EPA 560/11-79-006
Investigation  of Selected  Potential
    Environmental  Contaminants:
Ethylene  Glycol, Propylene Glycols
        and  Butylene  Glycols
                      By:
                   Lynne M. Miller
                     May 1979

                    Final Report
                 EPA Contract No. 68-01-3893
                  FRC No. 80G-C4807-01
                    Project Officer:

                    Robert Carton
                    Prepared For:
                Office of Toxic Substances
             U.S. Environmental Protection Agency
                 Washington, D.C. 20460
              Franklin Research Center
              A Division of The Franklin Institute
              The Benjamin FrankSn Partway. Phila.. Pa. 19103 (215) 448-1000

-------
                                          EPA 560/11-79-006
Investigation of Selected  Potential
    Environmental Contaminants:
Ethylene  Glycol, Propylene Glycols
         and  Butylene  Glycols
                        By:
                     Lynne M. Miller
                       May 1979
                      Final Report
                  EPA Contract No. 68-01-3893
                    FRC No. 80G-C4807-01
                  Document is available to the
                    public through the
              National Technical Information Service
                  Springfield, Virginia 22151
                     Project Officer:
                     Robert Carton
                     Prepared For:
                 Office of Toxic Substances
              U.S. Environmental Protection Agency
                  Washington, D.C. 20460
          JuUU Franklin Research Center
              A Division of The Franklin Institute
              The Benjamin Franklin Parkway, Phila.. Pa. 19103 (215) 448-1000

-------
                            TABLE OF CONTENTS


                                                                  Page


PREFACE                                                            xii


LIST OF TABLES                                                     xiii


LIST OF FIGURES                                                    xv


EXECUTIVE SUMMARY                                                  xvi


I.  PHYSICAL AND CHEMICAL DATA                                     1


    A.  Chemical Structure                                         1


    B.  Properties of the Pure Material                            5


    C.  Properties of the Commercial Material                       8


    D.  Chemical Reactions Involved in Use                        10


II. ENVIRONMENTAL EXPOSURE FACTORS                                15


    A.  Production                                                15


        1.  Production Processes                                  15


            a.  Ethylene Glycol                                   15


            b.  Propylene Glycols                                 17


            c.  Butylene Glycols                                  19


        2.  Quantity Produced                                     21


        3.  Domestic Producers and Production Sites               21


        4.  Imports and Foreign Producers                         27


        5.  Market Price                                          31


    B.  Use                                                       31


        1.  Ethylene Glycol                                        31


            a.  Current Demand                                    31


            b.  Market Trends                                      31

                                                                   O Q
        2.  Propylene Glycols
                                   11

-------
                                                             Page




        a.  Current Demand                                    38




        b.  Market Trends                                     39




    3.  Butylene Glycols                                      42




C.  Possible Alternatives to Use                              43




D.  Entry into the Environment                                44




    1.  From Production                                       44




    2.  From End Product Manufacture                          45




    3.  From Use and Disposal                                 45




    4.  From Transportation                                   47




    5.  From Microorganisms                                   48




    6.  From Other Sources                                    50




E.  Waste Handling                                            50




    1.  Characteristics of Waste Stream                       50




    2.  Treatment of Waste Stream                             53




    3.  Diol Recovery                                         55




F.  Fate and Persistence in the Environment                   56




    1.  Biological Degradation                                56




        a.  Ethylene Glycol                                   56




            1)  Microbial Metabolism                          56




            2)  Rate of Degradation                           59




        b.  Propylene Glycols                                 62




            1)  Microbial Metabolism                          62




            2)  Rate of Degradation                           64




        c.  Butylene Glycols                                  65




            1)  Microbial Metabolism                          65




            2)  Rate of Degradation                           66
                             iii

-------
                                                                   Page
         2.  Chemical Degradation
         3.  Transport Within and Between Media
         4.  Persistence and Bioaccumulation
     G.  Analytical Methods
III. BIOLOGICAL EFFECTS                                             71
     A.  Ethylene Glycol                                            71
         1    U                                                      71
         1.  Humans
             a.  Acute Toxicity
                                                                    73
             b.  Occupational Exposure
                                                                    73
                 1)  Aerosol Inhalation
                                                                    74
                 2)  Dermal Exposure
                                                                    74
             c.  Controlled Studies
         2.  Nonhuman Vertebrates
             a.  Absorption, Distribution, and Excretion
                 1).  Rat                                            75
                 2)  Rabbit                                         78
                 3)  Dog                                            78
                                                                    7 8
                 4)  Monkey
                                                                    81
             b.  Metabolism
                 1)  Rat                                            83
                 2)  Monkey                                          88
                                                                     89
             c.  Acute Toxicity
                                                                     89
                 1)  Oral  Administration
                                                                     89
                     a)  Lethal  Dose  Values
                                                                     91
                     b).  Signs
                                    iv

-------
B.
            c)   Tissue  and Organ Changes




            d)   Metabolic Acidosis




        2)   Dermal  and  Ocular Application




        3)   Parenteral  Administration




            a)   Lethal  Dose Values




            b)   Signs




            c)   Urine  Formation




            d)   Metabolic Acidosis




            e )   Hemat olo gy




    d.   Subacute Toxicity




        1)   Oral Administration




            a)   Renal  Changes




            b)   Calcium and Phosphorus Metabolism




            c)   Blood  Clotting




         2)   Inhalation Exposure




     e.   Chronic Toxicity




         1)   Oral Administration




         2)   Inhalation Exposure




         3)   Parenteral Exposure




     f.   Special Studies




         1)   Reproduction




         2)   Carcinogenicity




         3)  Mutagenicity




3.  Aquatic Organisms




4.  Plants




5 .  In Vitro Studies




Propylene Glycols




1.  1,2-Propanediol
   Page




   91




  92




  93




  93




  93




  95




  95



  96




  96




  97




  97




  97




  99




  99




 100




 103




 103




 104




 104




 105




 105




 106




 110




 110




 111




 111




113




113
                                  •v

-------
                                                    Page




a.  Humans                                          113




    1)  Acute Toxicity                              H3




        a)  Oral Ingestion                          H3




        b)  Dermal Exposure                         116




    2)  Controlled Studies                          H8




b.  Nonhuman Vertebrates                            H8




    1)  Absorption,  Distribution, and Excretion     118




    2)  Metabolism                                  122




    3)  Acute Toxicity                              I25




        a)  Oral Administration                     125




            ±.  Lethal Dose Values                  125




           ii.  Signs                               127




          iii.  Behavioral Effects                  128




        b).  Dermal, Ocular, and Ear Application     128




        c)  Inhalation Exposure                     129




        d)  Parenteral Administration               130




            i.  Lethal Dose Values                  130




           ii.  Signs                               130




          iii.  Hematology and Kemodynamics         134




    4)  Subacute Toxicity                           135




        a).  Oral Administration                     135




    5)  Chronic Toxicity                            136




        a)  Oral Administration                     136




            i.  Rats                                136




                i)  Weight Gain                     136




               ii).  Organ Effects                   138
                     VI

-------
                                                         Page




                  iii)   Hematology and Urinalyses        138



               ii.  Dogs                                 139




                    i)   Weight Gain                      139



                   ii)   Organ Effects                    14°



                  iii)   Hematology and Urinalyses        -*-^



            b)   Inhalation Exposure



                i.  Rats                                 142



               ii.  Monkeys



        6)  Special Studies                              l43



            a)   Reproduction                             -^3



                i.  Mice                                 143



               ii.  Rats                                 143 ,



              iii.  Hamsters                             143



               iv.  Rabbits                              144



                v.  Chickens                             144



            b)   Carcinogenicity                          14°



            c)   Mutagenicity                             149



                i.  Host Mediated Assay                  149



               ii.  Cytogenetic Assay



              iii.  Dominant Lethal Assay


                                                         IS?
    c.  Aquatic Organisms                                J~J^



    d.  Plants                                           152



    e.  In Vitro Studies                                 152



2.  1,3-Propanediol                                      154



    a.  Humans



    b.  Nonhuman Vertebrates
                        vii

-------
                                                            Page
            1)   Metabolism                                  154
            2)   Acute Toxicity
                a)   Oral Administration                     155
                    i.  Lethal Doses                        155
                   ii.  Glycogenic Action                   156
                b)   Parenteral Administration               156
            3)   Chronic Toxicity                            I57
            4)   Reproductive Studies                        157
C.  Butylene Glycols
    1.  1,2-Butanediol
        a.  Humans                                          161
        b .  Nonhuman Vertebrates                            161
            1)   Metabolism                                  161
            2).  Acute Toxicity                              162
                a)  Rats                                    162
                b).  Rabbits                                 163
                c)  Dogs                                    163
            3}  Subacute Toxicity                           163
        c.  Aquatic Organisms                               163
    2.  2,3-Butanediol                                      164
        a.  Humans                                          164
        b .  Nonhuman  Vertebrates                            164
            1)   Metabolism                                   164
            2)   Acute Toxicity                               164
            3)   Reproduction  Studies                         165
        c.  Aquatic Organisms                                165
    3.  1,3-Butanediol                                       165

                            viii

-------
                                                    Page




a.  Humans                                          165




    1)  Nutritional and Metabolic Studies           165




    2)  Dermal Toxicity                             169




b.  Nonhuman Vertebrates                            169




    1),  Metabolism                                  169




    2)  Acute Toxicity                              174




        a)  Lethal Dose Values                      174




        b).  Signs                                   174




        c)  Gluconeogenesis and Lipogenesis         177




        d)  Behavioral Effects                      178




    3)  Subacute Toxicity                           179




        a)  Rats                                    179




        b).  Cattle                                  181




        c).  Chickens                                181




    4)  Chronic Toxicity                            182




        a)  Rats                                    182




        b)  Dogs                                    182




    5)  Special Studies                             185




        a)  Reproduction                            185




            i.  Rats                                185




            ii.  Dogs                                190




          "iii.  Rabbits                             19°




            iv.  Chickens                            190




        b)  Mutagenicity                            192




            i.  Dominant  Lethal  Test                192




            ii.  Cytogenetic  Studies                 192

-------
                                                                 Page




                     c.  Aquatic Organisms                       192




                 4.  1,4-Butanediol                              193




                     a.  Humans                                  193




                     b.  Nonhuman Veterbrates                    193




                         1)  Metabolism                          193




                         2)  Acute Toxicity                      193




                             a)  Lethal Dose Values              193




                             b)  Signs                           193




                             c).  Dermal Effects                  197




                         3)  Subacute Toxicity                   197




                         4),  Reproduction Studies                198




                     c.  Aquatic Organisms                       198




IV.  REGULATIONS. STANDARDS. AND HANDLING                        199




     A.  Federal Regulations                                     199




         1.  Food and Drug Administration                        199




         2.  Environmental Protection Agency                     205




         3.  Occupational Safety and Health Administration       206




         4.  Department of Transportation                        206




     B.  State Regulations                                       206




         1.  Workplace                                           206




         2.  Food Contact                                        210




         3.  Water Quality                                       210




         4.  Air Emissions                                       210




     C.  Foreign Countries                                       210




         1.  United Kingdom                                      210




         2.  West Germany                                        211
                                  x

-------
                                                               Page




        3.  Japan                                               212




        4.  Canada                                              212




    D.  Other Standards                                          212




    E.  Handling and Storage Practices                           213




        1.  Handling, Storage,  and Transport                     213




        2.  Personnel Exposure                                   213




        3.  Accident Procedures                                 214




V.  EXPOSURE AND EFFECTS POTENTIAL                              215




TECHNICAL SUMMARY                                               218




BIBLIOGRAPHY                                                    229




CONCLUSIONS AND RECOMMENDATIONS                                 248




APPENDIX - Summary of Sources Employed                           249
                                 xi

-------
                                 PREFACE





     This report is a survey and summary of the literature available




through November 1978 on ethylene glycol, 1,2-propanediol, 1,3-pro-




panediol, 1,2-butanediol, 2,3-butanediol, 1,3-butanediol,  and 1,4-




butanediol.  Major aspects of properties, reactions,  production,




use, environmental exposure, biological effects and regulations are




reviewed.  A list of sources employed in the literature search appears




in the appendix.




     This document was prepared by the Franklin Research Center for




the Environmental Protection Agency under Contract 68-01-3893.
                                 XII

-------
                              LIST OF TABLES

Table                                                              Page

   1.  Nomenclature and Other Identifiers of Ethylene Glycol,         2
       Propylene Glycols, and Butylene Glycols

   2.  Physical Properties of Ethylene Glycol                        6

   3.  Physical Properties of Propylene Glycols                      7

   4.  Physical Properties of Butylene Glycols                       9

   5.  Specifications for Industrial Grade and Polyester Grade      11
       Ethylene Glycol

   6.  Specifications for Industrial Grade and U.S.P. 1,2-          12
       Propanediol

   7.  Specifications for Butylene Glycols                          13

   8.  U.S. Production and Sales of A) Ethylene Glycol and B).        22
       1,2-Propanediol

   9.  Ethylene Glycol Producers                                    24

  10.  1,2-Propanediol Producers                                    25

  11.  Butylene Glycol Producers                                    26

  12.  Imports of Ethylene Glycol into the United States            28

  13.  Imports of Propylene Glycol and Butylene Glycol into the     30
       United States

  14.  Market Price of the Glycols in the United States             32

  15.  Trade Names and Producers of Polyester Fiber                 34

  16.  Exports of U.S. Ethylene Glycol                              36

  17.  Exports of U.S. 1,2-Propanediol                              40

  18.  Composition of De-icers                                      46

  19.  Hazards of Ethylene Glycol Transportation                    49

  20.  Flow, Biological Oxygen Demand (.BOD) and Chemical Oxygen     51
       Demand (.COD) of Waste Streams Generated at Various Plants
       During Ethylene Glycol Manufacture

  21.  Biodegradation of Ethylene Glycol, Antifreeze and 1,2-Pro-    61
       panediol
                                 Xlll

-------
Table                                                            Page

 22.  Biodegradation of Ethylene Glycol and 1,2-Propanediol       63

 23.  Tissue Distribution and Excretion of ^C-Labeled Ethylene   76
      Glycol (.139 mg/kg, i.v.) in Rats

 24.  Urinary Excretion of 14C-Labeled Ethylene Glycol by Male    77
      Rats

 25.  Excretion and Tissue Distribution of lltC-Labeled Ethylene   80
      Glycol in Female Rhesus Monkeys

 26.  Urinary Excretion of Ethylene Glycol and lkC by Female      81
      Rhesus Monkeys

 27.  Effect of Pyrazole on Ethylene Glycol Toxicity and Meta-    85
      bolism in the Rat

 28.  Oral Lethal Dose Values for Ethylene Glycol                 90

 29.  Signs of Intoxication in Ten Dogs (Mixed Breed! Dying from  92
      Ingestion of Ethylene Glycol

 30.  Median Lethal Doses for Ethylene Glycol Administered Sub-   94
      cutaneously, Intravenously, or Intraperitoneally

 31.  Mortality in Animals Inhaling Ethylene Glycol Aerosol      101

 32.  Evaluation of Carcinogenicity in Texas-Yale Mice           107

 33.  Tumor Incidence in Fischer Rats Given Twice Weekly S.C. In-198
      jections of Ethylene Glycol for One Year

 34.  Level of 1,2-Propanediol Used in Food                      114

 35.  Amount of 1,2-Propanediol in the Blood, Saliva, and Urine  119
      o'f Three Subjects Following an Oral Dose of 1 ml/kg

 36.  Rate of Disappearance of 1,2-Propanediol in Dogs           121

 37.  Oral Lethal Dose Values for 1,2-Propanediol                126

 38.  Parenteral Lethal Dose Values for 1,2-Propanediol          131

 39.  Incidence of Neoplasms in Charles River Rats Fed 1,2-
      Propanediol in the Diet for Two Years

 40.  Number and Type of Tumors in Swiss Mice Treated with 1,2-  150
      Propanediol

 41.  Effect of 1,3-Propanediol on Mortality of White Leghorn    160
      Chicken Embryos
                                 xxv

-------
Table                                                            Page

 42.  Various Parameters Measured in the Blood of Volunteers     168
      Fed 1,3-Butanediol

 43.  Blood Levels of Acetoacetate, 3-Kydroxybutyrate, Pyruvate, 171
      Lactate, and Total Ketones in Rats Fed 1,3-Butanediol

 44.  Lethal Dose Values for 1,3-Butanediol                      175

 45 .  Summary of Reproduction and Lactation Responses in Re-     187
      productive Study in Rats:  Data on FQ Generation, Litterj
      Fed 1,3-Butanediol

 46.  Summary of Reproduction and Lactation Responses in Re-     189
      productive Study in Rats:  Data on FQ Generation, Litter2
      Fed 1,3-Butanediol

 47.  Lethal Dose Values for 1,4-Butanediol                      195

 48.  FDA Status of the Glycols Used as Food Additives           200

 49.  Regulations for Glycols Food Contact and Workplace         ..2.07
      Standards  in  Selected  States in Response to Queries

 50.  Water Standards for the Glycols in Selected States in      208
      Response to Queries

 51.  Air Standards for the  Glycols in Selected States in Re-    209
      sponse to Queries
                             LIST OF FIGURES

Figure                                                           Pa§,e

  1.  Production of Sthylene Oxide and Ethylene Glycol, Li-      15
      censed by Shell Development Co.

  2.  Manufacture of 1,2-Propanediol from Propylene Oxide        18

  3.  Pathway of Ethylene Glycol Metabolism in the Rat           82
                                 xv

-------
                            EXECUTIVE SUMMARY



     This report reviews aspects of production,  use,  environmental


exposure and biological effects of ethylene glycol,  two  isomers  of


propylene glycol (1,2- and 1,3-propanediol) and  four isomers  of  butylene


glycol (.1,3-, 1,4-, 2,3-, and 1,2-butanediol) .   About 3.7  billion


pounds of ethylene glycol were produced in 1977  in the U.S.  for  use


primarily in antifreeze and polyester fiber.  Domestic production of


1 j2-propanediol (.propylene glycol) is about 0.5  billion  pounds which


goes mainly into polyester resins, food, Pharmaceuticals,  and cello-


phane.  1,3-Propanediol is of only minor commercial importance.   Do-


mestic demand for 1,4-butanediol is 0.17 billion pounds  for  the  pro-


duction of tetrahydrofuran and acetylenic chemicals.   1,2- and 1,3-


butanediol are used to produce polymeric plasticizers while  the  2,3-


isomer is used as a solvent, humectant, and coupling agent;  production


figures are unavailable.


     Limited monitoring data make it difficult to adequately assess


environmental exposure to the glycols.  The major source of  contami-


nation by ethylene glycol and 1,2-propanediol is probably  from the


disposal of spent antifreeze and de-icing fluids.  The glycols are


capable of being degraded by a variety of acclimated and unacclimated


soil, water, and sewage microorganisms.  Under different testing


conditions, complete degradation usually occurred within 3-20 days.
                     a

     In humans, ethylene glycol intoxication, usually as a result of


accidental ingestion of antifreeze, may result in nausea,  hypertension,


tachycardia, cardiopulmonary failure, renal impairment,  and coma.


The oral lethal dose in humans is about 1.4-1.6  g/kg and in laboratory
                                 xvi

-------
animals, 6-15 g/kg.  This toxicity is due to a metabolite of ethylene




glycol, glycolate.  Microscopic findings in humans and laboratory




animals often include deposition of calcium oxalate crystals in the




kidney.  Significant inhalation exposure is likely only when the gly-




col is at elevated temperatures.  Only one occupationally-related




case report is available in the literature involving ethylene glycol




inhalation.  Intermittent exposure to 10 or 57 mg/m3 for 90 days




was without effect to several laboratory animals while continuous




exposure to 12 mg/m3 caused eye irritation.  Continuous exposure to




256 mg/m3 of monkeys resulted in some impairment of sensory discrimi-




nation.   No carcinogenic potential has been noted in mice given a




single s.c. injection, in mice by lifetime skin painting, or in rats




given  twice weekly s.c. injections for one year.




