and Pseudomonas species,  after
cultivation on benzene, effectively degraded benzene after 7  days (45  to
90%); however, when 100 g  soil with a mixed bacteria population was
mixed with 2 mg benzene, only 47%  of the added  radioactivity  was
recovered as C02 after  10  weeks. Haider et al.  concluded  that specific
organisms which degrade benzene were present  in the soil  in only small
numbers.

-------
                                                                      L05
                     7.  POTENTIAL FOR HUMAN EXPOSURE

 7.L  OVERVIEW

     A  large segment of the U.S.  population  is exposed to benzene. This
 exposure occurs primarily  as  a  result of benzene emitted to the air from
 man-made sources but also  through drinking contaminated water, eating
 certain foods, and  smoking cigarettes. Benzene has been found in at
 least 337 of 1,177  NPL  hazardous  waste sites. Although a large volume of
 benzene is released to  the environment (see  Sect. 6), environmental
 levels  are low due  to efficient environmental removal processes. The
 magnitude of exposure is greatest for those  individuals occupationally
 exposed to benzene;  however,  a  far greater number of individuals are
 exposed as a result of  benzene  released from gasoline filling stations,
 from smoking tobacco products,  and from auto exhaust.

 7.2  LEVELS MONITORED OR ESTIMATED IN THE ENVIRONMENT

     Benzene is ubiquitous in the environment. It has been identified in
 soils,  the aqueous  environment  (freshwater and saltwater), finished
 drinking water, groundwater,  the  air  of both rural and urban
 environments, indoor air,  tobacco smoke, and foods. The following
 subsections provide  relevant  examples of the presence of benzene in each
 of these compartments.

 7.2.1  Air

     As might be expected  from  the use of benzene as a solvent and as a
 component of gasoline,  the presence of benzene in air has been
 extensively documented. Table 7.1 lists a selected number of these
 reported instances.

     These examples  were, selected because they were among the more
 recent  reports of benzene  levels. An  exception is the report by Holzer
 et al.   (1977),  which was chosen because it illustrates the difference in
 levels of benzene for a rural area compared  to an urban environment
within  the sane geographical  area. In the more recent references for
urban air,  average values  were  generally <5  ppb. In contrast, Holzer et
 al. (1977)  reported  an average  value  of 25 ppb for the air of
Tuscaloosa,  Alabama, but it is  not known if  the current level would be
 similar. The highest value in Table 7.1 is 64.6 ppb measured in the air
over Pittsburgh,  Pennsylvania,  during the spring of 1981. The reader is
directed to IARC (1982) and to  EPA (1985a) for measurements made in the
vicinity of point sources  or  associated with automobile use; most, if
not all, of these are pre-1980, and their relevance for current emission
 levels is not known. The highest  value, 9,900 ppb, was measured near a
gasoline bulk-loading facility  and the second highest, 9,400 ppb, near a
similar activity,  the loading and discharging of gasoline from

-------
106
Section 7
                          Table 7.1.  Benzene levels in air samples
         Location
                      Concentration
                      •   (ppb)
      Comments
    References
                                       Ambient air
 San Francisco. California
                    0.8 to 5 2
                    (26  ± 1 3)a
 Stmson Beach. California   0 38 ±  0.39"
 New Jersey, north-
 eastern area
 Houston. Texas


 St. Louis, Missouri


 Denver


 Riverside. California


 Staten Island


 Pittsburgh


 Chicago


 73 km northwest of
 Denver
 Tuscaloosa, Alabama
 Talladega
 National Forest
                    3.9
                            1.4
                            1.3
                    0.84 to 37.7
                    (5.78  ± 5.88)*

                    Oil to 5.82
                    (1.41  ± 1.19)*

                    0.11 to 23.91
                    (4.39  ± 3.94)*

                    0 52 to 10.98
                    (3.95  ± 1.91)*

                    0.082  to 19.034
                    (4.204 ± 4.287)*

                    0.392  to 64.619
                    (5.003 ± 9.818)*

                    0.588  to 8.771
                    (2.561 ± 1.779)*

                    0.02 to 0.85
                    16 to 60
                    (25)'


                    0.2 to 1.3
Results from six
different urban
locations
21 samples;
nonurban area
Industrial area,
241 samples;
Residential
area, 49 samples;
Residential
area, 40 samples;
1978 sampling
period
Spring  1980


Spring  1980


Summer 1980


Summer 1980


Spring  1981


Spring  1981


Spring  1981


Rural area;
sampling between May
1981 and December
1982
Urban area;
sampling in February
1977

Rural area;
sampling in February
1977
Wester et al. 1986
Bozelli and
Kcbbckus 1982
Singh et al. 1982
Singh et al. 1982
Roberts et al. 1985
Holzeretal. 1977

-------
                                            Potential for Human  Exposure
                                                         107
                                Table 7.1  (continued)
       Location
 Concentration
     (ppb)
                                                 Comments
                           Reference
Elizabeth and
Bayonne, New Jersey
Elizabeth and
Bayonne. New Jersey
                                    Ambient air (continued)
2.7 (0.37)'
28.25 maximum
2.9 (0.25)d
13.66 maximum
Night, 81-86
samples
Day. 86-90
samples; Fall 1981,
conversions from
Mg/m3 made
                                      Indoor air
9.6 ± 1 55,
158.3 maximum

8.38 ± 0.43.
83.82 maximum
Night, 346 to
348 samples

Day, 339 to 341
samples;
Fall 1981 sampling
period; /ig/m3
converted to ppb
                                                                   Wallace et al. 1985
                                                                   Wallace et al. 1985
   "Average ± standard deviation.
   * Arithmetic average ± standard deviation.
   'Average.
   ''Weighted arithmetic mean (standard error).

-------
 108    Section 7

 road tankers.  Most of the atmospheric benzene  levels documented  In  the
 IARC and EPA documents are,  however, much  less  (e.g., 9-  to  19-ppb
 average concentrations recorded near a cumene manufacturing  plant in
 Philadelphia and an average  of 0.9  ppb measured near the  American
 Cyanamid Company in Linden,  New Jersey). Table 7.1 also lists values for
 benzene in the air of residences in the Elizabeth and Bayonne, New
 Jersey,  areas (see Wallace et  al. 1985). The interesting  aspect  of  this
 study was the observation that the  benzene levels in the  indoor  air were
 greater than those recorded  in the  air outside the residences. In a
 follow-up report,  Wallace and  Pellizzari (1986) present a comparison of
 benzene levels in indoor  air from homes occupied by smokers vs homes
 without smokers.  In the fall and winter, homes with smokers  (N - 345)
 had  average  benzene concentrations  of 52 ppb compared to  30 ppb  in homes
 without smokers (N - 164). In  spring and summer, the levels were
 comparable (about 15 ppb)  probably  due to houses being more open. This
 points to the  possible significance of passive smoking as a source of
 benzene exposure.

 7.2.2   Water

     Representative examples of benzene concentrations that have been
 measured in  rainwater,  surface  water,  sea water, drinking water, and
 groundwater  are shown in  Table  7.2. Measurable levels range from
 0.005  ppb in the  Gulf of  Mexico to  330 ppb in contaminated well water of
 New York,  New  Jersey,  and Connecticut.-  Not shown in Table 7.2 is an
 atypically high concentration  of benzene of 24,000 ppb which was found
 in subsurface  water samples  taken near extensive gas and oil deposits
 (Ochsner et  al.  1979,  as  reported in IARC 1982).

 7.2.3   Soil

     The only  measured values  of benzene found for soil was in a study
 by Fentiman  et al.  (1979), who  recorded levels ranging from <2 to 191 ppb
 in the vicinity of five industrial  facilities using or producing benzene.

 7.2.4  Other

     Benzene has been identified in several types of foods [e.g.,
 Jamaican rum  (120  ppb), irradiated  beef (19 ppb),  and heat-treated
 canned beef  (2  ppb)]  (NCI  1977,  as  reported in EPA 1985a). EPA (1985a),
 after reviewing a  number  of  sources, provides the following
 nonquantitative list  of benzene-containing foods:  haddock fillet, dry
 red beans, blue cheese, cheddar  cheese,  cayenne pepper,  pineapple,
 roasted  filberts,  potatoes (cooked  peels),  cooked chicken, hothouse
 tomatoes,  strawberries, black currants,  roasted peanuts,  soybean milk,
 and codfish.

     Lauwerys  (1979,  as reported in IARC 1982) found that cigarette
 smoke contained 0.047  to  0.064  ppb  of benzene. Similarly,  Wester et al.
 (1986) observed that  the breath  of  smokers (residents of the San
 Francisco  area) contained higher levels of benzene (6.8 ± 3 ppb) than
 the breath of  nonsmokers  (2.5 ±  0.8 ppb).  More dramatic differences in
 the breath of  smokers  compared with that of nonsmokers was seen by
Wallace  and Pellizzari  (1986), who  report average benzene breath levels
of 51 ppb  for  smokers  (N - 198)  and 8 ppb for nonsmokers  (V - 322).

-------
                                                      Potential  for Human Exposure
                                                             109
                                 Table 7.2.  Benzene levels in water samples
Source type Location
Rainwater United Kingdom.
Surface water United States

Concentration
(ppb) Comments
872
NQJ River and lake water
1 to 7 Heavily industrialized
river basins. 40 of 240
sites positive, sampling
from August I97S through
September 1976
References
Colenutt and
Thorburn 1980
Shackelford and
Keith 1976
Ewmg et al 1977.
as reported in
CHEMFATE 1987
Sea water
                Lake Erie
                United States
                Lake Michigan
                Lake Zurich,
                Switzerland

                United Kingdom
                Gulf of Mexico
Drinking water   United States
                Ontario
                                       Oto I
I to 13
(4 avg )


I to 7
0028


6 5 to 8 9


0005 toOOlS


0005 to 0175


NQ

0 I to 0 3

 < 0 2 (avg )

 SO


04
                One of two sites
                positive, sampling during
                I97S and 1976
13 sampling locations
upstream and downstream
near industrial outfall

Five of nine sites
positive, sampling during
I97S and 1976

Surface water
Lake water


Unpolluted waters.
sampling during 1977

Polluted waters.
sampling dunng 1977

24 documented occurrences
 7 of 113 sites positive

 Presumably an average
 of 700 1975 samples

 Absorption/desorption of
 benzene from anthracite
 coal screenings used as
 Tilter medium
Konasewich et al
1978. as reported
in CHEMFATE 1987
and HSDB 1987

Fentiman et al
1979


Fentiman et al
1979


Grob and Grab 1974


Colenutt and
Thorburn  1980

Sauer  1981


Sauer  1981


Shackelford and
 Keith  1976

 EPA I980b

 Brass et al. 1977

 Kraybill 1977


 Smilheeial  1978
Groundwater


Groundwater
New York, New Jersey.
Connecticut
United States

United States
30-330
> 100
NQ
NQ to 100
Contaminated
well water
Well water from
Jacksonville. Florida
8 5% occurrence in a
federal survey
Found in leachate from some
residential wells adjacent
to a landfill; deep drinking
well had highest concentration
Burmaster 1982. as
reported in
CHEMFATE 1987
EPA I980b
Dyksen and Hess
1982
Stuart et al.
1985
    *NQ - not quantified.

-------
110   Section 7

7.3  OCCUPATIONAL EXPOSURES
     Without question, individuals employed in industries which use or
make benzene are at the greatest risk of encountering potential adverse
health effects from benzene exposure. It has been estimated that
2,000,000 to 3,000,000 persons in the work force are potentially exposed
to benzene (NIOSH 1974, NTP 1985). In a 1985 Federal Register notice
(OSHA 1985), OSHA presents a table (see Table 7.3) illustrating some of
the types of industries and associated levels of benzene exposure and
the number of individuals exposed. The number of estimated workers
exposed is less than that given by NIOSH but may be partially explained
in that not all workers and associated job categories are represented. A
significant number of workers (17,336) are potentially exposed to 8-h
TWA concentrations of benzene ranging from 5.1 to 10.0 ppm. Using
10 ppm, a daily exposure level on a milligram per kilogram basis can be
calculated. Assuming a 70-kg adult and a 30% systemic absorption rate,
and using 10 m3 as the daily intake of air, a daily intake of 1.4 mgAg
is determined. Dermal absorption was not considered in the above
calculation. Although various studies, such as those of Franz (1984) and
Susten et al. (1985), indicate that dermal absorption of liquid benzene
can occur (see Sect 4.2.2.3 for a detailed discussion of dermal
absorption), no data were found for the dermal absorption of benzene
vapor; inhalation is considered to be the most significant route of
exposure to the chemical.

7.4  POPULATIONS AT HIGH RISK
     Other than individuals who are occupationally exposed, discussed  in
the preceding section, special risk populations include those living
near certain chemical manufacturing sites, cigarette smokers, and
individuals living adjacent to landfills. For persons living near
landfills, the extent and media-associated exposure will depend on
whether their drinking water comes from groundwater and whether the
discarded benzene is buried sufficiently deep to eliminate the
possibility of volatilization to  the air. For these three  special risk
populations, some estimates of daily intake can be generated. From  the
data presented in Sect.  7.2.2., it is not unreasonable to  assume that  an
individual could be exposed to 100 ppb in drinking water  (well water).
Thus,  for the average daily water consumption of  2 L, an  individual
would  consume 200 Mg/day benzene  or, for a  70-kg  adult, 2.86 /ig/kg/day.
     With respect to  cigarette smoking, EPA (1985a), using the data of
Newsome et  al. (1965, as reported in EPA 1985a), which showed  that  a
40-mL  draw  of cigarette  smoke contained 6.1 /ig  of benzene, and assuming
15 draws per cigarette,  20 cigarettes smoked  per  day, and a  daily  air
intake of 20 m3, calculated that  the equivalent annual average
atmospheric  exposure  would be 92  pg/m3  (28  ppb).  On  a weight/weight
basis, a 70-kg individual who smokes 20 cigarettes a day  would take in
7.8 /ig/kg/day  (assuming  a systemic benzene  absorption rate of  30%).
     Mara and Lee  (1978, as reported in EPA 1985a) have  estimated that
as many as  200,000 people may be  exposed  to annual benzene average
concentrations of 4.1 to 10.0 ppb and another 80,000 to  annual average
concentrations of >10 ppb as  a  result of  living in the vicinity of

-------
                                                    Potential  for  Human  Exposure     111
                 Table 7.3. Number of employees exposed to beueae (by exposure levels
                                     sad by industry divisions)"
8-hour TWA benzene concentrations (ppm)
Industry sector
Petrochemical producers
Petroleum refineries
Coke and coal chemical
Tire manufacturers
Bulk terminals
Bulk plants
Transportation via tank truck
Total of all sectors
000-010
5.460
36.510
—
34.645
14.556
26.845
32.558
150.574
011-050
4.064
14.751
3.550
24.375
8,260
15.234
10.996
81.230
051-10
1.224
2,600
422
4,095
1,335
2,461*
2.523
14.660
1 1-50
1.212
2.148
436
1,820
932
1.7I8C
1.380
9.646
5 1-100
159
283
71
0
1,988
14,787
48
17.336
100 +
122
226
18
0
25
46C
95
533
Total*
12.242
56.517
4,499
65.000
27,095
61.093
47.600
274,047
    "This table summarizes the worker groups for which OSHA has good exposure data. However, due to the
ubiquitous nature of benzene (e.g., it is naturally occurring constituent of crude oil. natural gas. and coal),
OSHA was not able to obtain exposure data for some worker groups exposed to benzene such as od and gas
well drillers.
     Figures do not total in some cases due to rounding.
    rNumber in original table was in error, number presently in table obtained as a result of personal com-
munication with OSHA)

    Source Adapted from USOSHA 1985

-------
112   Section 7
chemical manufacturing plants. Exposure levels of 4.1 to 10.0 ppb would
yield a body burden of 1.13 to 2.76 ^g/kg/day for a 70-kg man (assuming
a 30% systemic absorption factor).

-------
                                                                     113
                         8.  ANALYTICAL METHODS

     The analytical methods commonly used to identify and measure
benzene levels in various environmental and biological media are given
in Tables 8.1, 8.2, and 8.3. Recommended methods are briefly reviewed in
the following sections.

8.1  ENVIRONMENTAL MEDIA

8.1.1  Air

8.1.1.1  Sample collection and preparation
     Air samples for benzene analysis are usually preconcentrated by
passing the sample through an adsorbent or trap. Commonly used
adsorbents are Tenax resin, silica gel, activated carbon, and
carbonaceous polymeric compounds. Benzene can also be preconcentrated by
cryogenic trapping (Table 8.1).
     Benzene, as a volatile organic, presents difficulties in collection
and handling methods. These problems are discussed in the following
additional references: Withey and Martin (1974), Baslet (1974), Angerer
et al. (1973).

8.1.1.2  Methods
     Gas chromatography (GC) coupled with flame ionization detection
(FID), photoionization detection (PID), or mass spectrometry (MS) is the
most commonly used method for analyzing for benzene in air (Table 8.1).
Of these methods, GC/PID and GC/FID provide a greater degree of
sensitivity than GC/MS, but the latter procedure is more reliable in
identifying benzene in samples containing multiple components having
similar GC elution characteristics. Various spectrophotometric methods
have also been used for determining benzene in  air samples. These are
generally not as sensitive as the GC methods. EPA has sponsored  the
development of an atomic line molecular spectrometer  (ALMS) to monitor
levels of benzene and other organics in ambient air, particularly near
waste disposal and industrial production sites  (Hadeishi et al.  1985).
This method has a detection limit of 250 ppb  (by volume) at 184.9 nm.
     For determining atmospheric levels of benzene in the workplace,
several different analytical methods are acceptable to OSHA (OSHA 1985),
however, the recommended procedure  involves the collection of  the sample
vapors on charcoal adsorption tubes followed  by GC/MS analysis  (NIOSH
Method S-311). Other acceptable methods include portable direct  reading
instruments, real-time continuous monitoring  systems, and passive
dosimeters (OSHA 1985). The latter  methods generally have a sensitivity
in the part-per-million range.

-------
114   Section 8

Sample
matrix
A -bient and/or
oc.upational
atmospheres






































Sample
preparation
Tenax trap. He
desorption

Adsorbent trap.
thermal desorption
Silica gel trap, water
desorption, headspace
analysis
NA

Tenax trap, thermal
desorption, cryogenic trap
Charcoal trap, CSj
desorption
Tenax GC trap.
thermal desorption.
cryogenic focusing
Cryogenic trap
with liquid 02 or Ar,
thermal desorption
Cryogenic trap.
thermal desorption
Charcoal trap.
CSj desorption
Direct injection
Cryogenic trao.
thermal desorction
Cryogenic or Tenax GC
trap, thermal desorption
Cryogenic trap.
thermal desorption
NA
Silica gel trap
Direct analysis
Direct analysis
NA
NA
Silica gel trap



Analytical
method"
GC


GC/MS

GC/MS


GC/FID

GC/FID

GC/FID

GC/FID/MS


GC/FID


GC/FID

GC/FID

GC/PID
5C/PID

GC/PID

GC/PID

Photo.
Spectra
UV Spectra.
UV Spectra.
Spectra.
IR Spectra
Indicator tube:
UNICO tube
AUER tube
DRAGER tube
Limit of
detection
>0 1 ppb


NA

0 1 ppb


<2 5 ppm

0 03 ppb

009 ppb

0003 ppb


<0.06 ppb


NA

<2.5 ppm

0.25 ppb
0.5 ppb

Spot

Ippt

210 ppm
<3S ppm
10 ppm
250 ppb
25 ppb
0 15 ppm
1 5 ppm
10 ppm
5 ppm
5 ppm
Accuracy
NA


NA

NA


+08%
(at 25 ppm)
+ 44%

NA

±60%'


±86%'
(at 1.4 ppb)

±8%rf

+08%
(at 25 ppm)
±51%'
NA

±3.6%rf

NA

NA
NA
NA
NA
NA
NA
±15%
NA
NA
NA
References
Fentunan et al. 1979


Jonsson and Berg 1980*

Gruenke et al. 1986


NIOSH 1977

Martin et al. 1980

Baxter et al. 1980'

Roberts et al. 1984


Singh et al. 1985


Holden et al. 1985

NIOSH 1974, I977b

Clark et al. 1984
Kowalski et aL 1985

Reineke and Baechmann 1985

Nutmagul and Cronn 1985

Du Pont'
Maffett et aL 1956C
Berkley et al. 1984/
Hadeishi et al. 1985
Hager 1973'
Henan 1973*
Koljkowsky 1969'
Matbesoo Scientific*
Auer*
Drager*

-------
                                                                     Analytical  Methods     115
                                             TaMe8.1  (continued)
Sample
matrix
Near spill sites
Landfills/waste sites



Industrial emissions
Sample
preparation
Direct analysis
Carbon trap, carbon
disulfide desorption
Tenax GC trap.
thermal desorption
NA
Analytical
method'
Elect rochem
GC/FID

GC/FID/ECD/MS

GC/FID
Limit of
detection
NA
NA

001 ppb

0 I ppm
Accuracy
NA
NA

NA

NA
References
Stetter et al 1986
Colenutt and Davies 1980C

Harkovetal 1985

Knoll etal 1978
"<-.(- /M<; =» oac rhrnmatnoranhv /mass SOeclrometrv
 GC/FID - gas chromatography/flame lomzation detection
 GC/PID - gas chromatography/photoionization detection
 Photo ™ photometry
 Spectro. - spectrometry
 UV Spectro  — ultraviolet atomic line molecular spectroscopy
 IR Spectro  - infrared spectroscopy
*NA •» data not available.
cAs reported in I ARC 1982.
''Relative standard deviation.
'As reported in Verschueren 1983.
•^As reported in Hadeishi et al  1985.

