United States        Office of Health and      EPA-600/8-82-003
Environmental Protection    Environmental Assessment   January ' 982
Agency          Washington OC 20460
Research and Development
Health Assessment
Document for
1,1,1 -Trichloroethane
(Methyl Chloroform)

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                                                 EPA-60U/8-8J-003

                                                     Review Draft
                        DRAFT
                Do not cite or quote
            HEALTH ASSESSMENT DOCUMENT
                       FOR
               1,1,1-TRICHLOROETHANE
                (METHYL CHLOROFORM)
                       Notice

This document is a preliminary draft.   It has not
been formally released by EPA and should not at
this stage be construed to represent Agency
policy.   It is being circulated for comment on
its technical accuracy and policy implications.
       U.S.  ENVIRONMENTAL PROTECTION AGENCY
        Office of Research and Development
   Environmental Criteria and Assessment Office
   Research Triangle Park, North Carolina  27711

      Project Coordinator:  Dr. Jean C. Parker

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                                  DISCLAIMER







     The report Is an Internal draft for review purposes only and does not



constitute Agency policy.   Mention of trade names or commercial products does



not constitute endorsement or recommendation for use.
                                       11

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                                    PREFACE
     The Office of Health and Environmental Assessment, 1n consultation with
an Agency work group, has prepared this health assessment to serve as a "source
document" for Agency-wide use.  Originally the health assessment was developed
for use by the Office of Air Quality Planning and Standards, however, at the
request of the Agency Work Group on Solvents, the assessment scope was expanded
to address multimedia aspects.  This assessment will help insure consistency in
the Agency's consideration of the relevant scientific health data associated with
methyl chloroform.
     In the development of the assessment document, the scientific literature
has been inventoried, key studies have been evaluated and summary/conclusions
have been prepared so that the chemical's toxicity and related characteristics
are qualitatively identified.  Observed effect levels and other measures of
dose-response relationships are discussed, where appropriate, so that the
nature of the adverse health responses are placed in perspective with observed
environmental levels.

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                              TABLE OF CONTENTS
LIST OF TABLES	       vi i
LIST OF FIGURES	        ix
                                      «
1.     SUMMARY AND CONCLUSIONS	       1-1

2.     INTRODUCTION	       2-1

3.     GENERAL BACKGROUND INFORMATION	       3-1
      3.1 CHEMICAL AND PHYSICAL PROPERTIES, ANALYTICAL METHODOLOGY	       3-1
          3.1.1  Chemical and Physical Properties	       3-1
          3.1.2  Analytical Methodology	       3-3
                 3.1.2.1 Sampling and Sources of Error	       3-5
                 3.1.2.2 Calibration	       3-8
                 3.1.2.3 Standard Methods	      3-10
      3.2 PRODUCTION,  USE, AND EMISSIONS	      3-10
          3.2.1  Production	      3-10
          3.2.2  Usage	      3-12
          3.2.3  Emissions	      3-14
      3. 3 ATMOSPHERIC  TRANSPORT, TRANSFORMATION AND FATE	      3-14
          3.3.1  Residence Time and Tropospheric Removal Mechanisms...      3-14
          3.3.2  Impact Upon the Ozone Layer	      3-21
          3.3.3  Laboratory Studies	      3-26
      3.4 AMBIENT MIXING RATIOS	      3-27
          3.4.1  Global  Atmospheric Distributions	      3-27
      3.5 REFERENCES	      3-36

4.     METABOLIC FATE AND DISPOSITION 	       4-1
      4.1 ABSORPTION,  DISTRIBUTION AND ELIMINATION	       4-1
          4.1.1  Oral  and Dermal Absorption 	       4-1
          4.1.2  Pulmonary Uptake and Body Burden	       4-3
          4.1.3  Tissue Distribution	       4-13
          4.1.4  Pulmonary Elimination	       4-15
          4.1.5  Elimination by Other Routes	       4-17
      4. 2 BIOTRANSFORMATION 	       4-17
          4.2.1  Magnitude of Methyl Chloroform Metabolism	       4-18
          4.2.2  Kinetics of Blood and Urine Metabolites	       4-21
          4.2.3  Enzyme Pathways of Methyl Chloroform
                 Metabolism	       4-26
      4. 3 SUMMARY AND CONCLUSIONS	       4-32
      4.4 REFERENCES	       4-34

5.     TOXIC EFFECTS	       5-1
      5.1 HEALTH EFFECTS IN HUMANS	       5-1
          5.1.1  Experimental Studies	       5-1
          5.1.2  Occupational Studies	       5-8
          5.1.3  Accidental Exposure	       5-11

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                               CONTENTS (cont.)
      5.2 EFFECTS ON ANIMALS	      5-15
          5.2.1  Acute and Subacute Effects.;	      5-15
          5.2.2  Central  Nervous System Effects	      5-21
          5.2.3  Cardiovascular Effects	      5-21
      5.3 TERATOGENICITY, MUTAGENICITY, AND CARCINOGENICITY	      5-29
          5.3.1  Teratogenicity, Embryotoxicity,  and Reproductive
                 Effects	      5-30
                 5.3.1.1   Overview	      5-30
                 5.3.1.2   Human Studies	      5-33
                 5.3.1.3   Animal Studies	      5-33
                          5.3.1.3.1  Rats	      5-33
                          5.3.1.3.2  Mice	      5-35
                          5.3.1.3.3  Chicken embryos	      5-35
          5.3.2  Mutagenicity	      5-36
          5.3.3  Carcinogenicity	      5-38
      5.4 SUMMARY OF ADVERSE HEALTH EFFECTS AND LOWEST OBSERVED
          EFFECTS LEVELS	      5-43
          5.4.1  Inhalation Exposure	      5-43
                 5.4.1.1  Effects of Single Exposures	      5-44
                 5.4.1.2  Effects of Intermittent or Prolonged
                         Exposures	      5-50
          5.4.2  Oral Exposure	      5-53
          5.4.3  Dermal Exposure	      5-55
      5. 5 REFERENCES	      5-56

6.     BIBLIOGRAPHY	       6-1

7.     APPENDIX:  THE CARCINOGEN ASSESSMENT GROUP'S CARCINOGEN
      ASSESSMENT OF METHYL CHLOROFORM	       7-1

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                                LIST OF TABLES
3-1  Physical properties of 1,1,1-trichloroethane	      3-1
3-2  Major producers of methyl  chl oroform	      3-11
3-3  Stabilizers used in methyl chloroform formulations	      3-13
3-4  1978 emission losses to air 	      3-15
3-5  Relative efficiency of halocarbons in reducing
     stratospheric ozone	      3-24
3-6  Important atmospheric reactions that affect stratospheric
     ozone	      3-25
3-7  Ambient air mixing ratios  of CH3CC13 measured at sites
     around the world	      3-33

4-1  Partition coefficients of  methyl chloroform and other
     sol vents at 37°C	      4-2
4-2  Estimated uptake of MC during a single 4-hr exposure
     (average body weight, 77 kg; 67 kg lean body mass),
     estimated uptake of MC during a single 8-hr exposure
     (average body weight, 74 kg)	      4-8
4-3  Mean values and SEM for 12 male subjects at rest and
     exercise for 30-minute periods	      4-10
4-4  Tissue content (rat) of methyl chloroform (MC) after
     chronic inhalation exposure at 500 ppm	      4-14
4-5  Recovery Experiment with rats (3) intraperitoneally
     injected with 14C-MC(700 mg/kg)	      4-20
4-6  Relation between inhalation exposure and urinary metabolites
     of MC	      4-22

5-1  Subjective and physiological responses to a constantly
     increasing methyl chloroform vapor concentration over
     a period of 15 mi nutes	      5-4
5-2  Subjective and physiological reponses to methyl chloroform
     vapor concentrations of 900 to 1000 ppm	      5-5
5-3  Number of subjective responses to methyl chloroform
     exposure	      5-7
5-4  Signs and symptoms of patients surviving intoxication with
     methyl chloroform	      5-12
5-5  Acute toxicity of methyl chloroform	      5-16
5-6  The relative hepatotoxic efficacy of chlorinated solvents	      5-21
5-7  Probable result of single exposure to the vapors of methyl
     chloroform	      5-22
5-8  Left ventricular and hemodynamic effects of
     methyl chloroform 	      5-25
5-9  Concentration of chemicals causing cardiac sensitization
     and their physical properties	      5-27
5-10 Oncogenic and terotogenic testing of methyl chloroform	      5-31
5-11 Summary of NCI chloroethane bioassay results as of
     July 1978	      5-39
                                      vn

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                            LIST OF TABLES (cont.)

                                                                      Page

5-12 Summary of neoplasms in rats and mice ingesting
     methyl chloroform 	     5-41
5-13 Summary of neoplasms in rats inhaling methyl  chloroform
     for 52 weeks	     5-42
5-14 Human fatalities associated with methyl chloroform
     inhalation	     5-45
5-15 Non-lethal effects of methyl chloroform on humans 	     5-47

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                            LIST OF FIGURES


                                                                      Page

3-1  Global distribution of methyl chloroform	     3-30

4-1  Absorption and pulmonary elimination of MC,  and blood
     concentration	     4-6
4-2  Data from one subject exposed to 30-min periods of MC	     4-11
4-3  Postulated pathways of hepatic biotransformation of MC	     4-27
                                      ix

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The authors of this document are:
Dr. Richard Carchman, Department of Pharmacology, The Medical College of
     Virginia, Health Sciences Division, Virginia Commonwealth University,
     Virginia

Dr. I.  W. F. Davidson, Department of Physiology/Pharmacology, The Bowman Gray
     School of Medicine, Wake Forest University, Winston Salem, North Carolina

Mr. Mark M. Greenberg, Environmental Criteria and Assessment Office, U.S.
     Environmental Protection Agency, Research Triangle Park, North Carolina

Dr. Jean C. Parker, Environmental Criteria and Assessment Office, U.S. Environ-
     mental Protection Agency, Research Triangle Park, North Carolina
Assistants to the Project Coordinator:

Mr. Mark M. Greenberg, Environmental Criteria and Assessment Office, U.S.
     Environmental Protection Agency, Research Triangle Park, North Carolina.

Ms. Donna Sivulka, Environmental Criteria and Assessment Office, U. S.
     Environmental Protection Agency, Research Triangle Park, North Carolina.

Dr. David E. Weil, Environmental Criteria and Assessment Office, U. S. Environ-
     mental Protection Agency, Research Triangle Park, North Carolina.

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The following Individuals were asked to review an early draft of this
document and submit comments:

Or. Joseph Borzelleca
Dept.  of Pharmacology
The Medical College of Virginia
Virginia Commonwealth University
Richmond, VA  23298

Dr. Benjamin Van Duuren
Institute of Environmental Medicine
New York Univeristy Medical Center
New York, NY  10016

Dr. Herbert Cornish
Dept.  of Environmental and Industrial Health
University of Michigan
Ypsilanti, MI  48197

Dr. I. W. F. Davidson
Dept.  of Physiology/Pharmacology
The Bowman Gray School of Medicine
Winston-Sal em, NC  27103

Dr. Lawrence Fishbein
National Center for Toxicological Research
Jefferson, AR  72079

Dr. John G. Keller
P. 0.  Box 10763
Research Triangle Park, NC  27709

Dr. John L. Laseter
Director, Environmental Affairs, Inc.
New Orleans, LA  70122

All Members of the
Interagency Regulatory Liason Group
Subcommittee on Organic Solvents
                                       x1

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The following individuals attended a review workshop to discuss draft
EPA documents on organic compounds which included an early draft of
this document:

Dr. Mildred Christian
Argus Laboratories
Perkasie. PA  18944

Or. Rudolf Jaeger
Institute of Environmental Medicine
New York, NY  10016

Dr. Benjamin Van Duuren
Institute of Environmental Medicine
New York University Medical Center
New York, NY  10016

Dr. Herbert Cornish
School of Public Health
University of Michigan
Ann Arbor, MI  48197

Dr. I. W. F. Davidson
Dept. of Physiology/Pharmacology
The Bowman Gray School of Medicine
Winston-Sal em, NC  27103

Dr. John Egle
Dept. of Pharmacology
Virginia Commonwealth University
Richmond, VA  23298

Dr. John Keller
P. 0. Box 10763
Research Triangle Park, NC  27709

Dr. Norman Tn'eff
Dept. of Preventive Medicine
University of Texas Medical Branch
Galveston, TX  77550

Dr. Thomas Haley
National Center for Toxicology Research
Jefferson, AK  72079

Dr. James Withey
Food Directorate
Bureau of Food Chemistry
Ottawa, Canada
                                       xii

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Participating members of the Carcinogen Assessment Group

Roy E. Albert, M.D. (Chairman)
Elizabeth L. Anderson, Ph. D.
Larry D. Anderson, Ph. D.
Steven Bayard, Ph. D.
Chao W. Chen, Ph. D.
Bernard H. Haberman, D. V. M. , M.S.
Charaligayya B. Hiremath, Ph. 0.
Chang S. Lao, Ph. D.
Robert McGaughy, Ph. D.
Beverly Paigen, Ph. D.
Dharm V. Sinch, D.V.M, Ph. D.
Nancy A. Tanchel, B.A.
Todd W. Thorslund, Sc. D.
Participating members of the Reproductive Effects Assessment Group

Peter E. Voytek, Ph. D. (Chairman)
John R. Fowle III, Ph. D.
Members of the Agency Work Group on Solvents

Elizabeth L. Anderson         Office of Health and Environmental Assessment
Charles H. Ris                Office of Health and Environmental Assessment
Jean Parker                   Office of Health and Environmental Assessment
Mark Greenberg                Office of Health and Environmental Assessment
Cynthia Sonich                Office of Health and Environmental Assessment
Steve Lutkenhoff .             Office of Health and Environmental Assessment
James A. Stewart              Office of Toxic Substances
Paul Price                    Office of Toxic Substances
William Lappenbush            Office of Drinking Water
Hugh Spitzer                  Consumer Product Safety Commission
David R. Patrick              Office of Air Quality Planning and Standards
Lois Jacob                    Office of General Enforcement
Arnold Edelman                Office of Toxic Integration
Josephine Breeder             Office of Water Regulations and Standards
Mike Ruggiero                 Office of Water Regulations and Standards
Van Jablonski                 Office of Solid Waste
Charles Delos                 Office of Water Regulations and Standards
Richard Johnson               Office of Pesticide Programs
Priscilla Holtzclaw           Office of Emergency and Remedial Response
                                     xin

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                          1.  SUMMARY AND CONCLUSIONS







     1,1,1-Trichloroethane  (methyl  chloroform,  MC)  is  a  chlorinated  hydro-



carbon compound which is manufactured and used in large quantities.   Since its



commercial introduction  in  the mid-1900s, it has been used increasingly as an



industrial solvent and in consumer products, such as spot removers.   In recent



years, the U.S.  production  of MC has grown significantly; current production



is estimated  to  have increased from 266 million pounds (121  million  kg)  in



1970 to 716 million pounds (325 million kg) in 1979.  The compound is  produced



by three  manufacturers with a total  annual  production capacity of 975 million



pounds.  Methyl  chloroform's  popularity  is  apparently due primarily to early



studies which indicated that it had a very low toxicity in comparison to other



halogenated hydrocarbon solvent's" with similar physical properties.



     It is estimated that approximately  88 percent of the MC consumed in the



United States  is lost directly  to  the environment through dispersive use,



largely by evaporation to the atmosphere.  Three-quarters of the air emissions



are attributed to  metal  cleaning operations which  consume 66 percent of  the



total  production.  There  are  no known natural sources of MC.   In addition to



the workplace, ambient air and water measurements indicate that it is found in



a variety of  urban and non-urban areas of the United States and other regions



of the world.  Measurable amounts of MC  have been  reported in the atmosphere,



soil,  rainwater, marine  and fresh surface waters,  and ground water  in the low



part per  billion (ppb)  level.  Residues of  MC have been measured at similar



levels (low ppb) in the tissues of aquatic and terrestrial plants and animals,







002MC1/H                             1-1                               11-10-81

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and in food items.   In addition,  MC has been detected in human expired air,  at
levels of 0.03-140 ug/h/ subject.  Based on available information, an average
                                                   3
ambient air concentration of  about I ppb (5.4 ug/m  ) would  be expected for
some large urban centers, while  air samples from rural areas average around
100 parts per trillion (ppt).
     Chlorine atoms are released  from MC by photodissociation in the strato-
sphere and react with  ozone (0-) thus contributing to a reduction of strato-
spheric Og.   Therefore, MC may contribute to the effects of ozone depletion on
human health, i.e., an increased incidence of certain  types  of  skin cancers
due to increased UV-B  radiation  reaching the earth's surface.   However, the
approximate extent to which stratospheric ozone is depleted by all halocarbons
and the extent to which past,  current and future emissions of MC contribute to
this depletion can  only  be  estimated through computer  modeling  of the  stra-
tosphere.  It is  not  currently possible to verify by direct measurement an
ozone depletion of less than 2 percent per decade.  Unlike chlorofluorocarbons,
which are not  known to be  removed by mechanisms  in  the troposphere, MC is
removed  to a substantial extent  through  reaction  with hydroxyl  radicals.  The
tropospheric lifetime  of MC is generally assumed  to  be  within a range of 5  to
12 years  in  spite  of the lack of scientific agreement  on  the global  average
concentration of hydroxyl radicals.   It  has been estimated that  this period of
time allows  from  10 to 20  percent  of the MC emitted to survive tropospheric
removal  mechanisms  and reach  the stratosphere.   The National  Academy of
Sciences  has recently  estimated  that MC currently contributes one-quarter to
one-half  the number of chlorine  atoms reaching  the stratosphere, as  do  either
of  the chl orofl uorocarbons  F-ll  or  F-12.   Assuming  the same  rates  of  pro-
duction,  MC  has been  calculated to be  approximately 15  and 20 percent as
 002MC1/H                              1-2                               11-10-81

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effective  as chlorofluorocarbon F-ll and  F-12,  respectively,  in destroying
ozone.  This estimate assumes a 9-year tropospheric  lifetime  for MC.   If  it
replaces other  solvents  in the future, or as  new uses for MC are found, it
will necessarily play a more significant role in ozone photochemistry.
     Methyl  chloroform production  has been rising in recent years, and esti-
mates  of  tropospheric levels  show a corresponding rate  of  increase of 17
percent annually.   The most  recent  measurements indicate a global  average
mixing ratio of about 110 ppt.
     At levels of methyl chloroform necessary for anesthesia (10,000 to 50,000
ppm),  significant  cardiovascular effects  have been observed  in  animals and
humans.  Even at  1000 ppm, MC inhalation  produces cardiovascular effects  in
humans that  include sensitization of the heart to spontaneous or catecholamine-
provoked arrhythmias  and'hypotension.  This cardiovascular  response is, in
part,  the  result of  MC's  ability  to depress the conduction  system  of the
heart.
     More  recent studies have  also shown that MC is not entirely innocuous to
human  health at the  lower concentrations  likely to  be encountered  in the
                  *
workplace  (TLV-TWA  = 350  ppm),  although  single exposures to  concentrations
less than  5000  ppm are probably not  potentially life threatening to humans.
The  lowest observed adverse  effect  lever (LOAEL,  500 - 350 ppm) produces
subjective symptoms of  lightheadedness,  syncope, stuffiness, mild headache,
*
 TLV-TWA (Threshold  Limit  Value  -  Time  Weighted Average):  is defined as the
time-weighted average concentration  for a  normal 8-hour workday and a 40-hour
workweek to which nearly all workers may be exposed repeatedly, day after day,
without adverse effect.
 LOAEL:  is defined as the lowest exposure level in a study or group of studies
which produces statistically significant increases in frequency or severity of
adverse effects between  the exposed population and its appropriate control.
002MC1/H                             1-3                              11-10-81

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nausea, and mild irritation of eye,  nose,  and throat.   No significant abnormal
blood chemistry or organ  function  tests have been noted.  The  most adverse
effects are  neurological  symptoms, which have  been  diagnosed by subjects'
impaired performance  of cognitive and manual  tasks.
     The incidence of adverse health effects at low ambient air mixing ratios
(~ 100 ppt)  is  unknown.   However,  because there is evidence that MC accumu-
lates in the  body, long-term or lifetime exposure to  even  low ambient air
concentrations might represent  a  health hazard.  A no observed effect level
     *
(NOEL ) for  long  term  occupational  exposure of humans to MC  is  presently
assumed to be around  53 ppm.
     Methyl chloroform has been demonstrated to have mutagenic activity in the
Ames test, with and  without metabolic activation, and in cultured  mammalian
cell transformation  systems.   If  the metabolism and pharmacokinetics of this
compound in  humans result in metabolic products which can interact with DNA,
as is the case for bacteria, it may cause mutagenic effects in humans as well.
It may also possibly have teratogenic potential, thereby showing similarity to
other halogenated hydrocarbon t:ompounds.   In tests conducted to date, MC has
not  appeared  to be carcinogenic;  however, the  fact  that it is mutagenic in
Salmonella warrents further examination of its carcinogenic potential.
     The pharmacokinetics and metabolism of MC  have been studied in man.  Like
other halogenated hydrocarbon  solvents, inhalation and lung absorption of MC
vapor  in  the air  is  the most important and  rapid  route of  absorption into the
body.   It  is estimated that for an 8  hour exposure at the TLV-TWA  (350 ppm),
about 2 grams will be absorbed  into the body of a normal 70 kg man.  The  total
 NOEL:   is  defined as the exposure level at which there are no statistically
significant  increases  in frequency or severity of effects  between the exposed
population and its appropriate control.
 002MC1/H                             1-4                               11-10-81

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amount absorbed increases  in  direct  proportion to inspired air concentration
and to length  of  exposure and physical activity.  However,  at equilibrium,
only approximately 30 percent of the inspired air concentration is retained.
Because of its lipophilic nature, MC  is expected to cross membrane barriers in
the body  and  diffuse  into the brain  and the milk of nursing mothers,  as well
as  into  the  fetus during pregnancy.   There  is strong evidence that tissue
concentration of MC,  particularly into adipose tissue, will occur with chronic
or  long  term  exposure to  even low ambient air  concentrations.  Methyl chloro-
form is  metabolized  in  man to a very  limited  extent  (about  6  percent of the
total retained dose).
     The  primary  route  of elimination  of MC  is via the  lungs.  The only  iden-
tified uninary  metabolites are  trichloroethanol  and trichloroacetic acid.
Metabolism of MC  is  enhanced by other  chemicals  and  drugs  such as phenobar-
bital, and there  is  some evidence that MC  may enhance its  own metabolism.
     As stated above, some of the health hazards of exposure to MC concern the
                               »«•
possible  mutagenic, teratogenic,  and carcinogenic potential of the compound.
The extent to  which  its possible carcinogenic  potential  can be realized is
linked to the  biochemical mechanisms of its metabolism in the liver.   Methyl
chloroform may  be biotransformed to  "reactive  intermediate"  metabolites  which
may  contribute  to tissue  and organ  toxicities, as well  as  carcinogenesis.
However,  the  evidence  for reactive  intermediate  metabolites  of  MC is frag-
mentary  at present.   Methyl  chloroform has been demonstrated  to be mutagenic
in  the Ames  test, with and without  metabolic  activation, and in a cultured
mammalian cell  transformation system.   However, animal  bioassays  performed  by
the National  Cancer  Institute (NCI)  and others have  not provided definitive
evidence  of carcinogenicity.   At the present time, there  is  inadequate evidence
to  properly  evaluate  MC as a  chemical  carcinogen (see  attached  CAG report).

002MC1/H                             1-5                              11-10-81

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     There are only a few studies on the teratogenicity and fetotoxicity  of  MC
and the  limitations  of  the available data do not allow for a  full  assessment
of the effects.   Further testing is required to determine whether this  toxicity
is a  health hazard to  humans,  particularly  at low ambient concentrations.
     Until there is  considerably more information available on the biological
effects of MC and  its role in contributing to an increased incidence of  skin
cancer as  a  result of ozone depletion, a  definitive  evaluation  of all  the
health hazards  associated  with  its use in the workplace  and  its presence in
the ambient  environment is not  possible.   For example,  there is very  little
data on  synergistic  or  antagonistic  effects  of MC with other  compounds;  e.g.,
anesthetic  or  vasoactive  agents.   In  addition,  a lifetime animal bioassay
under  the  National  Toxicology Program using both  rats and mice  is currently
nearing  completion.   It should  be noted that the MC being used in this  study
has only a very low percent (0.002%) of dioxane, a stabilizer thought possibly
to contribute to  the positive  results  obtained  in the mutagenicity  and  cell
transformation  tests.          *
 002MC1/H                             1-6                              11-10-81

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                           2.   INTRODUCTION







     1,1,1-Trichloroethane (methyl  chloroform,  MC) belongs  to  a family of



saturated chlorinated compounds widely  used in industrial cold cleaning and



vapor degreasing operations,  in  the synthesis of vinylidene chloride,  and  in



consumer products.   This document provides an evaluation of the  health  hazards



of MC and a review of the relevant scientific literature.   In order to  provide



a perspective  in evaluating  the  health hazards of MC, this document contains



background chapters  relating  to analytical  methodologies, production, sources



and emissions, ambient air concentrations, and levels of exposure.



     Methyl chloroform  is released  into the ambient air as a result of evapo-



ration during  production,  storage,  and manufacturing or  during general con-



sumer use.  It is  not known  to* be  derived from natural sources.   In recent



years, a  great deal  of attention has been  focused upon the role of MC  in the



destruction of  stratospheric  ozone, indirectly resulting in increased  ultra-



violet  radiation  at the  earth's  surface  and contributing  to  an increased



incidence of  skin  cancer.  MC is  one  of the many  atmospheric pollutants whose



reactivity is  sufficiently  low to allow  it to  be transported to the strato-



sphere;  its trophospheric  lifetime  is estimated  to be  5  to 12 years.   Con-



centrations in  ambient  air  sampled around  the nation reflect methods used to



control  emissions  and  the transport and  transformation processes  in the at-



mosphere.



     The scientific  literature on MC is limited in reference to its effects on



humans.   There are  relatively few  epidemiological studies.   What  is known









002MC1/D                            2-1                          11-10-81

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about the  effects  of MC has been primarily  learned  from studies involving

individuals exposed  occupationally or accidentally to it.  In such exposures,

the concentrations of MC associated with adverse effects to human health were

either unknown or far in excess of the concentrations measured in ambient air.
                                                             *
Controlled exposure  studies  have  been carried out at TLV-TWA  concentrations

(350 ppm)  and  have been directed toward elucidation  of  the  pharmacokinetic

parameters of  MC exposure.   These studies have established that  vapor inhala-

tion is  the  principal  route by which MC enters  into the body; it is widely

distributed  into all organ  systems and  is metabolized to a limited but signi-

ficant extent.  MC is  eliminated from  the body primarily as  the parent  com-

pound via  the  lungs, but metabolites  are excreted mostly in  the  urine.   Given

evidence  that  MC  accumulates  in the body,  particularly  in  fatty tissues,

long-term  or lifetime exposure to even low  ambient air concentrations  may

represent  significant  health hazards.   Epidemiological  studies  provide  some

information  about  the  impact of MC on  human health, but it  is necessary to
                                «.
rely on  animal studies to  derive  indications of  potential  harmful  effects for

chronic  low  exposure.  These studies are reviewed in chapters 4  and  5.

     Apart from the  important questions of carcinogenicity and teratogenicity,

other human  biological effects relate  to the narcosis potential of  MC and to

evidence  that  low ambient concentrations may  have  accumulative  neurological

and cardiovascular effects.  The  full detrimental consequences to  human  health

of  MC  have not been determined  at  present and require  further study.  These

questions  are  discussed in  chapter 5.
  TLV-TWA (Threshold Unit Value - Time Weighted  Average):   is defined as the
 time-weighted average  concentration  for a normal 8-hour  workday  and a 40-
 hour workweek to  which nearly all workers may be exposed  repeatedly,  day
 after day,  without adverse  effect.
                                     2-2

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     The potential of  MC  or its metabolites to  act  as  chemical  carcinogens
and/or mutagens represents  a most serious hazard to human health.  This docu-
ment summarizes the  available mutagenicity studies and  the  animal  bioassay
studies that relate  to the  carcinogenic potential of MC.  Methyl chloroform's
potential  to cause human  cancer is discussed in chapter 5.   According to the
EPA Carcinogen  Assessment Group  and  Reproductive  Effects Assessment Group
Reports (January, 1981),  it remains suspect at  the present  time; however, a
final  judgment on the  carcinogenicity of MC has been deferred until  further
information becomes available, particularly the results of the current National
Toxicology Program bioassay in rats and mice.
     Permissible levels of MC in the working environment have been established
in various countries.  The  U.S. Occupational Safety and  Health Administration
(OSHA) health standard  requires  that a worker's exposure  to  MC at no time
exceed an 8 hr time-weighted average o
air in any work shift of a 40 hr week.
exceed an 8 hr time-weighted average of 1,900 mg/m  (350 ppm) in the workplace
                                    2-3

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                        3.   GENERAL BACKGROUND INFORMATION



3.1  CHEMICAL AND PHYSICAL PROPERTIES, ANALYTICAL METHODOLOGY

3.1.1  Chemical and Physical Properties

     1,1,1-Trichloroethane (CHgCCl,), also called methyl chloroform (MC), is a

colorless nonflammable  liquid which  has  a characteristic odor.   Its line

formula is:

                                   H  Cl
                                   i   i
                                 H-C—C-C1
                                   i   i
                                   H  Cl


Chemical Formula CpHjCl.,

Chemical Abstracts Service Registry Number 71-55-6

     Synonyms and Identifiers

     Aerothene TI                       Inhibiso]
     Chloroethane NU                    Methyl Chloroform
     Chiorotene                         Methyltri chloromethane
     Chlorothane NU                     NCI-C04626
     Chlorothene                        Alpha-T.
     Chlorothene NU                     Trichloroethane
     Chlorothene VG                     1,1,1-Trichloroethane
     Chlorten                           orTrichloroethane
     Ethane, 1,1,1-Trichloro


     Table 3-1 shows some of its important chemical and physical properties.

             TABLE 3-1.   PHYSICAL PROPERTIES OF 1,1,1,-TRICHLOROETHANE


Solubility in water @ 25°C	0.44 gm/lOOgm
Boiling point @ 760 torr	74°C
Vapor pressure @ 20°C	100 torr
Vapor density (air = 1)	4.6
Molecular weight	133.41  -
1 ppm	5.4 mg/m



002MC1/G                           3-1                           3-25-82

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     In the atmosphere, MC is subject to free radical  attack and reaction with
hydroxy] radicals  is  the principal  way  in  which it is scavenged  from the
atmosphere.  Photooxidation products  of  MC  include hydrogen chloride, carbon
oxides, phosgene, and acetyl chloride (Christiansen et al., 1972).  The prin-
cipal tropospheric photooxidation product has  been reported to be trichloro-
acetaldehyde (U.S. Environmental Protection Agency, 1975).  Data discussed in
Section 3.3 indicate that MC is relatively stable in the troposphere  and that
significant amounts are conveyed to the stratosphere.
     In water, methyl chloroform is  slowly hydrolyzed to predominantly acetic
and hydrochloric acids (Billing et  al., 1975).   Billing et al.  (1975) reported
a half-life of hydrolysis of 6 months at 25°C (dark reaction).
     Anhydrous MC is  generally  noncorrosive,  but in the presence of water it
can react  to  form hydrochloric  acid, which is  a  corrosive of metals (Keil,
1979).   The  addition  of epoxides can  neutralize  the  generated acid  (Keil,
1979).   Anhydrous MC,  when heated to 360° to 440°C, decomposes to 1,1-dichloro-
ethylene and hydrogen chloride.   When MC is  heated in the presence of water at
temperatures between  75° and 160°C,  it decomposes, upon contact with  metallic
chlorides  or  sulfuric  acid,  to  acetyl chloride, acetic acid,  and  acetic an-
hydride.   Noweir  et al.  (1972)  have observed that when  MC comes in contact
with iron, copper,  zinc,  or aluminum at elevated  temperatures,  phosgene is
produced.
     Bonse and Henschler (1976) have considered the general chemical  reactivity
of MC.  By virtue of  the electron-inductive effects of the chlorine  substitu-
ents,  destabilization  occurs at the C-C bond.   Radical  reaction mechanisms
would be favored  as  a result of the delocalization of the unpaired  electron
drawn from the C-C bond.
002MC1/G                           3-2                           3-25-82

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3.1.2  Analytical Methodology
     lo detect very  low levels of methyl chloroform in ambient air, sophisti-
cated analytical  techniques have been employed.   The two most generally useful
methods for detection  and  analysis  of MC have  been  gas  chromatography with
electron capture detection  (GC-ECD)  and gas chromatographymass spectrometry
(GC-MS).  Both systems have a lower limit of detection on the order of a few
parts per trillion (ppt).   Ihe utility of GC-ECD over GC-MS  is that it can be
used in the field  to provide nearly continuous measurements through  inter-
mittent sampling (every 15 to 20 minutes).
     In a comparison between GC-ECD and GC-MS, Cronn  et al. (1976) judged
GC-ECD to be superior  in precision.  Standards  for four  halocarbon compounds,
measured by GC-ECD,  had  coefficients of variation  ranging  from 0.5 to  3.5
percent.  Of 11  halocarbon  compounds measured by GC-MS,  coefficients of  vari-
ation ranged  from  4 to 19  percent,   temperature  programming or isothermal
operation of the gas chromatograph  yielded comparable results (Cronn et al.,
1977).   The precision with  temperature-programming was 4.3 percent versus 3.5
percent with isothermal operation for MC.
     A close agreement between the levels of MC and other halocarbons detected
by both GC-ECD and  GC-MS  (on the same  ambient  air samples) was obtained by
Russell  and Shadoff  (1977).   A difference of 10 ppt was reported between the
systems.  Air samples  were  sorbed onto Porapak N porous polymer and desorbed
onto the column  by heating  the collection tube  rapidly to 200°C.  Ambient air
mixing ratios measured by  this method were  in  the  range of 90 to 110 ppt.
     Ihe electron capture  detector,  as well as the mass spectrometer in the
selective ion monitoring mode  (Cronn and Harsch, 1979a;  Pellizzari, 1974),  is
specific in the  detection  and quantisation of many halogenated hydrocarbons,
whereas nonhalogenated hydrocarbons  are not detected.   Thus, a high background

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level of  hydrocarbons  in  ambient air samples does  not  preclude analysis of
trace quantities of MC.   In the electron capture detector, MC  is ionized by
primary electrons  released from an  internal radioactive  source.   The  net
result is removal  of  electrons  from the gaseous mixture with substitution by
negative  ions of greater  mass.   The measured effect is  a net decrease in ion
current because the mobility  of the ions is less than that of the electrons.
     Samples of tropospheric  air were analyzed by Cronn and coworkers (1972)
both by preconcentrating  the  samples according to the method of Rasmussen et
al.  (1977)  and  by direct  injection.   The detection limit was  3  ppt.   The
precision of the method  using preconcentration was ±4 percent.   The internal
consistency between direct GC-ECD  analysis  of  MC  and using the preconcen-
tration method was  reported to be good (Cronn et al., 1977).
     A detection limit of 2 ppt for MC was achieved by Harsch and Cronn (1978)
with a low  pressure sample-transfer  technique.  Precision was ±7 percent with
the  Rasmussen  et  al.   (1977)  preconcentration  technique.   Calibration  was
accomplished by use of secondary standards that were compared to static dilu-
tions of  pure and  commercially  prepared mixtures of  MC (10  ppm) in helium.
Robinson  (1978) reported  a detection limit of 6 ppt with a precision of ±4.2
percent with secondary standards in a system using dual  GC-ECD.
     Pellizzari  and Bunch  (1979) reported  an estimated detection limit of
                   _c      2
12.45 ppt (66 x 10   mg/m )  using  high resolution gas chromatography/mass
spectrometry.   The detection  limit  was calculated on the basis of the break-
through volume for 2.2  grams  of Tenax GC, at 21°C.   Ihe accuracy of analysis
was  reported  at ± 30  percent.   Sources of  error are discussed in  section
3.1.2.1.   Water
     The relatively high volatility of MC was used to advantage by Piet et al.
(1978) in measuring the  content of  MC  in water samples.   A direct headspace


002MC1/G                           3-4                           3-25-82

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method was used with  MC  volatilizing into the air above  the  surface  of the
water sample in a  sealed container.   A GC-ECD system was  used for separation
and quantitation.   Ihe systematic error of the method was reported to be less
than 5 percent.   Ihe detection limit for MC was 0.05  ug per  liter.   Ihe  0V  225
capillary GC column provided good separation of MC from CCK.
3.1.2.1   Sampling  and Sources of  Error—Ihe  National  Academy of  Sciences
(1978) has reviewed common approaches used to sample  ambient air for trace  gas
analysis.  These approaches include:
1.   Ambient pressure samples.  An evacuated chamber  is opened and allowed to
     fill until it has reached ambient pressure at the sampling location.  If.
     filling is conducted  at high altitude, contamination of the low pressure
     sample is likely when samples are returned to ground level.
2.   Pump pressure samples.  A  mechanical  pump is used to  fill  a stainless
     steel or glass container to a positive pressure  relative to the surround-
     ing atmosphere.
3.   Adsorption on molecular sieves,  activated charcoal,  or other materials.
     Sorbents that have  been  used to collect MC include Porapak Q, lenax GC,
     and Chromosorbs 101, 102,  and 103.
4.   Cryogenic sampling.    Air samples  are transferred to a loop held at the
     temperature of liquid oxygen.  Components, including MC, are concentrated
     by freezeout while other gases (oxygen, nitrogen) pass  through.
     In the  analysis  of  whole  air samples of  compounds such  as MC which are
present  in ambient  air  in the ppt range,  small  uncontrolled errors in the
system could result  in  inaccurate results.  Errors in the analysis can occur
during sampling, in  the  gas chromatograph, in the EC detector, and in  cali-
bration of the instruments.
002MC1/G                           3-5                           3-25-82

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     Rasmussen and  Khali!  (1981)  reported that, in an  inter!aboratory  com-
parison of two samples  of  MC in ambient  air,  a majority of the 19 partici-
pating laboratories reported values lower than those determined by the authors.
It was concluded that common standards are needed if atmospheric measurements
from different laboratories are to be pooled.
     Singh and coworkers (1978a)  used  electrochemically-polished stainless
steel  sampling vessels.  The sampling vessels were flushed with ultra-pure
helium until  background  contamination was reduced to less than  2  to 3 percent
of the expected background  concentration  of a  given trace constituent.  Cronn
and  coworkers similarly  employed  electropolished stainless steel  containers
whose  interior surfaces  had been passivated by  electropolishing (Cronn and
Robinson, 1978;  Cronn et al., 1976; 1977).
     Harsch and Cronn (1978) have  evaluated sample  transfer techniques.  Most
analytical procedures  involve  the  collection  of pressurized  air samples.
Sample transfer under positive pressure minimizes serious contamination possi-
bilities  posed  by  laboratory air  containing  many  times the ambient  mixing
ratios of most halocarbons.  However, collection of low pressure samples, such
as  stratospheric  air samples, is  a specialized situation  in which careful
precautions against sample contamination are  necessary.  A technique appli-
cable  to  collection of  whole air  samples at  altitudes up  to 21 kilometers
showed a  7  percent standard deviation when  the MC  mixing  ratio was  105 ppt
(Harsch  and  Cronn, 1978).   Electropolished  sampling bottles were evacuated to
30  microns  prior  to sample  collection.   Transfer of  contents  to a freezeout
loop (Rasmussen  et  al., 1977)  (immersed in liquid 02)  was made  by  vacuum
assist.   Preconcentration  was  followed by injection  onto  a temperature-pro-
grammed  column.   The detection limit for MC  with  the GC-ECD procedure was 2
ppt.   To minimize the number of potential sources  of leaks, system components


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were silver  soldered.   Ihe technique was  applicable  when gross halocarbon
levels were  not  present  in laboratory air.  However,  it  was  noted that if
laboratory air contained  ppb  levels  of halocarbons,  high-boiling halocarbons
were adsorbed onto the surfaces of the fittings of the sampling containers  and
associated plumbing during  times  when the system was  exposed  to laboratory
air.
     The EC  detector  can  be a source of error because water vapor and oxygen
may cause  reduced sensitivity for certain halocarbons.   Lillian and Singh
(1974) used  an ascarite trap between  the GC column and the detector to absorb
sample moisture.   Carrier gas  for the GC is purified by passage  through traps
containing activated charcoal, anhydrous  calcium sulfate, and an ascarite  or
molecular sieve  (Cronn,  1980).   Oxygen contamination in  the detector can be
minimized by preconcentration  of  air samples on porous polymers (Russell  and
Shadoff, 1977).   Significant  oxygen  contamination is often caused by fitting
leaks in the GC rather than by carrier gas contamination.
     A freezeout  concentration method was  employed by  Sasmussen  et al. (1977)
to  determine atmospheric  levels  of MC in the presence of other trace vapors.
The detection  limit of  MC was reported to be  0.8 ppt  for 500  ml aliquots of
ambient air  when  measured by GC-ECD.  Precision was 4.3  percent.  Standards
were prepared  by  static  dilutions in helium.   During the procedure,  the oven
of  the GC was  cooled  to -  10°C.  When  freezeout was complete,  the loop con-
taining the  concentrated air  sample was   immersed in  heated  water and the
carrier gas swept the  contents of the sample loop onto the column.
     Pellizzari and Bunch (1979) reported the use of Tenax GC, a porous polymer
based on  2,6-diphenyl-p-phenylene oxide,  to  absorb MC  from  ambient air.
Recovery was made by  thermal  desorption and  helium purging  into a freezeout
trap.   Included  among the inherent analytical errors were (1) the ability to


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accurately determine the breakthrough  volume,  (2)  the percent recovery from
the sampling cartridge after a period of storage, and (3) the reproducibility
of thermal desorption from the cartridge and its introduction into the analy-
tical  system.  To minimize  loss  of sample, cartridge samplers should be en-
closed  in  cartridge holders and placed  in a second container that  can  be
sealed,  protected from light,  and stored at 0°C.
3.1.2.2    Calibration—In  general,  calibration of  the instrumentation  for
GC-ECD analysis of  methyl  chloroform  has involved  static,  multiple dilutions
of pure material  in the  ppm range to  ppt  levels (Cronn  et al.,  1976;  1977;
Harsch and Cronn, 1978; Lillian and Singh, 1974; Singh et al., 1977a).   Singh
et al.  (1977b) have  cautioned that,  in some cases, multiple  dilutions  of
materials to ppt  levels  are  tedious  and prone to inaccuracies.  In order to
overcome the difficulties in generating low-ppb primary standards of MC,  Singh
et al.,  (1981) has  reported that permeation tubes  offer the most accurate
means.  Permeation  tubes were  conditioned  for  at least two weeks and,  during
standard generation, were  kept in holders  maintained at 70 ± 0.1°C.  Error in
the permeation rate (980  mg/min) were ± 15 percent.  A reproducible means of
generating low-ppb  primary standards was  judged essential  since there are
considerable difficulties  in  storing  long-term secondary standards.   Various
investigators have used GC-ECD systems to quantitate MC in ambient air samples
(Cronn et  al. ,  1976;  1977; Lillian and  Singh, 1974; Rasmussen et  al., 1977;
Singh et  al. ,  1977c).  An  inter!aboratory  calibration  (Cronn et  al., 1976) of
six real  ambient  air  samples gave a precision  (percent standard  deviation) of
11.4  ppt.   A similar comparison calibration was reported  by Singh et al.,
(1981).
      Rasmussen and  Khali!  (1981) reported  that primary standards of  MC (~ 139
and 52 ppt) remained stable for up to about one year, even though the pressure
in the master  tanks had decreased significantly.