     1,2-Propanediol is a generally-recognized-as-safe CGRAS) food




additive.  In laboratory animals, the oral LDsQ ranges between about




14-30  g/kg.  Large doses result in a loss of equilibrium, central




nervous system depression and respiratory failure.  No consistent




chronic effects were noted in rats fed up to 10% in the diet or dogs




fed up to 20%.  No teratogenic effects were noted in mice, rats,




hamsters, or rabbits given 1,2-propanediol during gestation.  1,2-




Propanediol was not carcinogenic when administered by single or mul-




tiple  injection, by chronic dietary addition or by lifetime skin




painting.  No mutagenic effects were detected in a host-mediated




assay, cytogenetic assay, or dominant lethal gene test.  •




     In one study, 1,3-propanediol was twice as toxic as 1,2-propane-




diol by subacute dietary feeding.
                               xvii

-------
     1,3-Butanediol has been extensively studied as a synthetic




metabolizable source of dietary energy.  The oral LD__ in mice and




rats ranges from 23 to 30 g/kg.  At large doses, narcosis results from




muscle relaxation.  Some investigators reported a decrease in body weight




gain at high levels of 1,3-butanediol in the diet  (often 20% or more)




but no histological changes have been attributed to this diol.  A




gradual decrease in reproductive rate was noted in a multigeneration




study in  rats fed up to 24% in the diet.  No teratogenic effects were




observed  in litters from rats, dogs, or rabbits given butanediol prior




to and/or during pregnancy.  Furthermore, no adverse effects were noted in




a dominant lethal test and in a cytogenetic test in rats.




     Few  studies are available on 1,2- , 2,3-  , and 1,4-butanediol.  In




laboratory animals, the oral LD,... averages 16  g/kg, 8,9 g/kg, and 1.2-2.5




g/kg, respectively, for these isomers; the 1,4- isomer is the more acutely




toxic isomer, acting as a central nervous system depressant.
                                xvin

-------
                   I.  PHYSICAL AND CHEMICAL DATA
A.  Chemical Structure


     Glycols are compounds containing two hydroxyl groups (—OH) at-

tached to carbon atoms in an aliphatic carbon chain.  The glycols

discussed in this report have the following structures:
     1,2-Ethanediol
     (.ethylene glycol)
     1,2-Propanediol
     (propylene glycol)
     1,3-Propanediol
     1,3-Butanediol
     1,4-Butanediol
     2,3-Butanediol
     1,2-Butanediol
CH2CH2

OH OH
   OH OH

CH2CH2CH2

OH    OH


CH3CHCH2CH2

   OH   OH


CH2CH2CH2CH2

OH       OH


CH3CH-CHCH3

   OH OH


CH2CHCH2CH3

OH OH
Synonyms and other identifiers of these  glycols  appear  in  Table  1.

The  term propylene glycol is often  used  in  the literature  to  refer  to

1,2-propanediol; the latter term is used throughout  this report.

-------
                                                        Table 1
                            Nomenclature and Other Identifiers of Ethylene  GlycoL, Propylene Glycols,
                                                 and Butylene Glycols
Chemical Abstracts
Service (CAS)
10th Collective
Index Name
CAS
Registry
No.
EPA Toxic
Substances
„ . , . . Synonyms
Control Act J }
List No .
Molecular
Formula
Wiswesser
Line
Notation
          ethylene  glycol
ro
           1,2-propanediol
107-21-1     A806-1653     1,2-dihydroxyethane     C2H602
                           1,2-ethanediol
                           ethane-1,2-diol
                           ethylene alcohol
                           ethylene dihydrate
                           glycol
                           glycol alcohol
                           monoethylene glycol
57-55-6      B450-5740     propylene glycol        C3H802
                           methylethylene glycol
                           methyl glycol
                           monopropylene glycol
                           propane-l,2-diol
                           alpha-propylene glycol
                           l,2<-propylene glycol
                           trimethyl glycol
                           1,2-d ihydroxyp rop ane
                           sirlene
Q2Q
QY1Q

-------
                                        Table  I  (Continued)
Chemical Abstracts
Service (CAS)
10th Collective
Index Name
CAS
Registry
No.
EPA Toxic
Substances
„ . , . Synonyms
Control Act
List No .
Molecular
Formula
Wiswesser
Line
Notation
1,3-p ropanediol
1,3-butanediol
1,4-butanediol
                          2,3-propanediol
                          2-hydroxypropanol
504-63-2    B451-0531     1,3-dihydroxypropane    C3He02
                          trimethylene glycol
                          1,3 propylene glycol
                          Beta-propylene glycol
                          2-deoxyglycerol
107-88-0    A322-3373     Beta-butylene glycol    C^E-iQ02
                          1,3-butylene glycol
                          1,3-dihydroxybutane
                          methyl trimethylene glycol
                          butane-1,3-diol
110-63-4    A322-3510     1,4-butylene glycol     CttH1002
                          1,4-dihydroxybutane
                          1,4-tetramethylene glycol
                          tetramethylene glycol
                          butane^l,4-diol
Q3Q
QY2Q
Q4Q

-------
                                        Table 1 (Continued)
Chemical Abstracts
Service (CAS)
10th Collective

Index Name
CAS
Registry
No

EPA Toxic
Substances _
^ t i A *. Synonyms
Control Act J

List No.
Molecular
Formula


Wiswesser
Line
Notation

2,3-butanediol
1,2-butanediol
513-85-9,   A322-3843
35007-63-7,
6982-25-8,
24347-53-8,
5341-95-7,
19132-06-0
584-03-2
A322-3155
2,3-dihydroxybutane
dimethylene glycol
2,3-butylene glycol
pseudobutylene glycol
sym-dimethylene glycol
1,2-butylene glycol
1,2-dihydroxybutane
ethyl ethylene glycol
                                                    QY,YQ

-------
B.  Properties of the Pure Material





     Ethylene glycol is a colorless, practically odorless,  relatively




involatile, hygroscopic liquid.  Some of its important physical pro-




perties are listed in Table 2.  Ethylene glycol is miscible in all




proportions with water and the lower aliphatic alcohols,  aldehydes,




and ketones, but is almost insoluble in hydrocarbons (Curme and




Johnston, 1952) .  It is an excellent permanent antifreeze because  it




is miscible with water and has such a low freezing point.  Ethylene




glycol is considered a slight fire hazard when exposed to heat or




flame and a slight explosion hazard when exposed to flame CSax, 1975} .




The lower flammable limit is 3.2% (Table 21.




     1,2-Propanediol is a colorless, odorless, slightly sweet tasting




liquid.  As it  contains an asymmetric carbon atom, it occurs in two




optically active forms (levorotatory and dextrorotatory isomers).




Data on physical properties, which appear in Table 3, were determined




on the racemic mixture.  1,2-Propanediol is completely miscible with




water and most lower molecular weight aliphatic compounds containing




oxygen or nitrogen; most aromatic and halogenated compounds are only




partly miscible in 1,2-propanediol (.Curme and Johnston, 1952).  It is




considered a moderate fire hazard when exposed to heat or flame and




a moderate fire hazard when exposed to flame  (Sax, 1975) .




     1,3-Propanediol is a viscous, colorless, hydroscopic, odorless




liquid possessing a brackish irritating taste.  It is soluble  in




water, alcohol, and ether (.Curme and Johnston, 1952).  Physical pro-




perties of 1,3-propanediol are listed in Table 3.

-------
                            Table 2
Physical Properties of Ethylene Glycol (Dow, 1978a, Union Carbide,
    1978 and 1976; Perl et al., 1976; Curme and Johnston, 1952)
        Property
Data
        appearance


        boiling point, 760 mm Hg

        density, 20°C

        flash point

        flammable, limits

        freezing point, 760 mm Eg

        heat of combustion, 20°C

        molecular weight

        saturated vapor at 20°C

        solubility in water at 20°C

        vapor  density (air =1)

        vapor  pressure at 20°C

        refractive index, n^

        %  volatiles  by volume

        weight/U.S.  gallon, 20°C

        conversion factors
         (25° C; 760  mm Hg)

        olive  oil/water partition
         coefficient, 37°C
colorless liquid
mild odor
197.6°C

1.11336 g/ml

241°F

lower 3.2%

-12.7°C

-283.3 kcal/mol

62.07

79 ppm (204 mg/m3)

complete

2.14

0.06 mm Hg

1.4318

Nil

9.28 Ibs

1 ppm = 2.74 mg/m3
1 mg/fc = 394 ppm

34 x 10-1*

-------
                            Table 3
Physical Properties of Propylene Glycols (Dow,  1978b;  Union Car-
        bide, 1978; Dow, 1966; Perl et al., 1976; Curme
                      and Johnston, 1952}
Property
                           1,2-Propanediol
                                                     1,3-Propanediol
appearance


boiling point, 760 mm Eg

density (20°C), g/ml

flash point

flammable limits


freezing point, 760 mm Hg

heat of combusion, 20°C

molecular weight

refractive index n^°

solubility in water at 20°C

vapor density  (air =1)

vapor pressure at 25°C

saturated vapor at 20°C


weight/U.S. gallon, 25°C

conversion factors
(25° C, 760 mm Hg)

olive oil/water Partition
coefficient, 37°C
                           colorless liquid     colorless  liquid
                           187.3°C

                           1.0363

                           214 °F

                           lower:  2.6%;
                           upper:  12.5%

                           -60°C

                           435.9 kcal/mol

                           76.10

                           1.4326

                           complete

                           2.62

                           0.22 mm Eg

                           299 ppm
                           C917 mg/m3)

                           8.62

                           1 ppm = 3.11 mg/m
                           1 mg/H = 322 ppm

                           rt o   1 r*—4
                           23 x 10
                                                     210-211°C

                                                     1.0597
                                                     76.10



                                                     complete

                                                     2.6
                                                     1 ppm = 3.11 mg/nr
                                                     1 mg/£ = 322 ppm

                                                     24 x 10"4

-------
     There are four butanediols:  1,3-, 1,4-, 2,3-, and 1,2-butanediol.




Physical properties of these isomers are listed in Table 4.  Butane-




diols are usually viscous, colorless, odorless liquids which are




soluble in water.  1,3-Butanediol is also soluble in dibutylphthalate,




methyl ethyl ketone and other solvents, but insoluble in straight




chain hydrocarbons (Wagner, 1966).  1,4-Butanediol is completely




soluble in water, methanol, ethanol, and acetone and moderately solu-




ble in acetate and only slightly soluble in hydrocarbons (GAF, n.d.).




The meso form of 2,3-butanediol is, moderately soluble in diisopropyl




ether, while the d,l form is very soluble in this ether (Windholz, 1976)




1,3-Butanediol has been classed as a slight fire hazard, while the




other butanediols present moderate fire hazards when exposed to heat




or flame (Sax, 1975) .









C.  Properties of the  Commercial Material





     Specifications for industrial grade (99.0% pure) and polyester




grade ethylene glycol  are listed in Table 5.  Impurities in both




grades include acetic  acid, ash, and diethylene glycol.




     Specifications for U.S.?  (99.5% pure) and industrial grade 1,2-




propanediol appear in  Table 6.  Both grades contain acetic acid.




     1,4-Butanediol is available in a technical (97% pure) or an




anhydrous  grade  (98.5% pure); the technical grade  contains a maximum




of 0.4% water while the anhydrous grade contains 0.04% water (Freifeld




and Hort,  1966).  Specifications are shown in Table 7.  Impurities




include butenediol and butyrolactone .

-------
                                Table 4
Physical
Sax,

appearance
boiling point
CD
Cd)
Cm)
Cd,l)
density, 20 °C
CD
Cd)
(m)
Cd,l)
Properties of Butylene Glycols CPerl et al . , 1976;
1975; Weast, 1974; Budke and Banerjee, 1971;
Wagner, 1966; Freifeld and Hort, 1966)
1,3-Bu-
tanediol
colorless
liquid
, °C
107-110
204
—
207.5

1.005
1.0053
—
1.0053
molecular weight 90.12
flash point
250°F
refractive index
CD
Cd)
(m)
Cd,l)
solubility in
'l.4418
—
1.4410
complete
1,4-Bu- 2,3-Bu-
tanediol tanediol
colorless yisco"s
, . . , liquid
liquid ^ .
or solid
221-231
178
180-182
182
177
1.0171
0.9869
0.9872
1.0003
1.0003
90.12 90.12
>250°F 185°C
Copen cup) (open cup)
1.4446
1.4318
1.4306
1.4325
—
complete complete
1,2-Bu-
tanediol
colorless
liquid
__
94-96
192-194
—
190.5

—
1.0059
—
1.0024
90.12
104 °F
—
1.4375
—
1.4378
complete
water

-------
                           Table 4 (Continued)
conversion factors
(25° C, 760 mm Hg)
1,3-Bu-
tanediol
1 ppm =
3.68 mg/m3
272 ppm
1,4-Bu-
tanediol
1 ppm =
3.68 mg/m3
1 mg/£ =
272 ppm
2,3-Bu-
tanediol
1 ppm =
3.68 mg/m3
1 ing/?. =
272 ppm
1,2-Bu-
tanediol
1 ppm =
3.68 mg/nr
1 mg/£ =
272 ppm
olive oil/water
Partition coef-
ficient, 37°C

vapor pressure
saturated vapor
at 20°C
              84 x 10"
0.06 mm Hg
at 20°C

79 ppm
C296 mg/m3).
     The specifications for commercially available 1,3- and 2,3-

butanediols are listed in Table 7.



D.   Chemical Reactions Involved in Use


     The most important commercial reaction of the glycols is esteri-

fication (Miller, 1966).  Esterification is carried out with organic

and  inorganic acids, acid halides, or acid anhydrides.  The reaction

of a glycol with monocarboxylic acids, such as acetic acid, produces
                                    10

-------
                                 Table 5
           Specifications for Industrial Grade and Polyester
               Grade Ethylene Glycol (Union Carbide, 1978)
                                  Industrial Grade
                     Polyester Grade
purity, % by weight, minimum

acidity, % by weight,
maximum as acetic acid

water, % by weight, maxi-
mum

ash, % by weight, maximum

odor
specific gravity at 20/20°C

suspended matter


water solubility
diethylene glycol, % by
weight, maximum

iron, ppm maximum
99.0

0.005


0.2


0.005

mild


1.1151-1.1156

substantially
free

miscible in all
proportions at
25°C

0.5
0.005
0.08
0.005

mild, practi-
cally none

1.1151-1.1156

substantially
free

miscible in all
proportions at
25°C

0.08
                     0.07
primarily mono- or diesters depending on the molar ratios of the re-

actants.  An example of this is the reaction of ethylene glycol with

acetic acid:
                                    11

-------
                                 Table 6
             Specifications for Industrial Grade and U.S.P.
                  1,2-Propanediol (Dow,1975, 1978b)
                                  Industrial Grade
                     U.S.P.
purity ,  % by weight ,  minimum

acidity, ppm maximum as
  acetic acid

water, % by weight, maximum

ash, % maximum, sulfated

odor


taste


specific gravity 20/20°C

suspended matter


water solubility

chlorides, ppm maximum

heavy metals, ppm maximum
  as lead

arsenic, ppm maximum
  at
20
0.2
characteristic
odor
1.0377-1.0390
complete

1
99.5

20


0.2

.005

practically
none

slight
characteristic

1.0376-1.0389

substantially
free

complete

1

5
 iron, ppm maximum
0.5
0.3
                                    12

-------
                                 Table 7
                   Specifications for Butylene Glycols
                   l,4-Butanediole
                   CGAF, n.d.;
                   Freifeld and
                   Eort, 1966)
                   1,3-Butanediol
                   CBudke  and
                   Banerjee,  1971;
                   Celanese,  1977)
                2,3-Butanediol
               (Budke and Baner-
                jee,  1971;  Curme
                and Johnston,  1952)
purity, %, mini-
  mum

color, APHA
  maximum

appearance at
  25°C

boiling point

solidification
  point, °C,
  minimum

water content,
  % maximum

solidification
point, °C,
  minimum
   Cb)
   20
clear liquid
99.0
   19.0
   (a)
   19
carbonyl number,      1.5
  mg KOH/g, maximum

distillation range,   —
  760 torr

acidity, as ace-
  tic acid, %
  maximum

specific gravity
  20/20°C

butenediol (.%)        <0.05

butyrolactone C%1     0.1
0.50
                   200-215°C
                      0.005
                   1.004-1.006
                                        182°C
                                              Cc)
                   174-186°C
                   0.01
                   1.006-1.010
     Two grades are available, regular (0.4% water, maximum) and anhy-
droug (contains less than 0.04% water).

     Technical grade:  97% pure anhydrous grade:  98.5% pure.
    °Mixture of 85% meso and 15% dextro-levo isomers.
                                    13

-------
     HOCH2CH2OH + CH3COOH -*• CH3COOCH2CH2OH + H20




                            ethylene glycol monoacetate




     HOCH2CH2OH +  2CH3COOH -»• CH3COOCH2CH2OOCCH3 + 2H20




                            ethylene glycol diacetate




Cyclic or linear polyesters are formed by the reaction with dicarboxylic




acids, an example of which is given below:




     HOGH2CH2OH + HOOCRCOOH -»• (OCH2CH2OOCRCO).  + 2H20





Commercial applications of low-molecular weight organic-acid esters




of ethylene glycol include use in solvents.  Glycol esters of acrylic




and methacrylic acids are used to prepare polyacrylate resins.  The




terephthalate ester of ethylene glycol is used to make polyester fiber.




Fatty acid esters of propylene glycol are used in cosmetics and pharma-




ceutical preparations.  Polyesters of 1,3-butanediol are made by re-




action with oxalic, succinic, glutaric, or adipic acids or with maleic




or phthalic anhydrides.  Esters of 1,4-butanediol are used in thermo-




plastic polymers; polyesters are used as plasticizers (Miller, 1966;




Wagner, 1966; Freifeld and Hort, 1966).




     Another commercially important reaction of the glycols is etheri-




fication.  An example is heating ethylene glycol with a dehydration




catalyst (H2SOit) to form dioxane, a cyclic diether.




     Acetals and ketals of glycols are formed by condensation reac-




tions with appropriate aldehydes and ketones.  These have applications




as solvents.
                                    14

-------
                II.  ENVIRONMENTAL EXPOSURE FACTORS


     Aspects of manufacture, use, waste, handling, environmental con-

tamination, degradation, and analytical detection methods are con-

sidered in the following sections.



A.  Production

     1.  Production Processes

          a.  Ethylene Glycol


     Two methods are currently used in the U.S. to manufacture ethylene

glycol.  The more widely used process involves the hydration of ethylene

oxide.  A method for the direct production of ethylene glycol from

ethylene was recently put into commercial operation by Oxirane Cor-

poration .

     Figure 1 shows a flow chart for the production of ethylene glycol

from ethyletie oxide.  In a widely used method of producing ethylene

oxide, ethylene and oxygen are fed into an isothermal catalytic
        Figure 1.  Production of ethylene oxide and ethylene
                   glycol, licensed by Shell Development Co.
                   (Anon,, 1977a).   Reprinted by permission
                   from Gulf Publishing Co., Houston,  Texas

                                    15

-------
reactor; the selective silver catalyst highly favors ethylene oxide




formation.  About 60-70% of the ethylene is converted to ethylene




oxide at reaction temperatures of 270-290°C.  The reactor effluent




gas containing the ethylene oxide product is cooled, then the ethylene




oxide is recovered by absorption in water; unabsorbed gases are re-




cycled.  Recycle gas is scrubbed to remove excess C02.  The ethylene




oxide is stripped from the water, then distilled to remove light




ends.  To produce ethylene glycol, ethylene oxide is then hydrated




in acid or by pressure in a tower reactor.  Acid hydration requires a




30 minute contact at 50°-70°C In a 0.5-1.0% sulfuric acid solution.




In pressure hydration, 185 psi and a contact time of one hour at




195°C are required.  Using either hydration process, diethylene glycol




and triethylene glycol are formed as by-products, which can be se-




parated from ethylene glycol by vacuum distillation ('Anon, 1977a  & b;




Lowenheim and Moran, 1975) .