-------
116     Section  8
                     Table 8.2. Analytical methods for neaswiog benzene k?eb in water and soil
Sample
matrix
Surface water
and/or
effluents


Sample
preparation
N2 purge, Tenax GC trap."
thermal desorption
He purge, polymer trap,
thermal desorption
Inert gas purge, adsorbent
Analytical
method0
GC

GC/MS
GC/MS
Limit of
detection
01 ppb

0 1 ppb
1 ppt
-0 03 ppb
NA
(at 1 ppb)
-1- 11%
(at O.I Mg/L)
NA
NA
NA
NA
NA
NA
-48%
(at 0 1 Mg/L)
NA
NA

Hammers and Bosnian 1986
Bradley and Frenzel 1970
Brass et al. 1977
Stuart et al 1984
Schultze and Kjeldsen 1985
Sawhney and Kozloske 1984
Fentiman et al 1979
Hammers and Bosnian 1986
Ferrano et al 1985
Harland et al 1985
  JGC — gas chromatography.
   GC/MS ~ gas chromatography/mass spectrometry
   GC/FID • gas chromatography/flame wmzation detection
   GC/PID — gas chromatography/photoiomzation detection
   LRS — Laser-Raman spectroacopy.
  *NA - data not available.
  cAs reported in IARC 1981
   Relative standard deviation.
  'Representative measurement and not detection limit.

-------
                                                                   Analytical  Methods     117
           Table 8J. Analytical methods for measuring benzene levels in biological samples and food
Sample
matrix
Blood









Unne

Breath






Tissues

Human milk

Foods
Fruits and
vegetables

Shellfish


"GC-
chromatoKrai
Sample
preparation
Headspace analysis
Headspace collection
cryogenic trap, thermal
desorption
N2 purge, Tenax GC-"
silica gel trap
He purge, Tenax GC trap
Dissolve in pentane
Mix with sodium
citrate solution
NA

Adsorbent gels

Cryogenic trap.
Tenax GC trap.
thermal desorption
Tenax GC trap.
Headspace analysis
Treat with chlorobenzene.
ethanol, and water
He purge. Tenax GC trap.
thermal desorption
NA
Mix with water and
methanol, puree.
azeotropic distillation
Tissue homogenized.
Nj purge, Tenax GC-sdica
gel trap, thermal desorption
gas chromatograpby. GC/MS -
jay/flame lomzation detection.
Analytical
method"
GC/MS
GC


GC/MS

GC/FID
GC/FID
GC/FID

GLC

GC

GC
GC/MS

GC/MS
GC/MS
GC/MS

GC/MS

GC/MS
GC/FID


GC/MS


Limits of
detection
2 ng/mL
I6ppb


OS ppb

NA
08-48 pg/L
20 Mg/L

1 pg/mL.
(phenol)
0 14 ppm

<1 ppb
r-Jppt

35ppt
01 ppb
NA

NA

<10 ppt/
NA


NA



Accuracy
NA*
NA


S<*CVC
(at 2 ppb)
-2%d
NA
NA

NA

NA

NA
NA

±10-30%
NA
NA

NA

NA
-16%
(at 1 Mg/g)

NA



References
Gruenke et al 1986
Wallace 1986


Antome et al 1986

Michael et al 1980
Ghimenti et al. 1978'
Pinigma and Mal'tseva 1978'

LauwenjS 1979. as reported
in van Sittert and deJong 198S
Sherwood and Carter 1970. as
reported in I ARC 1974
Conkleetal 1975
Umanaetal 1985

Wallace 1986
Gruenke et al 1986
Nagataetal 1978'

Michael et al. 1980

NCI 1977
Kozioski I98S


Ferranoet al. 198S


gas chromatography/mass spectrometry, GC/FID — gas




°NA - data not available.
CCV - coefficient of variation
rf2 Mg chlorobenzene spike.
'As reported in IARC 1982.
/Estimated from tabulated data given.
'As reported in Verschueren 1983.

-------
118   Section 8

8.1.2  Water

8.1.2.1  Sample collection and preparation

     Benzene is usually isolated from aqueous media by the purge and
trap method. An inert gas such as nitrogen is used to purge the sample.
The benzene is then trapped on an adsorbent substance, such as Tenax GC
or activated charcoal, and then thermally desorbed (Table 8.2). Liquid-
liquid extraction procedures using solvents in which benzene is more
readily soluble have also been used. The solvent is removed by selective
evaporation. Additional information regarding sample collection and
preparation can be found in a manual recently published by the Office of
Drinking Water, EPA, Volatile Organic Chemicals: Methods and Monitoring
Document (EPA 1987b).

8.1.2.2  Methods

     GC alone, or coupled with FID, FID, or MS, is commonly used to
analyze for benzene in aqueous media (Table 8.2). Sensitivity of these
methods is generally <1 ppb. For the analysis of drinking water, EPA
recommends purge and trap GC (EPA Method 503.1), purge and trap GC/MS
(EPA Method 524.1), purge and trap capillary column GC/MS (EPA Method
524.2), and purge and trap GC with photoionization and electrolytic
conductors in series (EPA Method 502.2) (EPA 1987b). For the analysis of
wastewaters, EPA recommends GC (EPA Method 602) or GC/MS (EPA Method
624).

8.1.3  Soil

8.1.3.1  Sample collection and preparation

     Benzene can be isolated from soil samples by the purge and trap
method, continuous liquid extraction, or Soxhlet extraction.

8.1.3.2  Methods

     Soil samples have been analyzed for benzene using GC and GC/FID
(Table 8.2). A detection limit as low as 1 ppt has been reported for one
method (Hammers and Bosnian 1986).

8.1.4  Food

8.1.4.1  Sample collection and preparation

     Sample collection and preparation for the analysis of benzene  in
foods  includes the purge and trap method, headspace gas analysis, and
azeotropic distillation of samples pureed with water and methanol
(Table 8.3).

8.1.4.2  Methods

     Chromatographic procedures, particularly GC/FID and GC/MS, have
been used to analyze for benzene in  foodstuffs  (Table 8.3). Little
quantitative data are available, but limits of  detection can be <10 ppb.

-------
                                                Analytical Methods   119

8.2  BIONEDICAL SAMPLES

     Analytical methods have been developed to measure benzene levels in
exhaled breath, in blood, and in various body tissues. Urinary analysis
for phenol, a benzene metabolite, has also been used as a measure of
benzene exposure.

8.2.1  Fluids/Ezudates

8.2.1.1  Sample preparation
     Benzene can be extracted from blood samples by several methods
(Table 8.3) including: (1) the purge and trap method using an inert gas
such as nitrogen or helium and an adsorbent such as Tenax GC (Antoine et
al. 1986), (2) cryogenic trapping of headspace gas (Wallace et al.
1986), (3) dissolving in pentane (Ghimenti et al. 1978), (4) mixing with
a sodium citrate solution (Pinigina and Mal'tseva 1978, as reported in
IARC 1982), or (5) mixing with toluene (Snyder CA et al. 1975; Jirka and
Bourne 1982).
     Collection and preparation of breath samples for benzene analysis
usually involves concentrating the sample with adsorbents such as Tenax
GC followed by thermal desorption. Cryogenic traps have also been used.
     For phenol determinations, urine samples are subjected to acid
hydrolysis and extraction.  NIOSH (1984) recommends hydrolyzing the
sample with either concentrated hydrochloric acid or 70% perchloric acid
and extraction with diethyl ether.  Roush and Ott (1977) utilized a
modification of this method in which the hydrolyzed urine was saturated
with sodium chloride before extraction with isopropyl ether.  The
addition of sodium chloride increased the extraction efficiency of the
ether.

8.2.1.2  Methods
     GC coupled with FID or MS is used to analyze for the presence of
benzene in blood (Table 8.3). Limits of detection are <1 ppb. It  should
be noted that current technology limits the interpretation  and the
validity of values for blood levels of benzene generated in various
monitoring programs and their laboratories.
     GC and GC/MS are also used  to measure benzene levels in  exhaled
breath. Limits of detection are  in the part-per-trillion* range.

     An indirect method for evaluating benzene exposure  involves
measuring urinary levels of the  benzene metabolite phenol with gas
liquid chromatography (GLC) or colorimetry. NIOSH  (1984) recommends  the
use of gas chromatography in combination with a  flame  ionization
detector. The estimated limit of detection for  this method  is 0.5 Mg/»L
urine.
     One of the main problems  involved  in measuring urinary phenol
levels to monitor benzene exposure  is the variance  in individual  base
line phenol levels. Roush and Ott  (1977) detected  levels of phenol
exceeding  75 mg/L in  the urine of  individuals who had no known exposure
or whose TWA benzene  exposure was  less  than  5 ppm  (75 mg/L was proposed
by NIOSH,  in  1974, to be an  indication  of unacceptable benzene

-------
120   Section 8

exposure). As mentioned previously, urinary phenol excretions can be
increased by exogenous nonbenzene sources such as dietary protein (Folin
and Denis 1914); medicines that contain phenylsalicylate (Pepto-Bismol®
and Chloraseptic* lozenges) (Kociba et al. 1976); asprin (Fishbeck et
al. 1975); and calamine lotion and phenol-camphor-liquid petrolatum
preparations (Ruedemann and Deichmann 1953).  See Sect.  2.2.2 for further
discussion of biological monitoring of benzene exposure.

8.2.2  Tissues

8.2.2.1  Sample preparation
     Benzene has been extracted from various tissue samples by
dissolving in a mixture of chlorobenzene, ethanol, and water at 60°C
(Nagata et al. 1978, as reported in IARC 1982).

8.2.2.2  Methods
     GC coupled with MS has been used to analyze for the presence of
benzene in body tissues (Table 8.3). Little information is available
concerning sensitivity and accuracy of the method.

-------
                                                                     121
                   9.  REGULATORY AND ADVISORY STATUS

     Regulatory standards and advisory guidelines for benzene are
summarized in Table 9.1 and briefly reviewed in the following sections.

9.1  INTERNATIONAL
     The World Health Organization has recommended a drinking water
guideline for benzene of 0.01 mg/L, a level corresponding to an excess
lifetime cancer risk of 10'5 (WHO 1984).

9.2  NATIONAL

9.2.1  Regulations

9.2.1.1  Media-specific
     Air.  In 1971, OSHA promulgated an occupational exposure standard
for benzene which included a 10-ppm permissible exposure limit (PEL) for
an 8-h TWA concentration, a 25-ppm ceiling limit, and a 50-ppm maximum
(10-min) ceiling limit (OSHA 1971). In 1987, this standard was revised
to 1 ppm for an 8-h TWA concentration and 5 ppm for a short-term
exposure limit (STEL) (OSHA 1987). In the same ruling, OSHA also set 0  5
ppm as the action level for benzene (OSHA 1987).
     Water.  In 1985, the EPA Office of Drinking Water proposed setting
the Maximum Contaminant Level (MCL) for benzene in drinking water at
0.005 mg/L (EPA 1985d). This standard was promulgated in 1987 (EPA
1987a).
     Food.  No information was found indicating whether benzene levels
in foodstuffs are limited by the U.S. Food and Drug Administration
(FDA). Under the Federal Insecticide, Fungicide,  and Rodenticide Act
(FIFRA), benzene is exempt from food tolerance restrictions provided it
is used in accordance with good agricultural practice and only applied
to growing crops (EPA 1985e).

9.2.1.2  Hazard ranking
     Benzene is a priority toxic pollutant, and it has been classified
by EPA as a hazardous substance (Clean Water Act), a hazardous air
pollutant (Section 112 of the Clean Air Act), and a hazardous waste
constituent (RCRA).
     Based on weight-of-evidence of carcinogenicity and carcinogenic
potency, EPA has assigned benzene  a "medium" hazard ranking  (EPA 1986)
However, a hazard ranking is only  for a specific exposure scenario
(e.g., accidental spills) and is not applicable to chronic  low-level
exposures.

-------
122     Section  9
                           Table 9.1.  Regulations and advisory guidance for benzene
                   Concentration
       Medium
 mg/L
                             ppm
Risk
level
            Exposure
            condition
                             Application
                                                                                      References
       Water
001
  InteraatioMl advisory goidanet

                         Drinking water
10  5     Lifetime
                                            National reflation
                                                                                   WHO 1984
Air

Water
1
5
0005
8-h TWA"
STEL*
MCI/
Occupational
Occupational
Dnnkiag water
OSHA 1987
OSHA 1987
EPA I987a
National advisory guidance
Air



Water










Water




01
05
10
10
0
0
0000066
000066
000068
00066
00068
0.068
0.334
12.5

0001




10-hTWA
Action level
15-mia CLrf
8-h TWA
Lifetime
MCLG*
10~* Lifetime
10 Lifetime
10 6 Lifetime
I0~{ Lifetime
10 5 Lifetime
10 4 Lifetime
10-day HA7
7-day SNARL'
Scale refBtatioM
Lifetime

Otter


Occupational
Occupational
Occupational
Occupational
Ambient water
Drinking water
Ambient water
Ambient water
Drinking water
Ambient water
Drinking water
Drinking water
Drinking water
Drinking water

Drinking water
Groundwater

Listed at a
hazardous substance
NIOSH 1986
OSHA 1987
NIOSH 1986
ACGIH 1986
EPA 1980a,b
EPA I984c
EPA !980b
EPA 1980b
EPA I985a
EPA 1980b
EPA I985a
EPA I985a
EPA 1985a
NRC 1980a

FPC/API 1986
(Flonda)

Hazardline 1986
(California)
          'Time-weighted average.
           Soon-term exposure limh.
          c Maximum contaminant leveL
           Celling value.
          'Maximum contaminant level goal
          ' Health advisory.
          'Suggested no-adverse-response level

-------
                                    Regulatory and Advisory Status   123

9.2.L.3  Emission and effluent regulations

     Because benzene is- considered a toxic waste, federal hazardous
waste management procedures must be complied with for any industrial
process involving the generation, transport, treatment,  storage,  and
disposal of the chemical. According to the Comprehensive Environmental
Response, Compensation and Liability Act (CERCLA) of 1980, notification
must be made to the National Response Center when benzene releases into
waterways reach or exceed 1,000 Ib. EPA has proposed that this minimal
reportable quantity (RQ) be decreased to 10 Ib. Benzene is also listed
as a toxic chemical subject to annual reporting of environmental
releases under Section 313 of the Emergency Planning and Community Right
to Know Act of 1986.
     Although benzene has been designated as a RCRA hazardous waste and
is included in the F001-F005 listing of spent solvents (51FR6537), it is
not subject to the recently promulgated treatment standards for such
wastes (51FR40572). It is expected that treatment standards for benzene
will be promulgated by August 8, 1988.
     Priority pollutant effluent limitations [new source performance
standards (NSPS) with best available technology  (BAT)] set the maximum
daily discharge rate for benzene at 0.057 mg/L per day, with a monthly
average not exceeding 0.021 mg/L (EPA 1985b).
     Federal regulations limit daily disposal of benzene in effluents
from coke-production facilities to either 0.0000215, 0.0000250. or
0.0000355 kg/kkg, depending on the type of coke plant. For new coke
plants, daily discharges are limited to either 0.0000319 or 0.0000355
kg/kkg, depending on the type of plant (CFR Title 40, Parts 420.13 and
420.14)
     Benzene is regulated under the Clean Water Act Effluent Guidelines
as stated in 40 CFR Parts 400-475. For each point source category,
benzene is either regulated as part of a group of chemicals controlled
as Total Toxic Organics  (TTO), has a regulatory  limitation provided for
by the particular 40 CFR Part Number  (RL), or has a zero discharge
limitation for the particular 40 CFR Part Number (ZDL). The point source
categories with their effluent limitation designation and specific 40
CFR part numbers, as summarized by the EPA  (1988a), are as follows:
electroplating (TTO, part 413); organic chemicals (RL, part 414); iron
and steel manufacturing  (RL, part 420); steam  electric  (RL, part  423);
asbestos (ZDL, part 427); timber products processing  (ZDL, part 429);
metal  finishing  (TTO, part 433); paving and roofing  (ZDL, part 443);
paint  formulating  (ZDL,  part 446); ink formulating  (ZDL,  part  447); gum
and wood (ZDL, part 454); carbon black (ZDL, part 458); metal  molding
and casting  (TTO, part 464);  and copper forming  (TTO, part 468).  The
effluent guidelines are  examined in  detail  in  40 CFR under the
appropriate part number.
     EPA proposed National Emission  Standards  for Hazardous  Air
Pollutants  (NESHAPS) for five potential benzene  emission sources--maleic
anhydride plants, ethylbenzene/styrene plants, benzene  storage vessels,
benzene  fugitive emission sources, and coke by-product plants. The
proposed NESHAPS for the first  three categories  were withdrawn by EPA in
1984  (49FR23558).  In 1981,  the  EPA proposed a  national standard  for

-------
124   Section 9

fugitive emissions of benzene which would prohibit detectable emissions
from processing equipment that contains materials with benzene
concentrations of 10% or more by weight (EPA 1981, as reported in IARC
1982). NESHAPS was promulgated in 1984 and is set at 10 g/day, not to
exceed an ambient air level of 0.01 /ig/m3. Final action on the proposed
NESHAPS for coke by-product recovery plants is expected in 1987.

9.2.1.4  Consumer products regulations
     In 1978 the U.S. Consumer Product Safety Commission (CPSC) proposed
banning all commercial products (excluding gasoline and laboratory
reagents) containing 0.1% or more benzene (by volume).  However, in 1981
the CPSC withdrew its proposal after determining that the current levels
of benzene did not pose a significant risk to consumers (CPSC 1981).

9.2.2  Advisory Guidance

9.2.2.1  Media-specific
     Air.  NIOSH has recommended that the occupational exposure standard
for benzene be revised to a 10-h TWA of 0.1 ppm (0.32 mg/m3) with a
15-min ceiling value of 1 ppm (3.2 mg/m3) (NIOSH 1986).
     The threshold limit value (TLV) for benzene recognized by the ACGIH
is 10 ppm for an 8-h TWA (ACGIH 1986). A notice has also been given by
ACGIH that the current 25-ppm STEL for benzene will be deleted from its
list of recommended exposure standards.
     Water.  EPA's Office of Water Regulations and Standards is
responsible for establishing water quality criteria for water
pollutants. Water quality criteria are qualitative or quantitative
estimates of the concentration of a substance which, when not exceeded,
will ensure a water quality sufficient to protect a specified end use.
Such criteria are nonenforceable recommendations which do not take into
consideration economic or technical feasibility.
     For known or suspect carcinogens such as benzene, EPA has taken the
position that, because carcinogenicity is considered to be a
nonthreshold toxic effect, water quality criteria for such substances
should be set at zero for the maximum protection of human health  (EPA
1980a, 1980b). However, because zero levels may not be attainable, EPA
has followed a procedure of estimating the concentrations of a substance
in ambient waters which would correspond to incremental increases  in
lifetime cancer risks of 10-5, 10'6, and 10'7. For benzene the
corresponding levels were estimated to be 6.6, 0.66, and 0.066 Mg/L,
respectively (EPA 1980b).
     Because of benzene's known carcinogenicity,  EPA's Office  of
Drinking Water has set the maximum contaminant level goal (MCLG)  for
benzene in drinking water at zero  (EPA 1984c; EPA 1985c). The  MCLG is a.
nonenforceable guideline. The Office of Drinking  Water has also
estimated the benzene concentrations in drinking  water that would
correspond to carcinogenic risks of 10'4, 10'5, and  10'6. These values
are 68, 6.8, and 0.68 MgA. respectively  (EPA 1985a).