002MC1/G                           3-8                           3-25-82

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        Because the ionization  efficiency  of  MC in the EC  detector  is only 20

   percent, use of  dual  EC detectors in series have been  used to achieve higher

   accuracy (Lillian and Singh,  1974).   The dual detection system, as employed by

   Singh et al. (1977c),  enables  MC to be quantitated coulometrically according

   to the following equation:

             Coulombs  = 96,500 pW;
   where
                    p  = ionization efficiency
   and              W  = moles  of compound

   Determination of the ionization efficiency can be made  by use of the following

   expression which relates p  to  the signals  (X,  and  X2) of two  identical EC

   detectors in series:

                      X
              P =  I-X^

             X1 =  96,500 pW

             X2 =  p (96.500W - 96,500 pW)

   The  ionization efficiency  should be determined for the  operating  conditions

   and once established, can be used in the coulometric calibration.

        The accuracy of the GC-ECD approach has been reported by Singh (1977a) to

   be  better  than 10  percent for  compounds  such as MC that  have ambient air
1C
   mixing  ratios  of 20 ppt or greater.  An overall system accuracy of ± 10 per-

   cent was reported  by Singh et al.  (1977a)  in analyses of MC in ambient air

   samples.  Two  gas  chrotnatographs,  each equipped with  two  EC  detectors,  were

   employed.  An  ascarite  trap  was placed between  the GC column and  the  EC de-

   tector  to prevent  moisture interference.  Precision was  reported to be better

   than 5  percent and instruments were calibrated  using  multiple dilutions of

   pure material.    Singh et  al.,  (1981) recently  reported linearity of dual

   frequency-modulated  ECD's  over a wide concentration range for MC, down to

   low-ppt levels.


   002MC1/G                           3-9                           3-25-82

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3.1.2.3  Standard Methods—The analytical method S335, suggested by NIOSH for
organic solvents in  air,  utilizes  adsorption on charcoal  followed by desorp-
tion with carbon disulfide.   The resulting effluent is analyzed by gas chroma-
tography.   This method was  recommended for the range 96 to 405 ppm.  Inter-
ferences are minimal  and  those  that do occur  can  be  eliminated by altering
chromatographic conditions.   However,  one  disadvantage  is that the charcoal
may suffer breakthrough, thus limiting the amount of air that can be  sampled.
3.2  PRODUCTION, USE, AND EMISSIONS
     Methyl chloroform is principally  used for the cold  cleaning  and vapor
degreasing of  fabricated metal  parts.   Because  of  its  volatility and dis-
persive use pattern,  much  of the MC produced  worldwide  is emitted into the
atmosphere.  There are no  identified natural  sources of emissions.   Once in
the atmosphere, MC is subject to atmospheric transport and transformation.   To
better gauge the effects  of present and future emissions of MC on human health,
this section profiles MC production, usage, and emissions.
3.2.1  Production
     Methyl chloroform is produced by several manufacturing processes (Jordan,
1979;  Lowenheim and Moran, 1975; U.S. Environmental Protection Agency, 1979b):
     1.   Hydrochlori nation   of  1,1-dichloroethylene  (vinyl idene  chloride)
                                EeCi,
               CH2 =  CC12 +  HC1 - i*   CH3CC13
     2.  Hydrochlori nation of vinyl chloride
                                Fed.
               CH£ =  CHC1 +  HC1 - i*   CH3CHC12
                         .
               CH3CHC12  g    > CH3CC13 + HC1
      3.   Chlorination of ethane
                CH3CH3 + 3C12  370°  to 480°C>   CH3CC13 +  3HC1 + byproducts
 002MC1/G                            3-10                           3-25-82

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     In order to prevent  catalyst deactivation,  hydrochlorination reactions

should be anhydrous  (Jorden,  1979).   Temperature control of thermal  reactors

for chlorination must be maintained to ensure MC  stability (Jordan, 1979).   In

reaction (3)  above,  ethyl  chloride and vinylidene chloride  are byproducts.

Increased yields of MC can be obtained through recycling of these  byproducts.

     In 1975, over 60  percent of the U.S. production of MC was derived from

the hydrochlorination of  vinyl  chloride;  derivation  from vinylidene chloride

accounted for 30 percent (Lowenheim and Moran,  1975).

     In 1977, the U.S. production figure  for MC was 288,565 metric tons (U.S.

Environmental Protection Agency,  1979b;  U.S.  Internationa] Trade Commission,

1977).  The major producers and their production  capacities for 1977 are shown

in Table 3-2.   Quoting  production estimates supplied by Dow Chemical U.S.A.,

Singh and coworkers  (1979a)  reported that production of the chlorocarbon has

been  increasing and,  in  recent years, this  increase  has been at the annual

rate of 12  percent.   Estimates did not take  into  account production in the
     TABLE 3-2.   MAJOR PRODUCERS OE METHYL CHLOROFORM1". (Cogswell, 1978)
                                                1978 capacity
     Organization                               metric tons

     Dow Chemical, Freeport, IX                  204,000

     PPG Industries, Inc., Lake Charles, LA       79,000

     Vulcan Materials Co., Geismar, LA            29,000
 Production capacity estimated to increase significantly at PPG Industries
and at Vulcan Materials facilities for 1982 (Chemical and Engineering News,
1979; Chemical Marketing Reporter, 1979; Chemical Week, 1978).
002MC1/G                           3-11                          3-25-82

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Soviet Union and other eastern European countries.  The Dow Chemical estimate
for global production in 1977 was 470,000 metric tons.  The annual production
growth rate through  1977 was  reported to be 7 to 10 percent by trade sources
(Chemical and Engineering News,  1979).   SRI  Internationa]  estimated an average
production growth between  5.5 and 7.5 percent annually up  to 1982.   Future
growth in production depends on  the status of other chlorinated solvents under
regulation for which MC may be substituted.
3.2.2  Usage
     In 1975, cold  cleaning and  vapor degreasing operations accounted for 75
percent of the total end use of MC.  Approximately 12 percent was  used  in the
synthesis of vinylidene chloride  (Lowenheim and Moran, 1975).  Use  in  metal
cleaning operations  was  reported  to  total  67  percent of 1977 production
(Cogswell,  1978).
     MC is also used as a solvent in adhesive formulations,  as a spot remover,
as a film cleaner,  and  as  an additive  in metal  cutting oils (Jordan, 1979;
Keil, 1979;  Lowenheim and  Moran,  1975).   Stabilizers used in MC are shown in
Table 3-3 and protect it in the  following ways (Jordan, 1979):  (1) prevent or
retard oxidation, (2) chelate metal ions, (3) scavenge HC1, and (4)  passivate
metal surfaces.  Vapor degreasing grades  of MC contain 3 to  7 percent  (w/w)
stabilizers  and additives (Jordan, 1979);  principal stabilizers include nitro-
methane, 1,3-dioxolane, 1,4-dioxane,  butylene  oxide,  sec-butyl alcohol, iso-
butyl alcohol, N-methy1 pyrrole, and toluene (Jordan,  1979).   In film cleaning
applications, a low  concentration of alcohol is used  as a stabilizer (Jordan,
1979).
     SRI International reported that between 11,000 and 13,000 metric tons  of
MC are  used  annually in  personal  care products  (Cogswell,  1978).    Expected
growth in this area is 15 to 20  percent annually (Cogswell,  1978).


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         IABLE 3-3.   STABILIZEfiS USED IN MEIHYL CHLOROFORM FORMULATIONS
                                 (Jordan, 1979)
     1,4 dioxane
     1,3-dioxolane
     butylene oxide
     epichlorohydrin
     methyl ethyl ketone
     sec-butyl alcohol
     isobutyl alcohol
     ethyl acetate
     N-methylpyrrole
     ter-amylphenol
     hydroquinone monomethyl-
       ether
     methylal
     ethylene glycol mono and
       dialkyl ethers
               aery 3 on i *.
               p-ethoxypropioni tri1e
               diallylamine
               di i sopropy1 ami ne
               acetaldoxime
               nitromethane
               nitroethane
               N-methylmorpholine
               Thymol
002MC1/G
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3.2.3  Emissions
     Emissions of MC  arise  during primary and end product production, during
dispersive use applications,  from storage containers, and during disposal of
waste materials.
     Singh et al. (1979c) have estimated global emissions in 1977 at 95 percent
of the  300,000 metric  tons  produced.   Lovelock (1977a) estimated that 500,000
metric tons were released globally in 1975.  Ihe estimate of the global emissions
rate in  1976  by  the National  Research  Council  (1979)  was  about 439,000 metric
tons.  According to McCarthy  (1975), almost  all  the MC produced  is  eventually
released to the atmosphere.
     Recent estimates  in a  report prepared for the U.S.  Environmental  Protection
Agency, Office of loxic Substances, placed 1978 emission  losses to air  at 214,000
metric tons (Katz et al. 1980).   Emission estimates by sources category are  shown
in Table 3-4.   Ihe  WMO (World Meteorological  Organization, 1982), using  data
reported by Neely and  Agin  (U.S.  E.P.A., 1980), estimated current global  annual
releases at 476,000 metric  tons.
3.3.  ATMOSPHERIC TRANSPORT,  TRANSFORMATION  AND EAT.E
     Ihe potential for ambient air mixing  ratios of MC to pose a hazard to human
health  is  influenced  by the  many processes  that occur in the troposphere and
stratosphere.  Such processes include:  transformation of MC into other poten-
tially  harmful  atmospheric  components; urban transport;  tropospheric chemical
reactivity; and  diffusion into the stratosphere where MC  participates in  ozone
(0,) destruction reactions.
3.3.1   Residence Time  and Iropospheric Removal Mechanisms
     Scientists  generally agree  that the  longer the  tropospheric residence  time
for  a  chemical  species, the greater  the likelihood of diffusion  into the  strato-
sphere.   Concern that MC is destroying stratospheric 03  and thus increasing  the
 incidence  of  certain  forms  of skin  cancer (Chapter 5) has intensified recently.

 002MC1/G                           3-14                          3-25-82

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                    TABLE 3-4.  1978 EMISSION LOSSES TO AIR
            Source
   1978 emission losses to air in
      million pounds (kkg) and
 percentage of total released to air
Production
     1. From Vinyl Chloride            0.21
     2. From Vinylidene Chloride       0.15
     3. From Ethane                    0.03
Metal Cleaning                       351.70
Aerosols                              39.90
Adhesives                             38.37
Textiles                               6.44
Paints                                10.91
Inks                                   6.13
Drain Cleaners                         0.61
Pharmaceuticals                        0.27
Film Cleaning                          0.48
Leather Tanning                        0.23
Catalyst Preparation                   0.06
Miscellaneous                         14.82

     Total released to the air*      470
                     (95)
                     (67)
                     (14)
                (159,500)
                 (18,100)
                 (17,400)
                  (2,920)
                  (4,950)
                  (2,780)
                    (278)
                    (124)
                    (218)
                    (104)
                     (28)
                 (16,730)
 0.04%
 0.03%
 0.01%
75%
 8%
 8%
 1%
 2%
 1%
 0.1%
 0.06%
 0.1%
 0.05%
 0.01%
 3%
                (213,300) (100%)
*An additional 63 million "pounds (28,602 kkg) are released to the
 environment via solid waste and water.
 Adapted from Katz et al. (1980).
002MC1/G
3-15
  3-25-82

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     A recent conference convened by the U.S. Environmental Protection Agency



(1980) met to establish  the facts about the distribution and persistence of



MC.   The  results of research  in atmospheric chemistry related to the question



of stratospheric 0, depletion by  MC were discussed.  Presentations from the



various perspectives  of  those  involved  in,  or  concerned  with,  regulatory



decision-making were also made.



     Mass budget and other  modeling estimates suggest a  longer residence time



than was previously predicted.  Before 1977,  it was commonly believed that the



residence time was  in  the range of 1 to 2 years  (National  Research Council,



1976). Such  a  time  span  suggested a tropospheric  reactivity great enough to



preclude  any serious  impact on 03 levels.  The  "short"  lifetime for MC was



predicated a global average tropospheric hydroxyl radical   (OH) concentration


                     55             -3
in the  range 10  x 10  to 30  x 10  molecules cm   .  Reaction with OH is the



principal tropospheric removal mechanism by which many compounds are scavenged



from the  atmosphere.   The following reactions, occurring in the troposphere,



would serve  as a  sink  for MC,  thus  reducing the  levels prior to  its diffusion



to the stratosphere:




                  CH3CC13 + -OH 	•	>  -CH2CC13 + H20      (1)




                  CH3CC13 + 0(1D)	  -CH2CC13  +  -OH  (2)




                  [0(1D) =  atomic oxygen in its  singlet  state ]




      Indications  of the  extent to which MC  is  calculated  to  deplete  strato-



spheric 0,  are  predominantly derived from one-  and two-dimensional  modeling
          •3

studies.  The following  scenarios consider projections of  atmospheric loading,



residence time and photochemical processes in the  atmosphere.
 002MC1/G                            3-16                           3-25-82

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     Campbell (U.S.  Environmental Protection Agency, 1980)  has calculated the



rate of  consumption  of MC via oxidation by OH as a  function of altitude and



latitude.  One-half  of the oxidation occurs below 2.4  km.  Most of the oxi-



dation also  takes  place in the tropics, i.e., half  the  removal of MC occurs



between  16°N and 16°S.  These modeling  efforts  agreed with measured hemi-



spheric ratios and suggested a mean lifetime for MC of 6.4 years.



     Logan et al.,  (1981)  modeled MC concentrations over time and latitude,



using altitude profiles of OH averaged over an annual cycle.  The model  results



lend support to the findings of Campbell (U.S.  Environmental Protection Agency,



1980) in that MC removal is particularly sensitive to OH levels in the tropics,



a region that accounts for about 70 percent of the global sink.   Approximately



50 percent of the  global sink occurs over the tropical ocean.  By integrating



MC loss  over altitude  and  latitude,  using current reaction  rate constants for



key reactions,  Logan  et al.,  (1981) calculated a global lifetime for MC of 5



years.    It was  emphasized  that  one-dimensional  models are  inappropriate for



calculating the lifetime of a compound such as MC.   The model  of Logan et al.,



(1981) predicted a mean OH level  in the northern hemisphere of 1 x 10  per cm .


                                                       5      3
This value is  somewhat higher than the  level (2.5 x 10  per cm ) calculated



by Crutzen and  Fishman (1977).   In the model  studies of Derwent and Eggleton



(1981),  a global  lifetime  for MC was calculated at  4.8  years, using a two -



dimensional   model.   These  investigations  found  that global or hemispheric



models were inapproriate; in the case of MC, a lifetime of 12 years was  calcu-



lated when OH was averaged over all  altitudes,  latitudes, and seasons.   It was



concluded that  neglect of the variations  of  OH  destruction with altitude,



latitude, and season may lead to an under - or overestimation of the impact of MC



on stratospheric 03.  Global and box models, as evaluated by Derwent and  Eggleton,



gave larger  0-  depletion  estimates; 1-D models  underestimate such impact.









002MC1/G                           3-17                          3-25-82

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     The mass balance studies of Singh (1977a;  1977b) had predicted reduced OH
levels (3 x  10  molecules cm  ) to account for observed ambient levels of MC.
                                                                      c
These OH levels are in reasonable agreement with the OH values (3 x 10  to 4 x
10 ) determined by  Campbell  and coworkers (1979) during monitoring studies.
From the hemispheric  distribution  of MC, OH abundance in the northern hemi-
sphere was  1.5 to  3  times  lower than in the  southern  hemisphere.   Carbon
monoxide (CO)  levels  were believed primarily responsible for the  lower north-
ern hemisphere levels of OH.   However, Campbell (U.S. Environmental Protection
Agency, 1980)  has  cautioned  that,  because of the combination of MC oxidation
in the  tropics  and CO reduction in  the  northern hemisphere, there  is  a dif-
ficulty with using the N/S ratio of MC to obtain hemispheric means of OH.   For
example, a  southern  hemisphere OH  concentration  several times higher than
northern hemisphere values  requires the transition  from the southern hemi-
sphere value to  the lower northern  hemisphere  value to  occur within  a few
degrees of  the equator.   Based  upon  the  observed tropospheric distribution  of
MC,  atmospheric  loading,  and its rate of  reaction  (National  Aeronautics  and
Space  Administration,  1977)  with OH, Singh  (1977a;  1977b)  computed a global
average residence time between 8 and 11 years.   This time span would be suffi-
cient  to allow 15 to 22  percent  of the  MC. emitted  to  reach the stratosphere
(National Aeronautics and Space Administration, 1977; Singh et al.,  1979a).
The  National Research Council  (1979a),  using 1976  global release rates,  esti-
mated  that  about 12 percent  reaches  the  stratosphere.
      Singh  and coworkers  (1979b) have found that field measurements support a
6  to 8 year global  average residence time.   Measurements of MC  between latitudes
35°N and  35°S indicated a global  average concentration of 95 ppt.  When esti-
mates  of  other investigators are taken into account, the suggested range is 6
to 12 years (Singh et al.,  1979b).   This range would allow an estimated 12 to
25 percent  of all  MC  emissions  to  enter  the  stratosphere (Singh et al., 1979b).

002MC1/G                            3-18                           3-25-82

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     Ihese latter estimates agree with McConnell and Schiff (1978), who used a
one-dimensional model to compute a global average MC mixing ratio of 90 ppt, a
value consistent with observed ambient levels at the time.   The average tropo-
spheric OH  level  needed to account for  this  computed  MC level was 5  x 10
molecules cm  .
     Based upon  a MC residence time of 5.4 years, Derwent and  Eggleton (1978)
calculated that  approximately  10 percent survives tropospheric removal pro-
cesses to reach  the  stratosphere.   Lovelock (1977b) estimated a 5 to  10 year
residence time based on ambient measurements from 1971 to 1976.
     Support for an  8  to 12 year  residence  time for MC can be found  in the
modeling results of  Chang  and Penner (1978).  They computed an average resi-
dence time of 11.3 years (10 to 12.8 years) from a two-box model interrelating
hemispheric  abundance,  release  rates  and interhemispheric  transport  time
through continuity equations.
     Neely  and  Plonka   (1978),  in  contrast to  Singh and coworkers (1977a;
1977b) and  Crutzen and  Eishman (1977), who compute an  average  tropospheric  OH
                        5           5-3
concentration of 2 x 10 to 4 x  10 cm   ,  find their computed time-averaged  OH
                         5             -3
concentration of. 4.8 x 10  molecules cm   compatible with a residence  time for
MC of 2.6 to 4.0 years.   In their mass budget model, this OH concentration was
required to  reconcile emissions  (based on Dow Chemical production data) with
observed ambient MC  levels.   In  contrast to Singh, who used a global  ambient
MC mixing ratio of 90 ppt,  Neely and Plonka (1978) used 76 ppt.  The emissions
data were significantly higher than those data calculated by Singh.
     In a recent  review,  Altshuller (1980) suggested  an average  OH concen-
tration of  3  x 10  molecules cm   as the most reliable value at present.   He
cautioned, however, that uncertainty still exists about the extent to  which MC
reacts with OH.  Ihe residence time for  MC was  calculated to fall  in the 8  to
002MC1/G                           3-19                          3-25-82

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13 year range.    For example,  a residence time of 13 years was calculated from
the rate constant expression of Watson et al. (1977) at 265°K.  With the rate
constant expression derived by  Chang  and Kaufman (1977),  a MC residence time
of 8 years was computed.
     Basmussen and Khali! (1981b) reported that trospheric measurements of MC
made at six locations in the northern  and southern hemispheres, between January
1979 and June 1980 were consistent with an estimated lifetime  in the range of
6 to 10 years.   Average  concentrations over both hemispheres were calculated
from latitudinal profiles and  indicated that the global  atmospheric concen-
tration (from January 1979 to January  1981) increased at 6.7 ± 2.0 percent per
year.   In  a report  submitted  for publication, Khali 1  and Ramussen (1981)
determined that  the  increase  in concentration was  smaller  during  1980 than
during 1979. It was concluded that a decrease in the emissions rate has occur-
red since 1975.
     The World Meteorological Organization  (1982) recently  reviewed that data
base and found  that,  given  the uncertainties  in  release  rates and absolute
concentrations of MC, as discussed by Logan et al. (1981), the measurements of
MC appear to be consistent with a lifetime of 5 to 10 years, and with globally
                                      (-             _ O
averaged OH concentrations of ~ 7 x 10  molecules cm  .   This estimate was, in
part, based  on  redetermi nations of the rate constant for reaction  of HO with
MC  (Kurylo  et al.,  1979; Jeong and Kaufman, 1979)  and the knowledge that
release rates as given by Neely and Plonka (1978) do not account for release
in Eastern Bloc countries.
     Since  distribution  of  OH has only been estimated through use  of models,
estimates  for the atmospheric  lifetime  of MC should be  cautiously viewed.
002MCD/G                           3-20                          3-25-82

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3.3.2  Impact Upon the Ozone Layer
     As stratospheric ozone levels decrease, changes in the amount of light in
the  solar  spectrum around 300 urn that reach the earth's surface may forecast
an  increased incidence of  skin cancer  (National  Research Council,  1979b,
1982).  Reduction in 03 levels also increases the likelihood of adverse changes
in  weather and climate  (National  Research Council, 1979b, 1982).   Reduced
ozone  concentrations  could result  from the following reaction mechanisms:
                              hv
                  CH3CC13  	—	>  CH3CC12-+ Cl-      (1)
                  Cl- + 03 	*  CIO + 02            (2)
                  CIO- + 0  	>  Cl- + 0£           (3)
                  CIO- + NO 	»  Cl- + N0£          (4)
     The atomic chlorine produced in reaction (1) would react with 03 to yield
chlorine oxide.  The  subsequent chain reaction would  result  in  a continual
depletion of 0^.  The importance of this cycle and other cycles in the strato-
sphere have  recently  been evaluated by Wuebbles and Chang (1981) and the NRC
(National  Research Council, 1982).  These  evaluations  indicate that  the effi-
ciency of  the chlorine  oxide  cycle is closely coupled with  other cycles,
principally the nitrogen oxides and hydroxyl radical cycles.
     In a study (National Research Council, 1979a) of the impact of chloro and
chloroflouro compounds  on stratospheric 0-,  the National Research  Council
(NRC) concluded that  emissions  of halocarbons represented the most immediate
and serious  threat to  0,.   Based upon atmospheric measurement data available
at that time,  the  NRC concluded that MC contributes one-quarter to  one-half
the number of chlorine atoms on a molecule-per-molecule basis  that chlorofluoro-
carbons 11  and 12 contribute  to the stratosphere.  At the estimated  1976
global emissions rate  of  approximately 400,000 metric tons, MC was estimated
002MC1/G                           3-21                          3-25-82

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to destroy about 10 percent (Table 3-5) as much 03 as both chlorof1uorocarbons
11 and  12.   The magnitude of the impact of MC was not delineated further due
to a  lack  of reliable measurement data on  OH abundance in the troposphere.
The NRC  cautioned,  however, that continued  increased usage of MC could  result
in the  halocarbon  becoming the largest source of stratospheric chlorine and,
thus, the most serious threat to stratospheric 0_.
     Preliminary results  obtained using  the  Lawrence Livermore Laboratory
one-dimensional vertical  transport-kinetics model  suggest that the  relative
efficiency of  MC  in destroying ozone is  much less  than chlorof1uorocarbons
(U.S.  Environmental Protection  Agency,  1980).  Table 3-5 shows this relative
efficiency.
     The "best  estimate"  of the NRC  for the eventual steady-state 0, decrease
due to  chlorof1uorocarbons  11  and 12 at  1977 release rates was  16.5 percent
(National  Research  Council, 1979a).  This estimate was about twice as high  as
the estimate reported  by  the  NRC in 1976 (National Research Council, 1976).
Due to  the uncertainties  in the modeling approach and data base,  it was  re-
ported that there was a 95 percent probability that the true value of eventual
0^ reduction would be somewhere between 5 and 28 percent.
     In their  most  recent report,  the NRC  (1982) revised their  03 depletion
estimates  downward, principally due  to refinements of the values of  important
reaction rates.  The  NRC  now  estimates that  the  steady-state  (~ 100 years)
reduction  in total  global ozone,  in the absence of other pertubations,  could
be between 5 and 9 percent.  These estimates only considered release of chloro-
fluorocarbons 11 and 12 at the 1977 release rates.
     The World  Meteorological  Organization  (1982) assuming continued release
at the current estimated emissions rate for MC and using the model of Wuebbles
002MC1/G                           3-22                          3-25-82

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(1981) and revised reaction rate kinetics,  calculated an estimated steady-state
depletion of the total  0, column of 0.8 percent (Table 3-5).   If other strato-
spherically -important halocarbons are considered (particularly chlorofluoro-
carbon 113, carbon tetrachloride,  and  methyl  chloroform) at current release
rates, the WHO estimates  that the predicted column 03 depletion of from 5 to 9
percent would be  increased about one-third.  In scenarios more representative
of real-time conditions  for example, considering variations in carbon dioxide
and nitrogen oxides,  the  WMO derived other estimates of steady-state 0- deple-
tion.   In each scenario, a continued increase  in upper stratosphere 0, deple-
tion  is predicted.  As noted  by the  WMO                         the 5 to 9
percent depletion estimate due  to  FC-11 and FC-12,  if correct, suggests that
less  than a 1 percent decrease  in global average stratospheric 0, should have
occurred to date.  This  estimated 1 percent decrease  is not estimable be-
cause  state-of-the-art instrumentation  and statistical methodology indicate
that detection of global  average stratospheric  0-  trends is limited to about a
2 percent change per  decade.
     In addition  to the  uncertainties  and limitations of the models used to
estimate the impact of MC on 0-, the complexity of rapidly-changing knowledge
in atmospheric  chemistry make  it difficult to assess with confidence such
impact in quantitative terms.   Among the atmospheric reactions that can act to
enhance or restrict 03 depletion due to MC, the NRC (National  Research Council,
1979a) cited  those  described  in  Table 3-6 as  important determinants.   An
evaluation of the impact of MC upon stratospheric 0~ must take into account
these and other  atmospheric  processes  if realistic estimates of 0, depletion
are to be made.
002MC1/G                           3-23                          3-25-82

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             TABLE 3-5.  RELATIVE EFFICIENCY OF HALOCARBONS IN
                       REDUCING STRATOSPHERIC OZONE
               (U.S. Environmental Protection Agency, 1980)
                                                  Percent ozone depletion
               Compound                              after 320 years*
               F-ll                                         -10.7

               F-12                                          -8.5

               F-113                                         -8.25

               F-114                                         -5.38

               MC                                            -0.933

                                                             -1.6b
      *production  rate  assumed, for each species individually, at 1,000 million
       pounds  annually

      abased on 4.5 year  lifetime

       based on 9  year  lifetime
002MC1/G                           3-24                          3-25-82

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 TABLE 3-6.   IMPORTANT ATMOSPHERIC REACTIONS THAI AEFECT STRATOSPHERIC OZONE
                      (National Research Council, 1979a)
                     Cl + 03 	»   CIO + 02

                     CIO + 0 	»   Cl + 02

                     CIO + NO 	>   Cl + N0£

                     Cl + CH4 	>   HC1 + CH3

                     HC1 + OH 	»   Cl + H20

                     CIO + N02 + M 	»•   C10N02 + M

                     C10N02 + hv 	»   Cl + N03

                     CIO + H02 	»   HOC! + 02

                     HOC! + hv 	»•   Cl + OH, CIO + H
002MC1/G                           3-25                          3-25-82

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     The  British  Department  of the  Environment (1979) also  released a
report on the effect of chl orof 1 uorocarbons on stratospheric ozone.  Based on
annual production of 450,000 metric tons and an OH + MC rate constant of 19 x
10    cm  molecule   sec,    the estimated steady state ozone depletion was  1.0
percent, due to MC (Table  3-5).
3.3.3  Laboratory Studies
     In experiments designed to characterize the UV absorption spectrum for MC
in the wavelength  and  pressure ranges associated with the stratosphere, Van-
            ^
1 aethem-Meuree et al.  (1979)  found that photo-dissociation was the dominant
sink process at altitudes  above 25 kilometers.
     In Spence and Hanst's (1978) photooxidation studies, when MC was irradi-
                                     o           o
ated at maximum  intensities  of 3100 A  and 3650 A, one-fifth of the initial
amount (10 ppm) was consumed after 6 minutes.   The principal products observed
were carbon monoxide  (1.5  ppm),  hydrochloric acid (6  ppm),  and  phosgene (2
ppm):
                    Cl
         hv
   1
Cl
               H. C-C-C1  - = - >    CO + HC1  COC1
                3   i         U~
Of the halocarbons evaluated, methyl chloroform was the least reactive.
     The recent reaction rate studies, involving MC and OH, by Jeong and
Kaufman (1981) and by Kurylo et al. (1981), indicate that the rate of this
key reaction is slower and more temperature sensitive than previously had
been indicated.  Jeong and Kaufman (1981) measured reaction rates using the
discharge-flow method and MC which had been extensively purified.  A reaction
                                                -14   3    -1
rate at 293°K was determined at 1.06 ± 0.09 x 10    cm  sec  .   The Arrhenius
expression, within 95 percent confidence limits but not including systematic
errors, was 5.49 ± 1.40 x ID*12 exp [ - (1832 ± 98/T] cm3 sec"1.  The authors
002MC1/G                           3-26                          3-25-82

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indicated that the most recent rate (at 298°K) reported by JPL (Jet Propulsion
                           -14   3    -1
Laboratory, 1979), 1.9 x 10    cm  sec   is too high by a factor of about 1.7
at this temperature, and by a factor of 1.9 at 265°K, the temperature used by
Singh (1977) in deriving average OH concentrations.
     The Arrhenius expression determined by Kurylo et al., (1979) was in close
agreement with that determined by Jeong and Kaufman (1981).   Kurylo et al.
(1979) used the flash-photolysis method and also extensively purified the MC
prior to use.
3.4  AMBIENT MIXING RATIOS
     Modeling approaches for reconciling emissions of MC to ambient air mixing
                               55             -3
ratios have implied low (3 x 10  to 6 x 10  molecules cm  ) atmospheric levels
of hydroxyl radicals (OH), the principal scavenging species for MC.  If veri-
fied, these OH levels would strongly suggest that MC and many other chemical
species have a tropospheric lifetime sufficiently long to allow their signi-
ficant transport to the stratosphere.   These models, however, have relied on
point measurements of MC mixing ratios determined at disparate sites, pri-
marily in the northern hemisphere.   Before 1977, reliable estimates of glob-
ally averaged MC concentration were not available.  Recent measurements,
however, indicate that the global average concentration is consistent with a
long (5 to 11 years) tropospheric lifetime and that MC remains a serious
threat to stratospheric 03 levels.
3.4.1  Global  Atmospheric Distributions
     The global average background mixing ratio of MC has been determined by
several laboratories, including SRI International and Washington State Univer-
sity.  The field studies of Singh and co-workers at SRI have been reported in
a number of publications (Singh, 1977a; Singh et al., 1977c; 1978a; 1978b;
1978c; 1979a;  1981).  The global average for late 1977 was reported as 95 ppt
(Singh et al., 1979a).   This average was derived from measurements made in the

002MC1/G                           3-27                          3-25-82

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northern (average = 113 ppt) and southern (average = 77 ppt) hemispheres.
Based on data collected at latitudes 30°N to 40°N from 1975 to 1978,  MC pre-
sence in the atmosphere was increasing at the rate of 15 ppt (or 17 percent)
per year.
     Field studies by Singh et al.  (1979a) were conducted during two oceano-
graphic cruises between latitudes 64°N and 40°S (15 September to 30 October
1977; 20 November to 13 December 1977).   Samples were collected in electro-
polished stainless steel and glass vessels and analyzed by GC-ECD, operated
coulometrically.  Samples were analyzed ui situ as well as after storage.   Ihe
northern temperate average mixing ratio was reported as 123 ppt.  Ihe latitu-
dinal distribution is shown in Eigure 3-1.
     Ihe vertical profile and latitudinal gradients of MC in the troposphere
and lower stratosphere have been investigated by Cronn and coworkers in a
series of high-altitude studies (Cronn and Robinson, 1979; Cronn et al.; 1976;
1977a; 1977b).  Most recently, experiments conducted at the Panama Canal Zone
(9°N latitude) and at 37°N indicated that MC tropospheric abundance was 18
percent lower at 9°N (Cronn and Rqbinson, 1979).  Singh et al.  (1979a) re-
corded a 17 percent difference between these latitudes (Eigure  3-1).  For July
1977, the average tropospheric background concentration (139 whole air samples
were collected between ground level and 13.7 kilometers) was 97.3 ± 4.6 ppt at
9°N.  At 37°N (Pacific Ocean west of San  Francisco), an average mixing ratio
of 116 ± 14 ppt was measured.  Tropopause height at the Canal Zone was 15.7
kilometers.  Methyl chloroform tropospheric mixing ratios did not vary signi-
ficantly with increasing altitude at 9°N  but there was a precipitous drop  in
the mixing ratio across the tropopause.
     Analysis of pressurized tropospheric samples was  performed by a dual
GC-ECD.  Ihe detection  limit for MC was 6 ppt and the  precision of analysis


002MC1/G                            3-28                           3-25-82

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was ±4.2 percent.   Analyses were performed on 5-ml aliquots.   For the low-
pressure stratospheric air samples, precision was ±7 percent with a detection
limit of 2 ppt.  Methyl chloroform was preconcentrated with a freezeout sample
loop.  Overall accuracy of analysis was estimated at about 10 percent.
     In experiments conducted at 37°N (Cronn et al., 1977a),  samples were
collected at altitudes ranging from 6.1 to 14.3 kilometers with the tropopause
height between 11 and 12 kilometers.  Samples were analyzed by GC-ECD utiliz-
ing the freezeout concentration method.  The average trospheric background
mixing ratio for MC in April 1977 (excluding samples within 0.8 kilometer of
the tropopause) was 116 ± 14 ppt.  This value is consistent with the results
of Singh et al., (1979a), who obtained a value of 117 ppt at this latitude
during the fourth quarter of 1977 (Figure 3-1).
     When this observed mixing ratio (116 ppt) was compared with that obtained
in March 1976 at 48°N (Pacific Northwest), an annual atmospheric increase in
MC of 23.7 percent was calculated.  Cronn et al. (1976; 1977a) collected whole
air samples at altitudes ranging from 4.6 to 14.6 kilometers (average tropo-
pause height was 10.8 kilometers) during March 1976 over Western Montana and
Idaho.  The average tropospheric background mixing ratio for MC was 94.5 ±8.2
ppt.  Samples were analyzed by GC-ECD both isothermally with a 5-ml aliquot
and with the freezeout concentration method.  Precision of the overall  method
was ±4 percent.
     The Washington State University (WSU) group has made several studies by
aircraft of the latitudinal distribution of MC (Cronn, 1980a; Robinson, 1978;
Robinson and Harsch, 1978).  Interhemispheric differences of MC in the tropo-
sphere in June, 1976, were reported, with northern levels of 96 ppt and south-
ern levels of 88 ppt (Robinson and Harsch, 1978).  Lower stratospheric levels
were 80 and 67 ppt, respectively.  Work begun at WSU in 1976, and continuing


002MC1/G                           3-29                          3-25-82

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200

150

«•
ft
a
p>
d 100
o
ۥ>
I '
U
BO

.<
I
i
77ppt 113ppt

1 0
Aa _ ^. . .A .
^8v — t *
DO ,-*^; *
* Q og £ . v i
u • 0° °of. .-•"f'So

* * " * *^tf^^ft^
1
1
)0 -70 -50 -30 -10 0 10 30 BO 70 9
5 LATITUDE, degrees f
Figure 3-1.   Global  distribution of methyl  chloroform (Singh et al.,  1979a)
002MC1/G
3-30
3-25-82

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at both WSU and the Oregon Graduate Center,  has provided annual  measurements



of interhemispheric differences of MC of 1.67,  1.72,  1.56, 1.40, and 1.45 for



1975, 1976, 1977, 1978,  and 1979,  respectively (Khali!  and Rasmussen, 1980;



Robinson, 1978).   Time-trend monitoring has  been conducted nearly continuously



at a ground station in eastern Washington state since July 1977  (Cronn, 1980a).