     A new process of manufacturing ethylene glycol directly from




ethylene by acetoxylation was developed by Halcon International, Inc.




and put into commercial use at the Oxirane plant in Channelview,




Texas,  which came on stream in early 1978  (Anon., 1977c)-   The




first step in the Halcon process is the catalyzed liquid-phase oxida-




tion of ethylene in acetic acid to a mixture of mono- and diacetates




of ethylene glycol.  The acetates are then hydrolyzed to ethylene




glycol at 1Q7-130°C and 1.17 atm pressure; acetic acid is recycled




(Hatch and Matar, 1978a).  The overall ethylene glycol yield by this




process is 95%, compared to 67% via ethylene oxide hydration (Brown-




stein, 1974) .
                                    16

-------
     Union Carbide has obtained a patent on the production of ethylene




glycol from synthesis gas.  If this process were commercialized, ethylene




glycol could be manufactured from a variety of raw materials; produc-




tion would then be independent of natural gas and petroleum supply




(Hatch and Matar, 1977).  The reaction is carried out at 200°C and




8,000 psi and is as follows:




                     3CO + 5H2 -*• HOCH2-CR2OH + CH3OH.




Methanol is a by-product, the amount depending on the reaction condi-




tions.  This process is predicted to be of major importance in the mid-




1980's (Hatch and Matar, 1978a).




     Three other processes to produce ethylene glycol have been em-




ployed commercially in  the past:  a) the reaction of carbon monoxide




and formaldehyde; this method was discontinued in 1968 by duPont;




b) the fermentation of molasses was a not widely used method which




was discontinued in the early 1970's; c) vapor-phase oxidation of




propane; this was discontinued by Calanese in 1975 (Lowenheim and




Moran, 1975) .








          b.  Propylene Glycols





     1,2-Propanediol is manufactured commercially by the hydrolysis




of propylene oxide.  This process, shown in Figure 2, is analogous to




that used to produce ethylene glycol from ethylene oxide (Lowenheim




and Moran, 1975).  Propylene oxide yields 1,2-propanediol by acid




or pressure hydration in a tower reactor.  Hydration occurs at ele-




vated temperatures (EPA, 1975) .  Reaction products are then passed




from the reactor to evaporation and drying towers where excess water
                                    17

-------
CATALYST-

WATER —
 PROPYLENE
 6LYCOL «—
PROPYLENE
OXIDE          >
REACTOR
              a. e9 u- i—
                            STEAM JETS
                               AND
                            BAROMETRIC
                            CONDENSERS
                            WASTEWATER
                                  CRUDE
                                  DI-PROPYLENE
                                  GLYCOL
                                  STORAGE
                                                                        STEAM
                                                    TO ATMOSPHERE
                                                  LIGHT  ENDS
                                                  TO  INCINERATION
                                                                *FOOO GRADES
                                                                  DI-PROPYLENE GLYCOL
                                                   HEAVY ENDS
                                                   TO INCINERATION
    Figure 2.  Manufacture of 1,2-Propanediol from propylene oxide  (EPA,  1975),




    is removed.  Crude 1,2-propanediol is then passed to  a  series of  frac-

    tionators where water and light ends are removed; then  industrial grade

    1,2-propanediol is separated and condensed.  Dipropylene glycol is ob-

    tained as a by-product.

         1,3-Propanediol is only of minor commercial  importance.  It  is
                                        18

-------
primarily recovered in commercial glycerol plants only when there is




enough demand.  1,3-Propanediol has also been prepared by heating




Y .Y-dihydroxydipropyl ether with hydrobromic acid then hydrolyzing




the reaction product with sodium hydroxide (Budke and Banerjee,  1971) .








          c.  Butylene Glycols





     1,4-Butanediol is manufactured at GAF and BASF by the condensa-




tion of acetylene with formaldehyde via cupric acetylide catalysts




(Brownstein and List, 1977).  The copper acetylide is formed by  the




reaction of acetylene with copper salts such as ammonical cuprous




hydroxide.  Reaction of formaldehyde with copper acetylide under 5-15




atm. pressure at 90-100°C gives the butynediol in 95% yield.  The




butynediol  is hydrogenated over Raney nickel catalysts at 40-120°C




and 75-300 psig to give, a 85-90% yield of 1,4-butanediol.  Alternate




feedstocks  for 1,4-butanediol, including maleic anhydride, propylene,




and .butadiene have been proposed but have not been commercialized in




the U.S.




     The major commercial method for manufacturing 1,3-butanediol is




the catalytic hydrogenation of acetaldol (Wagner, 1966).  Catalysts




used include Raney nickel, copper, and platinum oxide.  Temperatures of




65-12Q°C and a pressure of 700 psi are employed.  From the hydrogena-




tion product, low boiling materials are removed; these include ethyl




alcohol, water, and n-butanol.  The reaction product is then filtered




to remove precipitated salts and catalyst and is redistilled to purify




the 1,3-butanediol.




     Various bacteria produce 2,3-butanediol through fermentation and




have been used in the commercial preparation of this glycol.  Vergnaud
                                    19

-------
(1950) described in a patent the process of making 2,3-butanediol by


fermentation of sugar or molasses using bacteria of the genus Mesen-


tericus.  2,3-Butanediol can also be produced by fermentation of


potatoes, wheat mash, or enzyme mash (Curme and Johnston,  1952).


The proportion of optical isomers produced is a function of the bacteria


used.  Fermentation products depend on the strain of bacteria employed;


these products may include (besides 2,3-butanediol). lactic acid, ethanol,


glycerol, formic acid, hydrogen, and carbon dioxide.  2,3-Butanediol


is recovered by distillation, solvent extraction, dialysis or steam


stripping.  2,3-Butanediol can also be made from gaseous mixtures


containing isobutylene and normal butenes; this mixture is combined


with hydrogen peroxide and formic acid; then, the reaction product


is distilled to remove fractions containing 2,3- and 1,2-butanediols


(Cosby, 1957).  Keith et al. (1961) described the preparation of


2,3-butanediol in 55% yield by oxidation of 2-butene.


     1,2-Butanediol is produced from butylene oxide (Hatch


and Matar, 1978b) .  The oxide is produced from 1-butene by chloro-


hydrination with hydrochlorous acid, followed by epoxidation.  Buty-


lene oxide is then hydrolyzed to 1,2-butanediol, as shown below:


                    0

                   /\        H+
          CH3-CH2-CH-CE2 + H20 -»• CH3-CH.2-CHOH-Ctt2OH.






     2.  Quantity Produced



     Production of ethylene glycol is the U.S. during 1977 was 3.7


billion pounds.  This represents an 80% increase in production in


the last decade, during which time production increased from 2.0
                                    20

-------
billion pounds in 1968; production peaked at 3.8 billion pounds during




1972 and 1975 (Table 8).  Ethylene glycol production during January-




October 1978 was 3.25 billion pounds (U.S. Int. Trade Commission, 1978)




and year end totals are expected to exceed 3.7 billion pounds (Anon,




1977d).  Production is currently less than 60% of the in-place design




capacity of manufacturing units (Anon, 1977d)..




     Total domestic production of 1,2-propanediol was about Q.5 billion




pounds during 1977, an increase of .40% during the last decade; during




this time, maximum production occurred in 1972 and 1976 (Table 8).




     The U.S. International Trade Commission does not list production




figures for 1,3-propanediol or for the butanediols.   Estimates of plant




capacity are discussed in the next section.








     3.  Domestic Producers and Production Sites





     There are 12 producers of ethylene glycol in the United States.




Based on announced capacities and trade estimates, the total capacity




for 1978 is 6.7 billion pounds (Table 9).  More than one billion pounds




of capacity were added during 1977 and a 0.8 billion pound capacity




plant  (Oxirane) was added during 1978 (Anon, 1977d) .  The largest




producer of ethylene glycol is Union Carbide, with a total capacity




of about 2.3 billion pounds.  Of the 16 sites where ethylene glycol




is produced, 7 are in Texas, 5 in Louisiana, 2 in Puerto Rico, 1 in




Kentucky, and 1 in Illinois.  ICI Americas has announced construction




of an ethylene glycol/ethylene oxide plant "at a Texas site" which




will be completed in 1981 (Anon, 1978a) .  Another ethylene glycol/




ethylene oxide plant will be built in Texas City, Texas; Union Carbide
                                    21

-------
Table 8
U.S. Production and Sales of A) Ethylene Glycol and B) 1,2-Pro-
panediol (U.S. International Trade Commission 1973-
1978; U.S. Tariff Commission 1968-1972).

A. Ethylene Glycol
1978
1977
1976
1975
1974
1973
1972
1971
1970
1969
1968
B. 1,2-Propanediol
1978
1977
1976
1975
1974
1973
Production
(.thousand
pounds)

(3,252,824)a
3,675,461
3,334,587
3,809,003
3,340,695
3,277,639
3-.761.W3
3,070,007
3,037,501
2,570,947
2,042,846

(345,760)_a
489,064
516,932
390,836
510,206
501,808
Sales
(thousand
pounds)

—
2,958,366
2,525,135
2,847,976
2,699,075
2,828,598
3,113,931
2,630,826
2,209,675
1,936,013
1,627,336

—
487,152
469,850
365,513
476,450
520,298
Unit Value of Sales,
cents per
pound

—
0.19
0.2Q
0.21
0.14
0.07
0.06
0.06
0.07
0.06
0.07

—
0.25
0.26
0.27
0.21
0.09
   22

-------
                           Table 8 (Continued)
Production Sales Unit Value of Sales,
Cthousand (.thousand cents per
pounds) pounds). pound
1972
1971
1970
1969
1968
562,583 538,037 0.08
421,446 427,311 Q.09
428,181 395,459 0.09
460,565 411,173 0.09
352,876 333,055 0.09
     Preliminary; production from January 1-October, 1978.
has announced construction of a 700 million pound/year ethylene glycol




facility there by 1981 (Anon, 1977e)..




     There are five domestic producers of 1,2-propanediol with a total




capacity of 0.8 billion pounds (Table 10).  The largest producers are




Dow Chemical (0.40 billion pounds) and Oxirane Chemical Co. (0.25




billion pounds).  Production is at six sites:  three in Texas, and




one each in Louisiana, Kentucky, and West Virginia.  The limiting




factor for production of 1,2-propanediol is the availability of pro-




pylene oxide; expansions of propylene oxide capacity have been an-




nounced by Dow and Oxirane (Anon, 1977f) .




     1,3-Propanediol is manufactured by the Guardian Chemical Cor-




poration (Eastern Chemical Division), in Hauppauge, N. Y. (SRI, 1975);




the capacity of this plant is not available.
                                    23

-------
                      Table 9
Ethylene Glycol Producers (MCP, 1977; Anon, 1977e)
                            Capacity
       Producer             (millions of
                            pounds per year)
   BASF Wyandotte Corp.
     Geismar, Louisiana         250

   Calcasieu Chemical Corp.
     Lake Charles, Louisiana    230

   Celanese Corp.
     Clear Lake, Texas          500

   Dow Chemical USA
     Freeport, Texas            330
     Plaquemine, Louisiana      350
                     «a
   Jefferson Chemical
     Port Neches, Texas         360

   Northern Petrochemical
     Joliet, Illinois           350

   Olin Corp .
     Brandenburg, Kentucky       50

   Oxirane
     Channelview, Texas         800

   PPG Indust., Inc.
     Beaumont, Texas            200
     Guaynilla, Puerto Rico     400

   Shell Chemical Co.
     Geismar, Louisiana         200

   Texas Eastman
     Longview, Texas            180

   Union Carbide Corp.
     Ponce, Puerto Rico         730
     Seadrift, Texas            870
     Taft, Louisiana            700

            Total             6,700
        Capacities are probably overstated
   and include total glycols, for example,
   triethylene glycol.
                          24

-------
                                Table 10
                 1,2-Propanediol Producers (Anon, 1977f)
                       Producer
Capacity
(millions of
pounds per year)
                Dow Chemical
                  Freeport, Texas             250
                  Plaquemine, Louisiana       150

                Jefferson Chemical Co.
                  Port Neches, Texas           40

                Olin Corp.
                  Brandenburg, Kentucky        45

                Oxirane Chemical Co.
                  Bayport, Texas              250

                Union Carbide Corp .
                  S. Charleston, West         100
                  Virginia                   	
                             Total            835
     Production data for butylene glycols are listed in Table 11.

Producers of 1,4-butanediol are duPont, GAF Corp. and BASF Wyandotte

Corp.  Total capacity is in excess of 0.2 billion pounds (.figures

for GAF are unavailable) at two sites in Texas, one in New Jersey

and one in Louisiana.  The U.S. International Trade Commission (1977)

lists Celanese Corp. and duPont as producers of 1,2- and 1,3-butanediols;

plant capacities have not been disclosed.  Optically active  (D)(-)
                                    25

-------
                             Table  11
   Butylene Glycol  Producers  (SRI, 1975; Anon,  1976a;  1977g; U.S,
                       Int. Trade  Comm., 1977).
            Producer
Capacity
(millions of
pounds per year
  I.  1,4-Butanediol

        E.  I. duPont de  Nemours  and  Co.,  Inc.
         LaPorte,  Texas

        GAF Corp.
          Calvert  City, Kentucky
         Linden,  New Jersey
         Texas  City, Texas

        BASF Wyandotte  Corp.
         Geismar,  La.

 II.  1,2-  and 1,3-Butanediols

        Celanese  Corp.
         Bishop Texas

        E.  I. duPont  de Nemours and Co.

III.  2,3-Butanediol

        Burdick  and Jackson Labs.
         Subsidiary  of Hoffman
         LaRoche,  Inc.
         Muskegon, Mich.
162, estimated

capacity not dis-
closed
55
capacity not dis-
closed
capacity not dis-
closed
      Q
       Capacity figures  are  not  available  from duPont;  this  esti-
  mate of 162  million  pounds is  based  on an  estimate  of 90 million
  pound capacity in 1976 plus 80%  of 90 million pounds;  duPont
  announced in 1976* an 80% increase  in capacity by 1978 (Anon,
  1976a).
                                  26

-------
2,3-butanediol is manufactured by Hoffmann-LaRoche; total capacity has




not been disclosed.







     4.  Imports and Foreign Producers





     Imports of ethylene glycol into the U.S. during 1976 totalled about




45 million pounds and came primarily from Japan (36.2%)  and the Nether-




lands (.34.7%); other sources included Belgium, Canada, the United King-




dom, and West Germany (Table 12).  As shown in Table 12, total imports




since 1972 have been quite variable.  A peak level was reached during




1975 when imports exceeded 371 million pounds, coming  from Belgium




(.32.5%), Japan (30.2%), and the Netherlands (23.1) among other countries.




In 1973, only 23 billion pounds were imported.




     According to a study by Monkman-Rumsey on worldwide ethylene oxide




supply and demand, Western Europe and the Far East will be the major




exporters of ethylene glycol into the U.S. through 1980 (Taylor, 1976).




There is no cushion in ethylene oxide capacity in the U.S., so imports




may be needed to relieve some tightness in the ethylene glycol market.




Worldwide capacity of ethylene glycol is expected to grow faster than




demand until 1980, making export markets very competitive.  Antifreeze




sales are expected to lag worldwide but use of ethylene glycol in




polyester fiber will increase (Taylor, 1976).




     The U.S. Bureau of the Census does not list import figures sepa-




rately for propylene glycol and butylene glycol (isomers are not speci-




fied) .  As shown in Table 13, imports of both propylene glycol and




butylene glycol into, the U.S. during 1976 exceeded 23.5 million pounds.




The major sources were West Germany (53.1%), France (19.5%) and Japan
                                     27

-------
                                                          Table  12
00
Imports

total quantity (Ibs)
country of origin '(%)
Belgium
Brazil
Canada
Finland
France
Italy
Japan
Netherlands
Swe den
Switzerland
United Kingdom
of Ethylene Glycol into the United States (U.S. Bureau
of the Census, 1972-1976)
1976 1975
45,339,410 371,225

11.2 32.5
—
0.8
_ _
0.3
— 	
36.2 30.2
34.7 23.1
__ 	 	
0.1
2.4 4.8
1974 1973 1972
,119 132,088,298 23,448,848 92,043

17.5 9.5
0.7
3.1 11.2
1.7
2.7 0.1
1.1
14.1 0.1 44.6
30.4 17.7
0.2
—
15.1 9.4 50.3

-------
                                        Table  12  (Continued)
1976 1975 1974 1973 1972
West Germany
other countries
14.2 8.9 15.1 52.0 3.4
0.5 0.1
          aCommodity number  4.,283,400,
to

-------
                                                  Table 13
                         Imports of Propylene  Glycol and Butylene Glycol into the United States'
                                        (U.S.  Bureau of the Census 1972-1976)
                                    1976
                                              1975
                               1974
                1973
              1972
CO
o
total quantity (Ibs)

country of origin (%)
  Belgium

  Brazil

  Canada

  Finland

  France

  Italy

  Japan

  Netherlands

  Sweden

  Switzerland
                                 23,663,699     17,304,595
 6.8



19.5



16.3

 3.4
                                                  25.8



                                                  43.1

                                                   1.7
                            22,831,220
 3.8



14.1



38.9

37.0
              7,152,068     1,722,487
 1.3



 0.6



13.2

76.9
                                                                                                 1.1
44.9
United Kingdom
W. Germany
other countries
—
53.1
0.9
0.3
28.9
0.2
4.5
—
1.7
0.6
7.4 54.0
—
         Commodity Number 4,283,000.  Source:  U.S. Bureau of the Census 1972-1976.

-------
(16.3%).  Imports have increased considerably since 1972,  when  the




total quantity was 1.7 billion pounds.








     5.  Market Price





     Table 14 lists the current market price of the glycols available




from representative domestic producers and distributors.   Bulk  price




refers to tank truck quantities while drum price refers to 55 gallon




drums.  The quantities quoted by Aldrich (1978) are considered  re-




search amounts.




     Table 8 lists the price trend during the last decade  for ethylene




glycol and 1,2-propanediol.  In 1968 the price of ethylene glycol




was $0.07/lb; the price remained near this level until 1974, when it




increased to $Q.14/lb.  In 1977, the average price was $0.19/lb.




The price of 1,2-propanediol increased from $0.08-0.09/lb  during




1968-1973 to $0.21/lb in 1974; in 1977, the average price  was $0.25/lb.









B.  Use




     1.  Ethylene Glycol




          a.  Current Demand





     End uses of ethylene glycol include the following:  antifreeze




(45%) , polyester fiber (35%), exports (8%), alkyd and polyester resins




(4%), latex paints and other emulsions (1%), as well as a variety of




miscellanedous uses (7%) (MCP,1977).




     The low volatility and low molecular weight of ethylene glycol




as well as its low solvent action on  automobile  finishes accounts for




its major use as a base for all-winter automobile  anti-freeze and
                                      31

-------
                                 Table  14
             Market Price  of  the  Glycols in  the United States
      Product
  Quotation (dollars)
Source
ethylene glycol



1,2-propanediol

  industrial grade



  U.S.P. grade



1,3-p ropane dio1

1,4-butanediol

  regular



  anhydrous



1,2-butanediol

1,3-butanediol
2,3-butanediol
$0.245 per pound,  bulk     Dow3 1978c

 0.305 per pound,  drums

                           Dow,  1978c

 0.265 per pound,  bulk

 0.325 per pound,  drums

 0.03 per pound, bulk

 0.36 per pound, drums

 7.50 per 100 g            Aldrich,  1978

                           GAF,  1977

 0.55-0.58 per pound, bulk

 0.62-0.67 per pound, drums

 0.97 per pound, bulk

 1.01-1.06 per pound, drums

 10.00 per 250 g           Aldrich,  1978

 0.43 per pound, bulk      Celanese, 1978

 0.495 per pound,  drum     Celanese, 1978

 14.40 per 3 kg            Aldrich,  1978

 7.45 per 100 g            Aldrich,  1978
                                     32

-------
coolant.     Related uses involve application in aircraft and runway

defrosting and de-icing formulations.  Producers of ethylene glycol

are the major suppliers of antifreeze (Kallgren, 1963).

     Use of ethylene glycol in polyester fiber is expanding, and cur-

rently accounts for about one-third of demand.  Ethylene glycol is

used as a raw material by all polyester fiber manufacturers in the

U.S. except for Tennessee Eastman; this company uses 1,4-cyclohexane-

dimethanol to produce Kodel II.  The first step in polyester manu-

facture involves the reaction of dimethyl terephthalate  with a slight

excess of ethylene glycol over a temperature range of 150-210°C in

the presence of a catalyst (Shumeyko and Mansfield, 1975).   Trade

names and producers of polyester are listed in Table 15.