-------
                                    Regulatory and Advisory Scacus   L25

     EPA also issues nonenforceable health advisories (HAs) for drinking
water contaminants based on noncarcinogenic health effects. HAs identify
maximum safe drinking water concentrations for specified exposure
periods. The 10-day HA for benzene was reported as 354 jig/L (EPA I985f)
     In 1980, the National Research Council (NRC) derived a suggested
no-adverse-response level (SNARL) for benzene in drinking water. Using
the data of Wolf et al. (1956) for the occurrence of leukopenia and
erythrocytopenia in rats exposed to benzene,  NRC calculated a 7-day
SNARL of 12.6 mg/L (NRC 1980a).
     Food.  No information was found concerning advisory guidelines for
limiting benzene levels in foodstuffs.

9.2.3  Data Analysis

9.2.3.1  Care inogenic potency
     Benzene has been classified by EPA in Group A - Human carcinogen
(EPA 1986, EPA 1988b).  This category is for agents for which there is
sufficient evidence to support a causal association between exposure and
cancer in humans and, in the case of benzene, a sufficient and
supporting level of animal evidence. According to IARC (1982), there is
"sufficient evidence" that benzene is carcinogenic to man and "limited
evidence" that benzene is carcinogenic in experimental animals
(Group A).
     The EPA has developed quantitative unit cancer risk estimates for a
number of known or suspect carcinogens. The methodology for developing
these estimates is given in Anderson (1983).  For benzene, EPA (1986)
derived a cancer risk estimate from data obtained in epidemiologic
studies on workers exposed to benzene vapors (Rinsky et al. 1981, Wong
et al.  1983, Ott et al. 1978). Using an average derived from the
application of several mathematical models, a combined risk estimate of
2.6 x 10*2 was calculated for an exposure to 1 ppm benzene [equivalent
to 0.029 (mg/kg/day)"^ for a lifetime exposure]. For this estimated
potency value, the air concentration associated with an excess risk of
10*5 was derived as 10*5 divided by 2.6 x 10'2 or 3.846 x 10'4 ppm. An
evaluation of Rinsky (1987) data and its impact on cancer potency has
not been developed by EPA as yet.

9.3  STATE
     (Regulations and advisory guidance from the states were still being
compiled at the time of printing.)

9.3.1  Regulations
     California currently lists benzene as a hazardous substance subject
to the minimum standards for management of hazardous and extremely
hazardous wastes (Hazardline 1987).

9.3.1.1  Media-specific

     Air.  No information was found.

-------
126   Section 9
     Water.  In Florida, Che maximum permitted benzene concentration in
drinking water and the most common class of groundwaters is 1
(1 ppb) (FPC 1986).
     Food.  No information was found.
                      •   *
9.3.2  Advisory Guidance
     No information was found.

-------
                                                                     127
                             10. REFERENCES
AAI. 1980. Alliance of American Insurers. Handbook of organic industrial
solvents. 5th ed.

ACGIH (American Conference of Governmental Industrial Hygienists). 1986.
TLVs. Threshold Limit Values for Chemical Substances in the Work
Environment Adopted by ACGIH, with Intended Changes for 1986-1987, and
Biological Exposure Indices for 1986-1987. Cincinnati, OH.

Adams EM, Irish DD, Spencer HC, Rowe VK. 1941. Response of rabbit skin
to compounds reported to have caused acneform dermatitis. Ind Med 2:1-4.
(as reported in Wolf et al. 1956).

Aksoy M. 1978. Benzene and leukaemia. IARC Monograph 29 Lancet i:441.

Aksoy M. 1980. Different types of malignancies due to occupational
exposure to benzene: A review of recent observations in Turkey.  Environ
Res 23:181-190.

Aksoy M. 1981. Problems with benzene in Turkey. Regul Toxicol Pharmacol
1:147-155 (as reported in CA Snyder 1987).

Aksoy M. 1985a. Malignancies due to occupational exposure to benzene.
Am J Ind Med 7:395-402.

Aksoy M. 1985b. Benzene as a leukemogenic and carcinogenic agent. Am J
Ind Med 8:9-20.

Aksoy M, Dincol K. Erdem S, Akgun T, Dincol G. 1972. Details of blood
changes in 32 patients with pancytopenia associated with long-term
exposure to benzene. Br J Ind Med 29:56-64.

Aksoy M, Dincol K, Akgun T, Erdem S, Dincol G. 1971. Haematological
effects of chronic benzene poisoning in 217 workers. Br J Ind Med
28:296-302 (as reported in Goldstein 1977).

Aksoy M, Erdem S. 1978. Followup study on the mortality and the
development of leukemia in 44 pancytopenic patients with chronic benzene
exposure. Blood 52:285-292.
*Key studies.

-------
128   Section 10

* Aksoy M, Erdem S. Erdogan G, Dlncol G. 1976.  Combination of genetic
factors and chronic exposure to benzene in the  aetology of leukaemia
Hum Hered 25:149-153.

Aksoy M, Erdem S, Dincol G. 1974a. Leukemia in  shoe-workers exposed
chronically to benzene. Blood 44:837.

Aksoy M, Erdem S, Dincol K, Hepyuksel T, Dincol G.  1974b.  Chronic
exposure to benzene as a possible contributory  factor in Hodgkin's
disease. Blut 28:293-298.

Aksoy M, Erdem S, Erdogan G, Dincol G. 1976.  Hum Hered 26:149.

Alderson M, Rushton L. 1982. Mortality patterns in eight UK oil
refineries. Ann NY Acad Sci 381:139-145.

Anbar M, Neta P. 1967. A compilation of specific bimolecular rate
constants for the reactions of hydrated electrons,  hydrogen atoms and
hydroxyl radicals with inorganic and organic  compounds in aqueous
solutions. Int J Appl Radiat Isot 18:493-523.

Anderson D, Richardson CR. 1981. Issues relevant to the assessment of
chemically induced chromosome damage in vivo  and their relationship to
chemical mutagenesis. Mutat Res 90:261-272.

Anderson EL. 1983. Quantitative approaches in use to assess cancer risk.
Risk Anal 3:277-295.

Andrews LS, Snyder R. 1986. Toxic effects of  solvents and vapors. In:
Casarett LJ, Doull J, ed. Toxicology, the Basic Science of Poisons, 3rd
ed. Chapter 20. New York: McMillan Publishing Co, p. 641.

Angerer J, Szadkowski D, Manz A, Pett R, Lehnert G. 1973.  Chronic
exposure to organic solvents at the working site. I. Gas-chromatographic
determination of benzene and toluene in the air and in the vapor phase
of blood samples. Int Arch Arbeitsmed 31(1):1-8.

Anonymous. 1986. Sex difference in the micronucleus test.  The
Collaborative Study Group for the Micronucleus  Test. Mutat Res 172:151-
163.

Antoine SR, Delon IR, 0'Dell-Smith RM. 1986.  Environmentally significant
volatile organic pollutants. Bull Environ Contain Toxicol 36(3):364-371.

Appuhn E, Goldeck H. 1957. Fruh-und spatschaden der blutbidung durch
benzol und seine homologen. Arch Gewerbepathol  Gewerbehyg 15:399-428  (as
reported in Goldstein 1977, EPA 1980b).

Arp EU, Wolf PH, Checkoway H. 1983. Lymphocytic leukemia and exposures
to benzene and other solvents in  industry. J Ind Med 25:598-602  (as
reported in Kalf et al. 1987).

-------
                                                        References    129
Ash, UN, Lynch JR. 1971. The evaluation of gas detector cube >y"">-
Benzene. AnTlnd Hyg Assoc J 32:410-411 (as reported in Rinsky.  et al ,

1981).

Auer  (No date)  Auer-prufrorchen fur Auer-Toxiraeter und Auer-Gas
tester  Auergesselschaft, 1000 Berlin 65 (west) (as reported in
Verschueren 1983).

Axelsson G. Luetz C, and Rylander R. 1984. Exposure to "Ivents and
outcome of pregnancy in university  laboratory employees. Br J Ind Med
41:305-312.

Baarson K  Snyder CA,  Albert  RE.  1984. Repeated exposures of C57B1 mice
lolOwl' inSaled benzene markedly  depressed  "^"P"1"" "^I*
formation. Toxicol  Lett 20:337-342  (as reported in  C.A.  Snyder 1987)
 Appl Pharmacol 64:393-404.




 16:275-282.
                             ^
 Mutat Res 144:193-196.
 Barlow SM, Sullivan FM.  1982. Reproductive Hazards of Industrial
 Chemicals. London: Academic, pp.  83-103.




  EPA/600/8/032a-86.





  Elsevier, p. 347-352.
  BM.lt RC. 1974. Letter: Blood benzene stability in plastic containers.
  Clin-Chem 20(11) : 1477-1478.

  Battennann G.  1986.  Decontamination  * P^j^'™ Van den Brink,
  biodegradation.  In:  Contam Soil  Int  TOO  Conf.  AssinK JW
  WJ  eds   Dordrecht,  Netherlands:  Nijhoff,  pp.  711  ILL  ^as  «P
  Chem Abst 104(24): 212909U) .

-------
130   Section 10

Baxter HG, Blakemore R, Moore JP, Coker DT.  1980.  The measurement of
airborne benzene vapour. Ann Occup Hyg 23:117-132  (as reported in IARC
1982).

Bergman K. 1979. Whole-body autoradiography and allied tracer techniques
in distribution and elimination studies of some organic solvents. Scand
J Work Environ Health 5:29-92 (as reported in Dempster et al. 1984).

Berkley R, Scott OR, Hedgecoke R. 1984. Performance optimization of
TALMS. American Chemical Society Meeting,  Miami,  Florida. April 1984 (as
reported in Hadeishi et al. 1985).

Bernard J, Bassett A. 1946. Results of an inquiry  on benzolism in the
aeronautic industry. Sang 17:120 (as reported in Goldstein 1977).

Blackwood TR, Ochsner JC, Zaegler LD. 1979.  Status Assessment of Toxic
Chemicals: Benzene (PB80-146335). Prepared for the U.S. Environmental
Protection Agency by Monsanto Research Corp.  Springfield, VA: National
Technical Information Center.

Blanchard RD, Hardy JK. 1986. Continuous monitoring device for the
collection of 23 volatile organic priority pollutants. Anal Chem
58(7):1529-1532.

Blaney L. 1950. Early detection of benzene toxicity. Ind Med 19:227-228
(as reported in Goldstein 1977)

Blank IH, McAuliffe DJ. 1985. Penetration of benzene through human skin.
J Invest Dermatol 85:522-526.

Blattner WA, Strober W, Muchmore AV, Blaese RM, Broder S, Fraumen JF.
1976. Familial chronic lymphocytic leukemia.  Ann Int Med 84:554-557.

Bock FG, King DW. 1959. A study of the sensitivity of the mouse
forestomach toward certain polycyclic hydrocarbons. J Natl Cancer Inst
23:833-839.

Bolcsak LE, Nerland DE. 1983. Inhibition of erythropoiesis by benzene
and benzene metabolites. Toxicol Appl Pharmacol 69:363-368.
                                                           V.
Bond GG, McLaren EA, Baldwin CL, Cook RR. 1986. An update of mortality
among chemical workers exposed to benzene. Br J Ind Med 43:685-691.

Bousser JR, Neude R, Fabre A. 1947. Un cas ALL hemopathic benzolique
tres relardee un type de lymphosarcome. Bull Mem Soc Med Hop Paris
63:1100  (as reported in Aksoy 1985a).

Bozzelli JW, Kebbekus BB. 1982. A study of some aromatic and halocarbon
vapors in the ambient atmosphere of New Jersey. J Environ  Sci Health
A17(5):6 93-711.

-------
                                                        References   131

Bradley EB, Frenzel CA. 1970. On the exploitation of Laser raman
spectroscopy for the detection and identification of molecular water
pollutants. Water Res 4:124-128.

Bradley MO  1985  Measurement of DNA single-strand breaks by alkaline
elution in'rat hepatocytes. In: Progress in Mutation Research 5  Ashby
j  de Serres FJ  Draper M, Ishidate M Jr, Margolin BH, Matter BE, Shelby
MD  eds  Evaluation of Short-Term Tests for Carcinogens: Report of the
International Program on Chemical Safety Collaborative Study on in vitro
Assays. Amsterdam: Elsevier, pp. 353-357.

Braier L,  Francone M.  1950. Title not given. Arch Mai Prof 11:367 (as
reported in Sandmeyer  1981).

Braier L,  Levy A, Dror K,  Pardo A.  1981. Benzene in blood and Phenol in
urine  in monitoring benzene  exposure  in  industry. Am J  Ind Med 2:119-
123.

Branson DR. 1978. Predicting the  fate of chemicals  in the aquatic
environment from laboratory data. ASTM STP 657, pp.  55-70.

Brass  HJ,  Feige  MA,  Halloran T,  Mello JW,  Munch D,  Thomas RF.  1977.  The
national monitoring survey:  samplings and  analyses  for  purgeable organic
compounds. Drinking Water Quality Enhancement Source Protection,  pp.
393-416.

Brief RS,  Lynch J,  Bernath T, Scala RA.  1980. Benzene in the workplace.
Am Ind Hyg Assoc J  41:616-623.

 Browning E.  1965. Toxicity and metabolism of industrial solvents. New
 York:  Elsevier Scientific Publishing Co.,  pp 3-65 (as reported in
 Infante and White 1983) .

 Bryce-Smith D, Gilbert A. 1976. The organic photochemistry of benzene.
 I. Tetrahedron 32:1309-1326  (as reported  in EPA 1983).

 Brzecki A, Misztel S, Kostolowski R. 1973. Title not given. Ann Acad Med
 Lodz 14(1):55 (as reported  in Sandmeyer 1981).

 Budnick LD, Sokal DC, Falk  H,  Logue  JN, Fox  JM. 1984.  Cancer  and birth
 defects near  the Drake superfund site,  Pennsylvania. Arch Environ  Health
 39:409-413.

 Bull  RJ,  Robinson  M,  Laurie RD.  1986. Association  of carcinoma  yield
 with  early papilloma  development in SENCAR mice. Environ Health Perspect
 68:11-17.

 Burdette  WJ.  Strong LC.  1941. Comparison of methylsalicylate and benzene
 as  solvents  for methyocholanthrene. Cancer Res 1:939-941.

 Burmaster DE. 1982.  The new pollution-groundwater contamination. Environ
 24:6-13,  33-36 (as reported in CHEMFATE 1987).

-------
 132    Section 10

 Byron  PA,  Goer P,  Girard R,  Gentihomme 0, Revol L. 1969. Acute
 erythromyelosis with benzene etiology. J Med Lyon 50:757-759  (as
 reported in IARC  1982).

 Caprotti M,  et al.  1962.  Ulcers  in  the mucous membrane of the colon in
 benzene  poisoning:  Clinical  and  radiological study. Lavoro Umano 14:445
 (as  reported in EPA 1980b).

 Carpenter C.  Shaffer C,  Weil C,  Smyth H. 1944. Studies on the inhalation
 of 1:3 butatiene;  with comparison of its narcotic effect with benzol,
 toluol,  and styurene and a note  on  the elimination of styrene by the
 human. J Ind Hyg Toxicol 26:69-78.

 C&EN.  1987a.  Chemical and Engineering News.  Benzene.  February 9  1987
 p. 11.

 C&EN.  1987b.  Chemical and Engineering News.  Top 50 chemicals  April 13
 1987, p.  21.

 CEH. 1986.  Chemical  Economic Handbook. Section 300.7000A- 300.7003S
 Menlo Park,  CA: SRI  International.

 CEH. 1987.  Chemical  Economic Handbook. Section 300.70021- 300.7003S
 Menlo Park,  CA: SRI  International.

 Chang IW.  1972. Study on the threshold limit value of benzene and early
 diagnosis  of benzene  poisoning. J Cath Med Coll 23:429-434.

 CHEMFATE.  1987. Benzene.  Syracuse Research Corporation.

 CHEM. ABSTR.  1977-1981.  Chemical Abstracts.  10th Collective Index.
 1977-1987. Chemical Abstracts Service, American Chemical Society.

 CHEMLINE.  1987. Chemical  Abstracts. Online file (also reported in HSDB
 1987).

 Chiou CT,  Freed VH,  Schmedding DW, Kohnert RL. 1977.  Partition
 coefficient  and bioaccumula.tion of selected organic chemicals. Environ
 Sci Technol  11:475-478.
                                                          t
 Chiou CT,  Schmedding  DW,  Manes M. 1982. Partitioning of organic
 compounds  in  octanol-vater systems. Environ Sci Technol 16:4-9.

 Choy WN, MacGregor JT, Shelby MD, Maronpot RR. 1985.  Induction of
micronuclei by benzene in B6C3F]. mice: Retrospective analysis of
 peripheral blood smears  from the NTP carcinogenesis bioassay. Mutat Res
 143:55-59.

Clark AI, Mclntyre AE, Lester JN, Perry R. 1984.  A comparison of photo-
 ionization detection  gas  chromatography with a Tenax GC sampling tube
procedure  for the measurement of aromatic hydrocarbons in ambient air.
 Int J Environ Anal Chem  17(3-4):315-326.

-------
                                                        References   133

Coate WB, Hoberman AM, Durloo RS. 1984. Inhalation teratology study of
benzene in racs. Adv Mod Environ Toxicol 6:187-198.

Cole P, Merletti F. 1980. Chemical agents and occupational cancer. J
Environ Pathol Toxicol 3:399-417.

Colenutt BA, Davies DN. 1980. The sampling and gas chromatographic
analysis of organic vapors in landfill sites. Int J Environ Stud 15:25-
32 (as reported in IARC 1982).

Colenutt BA, Thorburn S. 1980. Gas chromatographic analysis of trace
hydrocarbon pollutants in water samples. Int J Environ Stud 15:25-32.

Conkle JP, Camp BJ, Welch BE. 1975.  Trace composition of human
respiratory gas. Arch Environ Health 30:290-295.

Cornish HH, Ryan RC. 1965. Metabolism of benzene in nonfasted, fasted,
and aryl-hydroxylase inhibited rats. Toxicol Appl Pharmacol 7:767-771
(as reported in IARC 1982).

Cox RA, Derwent RG, Williams MR. 1980. Atmospheric photoxidation
reactions. Rates, reactivity, and mechanism for reaction of organic
compounds with hydroxyl radicals. Environ Sci Technol 14:5761.

CPSC. 1981. U.S. Consumer Product Safety Commission. Benzene-containing
consumer products: withdrawal of proposed rule. Fed Regist 46:27910-
27911.

Crebelli R, Bellincampi D, Conti G,  Conti L, Morpurgo G, Carere A. 1986
A comparative study on selected chemical carcinogens for chromosome
malsegregation, mitotic crossing-over and forward mutation induction in
Aspergillus nidulans. Mutat Res 172:139-149.

* Cronkite EP. 1986. Benzene hematotoxicity and leukemogenes is.  Blood
Cells 12:129-137.

Cronkite EP, Bullis J, Inoue T, Drew RT. 1984. Benzene inhalation
produces leukemia in mice. Toxicol Appl Pharmacol 75:358-361.

Cronkite EP, Drew RT, Inoue T, Bullis JE. 1985. Benzene'hematotoxicity
and leukemogenesis. Am J Ind Med 7:447-456.

Cronkite EP. Inoue T. Carsten AL, Miller ME, Bullis JE, Drew RT.  1982.
Effects of benzene inhalation on marine pluripotent stem cells. J
Toxicol Environ Health 9:411-421.

Crump KS, Allen BC. 1984. Quantitative Estimates of Risk of Leukemia
from Occupational Exposure to Benzene. Washington, DC: U.S. Occupational
Safety and Health Administration.