Increases have exceeded  12 percent per year  since that time.



     Rasmussen and Khali! (1981b), from 1979 to early 1981, measured tropo-



spheric levels of MC at  six locations, ranging from 70°N to 42°S, including



several measurements at  the south pole (90°S).   Analysis was made by EC/GC.



From these measurements, a global  average concentration of 115 ppt was cal-



culated along with an annual increase of 6.7 ± 2.0 percent.  This global



average is consistent with the values reported by others for previous years.



In contrast, however, to the rate of increase reported by Singh et al. (1979a)



for the period 1975 to 1978, the rate of increase appears to be declining, which



Rasmussen and Khali 1 (1981b) attribute to an overall  reduction in the rate of



emissions.  The hemispheric difference in concentration reported by Rasmussen



and Khalil (1981b) is consistent with that reported by Singh et al., (1979b).



During the period from January 1979 to December 1980, the northern hemisphere



average was 130 ppt compared to the southern hemisphere average of 99 ppt



(Rasmussen and Khalil, 1981b).



     Intermittent monitoring of MC at various urban,  nonbackground and rural



locations has also been conducted by Cronn and coworkers (Cronn, 1980b; Cronn



and Harsch, 1979b; Cronn et al., 1979; Harsch and Cronn, 1979).   Levels greater



than 5 ppb have been measured in Claremont,  California, with an average of 1.2



ppb during September 1978 (Cronn et al., 1979; Harsch and Cronn, 1979).



Evidence of regional pollution buildup was observed in the Smoky Mountains of



Tennessee in September 1978, with levels averaging 50 percent greater than










002MC1/G                           3-31                          3-25-82

-------
background tropospheric levels typical  at that time (Cronn and Harsch,  1979;



Cronn et al., 1979).   Values in the Caucasus Mountains of the Republic  of



Georgia, USSR, in July 1979, were identical  to levels measured in rural  east-



ern Washington (Cronn, 1980b).



     A series of studies on the distribution and levels of MC and other



halogenated hydrocarbons at various sites has been conducted by Pellizzari  and



coworkers (Pellizzari, 1977; 1978; Pellizzari and Bunch, 1979; Pellizzari et



al., 1979).   Methyl chloroform was detected in the ambient air at sites in New



Jersey, New York, California, Louisiana, and Texas.  Levels are included in



Table 3-7.



     Point measurements made by Lovelock (1977b) during 1972 to 1977 resulted



in a lower global average background mixing ratio (73 ppt).  Air samples



collected at rural sites in the British Isles indicated that, during the 5



year period, MC mixing ratios increased from 31 ppt to 97 ppt.  In the south-



ern hemisphere (Africa and Antarctica), it increased from 12 ppt to 50 ppt.



Absolute accuracy of the GC-ECD method was reported as ±30 percent.



     In field measurements of ambient MC levels over North America (18°N to



65°N) from October 4 to 13, 1976, Pierotti and co-workers (1980) determined an



average tropospheric mixing ratio of 145 ± 25 ppt for this region.  The mixing



ratio dropped sharply across  the tropopause and in their stratosphere, suggest-



ing a large  sink at these altitudes.  These investigators suggested that the



data were consistent with a northern hemisphere background mixing  ratio of



about 100 ppt.  Other point measurements of MC atmospheric mixing  ratios in



the troposhere are shown in Table 3-7.
 002MC1/G                            3-32                           3-25-82

-------
                                       TABLE  3-7.  AMBIENT AIR MIXING RATIOS OF METHYL CHLOROFORM MEASURED
                                                             AT SITES AROUND THE WORLD
               Location
Type of site
Date
Maximum
 (ppb)
Minimum
 (ppb)
Average
 (ppb)
Reference
              Alaska (70°N)
CO
 i
CO
CO
Antarctica
South Pole
South Pole
South Pole
South Pole
South Pole
South Pole
South Pole
South Pole
Arizona
Phoenix
Arkansas
Helena
California
Los Angeles
Oakland
San Jose'
Point Arena
Claremont
Point Arena
Badger Pass
San Francisco

Riverside
Mill Valley


Yosemite
Palm Springs
Los Angeles
Badger Pass
Los Reyes
Stanford Hills


Los Angeles
Basin
Delaware
Delaware City

Remote
Remote
Remote
Remote
Remote
Remote
Remote
Remote

Urban

Remote

Urban
Urban
Urban
Marine
Urban
Marine
High altitude
Free tropo-
sphere
Urban
Background sub-
ject to urban
transport
High altitude
Suburban
Urban
High altitude
Marine
Background sub-
ject to urban
transport
Urban-suburban


Urban
                     Aug.  1979  -Jan.  1981                            ~ 0.155
                                                 1/75                                              0.054
                                                 1/76                                              0.057
                                                 1/77                                              0.070
                                                 10/77 - 11/77                                     0.082
                                                 1/78                                              0.083
                                                 1/79                                              0.095
                                                 Jan. 1979 - Jan. 1981   -0.12      ~  0.094
                                                 1/80                                              0.103
                                                 4/23-5/6, 1979            2.8136
                                                 11/30/76                   <  1          <  1
                     4/9-4/21,  1979            5.143
                     6/28-7/10, 1979           0.9672
                     8/21-8/27, 1978           2.931
                     8/30-9/5,  1978            0.158
                     9/78                      5
                     5/23-5/30, 1977           0.150
                     5/5-5/13,  1977            0.342

                     4/77
                     4/25-5/4.  1977            3.012
                     1/11-1/27, 1977           0.895
                                                 5/12-5/18, 1976           0.126
                                                 5/5-5/11, 1976            0.545
                                                 4/28-5/4, 1976            7.663
                                                 May 1976
                                                 12/1-12/12, 1975          0.212
                                                 11/23-11/30, 1975         0.565
                                                 4/9-4/21, 1974            2.14
                                                 7/8-7/10, 1974            0.30        0.03
                                                                   Rasmussen and Khalil, 1981b
                                                                                        Robinson, 1978
                                                                                        Ibid.
                                                                                        Ibid.
                                                                                        Ibid.
                                                                                        Ibid.
                                                                                        Khalil and Rasmussen
                                                                                        Rasmussen and Khalil, 1981b
                                                                                        Ibid.
                                                           0.1978     0.8235 ± 0.5974   Singh, et al., 1981
                                                                            < 1
                                                                   Battelle, 1977
0.224
0.1429
0.150
0.115
0.3
0.083
0.130

0.282
0.107
0.073
0.075
0.100
—
0.061
0.070
0.01

1.028 t 0.646
0.2909 ± 0.1606
0.789 t 0.649
0.132 t 0.011
1.2
0.111 ± 0.018
0.231 t 0.056
0.12 ± 0.01
0.834 ± 0.560
0.313 ± 0.130
0.104 ± 0.009
0.159 ± 0.075
1.545 ± 1.538
0.988 ± 0.0097
0.111 ± 0.027
0.141 ± 0.117
0.37 ± 0.05
(24-hour)
Singh et al.
Ibid.
Singh, et al
Ibid.
Cronn, et al
Ibid.
Ibid.
Cronn, et al
Ibid.
Ibid.
Ibid.
Ibid.
Ibid.
Singh, et al
Singh, et al
Ibid.
Simmonds et

, 1981

. . 1979a

. , 1979


. , 1977a





.. 1978c
. , 1979a

a)., 197-

                                                                          0.10
                                                                    Lillian et al.,  1975

-------
                                                              •  TABLE  3-7.   (continued)
oo
 I
Location Type of site
Hawai i
Cape Kumakahi Remote
Ireland
Western Ireland Remote
Japan
Tokyo Urban
Tokyo
Kansas
Jetmar Remote
Louisiana
Baton Rouge, Urban
Geisinar, Plaquemine
Maryland
Baltimore Urban
New Jersey
Seagirt Marine
Sandy Hook Marine
Bayonne Urban
Pathway/ Urban
Woodbridge,
Boundbrook, Passaic
New York
New York City Urban
Niagara/Falls/ Urban
Buffalo
White Face High altitude
Mountains
Ohio
Wilmington Urban
Oregon
Cape Meares Remote
Panama Remote tropo-
sphere
Maximum
Date (ppb)

Nov. 1979 - Jan. 1981

June/July 1974

9/10 - 10/27, 1975 20.60
May 1974-Apr 1974

6/1-6/7, 1978 0.159

Jan/March 1977 1.6


7/11-7/12, 1974 0.21

6/18-6/19, 1974 0.20
7/2-7/5, 1974 0.33
March-December, 1973 14.4
Sept. 1978 20.5



6/27-6/28, 1974 1.6
July 1978 1.0

9/16-9/19, 1974 0.13

7/16-7/26, 1974 0.35
Jan. 1979 - Jan. 1981
7/77

Minimum Average
(ppb) (ppb)

~ 0.14

0.048 t 0.0172

0.20 1.70 ± 1.70
0.800

0.106 0.130 ± 0.016

trace 0.11


0.044 0.12

0.044 0.10
0.030 0.15
0.075 1.59
trace 11.4



0.10 0.61
0.26 0.66

0.032 0.067

0.030 0.097
~ 0.155
0.97 ± 0.05

Reference

Rasmussen

Lovelock,

Tada et al
Ohta et al

Singh, et

Pellizzari


Singh, et

Lillian et
Ibid.
Ibid.
Pellizzari



Lillian et
Pellizzari

Lillian et

Ibid.
Rasmussen
Cronn and
1979


and Khalil, 19816

1974

. , 1976
. . 1977

al., 1979a

et al., 1979


al.. 1979a

al., 1975


et al., 1979



al., 1975
et al., 1979

al., 1975


and Khalil, 1981b
Robinson,

                Samoa  (14°S)
Remote
                                                  Feb. 1980 - Jan. 1981
                                                                                                   ~ 0.155
                                                                                                                     Rasmussen and Khalil,  1981b

-------
TABLE 3-7.   (continued)

Location Type of site
Soviet Union
Caucasus
Mountains
Tasmania
Cape Grin

Tennessee
Smoky
Mountains
to
w Texas

Houston,
Deer Park,
Pasadena
Northern Mid-
Latitudes
Washington
Pull nan
Pullman
Pullman
Pullman
Pullman
Pullman

Pullman
Pul Iman

Spokane

Remote


Remote


Regionally
polluted



Urban


Remote tropo-
sphere

Remote
Remote
Remote
Remote
Remote
Remote

Remote
Remote tropo-
sphere
Urban
Maximum
Date (ppb)

7/79 0. 12


Oct. 1976 - March 1977
Jan. 1979 - June 1980

9/78 0.32




July 1976 5.1


6/76 0. 11


1/75
1/76
1/77
10/77 - 11/77
1/78
1/79

1/80
3/76

4/78
Minimum Average
(ppb) (ppb)

0.15 0.13 ± 0.06


0.012 ± 0.003
- 0.1

0.11 0.19 t 0.03




trace 0.44


0.07 0.095 t 13


0.090
0.098
0.109
0.115
0.120
0.135

0.157
0. 095 i 0. 008

0.5
Reference

Cronn, 1980b

,
Fraser and Pearman,
Rasmussen and Khali 1

Cronn and Harsch
1979b



Pellizzari, et al.,


Robinson and Harsch,
1978

Robinson, 1978
Ibid
Ibid
Ibid
Ibid





1978
. 19816






1979










Khali] and Rasmussen,
1980
Ibid
Cronn et al., 1976

Cronn et al., 1979






-------
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Noweir, M. H. ,  E.  A.  Pfitzer, and T. F. Hatch.   Decomposition of chlorinated
     hydrocarbons:   a  review,  J.  Amer.  Ind.  Hyg. Assoc. 3_3(7): 454-460, 1972.

Ohta, T., M.  Morita,  I.  Mizoguchi,  and T. Tada.  Washout  effect and diurnal
     variation for chlorinated hydrocarbons  in  ambient air.  Atmos. Environ.
     11:985-987, 1977.

Pellizzari, E. D.  Electron capture  detection in gas chromatography. J. Chromat.
     98:323-361, 1974.

Pellizzari, E. D.  Measurement of  Carcinogenic  Vapors  in Ambient Atmospheres.
     EPA-600/7-78-062, 1978.

Pellizzari, E. D. and J.  E. Bunch.    Ambient Air Carcinogenic Vapors:   Improved
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     1979.

Pellizzari, E.  D. ,  M.  D.  Erickson,  and R.  A.  Zweidinger.   Formulation of  a
     Preliminary Assessment of Halogenated Organic Compounds  in  Man  and Environ-
     mental Media.  EPA-560/13-79-006, 1979.

Pierotti, D., R.  A.  Rasmussen, and  R. Dalluge.   Measurements of N20,  CF2C12,
     CFC13, CH3CC13, CC14, and CH3C1  in the troposphere and lower stratosphere
     over North America.   J. Geomagn. Geoelect.  32:1-2, 1980.

Piet, G.  J.,  P.  Slingerland,  F. E.  de Grunt, M. P.  M. v.d. Heuvel,  and B.  C.
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Rasmussen, R. A.  and M.  A.  K.  Khalil.   Global  atmospheric distribution  and
     trend of methyl chloroform (CH-CCK).  Geophy.  Res. Lett. 8 (9):
     1005-1007, 1981b.             J   J

Rasmussen, R. A.  and M.  A.  K.  Khalil.   Interlaboratory comparison of fluro-
     carbons  -11,  -12, methyl  chloroform and  nitrous oxide  measurements.
     Atmos. Environ. 15:1559-1568, 1981a.

Rasmussen, R. A.,  D.  E.  Harsch, P.  H. Sweany, J.  P.  Krasnec,  and D. R. Cronn.
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     chromatographic  freezeout concentration method.   J.  Air Poll.  Control
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Robinson,  E.  Analysis of Halocarbons in  Antarctica.  WSU report 78/13-42  for
     the  National Science  Foundation, December,  1978.

Robinson,  E., and D.  E.  Harsch.   A  Halcarbon Survey in the Pacific Area  From
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     Association, June, 1978.

Russell,  J.  W. , and L. A.  Shadoff.   The sampling and determination of halo-
     carbons  in  ambient  air  using   concentration on  porous  polymer.   J.
     Chromat. 134:275-284,  1977.


002MC4/B                            3-40                               3/25/82

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Sheldon, L. S.,  and  R.  A.  Hites.  Organic  Compounds in the Delaware River.
     Environ.  Sci. Technol. 12(10):1188-1194, 1978.

Simmonds, P. G.,  S.  L.  Karrin, J. E. Lovelock, and F. H. Shair. Distribution
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Singh, H. B.   Atmospheric  halocarbons:   Evidence  in favor of reduced average
     hydroxyl  radical concentration  in  the troposphere.  Geophy.  Res. Lett.
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Singh, H. B.,  L.  J.  Salas, and  R.  Stiles.   Trace Chemicals  in  the  "Clean"
     Troposphere.  EPA-600/3-81-055, October 1981.

Singh, H. B.  Preliminary estimation of average tropospheric HO concentrations
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Singh, H. B.,  L.  J.  Salas, and  L.  A.  Cavanagh.   Distribution,  sources  and
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Singh, H. B.,  L.  J.  Salas, D. Lillian, R. R. Arnts, and A. Appleby.  Genera-
     tion of  accurate  halocarbon primary standards with permeation tubes.
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Singh,  H.  B. ,  L. J.   Salas, H.  Shigeishi, and A.   Crawford.   Urban-nonurban
     relationships of halocarbons, SF6, N20 and other atmospheric trace consti-
     tuents.  Atmos.  Environ.  11:819-828, 1977c.

Singh, H. B.,  L. J. Salas, H,  Shigeishi, and E. Scribner.  Global Distribution
     of  Selected Halocarbons, SF6  and  N20.   Phase  II  Interim Report, SRI
     International, Menlo Park, California, May 1978a.

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     of  Selected  Halocarbons, Hydrocarbons,  SF6,  and N20.   SRI  Intern.,  Menlo
     Pk., CA,  EPA-600/3-78-100.   U.S. Environmental Protection Agency, Research
     Triangle Park, NC, December  1978b.

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     Park, CA, EPA-600/3-79-107,  U.S. Environmental  Protection Agency, November
     1979a.

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     California, October 1979b.
002MC4/B                            3-41                               3/25/82

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Singh, H. B.,  L.  J.  Salas, A. J.  Smith,  and H. Shigeishi.  Measurements of
     some potentially  hazardous organic  chemicals in  urban environments.
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World Meteorological Organization  (WMO).   Global Ozone  Research  and Monitoring
     Project,  Report No.  11.  The  Stratosphere  1981:  Theory and Measurements,
     World Meteorological Organization, Geneva,  Switzerland, 1982.

Wuebbles, D.  J.   The  relative efficiency  of a number of halocarbons for des-
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     1981.
 002MC4/B                             3-42                               3/25/82

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                      4.   METABOLIC FATE  AND DISPOSITION
4.1  ABSORPTION, DISTRIBUTION, AND ELIMINATION



     Methyl chloroform  (MC;  1,1,1-trichloroethane)  is currently one of  the



most widely used of the chlorinated aliphatic hydrocarbon solvents. Since the



introduction  in  the  mid-19501s  as  a cold cleaning solvent  substitute  for



carbon tetrachloride,  MC has gained recognition as being among the  least toxic



of the chlorinated aliphatic  hydrocarbons and is  increasingly replacing other



supposedly more toxic  chlorinated solvents such as trichloroethylene  (Torkelson



et al., 1958;  Prendergast  et al., 1967; Stewart, 1968).   The fact that its



isomer, 1,1,2-trichloroethane,  is markedly  more toxic than MC (Hardie,  1964;



Irish, 1963;  Browning, 1965;  CaVlson, 1973),  is in agreement with the general



observation that chlorinated ethanes having chlorines  on both carbon  atoms  are



considerably  more  soluble  in water,  blood, and  lipid  (Table  4-1).   These



physical  properties are responsible  in  part for the manifestations of chlori-



nated hydrocarbon toxicity (Sato and Nakajima, 1979;  Clark and Tinston,  1973).



4.1.1  Oral and Dermal Absorption



     While  limited absorption of MC vapor  through the  lungs is the common



route of entry  into the body, MC  is  also  rapidly  and  completely absorbed from



the gastrointestinal  tract  (Stewart,  1971).   Stewart and Andrews  (1966) re-



ported an  instance of non-fatal acute  intoxication after oral ingestion of a



liquid ounce  of MC (0.6 g/kg  body weight,BW).   The concentration of MC  in the



expired air was measured serially and found equivalent to an inhalation exposure







002MC2/B                             4-1                              11-10-81

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TABLE 4-1.  PARTITION COEFFICIENTS OF METHYL CHLOROFORM AND OTHER SOLVENTS AT 37°C

Vapor Pre
Compound torr at 2
1,1,1-Trichloroethane
1,1,2-Trichloroethane
1,1-Dichloroethane
1,2-Dichloroethane
Tri chl oroethy 1 ene
Tetrachl oroethyl ene
Dichloromethane
Chloroform
Carbon tetrachloride
125
25
250
80
436
20
400
250
100
iss Water
'5C Air
0.93
17.1
2.7
11.3
1.3
0.43
7.6
3.5
0.25
Olive Oi
Air
356
2273
187
447
718
1917
152
401
361
1 Blood
Air
3.3
38.6
4.7
19.5
9.5
13.1
9.7
10.3
2.4
Olive Oil
Blood
108
59
40
23
76
146
16
39
150
Adapted  from Sato and Nakajima, 1979.
 002MC2/B                             4-2                              11-10-81

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of experimental subjects  to  500 ppm.   On the other hand, MC vapor is poorly
absorbed through  intact  skin and,  unless it is trapped against the skin be-
neath an  impermeable  barrier, it is unlikely that  toxic  quantities  can be
absorbed  (Stewart,  1971;  Stewart and Dodd,  1964).  Stewart  and Oodd (1964)
demonstrated, with  continuous  immersion  for 30  min of the thumbs  or  hands  of
volunteers, that  the  solvent penetrates  the skin, enters the blood circula-
tion, and  is  excreted through the lungs in  exhaled  air.   Fukabori  and co-
workers (1976,  1977)  attempted  a  quantisation  of  cutaneous  absorption in
humans.   Systemic absorption was evaluated by measurement of MC in  blood and
exhaled air and its biometabolites [trichloroethanol (TCE) and trichloroacetic
acid (TCA)}  in  urine.   After applications  of the solvent to the skin of the
                                         o
forearm in a  circumscribed  area (12.5 cm)  for 2 hr a day for 5 consecutive
days, or  immersion  of the hands 11 times  a dayfor 10 min periods,  the MC
concentrations  in exhaled air and urinary metabolites corresponded roughly to
a 2-hr  inhalation exposure to  10-20 ppm MC  in ambient air.  The investigators
concluded that  absorption through the skin  for workers in direct contact with
liquid MC may significantly  (5%) add to the absorption from vapor exposure.
4.1.2  Pulmonary Uptake and Body Burden
     Possibly because of  its reputation  as an organic solvent with  low human
toxicity,  few  studies  have  been made of the pharmacokinetics of MC  pulmonary
uptake,  concentration in the body, and metabolism after exposure to  low  inha-
lation  concentrations approximating  the accepted  TLV-TWA concentration.
Information available from these studies  relate  to single exposures  in experi-
mentally controlled conditions with  animals or human  volunteers.   Little
information is  available  regarding  long-term exposure,  such as may occur  in
002MC2/B                             4-3                              11-10-81

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the workplace or in the environment.  Recent studies have greater reliability

than earlier studies because of the availability of  gas  chromatographic methods

for the analytic determination of MC and its metabolites in  alveolar  air, body

fluids, and tissues (Monster et al., 1979b;  Monster  and  Boersma,  1975; Astrand

et al.,  1973;  Eben and Kimmerle,  1974;  Briemer et  al., 1974; Humbert and

Fernandez,  1976,  1977;  Stewart  et  al.,  1969),  although spectrophotometric

methods based on adaptations of the Fujiwara reaction are still widely used  by

many  investigators  for  determining  urinary excretions  of trichloroethanol

(TCE), trichloroacetic acid (TCA),  or total urine  chloro-derivatives of MC

(Ogata et  al.,  1974;  Imamura and Ikeda, 1973; Tanaka and Ikeda,  1966; Ikeda

and Ohtsuji, 1972).

     Inhaled MC  rapidly equilibrates with arterial  capillary blood across the

lung  alveolar  endothelium (Astrand, et  al., 1973).   The rate  of pulmonary

uptake or absorption depends largely on the solubility of MC in blood (Ostwald

solubility  coefficient) and hence the blood/air partition coefficient  (Table
                                *.
4-1).  Vapors with a  high partition coefficient are  absorbed  into  the  body

readily and achieves  a high level  of accumulation.   Conversely,  vapors with  a

low partition coefficient accumulate slowly because and, partition occurs more

rapidly with venous  blood and alveolar air, are more rapidly eliminated into

expired air following exposure.   In comparison with other common solvents in

Table  4-1,  MC  possesses  a  relatively  low blood/air partition  coefficient of

about  3.3 at 37°C.

     The  magnitude of MC  uptake into the  body  (dose,  burden) is directly

related to  the following factors:   1) concentration of MC in the inspired air;

2) duration of  exposure;  3) pulmonary ventilation  during exposure;  4) blood/

air  partition  coefficient;  5)  rates of diffusion into,  and  solubility  in,  the
002MC2/B                             4-4                               11-10-81

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body tissues; and 6) total body-lipid repository.   Consequently, during expo-



sure at a given inspired air concentration, pulmonary uptake and retention is



initially large and gradually  decreases  to a minimum steady-state value  as



total body equilibrium  (and  body  burden) with inspired  air concentration,  is



reached.   At this  steady-state condition, pulmonary uptake  balances the pulmo-



nary and other routes  of  elimination,  including metabolism.  For any  given



breath cycle during exposure,  the proportion of MC absorbed and retained by



the body is equal  to the inspired air concentration (C,) minus  the end  alveolar



air concentration (C.)  and,  since pulmonary uptake is  a function of  inspired



air concentration, the percent retention is:


                                                   r  -  r
                                                    I     A
     % retention of inspired air concentration = 	   x 100     (1)

                                                     CI


This value is large at the beginning of exposure,  but  gradually decreases  to a



constant minimum  value  as  total  body equilibrium is approached (Figure 4-1).



The percent  retention value  at equilibrium is  independent  of the inspired air



concentration at equilibrium.  Curing experimental exposures of subjects to 70



and 140 ppm  MC  for 4 and  8  hr,  both Monster et al. (1979b) and Humbert and



Fernandez (1977)  found  retention  to be 30 percent of  inspired air  concentra-



tion at an equilibrium reached after 4 hr of exposure.


     The total amount  (dose, Q)  of MC retained in the body during  an inhala-



tion exposure can be estimated by  multiplying  percent retention  by the  volume



of air inspired during the exposure period, or:



                  Q = (Cj  - CA)  V  • T                                (2)


where V  is ventilation  rate  (£/min)  and  T  is  exposure period (min).  Since  the



retention value decreases  exponentially  during the  exposure until equilibrium



is  reached  (Figure 4-1),  either  experimental  measurements of retention  at



frequent intervals during exposure (Monster et al., 1979b)  or integration over







002MC2/B                             4-5                              11-10-81

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r
o
EC
Z CC

O a
u 8j

O 2
o
LLJ
>
-J
<
100
      so
EXPOSURE
 I   I    I   I   I


POST EXPOSURE • MC
                                                       I    1   I
                                                 BLOOD CONC.
         __     "'I	^	^^i
                                       10



                                    TIME, hours
                                                           20
   Figure 4-1.  Absorption and pulmonary elimination of MC, and blood

   concentration (see text for explanation).



   Source: Davidson (1980).  t
                                  4-6

-------
the  experimentally  determined retention  function  is  required (Humbert and


Fernandez, 1977).  Ifce  exponential  nature of the retention curve  is  due  to


first-order kinetics of saturation of body compartments with MC.


     Table 4-2  shows  the  amounts of MC absorbed into the body during inhala-


tion exposures  for  volunteers from the studies of Monster et al.  (1979b)  and


Humbert and Fernandez  (1977).   Monster and his coworkers estimated pulmonary


uptake  of  MC  by multiplying  the minute  volume  by percent retention during


single  4-hr exposures  to  70 ppm and 142 ppm.  They observed a direct propor-


tionality  between  uptake  and  inspired air  concentrations  of MC.   Similar


results were obtained by Humbert and Fernandez for volunteers exposed for  8 hr


to  72  ppm and  213 ppm.   Comparison of pulmonary uptake  for  4-hr  and  8-hr


(Table 4-2) exposures indicates that the amount of MC retained is also propor-


tional  to  duration  of exposure,  although the values for uptake determined by


Humbert and Fernandez  are 40 percent lower than those of Monster et al.  This


variation can be ascribed to differences in the experimental methodologies and
                               t

average minute  volume.  Nonetheless, these experimental  results indicate that


the body burden resulting from an 8-hr inhalation exposure to 350 ppm (TLV-TWA)


approximates 1.5 to 2 g of MC for a normal 70-kg man.


     The body  uptake  of MC  increases with the duration of  inhalation exposure


and with physical work  or exercise.  Monster et  al. (1979b) found  that during


a 4-hr  exposure to  142 ppm MC, with physical  activity equivalent to  light


physical work  (100 watts),  pulmonary uptake increased from 429 mg to  538 mg


MC, an increase of 25 percent (Table 4-2).  This increase was primarily due to


an  increase  of ventilation from 10.7  fmin    sedentary to 30.6 fmin   with


work.  While  it might be  expected  from equation  (2) that  the  increased uptake


should be directly proportional to the ventilation rate increment, an increase
002MC2/B                             4-7                              11-10-81

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 TABLE  4-2.   ESTIMATED UPTAKE  OF  MC  DURING A SINGLE  4-HR  INHALATION  EXPOSURE
   (AVERAGE  BODY WEIGHT,  77 kg; 67  kg LEAN BODY  MASS),  MONSTER  ET  AL.,  1979

Subject


A
B
C
D
E
F
Av.
Exposure
70 ppm (at rest*)
Uptake, mg
140
240
200
185
200
190
193
Concentration
145 ppm

305
520
465
395
425
465
429

142 ppm*, with work (100 watt)
Uptake, mg
435
610
540
560
575
505
138
"•Ventilation minute volume increased from average of 10.7 £/min. at rest to
30.6 je/min during light work.
        ESTIMATED UPTAKE OF MC DURING A SINGLE 8-HR INHALATION EXPOSURE
           (AVERAGE BODY WEIGHT-74 kg), HUMBERT AND FERNANDEZ, 1977

Subject


BH
JC
JO
Av.

72

293
274
277
281
Exposure Concentration*
ppm 213 ppm
Uptake, mg
921
912
—
917
*Average ventilation minute volume 5.7 £/min.
 002MC2/B                              4-8                              11-10-81

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in ventilation also tends  to increase alveolar elimination  of MC  from pulmo-



nary venous blood. Similar observations  were made by Astrand et  al. (1973)



with volunteers exposed  to an MC inhalation concentration of 250  and 350  ppm



for 30 min at rest, alternating with  identical exposure plus 50 watt of  light



work.   Table  4-3  shows  that  their  subjects responded to this work with  a



3-fold increase of pulmonary  ventilation, nearly a 2-fold increase in cardiac



output, and a 50 percent increase in arterial blood concentration, which is an



index of pulmonary uptake.   Also,  end alveolar concentration of MC increased



while retention decreased from 50 to 33 percent with work.   Therefore, physical



activity during MC exposure increases the  uptake  and  body  burden, but  the



increase is not directly proportional to increased ventilation and is self-



limited by a  compensatory increase of pulmonary  elimination.  For solvents



like MC, with  a  low  blood/air partition coefficient, physical  activity  has a



smaller effect on net pulmonary  uptake and  body retention than those solvents



with a high blood/air partition coefficient (e.g., trichloroethylene, perchloro-



ethylene, dichloromethane; Tablt? 4-1; Monster, 1979).



     During inhalation of MC, and in the elimination phase after exposure, the



arterial blood concentration  always is directly proportional  to the alveolar



concentration (Monster  et  al., 1979b;  Humbert  and  Fernandez, 1977; Astrand et



al., 1973; Gamberale and Hultengren, 1973; Eben and Kimmerle,  1974).   Since at



equilibrium the end alveolar air concentration is proportional  to  inspired air



concentration, blood  concentration of  MC  is  also  related  to  inspired  air



concentration (Monster  et  al.,  1979b; Number and Fernandez, 1977; Astrand et



al., 1973).   This  fixed relationship between  alveolar air  and blood concen-



tration is defined by the  blood/air partition  coefficient for MC.   Figure 4-2



illustrates this  linear relationship for a  volunteer  exposed  for 30  min to
002MC2/B                             4-9                              11-10-81

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              TABLE  4-3.   MEAN  VALUES  AND  SEM  FOR  12 MALE  SUBJECTS
                  AT REST  AND EXERCISE FOR 30  MINUTE PERIODS


MC
At rest
250 ppm
350 ppm
50 Watt
250 ppm
350 ppm
Ventilation
BTPS
i/min
6.6 ± 0.4
6.6 ± 0.4
22.5 ± 1.0
21.8 ± 1.1
Cardiac Alveolar
output cone
£/min ppm
5.1+0.4 125 ± 6
179 ± 13
9.7 ± 0.6 168 ± 7
239 ± 17
Blood
arterial
cone.
M9/9
3.0 ± 0.
5.0 ± 0.
4.5 ± 0.
7.2 ± 0.


2
5
2
4
From Astrand et al.  (1973).
002MC2/B
4-10
11-10-81

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 I
 o
 cr
 z
 w
 o
 o
 u
 cc
 w
                              I
                                        J_
                  100        200       300        400


                  ALVEOLAR CONCENTRATION, ppm
Figure 4-2.  Relationship between methyl chloroform concentration
in alveolar air and arterial blood. Data from one subject. Product
moment correlation: r = 0.985.  Data from  one subject exposed to
30-min. periods of MC.  Alveolar air samples and arterial blood sam-
ples.

Source:  Gamberale and Hultengren (1973).
                                 4-11

-------
increasing increments of MC  concentration  in inhaled air.  The relationship
was  independent  of  the  duration of exposure over  the  time  period studied.
Eben and Kimmerle (1974) exposed  rats to 204 ppm for 8 hr daily,  5 days  a wk
for 14 weeks  and found  that the blood concentration of MC determined immedi-
ately after daily exposure  remained constant during the entire 3 mo period.
     The blood/air partition coefficient, as determined from in vivo measures
of alveolar air  concentration and blood concentration of MC, agrees well with
the jm  vitro  value  of 3.3  at  37°c  determined by Sato  and  Nakajima  (1979)
(Table 4-1).  Table 4-3  summarizes  the data of Astrand et al. (1973) deter-
mined for  sedentary men exposed to 250  and 350 ppm MC for  30-min periods
alternating with 30-min periods of physical  activity.  The ratios of arterial
blood concentration  (ug/g)  to alveolar MC concentration remain nearly constant
over a 2-fold range  of  alveolar concentrations, with an average value of 5.
Monster et al. (1979b) estimated from their study of men exposed to 70 and  142
ppm MC a blood/alveolar air concentration ratio of  6.   In comparison to other
solvents (Table  4-1), MC has a relatively small  blood/air partition coefficient,
and hence for equivalent ambient air exposure concentrations the blood concen-
tration of MC is proportionally lower than for other solvents.
     The amount  of MC pulmonary uptake is influenced by total body weight and
also by the total  fat content of the  body  (average body fat = 8 percent of
body weight).   The  capacity of adipose tissue  to  absorb  MC ni vivo  is de-
termined by the  product  of adipose tissue volume and the lipid solubility  of
MC.  The lipid/blood partition coefficient for MC (108 at 37°C) is higher than
for most other  structurally related solvents (Table 4-1) and, therefore, the
capacity of adipose tissue  for MC is  relatively high.  However, because of the
low rate of perfusion (5 percent cardiac output),  the  time  needed  to  saturate
002MC2/B                             4-12                             11-10-81

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adipose tissue  is  large in comparison with that  for  other tissues.   Also,

since the blood/air  partition  coefficient  for MC  is lower than that  of  other

solvents,  the amount of MC in adipose tissue  (concentration) at the end of an

exposure of similar duration will be relatively lower.   Monster (1979)  suggests

that because of the  high solubility of MC in  adipose tissue, accumulation may

occur with  repeated daily exposures, particularly  in  obese persons.  This

speculation is  supported by  the observations  of Savolainen, Vainio  and co-

workers (1977,  1978),  who  exposed rats  to MC  (500 ppm) 6 hr daily for  5 days

and determined MC concentrations in perirenal  adipose tissue and in other body

tissues.  Their data, shown in Table 4-4, indicate that measureable amounts of

MC remained in  perirenal  fat tissue 18 hr after the previous exposure  of day

4, and  markedly increased further with a 6-hr exposure on day 5.  The adipose

tissue/blood partition  coefficient calculated  from these data was  21,  as

compared to 1.6  for brain  and  liver tissues.   On  the other hand, Eben and

Kimmerle (1974) exposed  rats  for 14 weeks (8 hr daily,  5 day per wk) to 204
                                «.
ppm MC but failed to find MC in*adipose or other tissues and concluded  that MC

did not accumulate with chronic exposure. However, the average exposure concen-

tration for their  rats was lower  than that used by  Savolainen et al. (500 ppm

vs. 204 ppm).