     About 4% of ethylene glycol demand is for use in alkyd and poly-

ester resins.  These resins are thermosetting plastics formed by ad-

dition polymerization reactions; the resins are cured, set or hardened

into a permanent shape.  Numerous raw materials, including ethylene

glycol, are used to produce alkyd and polyester resins.   A typical

reaction to produce a polyester resin is shown below (Harper, 1975):
                           0       0
     1.  OH-CH2CH2-OH + HO-C-CH=CH-fc-OH -»

         ethylene       maleic acid
         glycol
                           0       0
                           I        I!
         H20 + HO[CH2CH2-0-C-CH=CH-C-0-CH2CH2] OH
                      c                       ^

                  ethylene glycol maleate
                  polyester

     2.  The polyester polymer units then react with styrene monomer
         in the presence of catalyst and/or heat to obtain styrene-
         polyester copolymer resin, a cured polyester.
                                     33

-------
                      Table 15
        Trade Names and Producers of Polyester Fiber
          (Shumeyko and Mansfield, 1975a)
Trade Name
          Comp any
Anavor

An gel rest

Avlin

Dacron

Esterweld

Fortrel, Fortrel 7

Fybrite

Golden Touch

Kodel

Quintess

Serene

Spectran

Strialine

Textura

Trevira
Dow Badische Co.

Fiber Industries Inc.

FMC Corp.

duPont

American Cyanamid

Fiber Industries, Inc.

Celanese Fibers Marketing Co

American Enka Co.

Eastman Kodak Co.

Phillips Fibers Corp.

Fiber Industries, Inc.

Monsanto Textiles Co.

American Enka Co.

Rohm and Haas Co.

Hoechst Fibers, Inc.
     All but Kodel manufactured using ethylene glycol
as a raw material.
                           34

-------
Alkyds are formulated from polyester-type resins.  Typical uses of


these resins are in moldings, laminated or reinforced structures,  and

coatings (Harper, 1975) .


     Exports in 1977 accounted for 8% of total demand and exceeded


253 million pounds.  They went primarily to the Netherlands (33.1%),


Taiwan (22.1%), and Belgium  (10.3%).  Export trends during the past


six years are shown in Table 16.


     Miscellaneous uses of ethylene glycol include applications in:


latex paints and other emulsions to prevent freezing; brake and shock


absorber fluids to counteract swelling of rubber and to help dissolve


inhibitors; fire retardant lubricants as a coupler; solvent for dyes


and colorants; textile  fibers, paper, leather, adhesives, and glue as


a humectant; electrolytic capacitors as a suspending medium and sol-


vent; and in the production  of ethylene glycol dinitrate, a component


in low temperature dynamites (.MCP, 1977; Dow, 1978a) .




          b.  Market Trends


     During the period 1964-1974, ethylene glycol showed an annual


growth rate of 6%  (Anon, 1976b).  Growth in the 1970's has been modest,


averaging less than 2% per year.  Future growth has been forecasted


at 3% annually through the early 1980's (MCP, 1977).


     Demand for antifreeze-grade ethylene glycol was estimated to

                       ?
be between 195-215 million gallons during 1977, up from 150 million


gallons in 1976.  In 1975, demand was about 200-218 million gallons,


reflecting hoarding by distributors and consumers due to an antici-


pated antifreeze shortage (Anon, 1977h; Anon, 1977d) .  Antifreeze
                                     35

-------
Table 16
Exports of U.S. Ethylene Glycol3 (U.S. Bureau of the Census 1972-1977)
1977 1976 1975 1974 1973 1972
Total quantity
(Ibs)
Where Exported
Canada
Mexi co
Colombia
Venezuala
Peru
Chile
Brazil
Argentina
Netherlands
Belgium
Spain
253,032,950 253,565,853 97,166,231 163,407,637 174,001,552 232,942,355
«
2.4 6.0 35.2 16.9 9.3 7.5
0.8 0.5 2.5 19.7 3.4
0.3 2.6 10.1 4.4 5.6 2.7
3.5 3.3 6.1 3.0 2.7 1.8
0.5 1.8 __ __
0.4
7.7 8.9 1.6 4.9 15.5 11.3
4.1 — 9.6 8.4 9.1 8.3
33.1 51.3 14.6 9.0 31.9 37.3
10.3 10.5 ^- ^ — 20.0
0.5 1.4

-------
                                     Table  16  (Continued)
1977
Prance —
Turkey
Switzerland * —
Mai ays a —
Philippines —
Korean Republic —
China, Taiwan 22.1
Japan 5.6
Other 8.7
1976 1975 1974 1973 1972
2.1
1.6
2.3 1.9
1.1
3.5
4.6 16.8 11.3 7.1 4.8
16.1 12.5
4.5 3.5 2.9 1.0
aCommodity Number 5120910.

-------
consumption is forecasted to remain at about 200 million gallons  per




year through 1982 (MCP,  1977) .   Although there are more cars on the




road, less antifreeze is required in the increasing number of smaller




cars.




     Demand in polyester fiber is the fastest growing end use of




ethylene glycol.  Forecasted growth in polyester demand is about 8%




per year.  However, this figure is about 50-100% less than the annual




polyester demand increase during the early 1970's (Anon, 1977d).








     2.  Propylene Glycols




          a.  Current Demand





     The major use of 1,2-propanediol (45%) is in polyester resins.




Other uses include exports (12%), pet food (12%), food and pharmaceu-




ticals  (11%), cellophane (.7%), tobacco humectant (7%) as well as




miscellaneous uses (6%)  (Anon, 1977f) .




     In polyester resins, 1,2-propanediol is esterified with diabasic




acids such as maleic and fumaric acids.  These resins are used, for ex-




ample,  in sports equipment, appliances, furniture, boat hulls, luggage,




aircraft parts, and automobile bodies (MCP, 1976).




     About 23% of 1,2-propanediol production goes into pet food, human




food, and Pharmaceuticals, in which this diol functions as a solvent,




humectant and preservative.  For example, U.S.P. grade 1,2-propanediol




is used as a solvent for flavoring materials, extract preparations, and




food colors (Dow, 1978b).  It is a humectant for baked goods and pack-




aged goods, such as shredded coconut.  1,2-Propanediol goes into products




coming in contact with foods, such as cork seals, bottle-cap linings,




and cellulose coatings for sausage and cheese.
                                      38

-------
It is also used as an intermediate for Pharmaceuticals and cosme-




tics, as a vehicle for drugs and as a solvent for medicinal chemicals,




vitamins, dyes, and perfumes (Dow, 1978b) .




     Seven percent of the 1,2-propanediol produced is used to soften




cellophane.  This market used to be the second largest consumer of pro-




pylene glycol, but use of cellophane has declined (MCP, 1976).   About 7%




is used as a humectant for tobacco formulations.




     Exports comprise 12% of 1,2-propanediol production.  Total exports




exceeded 36 million pounds during 1977, going primarily to Venezuela




(14%), Argentina  0-2.3%), the Netherlands (12.4%), and Japan (10.6%).




Exports during 1977 were at the lowest level in recent years; for example,




exports totalled  64.1 billion pounds in 1976 and 107.7 billion pounds




in 1972 (Table 17).




     Miscellaneous applications include use in:  polymeric plasticizers




Cin  vinyl compounding); and brake and hydraulic fluids as a solvent, lu-




bricant  and coupling agent (MCP, 1976).  1,2-Propanediol is also formu-




lated into some antifreeze and de-icing solutions. Because of the lower




toxicity of 1,2-propanediol compared to ethylene1 glycol, it is often




used in antifreeze which may accidently come into contact with food  (such




as in the refrigeration systems in breweries, dairies, food plants,




and  packing houses)  (Budke and Banerjee, 1971).







          b.  Market Trends





     Growth, of 1,2-propanediol production volume was 5.6% annually from




1967-1976.  A growth rate of 7-8% per year has been forecasted through




1981 (Anon, 1977f).  The major growth area for 1,2-propanediol is in




polyester resins.  These resins are expected to grow at about 12-15%
                                     39

-------
Table 17
Exports of U.S. 1,2-Propanediol (U.S. Bureau of the Census, 1972-1977)
1977 1976 1975 1974 1973 1972
Total quanti-
ty (Ibs)
Where exported
Canada
Costa Rica
Mexi co
Equador
Colombia
Panama
Venezuela
Peru
Chile
Brazil
Argentina
Netherlands
36,480,073 64,147,544 38,619,686 67,725,527 58,614,441 107,765,835
-
0.7 — 1.3 5.7 8.8 2.7
0.8
0.6 — 10.8 4.4
0.8 1.4 1.2
7.8 1.2 3.3 2.2 3.0 1.6
2.2
14.0 5.5 8.9 4.4 4.8 1.6
0.7 — 1.6
1.3
8.7 31.7 25.5 13.3 17.2 9.3
12.3 12.2 4.2 7.0 5.5 62.0
12.4 10.8 — 11.9 13.1 51.9

-------
                                    Table 17 (Continued)
1977
Belgium —
Singapore 3.2
Spain
Italy
Australia 7.3
New Zealand 5 .6
Switzerland
HongKong —
Korean Republic —
Republic of 8.4
S. Africa
China Taiwan
Philp Rep
W. Germany —
Japan 10.6
Poland
o the rs 4.8
1976 1975 1974 1973 1972
— — —
5.8 8.8
1.1
—
5.0 2.8 6.4 4.3
4.7 2.3 2.3 1.2

2.3
3.5
2.2 7.3 — 4.2
1.2
2.3 1.3
1.7
9.8 16.9 24 36.0 14.6
1.7
6.2 8.0 10.5 5.6 6.7
Commodity Number 5120912.   Source:   U.S.  Bureau  of  the  Census 1972-1977.

-------
per year (Anon, 1977f) ,  but growth here is particularly sensitive to




economic conditions in the construction, automotive, and recreational




sectors (MCP, 1976).  Consumption in other end uses will probably remain




steady, with increases in humectants offset by decreases in cellophane




and specialty antifreeze (MCP, 1976).







     3.  Butylene Glycols





     1,3-Butanediol is used primarily as an intermediate in the manu-




facture of polyester plasticizers, particularly in polyvinyl chloride




but also in synthetic rubbers, nitrocellulose, polyvinyl acetate, ethyl-




cellulose, and polyvinyl butyral resins.  1,3-Butanediol is also used




as a gelling agent  for gelatin, proteins, and cellulose nitrate.  High




purity 1,3-butanediol (99.00%) serves as a solvent for several natural




and synthetic  flavoring substances such as spearmint, peppermint, and




orange oils  (Wagner, 1966).  It functions in some hand cleansers, lotions,




and other cosmetic  formulations (Budke and Banerjee, 1971).  Pyroborate




and boronate esters of 1,3-butanediol are used in jet-fuel to improve




high temperature stability; these esters also inhibit the growth of mi-




croorganisms in jet  fuels.  The sulfite ester  .can be used to stabilize




cellulosic esters  (Wagner, 1966) .




     The major market for  1,4-butanediol in the U.S. is in the prepara-




tion of tetrahydrofuran  (57%).  Tetrahydrofuran is combined with C02




to form adipic acid and  is also used as an intermediate for other chemi-




cals.  Other applications  include use in acetylenic chemicals (25%),




polyurethanes  (9%), polybutylene  terephthalate (PBT, 7%), as well as




miscellaneous  markets (1%).  Current domestic demand for 1,4-butanediol




is 175 million pounds (Brownstein and List, 1977) .  DuPont predicts
                                     42

-------
that demand for 1,4-butanediol will increase rapidly over the next few




years for use in engineering plastics and polyurethanes.  DuPont employs




1,4-butanediol capitively in the manufacture of tetrahydrofuran and,




since 1975, also in polybutylene terephthalate resin (trade name "Rynite")




(Anon, 1976a).  BASF uses 1,4-butanediol to produce polybutylene-tere-




phthalate and to make polyurethanes injection molded thermoplastics and




shoes (Anon, 1977i).




     2,3-Butanediol is used as a solvent for dyes, as a humectant and as a




coupling agent  (Budke and Banerjee, 1971).  1,2-Butanediol is used in




the production of polymeric plasticizers (Hatch and -Matar, 1978b).









C.  Possible Alternatives to Use




     A major use of ethylene glycol is in polyester fiber.  To retain the




special qualities of polyester, a suitable substitute might not exist.  In all




polyester formulations,  ethylene glycol is used as the raw material, with




the exception of Kodel II which uses 1,4-cyclohexanedimethanol.  Kodel II




has a somewhat higher melting point, and has bulkier and deeper-dyeing fibers




than polyester made from ethylene glycol (Shuiaeyko and Mansfield, 1975).




     Other raw materials besides 1,2-propanediol and ethylene glycol




are currently used to produce alkyd and polyester resins (Harper, 1975).




These include a variety  of organic alcohols, such as neopentyl glycol,




which could be used in place of ethylene and propylene glycols.  Use of




1,2-propanediol in food and pharmaceuticals would be a difficult market




for which to find suitable substitutes.  Other polyols, such as glycerine,




sorbitcl, and mannitol,  which are also used as food additives have some-
                                     43

-------
what overlapping properties as 1,2-propanediol and might substitute for




it.  Use of 1,2-propanediol as a solvent in Pharmaceuticals would be




particularly difficult to substitute because of its relatively low toxi-




city.









D.  Entry into the Environment




     Only limited monitoring data on the glycols are available.  These




are reviewed in the following section on sources of entry into the en-




vironment .





     1.  From Production




     Ethylene glycol and 1,2-propanediol have been identified as com-




ponents of the wastewater from production facilities (Zeitoun and




Mcllhenny, 1971).  In 51 samples at two ethylene glycol plants, the




concentration of ethylene glycol in the wastewater ranged from 680-2,300




ppm and averaged between 1,003-1,306 ppm.  In a similar number of samples




at two 1,2-propanediol plants, the level of this glycol ranged from




355-2,550 ppm and averaged between 960-1,140 ppm.  Other components of




the wastestreams are described in the section on waste handling.  As




discussed in the section on waste handling, these wastestreams can be




handled by biological treatment, in which case the glycols would not be




entering the environment.




     A source of air pollution during 1,2-propanediol manufacture is the




venting of excess vapor from the evaporation and drying towers, as shown




in Figure 2 (EPA, 1975).  It is not known whether 1,2-propanediol is a




component of this vented vapor.
                                    44

-------
     Other minor sources of environmental contamination might be from




fugitive emissions at valves, seals, and vents (the glycols would be




volatile, however, only at elevated temperatures) and from accidental




leaks and spills.




     The potential for environmental contamination will certainly de-




pend on the design of a given production facility and to what extent




spent glycol is recycled (see section on waste handling) and treatment




of waste streams is practiced.  As such data are unavailable, it is




difficult to accurately estimate the environmental hazards from produc-




tion facilities in general.







     2.  From End Product Manufacture




     Ethylene glycol was identified as a constituent of the wastewater




of a polyester fiber plant in Poland.  The level of ethylene glycol




ranged from 200-440 mg/& and averaged 220 mg/£ (Grabinska-Loniewska,




1974a).







     3.  From Use and Disposal




     The major source of environmental contamination by ethylene glycol




and 1,2-propanediol is likely from the disposal of spent antifreeze,




but no estimates are available assessing the magnitude of this problem.




The upper limit of this contamination is based on antifreeze use and the




assumption that all is discarded to the environment.
                                    45

-------
     Another source of environmental contamination by  glycols  is  runoff



of de-icing fluids.  Aircraft de-icing fluids,  for example,  are sprayed



on airplanes and the runoff can enter the airport sewer collection sys-


tem or contaminate surface waters.   The composition of de-icers avail-


able from two manufacturers contained 16 and 38% 1,2-propanediol  and


32 and 56% ethylene glycol; other characteristics of de-icers  appear


in Table 18 (Jank et al.,  1974).
                                 Table 18

                Composition of De-icers (Jank et al.,  1974)

Composition (%)
water
ethylene glycol
propylene glycol
Dow

50
32
16
UCAR

5
56
38
                  (1,2-p ropanediol)



                  inhibitor package        2         1


                Characteristics



                  TOCa (mg/41        261,000    460,000


                  BODb (mg/£)        362,000    712,000


                  pE                    8.0         8.7
                    a.
                    TOC  =  total  organic  carbon.


                    BOD  =  biological  oxygen  demand.
                                     46

-------
     Measurements at the Montreal International Airport revealed that de-




-icers  are used about 150 days of the year.  Drainage is into two manholes




 (storm sewer); during February-March 1972,  the glycol content in the runoff



ranged from 0-4,780 mg/A .  The BOD of the waste ranged from 200,000-




400,000 mg/A  (Schulz and Comerton, 1974).  Jank et al. (1974)  studied




the biological treatment of de-icing runoff on a laboratory scale.   An




activated sludge system treating de-icing fluid and sewage at  less  than




10°C resulted in an effluent having a BOD of less than 20 mg/A and  TSS*




of less than  25 mg/5,.  The authors caution that growth of filamentous




microorganisms might cause a problem in full-scale plants.







     4.  From Transportation





     Transportation accidents are another possible source of environmental




contamination.  Estimates of the safety of ethylene glycol transport by




barge, rail,  and truck were provided by Arthur D. Little, Inc. in 1974.




Estimates were based on one selected route:  the shipment of ethylene




glycol from Belle, West Va. to Pittsburgh, Pa., covering 335 miles  by




barge, 290 miles by rail, or 200 miles by truck.  According to the  U.S.




Army Corps of Engineers, the quantity of ethylene glycol shipped annually




between this  origin and destination exceeds 100,000 tons.  Based on past




accidents, the number of trips made by each mode of transport and other




factors, the  following estimates were made of the probable annual number




of accidents  causing release of cargo:  0.0018 by barge; 0.014 by truck;




and 0.055 hy  rail.  These figures represent the hypothetical  situation




where  the entire commodity would be transported by each mode.  Arthur




D. Little, Inc. also took this model further and estimated the spill




pool radius,  the hazardous radius, population exposure, probability




*total suspended solids





                                      47

-------
of ignition, annual property damage and the recurrence interval.   These

estimates are listed in Table 19.   Based on these estimates,  rail

transport will occur more frequently,  expose more people,  have a  greater

hazard area and will damage more property than the other two  modes of

transportation.


     5.  From Microorganisms

     2,3-Butanediol occurs in the environment as an endproduct of fermen-

tation by various strains of enteric bacteria (Enterobacteriaceae) such

as Aerohacter, Klebsiella, and Serratia (Brock,  1970) .  Typical end pro-

ducts of fermentation by Aerobacter are:

                                    moles product/100  moles
                                    glucose fermented
                2,3-Butanediol            66.5

                C02                      172

                K2                        36

                formic acid               18

                acetic acid                0.5

                lactic acid                3

                ethyl alcohol             70


     Serratia marcescens produces 2,3-butanediol from agricultural waste

materials (Bahadur and Dube, 1959) .   The acid hydrolyzates from wheat
                      c
straw, wheat bran, rice straw, and rice polishings gave yields of 31-40%

butanediol on the basis of sugar consumed.

     An actively growing culture of  Bacillus polymyxa produced 2,3-bu-

tanediol, but the quantity decreased over a 60-day period (Bahadur and
                                     48

-------
                                 Table 19
                Hazards of Ethylene Glycol Transportation'
                        (A. D. Little, Inc., 1974)

spill pool radius (feet)
hazard radius (feet)
hazard area (acres)
relative exposure (%)
Barge
NA
NA
.04b
16/84
Truck
56
56
.22
16/84
Rail
104
104
.78
6/94
  urban/rural

expected number of annual spills     .0018     .014           .055

expected annual number of people     0         .001/.0002     .006/.004
  exposed urban/rural0

expected property damage $c          0         20/4           103/60
  urban/rural

recurrence interval                  555       71.4           18.2
  (years)
     Calculations are based on the assumption that each mode handles
100% of the quantity shipped.

     For spills into water which do not ignite the water toxicity hazard
distance (feet) measured downstream from spill location for a 500 feet
wide, 10 feet deep river flowing at 2.3 feet per second.  Assumes verti-
cal dispersion rate at 1.0 feet per minute until uniform mixing is achieved.
    c
     Although ethylene glycol is not considered flammable or capable  of
chemical decomposition, its potential to cause hazards is assessed in
terms of number of people exposed and dollar value of exposed property.
It is, however, recognized that this approach is conservative.  In any
event, it provides a bcasis for comparing relative damages associated
with transportation by alternative modes.