-------
134   Section 10

CSSTT Study. 1985. Comparative studies on short-term tests. In: Progress
in Mutation Research 5. Ashby J, de Serres FJ,  Draper M, Ishidate M Jr,
Margolin BH, Matter BE, Shelby MD, eds. Evaluation of Short-Term Tests
for Carcinogens: Report of the International Program on Chemical Safety
Collaborative Study on in vitro Assays. Amsterdam:  Elsevier.

Curletto R, CLconali M. 1962. Hematological disorders in benzene
poisoning. Med Lavoro 53:505-546 (as reported in Goldstein 1977).

Dean BJ.  1978. Genetic toxicology of benzene, toluene, xylenes and
phenols.  Mutat Res 47:75-97.

Dean BJ.  1985. Recent findings on the genetic toxicology of benzene,
toluene,  xylenes and phenols. Mutat Res 154:153-181.

Decoufle P, Blattner WA, Blair A. 1983. Mortality among chemical workers
exposed to benzene and other agents. Environ Res 30:16-25.

De Flora S, Zanacchi P, Camoirano A, Bennicelli C,  Badolati GS. 1984.
Genotoxic activity and potency of 135 compounds in the Ames reversion
test and in a bacterial DNA-repair test. Mutat Res 133:161-198.

DeGowin RL. 1963. Benzene exposure and aplastic anemia followed by
leukemia 15 years later. J Am Med Assoc 185:748-751 (also reported in
IARC 1982).

Delore P, Borgamano J. 1928. Leucemie aique en cours d'intoxication
benzenique. J Med Lyon 9:227-233 (as reported in IARC 1982).

Dempster AM, Evans HL, Snyder CA. 1984. The temporal relationship
between behavioral and hematological effects of inhaled benzene. Toxicol
Appl Pharmacol 76:195-203.

Desoille H, Albahary C, Philbert M. 1965. Incidences hormonales sur  le
benzenism chronique de la rate. Influence de la gestation sur  le taux
des globules blancs et des  globules rouges de la rate soumise  ou non a
une intoxication benzenique moderee pendant toute la duree de  la
gravidite. Arch Mai Prof Med Trav Secur Soc 26:205-220  (as reported  in
Barlow and Sullivan 1982).

Desoille H, Philbert M. Albahary C. 1967. Hormonal  incidences  of chronic
benzene poisoning in guinea pigs. Influence of gestation  on white and
red corpuscle levels in guinea  pigs submitted or not  to moderate benzene
poisoning during  the entire gestation  period. Arch  Mai  Prof Med Trav
Secur Soc 28:329-339 (as  reported in Barlow and Sullivan  1982).

DIALOG (SSIE Current Research or  Federal Research in  Progress).  1987.
Lockheed database system. Computer  printout  retrieved 7/87.

Diaz M, Reiser A, Braier  L,  Diez J. 1980.  Studies on  benzene
mutagenesis.  I. The micronucleus  test.  Experientia  36:297-299.

-------
                                                        References   135

DiGulielfflo G, lannaccone A. 1958. Inhibition of mitosis and regressive
changes of erytroblasts in acute erythropathy caused by occupational
benzene poisoning. Arch Haematol 19:144-147 (as reported in IARC 1982).

Ding X-J, Li Y, Ding Y, Yang H-Z. 1983. Chromosome changes in patients
with chronic benzene poisoning. Chinese Med J (Peking Engl Ed) 96:681-
685.

Dobashi Y. 1974. Influence of benzene and its metabolites on mitosis of
cultured human cells. Japan J Ind Health 16:453-461.

Dorfman LM, Adams GE. 1973. Reactivity of the hydroxyl radical in
aqueous solutions. NSRD-NBS-46. Washington DC: National Bureau of
Standards. 51 pp (as reported in CHEMFATE 1987).

Douglas GR, Blakey DH, Liu-Lee VU, Bell RDL, Bayley JM. 1985. Alkaline
sucrose sedimentation, sister-chromatid exchange and micronucleus assays
in CHO cells. In: Progress in Mutation Research 5. Ashby J, de Serres
FJ, Draper H, Ishidate H Jr, Margolin BH, Matter BE, Shelby MD, eds.
Evaluation of Short-Term Tests for Carcinogens: Report of the
International Program on Chemical Safety Collaborative Study on in vitro
Assays. Amsterdam: Elsevier, pp. 359-366.

Dowty BJ, Laseter JL, Storer J. 1976. The transplacental migration and
accumulation in blood of volatile organic constituents. Pediatr Res
10:696-701.

Drager. (No date). Gasspurgerat. 3. Dragerwerk AG, 24 Lubeck 1 Postfach
1339 (as reported in Verschueren 1983).

* Drew RT, Fouts JR. 1974. The lack of effects of pretreatment with
phenobarbital and chlorpromazine on the acute toxicity of benzene in
rats. Toxicol Appl Pharmacol 27:183-193.

Drogichina EA, Zorina LA, Gribova IA. 1971. Title not given. Gig Tr Prof
Zabol 15(5):18 (as reported in Sandmeyer 1981).

Du Pont de Nemours & Co. (No date). Concentrations of liquids, gases and
vapors measured with the Du Pont 400 photometric analyzer. Bulletin 5A.
DuPont de Nemours & Co. (as reported in Verschueren 1983).

Dutkiewicz T, Tyras H. 1968. The quantitative estimation of  toluene skin
absorption in man. Int Arch Gewerbepath Gewerbehyg 24:253-257.

Dutkiewicz T, Tyras H. 1967. A study of skin absorption of ethylbenzene
in man. Br J Ind Med 24:330-332.

Duvoir MM, Derobert L. 1942. Les hemopathies benzoliques retardees. Sang
15:267-272 (as reported in Goldstein 1977).

Duvoir MR, Fabre A, Derobert L. 1946.  [The  significance of benzene  in
the bone marrow in the course of benzene blood  diseases. ] Arch Mai  Prof
7:77.

-------
136   Section 10

Dyksen JE, Hess AF, III. 1982. Alternatives for controlling organics in
ground water supplies. J Am Water Works Assoc 74:394-403.

Emile-Weil MP. 1932. La leucimie post-benzolique. Bull Mem Soc Med Hop
Paris 47:193-198 (as re.por.ted in Goldstein 1977).

EPA (Environmental Protection Agency). 1975.  Supplement to Development
Document: Hazardous Substances Regulations, Sect. 311, FWPCA.
Washington, DC: Environmental Protection Agency.  EPA
440/9-75-009 (as reported in HSDB 1987)

EPA (Environmental Protection Agency). 1980a.  Water quality criteria
documents; availability. Fed Regist 45:79318-79324.

* EPA (Environmental Protection Agency). 1980b.  Ambient Water Quality
Criteria for Benzene. Washington, DC: Environmental Protection Agency,
pp. C-16-C-36.

EPA (Environmental Protection Agency). 1981.  National emission standard
for hazardous air pollutants; benzene fugitive emissions. US Code Fed.
Regul.,  Title 40, Part 61; Fed Regist. 46(2),  pp. 1165-1193 (as reported
in IARC 1982).

EPA (Environmental Protection Agency). 1982.  Methods for the Organic
Chemical Analysis of Municipal and Industrial Wastewater--Method 602.
Cincinnati, OH: Environmental Protection Agency,  Environmental
Monitoring and Support Laboratory. EPA-600/4-82-057. pp. 601-1-10.

EPA (Environmental Protection Agency). 1983.  Hazard Profile for Benzene.
Environmental Criteria and Assessment Office,  Cincinnati, OH.  Draft
report.

EPA (Environmental Protection Agency). 1984a.  Health Effects Assessment
for Benzene. Office of Health and Environmental Assessment. EPA/540/1-
86-037.

EPA (Environmental Protection Agency). 1984b.  Guidelines for
establishing test procedures for the analysis of pollutants under the
Clean Water Act. Fed Regist 49:43234, 43255-56,  43272-43280, 43373-
43384, 43407-43415.

EPA (Environmental Protection Agency). 1984c.  National primary drinking
water regulations, volatile synthetic organic chemicals; proposed rule
making.  Fed Regist 49:24330-24355.

EPA (Environmental Protection Agency). 1985a.  Drinking Water Criteria
Document on Benzene. Final draft. Washington,  DC: Office of Drinking
Water, Environmental Protection Agency. PB86-118122.

EPA (Environmental Protection Agency).  1985b. Pesticide chemicals
category effluent limitations guidelines, pretreatment standards, and
new source performance standards. Fed Regist 50(193):40707.

-------
                                                         References   137

 EPA (Environmental Protection Agency). 1985c. National primary drinking
 water regulations: volatile synthetic organic chemicals  final rule   Fed
 Regist 50:46880-46901.

 EPA (Environmental Protection Agency). 1985d. National primary drinking
 water regulations: volatile synthetic organic chemicals.  Fed Resist
 50:46901-46933.

 EPA (Environmental Protection Agency). 1985e. Exemptions  from
 tolerances.  Code of Federal Regulations,  Title 40.  Part 180.  Section
 180.1001,  Subpart D,  p.  355,  July 1,  1985.

 EPA (Environmental Protection Agency). 1985f. Health Advisory for
 Benzene. Washington,  DC:  Office of Drinking Water,  September  30, 1985.

 * EPA (Environmental  Protection Agency).  1986.  Evaluation of  the
 Potential  Carcinogenicity of  Benzene.  Review Draft. Carcinogen
 Assessment Group,  Office  of Health and Environmental Assessment
 OHEA-C-073-29.

 EPA (Environmental Protection Agency).  1987a.  National  primary drinking
 water regulations:  Synthetic  organic  chemicals; monitoring for
 unregulated  contaminants.  Final rule.  Fed Regist  52:25690-25717.

 EPA (Environmental Protection Agency).  1987b.  Volatile  Organic Chemicals
 Methods and  Monitoring Document.  Washington,  DC:  Office of Water,
 Environmental Protection  Agency.

 EPA (Environmental Protection Agency).  1988a.  Analysis  of Clean Water
 Act Effluent Guidelines pollutants. Summary of the  Chemicals  Regulated
 by  Industrial Point Source  Category,  40 CFR Parts 400-475. Washington,
 DC:  Office of Water,  Environmental  Protection Agency. Draft.

 EPA (Environmental  Protection Agency).  1988b.  Integrated  Risk
 Information System, Benzene Part  II, March  1,  1988.

 Erdogan G,  Aksoy M. 1973. Cytogenetic  studies  in  thirteen patients with
 pancytopenia and leukemia associated with long-term exposure  to benzene.
 New Istanbul Contrib  Clin Sci  10(4):230-247.
                                                        V.
 Erexson GL, Wilmer  JL, Kligerman AD.  1985.  Sister chromatid exchange in
 human lymphocytes  exposed to benzene and  its  metabolites  in vitro.
 Cancer Res 45:2471-2477.

 Erexson GL, Wilmer  JL, Steinhagen WH,  Kligerman AD. 1986. Induction of
 cytogenetic damage  in rodents  after short-term  inhalation of  benzene.
 Environ Mutagen 8:29-40.

 Erf LA, Rhoads CP.  1939. The hematological  effects of benzene  (benzol)
poisoning.  J Ind Hyg  Toxicol  20(8):421-435  (as reported in Goldstein
 1977).

-------
138   Seccion 10

Ewlng BB, Chain ESK. Cook JC, Evans CA, Hopke PK,  Perkins, EG. 1977.
Monitoring Co Detect Previously Unrecognized Pollutants in Surface
Waters. Appendix: organic analysis data. Washington,  DC:  Environmental
Prelection Agency. EPA-560/6-77-015A (as reported in CHEMFATE 1987).

Favre-Gilley M, Bruel Mile. 1948. Syndrome hemorragique benzolique avec
simple alteration morphologiques des plaquettes.  De putilite d'examen
systematique des plaquettes sur lame chez les ouvriers exposes au
benzol. Arch Mai Prof 91:274-277 (as reported in Goldstein 1977).

Fel'dt EG. 1985. Evaluation of the mutagenic hazards of benzene and some
of its derivatives. Gig Sanit 7:21-23.

Fentiman AF, Neher MB, Kinzer GW, Sticksel PR,  Coutant RW. 1979.
Battelle Columbus Environmental Monitoring Benzene. EPA 560/679/006.
Washington, DC: Office of Toxic Substances, Environmental Protection
Agency.

Ferrario JB, Lawler GC, Delon IR, Laseter JL. 1985. Volatile organic
pollutants in biota and sediments of Lake Pontchartrain.  Bull Environ
Contam Toxicol 34(2): 246-255.

Fishbeck WA, Langner RR, Kociba RJ.  1975. Elevated urinary phenol levels
not related to benzene exposure. Am Ind Hyg Assoc J 36(11):820-824.

Folin 0, Denis W. 1915. The excretion of free and conjugated phenols and
phenol derivatives. J Biol Chem 22:309-320.

Forni A. 1979. Chromosome changes and benzene exposure. A review. Rev
Environ Health 3:5-17.

Forni A, Moreo L. 1967. Cytogenetic studies in a case of benzene
leukemia. Eur J Cancer 3:251-255.

Forni A, Moreo L. 1969. Chromosome studies in a case of benzene-induced
erythroleukemia. Eur J Cancer 5:459-463 (as reported in IARC 1982).

Forni A, Pacifico E, Limonta A. 1971a.  Chromosome studies in workers
exposed to benzene or toluene or both.  Arch Environ Health 22:373-378.

Forni AM, Cappellini A, Pacifico E,  Vigliani EC.  1971b. Chromosome
changes and their evolution in subjects with past exposure to benzene.
Arch Environ Health 23:385-391.

Franz TJ. 1984. Percutaneous absorption of benzene. In: Advances in
Modern Environmental Toxicology, Vol. VI. Applied Toxicology of
Petroleum Hydrocarbons. MacFarland HN et al., eds.  Princeton, NJ:
Princeton Scientific Publishers, Inc.

FPC/API. 1986. Florida Petroleum Council. Benzene  in Florida
Groundwater. An Assessment of the Significance to Human Health.  Florida
Petroleum Council, Division of the American Petroleum Institute. 161 pp.

-------
                                                        References   139

Frash VN,  Yushkov BG, Karaulov AV, Skuratov VL.  1976.  Mechanism of
action of benzene on hematopoiesis (investigation of hematopoietic stem
cells). Bull Exptl Biol Med 82(7) :985-987.

Funes-Cravioto F, Zapata-Gayon C. Kolmodin-Hedman B, 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: 322-32S.

Gad-el-Karim MM, Ramanujam VMS, Legator MS. 1985. Trans,trans-muconic
acid, an open chain urinary metabolite of benzene in mice.
Quantification by high-pressure liquid chromatography. Xenobiotica
15:211-220.

Gad-el Karim MM, Sadagopa Ramanujam VM, Legator MS. 1986. Correlation
between the induction of micronuclei in bone marrow by benzene exposure
and the excretion of metabolites  in urine of CD-I mice. Toxicol Appl
Pharmacol 85:464-477.

Gaffney JS, Levin SZ. 1979. Predicting gas phase organic molecular
reaction rates using linear free-energy correlations. I. 0(3P) and OH
addition and abstraction reactions. Int J Chem Kinet XI:1197-1209.

Gaido K, Wierda D. 1984. In vitro  effects of benzene metabolites on
mouse bone marrow stromal cells.  Toxicol Appl Pharmacol 76:45-55.

Galavotti B, Troisi  FM.  1950.  Erythroleukemia myelosis  in benzene
poisoning. Br J Ind  Med  7:79-81  (as reported in  IARC  1982).

Gallinelli R. 1966.  Acute paramyeloblastic  leukosis due  to chronic
benzene poisoning in a student mosaicist. Med Lav  57:257-261  (as
reported in Goldstein 1977).

Geist  CR, Kelly LD,  Schoenheit CM,  Praed JE. 1983.  Learning  impairments
following postnatal  exposures  to benzene.  Percept  Motor Skills  57:1083-
1086.

* Gerarde HW. 1959.  lexicological studies  on hydrocarbons. Am Med Assoc
Arch  Ind Health  19:403-418.

* Gerarde HW. 1960.  Toxicology and biochemistry of aromatic
hydrocarbons. Elsevier  Publishing Co.:  London,  pp. 52-53.

Gerarde HW.  1963. The  aromatic hydrocarbons.  In: Patty's Industrial
Hygiene and Toxicology,  Vol.  2,  2nd rev.  ed.,  Fassett DW and Irish DD,
eds.  New York:  Interscience Publishers,  pp. 1219-1225.

Gerner-Smidt P,  Friedrich U.  1978. The mutagenic effect of benzene,
toluene and xylene  studied by the SCE technique. Mutat Res 58:313-316.

Ghantous  H,  Danielsson BRG. 1986. Placental transfer and distribution of
 toluene,  xylene and benzene,  and their metabolites during gestation in
mice.  Biol Res  Pregnancy Perinatol. 7:98-105.

-------
140   Section 10

Ghimentl G, Gal11 M, Galli E. 1978. Extraction and determination of
solvents In biological samples. Part 1.  o Ann 1st Super Sanit 14:5 (as
reported in IARC 1982).

Gibson DT. 1977. Biodegradaion of aromatic petroleum hydrocarbons. In:
Fate and Effects of Petroleum Hydrocarbons in Marine Ecosystems and
Organisms. Wolfe DA, ed. New York: Pergamon,  pp.  36-46.

Gibson DT. 1980. Microbial metabolism.  In: The Handbook of Environmental
Chemistry. Hutzinger 0, ed. New York:  Springer-Verlag,  pp. 161-192.

Gilbert D, Byrne M, Harris J, Steben W,  Woodruff  C.  1982.  An Exposure
and Risk Assessment for Benzene. Environmental Protection Agency.  EPA-
440/4-85-006. PB 85-212017, pp. 3-1 -  3-32.

Gill DP, Ahmed AE. 1981. Covalent binding of  [14C]benzene to cellular
organelles and bone marrow nucleic acids. Biochem Pharmacol 30:1127-
1131.

Gill DP, Jenkins VK, Kempen RR, Ellis S. 1980. The importance of
pluripotential stem cells in benzene toxicity. Toxicology 16:163-171.

Girard R, Revol L. 1970. The incidence of exposure to benzene in severe
hemopathies. Nouv Rev fr Hematol 10:477-484 (as reported in IARC 1982,
Goldstein 1977).

Glass LR, Connor TH, Thiess JC, Dallas CE, Hatney TS. 1986. Genotoxic
evaluation of the offgassing products of particle boards. Toxicol Lett
31:75-83.

Glauert HP, Kennan WS, Sattler GL, Pitot HC.  1985. Assays to measure the
induction of unscheduled DNA synthesis in cultured hepatocytes. In:
Progress in Mutation Research 5. Ashby J, de  Serres FJ, Draper M,
Ishidate M Jr, Margolin BH, Matter BE,  Shelby MD, eds.  Evaluation of
Short-Term Tests for Carcinogens: Report of the International Program on
Chemical Safety Collaborative Study on in vitro Assays. Amsterdam:
Elsevier, pp. 371-373.

Gofmekler VA. 1968. Embryotropic action of benzene and formaldehyde
inhalation. Hyg SaniC (USSR) 33(1-3):327-332. (Translated from Gig Sanit
33(3):12-16)

Goguel A, Cavigneaux A, Bernard J. 1967. Benzene leukemias. Bull  Inst
Natl Sante Rech Med 22:421-441  (as reported in IARC 1982).

Goldstein BD. 1986. Biological and ambient monitoring of benzene  in  the
workplace. J Occup Med 28(10):1051-1054.

* Goldstein BD. 1977. Hematotoxicity in humans. J Toxicol Environ Health
Suppl 2:69-105.

-------
                                                        References   141

Goldstein BD. 1985. Risk assessment and risk management of benzene by
the Environmental Protection Agency. In: Risk Quantitation and
Regulatory Policy, Banbury Report 19:293-304.

Goldstein BD. 1986. Critical review of toxic air pollutants--revisited
J Air Pollution Cont Assn 36:367-370.

Goldstein BD, Snyder CA, Laskin S, Bromberg I, Albert RE,  Nelson N.
1982a. Myelogenous leukemia in rodents inhaling benzene. Toxicol Lett
13:169-173.

Goldwater LJ. 1941. Disturbances in the blood following exposure to
benzol. J Lab Clin Med 26:957-973.