4.1.3  Tissue Distribution

     During exposure to  MC,  distribution of the compound throughout the body

and the amount  concentrated by  each tissue is primarily  governed by  the blood

concentration,  the blood perfusion rate,  and  the affinity of MC for respective

tissues, as determined by individual tissue/ blood partition coefficients.   In

comparison with other  chlorinated hydrocarbon solvents  that are known  to be

distributed widely  in  the  body, MC has  one of  the  highest  lipid/ blood par-

tition  coefficients  (108 at  37°C, Table 4-1) and would  distribute into all



002MC2/8                             4-13                             11-10-81

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         TABLE 4-4.   TISSUE CONTENT (RAT)  OF  METHYL  CHLOROFORM  (MC) AFTER CHRONIC  INHALATION EXPOSURE OF 500 ppm
   Exposure on
    5th  day
       hr.                  Cerebrum          Cerebellum            Liver          Perirenal fat         Blood


                                      nmolMC/g wet weight  +  range

        0                 0.15 +  .03        0.17+   .03     0.15 +   .01       16.9+     .5      0.08+   .01

*.       2                146  +1.1         14.0  +   2.2       14.7  +   .04      183.5+  10.7     11.5  +  2.0
M
*       3                13.4  +  .60        13.2  +   .8       15.7  +3.3       218.9+  63.4      8.5  +  1.0

        4                12.2  +  .40        15.9  +   1.5       16.2  +2.8       261.2+  19.1     12.7  +  2.9

        6                15.6  +4.6         21.3  +   9.6       21.3  +   .4       276.0+  30.1     13.1  +  1.9



    Measurements were performed on the 5th  day of exposure after 4 previous daily exposures, 6 hr daily.

   From Savolainen et al.  (1977).

-------
body  tissues,  particularly  those high in lipid content,  such  as  brain and

adipose tissue (Holmberg et al., 1977). Table 4-4 shows the concentrations of

MC In  liver,  brain,  and adipose tissue of  rats exposed  to 500 ppm in their

inspired air  for  6  hr daily for 5  days  (Savolainen  et al., 1977).   Adipose

tissue appears to have a partition coefficient with blood of approximately 20,

and brain tissue levels are also greater than blood concentration.   In man, MC

readily passes the  blood-brain  barrier,  resulting in  high  concentrations in

the brain (Caplan et al., 1976) and in the cerebrospinal  fluid (Larsby et al.,

1978).  While  it  has not been demonstrated  directly  to  cross  the placenta!

barrier into  the  fetus,  it may be expected  to do  so (Laseter and Dowty, 1977)

like  other  highly lipid-soluble  haloalkanes.  Trichloroethanol  (TCE),  a major

metabolite  of  MC, is known to cross readily into the fetus (Bernstine, 1954,

1957).  While  there is no report  of  MC occurring in  colostrum or milk of

nursing mothers,  it  may be expected to distribute readily into  these  compart-

ments because of their high lipid content.
                               *
4.1.4  Pulmonary Elimination   »

      Figure 4-1  shows  schematically the time-course of pulmonary elimination

of MC after exposure.  At termination of exposure, MC  immediately begins to be

eliminated  from  the  body  into  the  lungs with blood concentration and  alveolar

air  concentration  describing parallel  exponential  decay curves with  three

major  components.   These  components represent first-order passive diffusions

of MC  from  three major body compartments:   (1) most rapidly from a vessel-rich

group  of  tissues  (VRG) with high blood flow and  high  diffusion rate constant

(VRG: brain, heart,   kidneys, liver, endocrine, and digestive system),  (2) more

slowly  from the  lean body mass  (MG;  muscle and skin) and  (3)  from  adipose

tissue  (FG) (Fiserova-Bergerova  and Holaday, 1979).   The rate constants  for
002MC2/B                             4-15                             11-10-81

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the passive diffusion  from  VRG,  MG and FG compartments  are  dependent  on  both



the arterial blood flow/tissue mass and the relative solubilities  of HC  in  the



tissues of these compartments (tissue/ blood partition coefficients).  However,



the ranking of  half-times (t,,-) of elimination of MC is VRG < MG < 
-------
(1979b) and Humbert and Fernandez (1977) observed that, after single controlled



inhalation exposures, pulmonary  elimination was not complete for 6 to 10 days



as determined  by  the  continued presence of MC  in  alveolar air.   The possi-



bility  of  significant accumulation  with  repeated daily  exposures  to this



solvent  (Laseter  and Oowty,  1977),  however,  is offset in part by  the  low



blood/air partition coefficient  (Table  4-1),  which for a given exposure con-



centration limits pulmonary absorption and body dose.



4.1.5  Elimination by Other Routes



     There is  no  report  in  the literature of significant  elimination of MC by



any route other  than  pulmonary.   MC is poorly  soluble in water,  even when



compared with  other  chlorinated  hydrocarbons  (Table 4-1), and with  its high



lipid/water partition coefficient MC is unlikely to be excreted unchanged in



the urine in any significant amounts.  Studies of chlorinated compounds  in the



urine after exposure of animals and humans to MC have not reported its appear-



ance in  urine  (Monster  et al.,  1979; Eben and  Kimmerle,  1974; Humbert and



Fernandez, 1977;  Seki et  al.,  J975;  Stewart et  al., 1961;  Hake et al., 1960).



Since highly lipid soluble substances like MC readily cross into the intestinal



lumen,  some  fecal and  flatus excretion can be expected  during  inhalation



exposures.    After controlled  inhalation  exposures,  Humbert and  Fernandez



(1977) were  able  to  account for 88  to  100 percent of an estimated retained



dose as  unchanged  MC  in postexposure exhaled air and as metabolites of  MC in



urine.   However,  Monster et al. (1979b)  could account for only 60  to 80  percent;



furthermore,  they  noted  that  the percentage  recovered decreased with higher



exposure doses.



4.2  BIOTRANSFORHATION



     MC has long been known to be metabolized to only a very limited extent by



mammals.  The  generally accepted metabolites of MC —  trichloroethanol (TCE),







002MC2/B                             4-17                              11-10-81

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TCE-glucuronide, and trichloroacetic acid  (TCA)  •-  are excreted primarily  by



the kidney, but  very  small  amounts of TCE  (<1 percent)  are  excreted by the



lungs (Monster et al.,  1979b).   TCE-glucuronide is also excreted to an unknown



extent in bile  (Owens  and Marshall, 1955).  No  other  metabolites  have been



reported.



4.2.1  Magnitude of MC Metabolism



     Only one balance study with isotopically labeled MC has  ever been carried



out.  More than 20 years ago, Hake and his coworkers (1960),  using   C-labeled



MC, determined that  less  than  3 percent of MC is metabolized by rats.  More



recently, estimates of  the  extent of metabolism  in  man  have been made from



controlled inhalation exposures with unlabeled MC (Seki et al., 1975; Monster



et  al.,  1979b; Humbert  and  Fernandez, 1977).   From  the experimentally  deter-



mined retained dose and the amounts of MC metabolites excreted into the urine,



the percent of the dose metabolized in man is estimated to be about 6 percent.



     Hake' et  al.  (1960) found that  98.7  percent  of a dose of MC given to rats

                               »•.                                        14
was eliminated  unchanged  via the lungs.   These  investigators  injected   C-


                                14
labeled  1,1,1-trichloroethane-l-  C  (700 mg/kg)  intraperitoneally  into  3 rats



(170-183 g),  which  were then placed individually in a Roth  metabolism unit



with air traps  for  14C-MC (cold  toluene),  14CO«  (NaOH  solution) and finally a



trap for "metabolites"  (quartz-tube furnace and halogen  absorber that con-



verted any halogenated  hydrocarbon  to inorganic  halogen  and  C0_,  and absorbed



the halogen).   Each rat was kept in the unit for 25 hours, during which time



urine and feces were collected.   At necropsy at  25 hr, blood and tissue samples


                        14                  14
were analyzed for total    C-activity.   The  C-MC used in the  experiments  was



synthesized by the  investigators and was determined  by paper chromatography to



be  99 percent pure, with  possible contamination  by 0.4 percent 1,1,2-trichloro-



ethane isomer and 0.5  percent 1,1-dichloroethane.   Their findings, summarized






002MC2/B                            4-18                              11-10-81

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in Table 4-5,  Indicate that metabolism of MC in the rat is limited to about 2
percent of a very large dose.   However,  only TCE-glucuronide  was identified as
                                                  14
a metabolite  in  the urine, and the source of the   C02 in expired air could
                                                            14
not be defined because  of the presence of small amounts of   C-l,l,2-isomer
    14
and   C-dichloroethane in the dose.
     Man also appears to possess a very limited capacity for  metabolism of MC.
Table 4-6 summarizes data taken from several investigative studies showing the
amount of urinary  metabolites excreted  after exposure to various concentra-
tions (4.3  to 213  ppm)  of MC  in  inspired  air.   Seki  et al.  (1975) surveyed
workers in  4  printing plants  where MC was  the  sole organic  solvent in use.
The workers were exposed 8 hr daily, 5 1/2 days per wk, over a period of  at
4
least 5 years.   Urine samples were collected in the latter half  of the work
week.   Their  data  in Table 4-6 show a proportional increase of both TCE  (as
glucuronide)  and  TCA, and  also  total  trichlorinated  compounds  (TTC)  with
increased ambient  air concentrations of MC.  Seki et al. expressed their data
as a linear relationship between inspired MC concentration and urinary metabo-
lite excretion, with an  added constant increment presumably due to continued
excretion from the  previous  day's exposure.  From  this observation and from
evidence of continued excretion during exposure-free weekends, they suggested
that MC accumulates in  the body to a steady-state body burden defined by the
air concentration  of each daily exposure.   Seki et al.  also noted that the
amounts of  metabolites excreted daily in the urine  of  these workers were  less
than 5 percent of that observed with comparable exposures to trichloroethylene,
of which 60 to 80  percent is metabolically degraded to TCE and TCA (Monster,
1979).   This  comparison  suggests  that  about 5 percent or  less MC is metabo-
lized.
002MC2/B                             4-19                             11-10-81

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        TABLE 4-5.   HAKE ET AL.  (1960) RECOVERY EXPERIMENT  WITH  RATS  (3)
              INTRAPERITONEALLY  INJECTED WITH J«C-MC  (700 »g/kg)
                                                  Average % dose


Expired Air

     Unchanged MC (by isotopic dilution)                 97.6

     Unknown compound detected by
       furnace (assumed to be unchanged MC)               1.1

     14C02 (in NaOH) 70% within 4 hr; 100% within 12 hr   0.5
Urine
     TCE-glucuronide •+• other volatile compounds but
     no detectable TCA                                    0.85
Feces

     Uncharacterized           -                          0.03
                               %


Tissues

     Uncharacterized except skin (90% unchanged MC)       0.18
002MC2/B                             4-20                              11-10-81

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     The data (Table 4-6) of Monster et al.  (1979b)  and  of  Humbert  and  Fernandez

(1977) were obtained from subjects given single 4-hr and 8-hr exposures to 70

and 145 ppm, and 72 and  213 ppm MC, respectively.  Total urinary excretion of

TCE (glucuronide) and TCA  is  observed to be proportional  to the inspired air

concentration of MC and to the expected body burden  from these exposures (with

duration of  exposure also taken into  account).  Both  these research groups

attempted a balance study by estimating the retained body dose of MC either by

measuring lung  clearance during exposure (Monster et al.,  1979b), or by  inte-

grating over the MC alveolar air decay  curve  to infinite  time postexposure

(Humbert and Fernandez,  1977).   The percentages of  the retained dose metabo-

lized to TCE and  TCA were 2.5 and 6.3 percent, respectively.  The difference

can be  ascribed to  the different methodologies used for estimating body dose

and their  inherent imprecisions.   In both studies, the percentages  of MC

metabolized were found  to be  independent of the  retained dose  (Table 4-6).   A

similar observation with rats  was  made by Eben and Kimmerle  (1974),  who
                                i
measured urinary excretion  of TCE and TCA  for 3 days after a 4-hr inhalation

of MC at 221 and 443 ppm.  The amounts of the metabolites excreted were" propor-

tional  to  the  inspired concentration of MC.   In rats,  the urinary ratio of

TCE/TCA was 20/1 rather  than  the 3/1 observed  in humans (Table 4-6).

     A  reasonable conclusion  from these  studies  is that MC  is minimally metabo-

lized by man on the order of  3 to 6 percent of the inhaled  dose.  The percent-

age of  the  dose metabolized  is constant and independent of the body dose (at

least up to  213 ppm),  which suggests  that  some factor  other than the  capacity

of the  metabolizing system is the  limiting one  (e.g.,  hepatic extraction).

4.2.2   Kinetics of  Blood and  Urine Metabolites

     The blood  and  urinary  metabolites of  MC  (total  amount, excretion ratios,

and excretion time-course)  are of interest  as  quantitative  indices of exposure




002MC2/B                             4-21                             11-10-81

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TABLE 4-6.   RELATION BETWEEN INHALATION EXPOSURE AND URINARY METABOLITES OF  MC
Sekl et al.. 1975

Average concentration of metabolites in urine samples from workers daily
exposed to MC.


Plant        Air cone.           No.         TCE*     TCA         TTC/g
	      MC, ppm          Subjects           mg/£           creatinine

  A            0                 30           0        0           0
  B            4.3               10           1.2      0.6         2.1
  C           24.6               26           5.5      2.4         6.8
  D           53.4               10           9.9      3.6        15.0
Monster et al.. 1979b

Averaged amounts of metabolites in total urine collected 70-hr post single
exposure (4 hr).
             Air Cone.           No.
             MC. ppm          Subjects       TCE*     TCA
                                              mg       mg
              72                  6           5.5      1.5
                                *
             145               *  6          11.5      2.8

     Estimated  as % retained dose             2%       0.5%


Humbert and Fernandez, 1977

Averaged amounts of metabolites in total urine collected for 12 days post
single exposure (8 hr).
             Air cond.           No.
             MC, ppm          Subjects      TCE*      TCA
                                             mg        mg
              72                  3          15.2      5.2

              213                  2          30.7     13.0

     Estimated  as % retained dose             4.6*     1.75»
 MCE  found  as  glucuronide.
 002MC2/B                             4-22                             11-10-81

-------
and body  burden.  However,  the  known metabolites of MC--TCE, TCE-glucuronide,

and TCA --  are not  pathognomonic of MC,  but  are  also metabolites of other

chlorinated hydrocarbons, e.g., trichloroethylene.

     From studies of men exposed to 70 and 145 ppm MC inhaled for 4 hr,  Monster

et al.  (1979b)  found that the TCE concentration in blood was proportional  to

both the  inspired concentration  and the blood concentration of MC.  TCE blood

concentration was about  4 percent that of MC (0.2 mg £   and 0.09 mg £    TCE

for 145 and 70 ppm MC inspired, respectively).   After termination of exposure,

blood TCE concentration  declined exponentially  with a half-life of 10 to 12

hr.  Urinary  appearance  of TCE and TCE-glucuronide paralleled the disappear-

ance of blood TCE,  and daily excretion decreased  with a half-time of renal

elimination of  10 to 12 hr.  This value  is  in  agreement with that observed

after ingestion of TCE itself (Briemer et al.,  1974; Muller et al., 1974).   In

contrast to the  first-order blood decay kinetics of TCE, blood concentrations

of TCA progressively increased after  the end of MC exposure for about 40 hr
                               *
before declining exponentially with a  half-life  of 70 to 85 hr.   Consequently,

TCA appeared  in  the  urine in almost equal daily  amounts for 3 days before

decreasing.    Exogenous TCA  administered to men  has a  similarly long half-time

of renal  elimination of  50 to 82 hr,  presumably  because of very tight  non-

covalent binding to  plasma proteins  (Paykoc  and Powell,  1945;  Muller et al.,

1974).   Therefore,  the rise in plasma  concentration of TCA during the 24 to 48

hr period following  exposure is due to a  rate  of  formation of TCA from TCE

greater than  the rate of renal elimination of  TCA.   Similar observations on

daily urinary  excretion  of TCE,  TCE-glucuronide,  and  TCA  following acute MC

inhalation exposure  have been made in  man by  Humbert and Fernandez (1977),  and

in rats by Eben and  Kimmerle (1974).
002MC2/B                             4-23                             11-10-81

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     Stewart et al.  (1969)  investigated  urinary metabolite excretion in men

repeatedly exposed to MC  inhalation  (500 ppm,  7 hr per day for 5 days).   The

daily ratio TCE/TCA during exposure remained relatively constant (about 2.8),

but  rapidly  decreased  within  several  days after exposure  (5th  day,  0.4),

indicating that the daily urinary TCE excretion decreased while the  TCA excre-

tion decreased at  a  lesser  rate or may  have gradually increased.   Eben and

Kimmerle (1974) also chronically  exposed rats  for 14 wk (8 hr daily, 5 days

per week) and  measured  blood levels  of MC, TCE,  and  TCA,  as well as daily

urinary excretion.   Blood concentrations  of MC  and TCE (determined immediately

after daily exposure) remained essentially constant during  the entire 14 week

period, with a concentration ratio of MC:TCE  of 10:1.  Weekly urinary excre-

tion of TCE  reached  a  plateau within 3 to 4 wk, whereas TCA weekly  excretion

was constant throughout the  entire 14-wk period.  The  urinary excretion ratio

TCE/TCA was thus initially low, but after 3 to  4 wk exposure reached a constant

value of approximately 18.
                               «,
     These findings  show that the daily  urinary  ratio of  TCE/TCA (mg/day)

during chronic MC  exposure of  both man and rats is determined by the  relative

rates of metabolic  formation and by  the differences  in  renal elimination of

TCE  (rapid,  half-time 10  to  12 hr) and TCA (slow, half-time 50 to 70  hr).   In

blood, both metabolites achieve steady-state plateau  concentrations  (TCE more

rapidly than TCA)  related to the inspired air  concentration  of MC and  its

metabolites.    In urine, the  TCE/TCA  ratio,  initially  low,  rises  to a  constant

daily value.   After termination of either acute or chronic exposure, the ratio

of the amounts of  daily urinary metabolites,  TCE/TCA (mg/day), is highest 24

hr after exposure.   Thereafter, its progressively decreases daily to  less than

unity by the 5th or 6th day postexposure because of the relatively rapid renal
002MC2/B                             4-24                             11-10-81

-------
elimination of TCE and the slow renal elimination of TCA (Stewart, 1969; Eben



and Kimmerle, 1974; Monster,  1979).



     The profile of TCE and TCA urinary excretion during and after MC  exposure



is similar  to that observed  for  trichloroethylene  (Nomiyama and Nomiyama,



1971; Muller  et  al.,  1974;  Sato et al.,  1977;  Monster  et al.,  1976,  1979a;



Fernandez et  al.,  1977).   However,  the ratio of  TCE/TCA  excreted is  2-fold



greater  for  MC than  for  trichloroethylene, suggesting differences in  the



pathways and  rates  of metabolism to TCE  and TCA for these two chlorinated



hydrocarbons.  Trichloroethylene,  a chlorinated  olefin,  is metabolized by



hepatic  microsomes  to an  epoxide, then to chloral hydrate, and thence to TCE



and TCA  (Liebman and Ortiz, 1977; Henschler, 1977; Van Duuren, 1977).   However,



neither  epoxide nor chloral hydrate have been identified as metabolites of MC.  •



Comparison of the  percentage  of  the  body  dose metabolized  to  TCE  and  TCA  (MC,



3-6 percent  vs.  trichloroethylene,  80-90  percent) explains the  25- to 30-fold



difference in the  total  amounts of these  metabolites (mg/day)  excreted  into



urine  for  comparable  body doses retained  after  inhalation exposure (Seki  et



al.,  1975;  Stewart,  1968;  Stewart et al., 1969; Monster, 1979;  Ikeda and



Ohtsuji, 1972; Ikeda  et al.,  1972).  In a  comparative study of rats exposed to



200  ppm MC or its 1,1,2-isomer,  Ikeda  and Ohtsuji  (1972)  reported that the



urinary  excretion  of  total  chlorinated metabolites  was  substantially  less  for



the  1,1,2-isomer,  indicating  that the 1,1,2-isomer  is metabolized to an even



smaller  extent than MC.   These workers determined the urinary metabolites of



the  1,1,2-isomer  as TCE and  TCA  by  a  spectrophotometric  method based on the



Fujiwara reaction.  However,  the  MC isomer requires  a shift of a chlorine atom



from one carbon to the other  in order  to  form TCA or TCE,  an  unlikely reaction



HI vivo  for  a saturated aliphatic, and  it  is probable that dichlorometabolites




were actually measured.







002MC2/B                            4-25                            11-10-81

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4.2.3  Enzyme Pathways of Methyl  Chloroform Metabolism



     The metabolic pathways and enzyme mechanisms for the aetabolism of halo-



genated hydrocarbons  assume  considerable  importance  for understanding and



assessing cellular toxicity.   Compounds that in the  course of  their  metabolism



form intermediates reactive with  cellular  macromolecules, e.g.,  epoxides or



free radicals, are associated with enhanced cellular toxicity  and carcinogenic



potential  (Van  Duuren,  1977).   In comparison with  other  halogenated  hydro-



carbons, MC  is  not extensively metabolized by  mammalian systems;  this may



explain in part its  lower toxicity and  carcinogenic  potential  (Weisburger,



1977).   Unfortunately,  these same attributes may also have contributed to the



current inadequate state of knowledge  of the pathways of MC biotransformation



and the enzyme systems involved.



     Figure 4-3  summarizes the presently postulated enzyme steps in the bio-



transformation of MC  to TCE  and  TCA,  the only known metabolites  appearing in



the  plasma and  urine of animals  and  man (Table 4-6).   It  is  assumed that

                               c

metabolism occurs principally iti  the liver, although uj vitro  experiments that



directly demonstrate  and evaluate MC  metabolism by the  liver have  only been



reported for TCE production (Ivanetich and Van Den Honert, 1981).



     In view  of  the  absence  of MC-labeled  or  unlabeled  balance  studies with



methodologies of  current  sophistication,  the possibility of the existence of



unknown minor metabolites of secondary pathways  is  real.   Hake et al.  (1960),



in the  only  reported  study using labeled MC, found that about 0.5 percent of


                                                                        14
the dose (of the 2 percent metabolized; see Table 4-7) was converted to   CO-,



following  1,1,1-trichloroethane-l-  C  administration  (intraperitoneal)  to



rats, suggesting  carbon-carbon cleavage to chloroform.  Chloroform is exten-



sively metabolized to COp  in man (50  percent)  and  to glutathione-conjugatpd
002MC2/B                             4-26                             11-10-81

-------
   TCA
       «o
CUC - C
 3     NOH
 "CHLORAL HYDRATE" \^N
7  DEHYDROGENASE
                                 ^
                               Cl - C - CH,
                                 x .    3
                               Cl  I  VV   MICROSOMAL
                                   |  \\ P450 SYSTEM
                                   i    \\ NADPH, r
                                   •      \\
                                   I
                               PEROXISOME
                               OXIDASE,
                               CATALASE
                                SYSTEM
    MICROSOMAL
  DEHYDROGENASE.
       NADP
         CI3C-CH2OH    GLUCURONIYL

      /XMICROSOMAX™™5""*"
      /   ETHANOL   ^
      f   _....__...._    Trc.ftinriior
 ETHANOL   ^
OXYGENASE    TCE•6LUCURONIDE
 NADPH O-
                                        NADPH, 0
                                         'ALCOHOL
 trj ALCOHOL
 * DEHYDROGENASE?
O
                     ci3c - c

                      CHLORAL
    Figure 4-3. Postulated pathways of hepatic biotransformation of MC.

    Source: Davidson (1980) and Ivanetich and Van Den Honen (1981).
                            4-27

-------
chloromethyl derivatives (Fry et  al.,  1972).   During these  conversions,  free



radicals or reactive intermediates are  produced which contribute  to  chloroform


                                                            14
toxicity (Brown et  al.,  1974).   However,  the origin of the   CO- is an open



question since Hake et al. did not identify chloroform as a metabolite of MC;


          14
also, the   C-MC  used  was  contaminated with trace amounts  of  the MC isomer



l,l,2-trichloroethane-l-14C (0.4  percent)  and  l,l-dichloroethane-l-14C (0.5



percent).  Both of  these compounds were reported by  Van Dyke (1971, 1977) to



be readily dechlorinated by the rat liver microsomal  P450 system.  The optimal



configuration for dechlorination  was a dichloromethyl group.  Hence, 1,1,2-



trichloroethane, the more  toxic  isomer, was readily  dechlorinated but 1,1,1-



trichloroethane was not.   For  the isomer 1,1,2-trichloroethane,  the products



were  identified as  mono-and dichloroethanol and mono- and dichloroacetic  acid



(Van  Dyke and  Wineman,  1971),  which are known to be  metabolized to  CO- or to



form  glutathione conjugates (Yllner, 1971a,b).   Carlson (1973) found that pre-



treatment of rats with phenobarbital,  but  not  methylcholanthrene, potentiated
                               ^


hepatotoxicity (measured as  serum SCOT and SGPT)  of both inhaled MC and  its



1,1,2-isomer, but the  toxicity of the  isomer  was  increased to a  far greater



extent.  These  several  observations suggest that  1,1,2-trichloroethane,  but



not  MC,  may be  extensively metabolized by  the  oxidative dechlorination  system



of Van Dyke and Wineman  (1971).



      Figure 4-3  shows  that the initial pathway  of MC transformation is  hy-



droxylation of  MC to TCE by  the  microsomal P450 mixed function oxidase  system



(Ivanetich  and  Van  Den  Honert, 1981).   There  are  several other  observations



which support this  finding.



      First, when  MC is  incubated  aerobically  with rat  liver microsomes plus



NADPH,  it produces  a P450  type I  binding spectrum  with  peaks at 452 nm  and  420



nm,  and  a reduced spectrum with a peak at  420  nm only (Cox  et  al. 1976;  Pelkonen



and  Vainio, 1975).



002MC2/B                            4-28                              11-10-81

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     Secondly, MC inhalation induces the P450 drug metabolizing system (Fuller



et al.,  1970;  Lai  and Shah, 1970).  Rats exposed to inhaled MC (2500 to 3000



ppm) for  24  hr exhibited decreased hexobarbital sleeping time, and decreased



duration of effect of meprobamate and zoxazolamine.   Furthermore, liver micro-



somes  from  MC-pretreated rats  exhibited increased  u>  vHro metabolism of



hexobarbital,  zoxazolamine,  and aminopyrine,  type I substrates.  Livers from



pretreated rats had increased cytochrome P450 and NADPH cytochrome C reductase



activity, preventable by cycloheximide  or actinomycin D.   This  induction  of



hepatic microsomal drug  metabolism in the reports of Fuller et  al.  (1970)  and



Lai  and  Shah  (1970),  suggests the possibility  that  MC  might induce its own



metabolism with repeated daily  exposure.  Stewart et al.  (1969)  observed that



men  experimentally exposed  to 500 ppm MC 7  hr  for 5 days excreted in their



urine  progressively  increased daily amounts  of  TCE and TCA  although the daily



ratio  TCE/TCA  remained  relatively constant (2.8). Eben  and Kimmerle (1974)



found  that  rats exposed  daily to 204 ppm MC (8 hr/day 5 day/wk) for 14 weeks



excreted progressively increasing amounts of TCE for the first few weeks.   The



TCE/TCA ratio  increased  to  a maximum (about 20) that was maintained for the



remainder of the 14-wk exposure period.



     Thirdly,  MC can  interact  with pathways of drug metabolism. Plaa et al.



(1958) and  Shah and  Lai  (1976) found that MC given intraperitoneally to mice



(1 mg/kg)  potentiates pentobarbital sleeping time  and  reduces hexobarbital



metabolism by  liver microsomes.   This observation, in contrast to induction of



metabolizing enzymes  by  inhalation exposure or  by intraperitoneal administra-



tion in oil, is explained on the basis of different amounts of MC reaching the



liver  by  the  various routes and methods of administration.   Van  Dyke  and



Rikans (1970)  added  MC  directly to rat  liver microsomes  incubated  in vitro.
002MC2/B                             4-29                             11-10-81

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They  observed  no  effect on N-demethylation of aminopyn'ne, but a 50 percent

increase of  aniline  4-hydroxylation,  a type II substrate.  The 1,1,2-isomer

produced similar results.

     The origin of TCA  as  a plasma and urinary metabolite of MC is postulated

to  occur  from enzymic  oxidation of TCE.   TCE-glucuronide,  also present in

plasma and urine,  is presumed to be  formed by glucuronyl transferase.  TCE,

exogenously  administered to man, yields TCA as  a  metabolite (Marshall and

Owens, 1954; Owens and  Marshall, 1955a,b; Muller et al., 1974).  Marshall and

Owens (1954) also observed TCA formation during TCE incubation with rat or dog

liver slices.  The most likely reaction for the oxidation of TCE to TCA would

involve the  enzyme  alcohol dehydrogenase  (Figure  4-3).   However,  ijri vitro

studies with alcohol  dehydrogenase  purified from horse liver,  and  with rat

liver cytosol  fractions,  have shown that TCE is a  poor substrate for alcohol

dehydrogenase and that significant conversion to trichloroacetaldehyde (chloral)

does  not occur (Sellers et al.,  1972; Friedman and Cooper, 1960; Marshall and
                                *
Owens, 1954).  The reverse reaction —reduction of chloral to TCE — proceeds

rapidly with a Km of 2.7 x 10~3M for horse liver enzyme (Butler, 1948, 1949;

Marshall  and Owens,  1954,  1955a,b; Friedman and Cooper, 1960; Sellers et al.,

1972).  Also,  chloral hydrate has been sought, but not detected, as an  inter-

mediate metabolite  in  plasma of  rats and man exposed by  inhalation  to MC

(Monster et  al.,  1979;  Eben and Kmmerle, 1974).  On the other  hand, chloral

hydrate exogenously  administered  is  very rapidly metabolized jjn vivo with  a

half-life of only  a  few minutes, yielding both TCE and TCA as metabolites in

plasma and urine  (Butler,  1948;  Marshall and Owens, 1954,  1955a,b;  Breimer  et

al., 1974;  Muller et al., 1974;  Cole et al., 1975).
002MC2/B                             4-30                             11-10-81

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     The conversion of chloral to TCA is usually ascribed to so-called "chloral
hydrate dehydrogenase," a substrate-specific NAD-dependent enzyme described by
Cooper and Friedman (1958).   This enzyme was obtained from rabbit liver acetone
powders.  Cooper and  Friedman reported  that acetaldehyde was not a substrate,
but rather a  markedly effective inhibitor; they did not demonstrate  a normal
endogenous substrate.   Human cytosolic acetaldehyde  dehydrogenase does not
convert chloral hydrate  to TCA  (Kraemer and Deitrick, 1968; Blair and Bodley,
1969; Sellers  et al.,  1972).  Grunnet (1973) reported that chloral hydrate  is
not a substrate for mitochondrial  NAD-dependent acetaldehyde dehydrogenase.
Microsomal NADP-dependent  acetaldehyde  dehydrogenases  with  broad substrate
specificity have been described,  but whether chloral hydrate is a substrate
has not been determined (Tottmar et al., 1973).
     In short, there  is  little  evidence which precisely defines the enzymatic
pathways for the metabolism of MC to TCE by P450 mixed function oxidase system,
and TCE to TCA by classical alcohol dehydrogenase and acetaldehyde dehydrogenase
enzymes.  Although  as yet  unintestigated,  it is possible  that  TCE may be a
substrate for the microsomal  P450 ethanol  oxidative system (MEOS) described by
Lieber  and his coworkers for  ethanol and other  alcohols  (Lieber and De  Carli,
1969; Teschke  et al., 1977).   TCE has been shown by Uehleke et al.  (1976) to
give a  binding spectrum with rat  microsomes.   An  alternative pathway  also
uninvestigated is  afforded by the  peroxisomal oxidase-catalase  system  (Figure
4-3), which is known  to  readily oxidize ethanol  and a broad spectrum of other
substrates (Masters and  Holmes, 1979;  Chance et al., 1977).   Indeed,  peroxi-
somes may perhaps  be  involved in the initial oxidation  of  MC to TCE and other
halogenated hydrocarbons as well.
002MC2/B                             4-31                             11-10-81

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 4.3  SUMMARY AND  CONCLUSIONS


      The pharmacokinetics and metabolism of MC in man have been studied less


 than other  chlorinated  aliphatic  solvents,  both  in controlled experimental


 conditions  and in the workplace.   Like other solvents of this group, inhala-


 tion and lung absorption of  MC  vapor in the air is  the  most important and


 rapid route  of  absorption  into the  body.   Absorption  through the skin by


 direct liquid contact  is slow and  adds less  than 5 percent to  total body dose.


 At the accepted TWA  value (350 ppm) for an 8-hr exposure, less than 2 g may be


 expected to be absorbed  into the  body of a  normal 70 kg man.  This is because


 pulmonary absorption is  directly  related to the  blood/air  partition coeffi-


 cient, which  for MC is  less  than that  of  most other structurally related


 solvents.   MC total  body dose increases  in  direct proportion  to inspired  air


 concentration and duration  of exposure; it is also increased by physical


 activity during exposure.  MC distributes throughout the body, readily cross-


 ing the blood-brain barrier  and probably  the placenta! barrier as well.   It
                                *

 can be assumed that MC also  distributes into the colostrum or milk of nursing


 mothers, although no specific data are  available.   Relative  body  tissue con-


 centrations are  not  known,  but there is  a high  affinity for adipose  tissue due


 to the higher lipid/  blood  partition coefficient  of MC compared with that of


 other related  solvents.   Blood  and  tissue  concentrations achieved  during


 exposure are  directly  proportional to inspired air concentration and  total


 body dose.


      MC is metabolized in man to a very limited extent—about 6 percent of the


'total body dose.   Metabolism appears to occur principally in  the  liver, and to


 an unknown  extent in  other  tissues, yielding only trichloroethanol  (ICE.)  and


 trichloroacetic acid (TCA)  as identified  metabolites.   Urinary excretion  of


 these metabolites is proportional  to inspired air concentration and  the total





 002MC2/B                             4-32                              11-10-81

-------
body dose of  MC.   Detailed information concerning the enzymatic pathways of



metabolism is lacking.   The importance of greater knowledge of  the  biochemical



mechanisms and  pathways of metabolism  relates  to awareness that "reactive



intermediate" metabolites  may  contribute to tissue and organ  toxicities  as



well as  carcinogenic potential.  The mechanism(s) of the initial biotransfor-



mation of MC  to TCE,  where "reactive  intermediates" may occur, are specula-



tive, although  present  evidence  suggests a microsomal  cytochrome P450 oxida-



tive reaction.  However, specific covalent binding studies  which would confirm



reactive  intermediates  have not  been carried out.  Also, other minor pathways



and unidentified metabolites cannot be excluded because of  the lack of defini-



tive studies.  Metabolism  is enhanced by microsomal inducers such as phenobar-



bital and possibly MC  itself.   The interaction of MC metabolism and toxicity



with other common  drugs,  including ethanol, has not been adequately investi-



gated.



     During post-exposure, more than 80 percent of MC is excreted unchanged by



the  lungs.   Alveolar  air  concentration and blood  concentration  decline  in  a



parallel  exponential fashion exhibiting three  major  components of  elimination



with half-times of approximately  1,  9, and 30  hr.  The  long half-time of



elimination  (30 hr) is  associated with  elimination  from  adipose tissue and



indicates that  accumulation may  occur  with repeated  daily  exposures,  particu-



larly in obese  persons.  Further research  on  the disposition  and  fate of  MC



after low chronic  vapor exposures  would be helpful,  particularly with respect



to  bioaccumulation, enzyme mechanisms of  biotransformation, and interactions



with common drugs.
 002MC2/B                             4-33                              11-10-81

-------
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002MC4/C                             4-34                        11-10-81

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•m
002MC4/C                             4-35                         11-10-81

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     mammalian tissues.   Trends.  Biochem.  Sci. 4:233-236,  1979.

Miller, K. W. , W. D. M.  Paton, E.  B.  Smith, and R.  A. Smith.  Physicochemical
     approaches to  the mode  of action of general  anesthetics.   Anesthesiology
     36:339-351, 1972.
002MC4/C                             4-37                         11-10-81

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Monster, A. C.   Difference  in uptake, elimination and metabolism in exposure
     to  trichloroethylene,  1,1,1-trichloroethane  and tetrachloroethylene.
     Int. Arch. Occup. Environ.  Hlth. 42:311-317, 1979.

Monster, A.  C.,  and  G.  Boersma.   Simultaneous determination of trichloro-
     ethylene  and metabolites in blood and exhaled air by gas chromatography.
     Int. Arch. Occup. Environ.  Hlth. 35:155-163, 1975.

Monster, A. C. ,  and J. M. Houtkooper.   Estimation  of individual uptake  of
     trichloroethylene,  1,1,1-trichloroethane and  tetrachloroethylene from
     biological  parameters.   Int.  Arch.  Occup.  Environ. Hlth.  42.319-323,
     1979.

Monster, A. C. ,  G.  Boersma,  and W.  C. Duba.   Pharmacokinetics  of trichloro-
     ethylene  in volunteers; influence of workload and exposure concentration.
     Int. Arch. Occup. Environ.  Hlth. 38:87-102, 1976.

Monster, A. C.,  G.  Boersma, and W. C. Duba.   Kinetics of trichloroethylene  in
     repeated  exposure of volunteers.  Int. Arch. Occup. Environ. Hlth. 42:283-
     292, 1979a.

Monster, A. C., G. Boersma, and H.  Steenweg.   Kinetics of 1,1,1-trichloroethane
     in  volunteers;  influence of  exposure concentration and work load.   Int.
     Arch.  Occup. Environ. Hlth.  42:293-301,  1979b.

Morgan,  A., A. Black, and D.  R.  Belcher.  Studies on the absorption of halo-
     genated  hydrocarbons and their excretion in breath using  3&C1  tracer
     techniques.   Ann. Occup.  Hyg.  15:273-283, 1972.

Miiller,  G., M. Spassovski,  and p.  Henschler.   Metabolism of trichloroethylene
     in  man.   II.  Pharmacokinetics  of metabolites.   Arch.  Toxicol. 32:283-295,
     1974.

Nomiyama,  K. , and  H.  Nomiyama.   Metabolism  of  trichloroethylene  in  human.
     Int. Arch.  Arbeitsmed. 28:37-48, 1971.