     Average number of years between accidents.
                                    49

-------
Ranganayaki, 1961) .   Several strains of lactobacilli and staphylococci

produce 2,3-butanediol,  which is then further metabolized (Cantoni et

al., 1965) .

     Filamentous fungi,  such as Rhizopus nigricans and Penicillium ex-

pansum, also produce 2,3-butanediol (Fields and Richmond, 1967).  In a

laboratory experiment, the amount of 2,3-butanediol identified after

5-14 days incubation of R.. nigricans on an artificial or apple juice

medium ranged from 43.0-230.0 yg/ml.  For P_. expansum levels ranged from

104.0-343.0
     6 .  From Other Sources


     Ethylene glycol and ethylene chlorohydrin are degradation products

of ethylene glycol; these compounds are found as residues after sterili-

zation and fumigation of various organic materials and synthetic objects

with ethylene oxide (Gardner, 1978) .

     Ethylene glycol, 2,3-butanediol and 1,2-propanediol were identified

as three of 479 water-soluble components of cigarette smoke (Schumacher

et al., 1977) .




E.  Waste Handling

     1.  Characteristics of Waste Stream


     Characterization of waste streams from ethylene glycol manufacture
                      f
appears in Table 20.  Data for flow, biological oxygen demand (BOD) and

chemical oxygen demand  (COD) were submitted by ten manufacturers to EPA

during 1975 or earlier.  These values are quite disparate; flow ranged

from 9.4-14,300 gal/1,000 Ibs , BOD from 0.34-27.7 lb/1,000 Ibs and COD

from 0.80-209.0 lb/1,000 Ibs.  Data from three manufacturers (Dow,
                                      50

-------
                                Table  20
     Flow,  Biological  Oxygen  Demand  (BOD) and Chemical Oxygen Demand
          (COD)  of Waste  Streams Generated at Various Plants Dur-
              ing Ethylene Glycol Manufacture (Train,  1975)a
Source
Dow Chemical Co .
Freeport, Tex.6
Union Carbide Corp.
Ponce, P.R.
Jefferson Chemical Co .
Port Nech.es, Tex.
Union Carbide Corp .
Taft, La.
Shell Chemical Co.
Houston, Tex.
Olin Corp.
Brandenburg , Ky .
Texas Eastman Co .
Longview, Tex.
Union Carbide
Port Lavaca, Tex.
Houston Chemical Co .
Houston, Tex.k
Citgo Corp .
Lake Charles, La.
Flow
gal/1,000 Ib
584
29
178
66
5,430
7,000
9.4

272

14,300
128
BOD
lb/1,000 Ib
0.34
0.5
1.02
3.5
10.5
12.4
27.7

7,6

11.3
0.07
COD
lb/1,000 Ib
8.76
0.80
2.18
9.7
22.6
209.0
48.6

10.4

12.2
11.3
    aData received by EPA from the individual manufacturers;  data were
generally collected over a 30-day period or longer.

     Combined load for ethylene glycol and ethylene  oxide  production.
    £
     According to Citgo, these data are suspect due  to  possible  incor-
rect analysis.

                                                     MATERIAL BELONGS TO:1
                                                     US EPA TOXICS LIBRARY
                                                       401 MSTSVV/TS-793
                                                     WASHINGTON, DC 2C-4nO
                                                          (202) 260-3944
                                     51

-------
Union Carbide, and Houston Chemical)  are for combined process wastes

from both ethylene glycol and ethylene oxide production.  According to

EPA,. (1974). , a source of waste streams is the condensate from the dehy-

drator, which is only partly recycled (refer to Figure 1) .  Additional

measurements of waste water generated during ethylene glycol manufacture

at two plants were made by Zeitoun and Mcllhenny (1971) of the Dow Chemi-

cal Company, as shown below:
                                             Average values
                CODa, ppm                    1,630-1,850

                TODb, ppm                    1,730-1,920

                NaCl, %                      10.72-12.90

                total dissolved solids, %    11.12-13.30

                ethylene glycol, ppm         1,003-1,306

                ethylene oxide, ppm              60

                ethylene dichloride, ppm         16

                ethylene chlorohydrin            15

                dichloroethyl ether              29
                    Q
                     Chemical oxygen demand.


                     Total oxygen demand.
     In 1,2-propanediol manufacture, waste waters are generated, for ex-

ample, during the evaporat ion-drying operation and the distillation pro-

cesses (refer to Figure 2) (EPA, 1975) .  Measurement of waste water at

one plant yielded the following:
                                     52

-------
     flow              660 gal/1,000 Ib

     BOD5              0.016 lb/1,000 Ib

     COD5              0.055 lb/1,000 Ib

     total organic     0.006 lb/1,000 Ib
       carbon

Zeitoun and Mcllhenny (.1971) of Dow Chemical Co. provided additional data

on waste water generated during 1,2-propanediol manufacture at two plants

(one a pilot plant) tabulated below:
                                            Average values
                COD, ppm                     2,138-3,086

                TOD, ppm                     2,198-4,398

                % NaCl                        9.8-10.3

                total dissolved solids       10.1-10.65

                propylene glycol              960-1,140
                (1,2-propanediol)

                propylene oxide                28-196

                epichlorohydrin                25-138

                propylene chlorohydrin         42
     2.  Treatment o_f Waste Stream


     Measurements by Zeitoun and Mcllhenny (1971) reveal that at both

ethylene glycol and 1,2-propanediol production facilities, the waste-

water is characterized by a high salt content  (10-13%), excess alkalinity

(0.05-0.45%, as sodium hydroxide) and the presence of  several organic

compounds.  Zeitoun and Mcllhenny tested several alternative treatment
                                      53

-------
methods to handle these wastes, which are briefly described below.








     i.  Solvent Extraction — Solvent extraction of glycol wastewater




with secondary or tertiary amines produced:   a)  a raffinate that was




low in glycol and was salt saturated (this could be recycled for use  in




hydrolysis) and b) a product enriched with glycol and almost free of




salt Cglycol would be recovered). .  The authors found this system to be




uneconomical.







     ii.  Carbon Adsorption — Adsorption on activated carbon was not




feasible because of the low capacity of activated carbons for glycols.







     iii.  Membrane Separation — Cellulose acetate membranes did not




efficiently separate the salt and the glycol.







     iv.  Biological Treatment — This was the preferred method of treat-




ment.  Organisms from natural saline environments were screened for their




capacity to utilize glycols.  The biodegradability of glycols is discussed




in the next section on fate and persistence.  Biological oxidation of




propylene glycol wastewater in batch and continuous laboratory units




gave a total oxygen demand (TOD) removal efficiency of 86-88% at a resi-




dence  time of 12 hours.  An activated sludge pilot plant was built which




removed more than 90% of the TOD at a residence time of 8.0 hours.  Design




parameters for this pilot plant are presented by Zeitoun and Mcllhenny




(1971) .




     Grabinska-Loniewska (1974a) found that removal of ethylene glycol




(.0.11-0.50 mg/mg/day) wastes generated during polyester fiber production




was 8Q-1QQ% complete using activated sludge.  The percent removal depended
                                      54

-------
on the initial sludge load; efficiency decreased with increasing sludge




load (100-500 ml/g total sludge volume tested).




     Japanese investigators Kawami and Torikai (1976). described the




electrolytic treatment of a synthetic effluent (COD = 20,000-30,000 ppm)




containing 57, ethylene glycol.  The effluent was electrolyzed using a




platinum or lead dioxide anode.  Products of decomposition were CO and Q£




when using the platinum anode and C02 when using the lead dioxide anode.




The decrease in COD was 73 and 90%, respectively, for the two anodes.







     3.  Diol Recovery





     Cox and Wilkes (1975) in a patent assigned to PPG Industries, Inc.




described a method for recovering ethylene glycol used in the manufacture




of polyesters.  In one step of this manufacturing process, ethylene gly-




col is esterified with dimethyl terephthalate or terephthalic acid.




Depending on the nature of the process used, the residual glycol stream




(or "spent glycol") can be contaminated with ethylene glycol, dimethyl




terephthalate, salts of terephthalic acid, methanol, water, acetals,




higher glycols, as well as other solid and liquid contaminants.  The




glycol stream may contain 40-90 weight percent ethylene glycol, so the




recovery of this glycol plays an important role in the economics of




polyester manufacture.  Recovery involves subjecting the spent glycol




to a sequential flash evaporation-distillation or distillation-flash




evaporation-distillation procedure.  Dimethyl terephthalate impurities




are rendered non-volatile by the addition of alkali metal hydroxide




prior to flash evaporation.  Ethylene glycol that is recovered from




spent glycol can be used in antifreeze compositions or, if pure enough,




it can be reused in the polyester manufacturing processes.





                                       55

-------
     Mattia (1971)  in a patent assigned to Day and Zimmermann,  Inc.  de-




veloped a process for recovering organic compounds present in aqueous




streams at levels between 50-5,000 ppm.  He suggested this method is




applicable for the recovery of ethylene glycol from polyester plant




effluent streams.  The recovery process utilizes a system of absorbers




to strip organic compounds from the aqueous stream.  Steam desorption




is accomplished by a thermocompressor, which recycles regenerating




steam through the adsorbers.









F.  Fate and Persistence in the Environment




     1.  Biological Degradation





     The glycols are capable of being degraded by a variety of bacterial




strains.  Several investigators identified those strains which can utilize




the glycols as sole carbon sources. Other investigators studied the rate




of biodegradation under controlled laboratory conditions.  Both types




of studies are discussed for each glycol.







          a.  Ethylene Glycol




               1)  Microbial Metabolism





     Grabinska-Loniewska (1974a) isolated strains of bacteria capable




of decomposing ethylene glycol from activated sludge used to treat poly-




ester fiber plant wastes.  Among other constituents, these wastes con-




tained ethylene  glycol (200-440 mg/Jl) , methanol (215-450 mg/fc) , and




formaldehyde (.60-68 mg/Jl) .  Of 44 strains of bacteria isolated from




activated sludge which degraded the three compounds, Pseudomonas bac-




teria were most prevalent. Less numerous were Achromobacter, Flavobac-




terium, Mycobacterium, and Xanthomonas.  In another study, Grabinska-
                                     56

-------
Loniewska (1974b) examined each of the 44 strains as  sole carbon  sources




for various compounds, ranking growth as excellent,  good, fair, or poor.




The following summarizes data obtained for ethylene  glycol:
Species of
Organism
Acetobacter
Achromobacter
Alcaligenes
Arthrobacter
Flavobacterium
Micrococcus
Mycobacteri"™
Pseudomonas
Sarcina
Xanthomonas

No.
Strains
1
4
1
1
2
1
3
26
1
4
Growth on ethylene glycol
excellent good fair poor No
1
3 1
1
1
1 1
1
3
10 7414
1
12 11
Excellent or good growth was obtained with most (70%) of the strains tested,




     Gonzalez et al. (1972) isolated a bacterium (ATCC 27042) capable of




using ethylene glycol as a sole carbon source from a brine pond.  This




gram-negative nonmotile rod required a medium containing at least 0.85%




NaCl for growth but tolerated up to 12% NaCl.




     Haines and Alexander (1975) isolated a species of microorganism from




soil (Pseudomonas aeruginosa) capable of degrading ethylene glycol.  Based




on measurements of 02 consumption, ethylene glycol (added at 1.0 mg car-




bon per bottle) was degraded in two days.
                                     57

-------
     Jones and Watson (1976) studied  the  catabolism of ethylene  glycol


by a bacterium  (possibly Acinetobacter)  isolated from sewage.   Cell-free


extracts of  the  bacterium grown on ethylene glycol contained high (0.175


Umol/min/mg  protein),  ethylene glycol  dehydrogenase activity; ethanol


and 1-butanol were also oxidized.  NAD+ and NADP"1" were utilized.  In


addition, bacteria grown  on ethylene glycol showed a 40-fold  increase


of a NAD+-dependent glycolaldehyde dehydrogenase activity  and  an increase


in glycerate-pathway enzyme activity.


     Child and Willetts (.1978) described the metabolism of ethylene


glycol by a  species of Flavobacterium (NCIB 11171) .  This  bacterium,


isolated from pond water, is capable  of using ethylene glycol  as its


sole carbon  source, optimal growth was obtained at 30 mM ethylene glycol.


Several experimental procedures were  used to identify the  metabolic path-


way for ethylene glycol metabolism, including a) assaying  for  specific


metabolites  and  enzymes, b) comparison of oxidation by various substrates,


c) measuring incorporation of ^C-labeled ethylene glycol  into the


ethanol-soluble  fraction of washed suspensions.  The following pathway


was suggested by the results of these experiments.

                                   (•   CoASH <-^
           ethylene'-* glycollate -» glyoxylate^-* oxalyl-CoA-Aoxalate
           glycol
                   tartronic      ,,
                   semialdehyde   C°2
                        ;                    V
                   glycerate

                   phosphogly cerate                     x
                       c|                   CO2-
                   phosphpenofpyruvate -* pyruvate -^ acetyl-CoA



                                      ""--v oxaloacetate citrate
                                             TCA cycle   |              \
                                        tnalate

                                             ^	

                                                    "2 X CO2

              -, Catabolic pathway;	, Anaplerotic pathway. TCA, tricarboxylic acid.


                                       58

-------
               2)  Rate of Degradation


     Evans and David (1974) evaluated the biodegradation of ethylene

glycol in river water under controlled laboratory conditions.   Samples

from four types of river waters were used:  a)  major watercourse with

a constant slow flow rate; b) major tributary with a varied flow running

through agricultural land; c) fast running river originating in an up-

land stream; d) slow flowing river passing through agricultural land.

To the samples were added 2 or 10 mg/Jl ethylene glycol.  Biodegradation

was monitored over 14 days at 4, 8, or 20°C.  Remaining glycol was de-

termined by oxidation with acidified permanganate to an aldehyde and

subsequent reaction with 3-methylben2othiazolone hydrazone hydrochloride

to give chromogens measured at 630 nm.  Ethylene glycol biodegraded com-

pletely within three days in all river waters tested at 20°C.  On day

one, Q-1.8 mg/Jl  ethylene glycol remained when 2 mg/£ had been added;

1.8-9.1 mg/Jl remained in samples to which 10 mg/Jl had been added.  At

8°C, hiodegradation was complete within 14 days for each river sample;

the rate of degradation in rivers a) and b) was slow for the first seven

days but increased thereafter.  At 4°C, the rate of degradation was low;

only about 0.2 mg/Jl per day were degraded.  In most samples tested at

this temperature, partial degradation occurred by day seven but degrada-

tion was not complete by day 14.

     The theoretical oxygen demand for ethylene glycol is 1.30 mg/mg.
                      c
This value represents the oxygen needed for complete conversion to C02

and EaO.  The actual chemical oxygen demand is 1.29 mg/mg, as  calculated

by Price et al.  (1974).

     Lamb and Jenkins (1952) studied the biological oxygen demand in  a
                                      59

-------
solution of ethylene glycol over a 20 day period at  20°C.   Ethylene gly-




col (2.5 mg/£) was added to a BOD bottle with mineralized  dilution  water




and settled sewage seed.  The percent of theoretical BOD after 5, 10,




15, and 20 days was 12.5, 51.8,  71.0, and 78.0%, respectively.




     Fitter (1976) determined the biodegradability of ethylene glycol




based on measurements of chemical oxygen demand.  About 1,000-1,500




ml ethylene glycol were added to adapted activated sludge  so  that the




initial COD was 200 mg/£.  Ethylene glycol was the only source of carbon




provided.  Over 120 hours, 96.8% of the ethylene glycol had been de-




graded.  Expressed as a rate, 41.7 mg COD were degraded per gram per




hour.  According to the author,  ethylene glycol is readily decomposed.




     Bedard (1976) evaluated the biodegradation of ethylene glycol, anti-




freeze, and other organics over a 30-40 day period.   The quantitative




measure used was the Refractory Index (RI) which is  defined as a ratio




of the oxygen required for biological stabilization  in a long-term  War-




burg Respirometer (.the ultimate Biochemical Oxygen Demand  or  BOD)   to




the amount of oxygen required for complete oxidation to end products of




C02, H20, and N03~(the ultimate oxygen demand or UOD) .  Domestic raw




sewage was used as seed.  Compounds having a high R.I. value  degrade




readily and are judged safe for disposal into municipal biological  sewage




systems.  Bedard cautions, however, that a high R.I. is a  function  of




the favorable environment of the Warburg flask and similar results  may




not be obtained in the'natural environment in such a short time. Both




ethylene glycol and antifreeze had high R.I.:  0.76  and 1.12, respectively,




as shown in Table 21.
                                     60

-------
                                 Table 21
             Biodegradation of Ethylene Glycol,  Antifreeze,  and
                      1,2-Propanediol (Bedard,  1976)

ethylene glycol
antifreeze
propylene glycol
(1 , 2-p ropanediol)
Ultimate Biochemi-
cal Oxygen Demand
CBQDu)a Cmg/4)
207
301
326
220
Ultimate
Oxygen .
Demand (UOD)
(mg/Jl)
271
268
420
419
Refractory
Index (R.I. =
BOD /UOD)
0.76
1.12
0.78
0.52
      Measures quantity of Q£ used by bacteria originating from a  domes-
tic raw sewage seed as an energy and growth supply over a 30-40 day period,

      Amount of oxygen needed for complete oxidation to end products of
C02, H.20, and N03~.
     Price, Waggy, and Conway (1974) determined the biodegradability  of

ethylene glycol and other petrochemicals in freshwater.  Settled domestic

wastewater Cseed material) and aerated water were added to BOD bottles.

The test chemicals were added to yield a final concentration of 3,  7, and

10 mg/&.  Dissolved oxygen was monitored over 20 days.   The percent bio-

oxidation was calculated on day 5, 10, 15, and 20 using this equation:
                      «
                                    100(0' -OJ
                    bio-oxidized
                                     C -ThOD
                                      X
where 0'  • cumulative oxygen uptake in the sample from day zero to the
        5

            day of interest
                                      61

-------
     0,  = cumulative oxygen uptake in a blank (containing the  same  amount



          of seed material) from day zero  to  the  day  of  interest



     C  = initial concentration of the compound
      x


    ThOD  = theoretical oxygen demand per  mg  of compound for complete



            conversion of the compound to  C02 and
As shown in Table 22, the bio-oxidation for ethylene glycol  was  34%,  86%,



92%, and 100% after 5, 10, 15,  and 20 days, respectively.



     Price et al. (1974)  also determined the biodegradation  of ethylene



glycol in synthetic seawater. .The seed used was  from seawater taken  from



Lavaca Bay, Texas.  Using the formula described above,  the bio-oxidation



was 20, 60, 65, and 77% after 5,  10,  15, and 20 days,  respectively (Table



22).  That is, rate of degradation was slower in  seawater.






          b.  Propylene Glycols



               1)  Microbial Metabolism




     Gorban and Petrenko  (1972)  identified several species of  Pseudomonas



capable of using 1,2-propanediol  as a sole source of carbon.   They es-



tablished a maximum permissible  concentration of  700 mg/£  of  1,2-pro-



panediol for biological degradation to precede.



     Morihara (1965) determined  that  Pseudomonas  aeruginosa  (Strain IFO



3455) is capable of good growth  and high protinease production when 1,2-



propanediol was supplied as the  sole source of carbon;  optimal growth



was obtained in a 5% solution.



     Yagi and Yamada (1969) isolated 60 strains of 1,2-propanediol-utiliz-



ing microorganisms from 150 soil  samples.  Thirty strains were bacteria,



29 were molds and one was an actinomycete .  One strain, Arthrobacter
                                     62

-------
                                 Table 22
           Biodegradation of Ethylene Glycol and 1,2-Propanediol
                           CPrice et al., 1974)
                                   Ethylene     ,  „ „      ...
                                      '  ,       1,2-Propanedxol
      theoretical oxygen            1.30            1.68
        demand Cmg/mg).

      measured chemical oxygen      1.29            1.63
        demand (mg/mg)

      biodegradability (.% bio-
        oxidation)
Freshwater
Day




5
10
15
20
a
34
86
92
100

62
68
75
79

Salt Water"
Day



5
10
15
20
20
60
65
77
55
72
73
83
         
-------
5% 1,2-propanediol was compared to growth in nonexposed controls over 2-4




weeks.  In eleven species, growth in 1,2-propanediol and in controls was




comparable.  However, three species were intolerant to 1,2-propanediol




(Mycobacterium tuberculosis, M. bovis,  and M. kansasii) and did not grow.