Gonasun L, Witmer C, Kocsis J, Snyder R. 1973. Benzene metabolism in
mouse liver microsomes. Toxicol Appl Pharmacol 26:398-406.

Graedel TE. 1978. Hydrocarbons. In: Chemical Compounds in the
Atmosphere. New York: Academic, pp. 50-157.

Green JD, Leong BKJ, Laskin S. 1978. Inhaled benzene fetotoxicity in
rats. Toxicol Appl Pharmacol 46:9-18.

Green JD, Snyder CA, LoBue J, Goldstein BD, Albert RE. 1981. Acute and
chronic dose/response effects of inhaled benzene on multipotential
hematopoietic stem  (CFU-S) and granulocyte/macrophage progenitor
(GM-CFU-C) cells in CD-I mice. Toxicol Appl Pharmacol 58:492-503.

Greenlee WF, Gross EA, Irons RD. 1981. Relationship between benzene
toxicity and the disposition of ^C-labelled benzene metabolites  in the
rat. Chem-Biol Interact 33:285-299.

Greenlee WF, Irons RD. 1981. Modulation of benzene-induced
lymphocytopenia in  the rat by 2,4,5,2',4',5'-hexachlorobiphenyl and
3,4,3',4'-tetrachlorobiphenyl. Chem-Biol Interact 33:345-360.

Grob K, Grob G. 1974. Organic substances in potable water and its
precursor. Part II. Applications  in  the area  of Zurich. J Chromatog
90:303-313.
                                                        '.
Gruenke LD, Craig JC, Wester RC, Maibach HI.  1986. Quantitative analysis
of benzene by selected  ion monitoring/gas  chromatography/mass
spectrometry. J Anal Toxicol 10:225-232.

Guberan E, Kocher P. 1971. Pronostic  lointain de 1'intoxication
benzolique chronique: controle  d'une  population 10 ans  apres
1'exposition. Schweiz Med Worchenscher 101:1789-1790  (as  reported in
Goldstein  1977).

Hadeishi T,  Pollard M,  McLaughlin R,  Koga  M.  1985. Development  of an
optical monitor  for toxic organic  compounds  in air.  EPA/600/4-85-043.
Research Triangle Park, NC:  Environmental  Protection Agency.

-------
142   Section 10

Hager N. 1973. Derivative spectroscopy with emphasis on trace gas
analysis - LSI/Soectrometrlcs, Inc. Anal Chem 45(13) (as reported In
Verschueren 1983).

Haider K, Jagnov G, Kohnen R, Lim SU. 1981. Degradation of chlorinated
benzenes, phenols and cyclohexane derivatives by benzene- and phenol-
utilitizing soil bacteria under aerobic conditions.  In: Decomposition of
Toxic and Nontoxic Organic Compounds in Soils. Overcash MR,  ed.  Ann
Arbor: Ann Arbor Science, pp. 207-223.

Hamilton A. 1922. The growing menace of benzene (benzol) poisoning in
American industry. J Am Med Assoc 78(9):627-630.

Hammers WE, Bosnian HFPM. 1986. Quantitative evaluation of a simple
dynamic headspace analysis technique for non-polar pollutants in aqueous
samples at the ng/kg level. J Chromatogr 360:425-432.

* Hammond JV, Herman ER. 1960. Industrial hygiene features of a
petrochemical benzene plant design and operation. Am Ind Hyg Assoc J
21:173-177.

Hampson RF. 1980. Chemical kinetic and photochemical data sheets for
atmospheric reactions. FAA-EE-80-17.  Washington, DC: U.S. Department of
Transportation (as reported in CHEMFATE 1987).

Hanke J, Dutkiewicz T, Piotrowski J.  1961. The absorption of benzene
through the skin in men. Med Pracy 12:413-426 (as reported in Loden
1986).

Harigaya K, Miller ME, Cronkite EP, Drew RT.  1981. The detection of in
vivo hematotoxicity of benzene by in vitro liquid bone marrow cultures.
Toxicol Appl Pharmacol 60:346-353.

Harkov R, Gianti SJ,  Bozzelli JU, LaRegina JE. 1985. Monitoring volatile
organic compounds at hazardous and sanitary landfills in New Jersey. J
Environ Sci Health A20(5):491-501.

Harland BJ, Whitby FJ, Comber MHI. 1985.  Measurement of volatile
aromatic hydrocarbons in an industrial estuary. Int J Environ Anal Chem
20:295-311.

Harper BL, Sadagopa RamanuJam VM, Gad-el-Karim MM, Legator MS. 1984. The
influence of simple aromatics on benzene clastogenicity. Mutat Res
128:105-114.

Hartwich G, Schwanitz G, Becker J. 1969.  Chromosome anomalies in a case
of benzene leukemia.  Ger Med Mon 14:449-450.

Hay EB. 1964. Exposure to aromatic hydrocarbons in a coke oven by-
product plant. Am Ind Hyg Assoc J 25:3868-391 (as reported in Rinsky et
al. 1981).

-------
                                                        References   143

Hazardline. 1987. Minimum standards for Che management of hazardous and
extremely hazardous wastes. State of California's Administrative Code.
Title 22, Division 4, Chapter 30.

Heath CU. 1983. Field epidemiology studies of populations exposed to
waste dumps. Environ He-alth Perspect 48:3-7.

Herian E. 1973. Umweltforschung uber nichthalogenierte
Kohlenwasswestoffe. Chemische Rundschau 24:10 (as reported in
Verschueren 1983).

Hermann M. 1981. Synergistic effects of individual polycyclic aromatic
hydrocarbons on the mutagenicity of their mixtures. Mutat Res 90:399-
409.

Hernberg S, Savilahti M, Ahlman K, Asp S. 1966.  Prognostic aspects of
benzene poisoning. Br J Ind Med 23:204-209  (as reported in Goldstein
1977, C. Snyder 1987).

Higgins IJ, Best DJ, Hammond RC. 1980. New  findings in methane -
utilizing bacteria highlight their importance in the biosphere and their
commercial potential. Nature 286:561-564.

Hinson JA, Freeman JP, Potter DW, et al. 1985. Mechanism of microsomal
metabolism of benzene to phenol. Mol Pharmacol 27:574-577.

Kite M, Pecharo M, Smith I, Thornton S. 1980. The effect of benzene in
the micronucleus test. Mutat Res 77:149-155.

Holden MW, Smith DL, Smith RN. 1985. Comparison of ambient air sampling
techniques for volatile organic compounds.  Environmental Protection
Agency, EPA 600/4-85-067. PB86-120953/GAR.

Holmberg PC. 1979. Central-nervous-system defects  in children born to
mothers exposed  to organic solvents during  pregnancy. Lancet 2:177-179.

Holmberg B, Lundberg P. 1985. Benzene:  Standards,  occurrence, and
exposure. Am J Ind Med  7:375-383.

Holzer G, Shanfield  H,  Zlatkis A,  et al.  1977. Collection and analysis
of  trace organic emissions from  natural sources. J Chromatog 142:755-
764.

Hopper DJ.  1978. Microbial degradation of aromatic hydrocarbons.  In:
Developments in  Biodegradation  of  Hydrocarbons.  Uatkinson RJ, ed.
London: Applied  Science, pp.  85-112.

Howard PH,  Durkin  PR.  1975.  Sources of contamination,  ambient levels and
fate of benzene  in the  environment. EPA-560/5-75-005.  Washington,  DC:
Environmental  Protection Agency (as reported in  CHEMFATE 1987).

HSDB  (Hazardous  Substances Data Bank). 1987. Offline printout of HSDB  on
benzene.  March 1987. Oak Ridge,  TN: Oak Ridge National Laboratory.

-------
144   Section 10

HSE. 1982. Toxicity Review--Benzene. United Kingdom Health and Safety
Executive. London: Her Majesty's Stationery Office (as reported in Dean
1985).

Hudak A, Ungvary G. 1978. Embryocoxic effects of benzene and its methyl
derivatives: toluene, xylene. Toxicology 11:55-63.

Hughes CS, Abshire AD. 1983. Chemical Economics Handbook.  CEH marketing
research report - benzene. Menlo Park, CA:  SRI International.

* Hunter CG. 1968. Solvents with reference  to studies on the
pharmacodynamics of benzene. Proc R Soc Med 61:913-915.

Hunter FT. 1939. Chronic exposure to benzene (benzol). II. The clinical
effects. J Ind Hyg 21:331-354 (as reported in IARC 1982).

IARC. 1974. International Agency for Research on Cancer. Monographs on
the evaluation of the carcinogenic risk of chemicals to humans. Some
antithyroid and related substances. Nitrofurans and industrial
chemicals. IARC Monogr Eval Carcinog Risk Chem Man 7:203-216.

* IARC. 1982. International Agency for Research on Cancer. Monographs on
the evaluation of the carcinogenic risk of chemicals to humans. Some
industrial chemicals and dyestuffs. IARC Monogr Eval Carcinog Risk Chem
Man 29:93-148, Suppl 4, pp. 56-57.

* Ikeda M. 1964. Enzymatic studies on benzene intoxication. J Biochem
55:231-243.

Inceman S, Tangun Y. 1969. Impaired platelet-collagen reaction in a case
of acute myeloblastic leukemia due to chronic benzene intoxication. Turk
Tip Cemiy Mecm 35:417-424 (as reported in Goldstein 1977).

Infante PF. 1978. Leukemia among workers exposed  to benzene. Tex Rep
Biol Med  37:153-161.

Infante PF, Rinsky RA, Waggoner JK, Young RJ. 1977. Leukemia in benzene
workers.  Lancet 2:76-78.

Infante PF, White MC. 1983. Benzene:  Epidemiological observations of
leukemia  by cell  type and adverse health effects  associated with low-
level exposure. Environ Health Perspect 52:75-82.

* Inoue OK, Seiji K, Kasahira M, et al. 1986. Quantitative relation of
urinary phenol  levels to breathzone benzene  concentrations: A  factory
survey.   Br J Ind Med 43:692-697.

Irons RD.  1985. Quinones  as  toxic  metabolites of  benzene. J  Toxicol
Environ Health  16:673-678.

* Irons RD, Dent  JG,  Baker  TS, Rickert  DE.  1980a.  Benzene is metabolized
and covalently  bound in  bone marrow in  situ.  Chem-Biol  Interact  30:241-
245.

-------
                                                        References   145

* Irons RD, Heck Hd'A, Moore BJ, Muirhead KA. 1979. Effects of short-
term benzene administration on bone marrow cell cycle kinetics in the
rat. Toxicol Appl Pharmacol 51:399-409.

Irons RD, Moore BJ. 1980. Effect of short term benzene administration  on
circulating lymphocyte subpopulations in the rabbit: evidence of a
selective B-lymphocyte sensitivity. Res Commun Chem Pathol Pharmacol
27(1):147-155.

Irons RD, Neptun DA. 1980. Effects of the principal hydroxy-metabolites
of benzene on microtubule polymerization. Arch Toxicol 45:297-305.

Irons RD, Neptun DA, Pfeifer RW. 1981. Inhibition of lymphocyte
transformation and microtubule assembly by quinone metabolites of
benzene: evidence for a common mechanism. J Reticuloendothel Soc
30:359-372.

* Irons RD, Wierda D, Pfeifer RW. 1980b. The immunotoxicity of benzene
and its metabolites. In: Carcinogenicity and Toxicity of Benzene.
Mehlman MA, ed. Princeton, NJ: Princeton Scientific Publishers Inc.,
pp. 37-49.

Ishimaru T, Okada H, Tomiyasu T, Tsuchimoto T, Hoshino T, Ichimaru M.
1971. Occupational factors in the epidemiology of leukemia in Hiroshima
and Nagasaki. Am J Epidemiol 93:157-165 (as reported in White et al.
1980).

* ITII. 1975. International Technical Information Institute. Toxic and
Hazardous Industrial Chemicals Safety Manual for Handling and Disposal,
with Toxicity and Hazard Data. Toyko, Japan: International Technical
Information Institute, pp. 62-64.

Jackman FA. 1975. General properties  (of benzene).  In: Benzene and Its
Industrial Derivatives. EG Hancock, ed. New York: John Wiley & Sons.
pp. 97-112.

Jacobs Engineering Co. 1978. Alternatives for Hazardous Waste Management
Industry. Pasadena, CA (as reported in Gilbert et al. 1982).

Jerina D, Daly J, Witkop B, Zaltzman-Nirenberg P, Udenfriend S.  1968.
Role of arene oxide-oxepin system  in  the metabolism of aromatic
substances. Arch Biochem Biophys 128:176-183.

Jirka AM, Bourne S. 1982. Gas-chromatographic analysis for benzene  in
blood. Clin Chem 28(7):1492-1494.

Johnson HL. 1979. Systemic absorption of benzene  -  evaluation of
experimental data and comparison of dermal and inhalation exposure.
Report submitted to Rubber Manufacturers Association, July  19  (as
reported  in Susten et al. 1985).

Jonek J, Olkowski Z, Zieleznik  B.  1965. Title not  given. Acta Histochem
20:286  (as reported  in Sandmeyer 1981).

-------
146   Section 10

Jonsson A, Berg S. 1980. Determination of 1,2-dibromomethane,
1,2-dichloroethane,  and benzene in ambient air using porous polymer
traps and gas chromatographic-mass spectrometric analysis with selected
ion monitoring. J Chromatogr 190:97-106 (as reported in IARC 1982).

JRB Associates, Inc. 1980a.- Materials Balance for Benzene, Level
I - Preliminary. Washington, DC: Environmental Protection Agency.
EPA-560/13-80-014 (as reported in IARC 1982).

JRB Associates, Inc. 1980b. Materials Balance for Benzene, level II -
McClean, VA: JRB Associates, Inc.

Kaden DA, Hites RA,  Thilly WG. 1979. Mutagenicity of soot and associated
polycyclic aromatic hydrocarbons to Salmonella cyphimurium. Cancer Res
39:4152-4159.

Kale PG, Baum JW. 1983. Genetic effects of benzene in Drosophila
melanogaster males.  Environ Mutagen 5:223-226.

Kalf GF, Post, GB, Snyder R. 1987. Solvent toxicology: Recent advances
in the toxicology of benzene, the glycol ethers, and carbon
tetrachloride. Ann Rev  Pharmacol Toxicol 27:399-427.

Kalf GF, Rushmore T, Snyder R. 1982. Benzene inhibits RNA synthesis in
mitochondria from liver and bone marrow. Chem Biol Interact 42:353-370
(also reported  in Andrews and Snyder 1986).

Karickhoff SW.  1981. Semi-empirical estimation of sorption of
hydrophobic pollutants  on natural sediments and soils. Chemosphere
10(8): 833-846.

Keller KA, Snyder CA.  1986. Mice exposed in utero to low concentrations
of benzene exhibit enduring changes in their colony forming
hematopoietic cells. Toxicology 42:171-181.

Keller KA, Snyder CA,  Dempster AM, Valle CD.  1985. Effects of benzene on
fetal hematopoiesis. Teratology  31:28A-29A.

Kenaga EE. 1980.  Predicted bioconcentration  factors and  soil sorption
coefficients of pesticides and other chemicals. Ecotoxicol Environ
Safety 4:26-38.

Khan H,  Khan MH.  1973.  Cytogenetic  studies following  chronic exposure  to
benzene.  Arch Toxikol  31:39-49.

Kimmel CA, Wilson JG.  1973.  Skeletal deviations  in  rats:  malformations
or variations?  Teratology  8:309-316.

Kimura  ET,  Ebert  DH,  Dodge  PW.  1971. Title not  given.  Toxicol Appl
Pharmacol 19:699  (as  reported in Sandmeyer 1981).

Kinoshita Y, Terada H, Saito H,  et  al.  1965.  A  case of myelogenous
 leukemia.  J  Jpn Haematol  Soc 85-96  (as reported in  Goldstein 1977).

-------
                                                        References   147

Kissling M, Speck B. 1972. Further studies on experimental benzene
induced aplastic anemia. Blut 25:97-103.

Knoll JE, Penny WH, Midgett MR. 1978. The Use of Tedlar Bags to Contain
Gaseous Benzene Samples at Source-Level Concentrations. Triangle Park,
NC: Environmental Protection Agency, Environmental Monitoring and
Support Laboratory. EPA/600/4-78/057. 49 pp.

Kociba RJ ,  Kalnins RV, Wade CE, Garfield EL, Fishbeck WA.  1976. Elevated
urinary phenol levels in beagle dogs treated with salol. Am Ind Hyg
Assoc J 37(1):183-191.

Kohli P, Brunner HE, Siegenthaler W. 1967. Erythtroleukamie nach
chronischer benzolintoxikation. Unterschung der ferro- und
erythrocytenkinetik mit radioactivem eisen und chrom. Scheiz Med
Wochenschr 97:368-373 (as reported  in Goldstein 1977).

Koizumi A,  Dobashi Y, Tachibana Y.  1979. Chromosome changes induced by
industrial chemicals. Sangyo Igaku  (Jpn J Ind Health) 21:3-10.

Koizumi A,  Dobashi Y, Tachibana Y,  Tsuda K, Katsunuma H. 1974.
Cytokinetic and cytogenetic changes  in cultured human leukocytes and
He La cells induced by benzene. Ind  Health 12:23-29.

Koljkowsky P. 1969. Indicator -tube  method for the determination of
benzene  in air. Analyst 94:918-920  (as  reported in IARC 1982).

Kowalski JD,  Shikiya JM,  Tsou  G.  1985. Ambient sampling and analysis of
halogenated hydrocarbons  and benzene. APCA  Annual Meeting,  June 16-21.
pp. 2-13.

Konasewich D, Traversy, W,  Zar H.  1978.  Status Report  on  Organic  and
Heavy Metal Contaminants  in the  Lakes  Erie, Michigan,  Huron,  and
Superior basins.  Great  Lake Water Quality Board  (as  reported  in CHEMFATE
1987 and HSDB 1987).

Kopczynski SL.  1964.  Photo-oxidation of alky Ibenzene- nitrogen dioxide
mixtures in air.  Int  J  Air  Water Pollut 8:107-120.

Korte  F, Klein  W.  1982.  Degradation of benzene  in the environment.
Ecotoxicol Environ Safety 6:311-327.

Kozioski RP.  1985.  Determination of benzene and toluene in soils  and
plant  material  by azeotropic  distillation.  Bull Environ Contain Toxicol
 Krasnoshchekova RY, Gubergrits M. 1975. Solubility of N-alkylbenzene in
 fresh and salt waters. Vodn Resur 170-173 (as reported in CHEMFATE
 1987).

-------
148   Section 10

Kraybill HF. 1977. Global distribution of carcinogenic pollutants in
water. In: Aquatic Pollutants and Biologic Effects with Emphasis on
Neoplasia. Kraybill HF, Dawe CJ, Harshbarger JC, Tardiff RG, eds.  Ann NY
Acad Sci 298:80-89.

* Kuna RA, Kapp RU. 1981. Embryotoxic/teratogenic potential of benzene
vapor in rats. Toxicol Appl Pharmacol 57:1-7.

Lakhanisky Th, Hendricks B. 1985. Induction of DNA single-strand breaks
in CHO cells in culture. In: Progress in Mutation Research 5.  Ashby J,
de Serres FJ, Draper M, Ishidate M Jr, Margolin BH,  Matter BE,  Shelby
MD, eds. Evaluation of Short-Term Tests for Carcinogens:  Report of the
International Program on Chemical Safety Collaborative Study on in vitro
Assays. Amsterdam: Elsevier, pp. 367-370.

Lange A, Smolik R, Zatonski W, Glazman H. 1973b. Leukocyte agglutinins
in workers exposed to benzene, toluene and xylene.  Int Arch Arbeitsmed
31:45-50 (as reported in Goldstein 1977).

Lange A, Smolik R, Zatonski V, Szymanska J.  1973a.  Serum immunoglobulin
levels in workers exposed to benzene, toluene and xylene.  Int Arch
Arbeitsmed 31:37-44 (as reported in Goldstein 1977).

Latriano L, Goldstein BD, Witz G. 1986. Formation of muconaldehyde,  an
open-ring metabolite of benzene, in mouse liver microsomes: An
additional pathway for toxic metabolites. Proc Natl  Acad Sci 83-8356-
8360.

* Lauwerijs R. 1979.  Human biological monitoring of  industrial
chemicals. 1. Benzene. Document EUR 6579. Health and Safety Directorate.
Commission of the European Communities, Luxemborg (as reported in van
Sittert and de Jong 1985).