Ogata,  M. ,  K. Tomokuni,  and  H. Asahara.   Simple microdetermination of tri-
     chloroethanol  glucuronide and trichloroacetic acid in urine.   Int.  Arch.
     Arbeitsmed. 32:203-215,  1974.

Owens,  A.  H., Jr.,  and E. K.  Marshall,  Jr.   A comparison of the metabolism of
     ethanol  and trichloroethanol.   Bull.  Johns Hopkins  Hosp.  9_7:395-404,
     1955a.

Owens,  A.  H. , Jr.,  and  E. K. Marshall,  Jr.   Further  studies  on the metabolic
     fate  of  chloral  hydrate  and  trichloroethanol.   Bull. Johns Hopkins Hosp.
     97:320-326, 1955b.

Paykoc,  2.  and J.  F.  Powell.  The excretion  of  sodium  trichloracetatc.   J.
     Pharmacol.  Expt.  Ther. 85:289-293,  1945.

Pelkonen,  0., and H.  Vainio.  Spectral  interactions  of  a series of  chlorinated
     hydrocarbons with cytochrome P-450  of  liver microsomes from  variously-
     treated  rats.   FEBS Letters 51:11-14,  1975.
 002MC4/C                             4-38                        11-10-81

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Plaa, G. L. ,  E.  A.  Evans, and C. H. Hine.  Relative hepatotoxicity of seven
     halogenated hydrocarbons.  0.  Pharmacol.  Exp.  Ther.  123:224-229, 1956.

Prendergast, J. A.,  R.  A.  Jones, L. J.  Jenkins, Jr., and J.  Siegel.   Effects
     on experimental  animals of  long-term  inhalation  of trichloroethylene,
     carbon tetrachloride, 1,1,1-trichloroethane, dichlorodifluoromethane, and
     1,1-dichloroethylene.  Toxicol. Appl. Pharmacol. 10:270-289, 1967.

Rowe, V. K., T. Wujkowski, M. A.  Wolf,  S.  E. Sadek, and R. D. Stewart.  Toxicity
     of a solvent mixture of 1,1,1-trichloroethane and tetra chloroethylene as
     determined by experiments on laboratory animals and  human  subjects.   Am.
     Ind.  Hyg. Assoc. J. 24:541-554, 1963.

Salvini, M.   Psychological  Effects  of  Trichloroethylene and 1,1,1-Trichloro-
     ethane Upon Man.  Behavioral Toxicology, U.S. Dept. HEW, Publ. No.  (N10SH)
     74-126, 1974.

Salvini, M. ,  S. Binaschi,  and M.  Riva.  Evaluation  of  the psychophysiological
     functions  in  humans exposed to the  "threshold limit value"  of  1,1,1-
     trichloroethane.  Brit. J.  Ind. Med.  28:286-292, 1971.

Sato, A. ,  and T. Nakajima.   A structure-activity relationship of some chlori-
     nated hydrocarbons.  Arch.  Environ. Hlth.  34:69-75, 1979.

Sato, A.,  T.  Nakajima,  Y.  Fugiwara, and N.  Murayama.   A pharmacokinetic model
     to study the  excretion  of  trichloroethylene and  its metabolits after
     inhalation exposure.  Brit.   J.  Ind. Med. 34:56-63, 1977.

Savolainen, H. , Pfaffli,  P.,  Tengen, M. and Vainio,  H.   Trichloroethylene and
     1,1,1-trichloroethane:  effects on brain and liver after five days inter-
     mittent  inhalation.  Arch.*Toxicol. 38:229-237, 1977.

Schwetz, B. A.,  B.  K. J. Leong,  and P. J. Gehring.  The effect  of maternally
     inhaled  trichloroethylene,   perchloroethylene,  methyl   chloroform, and
     methylene chloride  on embryonal  and fetal  development  in mice and rats.
     Toxicol.  Appl. Pharmacol. 32:84-96,  1975.

Seki, Y. ,  Y.  Urashima,  H.  Aikawa, H. Matsumura,  Y.  Ichikawa, F.  Hiratsuka, Y.
     Yoshioka, S.  Shimbo,  and M. Ikeda.  Trichloro-compounds in the  urine of
     humans exposed  to  methyl chloroform  at sub-threshold levels.  Int. Arch.
     Arbeitsmed. 34:39-49, 1975.

Sellers, E. M. ,  M.  Lang, J. Koch-Weser,  E.  LeBlanc, and H.  Kalant.   Interac-
     tion  of  chloral hydrate  and  ethanol  in  man.  I.   Metabolism.  Clin.
     Pharmacol. Therap.  13:37-49, 1972.

Shah, H. C. ,  and H.   Lai.   Effects  of  1,1,1-trichloroethane administered  by
     different routes and in different solvents on barbiturate hypnosis  and
     metabolism in mice.  J. Toxicol. Environ.  Hlth. 1:807-816,  1976.

Stahl,  C.  J. , A.  V.   Fatteh, and  A. M.  Dominguez.   Trichloroethane poisoning:
     observations  on the pathology and toxicology in  six fatal cases.   J.
     Foren. Sci. 14:393-397, 1969.
002MC4/C                             4-39                         11-10-81

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Stewart,  R.  D.   Methyl  chloroform  intoxication:   diagnosis and treatment.
     JAMA 215:1789-1792, 1971.

Stewart,  R.  D.   The  toxicology of 1,1,1-trichloroethane.  Amer. Occup. Hyg.
     11:71-79, 1968.

Stewart, R. D., and J.  T. Andrews.  Acute intoxication with nethyl  chloroform.
     JAMA 195:904-906,  1966.

Stewart, R. D. and H. C. Dodd.   Absorption of carbon tetrachloride, trichloro-
     ethane,  tetrachloroethylene,  methylene chloride,  and 1,1,1-trichloro-
     ethane through the human skin.   Am. Ind. Hyg.  Assoc. J. 25:439-446, 1964.

Stewart, R. D., H. H. Gay, D. S. Erley, C. L. Hake, and A. W.  Schaffer.  Human
     exposure to 1,1,1-trichloroethane vapor:  relationship of expired air and
     blood concentrations  to exposure and toxicity.  Am. Ind. Hyg. Assoc. J.
     22:252-262, 1961.

Stewart,  R.  D. , H.  H.   Gay,  A.  W. Schaffer, D. S.  Erley, and V.  K.  Rowe.
     Experimental  human  exposure  to methyl chloroform vapor.   Arch. Environ.
     Hlth. 19:467-472,  1969.

Tanaka, S.,  and M. Ikeda.   A method for determination  of trichloroethanol  and
     trichloroacetic acid in urine.   Brit.  J.  Ind. Med.  25:214-219,  1968.

Teschke,  R.,  K.  Ohnishi, Y. Hasumura,  and  C.  S. Lieber.   Hepatic  microsomal
     ethanol oxidizing system:   Isolation and reconstitution.   In:   Microsomes
     and  Drug  Oxidations., Proc.  3rd Int.  Symp. Berlin,  1976,  Suppl.  Biochem.
     Pharmacol. Pergammon, Oxford, pp.  103-110, 1977.
                                *
Torkelson, T.  R. ,  F. Oyen,  D.  «0. McCollister,  and V.  K.  Rowe.  Toxicity of
     1,1,1-trichloroethane  as  determined  on laboratory  animals  and  human
     subjects.  Am.  Ind. Hyg. Assoc. J. 19:353-362, 1958.

Tottmar,  S.  0., H.  Pettersson, and K.  H. Kiessling.  The  subcellular dis-
     tribution and properties of  aldehyde dehydrogenases  in rat  liver.  Biochem.
     J. 135:577-586, 1973.

Uehleke,  H. , S. Tabarelli-Poplawski, G.  Bonse, and D. Henschler.    Spectral
     evidence  for 2,2,3-trichlorooxirane  formation during microsomal  tri-
     chloroethylene  oxidation.   Arch. Toxicol. 37:95-105,  1977.

Vainio,  H. ,  H.  Savolainen and P.  Pfaffli.   Biochemical  and  toxicological
     effects  of combined  exposure  to  1,1,1-trichloroethane  and trichloro-
     ethylene on  rat liver and brain.   Xenobiotica  8:191-196,  1978.

Van  Duuren,  B.  L.   Chemical structure,  reactivity and  carcinogenicity  of
     halohydrocarbons.    Environ.  Hlth.  Persp.  21:17-23,  1977.

Van  Dyke,  R. A.  Dechlorination  mechanisms  of chlorinated olefins.   Environ.
     Hlth. Persp.  21:121-124, 1977.

Van  Dyke, R. A., and  L.  E.  Rikans.  Effect of the volatile  anesthetics on
     aniline hydroxylase  and  aminopyrine demethylase.   Biochem.  Pharmacol.
     19:1501-1502, 1970.


002MC4/C                              4-40                        11-10-81

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Van Dyke, R. A., and C. G. Wineman.  Enzymatic dechlorination:  dechlorination
     of chloroethanes  and propanes £n  vitro.   Biochem.  Pharmacol.  20:463-470,
     1971.

Weisburger,  E.  K.   Careinogenicity studies  on halogenated  hydrocarbons.
     Environ. Hlth. Persp. 21:7-16, 1977.
                                                 14
Yllner,  S.   Metabolism  of  1,2-dichloromethane-  C in  the  House.  Acta.
     Pharmacol. et Toxicol. 30:257-265, 1971a.

    ?r, S.   Metabolism of chloro<
     et Toxicol. 30:69-80, 1971b.
                                           14
Yllner, S.  Metabolism  of chloroacetate-1-  C in the mouse.  Acta Pharmacol.
002MC4/C                              4-41                        11-10-81

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                               5.  TOXIC EFFECTS
     Since  its  commercial  introduction in 1954, methyl  chloroform  has been

used increasingly  as  an industrial solvent and in consumer products.  Usually

the most  important path of entry into the body is by inhalation.   The health

hazards of  MC  inhalation in the work place or public environment, as  well as

exposure by other  routes, are pointed out by human and animal studies  indicat-

ing that  MC has  significant adverse biological effects on the central  nervous

(CNS)  and cardiovascular systems.  In addition, MC  may  cause organ tissue

damage  generally associated with  chlorinated  hydrocarbon  solvent toxicity.

Like some of  these other structurally related compounds, it may also  have a
                               %
teratogenic, mutagenic,  and/or'carcinogenic potential.   The available  litera-

ture on the possible toxic effects of MC is reviewed in this chapter.

5.1  HEALTH EFFECTS IN HUMANS

5.1.1  Experimental Studies

     Experimental  studies  in humans have centered around three general areas:

clinical experiences with  MC  as  an anesthetic; the kinetics of MC absorption

and excretion after exposure via  the  inhalation and cutaneous routes;  and the

impairment of  psychophysiological functions in humans exposed to MC.

     Dornette  and  Jones (1960)  administered  MC to  50 patients  undergoing

elective surgery.  Nitrous oxide-oxygen (4:1)  was used as the vehicle  and as  a

supplemental anesthetic agent.   The concentration of MC required for induction
002MC3/A                             5-1                              11-10-81

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of surgical plane anesthesia varied from 10,000 to 26,000 ppm,  and for  mainten-
ance of  light  anesthesia  from 6,000 to 22,500 ppm.  Rapid induction and re-
covery, analgesia, and  the  absence of disagreeable odor, respiratory depres-
sion, postoperative depression, nausea and vomiting were listed as advantages
in the use  of  MC.   A definite disadvantage was depression of blood pressure
during anesthesia  of moderate depth.  The  tendency to develop ventricular
arrhythmias during hypoxia  was  also observed.  (This effect was reversed when
effective oxygenation was reestablished).
     Cardiac sensitization  by halogenated hydrocarbons, with  resultant in-
creased  susceptibility  of the heart  to  catecholamine-produced arrhythmias,
e.g.,  ventricular  fibrillation or ventricular tachycardia,  is  a  well-known
phenomenon recently reviewed by Aviado et al (1976).  Clark and Tinston (1973)
believe  that cardiac sensitization caused by  these  relatively inert,  lipid
soluble  hydrocarbons  is very likely to be a structurally non-specific  action
on myocardial  membranes by  solution and distribution within these membranes,
and  is,  therefore, an example or  "physical toxicity" (see  Sect. 5.2.3).  From
an evaluation of 14 halogenated hydrocarbons in dogs, Clark and Tinston (1973)
found  that  the cardiac  sensitizing potencies, as  determined by the partial
pressure  in inhaled  air needed to  sensitize  the  heart to epinephrine,  was
directly  related  to  their saturated vapor pressures.   In  a  similar way, the
anesthetic  potency  or  narcotic  action of  the halogenated hydrocarbons, a
structurally nonspecific  action,  is  directly related  to  the  blood/air or
lipid/air partition coefficient for these compounds (Table 4-1) (Miller et  al.,
1972; Eger et al., 1965; Sato and Nakajima, 1979).
     Siebecker  et  al.  (1960) reported that electroencephalographic patterns
during MC anesthesia showed little change before  circulatory  depression and
that  the changes were  similar to  those  during  halothane anesthesia.   The

002MC3/A                             5-2                               11-10-81

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investigators found MC to be  less potent clinically than either chloroform or
halothane in supplementing nitrous oxide-oxygen for anesthesia.
     Stewart et al.  (1961)  evaluated the acute effects of increasing concen-
trations (0 to 2,600 ppm) of MC over a 15 min exposure period.   The commercial
grade of MC  used was chlorothane.  Table 5-1 shows the subjective and physio-
logical responses during  exposure  to MC.   In another  experiment  in which 6
subjects were exposed  to  MC at 500 ppm for 78 min or 186 min,  no eye irrita-
tions  or  dizziness  occurred,  nor  were balance  or coordination affected.
Exposure at 900 to 955 ppm for 73, 35, and 20 min produced a number of psycho-
physiological effects which are listed in Table 5-2.
     In another study, Torkelson  et al.  (1958) exposed humans  to MC (Chloro-
thane) vapors.  Exposure  to 550 ppm for 90 minutes had no measureable effect
on  the  vital signs being monitored.   Exposure to 500 ppm  for  450 minutes
produced no significant changes in pulse, respiration, blood pressure, reflexes,
or equilibrium;  liver function tests were also negative.   Exposure to 1000 ppm
for 30 minutes was also without*effect.  However,  exposure  to 900 to  1000 ppm
for 75 min resulted in slight eye irritation and a feeling of lightheadedness,
and  the Flanagan and  Romberg tests revealed a  slight but definite loss of
coordination and  equilibrium.   ECG  and  liver, function  tests  were normal.
Exposure to  1900  ppm  for 5 minutes resulted in  an  obvious disturbance of
equilibrium and a positive Romberg test.
     In 1969, Stewart  et  al.  reported  that exposure to 500  ppm  for  periods of
6.5 to 7 hours/day  for 5 consecutive  days  resulted  in mild subjective  re-
sponses (sleepiness, eye  irritation,  and mild headache).   The  only untoward
physiological response was  an abnormal Romberg  test.  None of  the  clinical
tests performed during or following the exposure were abnormal.
002MC3/A                             5-3                              11-10-81

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             TABLE 5-1.   SUBJECTIVE AND PHYSIOLOGICAL RESPONSES TO A
                 CONSTANTLY INCREASING METHYL CHLOROFORM VAPOR
                    CONCENTRATION OVER A PERIOD OF 15 MINUTES
Concentration
    (ppm)                          Responses to Exposure


   0 to 1000             Increasing awareness of a slightly sweet, not unpleasant
                          odor.

1000 to 1100             Mild eye irritation noted in 6 of 7 subjects.

1900 to 2000             6 of 7 subjects aware of throat irritation.

    2600                 1 subject very lightheaded.

    2650                 2 subjects unable to stand.   3 subjects very lightheaded,
                          but able to stand.  2 subjects were not lightheaded, and
                          one of these was able to demonstrate a normal Romberg
                          test.
Source:  Stewart et al. (1961).
 002MC3/A                             5-4                              11-10-61

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              TABLE 5-2.   SUBJECTIVE AND PHYSIOLOGICAL RESPONSES TO
                    METHYL CHLOROFORM VAPOR CONCENTRATIONS
                               OF 900 TO 1000 PPM
  Average
Concentration
               Responses to Exposure*
900 ppm for 20
    minutes
(3 subjects)
910 ppm for 35
    minutes
(2 subjects)
951 ppm for 73
    minutes
(3 subjects)
Positive Romberg in one subject.   Greater effort re-
 quired to perform a normal Romberg in two subjects
 after 10 minutes of exposure.   Heel-to-toe walking
 normal.  Two subjects experienced lightheadedness
 above 900 ppm.

Greater mental efforts required to perform a normal
 Romberg test after 10 minutes of exposure.  All
 heel-to-toe walking performed well.   One subject
 experienced persistent lightheadedness above
 900 ppm.

Greater mental effort required to perform a normal
 Romberg test after 10 minutes of exposure.  After
 15 minutes of exposure one subject had consistently
 positive Romberg.  Heel-to-toe walking performed
 well by all during exposure.   No lightheadedness.
aMild eye irritation was noted by all subjects when vapor concentrations rose
 above 1000 ppm.

Source:   Stewart et al. (1961).
002MC3/A
            5-5
11-10-81

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     In 1975, Stewart et al. reported an experiment in which 20 subjects were
exposed to 500  ppm  MC  or less for 7.5  hours  per day,  5 days per  week for 3
weeks.   No serious  deleterious  effects  upon health or performance were  de-
tected, and  the health  of the subjects remained  unimpaired  during the in-
halation studies.   The  blood  chemistries,  hematologies, urinalyses,  electro-
cardiograms,  and pulmonary function tests  remained normal.   There  was a slight
increase in  the number  of reported negative subjective responses  (Table 5-3).
     Twelve  subjects were exposed to 250,  350,  450,  and 550 ppm  of  MC  in
inspired air  during four  continuous  30 minute periods  in  an experiment  re-
ported by Gamborale and Hultengren (1973).  The air-gas mixture was  supplied
via a  breathing valve and a mouthpiece with very  low resistance.  The  effects
of the  introduction of  the breathing tube  were not assessed.  In  the  final  20
minutes of each exposure period, five performance tests were made.  Two of the
tests were of perceptual  speed and the others were tests  of simple  reaction
time, choice  reaction time, and manual  dexterity.  The same subjects were also
studied under control conditions  in which  inspired air  contained  no  MC but  in
which  all operations and  measurements were the same as  during exposure to the
solvent.  The presence  or absence of MC was completely disguised by the use  of
menthol crystals.   To  balance the training effects between  experimental  and
control conditions,  the order of conditions was reversed for  half the  subjects.
     The change in  mean performance level  during exposure to the increasing
concentrations  of MC differed systematically from the  change in  performance
under  control  conditions.   The level  of performance  in the  manual dexterity
test and two  perceptual tests were affected by training; however,  the  training
effect  was  less pronounced  during exposure to MC.  The tests of reaction time
were less sensitive to  training and, with  these,  there was an absolute decline
in performance  capability as the  exposure concentration increased.   However,

002MC3/A                             5-6                               11-10-81

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TABLE 5-3.   NUMBER OF SUBJECTIVE RESPONSES TO METHYL CHLOROFORM EXPOSURE



in
i





i— •
i
o
i
00


Lightheadedness
Syncope
Mild sleepiness
Mild eye irritation
Mild nose or throat
irritation
Mild headache
Nausea
Fatigue
Objectionable odor

Source: Stewart et




0
0
0
0
0
0
0
0
1
0

al.





10 Males
ppm 100 ppm 350 ppm 350 ppm 500 ppm
0
0
0
0
0
0
0
0
0

001000011000000000000
000000000000000000000
011100130002000001000
oooooooo o''o oiooooooooo
011000000001200000000
001001011001000000000
000000000000000000000
000000000000000000000
000000000000000000000


10 Females
0 ppm 350 ppm
00 10000
00 00000
00 00000
00 00000
10 10000
20 01110
00 00000
00 00000
00 99999

(1975).







-------
this study contained  several  drawbacks:  e.g.,  the substance used to disguise
the MC odor  may  itself have had a toxic  effect,  and the introduction of a
breathing tube may have induced stress in the subjects.   Nevertheless, statis-
tically significant performance differences  between  experimental and control
conditions were obtained  for  all  tests with exposures  at  350 ppm or more.
     Salvini  et  al.  (1971) evaluated psychophysiological effects  after  ex-
posing six male  university students  to an average vapor concentration of 450
ppm MC for two  periods of 4 hours,  separated  by  a 1.5 hour interval.  Each
subject was  examined  on  two different days  using a crossed-scheme analysis.
The psychophysiological tests  included  a perception test with tachistoscopic
presentation, the Wechsler Memory  Scale test,  a complex reaction time test,
and a manual  dexterity test.   After two  exposures, no  disturbances  in motor
function, coordination, equilibrium,  or behavior patterns were observed in any
of  the subjects.   However, there  were some  complaints  about  eye  irritation.
The only  factor  that was  reported to  be statistically significant was  the
association  between exposure  to MC and perception of  mental  strain.   Under
stress conditions, exposure to 450 ppm MC decreased perceptive capabilities.
A  small  reduction  in  performance  was also observed but was not statistically
significant.
     Savolainen et al.  (1981)  evaluated  various psychophysiological  functions
in  9 male students exposed for 4 hours/day,  at 6-day intervals, to MC (200 and
400 ppm)  and m-xylene (200 ppm) and  MC  (400  ppm)  in  combination.   Exposure  to
MC  or  MC and  xylene  together were  without  effect on  reaction  time,  body
balance, and critical flicker  fusion thresholds.
5.1.2  Occupational Studies
     Chronic  occupational exposure  to  other chlorinated solvents  has been
associated with  adverse neurological  and behavioral  effects.   However,  it has

002MC3/A                             5-8                              11-10-81

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not been  specifically  determined  whether relatively  low level  exposures  to MC
are clearly associated with such effects.
     A  recent attempt to assess  the  central  and peripheral nervous  system
effects of  MC in occupational  situations  was  undertaken by Maroni and  co-
workers (1977).   They studied  a  very small group  (22  subjects) of female
workers exposed  to  MC  vapors at concentrations  ranging from 110 to 990  ppm.
When compared to workers who were reportedly  unexposed,  no differences  were
found in clinical symptoms or measures of nerve conduction velocity and psycho-
metric function.
     In another  study,  Seki  et al. (1975) surveyed 196 male workers employed
in four Japanese printing factories  where MC was the sole organic solvent  in
use.   The four groups  of workers were exposed  to average concentrations of
4, 25, 28, and 53 ppm.   The workers participated  in a medical interview coupled
with a test for  sense of vibration (studied at the distal joints of the thumbs
and great  toes  using a  128  Hz  tuning fork) as  well  as routine laboratory
examinations,  including  peripheral hemograms,  determination of  blood  specific
gravity, and  urinalysis  for  urobilinogen and protein.   These  examinations
revealed no consistent  dose-related  adverse effects among the four groups of
workers.
     In the most recent  study,  Kramer et al.  (1978) measured numerous  physio-
logical parameters  of workers in  two adjacent  textile plants.   Detailed  blood
chemistry and hematology studies  were  conducted for 151  matched pairs  of
employees  to compare the exposed and unexposed partners.  All employees in the
exposed group  had been exposed  to MC (and  other  solvents),  in varying  concen-
trations,  for  up to 6 years.  The concentration  range was  11 to  838 ppm, with
a mean of 115 ppm MC.    It is questionable whether this period  of  exposure
would result  in  toxic  symptoms.  Because only healthy,  active  workers were
selected,  and the average length of exposure for the study population was less
002MC3/A                             5-9                              11-10-81

-------
than 1 year at the stated TWA,  no conclusions can be drawn about accumulative,
long term effects.   The control  group was only minimally exposed to nonchlori-
nated solvents.
     Pairs were matched  with regard  to age, race, sex, work shift, job des-
cription and  socioeconomic  status,  and  examined within a 10 week period.
Subject height, weight,  blood pressure and pulse were obtained, and electro-
cardiograms were  recorded.   Laboratory blood  determinations  included  hema-
tocrit, hemoglobin, red  blood cell count (RBC), white blood cell count (WBC),
mean corpuscular  hemoglobin (MCH),  mean  corpuscular volume  (MCV) alkaline
phosphatase, SGOT,  SGPT, gamma glutamyl transpeptidase, total bilirubin, urea
nitrogen,  LDH,  uric acid, total protein,  A/G  ratio, albumin, calcium, and
phosphorus.  For quantitative variables,  t tests and tests of homogeneity of
variables  were  made.   Multiple  regression analysis  was performed  on paired
differences with respect to environmental variables and on the combined matched
exposed and control populations with respect to demographic variables.
     Breathing  zone samples were collected in  charcoal  tubes, except in a few
locations  where area  sampling  was more practical, and analyzed on a portable
gas  chromatograph  equipped with  a flame  ionization detector.   Samples of
expired air were  analyzed  immediately  after collection  by gas chromatography.
     After explaining that some data were eliminated on the basis of subjects'
smoking habits, high  blood pressure, or prior illness, the authors presented
statistical findings  but no individual  data.   MC concentrations in the breath
ranged  from "less  than 5 ppm" to  "greater than  30  ppm,"  with  the majority
(127/151)  between  5 and 29 ppm.   Comparison of  the health test data between
exposed and control subjects revealed  no statistically  significant differences
except  in  SGPT and albumin.  These  differences  were not  discussed at  length
and  the authors concluded that no health  impairment was  suffered by workers
exposed to an average daily concentration  of 115  ppm MC.
002MC3/A                             5-10                              11-10-81

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     Nine women washing brass frames in open containers of pure MC were studied
from the  onset  of  their exposure and were the subject of a report in 1965 by
Weitbrecht.  Air measurements were made during the summer with methyl bromide
indicator tubes (of questionable validity for measuring MC).   An average of 10
ppm of MC was  found in the  general  room air and 20 ppm  in the worksite air.
In addition  to  vapor  exposures,  the women's hands  were immersed in liquid MC
for varying  periods of time.  Breathing zone samples  were not collected.   It
seems likely that  breathing  zone concentrations  would have been considerably
higher if the women were working directly over the  solvent containers.
     The  women  experienced  transient  irritation of  the conjunctiva and/or
upper respiratory  passages  and  a  characteristic burning  sensation  of the
tongue.   They also reported that their teeth felt dull (rhubarb effect).   They
did not experience  a  burning sensation or  initial swelling when their hands
were placed in liquid  MC; however,  they did have  a  feeling of ice-cold fingers.
A sustained  paleness  of the  fingers occurred only at the beginning of the
work, otherwise it appeared only when the exposure  was continuous.
     After the windows  were  closed  in the fall,  the women complained of loss
of appetite, pressure  sensation in  the stomach,  vomiting, tiredness,  headache,
and insomnia.  The author considered that these  complaints were neurotic chain
reactions influenced,  in part,  by safety signs in the  shop.   He did not differ-
entiate the  effects of  the MC exposure from the possible  suggestive effect of
the safety  signs, and  did  not comment upon  the lack of the safety sign effect
in the summer.
     Clinically, he found hypertension in six of  the women and positive urobi-
linogen in  two;  in addition,  he reported  what  he  described as autonomic
dystrophy  in  two,  circulatory  dystrophy in one, and  psychasthenia in one.
5.1.3  Accidental  Exposure
     Accidental  exposure to  MC  can  lead to death.   Table 5-4 lists the signs
and symptoms  of a number  of cases  in which the patients survived.   These
002MC3/A                             5-11                            11-10-a

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§
f\>
o
to
en
 i
                                      TARIE 5-1.   SIGNS AND SYMPTOMS Of PAIMNIS SURVIVING
                                                    INIOXJCrtUON WJfll rlfimri MHOHmORM
                Reference
                 Patient
                                              Amount
               BP*    imb   sr,Pic  SOOT     APP   BUN'
ECG"
                                                                                                                      Comments
              Stewart and       17 M
               Andrews, 19r«<>
              SlPwart, 1P71
                                44 WM
              Stewart. 1971     55 WM
              Stewart, 1971     47 WM
lilt and Cohen,   5 torn-
 19
-------
results suggest that MC  has  only a minimal potential for producing liver or


kidney injury in man.  The primary toxic effect appears  to  be a reversible


depression of the  CNS, typical  of an anesthetic agent.   In addition,  cardio-
                            \

vascular parameters may be altered by exposure to MC.


     Two  fatal  cases  in which  the  subjects  intentionally inhaled cleaning


fluids containing  MC were  reported by Hall and  Mine  (1966).   A 19-year-old


woman who was observed  sniffing cleaning fluid  over several days and acting


irrationally was later found  dead.   Pathologic findings on autopsy were con-


fined  to  the respiratory  system,  stomach,  and brain.  The  vessels of  the


bronchi were  congested,  and  the bronchi contained  thick, yellowish-brown


secretions.   There was  passive  congestion  throughout  the  lungs,  and  the


parenchyma  showed  considerable  amounts  of  thick, dark red  blood  and thin


frothy fluid in the congested areas.  The mucosa of the stomach was hyperemic;


the leptomeninges  were thin,  glistening, transparent and markedly congested;


brain ventricles contained clear cerebrospinal fluid; vascular markings were
                                «,

prominent,  and  there  was  acute", passive congestion  throughout the brain.


     There were indications  of  chronic,  intentional inhalation of a cleaning


fluid containing MC  in  the other fatal case studied by Hall and Hine  (1966).


On autopsy,  pathologic findings were confined to  the  respiratory  system and


the kidneys.  The  lungs were congested and edematous, the vessels dilated,  and


small  hemorrhagic  areas  were present.   The  kidneys showed marked  vascular


congestion around  the pyramids, especially on the periphery.


     In neither of these cases  were drugs or  solvents  detected in  the stomach


contents, and no barbiturates were  found in the blood. Blood levels of MC were


72.0 and 13.0 mg percent,  respectively.


     Twenty-nine cases of  sudden death from  sniffing  MC  during 1964  to 1969


were  reported  by  Bas (1970).   These were among 110  cases of  sudden  death





002MC3/A                             5-13                             11-10-81

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attributed to sniffing volatile hydrocarbons and halocarbons  summarized  by  the


author.  In  18  of  the 110 cases, death followed  sniffing  coupled with  some


form of exercise.   No anatomical abnormalities were found from gross  or  micro-


scopic post  mortem examinations that could explain the  sudden deaths.   The


author discussed the  possibility that resulted from cardiac  sensitization  to


endogenous catecholamine.


     Six fatal cases were reported by Stahl et al. (1969).   In the first case,


a 20-year-old man was  found dead in  a closed  space  in which he had been work-


ing with a  "paint  remover."   Gas chromatographic analysis revealed MC in all


tissues.  Upon  autopsy, the  lungs were found to  be congested and moderately


edamatous; the  liver,  spleen,  kidneys, and  brain  were also congested.  Micro-


scopic examination of the brain suggested anoxia as the cause of  death.


     In the  second  case,  a 17-year-old man was found dead in a room in which


he was  cleaning an air vent with MC.  Gas  chromatographic analysis  revealed


the presence of MC although blood levels were only 0.15 mg percent.  At autopsy,
                                i

the skin was moderately cyanotic, the brain, liver, kidneys, and spleen were


moderately congested,  and the  lungs were markedly edematous with evidence of


aspiration  of gastric contents.  Blood  lactic  acid levels were high.   The


authors concluded that death was probably due to 0^ deprivation.


     In the  third  case, an obese 24-year-old man was found dead in bed after


having  cleaned  electrical equipment  with  MC.   At necropsy, marked  cyanosis of


the head  and neck  were noted;  the  lungs  were congested and edematous.  The


brain,  spleen,  and kidneys were  also congested.


     In the  fourth,  fifth,  and sixth cases, three males were found dead in an


unventilated 4  x 5 ft.  compartment  where  they  had been cleaning  electrical


equipment  with  MC.  Autopsies  revealed  congestion and  edema of  the lungs.


Blood  levels of MC were 12.0 mg percent, 6.2  mg  percent, and  6.0 mg  percent  in


these  victims.


002MC3/A                              5-14                             11-10-81

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     Hatfield and Maykoski (1970) reported on a 27-year-old who was  found  dead



in an  aircraft  tank  that he had been cleaning  with  MC.   Upon autopsy, the



lungs  showed  some passive congestion and some edema.   The kidneys showed some



passsive congestion  and  there  appeared  to be some slight edema of the brain.



Microscopic examination  disclosed  a pulmonary  parenchyma  with considerable



acute  passive congestion and focal extravasations of  red blood cells  into



alveolar spaces.  Upon  section,  the  renal  parenchyma was intensely congested



and of  a  dark red color.  Microscopically, there was intense, acute passive



congestion.   There  appeared to  be  marked congestion of  the  small  vessels



within the white matter of the brain.   The diagnosis  was acute passive conges-



tion of the viscera with petechial  hemorrhages in the lung and brain.   Hatfield



and Maykoski  estimated the concentration level  of MC to which  this worker was



exposed at 62,000 ppm.



     Caplan et al.  (1976) reported a fatal  intoxication in which a 40-year-old



female was apparently overcome while painting a bathroom.  On external examina-
                               «.
                               4

tion,  the  only  significant  abnormality  was in  the respiratory system.   The



bronchi contained frothy fluid.   On section, the lungs exuded moderate aimounts



of frothy fluid.  Histologically, the lungs showed acute edema and congestion,



and the liver showed a mild fatty change.   MC was identified in tissue s.amples.



     Twenty-one deaths  from abuse or gross  misuse of decongestant aerosol



sprays containing MC  in  the solvent resulted in removal of several  such pro-



ducts from the market (Federal  Register 38:21935-36,  1973).



5.2  EFFECTS ON ANIMALS



5.2.1  Acute and Subacute Effects


     The LD   's Of MC  for various species are  found in Table 5-5.   Admini-



stration of single oral  doses  yielded LDcn'5  ^or  laDorat-ory  animals  ranging


from 8.6  gm/kg for guinea pigs  to  14.3 gm/kg  for rats  (Torkelson  et al.,






002MC3/A                             5-15                             11-10-81

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                     TABLE  5-5.  ACUTE TOXICITY OF METHYL CHLOROFORM
Reference
Torkelson et al. ,
1958
Torkelson et al. ,
1958
Torkelson et al. ,
1958
Torkelson et al. ,
1958
Torkelson et al. ,
1958
Takeuchi
1966
Priestly and Plaa
1976
Klaass'en and Plaa
1969
Torkelson et al. ,
1958
Adams et al. ,
1950
Plaa et al. ,
1958
Klaassen and Plaa
1967
Klaassen and Plaa
1967
Gehring
1968
Species
rats
rats
mice
albino
rabbits
guinea
pigs
SM mice
CF-1 Swiss
mice
Sprague-Oawley
rats
albino
rabbits
rats
Princeton
mice
Swiss-Webster
mice
dog
Swiss-Webster
mice
Route
oral
oral
oral
oral
oral
i.p.
i.p.
i.p.
dermal
inhalation
i.p.
i.p.
i.p.
i.p.
Sex
M
F
F
mixed
mixed
?
M
M
mixed
mixed
M
M
M
F
LD50
»g/kg
14,300
11,000
9,700
10,500
8,600
2,568
4,008
5,054
15,800
18.0001
16,000
5,080
4,140
4,700
95% confidence
limits
12,100 to 17,000
9,500 to 13,000

9,700 to 11,300
6,100 to 12,200

3,558 to 4,522
4,389 to 5,586


12,700 to 21,400
4,140 to 6,010

4,320 to 5,110
LC50 in ppm for 3 hr exposure;  7 hr exposure resulted in a LC,0 of 14,250 ppm (12,950 to
15,675).                                                      50
        002MC3/A
5-16
11-10-81

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1958).   The  solvent caused slight  transitory  irritation  in the eyes.   The

toxicity from  skin  absorption was  found to be  low, as doses of 4 gin/kg failed

to kill  any  rabbits exposed for 24 hrs.   Repeated multiple daily application

of 500 mg/kg  to the skin of rabbits  caused  no effect other than reversible

local  irritation.   In other experiments,  laboratory animals  were exposed

repeatedly to 500, 1000, 2000,  and 10,000 ppm in order to establish conditions

safe  for repeated exposure.  Rats, guinea pigs,  rabbits, and monkeys were

unaffected after  6  months  of repeated 7 hr  exposures  5 days/wk to 500 ppm.

Female guinea  pigs,  which  were found to be  the most sensitive in previous

experiments, were able  to  tolerate 1000 ppm for  0.6 hr/day for 3  months and

2000 ppm for 0.1 hr/day with no detectable adverse effects.  Male rats tolerated

exposure to 10,000 ppm for 0.5 hr per day with no organic injury.   The effects

of MC were  shown  to be primarily  anesthetic,  with only a slight capacity  to

cause reversible injury to the lungs and liver.  Based on this work, Torkelson

et al. (1958) suggested that the maximum allowable concentration for MC be 500
                               «,
ppm.

     In  another study,  Eben and Kimmerle (1974)  exposed  rats  acutely  (4 hr)

and  subacutely  (8 hr/day, 5  times/wk  for 3 months) to 220 and  440  ppm  and  200

ppm,  respectively.  During the periods of exposure, the animals did not differ

in any way  from their controls in behavior,  appearance, or body weight gain.

Hematologic examination performed  at  the end  of  the exposure  period did  not

reveal any pathologic abnormalities.  Liver and renal function tests and blood

glucose  were  normal.  At autopsy,  the organs of the  exposed rats were  normal;

organ weights did not deviate significantly from those of the control animals.