      Resting cells of the yeast Hasenula miso IFO 0146 grown on ethanol




 are capable of oxidizing dl-1,2-propanediol to acetol and 1,3-propanediol




 to B-hydroxypropionic acid (Harada and Hirabayashi, 1968).  Two ml of a




 1% solution of each glycol were tested.  Oxidation products were identi-




 fied by paper chromatography-







                2)  Rate of Degradation





      1,2-Propanediol was designated as having a "high degradability"




 by Bedard (.1976) based on its  high refractory index (R.I.).  As defined




 in Table 20, the refractory index is the ratio of the ultimate biochemi-




 cal oxygen demand to the ultimate oxygen demand.  In two replicate analy-




 ses, the R.I. of 1,2-propanediol was 0.78 and 0.52.




      The theoretical oxygen demand for 1,2-propanediol is 1.68 mg/mg.




 This value is the oxygen required for complete conversion to C02 and H20.




 Price et al. (1974) measured the actual chemical oxygen demand to be




 1.63 mg/mg.




      Lamb and Jenkins (1952) followed the biological oxygen demand of




 2.5 mg/£ 1,2-propanediol added to a BOD bottle with mineralized dilution




 water and settled sewage seed.  The percent of theoretical BOD satis-




 fied after various days is as  follows:
                                      64

-------
Day
5
10
15
20
30
40
50
% theoretical BOD
2.2
59.8
61.5
66.4
64.0
72.2
68.0
     Price et al. (1974) determined the biodegradability of 1,2-propanediol




in freshwater over 20 days using the procedure described for ethylene




glycol in the previous section.  The data appear in Table 22 and show




that after 5, 10, 15, and 20 days, the percent of 1,2-propanediol bio-




oxidized was 62%, 68%, 75%, and 79%, respectively.




     Using synthetic seawater, Price et al. (1974) determined the bio-




degradafaility of 1,2-propanediol to be 55% after five days and 83% after




20 days, comparable to degradation in freshwater (Table 22).
          c.  Butylene Glycols




               1)  Microbial Metabolism






     Tsukamura (1966) showed that some strains of mycobacteria are capable




of using butanediols as sole sources of carbon.  Of 132 strains tested,




56 showed growth with 1,3-butanediol, 8 with 1,4-butanediol, and 59 with




2,3-butanediol.
                                      65

-------
     Resting cells of the yeast Hansenula miso IFO 0146 grown on ethanol




oxidized meso-2,3-butanediol,  dl-l,3-butanediol and 1,4-butanediol to




acetoin, g-hydroxybutyric acid and a-hydroxybutyric acid,  respectively




(Harada and Hirabayashi, 1968).  These products were identified using




paper chromatography.  The diols were tested using 2 ml of a 1% solu-




tion .




     2,3-Butanediol at 0.1 or 1.0 molar concentration was  not inhibitory




to the growth of Pseudomonas fragi (Pinheiro et al., 1968) .







               2)  Rate of Degradation





     1,4-Butanediol is readily degradable.  Fitter (1976)  added about




1,000-1,500 ml to activated sludge adapted for 20 days.  During 120 hours,




98.7% of the glycol had been degraded based on reduction of initial COD.




The  rate of degradation was expressed as 40.0 mg COD/g/hour.







     2.  Chemical Degradation





     The glycols are quite stable compounds.  As discussed in the previous




section, they are all readily oxidized by microorganisms,  and would be




expected to be biodegraded before chemical degradation became significant.







     3.  Transport Within and Between Media





     The subject glycols are soluble in water in all proportions, and




are  denser than water.' When spilled in a body of water, they would




sink, then dissolve.




     The glycols have a very low vapor pressure at ambient temperatures




(Tables 2-4) so evaporation from water or land to the atmosphere will
                                     66

-------
occur slowly.  Runoff of spent antifreeze or de-icing fluids can occur;




for example, the runoff of deicing fluids, which contain ethylene and/or




propylene glycol, can enter airport sewer collection systems or contami-




nate surface waters (see section II-D-3).








     4.  Persistence and Bioaccumulation





     The glycols are subject to moderately rapid breakdown by both accli-




mated and unacclimated  soil, water, and sewage microorganisms, as dis-




cussed in section II-F-1, thus precluding persistence in the environment.




There is no  evidence to suggest that the glycols would bioaccumulate.









G.   Analytical  Methods





     Various methods are  available for  the  identification and separation




of  glycols.  Chemical  and chromatographic methods used during the last




few years are  reviewed in this section. The  reader  is referred to Budke




and Eanerjee 0-971)  for a comprehensive review  of older papers.








      Colorimetry





      A number  of chemical methods have  been developed for  detecting




glycols  in  aqueous  solutions or  in biological material.  These  methods,




in  most  cases,  involve the  oxidation of the glycol  with subsequent  re-




action with a  reagent.  In  some  tests,  bromine water or potassium per-




manganate is used to oxidize ethylene glycol to glycolic aldehyde and




propylene glycol to acetol.  Adding the reaction product to Fehlings




solution,  ammoniacal silver nitrate or Nessler's reagent will produce




a positive  test.  With phenolic reagents (such as resorcinol, thymol,
                                      67

-------
and codeine) in sulfuric acid,  characteristic color for ethylene and pro-




pylene glycol are obtained (Budke and Banerjee,  1971) .




     Ethylene and propylene glycols are oxidized by periodates to al-




dehydes which can be quantitated colorimetrically.   For example, Rajogopal




and Ramakrisnan (1975) used sodium metaperiodate (NalO^)  to oxidize




ethylene glycol to formaldehyde for determinations  of ethylene glycol




in blood and tissue.  Excess periodate was  reduced  by arsenite after




which chromotropic acid was added;  the purple color was  developed in a




boiling water bath, and measured at 550 nm.  From 98-100% of known amounts




added to blood and liver homogenate were measured by this method.  Russell,




McChesney and Golberg (1969) also oxidized  ethylene glycol with periodate




for determinations in biological material.   A protein-  and carbohydrate-free




filtrate was prepared^then oxidized .with periodate.  The  formaldehyde which




was produced is converted to a  dihydrolutidine derivative.  The ahsorbance




of the solution was read in a spectrophotometer at  412  my.  The precision




of this method was 3-4% in tests with known amounts of  ethylene glycol in




urine, eerum, and tissue.  In this method,  metabolites  of ethylene glycol do




not interfere.




     Another method using oxidizing action  of ethylene  glycol with perio-




date was described by Sunshine  (1969)  for determinations  in blood or




urine.  The oxidation product was condensed with Schiff's reagent to give




a characteristic colored product whose absorbance is read at 555 nm.  Re-




covery was 100 ±5%.    «




     Efstathiou and Hadjiioannou (1975) developed a method for determin---




ing the rate at which ethylene  glycol, 1,2-propanediol, or 2,3-butanediol




is oxidized with periodate.  The reaction rate is followed with a per-




chlorate ion selective electrode; the time  is measured  for the reaction
                                       68

-------
to consume a fixed amount of periodate . (and therefore for the potential




to increase) which is related to the glycol concentration.  Measurements




were made over 3-18 mg/28 ml of glycol with a relative error of 0.7%.




     Evans and Dennis (1973) described a test for mono-, di-, and tri-




ethylene glycols based on the reaction with sulfuric acid and potassium




permanganate.  This step oxidizes the glycols to aldehydes.  The latter




are converted with 3-methylbenzothiazol-2-one hydrazone hydrochloride




CMBTH) to green cationic chtotnagens which are measured spectrophotome-




trically at 630 nm.  This method was used to determine low levels of




ethylene glycols in surface waters.  Recovery was in the range of 1-5




mg/£  with a precision of 7%.







      Chromatographic Methods





      In addition to chemical tests, chromatographic procedures have been




developed for determining glycols in various media.




      Reid and Ivery (1975)  developed a rapid procedure using pyrolysis-




gas chromatography for measuring ethylene or propylene glycol in serum,




gastric, and urine samples.  The sample was introduced into  the chamber




of a  pyrolyzer and flash vaporized at 270°C.  The vapors were swept into




a GC  column maintained at 250°C.  Recovery of glycol ranged  from 96-104%




at 50 mg/dl levels and 98-104% at 100 mg/dl levels.




     Holman and Mundy (1976) used gas chromatography (GC)  to measure




ethylene glycol in mouse plasma.  Normal constituents of plasma did not




interfere with GC peaks of  ethylene or propylene glycols.




      GLC determination of propylene glycol using Chromosorbiol as a column




packing, resulted in 95-108% recovery in cosmetic samples  containing




20.0-40.0 mg propylene glycol (Champion, 1970).






                                      69

-------
     A potential problem of using gas chromatography for determining




ethylene glycol is ghosting, a desorption process in which a small  amount




of solute which had been injected in the GC column is removed from  the




column by subsequent injections of a solvent that does not contain  this




solute (Spitz, 1972) .   As long as an ethylene glycol standard used  is




the same concentration range as that in the sample,  ghosting will not




be a problem.




     Belue (19741 described a high-pressure liquid chromatographic  method




for the separation and identification of related polyhydric alcohols




such as ethylene glycol, glycoaldehyde, glycerol, and isoerythritol;




methyl ethyl ketone-water-acetone was used as a solvent.
                                     70

-------
                          III.  BIOLOGICAL EFFECTS





     The biological effects of ethylene glycol, propylene glycols,  and




butylene glycols are considered separately in :the sections which follow.









A.  Ethylene Glycol




     1.  Humans




          a.  Acute Toxicity





     During the  1920's and 1930's, ethylene glycol was not recognized as




a potentially toxic material.   In 1917 Bachem drank 45 ml of ethylene gly-




col without any  noticeable effects and in 1927 Page drank 15 ml, also




without  ill effect.  Hanzlik  et al.  concluded in 1931 that ethylene glycol




was "comparatively innocuous  as a solvent for medicinals."  The first




recognition in  the U.  S.  that ethylene glycol can be hazardous when in-




gested was a report by a  physician that two men  died from drinking Prestone




antifreeze  (.Anon., 1930). Since that time, numerous cases have been  re-




ported.   Haggarty (1959)  attributed  40-60 deaths per year to ethylene gly-




col intoxication. In  most cases, ingestion of ethylene  glycol, usually




as antifreeze,  is accidental.  For example, Pons  and  Custer  (1946). des-




cribed 16 fatal cases  in  soldiers who drank antifreeze.   Gaultier  et  al.




 (1976) reported on six patients who, lost in  the  desert,  drank  a mixture




of antifreeze  in water.   Friedman et al.  (1962)  described four  cases  of




 toxicity in young adults  who  mistook ethylene glycol  for ethanol.




      In humans,  the  lethal dose of ethylene  glycol  is about  100 grams or




about 1.4-1.6  g/kg.   This is  equivalent to 100 ml of  100% ethylene gly-




 col  or 105-110 ml of commercial antifreeze (Lavelie,  1977).   With suppor-




 tive  therapy,  patients ingesting up  to  970 ml have been successfully treated




 (Gaultier et  al., 1976).






                                      71

-------
     The clinical course of ethylene glycol poisoning can be divided


into three stages (Herman et al.,  1957; Friedman et al.,  1962;  Parry and


Wallach, 1974):


     i)  Central Nervous System Manifestations.


     Within 0.5-12 hours after ingestion of ethylene glycol, the patient


appears intoxicated.  Nausea, vomiting, mild hypertension, tachycardia,


and low grade fever may occur during the early stages.   Coma, convulsions,


and death may follow with large doses.


     Laboratory tests may reveal a  normal hematocrit and moderate leu-


kocytosis (10,000-40,000/mm3), with polymorphonuclear cells predominating.


Acidosis is a frequent finding (serum bicarbonate <10 meq/£; anion gap


>20) due to organic acids, such as oxalic acid,  produced  during metabolism.


Hypocalcemia is usually observed,  possibly due to chelation of  the calcium


ion by oxalate to form calcium oxalate crystals.


    The urine usually contains oxalate crystals  4-6 hours after ingestion


and shows a low specific gravity with excess protein.  If death occurs


during the first 24 hours, the major change observed is  the presence of


calcium oxalate crystals in the kidney, brain, leptomeninges, vessel


walls, and/or perivascular spaces.


    ii)  Cardiopulmonary Failure


    Twelve to 18 hours after ingestion of ethylene glycol, cardiopulmon-


ary failure, accompanied by tachypnea,  tachycardia, mild  hypertension

                      e
and cyanosis, may occur.  According to Parry and Wallach  (1974),, recent


cases have not reported this stage, probably because of  rapid recognition


and treatment of acidosis and hypocalcemia.   Death during this stage can


be attributed to pulmonary edema,  cardiac dilation, and bronchopneumonia.
                                     72

-------
     iii)  Renal Failure





     If the patient survives the first two stages, renal impairment,




ranging from proteinuria to anuria, may ensue.  Oxaluria usually occurs




soon after intoxication.  Oliguria may occur approximately 12 hours after




intoxication.








     iv)  Other Changes




     Central hydropic degeneration or fatty metamorphosis of the liver




with focal necrosis has been observed in cases of ethylene glycol poison-




ing (Friedman et al., 1962) .  Smith (1951) reported inflammation of the




diaphragm and Friedman et al. (1962) found inflammation of skeletal mus-




cle.  Petechial hemorrhage of the gastrointestinal tract may occur (Levy,




I960) , but major gastrointestinal bleeding has not been reported (Parry




and Wallach, 1974).








          b.  Occupational Exposure





     Although many deaths have resulted from acute ingestion of ethylene




glycol, few cases exist of chronic occupationally-related toxicity.  Ethy-




lene glycol is a compound of low volatility (Table 2), so inhalation ex-




posure is usually not significant.  However, in one case, workers were




exposed to an aerosol generated from heating ethylene glycol.  Another




case involved dermatitis in a worker having skin contact with ethylene




glycol.








               1)  Aerosol Inhalation





    Troisi (1950) described exposure of electrolytic condenser factory




workers to ethylene glycol at elevated temperatures.  The operation
                                     73

-------
involved the manual spreading of a heated mixture (105°C)  of 40% ethylene




glycol, 55% boric acid and 5% ammonia on a strip of paper using a paint-




brush.  Of 38 women exposed in this manner for several  years,  nine fre-




quently both lost consciousness and had nystagmus while five had only




nystagmus.  Lymphocytosis was observed in five workers.  Affected workers




had been exposed to this procedure for 1-5.5 years.  No abnormalities




were found in the urine of any worker.  The attacks disappeared when the




process was mechanized with no further exposure to the  women.   According




to the author, there was no evidence to implicate boric acid or ammonia




as causative agents; further elaboration was not given.




    The Russian investigators Dubeikovskaya et al. (1973)  reported that




in the manufacture of electrolytic condensers, workers  are exposed to an




average level of 44.8 mg/m3 ethylene glycol Grange 17-96.2 mg/m3).  No




signs of toxicity were noted in exposed workers.







               2)  Dermal Exposure




     A 17-year-old male developed eczematous dermatitis four months after




working in a factory where eyeglasses were made (Dawson, 1976) .  He bur^




nished glass lenses by bathing the lenses in a fluid made of one part




ethylene glycol to three parts water.  Patch tests with 3% ethylene glycol




in ethanol were strongly positive at 72 hours; no reaction occurred with




ethanol alone.
          c.
              Controlled Studies
     A group of 20-30 year old male volunteer prisoners (number not given)




was exposed to aerosolized ethylene glycol for periods up to 28 days




(Harris, 1969).  Psychomotor, psychological, and psychiatric tests were
                                     74

-------
performed.  In addition, blood chemistry and excretion of urea and crea-

tinine were monitored.  Volunteers exposed to 25 ppm (63.5 mg/m3) were

unaware of the presence of ethylene glycol; no changes in any tests were

detected.  Volunteers exposed to 50 ppm (127.0 mg/m3)"for short periods

of time" were able to detect the presence of ethylene glycol; they tasted

its sweetness and experienced an irritation in the pharyngeal area.  When

the level was raised to 75 ppm (190.5 mg/m3), without their knowledge,

several volunteers were awakened from their sleep, unable to tolerate

the exposure.



      2.  Nonhuman Vertebrates
          a.  Absorption, Distribution, and Excretion

               1)  Rat


     McChesney et al.  (.1971) studied the biological fate of ethylene gly-

col in male albino rats  which received intravenous injections of 139

mg/kg  ^C-labeled  ethylene  glycol.  The excretion and tissue distribution

1, 4,  and 24 hours following treatment are shown in Table 23.  The output

of 14C02 amounted to about 1.5% of the dose hourly for four hours; by 24

hours, 14.4% of  the dose had been eliminated as C02.  Urinary excretion

in 24 hours totalled about 46% of the dose.  There was a wide distribution

of lifC in various tissues, as shown in Table 23.  The level of   C in the

liver remained relatively constant while the level in the body as a whole
                      «
decreased; this  probably reflects the entry of ethylene glycol metabolites

into cellular metabolism.

    The urinary  excretion by rats following oral dosing of ^C-labeled

ethylene glycol  in 0-8, 8-24, and 0-24 hour urine samples is shown in
                                     75

-------
                            Table  23
Tissue Distribution and Excretion of ^C-labelled Ethylene Glycol
(139 mg/kg, i.v.) in Rats (McChesney et al., 1971)
Excretion or
Material
analyzed
Body weight (E) 	
Dose of i^C (y C) 	
Time before sacrifice (hr) .
expired air
feces
urine
blood
bladder wall
heart
lungs
spleen
liver
kidneys
brain
ileum
colon
carcass
Recovery (% of dofee) ....
but ion
1
220
2.5
1
1.3
0.2
—
5.8
0.1
0.4
0.9
0.3
8.7
0.6
0.7
5.1
4.0
81.0
109
(% of
2
198
5.0
1
1.7
0.2
—
4.5
0.1
0.4
0.9
0.3
6.4
1.2
0.7
5.7
5.8
58.3
86
tissue distri-
dose) in
3
290
3.5
4
5.8
0.4
37-7
4.1
1.1
0.1
0.4
0.6
9.2
1.0
0.4
4.5
2.9
37.3
106
rat no.
4
280
2.0
24
14.4
1.1
46.5
5.0
0.1
0.5
0.8
0.7
8.5
1.3
0.2
4.9
3.3
13.3
101
Blood volume assumed to be 5.4%  of body weight.
                                76

-------
Table 24.  In 24 hours, an average of 56% of the llfC dose was excreted;

of this, 57% was unmetabolized.
                                 Table 24
        Urinary Excretion of ll+C-Labeled Ethylene Glycol by Male
                       Rats (McChesney et al., 1971}
            Time after              Percent of dose*
             ,, ,     ,   ,         excreted as
            ethylene  glycol   	
            administration        .. ,                ,
                0-8              27.8 ± 1.0     45.9 ±0.3

                8-24             3.8 ± 0.5       9.8 ± 2.7

                0-24             31.7 ± 1.5     55.7 ± 2.4
                   dose of 1 ml/kg orally, as the labeled com-
            pound.

                   Data expressed as mean ± SEM; results from
            three  rats.
     The  approximate  ratio of glycolic and oxalic acids in  the 0-8 hour

urine was about 13:1  by weight; in a pooled  8-24 hour sample, the ratio

was about 8:30.  This is in  contrast to the  monkey, in which the ratio

is about  80:3 in 8-24 hour urine  (Section 2-a-4). ; the monkey converts

less ethylene glycol  to oxalate than does the  rat.

     Gessner et al. (1961) found  that in the albino rat, excretion of

ltf C-labeled ethylene  glycol  at 24 hours was  21% of a 0.1 g/kg dose and

58% of a  1.0 g/kg dose; ^C02 in  the expired air decreased  from  23%  of
                                     77

-------
the dose in 24 hours at 0.1 g/kg to 2.4% in 12 hours at 10 g/kg.