Lauwerys R. 1979. Industrial health and safety. Human biological
monitoring of industrial chemicals.  I. Benzene. Luxembourg Commission of
the European Communities (as reported in IARC 1982).

Lee SD, Dourson M, Mukerjee D, Stata JF,  Kawecki J.  1983.  Assessment of
benzene health effects in ambient water.  Adv Mod Environ Toxicol 4:91-
125.

LeNoir C. 1897.  Sur un cas de purpura attribue a 1'intoxication par la
benzene. Bull Mem Soc Med Hop Paris 14:1251 In: Browning E. 1965.
Toxicity and Metabolism of Industrial Solvents. Amsterdam: Elsevier (as
reported in Aksoy 1985a).

Leo A, Hansch C, Elkins D. 1971. Partition coefficients and their uses.
Chem Rev 71(6):569.

Leong B. 1977. Experimental benzene intoxication. In: Benzene Toxicity,
a Critical Revaluation. Laskin S and Boldstein B, eds. J Toxicol Environ
Health. Suppl 2, November.

-------
                                                        References   149

Levy A.  1973. The photochemical smog reactivity of organic solvents.  In-
Solvent Theory and Practices. Adv Chem Ser 124: 70-94.

* Li G-L, Yin N, Watanabe T, et al. 1986. Benzene-specific increase in
leukocyte alkaline.phosphatase activity in rats exposed to vapors of
various organic solvents. J Toxicol Environ Health 19:581-589.

Litton Bionetics Inc., Kensington, Maryland.  Unpublished data November
1977 and December 1978 (as reported in Schwetz 1983).

Loden M. 1986. The in vitro permeability of human skin to benzene,
ethylene glycol, formaldehyde, and n-hexane.  Acta Pharmacol Toxicol
58:382-389.

Longacre S, Kocsis J, Snyder R. 1981a. Influence of strain differences
in mice on the metabolism and toxicity of benzene. Toxicol Appl
Pharmacol 60:398-409.

Longacre SL, Kocsis JJ,  Witmer CM, Lee EW, Sammett D,  Snyder R. 1981b.
Toxicological and biochemical effects of repeated administration of
benzene in mice. J Toxicol Environ Health 7:223-237.

Lorenz K, Zellner R. 1983. Kinetics of the reactions of OH radicals with
benzene, benzene-D6, and naphthalene. Ber Bunsen-Ges Physik Chem 8:629-
636.

Lowry WT, Juarez L. Petty CS, Roberts B. 1985. Studies of toxic gas
production during actual structural fires in  the Dallas area. J Forensic
Sci 30:59-72.

Ludwig H, Werthermann A. 1962. Benzene myelopathy.  Schweiz Med
Wochenschr 13:378-384 (as reported in IARC 1982).

Lurie, JB. 1952. Title not given. S Afr J Clin Sci  3:212  (as reported  in
Sandmeyer 1981).

Lutz WK, Schlatter Ch. 1977. Mechanism of the carcinogenic action of
benzene: irreversible binding  to  rat  liver DNA. Chem-Biol Interact
18:241-245.

Lyman WJ. 1982. Atmospheric  residence time.  In: Handbook  of Chemical
Property Estimation Methods. Lyman WJ, Reehl  WF,  Rosenblatt DH,  eds.  New
York: McGraw-Hill, Chapter  10.

LysyJ I, Perkins G,  Farlow JS. 1980.  Trace analysis for  aromatic
hydrocarbons  in natural  waters.  Environ  Int  4:407-416.

Lyon JP. 1976.  Mutagenicity  studies with benzene.  Dissert. Abstracts  B,
36:5537  (as  reported in  Dean 1978).

Mackay  D, Leinonen  PJ.  1975.  Rate of  evaporation of low-solubility
contaminants  from water  bodies to atmosphere. Environ Sci Technol
9:1178-1180.

-------
 150   Section  10

 Mackay D, Yeun ATK.  1983. Mass  transfer coefficient correlations for
 volatilization of organic solutes  from water. Environ Sci Technol
 17:211-217.

 Maffett PA, Doherty  TF, Monkman JL. 1956. A direct method for the
 collection and determination of micro amounts of benzene or toluene in
 air. Am Ind

 Maibach HI, Anjo DM. 1981. Percutaneous penetration of benzene and
 benzene contained in solvents in the rubber industry.  Arch Environ
 Health 36:256-260. Hyg Assoc J  17:186-188 (as reported in IARC 1982).

 Mallory TB, Gall EA, Brickley W J.  1939. Chronic exposure to benzene
 (benzol). III. The pathologic results. J Ind Hyg Toxicol 21:355-377 (as
 reported in IARC 1982).

 Maltoni C. 1983. Myths and facts in the history of benzene
 carcinogenicity. Adv Mod Environ Toxicol 4:1-15.

 Maltoni C, Ciliberti A, Carretti D. 1982a.  Experimental contributions  in
 identifying brain potential carcinogens in the petrochemical industry.
 Ann NY Acad Sci 381:216-249.

 Maltoni C, Conti B,  Cotti G. 1983. Benzene:  A multipotential carcinogen.
 Results of long-term bioassays  performed at the Bologna Institute of
 Oncology. Am J Ind Med 4:589-630.

 Maltoni C, Conti B,  Cotti G, Belpoggi F.  1985. Experimental studies on
 benzene carcinogenicity at the  Bologna Institute of Oncology:  Current
 results and ongoing  research. Am J Ind Med 7:415-446.

 Maltoni C, Conti B,  Scarnato C. 1982b. Squamous cell carcinomas of the
 oral cavity in Sprague-Dawley rats following exposure  to benzene by
 ingestion. Med Lavoro 4:441-445.

 Maltoni C, Cotti G, Valgimigli  L, Mandrioli A. 1982c.  Hepatocarcinemas
 in Sprague-Dawley rats following exposure to benzene by inhalation. Med
 Lavoro 4:446-450.

 Maltoni C, Cotti C, Valgimigli  L, Mandrioli A. 1982d.  Zymbal gland
 carcinomas in rats following exposure to benzene by inhalation.  Am J Ind
 Med 3:11-16.

Maltoni C, Scarnato C. 1977. Le prime prove sperimentali dell'azoine
 cancerogena del benzene. Gli Ospedali della Vita 4:111 (as reported in
Maltoni 1983).

Maltoni C, Scarnato C. 1979. First experimental demonstration of the
 carcinogenic effects of benzene. Long-term effects on Sprague-Dawley
 rats by oral administration. Med Lav 70:352 (as reported in Maltoni
 1983).

-------
                                                        References   151

Malysheva MV. 1980. Risk of remote consequences appearing in laboratory
animals skin-treated with benzene. Gig Tr Prof Zabol (6):51-52.

Mara SJ,  Lee SS. 1978. Assessment of Human Exposures to Atmospheric
Benzene.  EPA 450/378-031. Research Triangle Park, NC: Environmental
Protection Agency  (as reported in EPA 1985a).

Martin BE, Clark T, Burngarner J, Evans GF. 1980. Ambient Air Monitoring
for Benzene. 24-hr Integrated Sampling in Six Cities. Research Triangle
Park, NC: Environmental Protection Agency, Environmental Monitoring
Systems Laboratory. EPA 600/4-80-027.

Matheson Scientific.  (No date). Detector Tubes UNICO. Matheson
Scientific,  Elk Grove Village III, Chicago, IL (as reported in
Verschueren  1983).

Matsumoto N, lijima S, Katsunuma  H.  1975. Effect of  benzene on fetal
growth with  special reference to  the different stages of development in
mice. Cong Anom  (Senten  Ijo) 15:47-58.

McCarroll NE, Keech BH,  Piper CE.  1981a. A microsuspension adaptation of
the Bacillus subcilis "rec"  assay. Environ Mutagen  3:607-616.

McCarroll NE, Piper CE,  Keech BH.  1981b. An  E. coli  microsuspension
assay  for the detection  of DNA  damage  induced by direct-acting agents
and promutagens.  Environ Mutagen 3:429-444.

McMichael AJ, Spiritas R,  Kupper LL, Gamble  JF.  1975.  Solvent exposures
and  leukemia among rubber  workers: An epidemiologic study.  J  Occup Med
17:234  (as  reported in Snyder  1984).

Messerschmitt J.  1972. Bone-marrow aplasias during pregnancy. Nouv Rev.
Fr.  Hematol 12:15-28.

Meyne  J,  Legator MS.  1980. Sex-related differences  in cytogenetic
 effects  of benzene in the bone marrow of Swiss mice. Environ Mutagen
 2:43-50.

 Michael  LC, Erickson MD. Parks SP,  Pellizzari ED.  1980. Volatile
 environmental pollutants in biological matrices with a headspace purge
 technique.  Anal Chem 52:1836-1841.

 Michon S. 1965. Disturbances of menstruation in women working in an
 atmosphere  polluted with aromatic hydrocarbons. Pol Tig Lek  20:1648-1649
 (as reported in Barlow and  Sullivan 1982).

 * Moeschlin S, ed. 1965. Title not  given. In: Poisoning, Diagnosis and
 Treatment,  1st Amer.  ed. New York:  Grune and Stratton  (as reported in
 Sandmeyer 1978).

 Moeschlin S, Speck B. 1967. Experimental  studies on the mechanism of
 action of benzene on the  bone  marrow  (radioautographic studies  using
 3H-thymidine). Acta  Haemat 36:104-111.

-------
152   Section 10

Monson RR, Nakano EK. 1976. Mortality among rubber workers.  Am J
Epidemiol 103:284-297 (as reported in Snyder 1984).

Morimoto K. 1976. Analysis of combined effects of benzene with radiation
on chromosomes in cultured human leukocytes. Sangyo Igaku (Jpn J Ind
Health) 18:23-34.

Morimoto K. 1983. Induction of sister chromatid exchanges and cell
division delays in human lymphocytes by microsomal activation of
benzene. Cancer Res 43:1330-1334.

Morimoto K, Wolff S. 1980. Increase of sister chromatid exchanges and
perturbations of cell division kinetics in human lymphocytes by benzene
metabolites. Cancer Res 40:1189-1193.

Morimoto K, Wolff S, Koizumi A. 1983. Induction of sister-chromatid
exchanges in human lymphocytes by microsomal activation of benzene
metabolites. Mutat Res 119:355-360.

Mukhametova IM, Vozovaya MA. 1972.  Reproductive power and the incidence
of gynecological disorders in female workers exposed to the combined
effect of benzene and chlorinated hydrocarbons. Gig Tr Prof Zabol
16:6-9 (as reported in Barlow and Sullivan 1982).

Murray FJ. John JA, Rampy LW, Kuna RA, Schwetz BA. 1979. Embryotoxicity
of inhaled benzene in mice and rabbits. Am Ind Hyg Ass J 40:993-998.

Nagata T, Kageura M, Totoki K. 1978. Mass fragmentographic determination
of kerosene components in biological materials. Koenshu-lyo Masu
Kenkyukai 3:77-82, [Chem Abstr 92,  192082x] (as reported in IARC 1982).

Nahum LH, Hoff HE. 1934. The mechanism of sudden death in experimental
acute benzol poisoning. J Pharmacol Exp Ther 50:336-345.

Nakajima T, Okuyama S, Yonekura I,  Sato A. 1985. Effects of ethanol and
phenobarbital administration on the metabolism and toxicity of benzene.
Chem-Biol Interact 55:23-38.

Nawrot PS, Staples RE. 1979. Embryo-fetal toxicity and teratogenicity of
benzene and toluene in the mouse. Teratology 19-.41A.

NCI. 1977. National Cancer Institute. On Occurrence, Metabolism, and
Toxicity  Including Reported Carcinogenicity of Benzene. Summary Report.
Washington, DC (as reported in EPA 1985a and in EPA 1980).

Nesnow S, Argus M, Bergman H, et al.  1986. Chemical carcinogens: A
review and analysis of the literature of selected chemicals and the
establishment of the Gene-Tox Carcinogen Data Base. Mutat Res 185:1-195.

Newsome JR, Norman V, Keith CH.  1965. Vapor phase analysis of tobacco
smoke. Tobacco Sci 9:102-110  (as reported in EPA  1985a).

-------
                                                        References   153

NFPA (National Fire Protection Association). 1986. Fire protection guide
for hazardous materials. 9th ed, p. 49-20. Quincy, MA: National Fire
Protection Association. •

* NIOSH (National Institute for Occupational Safety and Health).  1974
Criteria for a Recommended Standard ... Occupational Exposure to
Benzene. Washington, DC: National Institute for Occupational Safety and
Health, Department of Health, Education, and Welfare. NIOSH-74-137.

NIOSH (National Institute for Occupation Safety and Health). 1977a.
Revised Recommendation for an Occupational Exposure Standard for
Benzene. Cincinnati, OH: National Institute for Occupational Safety and
Health.

NIOSH (National Institute of Occupational Safety and Health).  1977b.
NIOSH Manual of Analytical Methods, Vol. 3, 2nd ed. Cincinnati, OH:
NIOSH,  Department of Health, Education, and Welfare, pp. S-311-1-8.

NIOSH (National Institute for Occupational Health and Safety).  1984.
NIOSH Manual of Analytical Methods, Vol. 1. 3rd ed., Eller PM,  ed.
Cincinnati, OH: NIOSH, Department of Health and Human Services,
pp. 8305-1-8305-4.

NIOSH (National Institute for Occupational Safety and Health).  1986.
NIOSH recommendations for occupational safety and health standards,
September 1986. Morbid Mortal Week Report 35 (IS), 33 pp.

Nissen NI,  Soeborg OA. 1953. Erythromyelosis (morbus di Guglielmo).
Review and report of a case in a benzene (benzol) worker. Acta Med Scand
145:56-71 (as reported in IARC 1982).

Nojima K,  Fukaya K, Fukui S, Kanno S.  1975. Studies on photochemistry of
aromatic hydrocarbons II.  Chemosphere  2:77-82.

* Nomiyama K, Nomiyama H.  1974a. Respiratory retention, uptake and
excretion of organic solvents in man.  Benzene, toluene, n-hexane,
trichloroethylene, acetone, ethyl acetate and ethyl alcohol. Int Arch
Arbeitsmed 32:75-83 (as reported in IARC 1982).

* Nomiyama K, Nomiyama H.  1974b. Respiratory elimination of organic
solvents in man. Benzene,  toluene, n-hexane, trichloroethylene, acetone,
ethyl acetate and ethyl alcohol. Int Arch Arbeitsmed  32:85-91  (as
reported in IARC 1982).

NRC (National Research Council). 1980a. Drinking water and health. Vol.
3, Washington, DC: National Academy Press, pp. 80-86, 261-262.

NRC (National Research Council). 1980b. The Alky Benzenes. Committee on
Alkyl Benzene Derivatives. Washington, DC. National Academy  Press (as
reported in EPA 1983).

-------
  154    Section  10

  NTP  (National  Toxicology Program).  1985.  Fourth Annual Report  on
  Carcinogens. Summary. Research  Triangle Park, NC: NTP, Department of
  Health and Human  Services,-  Public Health  Service.

  * NTP  (National Toxicology  Program). 1986. Toxicology and Carcinogenesis
  Studies of Benzene  (CAS  No.  71-43-2) in F344/N Rats and B6C3F1 Mice
  (Gavage Studies). Research  Triagle  Park. NC: National Toxicology
  Program.                                                      67

  Nutmagul V, Cronn DR. 1985.  Determination of selected atmospheric
  aromatic hydrocarbons at  remote continental and oceanic locations using
  pnotoionization/flame ionization detection. J Atmos Chem 24(4) :415-433

  Oberly TJ .  Bewsey BJ , Probst GS . 1984.  An evaluation of the L5178Y TK+/-
                forward mutati
-------
                                                        References   155

* Pagnotto LD, Elkins HB, Brugsch HG, Walkley EJ .  1961. Industrial
benzene exposure from petrochemical naphtha - I. Rubber coating
industry. Am Ind Hyg Assoc J 22:417-421.

Painter RB, Howard R. 1982. The HeLa-DNA synthesis inhibition test as a
rapid screen for mutagenic carcinogens. Mutat Res 92:427-437.

Parke DV, Williams, RT.  1953a. Studies in detoxication. 49. The
metabolism of benzene containing [ 14C1] benzene .  Biochem J 54:231-238.

Pate CT. Atkinson R, Pitts JN, Jr. 1976. The gas phase reaction
of 03 with a series of aromatic hydrocarbons. J Environ Sci Health
Pavlenko GI, Domshlak MG, Chirkova EM, Katosova LD. 1979. Results of
studying the mutagenic activity of benzene. Toksikol Nov Prom Khim
Veshchestv 15:30-33.

Pellack-Walker P, Blumer JL.  1986. DNA damage  in L5178YS cells following
exposure to benzene metabolites. Mol  Pharmacol 30:42-47.

Pereira RW, Hughes BA. 1980.  Determination of  selected volatile organic
priority pollutants in water  by computerized gas chromatography-
quadrupole mass  spectrometry .  J Am Water Works Assoc 72:220-230 (as
reported in IARC 1982).

Pfeifer R, Irons R. 1981.  Inhibition  of  lectin- stimulated  lymphocyte
agglutination and mitogenesis by hydroquinone : reactivity  with
intracellular sulfhydryl  groups. Exp  Mol Pathol  35:189-198.

Philip P, Krogh  Jensen M.  1970.  Benzene  induced  chromosome abnormalities
in rat bone marrow  cells.  Acta Pathol Microbiol  Scand  Sect A 78:489-490.

Pinigina IA, Mal'tseva NM.  1978. Determination of  benzene  in blood.  Gig
Sanit 12:67-71  [Chem  Abstr 90, 97238c]  (as reported in IARC 1982).

Pitts JN, Jr, Darnall K,  Carter WPL,  Winer AM, Atkinson R. 1979.
Mechanisms  of photochemical reactions in urban air. Research Triangle
Park, NC:  Environmental  Protection Agency. EPA- 600/3 -79 -110.
p. 62-63.

 Pollini  G,  Biscaldi GP,  Robustelli Delia Cuna G. 1969. Chromosome
 changes  in lymphocytes five years after benzene haemopathy. Med Lav
 60:743-758.

 Post GB,  Snyder R,  Kalf GF.  1985.  Inhibition of RNA synthesis and
 interleukin-2 production in  lymphocytes in vitro by benzene and its
 metabolites,  hydroquinone and p-benzoquinone. Toxicol Lett 29:161-167.

-------
156   Section 10

Probst GS, Hill LE. 1985. Tests for the induction of DNA-repair
synthesis in primary cultures of adult rat hepatocytes.  In: Progress in
Mutation Research 5. Ashby J, de Serres FJ, Draper M, Ishidate M Jr,
Margolin BH, Matter BE, Shelby MD, eds. Evaluation of Short-Term Tests
for Carcinogens: Report of the International Program on Chemical Safety
Collaborative Study on*in'vitro Assays. Amsterdam: Elsevier,  pp. 381-
386.

Probst GS, McMahon RE. Hill LE, Thompson CZ, Epp JK,  Neal SB.  1981.
Chemically-induced unscheduled DNA synthesis in primary rat hepacocyte
cultures: A comparison with bacterial mutagenicity using 218  compounds.
Environ Mutagen 3:11-32.

Pushkina NN, Gofmekler VA, Klevtsova GN.  1968.  Changes in content of
ascorbic acid and nucleic acids produced by benzene and formaldehyde.
Bull Exp Biol Med (USSR) 66:868-870 (translated from Byull Eksp Biol Med
66:51-53).

Reineke FJ, Baechmann. 1985. Gas chromatographic determination of
2-carbon to 8-carbon hydrocarbons and halocarbons in ambient air by
simultaneous use of 3-detectors. J Chromatogr 323(2):323-330.

Renova NV. 1962. Concerning auto-immunity shifts in chronic occupational
benzol poisoning. Gig Tr Prof Zabol 7:38-42 (as reported in Goldstein
1977).

* Rickert DE, Baker TS, Bus JS, et al. 1979. Benzene disposition in the
rat after exposure by inhalation. Toxicol Appl Pharmacol 49:417-423.

Rinsky, RA, Alexander B, Smith MD, et al. 1987. Benzene and leukemia: An
epidemiological risk assessment. N Eng J Med 316:1044-1050.