     Prendergast et  al.  (1967)  repeatedly exposed 15 Sprague-Dawley rats,  15

Hartley  guinea  pigs,  3  squirrel  monkeys, 3 New Zealand albino rabbits, and 2

beagle dogs (8 hr/day, 5 days/wk for 6 weeks) to 2200 ppm MC.  The same number



002MC3/A                             5-17                             11-10-81

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of animals were exposed continuously for 90 days to 135 ppm and 370 ppm.  The
repeated exposure to 2200 ppm did not result in any deaths or visible signs  of
toxicity, although weight  loss  was  observed in rabbits and  dogs.   The con-
tinuous exposure at  370  ppm did not cause  any  deaths,  visible toxic signs,
significant growth depression or biochemical,  hematologic  or pathologic changes.
Continuous exposure  to 135 ppm resulted in three deaths but no visible toxic
signs or  impaired  growth in any of  the survivors.   Autopsy and subsequent
histopathologic examination of  the experimental animals revealed lung conges-
tion and  pneumonitis which may have been  severe  enough to have caused the
deaths of the  two  rats and the  rabbit, but which were  not attributed to the
exposure.
     Because chlorinated  hydrocarbons  are  known to  produce a  specific  hepato-
toxicity, the  effects of  MC on  hepatic function have been studied  by a  number
of different investigators.  Lai and Shah (1970) exposed Swiss-albino, random-
bred male mice to  3000 ppm for either 24 hr or for 4 or 8 hr/day for several
days.   Hexobarbital  (80 mg/kg)1^,  barbital sodium  (275 mg/kg)- or chloral
hydrate  (350 mg/kg)-induced  sleeping time  was  measured  at predetermined times
after  exposure.   In  addition,  hexobarbital oxidizing  activity, nitro-group
reducing  activity,  and the protein content of  the  liver  supernatant fraction
were also determined.   Inhalation  of MC at a  concentration of 3000 ppm for  24
hr produced  a  maximum reduction in the duration of  hexobarbital sleeping  time
but had no effect on hypnosis produced  by  barbital  or chloral  hydrate.  Hypnosis
returned  to  the control  level  at 48 hr after  the  termination of  exposure.
Repeated  short exposures, 4 or  8  hr/day  for  several days,  had a  cumulative
effect, but a  single 8 hr  exposure did not.  The  supernatant  fraction  isolated
(9000  g)  from livers of  mice  previously  exposed to MC  oxidized hexobarbital
more  efficiently,  but the ability  to reduce p-nitrobenzoic  acid  remained

002MC3/A                              5-18                              11-10-81

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unaffected.   The protein content  of  this liver supernatant fraction  was  also
unaltered.
     Fuller et  al.  (1970)  made further observations on  Lai  and Shah's in-
vestigations of the  inductive  effect of MC on hepatic  drug Metabolism.   Male
Sprague-Dawley  rats  and Swiss-albino,  random-bred mice  were exposed to MC
(2500 to 3000 ppm)  in  a dynamic chamber for 24 hr.  Loss  of righting reflex
due to hexobarbital  (120 mg/kg), meprobamate (300 mg/kg),  or zoxazolamine (80
mg/kg) administration  and  iji  vitro activity of hexobarbital oxidase, zoxazo-
lamine hydroxylase,  aminopyrine demethylase,  reduced  nicotinamide adenine
dinucleotide phosphate  (NADPH)  cytochrome  c reductase, and CO-binding pigment
(cytochrome P-450) were then  measured.   Pretreatment with inhibitors of pro-
tein synthesis (cycloheximide and actinomycin D) were used to block the effect
of MC  on  the hepatic  drug-metabolizing  system.   Inhalation of MC (2500 to
3000 ppm) for 24 hr decreased the duration of action of hexobarbital, meproba-
mate, and zoxazolamine.  This was accompanied by an increase in the metabolism
                                *
of hexobarbital, zoxazolamine  find aminopyrine uj vitro by hepatic microsomal
enzymes.   Cytochrome P-450 and cytochrome c  reductase were also increased;
however,  liver  weight  and  liver microsomal protein were not enhanced.  Pre-
treatment of  rats  with protein synthesis  inhibitors prevented the MC induced
decrease in  hexobarbital sleeping  time  and the increase  in hepatic drug meta-
bolism.  The authors concluded that MC may be  inducing the production of a new
enzyme protein  and  that this protein may be used  as a  "prepathologic" measure
of MC toxicity.
     Savolainin et  al.  (1977)  investigated the effects of MC on rat  brain  and
liver after intermittent inhalation.   Ten  male Sprague-Dawley rats were exposed
to 500 ppm  MC for 4 days,  6  hr daily.   The right cerebral hemispheres were
homogenized and analyzed for protein,  RNA, glutathione, and for the activity

002MC3/A                             5-19                             11-10-81

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of acid proteinases.   Liver microsomes were analyzed for the amount of  protein

and for cytochrome P-450 content.   During exposure,  the behavior of the exposed

animals was analyzed  In an open field test and did not differ significantly

from that of  the  controls.   Brain protein content was not affected; however,

the amount of  RNA was lower than  in  the  controls.   MC decreased the  liver

microsomal cytochrome P-450 content after cumulative exposure to the material

for 5  days.   The  authors explained the decrease  in cytochrome  P-450 by the

previously observed (Pelkonen and  Vainio, 1975) affinity  of MC for  the  type I

binding site.

     McNutt et  al. (1975)  demonstrated hepatic lesions  in mice after continu-

ous inhalation  exposure to MC.  CF-1  mice were exposed  to 0, 250,  or 1000 ppm

MC continuously for  up  to 14 weeks with a weekly serial  sacrifice.  At 1000

ppm, from 1 to 14 weeks, significant changes in centrilobular hepatocytes were

observed.  Moderate  liver  triglyceride accumulation was  evident in the 1000

ppm group  and peaked at 40 mg/gm  of  tissue after 7 weeks of exposure  (by 14
                                v
weeks  the  triglyceride  level  hSd  not decreased to  16 gm).   Electron micro-

scopic  evaluation revealed that cytoplasmic alterations  were most severe in

centrilobular  hepatocytes  in  the  1000 ppm group  and were mild  to  few  in the

250 ppm group.  The  alterations consisted  of  vesticulation  of the  rough endo-

plasmic  reticulum, with loss  of attached polyribosomes,  and increased  smooth

endoplasrm'c reticulum,  microbodies,  and triglyceride droplets.   Necrosis of

individual hepatocytes  occurred in 40 percent of  the mice exposed  to 1000 ppm

for 12 weeks.   This  necrosis  was  associated with  an acute inflammatory infil-

trate  and hypertrophy of Kupffer cells.  This study concluded that  the  effects

of MC  were similar to carbon  tetrachloride,  but  they appeared  to be much less

severe.
 002MC3/A                             5-20                              11-10-81

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            TABLE 5-6.   THE RELATIVE HEPATOTOX1C EFFICACY OF
                          CHLORINATED SOLVENTS
                                        Relative hepatotoxic
     Compound                                efficacy


1,1,1-trichloroethane                            1


Tetrachloroethylene                              3


Trichloroethylene                                8


Sym-tetrachloroethane                           12


1,1,2-trichloroethane                           40


Chloroform                                      60


Carbon tetrachloride                           190


Adapted from Plaa et al., (1958).


     Table 5-6 shows  the relative hepatotoxic effect of MC  in comparison to
                               b

other chlorinated  solvents.  MC,  according  to  Plaa et al.,  (1958), was judged


to be the least hepatotoxic of the seven solvents investigated.


5.2.2  Central Nervous System Effects


     The  most  notable pharmacologic  effect of methyl chloroform following


pulmonary vapor  absorption is  its  action  upon  the  central  nervous  syteir,.


Table 5-7 shows  the occurrence of neurological  signs of toxicity associated


with progressively  increasing  inhalant  concentrations of MC  in a variety of


species.   At  higher than  anesthetic  concentrations  (approx.  10,000 pprc),


severe respiratory depression develops and cardiac arrest may occur.


5.2.3  Cardiovascular Effects


     The cardiovascular  effects produced by exposure to MC  have been exten-


sively studied in recent years.  Halogenated alkanes have been shown to sensitize


the heart to  catecholamines  and at the same time produce cardiac depression.


002MC3/A                             5-21                             11-10-81

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                  TABLE 5-7.   PROBABLE  RESULT  OF  SINGLE  EXPOSURE
                        TO THE VAPORS OF  METHYL CHLOROFORM
Exposure    Concentration
  time          in air              Expected effect  in  humans
  (min)         (ppm)


    5          20,000         Complete incoordination and helplessness  (R)
               10,000         Pronounced loss of coordination  (R)
                5,000         Definite incoordination (R, M)
                2,000         Disturbance of equilibrium.   Odor is  unpleasant
                                but tolerable (H)

   15          10,000         Pronounced loss of coordination  (R)
                2,000         Loss of equilbrium (H)
                1,000         Possible beginning loss of equilibrium (H)

   30          10,000         Pronounced loss of coordination  (R)
                5,000         Incoordination (R, M)
                2,000         Loss of equilibrium (H)
                1,000         Mild eye and nasal discomfort; possible
                                slight loss of equilibrium (H)

   60          20,000         Surgical anesthesia, possible death  (R)
               10,000         Pronounced loss of coordination  (R)
                5,000         Ofivious loss of coordination (R, M)
                2,000         Loss of coordination (H)
                1,000         Very slight loss of equilibrium  (H)
                  500         No detectable effect,  but odor is
                                obvious (R, H)
                  100         Apparent odor threshold (H)
(H) Expected effects are based on human data.
(M) Expected effects are based on monkey data.
(R) Expected effects are based on rat data.

From Stewart, (1968, 1971), Torkelson et al.  (1958), and Toxicology
Committee, Am. Ind. Hgy. Assoc., 1964.
 002MC3/A                             5-22                             11-10-81

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     Krantz et al.  (1959)  were the first to observe  the  cardiac depression
produced by MC.   Rats were deeply anesthetized with MC for 1 hr,  at which time
the hearts were immediately removed.   Cardiac ventricular  slices  were promptly
prepared  and  the oxygen  uptake  over the 1  hour  period was Measured.  MC
anesthesia was  associated with  a  significant  diminution  (33.3 percent)  in
oxygen uptake of the myocardium.   Blood pressure studies in dogs  also revealed
that anesthesia with MC elicited a depressed response.  At the point of respi-
ratory arrest, the  blood  pressure was reduced  to approximately  one-half  its
normal value.  Ether anesthesia  under similar  conditions results  in only a
slightly  depressed  blood  pressure.   Six dogs  and  two  rhesus  monkeys were
anesthetized  and  electrocardiograms  recorded.   The pattern  of the electro-
cardiogram was essentially  unaltered;  the heart rate was  increased and the
T-wave was either flattened or inverted.  At the point  of respiratory arrest,
a depressed S-T segment was observed; however, tachycardia was absent.
     Herd and co-workers  (1973,  1974) confirmed the  initial observations of
Krantz et al. (1959).  They  exposed  dogs to MC (104-4 x  10  ppm/min) and
observed  a dose-dependent  biphasic decline  in  arterial  blood pressure.   Since
cardiac output increased initially, the  initial decline in pressure (within 10
to  15  seconds after introduction of MC)  was  due  to a decrease in total
peripheral resistance  (TPR).   Injection  of phenylephrine (pure alphaagonist)
reversed  the  peripheral vascular effects,  indicating that  MC does  not  act
directly  on  the vascular musculature.   The  second phase  of blood  pressure
decline was  found  to be associated with a  decrease in  myocardial  contracti-
lity, reflected  by  a decline  in  both  heart  rate and stroke  volume.   Exogenous
Ca** reversed the  MC induced  decline in myocardial contractility, but had no
effect on the initial phase of peripheral vasodilation.
002MC3/A                             5-23                             11-10-81

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     Taylor et al.  (1976)  exposed  New Zealand white rabbits  to  a  series  of

haloalkane chemicals.  The animals were first anesthetized with sodium pento-

barbital, then fitted with  various  cannulas allowing for the measurement of

mean arterial  pressure,  left  ventricular  pressure,  left ventricle  dP/dt,

cardiac  output,  stroke  volume,  heart  rate, left ventricular end-diastolic

pressure,  central  venous  pressure,  and  peripheral  vascular  resistance.

Table 5-8 lists the left ventricular and hemodynamic  effects of 50,000 pprr MC.

There was  no  effect on  heart rate,  left ventricular end-diastolic pressure,

central  venous pressure, or peripheral vascular  resistance.  During exposure,

no cardiac arrhythmias were observed nor was there a significant change in pH,

oxygen,  or carbon  dioxide  tensions.   As a cardiac depressant, MC appeared to

be as potent as halothane and fluorocarbon 11.

     Numerous  hydrocarbon  compounds have  been  shown to sensitize the myo-

cardium  to catecholamines.   In humans, for example, several unexplained deaths

have been associated with solvent abuse or overexposure; ventricular fibrilla-
                               »
tion  due to  cardiac  sensitization has  been  suggested as  the underlying

mechanism.  The  cardiac  sensitization potential of MC  has  been  investigated

under  various  circumstances.   In an early  study,  Rennick  et al. (1949) re-

ported  MC and epinephrine  induction of  idioventricular  rhythms  in  dogs.

Attempts to  induce MC anesthesia resulted  in the  sudden death, presumably of

cardiac  origin,  of  two  animals.   Ventricular extrasystoles and ventricular

tachycardia were  seen in five dogs  under barbital  anesthesia when  epinephrine

was  injected  after "repeated  small doses"  of MC.   Maximum sensitization of the

heart  to epinephrine  occurred after  the  administration  of 0.25 to 0.4  ml/kg  of

MC.   Further administration  of  this compound  produced severe hypotension.

Somani  and Lum (1965),   Lucchesi (1965), and  Hermansen  (1970)  administered MC
 002MC3/A                             5-24                              11-10-81

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o
o
ro
3
n
OJ
ro
tn
                            TABLE 5-8.   LEFT VENTRICULAR AND HEMODYNAMIC EFFECTS OF METHYL  CHLOROFORM
a



Pre-exposure
1 minute post
exposure
Mean
Arterial
Pressure
(mmHg)
69±5
45+4
Left
Ventricular
Pressure
(mmHg)
97±3
7*0+5
Left
Ventricle
dP/dt
(mmHg/sec)
3792+325
1433+163

Cardiac
Output
(ml/min)
423+31
266+26

Stroke
Vo 1 ume
(ml)
1.47+0.09
0.92+0.07

 All values at 1 minute exposure are significantly different from pre-exposure  values  at p <0.05.


Source:   Taylor et al.  (1976).
 I
«—"
o

oo

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to dogs or  mice  as  a method for induction of cardiac arrhythmias  in studying
and characterizing  the sensitivity and  specificity  of adrenergic blocking
agents.
     Clark and Tinston  (1973)  evaluated  the cardiac  sensitizing potencies  of
14 halogenated  hydrocarbons in conscious beagle dogs.   During  the last 10
seconds of  a 5 min exposure, epinephrine  (5 mg/kg) was injected intravenously
(bolus).   As a control,  epinephrine was  given prior to exposure and also 10
min after the end of exposure.  The electrocardiogram (lead II) was monitored
continuously during this procedure.   Several dose levels  of the  chemical being
tested were used, each differing  by a factor of two.   The concentration at
which 50% (EC50) of the animals could be  sensitized was calculated by a moving
average interpolation.  The  mean  concentration  of  MC which produced cardiac
sensitization in 50%  of the animals tested was 7,500 ppm  (4,000 to 11,000).
The authors found that they could directly predict  the ECrQ for  cardiac sensi-
tization by knowing the vapor pressure at 37°C and  the partial pressure at the
EC^Q.   Table 5-9  lists the EC^Q for a number of different halogenated hydro-
carbons.   MC appears to be a highly potent sensitizing agent,  with tetrachloro-
difluoromethane being  the  most potent in the series.  The authors  concluded
that cardiac sensitization was  probably  a structurally nonspecific  action and
that it was physically toxic.
     Reinhardt et al.  (1973) investigated five commonly used industrial and
household solvents, including MC,  to assess their cardiac-sensitization poten-
tial.   The investigators conducted experiments on unanesthetized,  healthy male
beagle dogs that had been trained to breathe through a one-way face mask while
supported in a standing position  by a  sling.  The  dog  inhaled house air for  7
minutes and the test compound for 10 minutes.  Epinephrine (8 ug/kg in  1 ml of
normal saline) was injected  i.v.  in 9 seconds as a control dose after 2 minutes

002MC3/A                             5-26                             11-10-81

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                  TABLE  5-9.  CONCENTRATION OF CHEMICALS CAUSING CARDIAC
                   SENSITIZATION  AND THEIR PHYSICAL PROPERTIES (IN PPM)



hemical
etrachl orodi f 1 uoromethane
arbon tetrachloride
richloroethane
alothane
richlorotrif luoroethane
ethylene chloride
richlorof luoromethane
ichlorof luoromethane
ich-lorotetraf luoroethane
inyl chloride
ropane
romotr i f 1 uoromethane
hlorotrif luoromethane



EC50
1,200
5,000
7,500
20,000
10,000
24,000
12,500
25,000
100,000
50,000
200,000
200,000
800,000


Vapor pressure
at 37 C (mmHg)
Ps
99
190
210
480
524
661
1,186
2,052
2,310
4,218
9,538
15,276
40,698

Partial
pressure at
EC50(mmHg)
Pcs
2
4
6
15
8
18
10
19
76
38
153
153
610
Relative
saturation
for cardiac
sensitization
Pcs/Ps
0.02
0.02
0.03
0.03
0.02
0.03
0.01
0.01
0.03
0.01
0.02
0.01
0.02
ource:   Clark and Tinston (1973).
      002MC3/A
5-27
                                                                            11-10-81

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of breathing air,  and  as a challenge dose after  5  minutes  of  breathing the
test compound.   The  10-minute  interval  between the  two doses was found to be
adequate to prevent  additive effects when tested  in 13 control  dogs subjected
to the  same procedures,  but with air substituted for  the  test compound.   A
positive response  was  considered to be  either  cessation  of cardiac output
(ventricular fibrillation)  or  the  development of an arrhythmia that was not
observed following the control dose and  that was  considered  to pose a  serious
threat  to  life (multiple,  consecutive  beats of  ventricular origin).   The
concentration of the test compound being delivered to the animal was  deter-
mined by gas chromatography at 2-minute intervals.
     The results obtained for MC indicated no response in 12 dogs at a nominal
concentration of 0.25  percent  (V/V)  and a marked response  in  3 of 18 dogs
exposed at a nominal concentration of 0.5 percent (V/V); 12 of  12 responded at
a  nominal  concentration  of  1.0  percent (V/V).   However,  unlike the  other
substances, MC-induced ventricular fibrillation in  the animals  from the  1.0
percent  group  reverted  to  multiple  consecutive ventricular beats within  a
matter  of  seconds  and  eventually recovered to  a  normal cardiac  rhythm.   The
results  confirm earlier  reports that MC  is  capable of sensitizing the  dog
heart to epinephrine.
     Trochimowicz  and  co-workers (1976) were concerned with the problem of
haloalkane-induced  cardiac  sensitization in  patients  who  had  survived myo-
cardial  infarction.  They induced myocardial  infarction  in beagle dogs and
tested  for  cardiac sensitization to  epinephrine  after  exposure  to MC  (2500,
3700, and  5000 ppm).  Myocardial  infarction  failed  to  significantly lower the
threshold  for  cardiac  sensitization.   There was no  greater potential for
cardiac  sensitization  among dogs having recovered  from myocardial  infarction
as compared to  normal healthy  animals.

002MC3/A                             5-28                              11-10-81

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     At sufficiently high  levels  (in the thousands of ppm, for durations of


exposure of the  order  of minutes), MC can cause  significant  alterations in


cardiovascular function  in a variety of experimental animals, including mice,


rabbits, dogs, and  possibly  monkeys.   The concentration levels reported for


these effects are generally  more  than an order of magnitude greater than the


current TLV for  humans of 350 ppm  (0.035 percent) and are  likely to be lethal


to the experimental  animals if the exposure is continued for more than several


minutes.  Indeed, some experimenters appeared to find it necessary to increase


the oxygen content  of  the inspired gas/air mixture to 40  percent to prevent


asphyxia of the exposed animals.  That they appeared unable to find effects at


lower exposure levels is noteworthy.  Also noteworthy is the fact that many of


the investigators reported complete recovery of the animals  if the exposure


was stopped.  This  was  the case,   for example,  with  the dog experiments of


Reinhardt et  al. (1973),  and Trochimowicz et  al.  (1976).   At  necropsy follow-


ing sacrifice of several  of  the animals several months after exposure,  there
                                t

was no  pathology attributable t*o the exposure per se.  Delayed cardiotoxicity


from acute exposure has not yet been demonstrated or ruled out.


5.3  TERATOGENICITY, MUTAGENICITY, AND CARCINOGENICITY


     It has been proposed that  the susceptibility of individuals to cancer  is


related to their ability  to metabolize carcinogenic compounds to benign  mole-


cules and, conversely,  to their capacity to  metabolize  benign  compounds to


carcinogens (Kellerman et al.,  1973).  Susceptibility may  be  partly explained


by genetic differences  involving  enzymatic pathways, repair  mechanisms, and


immune response mechanisms.  An assessment of the carcinogenic potential of MC


must therefore take into account   tissue concentrations  of MC during  typical


exposures (e.g., TLV-TWA,  350  ppm), as well  as the  extent and nature of MC
002MC3/A                             5-29                             11-10-81

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metabolism.   Both these  aspects  have  been extensively reviewed in Chapter  4.

While our knowledge is still limited in these areas, it can be concluded that

MC is metabolized  in  humans to a very small extent.  About 6 percent of the

body dose is  converted  by a hepatic microsomal oxidase system to trichloro-

ethanol.  The mechanism  of  this overall reaction is unknown, as is the poten-

tial for generation of "reactive intermediate compounds" which may contribute

to organ  toxicity and/or carcinogenesis.   Until  further  research is done,

particularly studies  to  determine  how and whether intermediate reactive com-

pounds  from MC  metabolism bind to cellular macromolecules, an evaluation  of

mutagenicity and carcinogenicity roust be made solely on the biological effects

of MC  in  various direct mutagen and carcinogen tests.  Unfortunately, MC has

not  been  subjected  to extensive and thorough testing.  The available studies

are  summarized  in Table  5-10.

5.3.1   Teratogenicity, Embryotoxicity,and Reproductive Effects

5.3.1.1 Overview—This  review subscribes to  the basic  viewpoints and defini-

tions of  the  terms "teratogeni£" and "fetotoxic" as summarized and stated by

Chernoff (1980):

     Generally,  the  term "teratogenic"  is defined as the tendency to produce
physical and/or functional  defects in offspring n\ utero.  The term "fetotoxic"
has  traditionally  been  used to describe  a wide  variety of embryonic and/or
fetal divergences  from  the normal which  cannot be classified  as  gross terata
(birth  defects) -- or which are of unknown or doubtful  significance.  Types of
effects which  fall under the  very  broad category of fetotoxic effects  are
death,  reductions  in fetal  weight,  enlarged renal  pelvis  edema, and increased
incidence of  supernumary ribs.  It should  be  emphasized,  however, that the
phenomena of  terata and fetal  toxicity  as currently defined are not separable
into precise  categories.   Rather,  the spectrum of  adverse embryonic/fetal
effects is  continuous,  and all deviations  from the normal  must be considered
as examples  of developmental  toxicity.    Gross morphological terata represent
but  one aspect  of this spectrum, and while  the significance of such structural
changes is  more  readily evaluated,  such effects  are not necessarily more
serious than  certain effects  which are  ordinarily classified as  fetotoxic--
fetal death being  the most  obvious example.

      In view  of the spectrum of effects at issue,  the Agency suggests that it
might  be  useful to consider developmental  toxicity in terms  of  three basic
subcategories.   The  first  subcategory  would be embryo or  fetal  lethality.


002MC3/A                              5-30                            11-10-81

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                                  TABLE  5-10.   ONCOGENIC TERATOGENIC TESTING OF METHYL CHLOROFORM
  Test System
     Species
     Reference
  Results
Teratogem'city

  Organogenesis


  Mammalian gestation

Carcinogenicity

  NCI bioassay - 2 yr




  NTP bioassay - 2 yr

  Industry bioassay -
    1.5 yr
Chick embryo
S/0 rat, S-W mice
Elovaara et al., 1979
Schwetz et al., 1975
Osborne-mendel rats, and'   NCI, 1977; NCI Clearinghouse on
  BfiC3F| mice and oral       Environmental Carcinogens, 1977
  gavage; 750 and 1500
  mg/kg (rat)

Rat and mouse gavage

S/D rats, inhalation 875
  and 1750 ppm
Positive; high toxic dose,
skeletal abnormalities

Negative
                                       Inconclusive; high animal
                                       mortality
                                       In progress

                                       Negative
NOTE:   See appendix for discussion of chemical  purity.

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This is,  of  course,  an irreversible effect and may occur with or without the
occurrence of gross terata.  The second subcategory would be teratogenesis and
would encompass those changes (structural and/or functional) which are induced
prenatally,  and  which are Irreversible.   Teratogenesis  includes structural
defects apparent  in  the  fetus,  functional deficits which nay become apparent
only after birth,  and any other long-term effects  (such as carcinogenicity)
which are attributable to ui utero exposure.   The  third category would be
embryo or fetal  toxicity as  comprised of those effects which are potentially
reversible.  This  subcategory would therefore include such effects as weight
reductions,  reduction  in the degree of skeletal ossification, and  delays  in
organ maturation.

     Two  major  problems  with a definitional  scheme of this nature must be
pointed out,  however.   The first is that the reversibility of any phenomenon
is extremely difficult to prove.   An organ  such as the  kidney,  for example,
may be  delayed  in development and then  appear to "catch  up".   Unless  a  series
of specific  kidney function  tests are  performed on the  neonate,  however,  no
conclusion may  be drawn concerning permanent organ function changes.  This
same uncertainty  as  to possible long-lasting after  effects  from  developmental
deviations is true for all examples of fetotoxicity.  The  second problem  is
that the  reversible  nature of an  embryonic/fetal effect  is  one species  might,
under a given agent, react in  another  species  in a more serious and  irrever-
sible manner.

     It is not possible, on the basis of  limited available data, to define the

full potential  of MC to produce adverse  teratogenic or  reproductive  effects.

Human epidemiology studies have not been conducted to evaluate the  effects of

MC  on  the exposed population.  'The available mammalian  studies were  not pro-

perly designed to  evaluate the  ability  of MC to produce  a teratogenic response

over a  wide  range of doses,  which should include doses high enough to produce

signs of  maternal toxicity and lower doses  which  do not produce  this effect.

The  teratology  studies which were performed with laboratory animals used rats

and mice  and only single doses  of MC which produced signs of  maternal toxicity.

Other studies in  chicken embryos  have  indicated that MC  disrupts embryogenesis

in  a  dose-related manner.  However,  since administration of  MC  directly into

the  air space of chicken embryo is not comparable to administration  of a dose

to  animals with a placenta,  it is not possible to interpret this  result in

relationship to  the  potential of  MC to  cause adverse human  reproductive effects.
 002MC3/A                             5-32                             11-10-81

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     In summary,  although  several  studies have been conducted to evaluate the
ability of  MC to  cause adverse teratogenic, embryotoxic  and reproductive
effects,  the  limitations  of  the available  data  does not  allow for  a full
assessment of  these  effects.   A better assessment of these effects could be
performed if the available studies met criteria similar to those suggested for
teratogenicity and reproductive testing (U.S. EPA, 1978).
5.3.1.2   Human Studies—No clinical  reports associate  maternal exposure to
methyl chloroform  with  congenital  malformations  in offspring.  No epidemic-
logical studies have been performed.
5.3.1.3   Animal Studies—All  studies  performed  to date in mammals have  been
done in rats and mice.  On the basis of these studies, it does not appear that
short- or long-term  exposure  to MC results  in teratogenic  effects in  rats, or
that short-term exposure  to  MC results in  such  effects in mice.   Delays in
fetal development have been observed in both species, but this may be a rever-
sible effect.  It  should be noted, however,  that  these  studies evaluated only
one dosage  level  per animal spe'cies, which  included dosages twice the maximum
excursion limit for  short-term exposures  of rats  and mice and six times the
maximum excursion  limit for  long-term exposures of rats.  Another  study in
chicken embryos  indicates that MC may have teratogenic  potential  in this
species.   However, since  it  is not known how comparative  studies in  chicken
embryos are to those in other mammalian species,  the teratogenic potential of
MC in chickens cannot be related to other animals without additional data.   In
addition,  no multi-generational studies of  mammalian reproductive performance
have been performed.
5.3.1.3.1  Rats--Schwetz et al. (1975) report results from Sprague-Dawley rats
exposed via inhalation to 875 ppm of MC for 7 hours daily on days  6 through  15
of gestation (Day 0 = the day sperm were observed in smears of vaginal contents).

002MC3/A                             5-33                             11-10-81

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Control  rats  were exposed  to  filtered air.   Dams were evaluated  for  body



weight gain,  food  consumption  and various organ weights.   Maternal carboxy-



hemoglobin level determinations were performed on blood samples collected via



orbital  sinus puncture  immediately  following the third and tenth (last) ex-



posure.  One-half  of the fetuses  in each  litter were examined for soft-tissue



malformations (free-hand sectioning),  and one-haIf  were stained and examined



for skeletal  malformations.  One  fetus in each  litter was randomly selected



and  evaluated using histological  techniques following serial  sectioning.



     Twenty-three  litters  from  dams  exposed  to 875 ppm of methyl chloroform



were evaluated.  No  effect was observed on maternal body weight or food con-



sumption.  The  mean absolute  liver  weight was  increased  as  compared  with



control, however the mean relative liver weight was unchanged.  No embryotoxic



or teratogenic  effects  were observed which were  attributable to maternal MC



exposure.



     York et  al.  (1981) exposed Long-Evans  rats  by inhalation  to  dosages of



2100 ± 200 ppm MC  for 6 hours da'ily, 5 days per week, in the following regime:



(1) two  weeks prior to mating through day 20 of gestation; (2) two weeks prior



to mating only; (3) throughout gestation only; (4) controls which were exposed



to filtered air before and during pregnancy.   Day 1 of pregnancy was designated



as the day  spermatozoa  were observed  in smears  of vaginal  contents.  One-half



of each  group was sacrificed on  day 21  and  assessed for signs of  maternal



toxicity, embryotoxicity or teratogenicity.   The other one-half were  allowed



to deliver young  naturally.  When possible, litters were culled to four pups



of each  sex  on day 4 postparturition  and to two pups  of  each sex  at 21 days



postpartum.   Pups  were  evaluated  for  behavioral  effects (open field activity,



running  wheel activity, amphetamine challenge) and  carcinogenicity.  Surviving



rats were sacrificed and necropsied at  12 months  of  age.







002MC3/A                            5-34                              11-10-81

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     York et al.  (1981)  reported that both total fetal body weight and male


fetal body  weight were  significantly depressed in  litters  exposed  during


pregnancy.   Delayed ossification was  more frequent in fetuses which  had been


exposed before and  during  pregnancy,  and a delay  in the  development of the


kidney was  a more  frequent  observation in fetuses  in  all  exposed groups.


These effects are  thought  to be indicative of  a slight, but possibly rever-


sible delay  in  development.    No treatment-related behavioral effects were


observed in the pups, nor were significant signs of maternal toxicity observed.


5.3.1.3.2  Mice--Schwetz et  al.  (1975)  exposed  Swiss-Webster mice to 875 ppm


of MC, twice the maximum excursion limit, for 7 hours daily, on days  6 through


15 of gestation (day 0 = day a vaginal plug was observed).   Dams were Caesarean


sectioned on day 18 of gestation and thirteen litters were evaluated.  Methodo-


logy  similar to  that described previously (Schwetz  et  al.  1979)  in  rats was


used, with the exception that food consumption was not monitored.


     Mean and relative maternal liver weights in the exposed rats were slightly
                                4.

but  not  statistically  decreased.   Also, fetuses from the exposed group were


slightly but not  statistically smaller (both the  crown-rump length  and the


body  weight).  This type of  observation is thought  to indicate a slight, but


perhaps  reversible  delay in  fetal  development.  No  increased  soft tissue  or


skeletal variations  of  the fetuses were attributable to maternal exposure to


MC.


5.3.1.3.3  Chicken  embryos—Elovaara  et al.  (1979)  evaluated the  effects  of a


number  of  aliphatic hydrocarbons  on the development of  chick embryos and


reported the toxicity and teratogenic potential of MC in relationship to those


of other aliphatic  chlorinated hydrocarbons.   MC was  injected into  the air


space of fertilized chicken  eggs at  2,  3,  and 6 days of incubation and the


rate  of survival or death after 14 days of incubation was used as a measure of






002MC3/A                              5-35                             11-10-81

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embryotoxicity.  The approximate  LD5Q  value  of MC varied between 50 and 100
uM/egg, depending upon time of injection.   The types  of abnormalities  reported
were absence of eye(s), upper beak deformations, exencephaly,  brain hemorrhages,
paleness (anemia), profound  edemas  either in the rump  sacs or  other  areas,
defects in  external viscera  and musculoskeletal defects of lower extremities.
The spontaneous background level  of malformations in  vehicle-injected  controls
included only  eye  and  skeletal  abnormalities.  This  study indicates that MC
has possible teratogenic potential when evaluated in  chicken embryos,  and that
additional  experiments should be performed for further evaluation.   A  scienti-
fic basis for  the  comparison of developmental  toxic effects in various animal
species is  needed before  results in  chickens  can be compared  with other
mammalian animal species.
5.3.2  Mutagenicity
     Methyl chloroform  has been tested for its  mutagenic potential  in several
systems (bacteria, yeast, and rats), but sufficient data are  not presented for
evaluating the results presented  in three papers (the Dow 1978 rat cytogenetic
study, the  Henschler  et al.  1977 testing  in  Salmonclla, and  the Loprieno el
al. 1980 testing  in  yeast).   Of the remaining papers, only two  described the
results of  mutagenicity testing  conducted in such a  manner  as  to prevent
evaporation of  MC  and  to ensure exposure  of  the indicator organisms.  Both of
these  tests (Simmon et  al.  1977 and  Snow et al.  1979) were conducted  in
Salmonclla and a weak positive response was reported for both.
     On the basis  of  the available  evidence,  it is concluded  that  MC  is  muta-
genic  in bacteria.  If the metabolism  and pharmacokinetics of this compound in
humans results  in  metabolic  products  which can  interact  with DNA as is the
case for bacteria, it may cause mutagenic effects in humans as well.  However,
additional  testing in other  organisms  (e.g.,  mammalian cells in culture)  is

002MC3/A                             5-36                             11-10-81

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necessary to confirm the mutagenicity observed in bacteria.   Careful  attention

should be given  to  the design and conduct  of  these  studies to prevent the

evaporation of MC, overcome its low solubility in aqueous media,  and  to ensure

exposure of the indicator organisms.

     Results from a joint  study sponsored by  Imperial Chemical Industries and

the Medical Research Council  in the  United  Kingdom and the  National  Institute

of Environmental Health Sciences in the USA, conducted to assess  the  ability of

short-term  tests  to predict  carcinogenicity,  have  recently been published

(8/6/81).   EPA  has  not  received a  copy of this report to  evaluate these data.

Although preliminary indications  are that MC tested negative, no information

was available  concerning which,  if  any, precautions were  taken  to  ensure

exposure of the organisms to the compound in these preliminary reports.  These

data will be  evaluated  critically  in the  near  future when the Agency receives

a copy of the final report.

     Price  et  al. (1978) exposed  Fischer  rat embryo cell  cultures  (F1706,
                               •.                                             i
subculture 108) to 1,1,1-trichlbroethane liquid at concentrations of  9.9 x 10

and 9.9  x 10  uM for 48 hours.  1,1,1-trichloroethane was diluted with growth

medium  to  yield  the appropriate  doses.   The  1,1,1-trichloroethane  sample

obtained from the Fisher Scientific Company was  purportedly >  99.9  percent

pure,  but  a personal  communication  from Carlson of  the Fisher  Scientific

Company  revealed that  methyl chloroform  supplied by  them to Dr. Price was

really the product of the Dow Chemical Company which is about 95 percent pure.

The cells  were  grown  in Eagles minimum  essential medium in Earle's  salts

supplemented  with 10  percent fetal  bovine  serum,  2  mM L-glutamine,  0.1  mM

nonessential amino  acids,  100 ug pencillin,  and  100  ug  streptomycin per  ml.