     In Sprague-Dawley male rats given an i.p. injection of 4 ml/kg (4.4




g/kg) of ethylene glycol, 1.50 ml/kg (1.66 g/kg) was recovered in the




urine during day one and 0.35 ml/kg (0.39 g/kg) was recovered on day two,




(Peterson et al., 1963).







               2)  Rabbit





     In Chinchilla rabbits given 0.1-2.0 g/kg of ^C-labeled ethylene




glycol, orally or by i.v. injection, about 20% of the radioactivity was




excreted-in the urine in two days (Gessner et al., 1971).  At doses of




2.5 and 5.0 g/kg, radioactivity in the urine increased to about 50%.




Gessner et al. (1971) identified the following metabolites in three rabbits




given 25 rag/kg orally:  oxalic acid (0.01-0.11% of dose), urea (0.65-1.5%




of dose) and unchanged ethylene glycol C6.0-15.1% of dose).








               3)  Do_g





     The blood glycol content in two dogs (breed and sex not reported)




given 2 ml/kg ethylene glycol (2.2 g/kg) by i.v. and oral routes is shown




below (Hanzlik et al., 1939a):
Blood glycol, mg%

2 hours
4 hours
6 hours
i.v.
337
270
202
gastric
310
283
240
               4)  Monkey





     McChesney et al.  (1971) studied the metabolism of ethylene  glycol








                                     78

-------
in the rhesus monkey (Macaca mulatta).  In one experiment, two females


received an intravenous injection of ^C-labeled ethylene glycol.  The


excretion and tissue distribution of label at one and four hours are

shown in Table 25.  After four hours, about 10% of the label had been


eliminated in the urine and 5% in expired air; only 0.1% had been found

in the feces.  At four hours, the highest proportion of ^C was found


in the carcass (69%), which includes muscle, skin, and bone, in the liver

(6.3%) and in the blood (4.7%).  The rest of the ethylene glycol was


widely distributed  (Table 25).


     In another experiment, the plasma half-life and rate of urinary


excretion were determined.  Unlabeled ethylene glycol was given to one

male and two females (1 ml/kg or 1.1 g/kg) by stomach tube.  A peak blood


level of 125 mg/100 ml was reached at 1-2 hours.  The plasma half-life


was 2.7-3.7 hours.

    The urinary excretion of ethylene glycol in three female rhesus

monkeys is shown  in Table 26.  In the period 0-24 hours, 23.5% of the


dose was excreted as unchanged ethylene glycol; no additional ethylene


glycol was excreted at 24-48 hours.  During the period 0-48 hours, other


metabolic products accounted for a mean of 22.2% of the dose.  Most of


the metabolic products was excreted in the 8-24 hour period.


     In a preliminary study of metabolism in chimpanzees, two females


were given ^C-labeled ethylene glycol intravenously (McChesney et al.,

                      c
1971).  One female given 2 ml/kg (2.2 g/kg), died after nine hours; 4%


of the 14C was identified in the urine at nine hours (40% of this was as


ethylene glycol).  The other female was given 1 ml/kg (1.1. g/kg).  During


the first eight hours, 28% of the administered label was excreted  (.30%
                                     79

-------
Excretion and Tissue Distribution of lkC Labeled Ethy-
         lene Glycola in Female Rhesus Monkeys
               (McChesney et al., 19711
Material
analyzed

Time before sacrifice (hr) .
expired air
feces
urine
b
blood
bile
bladder wall
heart
lungs
spleen
liver
kidneys
brain
adrenals
stomach
ileum
colon
carcass
Recovery (% of dose).
Excretion or tis-
sue distribution
(% of dose) in
monkey no .
1
3.3
41.3
1
1.3
0.1
4.3
5.4
0.02
0.1
0.9
1.6
0.2
4.5
0.8
2.8
0.02
0.9
3.7
2.7
78.8
108
2
3.3
41.3
4
5.0
0.1
10.4
4.7
Q.Q4
0.1
0.7
1.2
0.2
6.3
0.7
2.6
0.02
0.5
3.5
2.6
69.0
108
 a!39  mg/kg  administered  i.v.

 bBlood volume  assumed  to be 5.4%  of body weight.
                           80

-------
                                 Table 26
             Urinary Excretion of Ethylene Glycol and ltfC by
                     Female Rhesus Monkeys (McChesney
                               et al., 1971)
Time after
ethylene glycol
administration
(hr)
0-8
8-24
24-48
0-48
Percentage of
ere ted as
ethylene
glycol
17.9 ± 2.9
5.6 ± 2.3
0
23.5 ± 1.3
dose ex-
:,cb
25.9 ±5.1
19.0 ± 4.6
0.8 + 0.4
45.7 ± 4.9
                  SDose 1 ml/kg (1.1 g/kg) orally, as the
             labeled compound.

                   Data expressed as mean ± SEM; results
             from three monkeys.
as ethylene glycol).  In the 8-24 hour period, an additional 11% of the

1(tC was excreted (17% as ethylene glycol) .

     Squirrel monkeys (Saimiri sciurea, 450-750 g) were given an i.p.

injection of 3.2 ml/kg (3.5 g/kg) ethylene glycol (Peterson et al., 1963).

By 24 hours after the injection 0.20  (S.D. = 0.09) ml/kg  (Q.22 g/kg) of

the dose had been recovered in the urine.  All died 14-30  hours after
                      c
ethylene glycol administration.



          b.  Metabolism


     The pathway of ethylene glycol metabolism in the  rat  is shown in

Figure 3.  Ethylene glycol is oxidized to glycolaldehyde,  which is further
                                     81

-------
oxidized to glycolate.   Glycolate is metabolized to oxalate and C02 and

also to glycine and then serine (Chou and Richardson, 1978}.  Numerous

studies were conducted  to elucidate this pathway and to identify enzymes

catalyzing the reactions.  Some of the more recent major papers are
CH2OH CHO 	
CH2OH CH2OH
ETHYLENE GLYCOL GLYCOL-


COOH
CHNH,

CH2OH
SERINE
ALDEHYDE


G COOH F
	 i .
CH2NH2
GLYCINE
COOH
CH2OH
GUTCOLATE

C


/
/,
COOH *
— i 	
CHO
GLYOXYLATE
co2
/
/
D
i
4 COOH
— » i
COOH
0X4 LATE
            Figure 3.  Pathway of ethylene glycol metabolism
            in the rat (.Chou and Richardson, 1978).

                 The enzymes which catalyze the lettered re-
            actions are:  A. alcohol dehydrogenase; B. al-
            dehyde dehydrogenase; C. glycolic acid oxidase and
            lactic dehydrogenase; D. 2-oxoglutarate: glyoxy-
            late carboligase; E. xanthine oxidase, lactic de-
            hydrogenase, and glycolic acid oxidase; F. alanine:
            glyoxylate aminotransferase, and ornithine:glyoxy-
            late aminotransferase; G. serine hydroxymethyl-
            transferase.
 summarized in this section; in these papers, matabolites were isolated

 and possible toxic products were identified.
                      C

     Although some investigators have attributed the acute toxic effects

 of ethylene glycol to the alcohol itself (Pons and Custer, 1946; Rowe

 1963), the majority have attributed these effects to the metabolites.

 Milles (1946) suggested the metabolite oxalate to be the toxic
                                     82

-------
product, but other  investigators (Wiley et al., 1938; Roberts and




Seibold, 1969; McChesney et al., 1972) have suggested that oxalate plays




only a minor role.  Others have implicated the oxidation products of




ethylene glycol, such as glycolaldehyde, glycolate, and glyoxylate (Bach-




mann and Golberg, 1971; Richardson, 1973).  Recently, unequivocal evi-




dence has been presented that the metabolite glycolate is the specific




toxic agent in ethylene glycol poisoning in the rat (Chou and Richardson,




1978) and the monkey (Clay and Murphy, 1977).




     There appears to be a sex-related difference in the metabolism of




ethylene glycol.  Richardson  (1965) found that the oxidation of glycolic




acid to glyoxylic acid, and of glyoxylic acid to oxalic acid occurs twice




as rapidly in liver homogenates of male rats compared to female rats.  In




male rats, the level of glycolic acid is 30% higher than in females (Ri-




chardson, 1964).  Male rhesus monkeys are more sensitive to ethylene gly-




col than females (Roberts and Seibold, 1969) .








               1)  Rat





    Richardson (1973) showed that the toxicity of ethylene glycol is likely




due to metabolic products such as oxalate and glyoxalate.  Male Wistar




rats (140-160 g) were partially hepatectomized (about 1/3 or 2/3 of the




liver was removed) and maintained on a vitamin B,-dificient diet (to en-




hance urinary oxalate excretion).  An oral dose of 2 ml of ethylene gly-




col resulted in fewer deaths in hepatectomized than in intact rats.




Urine was collected for 48 hours and assayed for oxalate and glycolate.




Hepatectomized rats given ethylene glycol showed no change in the level




of urinary oxalate.  Liao and Richardson (.1972) showed that isolated per-




fused rat liver is capable of oxidizing ethylene glycol to oxalate.
                                     83

-------
Since removal of part of the liver,  which would decrease the metabolic

rate of ethylene glycol, lowered the toxicity of ethylene glycol,  the

authors infer that this glycol is not toxic in itself.  Rather,  oxidation

products such as oxalate and glyoxylate are likely responsible.

    Chou and Richardson (1978) demonstrated that the metabolite glycolate

is the specific toxic agent in acute ethylene glycol poisoning in Wistar

rats.  They administered alcohol dehydrogenase inhibitors, pyrazole and

4-methylpyrazole, prior to or with a lethal dose (10 ml/kg or 11.1 g/kg)

of ethylene glycol.  Alcohol dehydrogenase catalyzes the metabolic conver-

sion of ethylene glycol to glycolaldehyde.  The resulting mortality and

urinary metabolites were determined.  As shown in Table 27, administra-

tion of 3 mmol pyrazole/kg eight hours before or at the time of ethylene

glycol administration, reduced mortality from 100% to 0%.  Pyrazole was

less effective when given 4-12 hours after ethylene glycol.  The amount

of ethylene glycol and glycolate excreted in the urine varied with the

time of pyrazole administration; the excretion of glycolaldehyde, gly-

oxylate, and oxalate remained relatively constant (Table 27).  In addi-

tion,  the mortality varied inversely with the amount of ethylene glycol

in the urine and directly with the duration of pyrazole treatment, and the

amount of glycolate in urine.  Similar results were obtained when 4-methyl-

pyrazole was administered.

     Chou and Richardson (1978) also administered to rats other enzyme
                      c
inhibitors, in addition to pyrazole.  Inhibitors of the enzyme catalyzing

the  initial step in the metabolism of ethylene glycol, glycolaldehyde,

glycolic acid, and glyoxylic acid were given, as  follows:
                                     84

-------
                                                       Table 27
oo
Effect of Pyrazole on Ethylene Glycol Toxicity and Metabolism in the Rat"
(Chou and Richardson, 1978)
Pyrazole
(3 mmol/kg)
administra-
tion time
compared to
ethylene
glycol a
feeding
(hr)
c
-8
-4
0
+4
+6
+8
+10
+12
Mortality
(dead/
total)
6/6
Q/2
0/6
0/6
1/6
2/6
2/6
3/6
4/6
Content in 48-hr urine collection
Ethylene
glycol
(% recovery)
d
66 ± 4
86 ± 10
88 ± 9
73 ± 22
66 ± 16
44 ± 8
63 ± 1
47 ± 11
Gly col-
aldehyde
(mg/100 g)
—
2.0 ± Q.8
4.0 ± 0.9
3.2 ± 0.4
3/4 ± 0.9
4.2 ± 0.7
4.2 ± 1.2
3.5 ± 1.1
3.8 ± 0.5
Glycolic
acid
(mg/100 g)
—
37.9 ± 15.9
17.4 ± 2.6
20.2 ± 6.1
73.3 ± 12.6
96.7 ± 18.8
72.2 ± 8.3
133.8 ± 20.7
143.7 ± 14.3
Glyoxylic
acid
(mg/100 g)
—
0.08 ± 0.03
0.10 ± O.Q2
0.15 ± 0.01
0.11 ± 0.02
0.17 ± 0.03
0.23 ± 0.08
0.21 ± 0.17
0.13 ± 0.07
Oxalic
acid
(mg/100
—
15.6 ± 0
9.7 ± 2
9.3 ± 0
12.7 ± 1
17.6 ± 1
11.1 ± 2
16.3 + 1
7.0 ± 5
g)

.4
.3
.9
.7
.1
.1
.4
.1
             rat was fed 10 ml of ethylene glycol/kg.
        Values are given as the mean ± SD.
        No pyrazole administered.
        Control rats all died within 48 hours.

-------
       Substrate
       (g fed/kg)
Inhibitor
(administered/kg)
Mortality
within 48 hr
(dead/total)
       ethylene glycol    pyrazole (mg)

            8.5                0.0

            8.5              204.0
        glycolaldehyde

            3.0

            3.0


        glycolic acid

            5.0

            5.0


        glyoxylic acid

            2.0

            2.0
butyraldoxime (mg)

    0.0

  130.0

DL-phenyllactate (g)

    0.0

    3.0

DL-phenyllactate (g)

    0.0

    3.0
    3/6

    0/6




    3/6

    6/6




    3/6

    6/6




    3/6

    6/6
Only pyrazole, an inhibitor of alcohol dehydrogenase which catalyzes

the initial oxidation of ethylene glycol,  prevented deaths from ethylene

glycol.  Thus, the lethal toxicity is not  due to  ethylene glycol per se.

The lethal toxicity of the metabolites, however,  was dependent on the

compounds themselves; enzyme inhibitors lowered the LDsg  values.

     Chou and Richardson (1978) also determined the level of metabolic

intermediates in the plasma of rats following a dose of [U-^C] ethylene

glycol.  The major constituents were ethylene glycol and glycolate;

small amounts of oxalic acid were detected.  The level of ethylene glycol
                                     86

-------
By 48 hours, 34.2% of the label was excreted in the urine; in. another




experiment, three monkeys averaged about 46% llfC.  After unchanged




ethylene glycol, the next most important excretory product was glycolic




acid; little oxalic acid was present.  The authors consider that the




unidentified compounds included  primarily glycolic acid and also hip-




pur ate .




     In another study, McChesney et al. (1972) investigated the meta-




bolism of  glycolic and glyoxylic  acids in the rhesus monkey to further




elucidate the metabolism of ethylene glycol.  Two females weighing 3 kg




were given an oral dose of 500 mg/kg l-lkC glycolic acid; two other fe-




males weighing 3.5 kg were given 500 mg/kg [l-lt+C] glyoxylic acid.




Urinary excretion of label accounted for 37-52% and 34-69% of the admin-




istered glycolate and glyoxylate, respectively, within 96 hours.  Fecal




excretion averaged about 1% following administration of glyoxylate and 3%




following glycolate.  Individual labeled acids in the urine were then




analyzed.  Following a dose of 500 mg/kg glycolate, 34-44% of the dose




was excreted unchanged, 0.3-2»2% was excreted as glyoxylate, 0.3% as




hippurate and 0.3-1.3% as oxalate; 6% was not accounted for by these




acids.




     With a dose of 500 mg/kg glyoxylate, 24-59% was excreted unchanged,




0.1% as hippurate, 3% as oxalate and 2% as glycolate; 7-16% of the




urinary label was unaccounted for.  Urinary samples were similarly




analyzed in another experiment in which two females were given 60 mg/kg
                                    87

-------
decreased with time; the oxalate levels increased with time..  Glycolic




acid concentration remained constant.  These data appear below:
Sacrifice
time
(hr)
2
4
6
8
10
Percentage of dose recovered in plasma
Ethylene Glycolic Oxalic
glycol acid acid
2.48 0.71 6.70 x lo"3
1.81 0.61 5.13 x 1Q-3
0.70 0.60 3.55 x KT2
0.34 0.79 3.12 x 10"1
0.22 0.73 4.76 x IQ"1
                .2)   Monkey




      The rate of urinary excretion of metabolites of ethylene glycol




 in a female rhesus monkey,  who received an oral dose of 1 ml/kg




 (1.1 g/kg)   C-labeled ethylene glycol is shown below (McChesney




 et al., 1971):
Tine of
ethylene glycol
administration
(hr)
0-8
8-24
24-48
0-48

glycolic
acid
7.8 ;
3.1
0.5
11.5
Percentage
oxalic
acid
0.09
0.15
0.03
0.27
of dose
ethylene
glycol
15.1
2.0
tr
17.1
excreted
^C
25.4
7.9
0.9
34.2
as
unidentified
compounds
2.4
2.7
0.4
5.5
          of 1 ml/kg (1.1 g/kg) orally as the l^C-labeled  compound.
                                     88

-------
glyoxylate.  Now, only 20% of the lkC appeared in the urine within 96




hours and only 1-1.5% of the dose was excreted unchanged.  Oxalate was




the primary metabolite, accounting for 70% of the total label; hippurate




was a minor metabolite.  McChesney et al. (.1972) suggest that small




doses (i.e., 60 mg/kg) of glyoxylate can be metabolized efficiently,




but with large doses, such as 500 mg/kg, there is considerable loss by




excretion in the urine.




     Bachmann and Golberg (1971) hypothesized an adaptive shift in the




metabolism of ethylene glycol after prolonged exposure, based on prelimi-




nary results in rhesus monkeys exposed almost continuously to an




aerosol (600 mg/ta3) for up to 215 days.  Liver mitochondrial function




(measured as respiratory activity and oxidative phosphorylation) was




impaired after five months' exposure, but partial restoration of mito-




chondrial function was observed by months 6-7.  The number of monkeys




used was not specifically stated but apparently was small.  The authors




suggest two hypothetical adaptive processes:   .a) one involving easier




disposal of glyoxylate by reactions avoiding condensation with keto-acid




intermediary metabolites or  b) one in which another pathway ethylene




glycol metabolism assumes greater importance, by-passing glyoxylate.







          c.  Acute To xi city




               1)  Oral Administration




                    a)"  Lethal Dose Values
     The acute oral lethal dose (LDsg) averaged 8.3-15.3 g/kg in mice,




6.1-8.54 g/kg in rats and 6.61-8.1 g/kg in guinea pigs  (Table 28).
                                     89

-------
                                                     Table 28
VO
O
Oral Lethal Dose Values for Ethylene Glycol.
Species/ strain
mouse/white

mouse/NR
mouse/NR

rat/NR M&F

rat/Wistar

guinea pig/NR

guinea pig/NR

Sex/ No.
NR/4 per dose

NR
M&F/10-20
per dose
M&F/ 7-20
per dose
M/10 per dose

M&F/5-10
per dose
M&F/10 per
dose

Reported
value
7.5 cm3/kg

13.79 cm3/kga
13.1 ml/kg

5.5 ml/kg

8.54 g/kgb

7.35 ml /kg

6.61 g/kgb

LD50


_. . _. Converted value
Deviation , ,, N Reference
(g/kg)
8.3

15.3
range: 14.5
11.8-14.6
range: 6.1
5.00-6.05
95% C.L.; 8.54
7.31-9.99
range: 8.1
6.53-8.27
95% C.C.: 6.61
5.06-8.63
Latven and Molitor,
1939
Fisher et al.
Laug et al . ,

Laug et al. ,

Smyth et al . ,

Laug et al. ,

Smyth et al. ,


, 1949
1939

1939

1941

1939

1941

      NR = not reported.

      a
            determined after 24 hours .


            determined after 14 days.

-------
                    b)  Signs





     Laug et ai. (1939) administered large oral doses  of  ethylene  gly-




col to mice, guinea pigs and rats; the sex and strains used were not




reported.  Signs of intoxication included weakness,  loss  of muscular




coordination, prostration, coma, and death.




     The clinical signs of dogs dying from ethylene  glycol ingestion




are summarized in Table 29.  During the initial stages of intoxication,




signs of ataxia, tachycardia, hyperpnea, and tachypnea were consistently




observed.  Terminal signs included bradycardia and coma (Kersting  and




Nielson, 1966).  No sexual differences were apparent;  younger dogs of




either sex were less affected than older dogs.








                    c)  Tissue and Organ Changes.





     Mice, rats, and guinea pigs of unreported strain  or  sex  given




fatal oral doses of ethylene glycol showed pulmonary congestion and




hemorrhage (Laug et al., 1939).  At larger doses,  the  stomach was  also




hemorrhagic.  Microscopic changes included hydropic  degeneration of




cells lining the cortical convoluted tubules in the  kidney and focal




necrosis of the liver.