* Rinsky RA, Young RJ, Smith AB. 1981. Leukemia in benzene workers.  Am J
Ind Med 2:217-245.

Roberts JM, Fehsenfeld FC, Liu, SC, et al. 1984. Measurements of
aromatic hydrocarbon ratios and nitrogen oxide concentrations in the
rural troposphere: Observation of air mass photochemical aging and
nitrogen oxides removal. Atmos Environ 18(11):2421-2432.

Roberts JM, Hutte RS, Fehsenfeld FC, Albritton DL, Slevers RE. 1985.
Measurements of anthropogenic hydrocarbon concentration ratios in the
rural troposphere: discrimination between background and urban sources.
Atmos Environ 19(11):1945-1950.

Rogers RD, MeFarlane JC, Cross AJ. 1980. Adsorption and desorption of
benzene in two soils and monmorillonite clay. Environ Sci Techno1
14:457-460.

* Rosenthal GJ, Snyder CA. 1985. Modulation of the immune response to
Lister La caonocytogenes by benzene inhalation. Toxicol Appl Pharmacol
80:502-510.

-------
                                                         References    157

 Rosenthal GJ,  Snyder CA.  1986.  Altered T-cell responses  In C57BL/6J  mice
 following sub-chronic benzene Inhalation.  lexicologist 6(1):68.

 Roth L,  Kinu IV,  Turcanu P,  Moise G.  1972.  Cytologic  and immunochemical
 features of benzene induced reticuloses.  Timisoara Med 17:29-38  (as
 reported in Goldstein 1977).

 Roth L,  Turcanu P,  Dinu I,  Moise  G.  1973.  Monocytosis  in those who work
 with benzene and chronic  benzene  poisoning.  Folia  Haematol  100:213-224
 (as  reported in Goldstein 1977).

 Roush GJ,  Ott  MG.  1977. A study of benzene  exposure versus urinary
 phenol levels.  Am Ind Hyg Assoc J 38:67-75.

 Rozen MG,  Snyder  CA.  1985.  Protracted exposure  of  C57B1/6 mice to 300
 ppm  benzene  depresses B-  and  T-lymphocyte  numbers  and  mitogen responses.
 Evidence for thymic and bone  marrow proliferation  in response to the
 exposures. Toxicology 37(1-2):13-26.

 * Rozen  MG,  Snyder  CA,  Albert RE.  1984. Depressions in B- and T-
 lymphocyte mitogen-induced blastogenesis  in  mice exposed to low
 concentrations  of benzene. Toxicol Lett 20:343-349.

 Rozman C, Woessner  S,  Saez-Serrania J. 1968.  Acute  erythromyelosis after
 benzene  poisoning.  Acta Haematol  40;234-237  (as reported in IARC 1982).

 RTECS. 1987. Registry of  Toxic  Effects of Chemical  Substances. Online
 file,  current  file  83/8303  (also  reported  in HSDB  1987).

 Ruedemann R, Deichmann WB. 1953.  Blood phenol level after topical
 application  of  phenol-containing  preparations. JAMA 152:506-509.

 Rushmore T,  Snyder  R,  Kalf G. 1984. Covalent binding of  benzene and  its
 metabolites  to  DNA  in rabbit bone marrow mitochondria  in vitro. Chem-
 Biol  Interact 49:133-154.

 Rushton  L, Alderson M.  1981. A  case-control  study  to investigate the
 association between exosure to  benzene and deaths  from leukaemia in oil
 refinery workers. Br J  Cancer 43:77-84.

 Sabourin P, Chen B, Henderson R,  Lucier G, Birnbaum L. 1986. Effect of
 dose on absorption  and excretion  of ^C benzene administered orally or
by inhalation.  The  Toxicologist 6:163.

 SammetC D, Lee  EV,  Kocsis JJ, Snyder  R. 1979. Partial  hepatectomy
 reduces  the metabolism and toxicity of benzene. J Toxicol Environ Health
 5:785-792.

* Sandmeyer EE. 1981. Aromatic  hydrocarbons.  In: Patty's Industrial
Hygiene and Toxicology, Vol. 2, 3rd rev. ed., Clayton  GD, Clayton, FE,
eds.  New York:  Interscience Publishers, pp.  3253-3283.

-------
 158   Section  10

 Sato A, Fuglwara Y, Nakajima, T.  1974. Solubility of benzene, toluene
 and m-xylene In various body fluids and tissues of rabbits. Sangyo Izaku
 16:30.

 Sato A, Nakajima T. 1985. Enhanced metabolism of volatile hydrocarbons
 in rat  liver following food deprivation, restricted carbohydrate intake,
 and administration of ethanol, phenobarbital, polychlorinated biphenyl
 and 3-methylcholanthrene: A comparative study. Xenobiotica 15:67-75.

 Sato A, Nakajima T. 1979. Partition coefficients of some aromatic
 hydrocarbons and ketones  in water, blood,  and oil.  Toxicol Appl
 Pharmacol 48:49 (as reported in HSDB 1987).

 Sato A, Nakajima T, Fujiwara Y, et al. 1975. Kinetic studies on sex
 difference in  susceptibity to chronic benzene intoxication - with
 special reference to body fat content. Br J Ind Med 32:321-328.

 Sato A, Nakajima T, Koyama Y. 1980. Effects of chronic ethanol
 consumption on hepatic metabolism of aromatic and chlorinated
 hydrocarbons in rats. Br J Ind Med 37:382-386.

 Sato A, Nakajima T, Koyama Y. 1981. Dose-related effects of a single
 dose of ethanol on the metabolism in rat liver of some aromatic and
 chlorinated hydrocarbons. Toxicol Appl Pharmacol 60:8-15.

 Sauer TC, Jr.  1981. Volatile organic compounds in open ocean and coastal
 surface waters. Org Geochem 3:91-101.

 Savchenko MF.  1967. Title not given. Gig Sanit 32:349 (as reported in
 Sandmeyer 1981).

 Sawhney BL, Kozloski RP.  1984.  Organic pollutants in leachates from
 landfill sites. J Environ Quality 13(3):349-352.

 Saxton J, Narkus-Kramer M. 1975. EPA findings of solid waste from
 industrial chemicals. Chem Eng pp. 107-112.

* Schrenk HH, Yanc WP, Pearce SJ, et al.  1941. Absorption, distribution
and elimination of benzene by body tissues and fluids of dogs exposed to
benzene vapor.  J Ind Hyg Toxicol 23:20-34.

Schultz B, Kjeldsen P. 1986. Screening of organic matter in leachates
 from sanitary  landfills using gas chromatography combined with mass
spectrometry. Water Res 20(8):965-970.

Schwetz BA. 1983.  A review of the developmental toxicity of benzene.
 1968.  Adv Mod Environ Toxicol 4:17-21.

Seidenberg JM,  Anderson DC,  Becker RA. 1986. Validation of an in vivo
developmental toxicity screen in the mouse. Teratog Carcinog Mutagen
5:361-374.

-------
                                                        References   159

Seixas GM, Andon BM, Hollingshead PG, Thilly WG. 1982. The aza-arenes as
mutagens for Salmonella cyphimuriwn. Mutat Res 102:201-212.

Sellyel M, Kelemen E. 1971. Chromosome study In a case of granulocytic
leukemia with "pelgerisation" seven years after benzene pancytopenia.
Eur J Cancer 7:83-8>5. -

Setzkorn EA, Huddleston RL. 1965. Ultraviolet spectroscopic analysis for
following the biodegradation of hydrotropes. J Amer Oil Chem Soc
42:1081-1084.

Shackelford VM, Cline DM. 1983. An evaluation of automated spectrum
matching for survey identification of wastewater components by gas
chromatography-mass spectrometry. Anal Chim Acta 146:15-27.

Shackelford WM, Keith LH. 1976. Frequency of organic compounds
identified in water. Athens, Georgia: Environmental Research Laboratory,
Environmental Protection Agency. EPA-600/4-76-062.

Shahin MM, Fournier F. 1978. Suppression of mutation induction and
failure to detect mutagenic activity with athabasca tar sand fractions.
Mutat Res 58:29-34.

Schairer LA, Van't Hof J, Hayes CG, Burton RM, de Serres FJ. 1979.
Measurement of biological activity of ambient air mixtures using a
mobile laboratory for  in situ exposures: Preliminary results from the
Tradescantia plant test system.  Environ Sci Res 15:419-440.

Sharma RK, Jacobsen-Kram D, Lemmon M, Bakke J, Galperin I, Blazak WF.
1985. Sister-chromatid exchange  and cell replication kinetics in fetal
and maternal cells after treatment with chemical  teratogens. Mutat Res
158:217-231.

Sherwood RJ. 1972. Benzene:  the  interpretation  of monitoring results.
Ann Occup Hyg  15:409-421.

Sherwood RJ, Carter  FUG. 1970. The  measurement  of occupational  exposure
to benzene vapour. Ann Occup Hyg 13:125-146  (as reported  in IARC  1974).

Shimizu M, Yasui Y,  Matsumoto  N.  1983.  Structural specificity of
aromatic  compounds with  special  reference  to  mutagenic activity in
Salmonella  typhimuriuo--a  series of chloro-  or  fluoro-nitrobenzene
derivatives. Mutat  Res 116:217-238.

Sina  JF,  Bean  CL, Dysart GR,  Taylor VI,  Bradley MO.  1983.  Evaluation of
the alkaline elution/rat hepatocyte assay  as  a  predictor  of
carcinogenic/mutagenic potential.  Mutat Res 113:357-391.

Singh GB,  Salas  LJ,  Cantrell BK, Redmond RM.  1985.  Distribution of
aromatic  hydrocarbons in ambient air.  Atmos Environ 19(11):1911-1919.

-------
 160   Section 10

 Singh HB, Salas LJ,  Stiles RE. 1982. Distribution of selected gaseous
 organic mutagens and suspect carcinogens in ambient air.  Environ Sci
 Technol 16(12):872-880.

 Siou G, Conan L, El  Haitem M. 1981. Evaluation of the clastogenic action
 of benzene by oral administration with 2 cytogenetic techniques in mouse
 and Chinese hamster.  Mutat Res 90:273-278.

 Sittig M. 1981.  Handbook of toxic and hazardous chemicals   Park Ridze
 NJ:  Noyes Publications, pp. 75-78.                                    '

 Smart RC, Zannoni VG.  1985. Effect of ascorbate on covalent binding of
 benzene and metabolites to isolated tissue  preparations. Toxicol Apol
 Pharmacol 77:334-343.                                              *V

 Smillie RD,  Sakuma T,  Duholke WK.  1978.  Low molecular weight aromatic
 hydrocarbons in  drinking water.  J  Environ Sci  Health A13(2):  187-197.

 Smith JH,  Bomberger DC, Jr, Haynes  DL.  1980. Prediction of  the
 volatilization rates of high-volatility chemicals  from natural  water
 bodies.  Environ  Sci Technol 14:1332-1337.

 Smith RV,  Rosazza JP.  1974. Microbial  models of mammalian metabolism.
 Aromatic hydroxylation. Arch Biochem Biophys 161:551-558.

 Smolik R,  Grzybek-Hryncewicz K,  Lange  A,  Zatonski W.  1973.  Serum
 complement level in workers exposed to benzene,  toluene and xylene.  Int
 Arch  Arbeitsmed  31:243-247  (as reported in  Goldstein 1977).

 Smyth HF,  Carpenter CP,  Weil CS, Pozzani  UC, Striegel  JA. 1962.  Range-
 finding toxicity data:  List VI. J Ind  Hyg Assoc  23(1):95-107.

 Snyder CA.  1987.  Benzene.  In: Ethyl  Browning's  Toxicity and Metabolism
 of Industrial  Solvents,  2nd ed., Vol.  1:  Hydrocarbons. Snyder R, ed.

 Snyder CA, Baarson K, Goldstein, GD, Albert R.   1981a.  Ingestion  of
 ethanol  increases  the hematotoxicity of  inhaled benzene in C57BL mice.
 Bull  Environ Contam Toxicol 27:175-180.

 Snyder CA, Goldstein BD.  Sellakumar A.  1978. Hematotoxicity^of  inhaled
benzene  to Sprague-Dawley rats and AKR mice at  300 ppm. J Toxicol
 Environ  Health 4:605-618.

Snyder CA, Goldstein BD,  Sellakumar AR, Albert RE. 1984.  Evidence for
hematotoxicity and tumorigenesis in rats exposed to  100 ppm benzene. Am
J Ind Ned  5:429-434.

Snyder CA, Goldstein BD,  Sellakumar AR, Bromberg I, Laskin S, Albert RE.
1980. The  inhalation toxicology of benzene:  Incidence of hematopoietic
neoplasms and hematotoxicity  in AKR/J and C57BL/6J mice.  Toxicol Appl
Phann 54:323-331.

-------
                                                         References    161

 Snyder CA,  Green JD,  LoBue J,  Goldstein BD,  Valle CD,  Albert RE.  1981b.
 Protracted  benzene exposure causes a proliferation of  myeloblasts and/or
 promyelocytes in CD-I mice. Bull Environ Contain Toxicol  27:17-22.

 Snyder CA,  Erlichman M,  Goldstein BD,  Laskin S.  1977.  An extraction
 method for  determination of benzene in tissue by gas chromatography.  Am
 Ind Hyg Assoc J  38:272-276 (as reported in C.A.  Snyder 1987).

 Snyder CA,  Erlichman MN,  Laskin S,  Goldstein BD,  Albert  RE.  1981c. The
 pharmacokinetics of repetitive benzene exposures at 300  and  100 ppm in
 AKR mice and  Sprague-Dawley rats.  Toxicol  Appl Pharmacol 57:164-171.

 Snyder CA,  Laskin S,  Goldstein B.  1975.  An extractive  method for
 determination of benzene  in blood by gas chromatography.  Am  Ind Hyg
 Assoc  J  36(11):833-836.

 Snyder R. 1984.  The benzene problem in historical  perspective. Fundam
 Appl Toxicol  4:  692-699.

 Snyder R, Kocsis JJ.  1975.  Current  concepts  of chronic benzene toxicity.
 CRC Crit Rev  Toxicol  3:265-88  (as  reported in Andrews  and Snyder 1986).

 Snyder R, Lee  EW,  Kocsis  JJ. 1978.  Binding of labeled  metabolites to
 mouse  liver and  bone  marrow. Res  Commun Chem Pathol Pharmacol
 20:191-194.

 Snyder R, Lee  EW,  Kocsis  JJ, Vitmer CM.  1977.  Bone  marrow depressant and
 leukemogenic  actions  of benzene.  Life  Sciences 21:1709-1722.

 Snyder R, Longacre  SL, Vitmer  CM, Kocsis JJ.  1981.  Biochemical
 toxicology of benzene. In:  Reviews  in  Biochemical Toxicology 3. Hodgson
 E, Bend  JR, Philpot RM, eds. New York:  Elsevier/North  Holland,
 pp. 123-153.

 Sporstoel S, Urdal  K, Drangsholt H,  Gjoes  N.  1985.  Description of a
 method for automated  determination  of  organic  pollutants  in  water. Int J
 Environ Anal Chem 21:129-138.

 Srbova J, Teisinger J, Skramovsky S. 1950. Absorption  and elimination of
 inhaled benzene  in  man. Arch Ind Hyg Occup Med 2:1-8.

 Stetter JR,  Jurs  PC,  Rose SL.  1986.  Detection  of hazardous gases and
vapors:  Pattern  recognition analysis of data from an electochemical
 sensor array.  Anal  Chem 58(4):860-866.

 Stoner RD, Drew RT, Bernstein  DM. 1981.  Benzene  inhalation effect upon
 tetanus antitoxin responses and leukemogenesis in mice.  In:  Coal
Conversion and the  Environment. Mahlum DD, Gray RH, Felix WD, eds. Oak
Ridge,  TN: Technical  Information Center, U.S.  Department  of  Energy,  pp.
445-461.

Strong LC. 1977.   Genetic  and environmental interactions.   Cancer
40:1861-1866.

-------
162   Section 10

Stuart JD, Luce HD, Smith SR, Black RF. 1984.  Detection and Effects of
Aperiodic Leachate Discharges from Landfills.  Technical Report.  Storrs,
CT: Institute of Water Resources, University of Connecticut.

Stuart JD, Luce HD, Smith SR, et al. 1985. Institute of Water Resources,
University of Connecticut. Detection and Effects of Aperiodic Leachate
Discharges from Landfills. U.S. Department of the Interior. Technical
Completion Report G832-04.

Styles JA, Richardson CR. 1984. Cytogenetic effects of benzene:
dosimetric studies on rats exposed to benzene vapor. Mutat Res
135:203-209.

Susten A, Dames B, Burg J, Niemeier R. 1985. Percutaneous penetration of
benzene  in hairless mice: An estimate of dermal absorption during tire-
building operations. Am J Ind Med 7:323-335.

Swenberg JA, Petzold GL, Harbach PR. 1976. In vitro DNA damage/alkaline
elution  assay  for  predicting carcinogenic potential. Biochem Biophys Res
Commun 72:732-738.

Tabak HH, Quave SA, Mashni CI,  Barth EF.  1981. Biodegradability studies
with  organic priority pollutant compounds. J Water  Pollut  Control Fed
53:1503-1518.

Tabershaw JR,  Lamm SH.  1977.  Benzene and  leukemia.  Lancet  2:867-868  (as
reported in  Cole  and Merletti  1980).

Tanooka  H.  1977.  Development and applications  of Bacillus  subcilis  test
systems  for  mutagens,  involving DNA-repair  deficiency  and  suppressible
auxotrophic  mutations.  Mutat Res 42:19-32.

Tareef EM,  Kontchalovskaya NM.  Zorina LA.  1963.  Benzene  leukemias.  Acta
Unio Int Cancrum 19:751-755 (as reported  in '.ARC  1982).

Tatken RL,  Lewis RJ,  Sr.  1983.  Registry of Toxic  Effects of Chemical
 Substances.  1981-1982  edition.  Cincinnati,  OH: National  Institute for
 Occupational Safety and Health.

 Tatrai E, Rodics K, Ungvary GY. 1980b. Embryotoxic effects~of
 simultaneously applied exposure of benzene and toluene.  Folia Morphol
 (Prague) 28:286-289.

 Tatrai E, Ungvary GY,  Hudak A, Rodics K,  Loerincz M, Barcza GY. 1980a.
 Concentration dependence of the embryotoxic effects of benzene
 inhalation in CFY rats. J Hyg  Epidemiol Microbiol Immunol 24:363-371.

 Tauber J. 1970. Instant benzol death. J Occup Med 12:91-92.

 Teisinger J, Bergerova-Fiserova V, Kudrna J.  1952. The metabolism of
 benzene  in man. Pracov Lek 4:175-188.

-------
                                                        References   163

Thibodeaux LJ.  1981. Estimating the air emissions of chemicals from
hazardous waste landfills. J Hazard Mat 4:235-244.

* Thienes H, Haley TJ.  1972. Title not given. In: Clinical Toxicology.
Philadelphia, PA: Lea & Fegiger (as reported in Sandmeyer 1981).
                   »   r
Thomas RG. 1982. Volatilization from water. In: Handbook of Chemical
Property Estimation Methods. Lyman WJ, Reehl WF, Rosenblatt DH,  eds. New
York: McGraw-Hill, Chapter 16.

Thorpe JJ. 1974. Epidemiologic survey of leukemia in persons potentially
exposed to benzene. J Occup Med 18:375-382 (as reported in Infante and
White 1983).

Tice RR, Costa DL, Drew RT. 1980. Cytogenetic effects of inhaled benzene
in murine bone marrow:  Induction of sister chromatid exchanges,
chromosomal aberrations and cellular proliferation inhibition in DBA/2
mice. Proc Natl Acad Sci USA 77:2148-2152.

Tice RR, Vogt TF, Costa DL. 1982. Cytogenetic effects of inhaled benzene
in murine bone marrow,  In: Genotoxic Effects of Airborne Agents.  Environ
Sci Res 25:257-275.

* Toft K, Olofsson T, Tunek A, Berlin M. 1982. Toxic effects on mouse
bone marrow caused by inhalation of benzene. Arch Toxicol 51:295-302.

Topham JC. 1980. Do induced sperm-head abnormalities in mice
specifically identify mammalian mutagens rather  than carcinogens? Mutat
Res 74:379-387.