Quadruplicate cultures  were  treated  at 50  percent confluency with each dose.
002MC3/A                             5-37                             11-10-81

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After treatment, cells were  cultured  in growth medium alone  at  37°C.   Trans-
formation of cells treated with either dose level  of 1,1,1-trichloroethane was
observed by 23  and  44 days of incubation  and  was  characterized by  progres-
sively growing  foci composed of cells lacking contact inhibition and orienta-
tion.  There was  no transformation of cells grown  in medium alone or in the
presence of a 1:100 acetone concentration.  Fifty-two and 55 microscopic foci
per three dishes  with low and high 1,1,1-trichloroethane dose,  respectively,
were found  in dishes  inoculated with 50,000 cells from cultures treated four
subcultures earlier and  held for four weeks at  37°C  in a humidified CO^ incu-
bator prior to  staining.
5.3.3  Carcinogenicity
     [NOTE:  Refer  to Carcinogen Assessment Group's assessment  of MC attached
             to this document as Appendix  I.]
     At  this time,  direct animal  bioassays for MC  carcinogenicity  have been
carried  out  but are inconclusive.   Table  5-11  gives  the results of National
Cancer Institute (NCI) animal bioassays of congeners of MC.  The isomer 1,1,2-
trichloroethane is  carcinogenic in mice, inducing liver cancer and pheochromo-
cytoma  in  both  sexes.  Oi- and tetrachloroethanes  and hexachloroethane  also
produce  liver carcinomas  in mice.
     The initial  NCI  bioassay of MC  (1977)  was carried out  in  both sexes  of
Osborne-Mendel  rats and  BgC.Fj mice by oral gavage at two dose  levels (rats,
750  and 1,500  mg/kg,  5  days/wk;  mice, 2,807  and 5,615 mg/kg  time weighed
average) for 78 weeks.   Animals were observed  for another  12 weeks  for a total
of  90  weeks in the experiment.   In male rats, 64 percent given the low dose
and  72 percent  given the  high  dose died within the first  year,  while  48 per-
cent of  females given the low dose and 42 percent  given  the high  dose  died
 002MC3/A                             5-38                             11-10-81

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o
o
ro
3
O
TABLE 5-11.   SUMMARY OF NCI  CHLOROETHANE  BIOASSAY RESULTS AS OF JULY 1978 (Parker,  et al.,  1979)
        Compound
                             Species/sex    Tumor site
                     Statistically significant tumors
        monochloroethane
                             no testing planned
        1,1-dichloroethane
                                 retesting recommended because initial  results inconclusive
        1,2-dichloroethane
                             rats/female
                             rats/male
                                      mice/female


                                      mice/male
mammary gland
forestomach
circulatory system
subcutaneous tissue
mammary gland
endometrium
                                           lungs
adenocarcinomas
squamous cell carcinomas
hemangi ocarci nomas
fibromas
adenocarcinomas
stomal sarcomas
adenomas
adenomas
        1,1,1-trichloroethane
                                 retesting in progress
        1,1,2-trichloroethane
                             mice/female
                             mice/male
                             mice
1 iver
1 iver
adrenal glands
hepatocellular carcinomas
hepatocellular carcinomas
pheoc h romocy toma s
        1,1,1,2-tetrachloroethane
                                 testing in progress, no report available
        1,1,2,2,-tetrachloroethane
                             mice/male
                             mice/male
liver
liver
hepatocellular carcinomas
hepatocellular carcinomas
        pentachloroethane
                                 testing in progress, no report available
 I
 I—I
 o
 oo
        hexachloroethane
                             mice/female   liver
                             mice/male     liver
                        hepatocellular carcinomas
                        hepatocellular carcinomas

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within a year.   Similar mortality was observed in mice.  The high mortality
and symptoms of  other toxic effects indicate that the maximum tolerated dose
may have been exceeded.  These high doses may also alter metabolism.
     However, mortality was  also  high in untreated control  rats (30 percent)
and control  mice  (50  percent).   It should be noted that MC bioassay animals
were housed with animals receiving a variety of other chlorinated hydrocarbons.
Although a  variety  of neoplasms were found  in both treated and control rats
and mice (Table 5-12), there was no dose-related increased incidence of tumors
in either  sex  or species.   The NCI Report (1977) does not comment on whether
the failure  to  demonstrate  careinogenicity for MC is  due  to  lack of carci-
nogenic potential for this compound or due to the bioassay design and obviously
poor experimental conditions.  NCI is currently testing MC in  rats and mice by
gavage at lower doses.
     Table 5-13 summarizes  the  results  of an industry sponsored chronic bio-
assay  (Quast et  al.,  1978;  Rampy et al., 1977) by inhalation  exposure to MC.
Male  and  female rats were  exposed  6 hours/day,  5 days/week to 875 ppm and
1,750 ppm over  a  period of  52 weeks or  about  one-half their  lifetime.  The
actual amount  of  MC absorbed via the lungs by the rat and by  the tissues was
not determined  and  cannot be  estimated without metabolism and pharmacokinetic
data for the rat.  Based on the tumor incidence in the treated rats, which was
similar in classification to those of the control animals, MC  exposure did not
result in  a carcinogenic  outcome.  On the other hand, the inhalation dose of
MC was only two-  to threefold  higher than that  allowed  in  the  workplace  and
only one species was tested.
     In recent  years,  a great deal of attention has been focused on the  role
of MC  in the destruction of stratospheric ozone (0,).  Because 03 absorbs most
of the ultraviolet (UV) radiation in the 290 to 320 nm wavelength range before

002MC3/A                     '        5-40                              11-10-81

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o
o
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to
3>
TABLE 5-12. SUMMARY OF NEOPLASMS IN RATS AND MICE INGESTING
METHYL CHLOROFORM


Number of
Species Sex Animals
Rat Male 20
50
50
Female 20
50
50
in
i. Mouse Male 20
- 50
50
Female 20
50
50

3
Dose3
_
750
1500
_
750
1500

-
2807
5615
-
2807
5615

Total number
of tumors
3
6
4
14
6
12

5
2
9
5
2
3

Liver,
spleen
1
1
-
.
-'
1

2
1
8
2
1
"
Number of tumors observed

Lung
_
-
-
_
-
-

1
1
1
.
-
1
Kidney, Integumentary
bladder system
1
-
1
_ -
-
1

-
-
-
2
1
1
Heart,
vasculature
_
1
1
_
-
1

-
-
-
-
-
™
Brain,
pituitary
—
1
-
3
2
1

-
-
-
1
-
™

Other
1
3
3
11
4
8

2
-
—
5
-
1
      Compound administered in corn oil  by  stomach  tube  five  days per/week.
      Concentration is a time-weighted average  expressed in mg/kg/day.

     Source:   National Cancer Institute, 1977.
o
oo

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o
o
ro
                                             TABLE 5-13.  SUMMARY OF NEOPLASMS  IN RATS INHALING

                                                              METHYL CHLOROFORM
en
i
rvj

Number of tumors
Number of
Sex Animals
Male 189
94
92
Female 189
94
92
Concentration
(PPM)

875
1750
_
875
1750
Total number
of tumors
200
77
103
561
264
300
Liver,
spleen
2
1
-
\
1
3
Kidney,
Lung bladder
1 2
2
1
1 2
1
1
Integumentary
system
36
11
21
328
177
223
observed
Heart,
vasculature

-
1
2
-


Brain,
putuitary
39
13
21
102
28
35


Other
120
50
60
124
57
38

          Source:   Quast et al., 1978.
CO

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it reaches the earth's surface, a decrease in 0- concentration could be dele-
terious to biological processes.  It is known that nucleic acids and proteins
absorb significantly in this wavelength range, with molecular disruption that
results in genetic mutations.   Alteration of nucleic acids by UV irradiation
of tissues leads to an increased prevalence of skin cancer (National Research
Council 1976, 1979).
     Depletion of stratospheric 0,  levels have been correlated  with  an  in-
crease in the incidence of skin cancer (National  Research Council,  1976;  1979;
1982).  Accelerated usage of MC would be  expected to cause a further increase
in O^  depletion, thus resulting in higher incidence of skin cancer.  Prelimi-
nary  information from an ongoing epidemiological study of  skin cancer suggest
that the incidence of nonmelanoma skin cancer in the United States  may even be
greater (NCI, unpublished results,  August 1979).
5.4  SUMMARY OF  ADVERSE  HEALTH EFFECTS AND  LOWEST  OBSERVED EFFECTS LEVELS
5.4.1  Inhalation Exposure
     The high volatility of methyl chloroform  (MC), the extensive  use of this
compound as an industrial solvent,  and the early interest in MC as  an anesthetic
has resulted in many reports in the scientific literature of human  exposure to
MC.  Human experimental studies have been performed using acute and subchronic
exposures to MC.  Although  these  studies provide precise  information on ex-
posure levels, they  are  limited by the small numbers of subjects employed in
the studies  and  the  restricted number of physiological and behavioral para-
meters which were  assessed.   While information on  human  chronic inhalation
exposure to  MC  is  available from studies  of  occupationally  exposed groups,
there is uncertainty in these studies as  to the exact extent of exposure; some
studies are  complicated  by  simultaneous exposure to other chemicals used  in
the workplace.   Although these  human studies  have deficiencies which preclude

002MC3/A                             5-43                             11-10-81

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any individual  study  from being used for human risk assessment,  the combined
information presented  in  these  studies  does provide dose-response relation-
ships which give  a  relatively clear description of the toxic effects of MC.
Studies in experimental animals are available for acute and  subchronic  ex-
posure to MC, but the  uncertainty of extrapolation from animals to humans and
the relatively  large  number of controlled  human  studies  of acute exposure
makes this animal data useful only in a supportive role for human risk assess-
ment.   No adequate  chronic  studies  using experimental animals are available
and although  the reports  of human  occupational experience  with  MC are not
ideal, they provide the best information on the effects of chronic exposure to
MC.
5.4.1.1  Effects of Single Exposures—Lethal effects  of  acute exposure to MC
have been reported  after  accidental exposure and abuse,  with death resulting
from CNS depression followed by respiratory and cardiac failure (Hall and Hine
1966; Bas, 1970;  Stahl et al., 1969; Bonventre et  al.,  1977; Klienfeld and
Feiner, 1966; Halfield and  MayKoski,  1970; Caplan et  al.,  1976).   From the
levels of MC  in the blood and brain of cadavers,  estimates have been made of
the concentration of  MC present in the  air during  the time the victim was
exposed.  These  estimates range from 200 to 102,900 ppm, with  the  majority  of
values being  between  6,700 and  18,600 ppm (Table  5-14).  However,  these esti-
mates  are  extremely crude and  probably  low since it  is not  known to what
degree the MC had dissipated from the victims' body prior  to  obtaining tissue
samples.  Also,  some  of the observed deaths which occurred at relatively low
MC concentrations may have resulted from aspiration pneumonia  prior to exten-
sive  involvement  of the CNS.   A clearer  indication of the  levels  of MC needed
to produce narcosis is available in the study of Dornette and Jones (1960)
investigating the use of  MC as an  anesthetic  in humans.   Although  this study

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                 TABLE 5-14.  HUMAN FATALITIES ASSOCIATED WITH METHYL CHLOROFORM

Case
No. Sex
1
2
3
4
5
6
7
8
9
10
11
12
F
M
M
M
M
M
M
M
M
M
F
M
Material or
Product Inhaled
Energine
1,1,1-trichloro-
ethane
"degreaser"
K)
Chlorothene
1,1,1-trichloro-
ethane
n
11
"
"
"paint remover"
"paint thinner"
1,1,1-trichloro-
ethane
1,1
,1-Trichloroe thane
Concentrations
Lungs at Autopsy Blood mg%
passive congestion
no autopsy
acute congestion
and edema
heavy, edematous
heavy, edematous
heavy, edematous
heavy, edematous
heavy, edematous
moderately edematous
moderately edematous
acute edema & con-
gestion
markedly edematous
72

30.0
13
12.0
6.3*
6.2
6.0
6.0
4.7*
2.0
0.15
Air ppmx Remarks
102,900
75,000 ppm in work
space; collapsed
after leaving work
area
42,900
18,600 possible aspira-
tion, pneumonia
17,000
9,000
8,900
8,600
8,600
6,700
2,900 ethanol cone, in
blood was 0.04%
200 possible aspira-
tion, pneumonia
Source
Hall and Hine
Kleinfeld and
Bonventre et
Hall and Hine
Stahl et al. ,
Stahl et al. ,
Stahl et al. ,
Stahl et al. ,

, 1966
Feiner, 1966
al., 1977
, 1966
1969
1969
1969
1969
Hatfield and Maykoski, 1970
Stahl et al. ,
Caplan et al.
Stahl et al. ,
1969
, 1976
1969
 Calculations based on assumption  from  Stfwart et  al.,  1969,  of  one  hour  exposure correlating to 0.07  mg
 percent in blood per 100 ppm in air.
*flstimated from brain concentration.

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study gives an  indication  of  the levels of MC which  cause  CNS depression,



presumably a higher  concentration would be needed to  cause  depression  severe



enough to result  in  respiratory  or  cardiac failure.   From these  data and  the



LC50 concentrations of 14,250  ppm obtained by  Adams et al. (1950) for rats,  it



can be estimated  that  a single exposure to concentrations  of  MC less  than



5,000 ppm  are  probably  not potentially life threatening in humans, while



higher concentrations may produce narcosis and possibly death.



     The effects  of  exposure  of  human volunteers to MC for  single short  term



periods were reported  for MC  levels far below the  levels which cause narcosis



(Table 5-15).    Stewart  et al.   (1961) exposed 7 subjects to vapors of MC which



were increased  from  0  to 2,600 ppm in  increments  of  15 minutes  each.   The



progress of observed  effects  is  presented in Table 5-1, with 6 of 7 subjects



complaining of  throat  irritation at 1,900 ppm and  lightheadedness and incoor-



dination occurring  between 2,600 and  2,650 ppm.   The progressive exposure



method used in  this  study  and the small number of subjects  makes the  results



difficult to interpret.  However, in  a second experiment in which  subjects



were exposed to  900  MC for periods  of  20, 35, or  73 minutes, difficulty was



observed in the performance of the Romberg tests, and  some lightheadedness was



experienced.   No  effects were observed in subjects exposed  to 500 ppm  for MC



for 78 or 186 min.  In both of these experiments, commercial grade MC  was used



which contained  the inhibitors dioxane (2.4-3.0%), butanol  (0.12-0.3%)  and



small amounts of  1,2-dichloroethane.   These compounds may have also had some



effect on the  subjects.   In support of the above  findings, Torkelson et  al.



(1958) obtained  similar results  with  2 groups of  4 subjects each exposed  to



900 or 500 ppm  of pure  MC  for 75  to 90  minutes.   Subtle changes  in  perceptive



capability were  observed following exposure  to 350  and  450  ppm of MC by



Gamborale and Haltengren (1973)  and Salvini et  al. (1971).   However,  these







002MC3/A                             5-46                             11-10-S1

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          TABLE 5-15.   NON-LETHAL EFFECTS OF METHYL  CHLOROFORM ON  HUMANS
      Study
    Dose/Species
        Effect
Dornette & Jones, 1960
10,000-26,000 ppm
 6,000-22,500
Siebecker et al.,  1960
Stewart et al., 1961
Torkelson et al. ,  1958
Torkelson et al. , 1958
Stewart, 1968; 1971
Am. Ind. Hyg. Assoc.

Stewart et al., 1969
Stewart et al., 1975
Anesthesia
0-2,650 ppm
  for 15 min
500 ppm
900-1000 ppm

900-1,000 ppm
  for 75 win
1,900 ppm for
  5 min

Results of single
  exposures
500 ppm
  6.5-7 hr/day for
  5 days

500 ppm
Induction of anesthesia
Maintain light anesthesia
Depressed BP during mod.  an-
  esthesia
Tendency to develop ventri-
  cular arrhythmias during
  hypoxia (reversed by oxy-
  genation)

Little change in EEC-similar
  halothane anesthesia

See Table 5-1

No effects
See Table 5-2

Slight eye irritation
  and lightheadedness;
Flanagan and Romberg test
  showed slight loss of
  coordination and equili-
  brium
Equilibrium disturbed,
  positive Romberg test

See Table 5-7
Mild subjective
  complaints; abnormal
  Romberg

See Table 5-3
002MC3/A
         5-47
                    11-10-81

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                            TABLE 5-15.   (continued)
      Study
    Dose/Species
        Effect
Gamborale and Hulten-
  gren, 1973


Salvini et al, 1971
Maroni et al., 1977

Seki et al., 1975

Kramer et al., 1978




Weitbrecht, 1965
Stewart and Andrews,
  1966
Stewart, 1971
Litt & Cohen, 1969
250, 350, 450, 550
  ppm for 4-30 min
  periods

450 ppm for 2-4 hr
  periods
110-990 ppm

4, 25, 28, 53 ppm

11-838 ppm •» x = 115
10-20 ppm,
  hands immersed
  in MC
Change in manual dexterity
  and perceptual tests in
  exp. vs.  control groups

Decreased perceptive
  capability under stress,
  eye irritation

No effects

No effects

No effects, although SGPT
  and albumin were different
  in exposed vs. control
  group.

Transient irritation of
  conjunctiva, upper respira-
  tory tract; burning sensa-
  tion in tongue; teeth  felt
  dull (rhubarb effect)
  feeling of ice-cold fingers
  when placed in  liquid  MC;
  hypertension;
  positive urobilinogen;
  automatic dystrophy;
  circulatory dystrophy;
  psychasthenia

See Table 5-4
 002MC3/A
          5-48
                    11-10-81

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last two observations are  questionable  since the effects of  mentol  used to

mask the odor  of  MC in the first study wre not evaluated, and in the second

study the control  subjects were not matched for all  intervening variables such

as food and drinking habits.

     Although  a precise dose-response gradient  of effects produced  by MC

exposure cannot be  derived,  it is possible to establish a crude relationship

between exposure  and effects  from a compilation of the human data available.

A summary  of the  estimated dose-response relationships for acute effects of

single short-term exposures is presented below:

          >5,000 ppm               Onset of narcosis,  possible life threatening

          2,650-1,900 ppm          Lightheadedness,  irritation of the throat

          1,000 ppm                Disturbance of equilibrium

          500-350 ppm              Slight changes in perception, obvious odor

          100 ppm                  Apparent odor threshold

     The approximate concentrations  of  MC which elicit a particular  adverse

effect have greater uncertainty at the two extremes that is,  life threatening
                                                                           *
levels (>5,000 ppm), and  the lowest observed  adverse  effect  level  (LOAEL)

(500-350 ppm).   This results from the inherent difficulties in obtaining exact

exposure values in  the  cases  of accidental overexposure to MC, and the  diffi-

culty  in the acquisition of either quantitative or qualitative data on  slight

behavioral  changes at the lower exposure levels.   Acute overexposure to MC may

be limited by the low odor threshold in humans of 100 ppm.   Others report that

an obvious  odor is present at 500 ppm.   It also should be noted that there are

no reports  of residual  adverse effects from a single exposure to MC.
*
 LOAEL:  is defined as the lowest exposure level in a study or group of studies
which produces statistically significant increases in frequency or severity of
adverse effects between  the  exposed population and  its appropriate control.
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     A major uncertainty  in  the dose-response relationship estimated for MC
from the compilation of the available human reports is the lack of  information
on the length of exposure in accidental  exposure,  and the short duration (less
than 2 hours)  in  most of the studies in which controlled exposures of human
volunteers were used.   Although inhaled  MC is retained in the  lungs in propor-
tion to the concentration in the air.^the time to  reach pulmonary steady-state
conditions is relatively long (4 hours).   Since the maximum body burden from a
given exposure  to  MC  was probably not reached in  any of these studies, it is
likely that  exposures  for longer times  would result in a greater body burden
and  more  severe effects  at  a  given concentration.  In  the studies  of the
inhalation toxicity of MC, there are  insufficient  data to predict the results
of the increased  body burden of MC obtained  by extending the exposure until
steady-state conditions  are  obtained.  However, from the pharmacokinetic data
presented  in Figure 4-1,  it can be  reasonably concluded  that  the effects
observed  in  the dose-response  table would not shift by  more  than 30%.  From
                                t
the above discussion, it should'be evident that these approximations are crude
estimates and contain several areas of uncertainty.
5.4.1.2.   Effects of Intermittent or Prolonged Exposures
     There is  very limited  information from  either animal  or  human  studies  on
the  effects  of subchronic or chronic  inhalation exposure to MC.  As  discussed
below, the only experimental study  of humans  involved  repeated  exposure to  MC
5 days per week for 3 weeks and is of such a short duration that no informa-
tion can  be  extrapolated from this study  as  to  the long term effects of MC
exposure  (Stewart  et al.,  1976).    Observations  of occupationally  exposed
workers are  limited to  reports  where  the  exposure  estimates are imprecise  and
comparable control  populations were  not studied.   The  paucity of chronic
002MC3/A                             5-50                             11-10-81

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toxicity data adds  additional  uncertainty in deriving human risk assessment
based upon inhalation exposure to MC.
     There are,  however,  three subchronic experimental  animal  studies  with
exposures continuing for approximately 90 days which provide some information
on the effects  of  repeated exposures  to MC.  The  study  of Torkelson et al.
(1958) using rats,  guinea  pigs,  rabbits, and monkeys, and the study of Eben
                                                                            *
and Kimmerle (1974)  using  rats established a no observed effect level (NOEL)
for subchronic  exposure to MC for 7 to  8 hours a day, 5 days per week for  3
months at 500 and  440  ppm, respectively.  Similarly,  a  NOEL of 370 ppm was
obtained by Pendergast et  al.  (1967) in  a variety of species (15 guinea pigs,
3 squirrel monkeys, 3 New Zealand rabbits, and 2 beagle dogs) exposed continu-
ously to MC for 6 weeks.   In this study there were 3 deaths in the low exposure
group (135 ppm), but these were attributed to lung infections.  These three
studies would indicate that subchronic exposure to MC in the range of 370 to
500 ppm produces no gross  signs of toxicity.   However, the report of McNutt et
al. (1975) indicates that  exposure below this range may alter some biochemical
parameters in the liver and brain.
     McNutt et  al.  (1975)  observed alterations in the  rough  endoplasmic reti-
culum, detachment  of polyribosomes, microbodies, and triglyceride droplets  in
the livers of mice exposed to 250 ppm of MC continusously for 24 weeks.  At
1000 ppm, the extent of these  changes  had increased and  some individual hepa-
tocyte necrosis was  observed.   Although  it is unclear whether the early bio-
chemical  and histologic changes  observed at exposures of  250  ppm adversely
affected liver  function, they may  represent the first stages in the sequence
of events which leads  to  the hepatocellular necrosis  observed  at 1000 ppm.
*
 NOEL:   is defined  as  the exposure level at which there are no statistically
significant increases  in  frequency or severity of effects between the exposed
population and its appropriate control.
002MC3/A                             5-51                             11-10-81

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The NOEL of 370 to 500 ppm discussed previously may reflect the insensitivity
of the parameters Investigated in the other studies.
     The only study  involving  multiple experimental  exposures of humans was
that of Stewart et al.  (1975),  in which 20 subjects were exposed to 500 ppm of
MC for 7.5 hours a day, 5 days per week for 3 weeks.   There were no effects on
the clinical chemistry of the blood or urine, nor on  pulmonary function tests.
However, the  subjects  did have an increased number of subjective complaints,
including  sleepiness,  irritation, and headaches.  Again,  these  tests  would
probably not  be  sensitive enough to detect  the  slight  changes in the liver
observed in mice at exposures to 1000 or 250 ppm by McNutt et al. (1975).   The
combined evidence  from the  human and animal studies  would indicate that only
minimal effects  are  produced by multiple exposure to 250  ppm  of MC and that
the pathologic importance of these effects are unknown.
     Reports of occupational exposure have major deficiencies which add to the
uncertainty of  a NOEL derived from  these  studies.   The study by Weitbrecht
(1965), which  reported on the  exposure of  9 women  to  MC at a workroom  concen-
tration of 10 ppm, is unacceptable  for human risk assessment.  The number of
workers surveyed was too  small to obtain an  adequate data  base, and the measure-
ments  of  MC  in  the general workroom  air  were  irrelevant for establishing
individual  expsoure  of workers  operating over open  vats  of  a compound as
volatile as MC.  Also, the workers had their hands immersed in the solvent for
varying  lengths  of time,  and it has  been determined  by  Fukabori  et al.  (1976,
1977)  that significant amounts of MC  can  be absorbed through unbroken skin.
Therefore,  while no reliable estimate of  exposure concentration can be made,
it was probably  considerably higher  than the 10  ppm reported.
     The studies of  Maroni  et  al.  (1977),  Seki et  al. (1975) and Kramer et al.
(1978) reported a NOEL for workers occupationally exposed to MC of between 53

002MC3/A                              5-52                              11-10-81

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and 350 ppm.   The  short duration of exposure,  averaging less  than 1 year,  of
the work population in the report of Kramer et  al.  (1978) precludes the  use of
this study in human risk assessment.  The study of  Seki  et al.  (1975)  provides
information on 196 male workers exposed to MC for at least 5 years at  concentra-
tions of 4, 25, 28, and 53 ppm.  The number of  workers exposed at each concen-
tration was  relatively small,  with only 42 people exposed to  the highest
concentration.  While  no dose  related effects were  observed,  the exposed
groups were  only compared among  themselves and not with  a control population.
The reported  details of the analytical procedures were  insufficient to deter-
mine whether the analytical  data represented measurements over the entire time
period (5  years)  of  exposure  of the study population, or whether the  concen-
trations reported  were  the concentrations  in the breathing zone.  Most of  the
deficiences  in  this  study would probably result in an underestimation of the
exposure  levels,  hence, it is  reasonable  to assume that 53 ppm  is a NOEL
concentration  for  MC.   Maroni  et  al.  (1977)  looked  for  neurophysiological
abnormalities  in a population ef 21 women who  had  been exposed for 6.5  years
to an  average concentration  of MC between 110  and  345 ppm.   No neurotoxicity
was observed  in exposed workers, as compared  to a  control  population of  7
unexposed  women.   The  small  size of the exposed and control populations, and
the limited  number of  physiological  measurements made makes the NOEL  obtained
from  this  study extremely uncertain.   However,  the  data of  Maroni et  al.
(1977) do  provide  greater  confidence that long term occupational exposure to
53 ppm of MC produces no adverse effects.
5.4.2  Oral Exposure
     There is  little  information on the  toxicity of MC  by the  oral route.   As
summarized in  Table  5-5,  the  acute  LD5Q  for MC  has been determined in  rats,
mice, guinea pigs, and rabbits by Torkelson et al.  (1958) and the values range

002MC3/A                             5-53                             11-10-81

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from 8,600 to 14,300 mg/kg body weight.   These LDrQ values  could theoretically



be used to calculate an approximate human lethal dose using the cubed root of



the body weight ratios for interspecies conversion (U.S. EPA, 1980; Freireich



et al., 1966; Rail, 1969).  However,  the data obtained from the species  studied



in the report of Torkelson et al.  (1958) do not show a relationship between MC



toxicity and body  weight.   Therefore,  without further  information, it would



not be justified  to place much reliance on  an approximate human lethal dose



calculated using  the above approach.   However, as  a  crude indication of  a



lethal oral  dose  of MC, the  values were calculated from each species with  the



range for a human  lethal dose falling between 770 and 3940 mg of MC/kg.   Also,



there  is  no  information from human exposure to MC to indicate whether values



obtained  from  a  calculated lethal dose are  applicable  to  human toxicology.



For the above  reasons  it  is  considered  inappropriate  to attempt to predict an



approximate human  lethal dose from the present data available.



     There are  no subchronic studies on oral  exposure  to  MC,  and  only one



chronic study,  an NCI  bioassay?  In the NCI (1977) bioassay,  rats and mice



were treated by gavage with MC  in corn oil 5 days/week  for 78 weeks at a level



of  750 and 1500 mg/kg body  weight  for  the  low and high dose group rats,  and



2807  and  5615  mg/kg body weight  for the low and high dose group mice.  Early



deaths were  observed in both  rats and mice, with a statistically  significant



dose-related trend observed in male  and female rats,  and  in female mice.   The



control animals  also had poor  survival and  the early mortality  in this group



would  suggest  that  MC contributed  only partly to  the  early deaths of the



treated  animals.    Another difficulty with  this study  is  that  treatment  of



animals by gavage  is not  identical to exposure  through  food or  water  since the



daily  dose of  the  compound  is  administered at  a single  time, resulting  in  high



body  levels  immediately after treatment.   As a result of  the poor survival of







002MC3/A                             5-54                              11-10-81

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the control animals and the  inappropriateness of the route of administration,
the relevance of this study to human risk assessment is questionable.
5.4.3  Dermal  Exposure
     There is insufficient information available for quantitative risk assess-
ment of dermal  exposure  to MC.  The pharmacokinetic data of Fakabori et al.
(1976, 1977)  would  indicate that absorption of  MC  through  intact skin can
contribute significantly  to the  levels  of  MC  in the  body.  The  extent of
dermal absorption was  estimated  to be 5 percent of respiratory absorption in
humans in direct contact with the liquid.  The high vapor pressure of MC would
preclude dermal  contact with this solvent for  sufficient periods of  time  to
allow toxic quantities  to be absorbed, except in some industrial situations.
Even so, the  greater  efficiency  of absorption  by  inhalation would make this
the route of greatest concern.
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002MC5/F                            6-14                               11-10-81

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002MC5/F                            6-15                               11-10-81

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                                         14
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       7. Appendix





THE CARCINOGEN ASSESSMENT GROUP'S



    CARCINOGEN ASSESSMENT OF





       METHYL CHLOROFORM
              7-1

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                                                           EXTERNAL  REVIEW  DRAFT
                                                           January 16,  1981
                       THE CARCINOGEN ASSESSMENT GROUP'S

                             CARCINOGEN  ASSESSMENT

                                      OF

                               METHYL CHLOROFORM
                                                    Roy  E.  Albert, M.D.
                                                    Chairman
PARTICIPATING MEMBERS

Elizabeth L. Anderson, Ph.D.
Larry D. Anderson, Ph.D.
Steven Bayard, Ph.D.  -
Chao W. Chen, Ph.D.
John R. Fowle III. Ph.D.*
Bernard H. Haberman, D.Y.M.,  M.S.
Charalingayya B. Hiremath, Ph.D.
Chang S. Lao, Ph.D.
Robert McGaughy, Ph.D.
Beverly Paigen, Ph.D.
Dharm Y. Singh, D.V.M.,  Ph.D.
Nancy A. Tanchel, B.A.
Todd W. Thorslund, Sc.D.
Peter E. Yoytek, Ph.D.*

*Reproduct1ve Effects Assessment Group


                                     DRAFT
                              DO NOT QUOTE OR CITE

This document has been reviewed and approved by the Chairman and staff of the
Carcinogen Assessment Group,  Office of Health and Environmental  Assessment, U.S.
Environmental Protection Agency.  It has not been formally released by the EPA
and should not at this stage be construed to represent Agency policy.  It is
being circulated for comment on its technical accuracy and policy implication.


                                    7-2

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                                    CONTENTS

   I.  Summary	1
  II.  Introduction 	  2
 III.  Metabolism	2
  IV.  Mutagenicity and Cell  Transformation 	  4
            Cell  Transformation
   V.  Toxicity	14
  VI.  Carcinogenicity	15
            Rats
            Mice
            Carcinogenicity of 1,4-Dioxane
 VII.  Unit Risk Estimate	23
VIII.  References	26
                                      7-3

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                                  I.   SUMMARY

     There 1s no adequate basis for  the evaluation of the carclnogenidty  of
methyl chloroform.  A lifetime animal  bloassay using both rats  and mice,
currently 1n progress under the National Toxicology Program (NTP),  will be
completed In May 1981.  An earlier National  Cancer Institute (NCI)  study was
Inconclusive due to the poor survival  of treated animals.  An Inhalation study
1n rats by the Dow Chemical Company  showed no  evidence of carclnogenidty, but
the doses were given for only half of the lifetime of the animals  and the
highest dose did not appear to be the maximum  tolerated dose.
    Technical grade methyl chloroform has been shown to be weakly  mutagenic and
to transform animal cells 1n vitro using Fisher rat embryo cell  line F1706.
Technical grade methyl chloroform contains about 3% of the stabilizing substance
dioxane which shows evidence of being a carcinogen (Kociba et al.  1974; Argus et
al. 1965, 1973; NCI 1978).
     Therefore, because of the inconclusive results of the NCI  gavage bioassay
in rats and mice, the dosage Inadequacies in the negative Dow inhalation test
and the possible contribution of dioxane to the positive mutagenic and cell
transformation tests, a final judgment on the  carcinogenicity of methyl
chloroform will have to be deferred until the  results of the NTP bioassay, which
has a very low percentage (0.002%) of dioxane, are available.
                                      7-4

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                               II.  INTRODUCTION
    Methyl chloroform (1,1,1-tHchloroethane,  Cl^CCls)  1s used primarily  as
a cleaning or degreaslng agent for metals.   It 1s Increasingly being  used as  a
substitute for chlorinated ethyl enes.  An estimated 630 million pounds were
produced 1n the United States 1n 1976;  of this,  dispersive uses of methyl
chloroform (primarily metal degreaslng  and aerosols) comprise at least 300
million pounds.  Methyl  chloroform escapes Into the environment primarily Into
the air.  It 1s soluble (2 gm 1n 100 ml)  1n water and has a boiling point of  the
74°C.  The National Institute for Occupational Safety and Health (NIOSH)  judges
that 2.9 million workers may be exposed (Parker et al.  1979).

                                III.  METABOLISM

    Only a small percentage of methyl chloroform 1s metabolized;  most 1s
excreted unchanged by the lungs.  Using rodent liver In vitro, Van Dyke and
Wineman (1971) showed that dechlorf nation of chloroethanes was carried out by
the nrfcrosomal mixed- function oxldases  and required molecular oxygen.  However,
methyl chloroform was a poor substrate  for this reaction compared to  other
chloroethanes and less than 0.5% of the chlorine was enzymatlcally removed.
Metabolism was not Increased when rats  were pre-exposed to methyl chloroform  for
3 days.
    In humans, Monster et al. (1979) have estimated that 60 to 80% of Inhaled
methyl chloroform was exhaled unchanged from the lungs.   Only a small part of
the retained methyl chloroform was excreted 1n urine as trlchloroethanol  (2%)
and trichloroacetlc acid (1.5%).  The authors  had no explanation  for  the
unaccounted methyl chloroform.
    Hake and coworkers (1960) injected  one female and two male rats (170  to
                                     7-5

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                   IV.  MUTAGENICITY* AND CELL  TRANSFORMATION

    Methyl chloroform has been tested for Its ability to cause  point  mutations
in bacteria., point mutations and gene conversion in yeast,  and  for cytogenetic
abnormalities in rats.  The studies presently available for review are
summarized in Table 1.
    Five reports have been prepared about the mutagenicity  of methyl  chloroform
in bacteria; all were conducted using the Salmonella/S9 system.  Two  of  the
reports, Henschler et al. (1977) and Taylor (1977) were reported to be negative.
The report by Henschler cannot be evaluated because no data are presented.   In
the report by Taylor, actually an interoffice memo describing results generated
from testing performed by a contract laboratory, 0, 50, 100, 200,  300, and 500
ul/plate doses of methyl chloroform (source and purity not  reported)  were
administered in plate incorporation tests using TA 1535, TA 1537,  TA  98, and TA
100.  Different concentrations of S9 activation were  also tested.   The high
doses of methyl chloroform (300 and 500 ul/plate) were reported to be toxic  for
all strains in tests conducted without activation.  The 500 ul/plate  dose
level was reported to be toxic for either TA 1537 or  TA 98  1n the  various tests
conducted with metabolic activation, but not for the  other  strains.  The
criteria for determining toxicity were not reported.   Although  special
precautions were not reported to have been taken to prevent evaporation  of
methyl chloroform, it appears that the test agent did enter the cells at the
highest doses because of the toxicity reported.
    ^Prepared by the Reproductive Effects Assessment Group.
                                      7-6

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183 g) Intraperftoneally (1.p.) with 700 rag/kg of 14C-methyl  chloroform.
About 50% of the urinary radioactivity occurred as 2,2,2-trichloroethanol  1n  the
form of glucuronlde conjugate.  The remaining urinary radioactivity  volatilized
at room temperature and was probably the parent compound.   It was  suggested that
methyl chloroform metabolized by an Initial  oxidation to trlchloroethanol  and
subsequent oxidation to small quantities of trlchloroacetlc add.   In  this
study, 98.7% of the Injected radioactivity was exhaled unchanged,  and  0.5% as
14C02.  Only 0.85% of the Injected radioactivity was recovered 1n  urine,  and
only half of that was Identified as a metabolite (Figure 1).

        METHYL
       CHLOROFORM	» TRICHLOROETHANOL
                          oxidation
       CC13 - CH3                             CCla - CH2OH
                                    glucuronlde        \
                                    conjugation          oxidation
                                    /                  \
                    CCla - CH20 - glu.                TRICHLOROACETIC  ACID
                                                             CClsCOOH

          Figure 1.  Metabolic route suggested for methyl  chloroform.
                     Source:  Hake et al. 1960, Ikeda an Ohtsuji 1972

    Ikeda and Ohtsuji (1972) compared the quantities of metabolites  In urine
(trlchloroethanol and trlchloroacetlc add)  1n Wlstar rats exposed by  Inhalation
and by 1ntraper1toneal Injection.  The quantity of metabolites was essentially
equivalent for exposure to 200 ppm for 8 hrs and 2.78 mmol/kg body weight.
These metabolites are the same metabolites formed from trlchloroethylene.
However, the concentration of trlchloroethanol 1n the blood of volunteers
exposed to methyl chloroform 1s only l/35th that found In volunteers exposed  to
equivalent amounts of trlchloroethylene (Monster 1979, Monster et  al.  1979,
Monster and Houtkooper 1979).
                                      7-7

-------
                                            TABLE 1.   MJTAGENICITY TESTING OF METHYL CHLOROFORM
A.  BACTERIA
Reference
Litton 1975

















Henschler
et al. 1977



Simmon et al .
1977








Test System Strain
Salmonella/59: TA 1535
spot test and TA 1537
plate Incor- TA 1538
poration














Sa1mone11a/S9 TA 100




Sa1monella/S9: TA 1535
conducted In TA 1537
sealed TA 1538
desiccator TA 98
TA 100





Activation
System
PCB induced liver,
lung and testes S9
mix from adult male
animals: 1CR random
bred mice, Sprague-
Dawley rats and
Macaca mulatta monkeys,











PCB Induced rat liver
microsome S9 mix.



PCB Induced rat liver
microsome S9 mix.








Concentration
Formulation %
99+ TUT
2.5

96 2.0
1.0

95.65 1.0
0.5

93.75 0.5
0.25






Not reported.




(Extrapolated
from figure 21)
0, 100, 200, 300
400, 500, 750,
1000 ul /9-liter
desiccator.




Result
99+% formulation
reported positive
with TA 1535. Result
was reported to be
repeatable.
The other formulations
were reported to be
negative.










Reported negative.




TA 100
Dose +S9 -S9
, 0" T2T T2T
100 130 145
200 125 180
300 175 185
400 145 -
500 200 220
750 260 210
1000 280 225
Comment
1. All formulations reported
to be not soluble at test
concentrations.
2. Toxicity reported to be
variable (In many tests, 25/48,
as many or more survivors were
observed at the high dose
compared to the low dose).
3. Strains TA 98 and TA 100 not
tested (not available at time
test conducted).
4. No Information about the
Identity of the stabilizers
and other components.
5. No special precaution taken
to prevent evaporation of the
compound, but test reported to
be conducted in suspension.
1. No precautions taken to
prevent evaporation reported.
2. No data presented,
therefore, cannot evaluate.
3. Only one strain tested.
1. Results indicate ability of
methyl chloroform to mutate
bacteria when precautions are
taken to ensure exposure of test
organisms.
2. Data were presented for TA
100 only.
3. Exact purity of test compound
not given, but reported to be
high.
                                                                   7-8

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A.  BACTERIA
                                                          TABLE  1.  (continued)
Reference
Taylor 1977
Test System
Salmonella/59:
plate incor-
poration tests
Strain
TA 1535
TA 1537
TA 98
TA 100
Activation
System
Aroclor-actlvated
rat liver mlcrosome
S9 mix. Different
amounts tested (0,
10, 20, 30, 40, and
50 ul/plate).
Concentration
0, 50, 100, 200,
300, and 500
ul/plate
Result Comment
Reported negative. 1. Mo precautions reported to have
been taken to prevent evaporation
of the test material.
2. Protocol not adequately
described.
3. Mo positive controls reported to
have been run.
4. The purity of the test chemical
not reported.
Snow et al.
1979
Sa1njone]la/S9:   TA 100
conducted 1n
sealed chambers
Methyl  chloroform
Induced Syrian
golden  hamster
liver
S9 mix.
                                                                 0*
                                                               500
                                                               750
                                                              1000
                                                              1500
                                                                 0
                                                               500
                                                               750
                                                              1000
                                                              1500
AldHch Sample   PPG Sample
   (without activation)
144,115,130     144,115,150
234,251,235
271,271,274
                                                           315,270.310
                                                           355,336,340
225,232,233
263,301,262
307,283,268
398,360,336
                                                                 (with activation)
                                                           122,143,119     122,143,119
                                                           324,281,312     266.291.297
                                                                           359,333,348
                                                                           373.372,407
                                  365,346,351
                                  384,402,393
                                  485,432,430
                                                                           471,467,430
                                                                          Linear Regression Analysis
                                                                             (y « ax +b)
1.  Only one strain tested.
2.  Results Indicate ability of
methyl chloroform to mutate bacteria
when precautions are taken to ensure
exposure of test organisms.
3.  Linear dose responses obtained
with no significant differences found
between Aldrlch or PPG samples at any
concentration tested.
4.  Protocol not completely
described.
5.  Purity of Aldrlch sample not
reported.  Purity of the PPG
sample reported to be high but
Information about composition
not provided.
                                                                          Chemical
                                                                          Source
                                                                         B
                                                                          Aldrlch +S9   ^1  I6F .94
                                                                                  -S9   .14  152  .96
                                                                          PPG     +S9   .22  157  .97
                                                                                  -S9   .15  143  .97
          .6-liter B11lups-Rothenberg Modular Incubator Chamber
                                                                                             (continued on the following page)
                                                                  7-9

-------
B.  YEAST
                                                             TABLE  1.   (continued)
Reference
Test System
Strain
Activation
  System
Concentration
                                                                         Result
Comment
Litton 1975   Saccharomyces    D4
              cerevlslae:
              gene conversion
                          PCB induced Sprague-
                          Dawley rat  liver S9
                          mix.
Loprleno
et al. 1979
Schlzosac-
charomyces
pombe forward
mutation
              ade 6-60/   Host mediated assay
              rad 10-198/ B6C3F1 mice.
              h-
                                 Formylatlon   %
                                     59?      T7T~
                                              2.25
                                                                    96
                                                                    95.65
                                                                    93.75
                                                              2.0
                                                              1.0

                                                              2.0
                                                              1.0

                                                              5.0
                                                              2.5
                                   0 or 5000 mg/kg
                                   administered by
                                   gavage to mice.
                                   Yeast exposed for
                                   3, 6, or 16 h.
                                          Reported negative.
                                          Reported negative.
                                         1.   All  formulations  reported to
                                         be  not soluble  at  the test concen-
                                         trations employed.
                                         2.   Toxlclty  reported to be
                                         variable (In  many  cases as many or
                                         more survivors  observed at the high
                                         dose as  at the  low  dose).
                                         3.   No Information  about
                                         stabilizers or  other  chemical
                                         substances In the  formulations.
                                         4.   No special  precautions taken to
                                         prevent  evaporation of methyl
                                         chloroform but  tests  reported to be
                                         conducted in  suspension.