Torres A, Giralt M, Raichs A.  1970. Coexistencia de antecedents
benzolicas cronicos 4 plasmocitoma multiple  presentation de dos coses.
Sangre 15:275-279  (as reported in Aksoy 1985a).

Tough IM, Court  Brown WM.  1965. Chromosome aberrations and exposure to
ambient benzene. Lancet 1:684.

Tsai SP,  Wen CP, Weiss NS, Wong 0, McClellan WA,  Gibson RL. 1983.
Retrospective mortality and medical  surveillance studies  of workers in
benzene areas of refineries.  J Occup  Med  25:685-692.   <•

Tunek A,  Olofsson T, Berlin M. 1981.  Toxic effects  of benzene and
benzene metabolites on granulopoietic stem cells and bone marrow
cellularity in  mice. Toxicol  Appl  Pharmacol  59:149-156.

Tzanck A, Dreyfus  A, Jais M.  1937.  Hemopathie  post-benzolique et
leucoblastose medullaire. Sangre  11:550-557  (as reported in Goldstein
1977).

Umana M,  Warner M, Sheldon LS. 1985.  Methods for Sampling and Analysis
of Breath.  Environmental  Protection Agency.  EPA 600/8-85/019. NTIS
PB85-243277.

-------
164   Section 10

Unger HT, Claff RE. 1985. Evaluation of percent removal variability for
priority pollutants in POTWS. Proc 40th Ind Waste Conf, Purdue
University, Vest Lafayette, Indiana, May 14-16, 1985.

Ungvary G. 1985. The possible contribution of industrial chemicals
(organic solvents) to the incidence of congenital defects caused by
teratogenic drugs and consumer goods: An experimental  study.  Prog Clin
Biol Res 1638:295-300.

* Ungvary G, Tatrai E. 1985. On the embryotoxic effects of benzene and
its alkyl derivatives in mice, rats and rabbits.  Arch  Toxicol (Suppl
8):425-430.

Valle-Paul C, Snyder CA. 1986. Effect of benzene exposure on bone marrow
precursor cells of splenectomized mice. Toxicologist 6(1):285.

Van den Berghe H, Louvagie A, Broeckaert-Van Orshoven  A, David G,
Vervilghen R. 1979. Chromosome analysis in two unusual malignant blood
disorders presumably induced by benzene. Blood 53:558-566.

Van Duuren BL, Sivak A, Goldschmidt BM, Katz C, Melchionne S. 1969.
Carcinogenicity of halo-ethers. J Natl Cancer Inst 43:481-486.

van Raalte HGS, Grasso P. 1982. Hematological, myelotoxic, clastogenic,
carcinogenic, and leukemogenic effects of benzene. Regul Toxicol
Pharmacol 2:153-176.

* Van Sittert NJ, de Jong G. 1985. Biomonitoring of exposure to
potential mutagens and carcinogens in industrial populations. Fd Chem
Toxic 23(1):23-31.

Vara P, Kinnunen 0. 1946. Benzene toxicity as a gynecologic problem.
Acta Obstet Gynecol Scand 26:433-452.

Versar, Inc. 1979. Water-related Environmental Fate of 129 Priority
Pollutants..VI. Phenols, Cresols, and Monocyclic Aromatics. Draft
report. Washington, DC: Office of Water Planning and Standards,
Environmental Protection Agency. Contract No. 68-01-3852, EPA-44014-79-
029E.

Verschueren K. 1983. Handbook of Environmental Data on Organic
Chemicals. 2nd edition, pp. 236-242. New York: Van Nostrand Reinhold Co.

Vianna NJ, Polan A. 1979. Lymphomas and occupational benzene exposure.
Lancet 1:1394-1395 (as reported in Aksoy 1985).

View Data Base. 1989. Agency for Toxic Substances and Disease Registry
(ATSDR), Atlanta, GA: Office of External Affairs, Exposure and Disease
Registry Branch, February 1989.

Vigliani EC. 1976. Leukemia associated with benzene exposure. Ann NY
Acad Sci 271:143-151  (as reported in Kalf et al. 1987, C. Snyder 1987).

-------
                                                        References   165

Vigliani EC, Fornl A. 1976. Benzene and leukemia. Environ Res 11:122-127
(as reported in Infante and White 1983, and in C. Snyder 1987).

Vigliani EC, Saita G. 1964. Benzene and leukemia. New Engl J Med
271(17):-872-876.

Viraraghavan T. Hashem S. 1986. Trace organics in septic tank effluent.
Water Air Soil Pollut 28:299-308.

Von Oettingen WF. 1940. Title not given. Public Health Bulletin No.  255
Washington, DC: U.S. Public Health Service (as reported in Sandmeyer
1981).

Wallace LA. 1986. The Total Exposure Assessment Methodolgy (TEAM) Study.
Part I: Summary and Analysis. Part II: Protocols for Environmental and
Human Sampling and Analysis. Final report to the Environmental
Protection Agency.

Wallace LA, Pellizzari Ed, Hartwell TD, Sparacino CM. Sheldon LS, Zelon
H. 1985. Personal exposures, indoor-outdoor relationships, and breath
levels of toxic air pollutants measured for 355 persons in New Jersey.
Atmos Environ 19:1651-1661.

Wallace LA, Pellizzari ED. 1986. Personal air exposures and breath
concentrations of benzene and other volatile hydrocarbons for smokers
and nonsmokers. Toxicol  Lett 35:113-116.

Wallace LA, Pellizzari E, Leader B, Zelon H, Sheldon L. 1987. Emissions
of volatile organic compounds  from building materials and consumer
products. Atmos  Environ  21(2):385-393.

* Ward CO, Kuna  RA,  Snyder NK, Alsaker RD, Coate WB, Craig  PH.  1985.
Subchronic  inhalation  toxicity of benzene  in rats  and mice.  Am  J Ind  Med
7:457-473.

Weast RC,  et al.  1979. CRC Handbook of Chemistry and Physics.  60th  ed.,
p. C-146  (as reported  in HSDB  1987).

Weast RC,  Astle  HJ,  Beyer WH,  eds.  1985.  Handbook  of Chemistry and
Physics.  66th  ed.,  Boca  Raton,  FL:  CRC Press,  Inc.  p.  C-105.

Weiss G.  1980. Hazardous Chemicals  Data Book.  Park Ridge,  NJ:  Noyes Data
Corporation, p.  143.

* Wester  RC, Maibach HI, Gruenke LD.  Craig JC.  1986. Benzene levels in
ambient air and breath of smokers and nonsmokers in urban and pristine
environments.  J  Toxicol  Environ health 18:567-573.

White MC,  Infante PF,  Walker B.  1980. Occupational exposure to benzene:
A review  of the carcinogenic and related health effects following the
United States  Supreme Court decision. Am J Ind Med 1:233-244.

-------
166   Section 10

WHO (World Health Organization). 1984. Guidelines for drinking water
quality. Vol. 2. Health criteria and other supporting information.
Geneva, Switzerland: World Health Organization.

Wierda D, Greenlee WF(> Irons RD. 1980. Immunetoxicity of benzene
metabolites  in C57BL/6 mice. Pharmacologist 22:207A (as reported in
Wierda et al. 1981).

Wierda D. Irons RD. 1981. Reduction or progenitor B-lymphocytes in mice
after hydroquinone and catechol administration. Fed Proc 40:675A (as
reported in Wierda et al. 1981).

Wierda D, Irons R. 1982. Hydroquinone and catechol reduce the frequency
of progenitor B lymphocytes in mouse spleen and bone marrow.
Immunopharmacology 4:41-54.

Wierda D, Irons RD, Greenlee WF. 1981. Immunotoxicity in C57BL/6 mice
exposed to benzene and aroclor 12S4. Toxicol Appl Pharmacol 60:410-417.

Wilcosky TC, Checkoway H. Marshall EC. Tyroler HA. 1984. Cancer
mortality and solvent exposures in the rubber industry.  Am J Ind Hyg
Assoc 45:809-811.

Williams CM, Tong C, Ved Brat S. 1985. Tests with the rat hepatocyte
primary culture/DNA-repair test. In: Progress in Mutation Research 5.
Ashby J, de Serres FJ,  Draper M, Ishidate M Jr, Margolin BH, Matter BE,
Shelby MD, eds. Evaluation of Short-Term Tests for Carcinogens: Report
of the International Program on Chemical Safety Collaborative Study on
in vitro Assays. Amsterdam: Elsevier, pp. 341-345.

Wilson BH, Smith GB, Rees JF. 1986. Biotransformation of selected
alkylbenzenes and halogenated hydrocarbons in methanogenie aquifier
material: A microcosm study. Environ Sci Technol 20(10):997-1002.

Wilson RH. 1942. Benzene poisoning in industry. J Lab Clin Med 27:1517-
1521 (as reported in Goldstein 1977).

Windholz M, Budavari S, Blumetti RF, Otterbein ES, eds.  1983. The Merck
Index, An Encyclopedia of Chemicals, Drugs, and Biologicals. 10th ed.,
p. 151. Rahway, NJ: Merck and Co., Inc.

Windholz M. Budavari S, Stroumtsos LY, Fertig MN. eds. 1976. Merck
Index, 9th ed. Rahway,  NJ: Merck and Co. Inc. (also reported in HSDB
1987).

Winek CL, Collom WD, Wecht CH. 1967. Fatal benzene exposure by glue
sniffing. Lancet (March 25):683.

Winek CL, Collom WD. 1971. Benzene and toluene fatalities. J Occup Med
13(1):259-261.

Withey RJ, Hall JW. 1975. The joint toxic action of perchloroethylene
with benzene or toluene in rats. Toxicology 4:5-15

-------
                                                        References   167

Withey RJ, Martin L. 1974. A sensitive micro method for the analysis of
benzene In blood. Bull Environ Contain Toxicol 12(6) :659-664.

Witz G, Rao GS, Goldstein BD. 1985. Short-term toxicity of trans,
trans-mucondialdehyde. Toxicol Appl Pharmacol 80:511-516.

* Wolf MA, Rowe VK, McCollister DD, Hollingsworth RL, Oyen F. 1956.
Toxicological studies of certain alkylated benzenes and benzene. AMA
Arch Ind Health 14:387-398.

* Wong 0, Morgan RW, Whorton MD. 1983. Comments on the NIOSH Study of
Leukemia in Benzene Workers. Technical report submitted to Gulf Canada,
Ltd, by Environmental Health Associates, August 31 (as reported in EPA
1986).

Yin SN, Li GL, Tain FD, Fu ZI, Jin C, Chen YJ, et al. 1987a. Leukemia in
benzene workers: a retrospective cohort study. Br J Ind Med 44:124-128.

Yin SN, Li Q, Liu Y, Tian F, Du C, Jin C. 1987b. Occupational exposure
to benzene in China. Br J Ind Med 44:192-195.

Young RJ, Rinsky RA, Infante PF, Wagoner JK.  1978. Benzene  in consumer
products. Science  199:248

Zenz C. 1978. Benzene--Attempts to establish  a lower  exposure standard
in the United States. A review. Scand J Work  Environ Health 4:103-113
(as reported in  Cole and  Merletti  1980).

Zhang  Z,  Dong G.  1982. Sister  chromatid exchanges  in plants.  I  Ch'uan
Hsueh  Pao  (J Genet)  9:357-362.

Zhurkov VS, Fel'dt EG, Kosyakov W.  1983.  Dependence of the frequency  of
chromosomal aberrations  in mouse  bone marrow cells on concentration
 (dose) and mode  of administration of benzene. Bull Exp Biol Med (USSR)
96:1741-1743.

Zini  C,  Alessandri M.  1967.  Anomalia leucocitaria pseudo-pelgeriana in
un caso  di emopatia benzolica con leucosi acuta terminale. Haematologica
 52:258-266 (as reported in Goldstein 1977).

 Zoloth SR, Michaels DM,  Villalbi JR. Lacher M. 1986. Mortality among
 commercial pressmen. J Natl Cancer Inst 76:1047-1051.

 Zwolinskl BJ,  Wilhoit RC. 1971. Handbook of Vapor Pressures and Heats of
 Vaporization of Hydrocarbons and Related Compounds. API-44-TRC101.
 College Station, TX: Ther©dynamics Research Center (as reported in
 CHEMFATE 1987).

-------
                                                                     169
                             11.  GLOSSARY

Acute Exposure--Exposure to a chemical for a duration of 14 days or
less, as specified in the Toxicological Profiles.

Bioconcentration Factor (BCP)--The quotient of the concentration of a
chemical in aquatic organisms at a specific time or during a discrete
time period of exposure divided by the concentration in the surrounding
water at the same time or during the same time period.

Carcinogen--A chemical capable of inducing cancer.

Ceiling value (CL)--A concentration of a substance that should not be
exceeded, even instantaneously.

Chronic Exposure--Exposure to a chemical for 365 days or more, as
specified in the Toxicological Profiles.

Developmental Toxicity--The occurrence of adverse effects on the
developing organism that may result from exposure to a chemical prior to
conception (either parent), during prenatal development, or postnatally
to the time of sexual maturation. Adverse developmental effects may be
detected at any point in the life span of the organism.

Embryotoxicity and Fetotozicity--Any toxic effect on the conceptus as a
result of prenatal exposure to a chemical; the distinguishing feature
between the two terms is the stage of development during which the
insult occurred. The terms, as used here, include malformations and
variations, altered growth, and in utero death.

Frank Effect Level (PEL)--That level of exposure which produces a
statistically or biologically significant increase in frequency or
severity of unmistakable adverse effects, such as irreversible
functional impairment or mortality, in an exposed population when
compared with its appropriate control.

EPA Health Advisory--An estimate of acceptable drinking water levels  for
a chemical substance based on health effects  information. A health
advisory is not a legally enforceable  federal standard, but serves as
technical guidance to assist federal,  state,  and  local officials.

Immediately Dangerous to Life or Health  (IDLH)--The maximum
environmental concentration of  a contaminant  from which one could  escape
within 30 min without any escape-impairing symptoms or  irreversible
health effects.

-------
170   Section 11

Intermediate Exposure--Exposure to a chemical for a duration of 15-364
days, as specified in the lexicological Profiles.

Imnunologic Toxicity--The occurrence of adverse effects on the immune
system that may result.from exposure to environmental agents such as
chemicals.

In vitro--Isolated from the living organism and artificially maintained,
as in a test tube.

In vivo--Occurring within the living organism.

Key Study—An animal or human toxicological study that best illustrates
the nature of the adverse effects produced and the doses associated with
those effects.

Lethal Concentration(LO) (LCLO)--The lowest concentration of a chemical
in air which has been reported to have caused death in humans or
animals.

Lethal Concentration(SO) (LCso)--A calculated concentration of a
chemical in air to which exposure for a specific length of time is
expected to cause death in 50% of a defined experimental animal
population.

Lethal Dose(LO) (LDLO)--The lowest dose of a chemical introduced by a
route other than inhalation that is expected to have caused death in
humans or animals.

Lethal Dose(50) (LDSO)--The dose of a chemical which has been calculated
to cause death in 50% of a defined experimental animal population.

Lowest-Observed-Adverse-Effect Level (LOAEL)--The lowest dose of
chemical in a study or group of studies which produces statistically or
biologically significant increases in frequency or severity of adverse
effects between the exposed population and its appropriate control.

Lowest-Observed-Effect Level (LOEL)--The lowest dose of chemical in a
study or group of studies which produces statistically or biologically
significant increases in frequency or severity of effects between the
exposed population and its appropriate control.

Malformations--Permanent structural changes that may adversely affect
survival, development, or function.

Minimal Risk Level—An estimate of daily human exposure to a chemical
that is likely to be without an appreciable risk of deleterious effects
(noncancerous) over a specified duration of exposure.

Mutagen--A substance that causes mutations. A mutation Is a change in
the genetic material in a body cell. Mutations can lead to birth
defects, miscarriages, or cancer.

-------
                                                          Glossary   171

Neurotoxlcity--The occurrence of adverse effects on the nervous system
following exposure to a chemical.

No-Observed-Adverse-Effect Level (NOAEL)--That dose of chemical at which
there are no statistically or biologically significant increases in
frequency or severity of adverse effects seen between the exposed
population and its appropriate control. Effects may be produced at this
dose, but they are not considered to be adverse.

No-Observed-Effect Level (NOEL)--That dose of chemical at which there
are no statistically or biologically significant increases in frequency
or severity of effects seen between the exposed population and its
appropriate control.

Permissible Exposure Limit (PEL)--An allowable exposure level in
workplace air averaged over an 8-h shift.

q *--The upper-bound estimate of the low-dose slope of the dose-response
curve as determined by the multistage procedure. The q.* can be used to
calculate an estimate of carcinogenic potency, the incremental excess
cancer risk per unit of exposure (usually pg/L for water, mg/kg/day for
food, and /Jg/nr* for air).

Reference Dose (RfD)--An estimate (with uncertainty spanning perhaps an
order of magnitude) of the daily exposure of the human population to a
potential hazard that is likely to be without risk of deleterious
effects during a lifetime. The RfD is operationally derived from the
NOAEL (from animal and human studies) by a consistent application of
uncertainty factors that reflect various types of data used to estimate
RfDs and an additional modifying factor, which is based on a
professional judgment of the entire database on the chemical. The RfDs
are not applicable to nonthreshold effects such as cancer.

Reportable Quantity (RQ)--The quantity of a hazardous substance that is
considered reportable under CERCLA. Reportable quantities are:  (1) 1 Ib
or greater or (2) for selected substances, an amount established by
regulation either under CERCLA or under Sect. 311 of the Clean Water
Act. Quantities are measured over a 24-h period.

Reproductive Toxicity--The occurrence of adverse effects on the
reproductive system that may result from exposure to a chemical. The
toxicity may be directed to the  reproductive organs and/or the  related
endocrine system. The manifestation of such toxicity may be noted as
alterations in sexual behavior,  fertility, pregnancy outcomes,  or
modifications in other functions that are dependent on the integrity of
this system.

Short-Term Exposure Limit  (STEL)--The maximum concentration to  which
workers can be exposed for up  to IS min  continually. No more  than four
excursions are allowed per day,  and there must be at least 60 min
between exposure periods. The  daily TLV-TWA may not be exceeded.

-------
172   Section 11

Target Organ Toxlclty--This term covers a broad range of adverse effects
on target organs or physiological systems (e.g., renal, cardiovascular)
extending from those arising through a single limited exposure to those
assumed over a lifetime of exposure to a chemical.

Teratogen--A chemical that causes structural defects that affect the
development of an organism.

Threshold Limit Value (TLV)--A concentration of a substance to which
most workers can be exposed without adverse effect. The TLV may be
expressed as a TWA, as a STEL, or as a CL.

Time-weighted Average (TWA)--An allowable exposure concentration
averaged over a normal 8-h workday or 40-h workweek.

Uncertainty Factor (UP)--A factor used in operationally deriving the RfD
from experimental data. UFs are intended to account for (1) the
variation in sensitivity among the members of the human population,
(2) the uncertainty in extrapolating animal data to the case of humans,
(3) the uncertainty in extrapolating from data obtained in a study that
is of less than lifetime exposure, and (4) the uncertainty in using
LOAEL data rather than NOAEL data. Usually each of these factors is set
equal to 10.

-------
                                                                     173
                         APPENDIX:  PEER REVIEW

     A peer review panel was assembled for benzene.  The panel consisted
of the following members: Dr. E. P. Cronkite, Brookhaven National
Laboratory (retired); Dr. Carroll A. Snyder, New York University Medical
Center; and Dr. Robert Snyder. State University of New Jersey, Rutgers
Campus. These experts collectively have knowledge of benzene's physical
and chemical properties, toxicokinetics, key health end points,
mechanisms of action, human and animal exposure, and quantification of
risk to humans. All reviewers were selected in conformity with the
conditions for peer review specified in the Superfund Amendments and
Reauthorization Act of 1986, Section 110.

     A joint panel of scientists from ATSDR and EPA has reviewed the
peer reviewers' comments and determined which comments will be included
in the profile. A listing of the peer reviewers' comments not
incorporated in the profile, with a brief explanation of the rationale
for their exclusion, exists as part of the administrative record for
this compound. A list of databases reviewed and a list of unpublished
documents cited are also included in the administrative record.

     The citation of the peer review panel should not be understood to
imply their approval of the profile's final content. The responsibility
for the content of this profile lies with the Agency for Toxic
Substances and Disease Registry.

-------