                                         1.   Preliminary results; protocol
                                         not fully presented.
                                         2.   MTO  may not have  been employed.
                                         3.   Unclear if  negative and
                                         positive controls performed
                                         concurrently.
                                         4.   Unclear 1f  test compound was
                                         absorbed from GI tract and exposed
                                         test organisms.
                                         5.   No jm vitro testing performed.
C.MAMMALIAN IN VIVO UTTOGENETICS
Quast         Chromosome     Sprague-
et al. 1978   aberrations    Dawley,
              In rat marrow  spartan
              cells          substrain
                                                   0, 875, and 1750   Reported negative.
                                                   ppm Inhalation
                                                   5 days/week 6 hr/day
                                                   for one year.
                                                                           1.   Protocol  and  data not
                                                                           presented.  Cannot evaluate
                                                                           conclusions.
                                                                           2.   Reported  that data  from female
                                                                           rats Insufficient for clear inter-
                                                                           pretation of  the  results.
                                                                           3.   Treatment had no effect on body
                                                                           weight  of rats.   MTD may not have
                                                                           been employed.
                                                                    7-10

-------
    No special  precautions were reported to have been  taken  to  prevent
evaporation of methyl  chloroform 1n the tests conduced by  L1tton-B1onet1cs
(1975) for Dow Chemical  Company either.  Various formulations of methyl
chloroform (99+%, 96%, 95.65%,  and 93.75%)  were assayed for  mutagenlc potential
1n Salmonella strains TA 1535,  TA 1537, and TA 1538, both  with  and without
metabolic activation.   No Information was provided concerning the stabilizers or
other components of the formulated samples  tested.  Preliminary toxldty  testing
was conducted to determine the  appropriate  test doses.  The  test doses decided
upon differed for each formulation and are  presented 1n Table 1. It was
reported by the contractor that the chemicals tested were  not soluble 1n  the
aqueous testing environment, and 1t was stated, for the 99+% formulation  at
least, that "the toxldty from  test to test was quite  variable  depending  upon
the ability to effectively disperse the compound 1n the testing medium."  The
low solubility of methyl chloroform coupled with Its high  volatility raise the
concern that, In the tests conducted by L1tton-B1onet1cs,  exposure of the test
organisms to 1t may have been minimal.  Salmonella strain  TA 1535 was reported
to exhibit a repeatable mutagenlc response  to the 99+% formulation, but 1t 1s
stated 1n the report that "the  mutagenlclty of this chemical for Indicator
strain TA 1535 must be considered presumptive for the  following reasons:
    A.  The positive response 1s only evident at high  dose levels which
        generally result 1n low population  survivals (high toxlclty).  Thus
        one cannot exclude some type of selection.
    B.  The data from activation plate tests does (sic) not  Indicate any
        activity.  The activation plate tests were repeated  and were negative
        1n those tests as well."
    With respect to point A, an examination of the computer  print-outs
summarizing the mutagenlclty of methyl chloroform for  strain TA 1535 both with
and without activation reveals  that the population of  cells  1n  the high dose
group was equal to or greater than the cells 1n the low dose group 1n two out of
                                      7-11

-------
four cases, Including the experiment in which the highest mutation  frequency  was
observed.  In other words, there does not seem to be  significant  cytotoxicity at
the high dose level which could possibly result in a  spurious  positive  result.
    With respect to point B,  the plate activation tests  performed in  this
experiment were spot tests.  The proper conduct of a  bacterial  spot test
requires that the compound be soluble in an aqueous environment,  dissolving
through the agar from point of placement outward, forming a concentration
gradient.  Since methyl chloroform 1s volatile and was not soluble  in the
testing performed by Lltton-Bionetics, the negative results of the  plate tests
cannot be considered evidence supportive of the nonmutagenicity of  methyl
chloroform: therefore, the study by Litton-Bionetics  (1975) 1s considered  to  be
an inadequate assessment of the mutagenicity of methyl chloroform.
    Two tests of the mutagenic potential of methyl chloroform  in  bacteria  were
reported to have been conducted using protocols designed to prevent evaporation
of methyl chloroform and thereby ensure exposure of the  indicator organisms.
Both tests were reported to yield positive results (Simmon et  al. 1977  and Snow
et al. 1979).  The testing performed by Simmon and coworkers was  conducted using
the standard battery of Salmonella typhlmurium strains TA 1535, TA  1537, TA
1538, TA 98, and TA 100, both with and without PCB-induced rat liver  mlcrosome
S9 mix for metabolic activation.  The concentrations  used for  testing
(extrapolated from figure 21) were 0, 100, 200, 300,  400, 500, 750, and 1000
ug/9-Hter desiccator.  A weak dose-related response was observed for TA 100
both with and without metabolic activation.  These results indicate that methyl
chloroform does possess mutagenic activity 1n Salmonella.  The exact purity of
the methyl chloroform  sample tested was not given but was reported to be high.
    In their studies,  Snow et al. (1979) tested two samples of methyl chloroform
1n Salmonella strain TA 100 both with and without metabolic activation.  Testing
                                      7-12

-------
conducted with metabolic activation employed an S9 mix obtained from methyl
chloroform Induced Syrian golden hamster liver microsomes.   Similar to  the  study
performed by Simmon et al. (1977), precautions were reported to have been taken
to prevent evaporation of methyl chloroform.  Doses of 0, 500,  750, 1000, and
1500 ul/5.6 liter Billups-Rothenberg Modular Incubator Chamber  were employed,
and repeatable, nearly Identical, linear dose-responses were observed for TA 100
(see result column, Table 1) to the two samples of methyl chloroform tested.
One of the samples was from Aldrich (97% methyl chloroform  stabilized with  3%
p-d1oxane) and the other was from PPG Industries (reported  to be a purified
sample).  The results of Snow and coworkers strengthen the  observations by
Simmon et al. (1977) that methyl chloroform 1s a weak mutagen 1n Salmonella.
The nearly Identical responses achieved after testing these two samples of
methyl chloroform obtained from different sources support the conclusion that
methyl chloroform Is a mutagen.
    Testing of methyl chloroform for mutagenldty employing yeast as Indicator
organisms has been conducted by two laboratories (L1tton-B1onet1cs 1975 and
Loprleno et al. 1979).  Both reported negative results. The L1tton-B1onet1cs
yeast study was conducted at the same time as was the testing of methyl
chloroform In bacteria, and many of the deficiencies noted  above concerning the
bacterial tests apply to the yeast testing as well.  No special  precautions were
reported to have been taken to prevent evaporation of the test  compound;  the
compound was not soluble under the condition of test, and the toxlclty  results
were reported to be variable.  In many cases (I.e., 5/11 for the various
formulations) as many or more survivors were observed at the high dose  as were
observed at the low dose.  As was the case for the bacterial results, the
testing by Lltton-Blonetlcs of methyl  chloroform with respect to Its ability to
Induce gene conversion 1n Saccharomyces cerevlslae with and without rat liver S9
                                      7-13

-------
mix metabolic activation is considered to have resulted in a "no-test."   The
volatility and insolubility of methyl  chloroform raise the likelihood that the
cells on test were simply not adequately exposed.
    Based on preliminary testing using the host-mediated assay  in  B6C3F1  hybrid
mice, Loprieno et al. (1979) have reported that methyl chloroform  administered
by gavage at 500 mg/kg did not increase the incidence of forward mutations in
Schizosaccharomyces pombe measured after treatment times of 3,  6,and 16 hours.
No Information was provided concerning the ability of methyl  chloroform to be
absorbed and transported to the peritoneum, thereby effectively exposing  the
yeast cells, and no information is provided concerning whether  or  not testing
was conducted to determine the ability of methyl chloroform to  induce mutations
in vitro.  Furthermore, no data are presented concerning a determination  of the
toxicity of the substance to mice after acute exposure to arrive at a maximum
tolerated dose for conducting the host-mediated assay.  Besides the concerns
that the Indicator organisms may not have been exposed to methyl chloroform, the
report does not provide adequate information concerning the design and conduct
of the testing and this In turn, makes it Impossible to assess  the significance
of the results.  (It is not clear what protocol was followed to plate the yeast
after harvest; it Is not clear if concurrent positive and negative controls were
performed, etc.)
    The reported lack of chromosome aberrations produced after  chronic exposure
to Sprague-Dawley rats (Quast et al. 1978) also cannot be evaluated based on the
data provided to EPA at the time of writing this report.  Details  concerning the
protocol employed are very sketchy.  The conclusions are forwarded unsupported
by the presentation of experimental data.*
     *A written request has been made to Dow to obtain these data.
                                       7-14

-------
    A joint study sponsored by Imperial Chemical  Industries and the Medical
Research Council in the United Kingdom, and the National  Institute of
Environmental Health Sciences in the U.S.A. entitled the  International  Program
for Evaluating Short-Term Tests for Carcinogenicity (IPESTTC)  was recently
conducted to assess the ability of short-term tests (including mutagenicity
tests) to predict carcinogenicity.  Forty-two chemical  substances of high purity
were tested blind in 23 different assays, (e.g.,  gene mutation tests in
bacteria, yeast, and mammalian cells in culture;  SCE formation in vitro and  jm
vivo; chromosome aberrations 1n -vitro, etc.) some of which were conducted in
more than one laboratory.  The results of the IPESTTC testing  have recently  been
published (8/6/81).  EPA has not yet received a copy of this report to  evaluate
the data, but preliminary indications are that methyl  chloroform tested negative
in a majority of the experiments (apparently only 9 weak  positive responses  were
obtained out of 38 test systems).  When EPA receives these data, they will be
evaluated with respect to the mutagenicity of methyl cloroform.  Since
preliminary reports of the IPESTTC study do not note which, if any, precautions
were taken to ensure exposure of the test organisms to the compound, and the
data are not yet releasable for complete review,  the preliminary indication  that
methyl chloroform tested negative in the IPESTTC study needs to be further
evaluated.
    In summary, methyl chloroform has been tested for its mutagenic potential in
several systems (bacteria, yeast, and rats), but  sufficient data are not
presented for evaluating the results presented in three papers (the Dow 1978 rat
cytogenetic study, the Henschler et al. 1977 testing in Salmonella, and the
Loprieno et al. 1980 testing in yeast).  Of the remaining papers, only  two
described the results of mutagenicity testing conducted in such a manner as  to
prevent evaporation of methyl chloroform and to ensure exposure of the  indicator
organisms.  Both of these tests (Simmon et al. 1977 and Snow et al. 1979) were
conducted in Salmonella and a weak positive response was  reported for both.
                                      7-15

-------
     On the basis of the available evidence,  it  is  concluded  that methyl
chloroform is mutagenlc in bacteria.   If the  metabolism  and pharmacokinetics of
this compound in humans results in metabolic  products which can interact with
DNA as is the case for bacteria, it may  cause mutagenic  effects in humans as
well.  However, additional testing in other organisms, (e.g., mammalian cells  in
culture) is necessary to confirm the  mutagenicity observed in bacteria.  Careful
attention should be given to the design  and conduct of these  studies  to prevent
the evaporation of methyl chloroform, overcome its  low solubility in  aqueous
media, and to ensure exposure of the  indicator organisms.

CELL TRANSFORMATION
    Price et al. (1978) exposed Fischer  rat embryo  cell  cultures  (F1706,
subculture 108) to 1,1,1-trichloroethane liquid  at  concentrations of  9.9 x  IQl
and 9.9 x 102 uM for 48 hours.  1,1,1-Trichloroethane was diluted with growth
medium to yield the appropriate doses.  The 1,1,1-trichloroethane sample
obtained from the Fisher Scientific Company was  purportedly >_ 99.94 pure, but  a
personal communication from Carlson of the Fisher Scientific  Company  revealed
that methyl chloroform supplied by them  to Dr. Price was really the product of
the Dow Chemical Company which is about 95% pure.   The chemical composition of
Dow's methyl chloroform is given in Table 4,  as  reported by Quast et  al.  (1978)
of the Dow Chemical Company.  The cells  were  grown  in  Eagles  minimum  essential
medium In Earle's salts supplemented with 10% fetal bovine  serum, 2 mM
L-glutamine, 0.1 mM nonessential amino adds, 100 ug pencillin, and 100 ug
streptomycin per ml.  Quadruplicate cultures  were  treated at  50%  confluency with
each dose.  After treatment, cells were cultured in growth  medium alone at  37°C.
Transformation of cells treated with either dose level  of 1,1,1-trichloroethane
was observed by 23 and 44 days of incubation  and was characterized  by
                                       7-16

-------
progressively growing foci composed of cells lacking contact Inhibition  and
orientation.  There was no transformation of cells grown in medium alone or in
the presence of a 1:1000 acetone concentration.  Fifty-two and 55 microscopic
foci per three dishes with low and high 1,1,1-trichloroethane dose,
respectively, were found in dishes inoculated with 50,000 cells from cultures
treated four subcultures earlier and held for four weeks at 37°C in a humidified
C02 incubator prior to staining.
      Subcutaneous injection of cells treated with 9.9 x 101 uM
1,1,1-trichloroethane two subcultures earlier produced local flbrosarcomas in
8/8 newborn Fischer 344 rats within 68 days following treatment.  The ability of
cells grown in growth medium alone to induce local fibrosarcomas was not
determined; however, this tumor type did not develop in rats given cells grown
in the presence of a 1:1000 concentration of acetone.  Exposure of cells to 3.7
x 10~1 uM 3-methylcholanthrene produced 124 microscopic foci per three dishes
in the inoculation test described above by 37 days of incubation and local
fibrosarcoma in 12/12 rats by 27 days following subcutaneous injection of cells.
The exposure of 3-methylcholanethrene was attained by initial  dilution 1n
acetone to 1 mg/ml followed by further dilution in growth medium to 0.1  ug/ml
(personal communication, Dr. Price).  It is understood that this cell  line
contains the genome of the Rausher leukemia virus, but there 1s no basis for
minimizing the positive results since the mode of action of
1,1,1-trichloroethane is not known i.e., due to activation of the virus.

                                  V.  TOXICITY

    The toxicity of methyl chloroform has been recently reviewed (Parker et al.
1979).  Experiments were performed with various species and strains of animals
at different concentrations.  The most harmful  effects of methyl  chloroform are
                                       7-17

-------
central nervous system problems including anesthesia,  disturbed  equilibrium,  and
impairment of perceptual  speed and dexterity.   The effects  of methyl  chloroform
on the heart include bradycardia,  hypotension,  and cardiac  arrhythmias.
Exposure can also cause inflammatory changes in the lungs,  cellular damage  and
fatty changes in the liver, and damage to the kidneys.  Methyl chloroform is
reportedly teratogenic to chick embryos (Elovaara et al. 1979).

                              VI.   CARCINOGENICITY

 RATS
    Two rat bioassays have been completed and one is in  progress.   The completed
studies include an NCI bioassay and an inhalation study  by  the Dow Chemical
Company.  A 2-year carcinogenesis  bioassay by gavage is  now underway by the NTP
in the mouse and rat with 1,1,1-trichloroethane, which has  a very  low percentage
of dioxane.  The animals will be sacrificed in May 1981.  In the previous NCI
bioassay, technical grade 1,1,1-trichloroethane was used.  This  was purchased
from Aldrich Chemical Company, Inc., Milwaukee, Wisconsin.   The  purity was
checked by Hazleton Laboratories of America, Inc., Vienna,  Virginia using
gas-liquid chromatograph (glc) and infrared spectrophotometry.   Analyses by glc
showed that it contained 95% 1,1,1-trichloroethane and 3% p-dioxane, an
inhibitor routinely added to commercial preparations of 1,1,1-trichloroethane.
The remaining 2% of the glc peak area contained several  minor impurities, two of
which may have been 1,1-dichloroethane and 1,1-dichloroethylene.  In this study,
Osborne-Mendel rats were treated with 750 mg/kg and 1500 mg/kg of  methyl
chloroform in corn oil 5 times a week for 78 weeks by gavage (NCI  1977).  The
rats were observed an additional 32 weeks with the experiment ending at 110
weeks.  Both males and females were used with 20 untreated males and 20
untreated females and 50 of each sex at each dose.  The study was  inadequate
because only 3% of the treated rats survived to the end of the experiment.
                                       7-18

-------
    The survival of both sexes of dosed rats was less than that of the matched
control groups, which was also inadequate in the males.   In male rats 6/20 (30%)
of the controls, 32/50 (64%) of the low-dose group, and  36/50 (72%)  of the
high-dose group died within a year of the start of the study.  The Tarone
statistical test of survival showed a dose-related positive trend (P < 0.001)  in
the proportions of deaths over the period of the experiment, although this
differential mortality is not reflected in the survival  at 78 weeks.  In female
rats, 1/20 (5%) of the matched controls, 24/50 (48%) of  low-dose group, and
21/50 (42%) of the high-dose group died in the first year.  As in male rats,  the
statistical test for positive dose-related trend was significant (P < 0.04).   In
both sexes, the early mortality in the 1,1,1-trichloroethane-treated rats may
have affected the incidence of late-appearing tumors; this is especially true  in
the males, since none survived to the scheduled termination of the study.
    Fewer of the rats receiving 1,1,1-trichloroethane survived at both 78 and
110  weeks than did the positive control rats receiving  the known carcinogen
carbon tetrachloride (see Table 2).
               TABLE 2.  COMPARISON OF SURVIVAL OF CONTROL GROUPS,
        1,1,1-TRICHLOROETHANE-TREATED, AND CARBON TETRACHLORIDE-TREATED
                            (POSITIVE CONTROL) RATS
1 ,1 ,1-Trichl oroethane
Group
Male
control
low dose
high dose
Femal e
control
low dose
high dose
Initial
No. of
Animals

20
50
50

20
50
50
Number
Alive at
78 Weeks

7
1
4

14
9
12
Number
Alive at
110 Weeks3

0
0
0

3
2
1
Carbon Tetrachloride
Initial
No. of
Animal s

20
50
50

20
50
50
Number
Alive at
78 Weeks

20
34
35

18
38
21
Number
Alive at
110 Weeks3

12
15
8

14
20
14
          in study at last weighing.
                                      7-19

-------
A variety of neoplasms (Table 3)  were represented In  1,1,1-trichloroethane-
treated and matched-control  rats.  However,  each  type of  neoplasm  has been
encountered previously as a  lesion 1n untreated rats.  The neoplasms observed
are not believed attributable to  1,1,1-trlchloroethane exposure, since no
relationship was established between the dosage groups, the  species, sex, type
of neoplasm, or the site of  occurrence.   Even  1f  such a relationship were
Inferred, It would be Inappropriate to make  an assessment of the cardnogenlcity
of 1,1,1-trlchloroethane on  the basis of this  test, because  of the abbreviated
Hfe spans of the rats.  The NCI  Clearing House on Environmental Carcinogens
concluded that the cardnogenlcity could not be determined from this study (NCI
Clearing House 1977).
    The Dow Chemical study (Quast et al. preprint 1978) treated groups of
Sprague-Dawley rats by Inhalation under  conditions that were slmlllar to those
experienced by workers (6 hours/day, 5 days/week, over one-half of a lifetime).
Rats were treated 12 months  and observed until death  or until they reached the
age of 31 months.  The dose  of 875 and 1750  ppm was 2.5 and  5 times the
threshold limit value of 350 ppm.  Composition of formulation of
1,1,1-trlchloroethane 1s given In Table  4.  Total tumor Incidence  In the treated
animals was similar to that  of controls  (Table 5).
    When tumors at each site by tumor type,  both  benign and  malignant, were
examined, there were eight differences between control and treated animals at
the P < 0.05 level (Fisher Exact  Probability Test).   Seven of these were
decreased tumor Incidence; one was an Increase In ovarian granulosa cell tumors
1n females at the 875 ppm dose.  There were  no ovarian tumors In 189 controls,
three 1n 33 treated at 875 ppm (P = 0.003),  and  two  In 82 treated  at-1750 ppm  (P
= 0.14).  Since ovarian granulosa cell tumors  are rare and the  P value was low
for the animals treated at 875 ppm, 1t will  be of Interest to see  whether this
result Is repeated 1n the NTP study.
                                       7-20

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    TABLE 3.  STATISTICAL ANALYSES OF THE  INCIDENCE  OF  TUMORS  AT  SPECIFIC  SITES  IN  MATCHED CONTROLS  AND
                                      1,1,1-TRICHLOROETHANE-TREATED  RATS
Topography: Morphology
Total Animals: all tumorsb
P Values0
Weeks to first observed tumor
Pituitary: Chromophobe
Adenomab
P Values0
Weeks to first observed tumor
Thyroid: Follicular-Cell
Adenomab
P Values0
Weeks to first observed tumor
Adrenal: Cortical
Adenomab
P Values0
Weeks to first observed tumor
*»• i i i i i
Matched
Control
3/20
N.S.
72
0/20
N.S.
--

0/20
N.S.
— —

0/20
N.S.

Males3
Low
Dose
6/48
N.S.
28
0/48
N.S.
--

0/48
N.S.
— -

3/49
N.S.
28
High
Dose
6/50
N.S.
50
0/48
N.S.
—

0/50
N.S.
--

1/50
N.S.
106
Matched
Control
7/20
N.S.
58
3/20
N.S.
—

2/20
N.S.
--

2/19
N.S.
85
Females3
Low
Dose
7/50
N.S.
64
2/48
N.S.
—

0/50
N.S.
--

1/48
N.S.
99
High
Dose
9/50
N.S.
56
1/48
N.S.
—

1/49
N.S.
--

2/49
N.S.
106
doses of 750 and 1,500 mg/kg body  weight,  respectively.

    bNumber of tumor-bearing animals/number  of  animals examined at  site.

    °Beneath the incidence of the  matched  controls  is the probability level for the Armitage test for
positive dose-related trend in proportions when it  is below 0.10, otherwise N.S. = not significant.
Beneath the dosed group incidence  is  the probability level for the  Fisher exact (conditional) test for
comparison of that dosed group with the matched control  group when  it is below 0.10, otherwise N.S. = not
significant.
                                                    7-21

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    TABLE 4.   COMPOSITION OF THE FORMULATION OF 1,1,1-TRICHLOROETHANE
               USED IN CHRONIC  INHALATION  STUDIES IN  RATS
                          (Quast et al.  1978)
Compounds3
1 , 1 , l-Tr1 chl oroethane
Nltrome thane
Butyl ene oxide
l,4-D1oxane
Liquid volume %
94.71
0.44
0.74
3.93
Calculated weight %
95.88
0.38
0.46
3.11
••Analysis of 1,1,1-trfchloroetnane Lot TA020T3B oy gas cnromatograpny.
    TABLE 5.  TUMOR INCIDENCE IN RATS TREATED WITH METHYL CHLOROFORM
                          (Quast et al. 1978)


control
875 ppm
1750 ppm
Number
male
189
91
93
of Animals
female
189
92
93
Total
male
1.06
0.85
1.11
Neopl asms/Animal
female
2.97
2.67
3.23
                                  7-22

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    The Dow study suffers from two drawbacks:   1)  the animals  were treated for
only 12 months rather than a lifetime but they were observed for another  12
months, and it is not evident that the maximum tolerated dose  was used during
the treatment period.  There is no evidence that a range-finding study
(subchronic) has been done before the start of experiment.   The treated animals
in the Dow study were no different in body weight, terminal  organ weight,  or
mortality from untreated animals.  The only sign of toxicity was an increased
incidence of focal hepatocellular alterations in female rats at the highest
dosage.

MICE
    In the NCI bioassay (NCI 1977), B6C3F1 hybrids were used with 20 animals of
each sex in the control group and 50 animals of each sex at each treatment dose.
The time-weighted average dose was 2807 mg/kg and 5615 mg/kg.   The mice were
treated by gavage 5 days a week for 78 weeks and observed for another 12  weeks
for a  total of 90 weeks in the experiment.
    In male mice, 10/20 (50%) of the matched-control group, 21/50 (42%) of the
low-dose group, and 25/50 (50%) of the high-dose group died within a year of the
start  of the experiment.  In female mice, 1/20 (5%) of the matched-control
group, 9/50 (18%) of the low-dose group, and 20/50 (40%) of the high-dose group
died within the first year of the study.  The Tarone test for positive
dose-related trend in the proportions surviving had a significance level  of
P  = 0.002, although this differential mortality is not reflected in the survival
at 78  weeks.  Table 6 shows that a few mice receiving carbon tetrachlorfde
survived until the planned termination of the test, and 25 to 40% of those
treated with 1,1,1-trichloroethane reached the planned termination date.   The
                                        7-23

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              TABLE 6.   COM>ARISON  OF SURVIVAL  OF  CONTROL  GROUPS,
        1,1,1-TRICHLOROETHANE-TREATED, AND  CARBON  TETRACHLORIDE-TREATED
                            (POSITIVE CONTROL)  MICE
1 ,1 ,1-THchl oroethane


Group
Male
control
low dose
high dose
Femal e
control
low dose
high dose
Initial
No. of
Animals

20
50
50

20
50
50
Number
Alive at
78 Weeks

6
21
14

12
28
14
Number
Alive at
90 Weeks

2
15
11

11
23
13
Carbon Tetrachlorlde
Initial
No. of
Animals

20
50
50

20
50
50
Number
Alive at
78 Weeks

13
11
2

18
10
3
Number
Alive at
90 Weeks

7
0
1

17
0
1
high early mortality In mice receiving 1,1,1-trlchloroethane may  have  lowered
the Incidence of late-appearing tumors.  The treated animals gained  less  than
the controls.
    A variety of neoplasms (Table 7)  were represented 1n 1,1,1-trichloroethane-
treated and matched-control  mice.  However,  each type of neoplasm has  been
encountered previously as a  lesion In untreated mice.  In male  mice  there
appeared to be an excess of  tumors In the liver with one occurring among  the
controls and four among the  treated,  but the Increase was not statistically
significant.  The neoplasms  observed  are not believed attributable to
1,1,1-trlchloroethane exposure, since no relationship was established  between
the dosage groups, the species, sex,  type of neoplasm, or the site of
occurrence.  It would be Inappropriate to make an assessment of cardnogenlcity
of 1,1,1-trlchloroethane on  the basis of this test,  because of  the abbreviated
life spans of the mice.
                                       7-24

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    TABLE  7.   STATISTICAL  ANALYSES OF THE INCIDENCE OF  lUMORS AT SPECIFIC SITES IN MATCHED CONTROLS AND
                                     1,1,1-TRICHLOROETHANE-TREATED MICE
Topography : Morphol ogy
Total Animal: All tumors^
P Values0
Weeks to first observed tumor
Hematopoletlc System:
Malignant Lymphoma^
P Values0
Weeks to first observed tumor
Liver: Hepatocellular
Adenoma or Carcinoma
or Neoplastic Nodule"
P Values0
Weeks to first observed tumor
Matched
control
2/15
N.S.
80
2/15
N.S.
80


0/15
P = 0.035

Males3
Low
Dose
2/47
N.S
89
0/47
N.S.
— —


0/47
N.S.

\
High
Dose
6/49
N.S.
50
2/49
N.S.
64


4/49
N.S.

Matched
Control
4/18
N.S.
80
3/18
N.S.
80


0/18
N.S.

Females3
Low
Dose
2/48
N.S.
54
1/48
N.S.
90


0/48
N.S.

High
Dose
3/50
N.S.
26
0/50
N.S.
»•*


0/50
N.S.

time-weighted average doses of 2,807  and  5,615 mg/kg/ body weight, respectively.

    ^Number of tumor-bearing animals/number  of animals examined at site.

    cBeneath the matched controls  Incidence  1s the  probability level for the Armltage test for positive
dose-related trend 1n proportions  when  1t Is below  0.10, otherwise N.S. = not significant.
Beneath the dosed group Incidence  1s  the  probability level for the Fisher exact test for comparison of
that dosed group with the control  group when It  Is  below 0.10, otherwise N.S. = not significant.
                                                      7-25

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CARCINOGENICITY OF 1,4-DIOXANE
    Methyl chloroform contains a small  amount of stabilizing substances.   The
concentration of specific stabilizers that had been identified in various
commercial methyl  chloroform products is shown below (Avlado 1977 and Detrex
1976, cited by Mazaleskl 1979).
                                           Volume %
       Nitromethane                        0.4 - 1.8
       Butylene oxide                      0.4 - 0.8
       Dioxane                             2.5 - 3.5
       Dloxolane                           1.0 - 1.4
       Methyl ethyl ketone                 1.0 - 1.4
       Toluene                             1.0 - 1.4
       2-Butyl alcohol                     0.2 - 0.3
       Isobutyl alcohol                     1.0-1.4
Not all of these stabilizers are 1n every product, but the maximum total
Inhibitor package (combinations of stabilizers) appears to be between 7 and 8%
by volume (Aviado 1977,  cited by Mazaleski 1979).
    Since dioxane Is a contaminant in methyl  chloroform (about 3%) the
cardnogenlcity of dioxane has been studied extensively.  The results of these
studies are summarized Table 8.
    It should be noted that dioxane causes liver and nasal  tumors in more  than
one strain of rats and hepatocellular carcinomas 1n mice.   Liver tumors have
been Induced by dioxane In both male and female rats as well as In mice.   These
animal results, coupled with a reported finding of nasal carcinomas in furniture
workers exposed to dioxane (NCI 1978, p. 108), suggest that dioxane 1s a
potential human carcinogen.  A detailed evaluation of the  cardnogenicity  of
1,4-dloxane is currently being prepared by the CAG.

                            VII.  UNIT RISK ESTIMATE

    No data exist which can be used to estimate quantitatively the potential
human carcinogenicity of methyl chloroform.
                                      7-26

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TABLE 8.   SUMMARY OF ANIMAL CARCINOGENICITY STUDIES FOR DIOXANE
Species
Rats



Rats



Guinea
Pigs
Rats



Mice



Rats






Strain
Wlstar



Sprague-
Dawley de-
rived Charles
River CD


Sprague-
Dawley de-
rived Charles
River CD
Swiss Webster



Sherman






Route, Frequency of
Administration Sex Control
In drinking water M Untreated
for 63 weeks


In drinking water M Untreated
for 13 months


In drinking water M Untreated
for 23 months
In drinking water M Untreated
for 13 months


Applied to shaved M Acetone
skin of back 3 F Acetone
times/week for 60
weeks
In drinking water M Untreated
for 2 years 4
F




Dose
300 Rig/day
average


0.75, 1.0.
1.40, and
1.8%

588-635 g
In 23 mo.
0.75, 1.0,
1.40, and
1.80%

Unspecified
Unspecified


0.01, 0.1
and It





Tumor type
Hepatocellular carcinoma,
renal transitional cell
carcinoma, my el old leukemia
and lymphosarcoma
Hepatocellular carcinoma
(dose-response) 4/30, 9/30,
16, 30, 23/30

Gall bladder carcinoma.
hepatoma, renal adenoma
Squamous cell carcinoma (nasal
cavity), heptacellular
carcinoma and fibroma

Carcinoma, subcutaneous tumor



Hepatic tumors P * 00022
(all types)
Hepatocellular P - 00033
carcinoma
Nasal carcinoma P * 054
(these tumors are at
the U level)
Reference

Argus et al.
1965

Argus et al .
1973


Hoch-Llgetl
and Argus 1970
Hoch-Llgetl
et al. 1970


King et al.
1973


Koclba et al.
1974





                                                                 (continued on  following page)
                              7-27

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                                                       TABLE B.  (continued)
                      Route, Frequency of
Species Strafn
Rats Wlstar
Administration
Inhalation 7 hrs
dally 5 day /week
Sex
M
&
F
Control Dose
Filtered 111 ppm
air
Tumor type
No hepatic or nasal carcinoma
Retlculum cell sarcoma
M 18/150 vs. 37/206 (P < 0.08)
F 18/139 vs. 30/207
Found many other types of tumor
but not significant
Reference
Torkelson et
al. 1974
Rats
Mice
Osborne-
Mendel
B6C3F1
In drinking water
for 110 weeks
In drinking water
for 90 weeks
M
t
F
N
«
F
Untreated      0.5 and         Hepatocellular adenoma (P • 0.001)
               l.OJ            and squamous cell carcinoma
                               of  the  nasal turblnataes
                               (P  - 0.008) (both sexes)

Untreated      0.5 and         Hepatocellular carcinoma
               l.OS            (P  * 0.001) (both sexes)
NCI Bloassay
1978
                                                               7-28

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                                VIII.   REFERENCES
Argus, M.J., J.C. Arcos, and C.  Hoch-L1geti.   1965.   Studies  on  the
   carcinogenic activity of protein-denaturing agents:  Hepatocarcinogenicity
   of dioxane.  J. Nat!. Cancer Inst.   35:949-958.

Argus, M.F., R.S. Sohal, G.M. Bryant,  C.  Hoch-Ligeti,  and J.C. Arcos.   1973.
    Dose-response and ultrastructural  alterations in  dioxane  carcinogenesis.
    Eur. J. Cancer 9:237-243.

Elovaara, E., K. Hemminki, and H. Vainio.  1979.   Effects of  methylene
    chloride, trichloroethane, trichloroethylene, tetrachloroethylene,  and
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Hake, C.L., T.B. Waggoner, D.N.  Robertson,  and V.K. Rowe.  1960.   Metabolism
    of 1,1,1-trichloroethane by rats.   Arch.  Environ.  Health. 1:1010

Henschler, D., E. Eder,  T. Neudecker,  and M.  Metzler.   1977.  Carcinogenicity  of
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Hoch-Ligeti, C., and M.F. Argus.  1970.  Effect of carcinogens on  the lung  of
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Hoch-Ligeti, C., M.F. Argus, and J.C.  Arcos.   1970.   Induction of  carcinomas
    in the nasal cavity of rats by dioxane.  Brit. J.  Cancer  24:164-170.

Ikeda, M., and H. Ohtsuji.  1972.  Comparative study  of the excretion of
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King, M.E., A.M. Shefner, and R.R. Bates.  1973.   Carcinogenesis bioassay of
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Kociba, R.J., S.B. McCollister,  C. Park,  T.R. Torkelson,  and  P.J.  Gehrfng.
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Litton Blonetics, Inc.  1975.  Mutagenic evaluation of compound  D6.  LBI
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Loprieno, N., R. Banale, A.M. Rossi, S. Fumero, G. Mer1gg1, A. Mondlno, S.
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Mazaleski, S.C.  1979.  An assessment of the  need for limitation on
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    560/11-79-009 p. 3-17.
                                      7-29

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Monster, A.C.  1979.   Difference 1n uptake,  elimination,  and metabolism  1n
    exposure to trichloroethylene,  1,1,1-trichloroethane  and
    tetrachloroethylene.  Int.  Arch. Occup.  Environ.  Health 42:311-317.

Monster, A.C., 6. Boersma, and  H. Steenweg.   1979.  Kinetics of
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Monster, A.C., and J.M. Houtkooper.  1979.   Estimation of Individual  uptake of
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    biological parameters.  Int. Arch. Occup. Environ. Health 42:319-323.

NCI.  1977.  Bioassay of 1,1,1-trichloroethane for  possible cardnogenlcity.
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NCI.  1978.  Bioassay of l,4-d1oxane for possible carcinogenlcity.  Carcinogen
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NCI Clearinghouse on Environmental  Carcinogens.  March 25, 1977.
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Parker, J.C., G.E. Casey, L.J.  Bahlman, N.A. Leidel,  D. Rose,  H.P.  Stein, A.VI.
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Quast, J.F., L.W. Rampy, M.F. Balmer, B.D.J. Leong, and P.J. Gehring.  1979.
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Simmon, V.F., K. Kauhanen, and R.G. Tardlff.  1977.  Mutagenic activity of
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Snow, L.P., B.C. MacNair, B.C. Castro.  1979.  Mutagenesls testing  of
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Taylor, G.  1977.  Mutagen1c1ty  testing: Interoffice memo to Office/Division
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Torkelson, T.R., B.K.J.  Leong, R.J. Kociba, W.A. Rlchter, and P.J.  Gehring.
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U.S.  Environmental Protection Agency  (EPA).  1976.    Interim procedures and
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Van Dyke,  R.A.,  and C.G. Wineman.   1971.  Enzymatic  dechlorlnatlon.
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                                       7-30

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