     Pulmonary edema and hyperemia and also gastric  or intestinal




mucosa hyperemia were observed in most dogs (mixed breeds) receiving




acute fatal oral doses of ethylene glycol (Kersting  and Nielson, 1966).




Calcium oxalate crystals were found throughout the renal  tubules and




the renal cortex.  Occlusion of the tubules was usually observed.   In




five dogs recovering from a nonfatal dose of ethylene  glycol  (4.4-6.6




ml/kg or 4.9-7.3 g/kg), biopsies revealed the presence of oxalate
                                    91

-------
                                Table 29
Signs of Intoxication in Ten Dogs (Mixed Breed) Dying
from Ingest ion of Ethylene Glycol (Kersting
and Nielson, 1966)a
Signs
ataxia
hyperpnea
tachycardia
tachypnea
anorexia
polydipsia
depression
emesis
miosis
bradycardia
coma
convulsions
death
Onset (hr)
2-4
2-4
2-4
2-4
6-35
6-35
10-40
intermittent
12-45
18-55
18-55
26-48
26-64
Frequency (%)
100
100
100
100
100
100
100
1
100
100
100
4
100
               Ten dogs of mixed breeds given 6.6-13.2 ml
          EG/kg (7.3-14.6 g/kg).
crystals in renal tubules at days 3-4; these crystals disappeared after
                      c
16-85 days.  In two dogs, no crystals were detected.  In all dogs, there

was no evidence of permanent damage or impaired renal function.
                    d)  Metabolic Acidosis
        Clay and Murphy (1977)  studied the effect of ethylene glycol
                                     92

-------
on metabolic acidosis in one male dog (17.7 kg) of an unreported strain.


A dose of 6 g/kg ethylene glycol was given via a nasogastric tube.


Severe metabolic acidosis occurred; from 0 to 5 hours after ethylene


glycol administration, the blood pH had decreased 2.4%,  blood P_.   had
                                                               CU2

decreased 27%, and blood HC02 had decreased 53%.  This is comparable


to acidosis observed in pigtail monkeys given ethylene glycol by injec-


tion (section 2-c-2) as in monkeys, acidosis in the dog  was caused  by


the accumulation of blood glycolate.  At five hours, 10.1 meq/1 glyco-


late were detected in the blood; prior to ethylene glycol administra-


tion, no glycolate was found.





               2)  Dermal and Ocular Application



     According to Rowe (1963) ethylene glycol produces no significant


irritation of the skin.  Severe prolonged exposure may result in a


slight macerating action.


     McDonald et al . (1972) evaluated the ocular toxicity of ethylene


glycol in New Zealand rabbits (1.5-2.2 g) .  Multiple topical or multi-


ple intraocular (anterior chamber) applications of a 0.40% or 0.04%


solution was nonirritating and nontoxic.  Solutions containing 4 and


40% applied to the eyes produced irritation, which consisted of chemosis,


swelling, and conjunctival redness.  All eyes were normal after seven


days.  No signs of systemic toxicity were observed.  Latven and Molitor


(1939)  reported that instillation of 0.5 ml of ethylene glycol into


the eye caused mild edema and hyperemia after 24 hours .





               3)  Parenteral Administration


                    .a)  Lethal Dose Values
     As shown in Table 30, the acute LDsQ values for ethylene glycol


                                     93

-------
                                         Table  30
Median Lethal Doses for Ethylene Glycol Administered Subcutaneously ,
Intravenously, or Intraperitoneally .
Species/strain
mouse/ Carworth
Farms
mouse/White
mo use /White
rat/Fischer
rat/Sprague-
Dawley

Sex/No . Route
-> F/60 i.p.
NR/4 i.v.
per dose
NR/4 s.c.
per dose
M&F/20 s.c.
per dose
M/10 per i.p .
dose
LD50
Reported
value
5.04 ml /kg
4.0 cm3/kg
6.0 cm3 /kg
5,300 mg/kg
5.8 ml/kga

_ . . Converted Reference
Deviation , , ,. ,
value Cg/kg)
S.E. = 1.52 5.6 Karel et al., 1947
4.4 Latven and Molitor, 1939
6.7 Latven and Molitor, 1939
95% C.L. = 5.3 Mason et al., 1971
3,857-7,478
6.4 Peterson et al . , 1963
NR = not reported.




 LDso determined after five days.

-------
in mice averaged 5.6 g/kg i .p., 4.4 g/kg i.v., and 6.7 g/kg s.c.  In




rats, the LD50 averaged 5.3 g/kg s.c. and 6.4 g/kg i.p.  These values




are comparable to those obtained after oral administration (Table 28).








                    b)  Signs





     Intramuscular or intravenous injection of ethylene glycol at doses




higher than 1.1-1.5 g/kg caused respiratory acceleration, depression,




and sometimes death to rabbits and white mice (Hanzlik et al.,  1931).




     Injection of 3-4 g/kg ethylene glycol in female pigtail monkeys




(Macaca nemestrina) resulted in transient narcosis, after which they




appeared normal but then worsened and became comatose ('Clay and Murphy,




1977).




     Intravenous injection of 1 ml/kg (1.1 g/kg) to one rhesus monkey




resulted in no behavioral changes or other signs; in another rhesus




2 ml/kg caused ataxia, then vomiting, with complete recovery after two




hours.  However, in 4 of 5 chimpanzees given 1, 2, or 6 ml/kg, (1.1,




2.2, or 6.6 g/kg), death occurred after 9.75-37.5 hours.  One chim-




panzee given 1 ml/kg, became ataxia and vomited, but recovered.




Oxalate crystals were found in the kidneys of all treated rhesus mon-




keys and chimpanzees at autopsy (Felts, 1969) .








                    c)  Urine Formation





     Wills et al. (196'9; meeting abstract) studied the effect of




ethylene glycol on the formation of urine by the cat.  In one experi-




ment, 1.5 ml/kg (1.7 g/kg) i.v. were administered during the study.




Ethylene glycol resulted in an increased production of urine, variable




changes in the clearance of creatinine and transtubular transport of
                                     95

-------
glucose, and decreased clearance of p-aminohippuric acid (PAH).   In a

second study, 0.5 ml/kg (0.6 g/kg)  i.p. ethylene glycol was administered.

In this experiment urine production was normal and the clearances of

creatine and PAH were reduced; transtubular transport of glucose and

sodium, but not PAH, were decreased.  The results suggest that ethylene

glycol interferes with blood flow through the nephron, possibly  by

partial blockage of the peritubular network of capillaries.



                    d)  Metabolic Acidosis


     Metabolic acidosis occurred in female pigtail monkeys (Macaca

nemestrina) given an intraperitoneal injection of 3 or 4 g/kg ethylene

glycol.  This effect was also noted for oral doses to dogs (section

2-c-l)-d) and was caused by the accumulation of blood glycolate  (Clay

and Murphy, 1977).



                    e)  Hematology


     Albino rats (150-200 g)  were given s.c. injections of 4 ml/kg

(4 .4 g/kg) ethylene glycol every other day for 30-65 days (Paterni et

al., 1956).  Progressive hemolytic and myelotoxic anemia were noted.

The reticulocyte count and hemoglobin index were normal but there was

a tendency to macrocytosis and granulocytopenia during the later stages

of intoxication.
                      t
     MacCannell  (1969) studied the effect of ethylene glycol and

1,2-propanediol on hemolysis or hemodynamic changes in 43 dogs (species

not given; 13.2-21.5 kg weight).  Either glycol was infused intra-

venously at 2% in saline (rate of 20 mg/kg/minute) for 2-5 minutes or
                                     96

-------
at 50% in saline (150-250 mg/kg/min) for 2-5 minutes.  At either con-




centration, both glycols caused an increase in superior mesenteric




blood flow and in cardiac output without any change in the cardiac con-




tractile force.  At the higher level, renal blood flow was  decreased




to a greater extent than it was at the lower level;  this decrease oc-




curred before the superior mesenteric flow increased.  Direct injection




of 25-50% glycol into the renal artery produced a decreased blood flow




through this vessel while direct injection into the superior mesen-




teric artery resulted in increased flow.




     These hemodynamic changes are partly a result of hemolysis which




occurred at levels of 20-50% glycols.  However, hemolysis did not




occur at the lower level tested (2%) but changes in blood flow were




still detected in the superior mesenteric blood flow.








          d.  Subacute Toxicity





     Subacute administration of ethylene glycol, orally or by injection,




has been used as a model to study renal hyperoxaluria or to induce




oxalate lithiasis (Fonck-Cussac et al., 1971; Kirschbaum and Bosmann,




1974; Lyon et al., 1966; Vaille et al., 1963; Tanret et al., 1962;




Gershoff and Andrus, 1962; Hammarsten, 1956).   A recent representa-




tive study (Kirschbaum and Bosmann, 1974) is discussed in the next




section.  Other subacute studies are available in which ethylene




glycol was administered by inhalation.








               1)  Oral Administration




                    a)  Renal Changes





     Kirschbaum and Bosmann (1974) induced hyperoxaluria in male
                                      97

-------
Sprague-Dawley rats (200-250 g)  by adding 1% ethylene glycol to the


drinking  water for up to 45 days.  After 15 days, kidneys from experi-


mental rats were significantly heavier than controls (1.42 g vs 1.30


g, dry weight) and had a significantly lower protein content (.69.2


mg/g vs 77.5 mg/g, wet weight).   However, by day 45, kidney weights


and protein content were not significantly different from controls.


Oxalate was identified in the urine of treated rats, indicating hy-


peroxaluria.  Ethylene glycol administration resulted in increased


urine activity of B-mannosidase, g-galactosidase, acid phosphatase,


N-acetylglucosaminidase, and N-acetylgalactosaminidase at weeks 3 and


6; these effects were more pronounced at week 3.  Also, cytidine mono-


phosphate-sialic acid synthetase activity was reduced on day 15 (but


not day 30).  Urine neuraminidase activity was elevated on day 15


but depressed on day 45 .


     Crystallization of calcium oxalate in the proximal tubules was


observed in 5 of 7 macaque monkeys (Macaca mulatta,  11.  irus, 11. radiata)


receiving a total dose of 15-132 ml/kg (.16.6-146.4 g/kg) over 6 to


13 days in  the drinking water (Roberts and Seibold,  1969) .  Of these


five monkeys, four were male and one female; the two not showing


crystallization were female, which is suggestive of a sex-related


difference.  In these five monkeys, the tubular epithelium adjacent


to the crystals was necrotic.  In the glomerulus, focal adhesions

                      c
and protein precipitate were noted.  Some distal tubules contained


protein precipitate, desquamated epithelial cells and leukocytes.


Epithelial  giant cells formed around the crystals in some cases.


There was no evidence of renal calcification.  Blood urea nitrogen
                                     98

-------
determinations revealed azotemia in treated monkeys.




                    b)  Calcium and Phosphorus Metabolism



     Rajagopal et al. (1977) investigated the effects of ethylene gly-


col on calcium and phosphorus metabolism.  Male albino rats (.100-110


g) were given 2 ml/kg/day (2.2 g/kg/day) for six days by gastric in-


tubation; this daily dosage is approximately one-fifth the LDsg.  The


level of bone and serum calcium was decreased, while urinary calcium


excretion was increased in treated rats.  In addition, serum phos-


phorus levels were increased while urinary phosphorus excretion de-


creased.  Serum alkaline phosphatase activity was increased.  Urinary


hydroxyproline was elevated and urinary citric acid was decreased.


These results indicate that bone demineralization may be an attempt


to maintain serum calcium concentrations during ethylene glycol intoxi-


cation .



                     c)   Blood Clotting


                                                                      _o
        Allen et al.  (1962)  administered 0.5 ml of ethylene glycol (10


  to  10~  dilutions) by stomach tube to SWR/J mice daily for 4 weeks.


   Every week, blood from treated and untreated mice was assayed for


   blood clotting defects (prothrombin time and thromboplastin generation


   time).  Clotting abnormalities occurred in 84.3% of ethylene glycol


   treated mice but in only 12.5% of untreated mice.  This study was


   prompted by the appearance of hemorrhage and death in mice whose


   bedding (pine shavings)  was sterilized with ethylene oxide.  Ethylene


   glycol is a breakdown product of ethylene oxide.  Allen et al. (1962)


   found a 62.3% incidence of clotting defects in mice given oral doses


   of extracts from ethylene oxide-treated shavings.  Reyniers et al.(1964)
                                     99

-------
   reported  on tumors  in mice exposed  to  ethylene oxide-treated bedding;


   this  is discussed in section III-A-2-f.
               2)  Inhalation Exposure


     Slight narcosis developed in rats exposed to 500 mg/m3  ethylene


glycol aerosol for 28 hours over five days (Flury and Worth, 1954).


No adverse effects were noted in mice or rats exposed to 350-400 mg/m3


eight hours/day for 16 weeks (Wiley et al., 1936).


     There were no signs of toxicity in rats, guinea pigs, rabbits,


dogs, or monkeys exposed to 10 or 57 mg/m3 ethylene glycol for eight


hours/day, five days/week for 90 days (Table 31) (Coon et al., 1970).


Continuous exposure of these species to 12 mg/m3 for 90 days resulted


in moderate to severe eye irritation in guinea pigs and rats; there


was apparent blindness in two of 15 rats after eight days.  Five


animals died during continuous exposure (Table 31).  For all three ex-


posure situations, results of hematological and histopathological ex-


aminations were comparable in treated and experimental animals (Coon


et al., 1970).



     Two chimpanzees  (sex and age not given) were exposed continuously to


 an atmosphere saturated with ethylene glycol aerosol (256 mg/m3) for


 28 days (.Felts, 1969).  Exposure was in a performance evaluation chamber,

                        t
 in which the animals had been previously trained in two tasks:  auditory


 discrimination, involving locating a source of sound, and visual dis-


 crimination, involving adjusting the length of one line to equal another.
                                     100

-------
                                 Table 31
              Mortality in Animals Inhaling Ethylene Glycol
                       Aerosol (Coon et al.,  1970)
Exposure, mg/m;
(Type) :
Ethylene glycol
10 .
(Repeated)
57 b
(Repeated)
12 c
(Continuous)
Control
Sprague Dawley
and Long Evans
rats
0/15
0/15
1/15
    aExposed in modified Rochester-type inhalation chamber.

     30 repeated exposures, 8 hours/day, 5 days/week.
    c
     Continuous exposure.
4/123
Princeton-de-
rived guinea pig
New Zealand
rabbit
Beagle dog
Squirrel monkey
0/15
0/3
0/2
0/2
0/15
0/3
0/2
0/2
3/15
1/3
0/2
0/3
0/73
0/12
0/12
0/8
 For  the  first  test,  reaction  time was  slower during the  second and third

 weeks of  exposure,  compared to pre-exposure times.  During week  four,

 latency was  decreased, 1sut not to that of pre-exposure response  time.

 For  the visual  discrimination task, one  chimpanzee maintained pre-ex-

 posure performance  levels.  Performance  in  the  other  monkey was  impaired

 during weeks three  and  four of exposure.
                                     101

-------
     Compared to pre-exposure levels, no major changes were detected




in serum chemistry or hematology tests, except for an increase in hemo-




globin and mean cell volume of red cells after exposure.  Urine concen-




trating ability, measured as serum and urine osmolality, was impaired




in both animals following exposure.  This condition is often associated




with dysfunction of the renal distal tubules.   Other renal tests were




within normal units (.insulin clearance, PAR clearance; PAH tubular max-




imum excretion).  The kidneys of only one chimpanzee contained oxalate




crystals.




    Four chimpanzees of unreported age or sex were exposed to an atmos-




phere of 256 mg/m3 ethylene glycol vapor for 28 days in a chamber simu-




lating a spacecabin atmosphere and altitude (.27,000 ft; 68% oxygen,




32% nitrogen; 5 psi) (Felts, 1969).  These tests were designed to eval-




uate the effects of a possible leak of ethylene glycol from the heat




exchanger system in spacecraft.   Few effects related to ethylene glycol




were detected:  total white counts cell were depressed two weeks after




exposure was terminated; the uric acid level was slightly elevated in




one animal.  No effects on other blood parameters were detected.  There




was no evidence of corneal erosion.
                                    102

-------
          e.  Chronic Toxicity


               1)  Oral Administration


     Seven groups of five white rats (50 g) were given water containing


ethylene glycol at 0, 0.5, 1, 2, 3, 5, or 10% (Hanzlik et al.,  1931).


Most rats receiving 2-10% died within 14 days.  Rats whose water con-


tained the lower percentages were maintained on this regime for 120-130


days.  No renal changes were noted in rats drinking 0.5% ethylene gly-


col but 4 of 5 rats receiving 1% ethylene glycol had oxalate crystals


in the urine.


     Three week old albino rats (12 males, 8 females) received 1 or 2%


ethylene glycol in the feed for two years (Morris et al., 1942).  In


six males, large urinary calculi were grossly observed.  Most test ani-


mals examined developed marked kidney damage and slight liver damage.


Changes in other organs were within the control range.


     The addition of ethylene glycol to the diet of two male rhesus


monkeys at a level of 0.2% and of one female at 0.5% for three years


produced no toxic effects (Blood et al., 1962).  Every three months,


x-rays were taken to detect possible calcification.  At the end of three


years, animals were sacrificed and necropsied.  No abnormal calcium


deposits were detected.  In the kidney of one male, the glomeruli had


thickened Bowman's capsules and were sclerotic; many tubules contained


granular eosinophilic material.  No significant changes were detected
                      c

in the other two monkeys.  One male and the female were obese.


    In three macaque monkeys (Macaca mulatta and irus) given 33-137


ml/kg (36.6-151.9 g/kg) in the drinking water over 13 to 157 days, renal


changes were proportional to the dose (Roberts and Seibold, 1969).
                                   103

-------
There was marked to extreme deposition of calcium oxalate crystals in




the proximal tubules; contiguous epithelial cells were necrotic.  Oc-




casional focal granulomas were observed in areas where tubular damage




was extreme.  In two monkeys, calcium oxalate crystals were also pre-




sent in the walls of cerebral vessels and adjacent tissues.




    In three monkeys receiving 17-28 ml/kg (18.8-31.0 g/kg), no renal




changes could be attributed to calcium oxalate deposition; however, these




monkeys showed mild glomerular damage and azotemia, suggesting a toxic




effect of ethylene glycol apart from its conversion to oxalic acid.







               2}  Inhalation Exposure





     Twenty mice and ten rats of unreported strains were exposed to




ethylene glycol (350-400 mg/m3) for eight hours a day, five days a week




for 16 weeks (Wiley et al., 1936).  Three mice and one rat died during




the experimental period.  The organs of nine rats and 17 mice were ex-




amined histologically; no pathological changes could be attributed to




the ethylene glycol exposure.  Some mice had a parasitic infection of




the liver; isolated cases of myocardial degeneration, testicular de-




generation and dermal ulceration were considered as "coincidental findings."




Some rats had an intestinal infection;  two showed moderate regressive




changes in the testes.








               3)  Paizenteral Administration





     No excess mortality occurred in 200 Fischer rats of both sexes re-




ceiving twice weekly s.c. injections of 0.03-1 g/kg ethylene glycol for




one year.  Overall mortality was 2% in both treated and control rats




(Mason et al., 1971).  As discussed further in section "f" on special studies,
                                   104

-------
tumor incidence was comparable in experimental and control animals.




     No growth retardation occurred among 80 male and female Fischer




rats given s.c. injections of 1 g/kg twice weekly for one year.  At




12 months, the final body weights ranged from 98-105% of the vehicle




(saline) and no-treatment controls (Mason et al., 1971).







          f.  Special Studies




               1)  Reproduction





     Ethylene glycol exhibited an intermediate order of toxicity when




injected into the yolk sac of fertile white Leghorn eggs prior to in-




cubation (Mclaughlin et al., 1963).  Injection of Q.05 or 0.10 ml re-




sulted in 20% or 35% mortality, respectively.




     Clegg (1964) reported 17-20% mortality in five day old Light Sussex




embryos injected in the yolk sac with 0.1 or 0.05 ml ethylene glycol.




A ten second immersion of a five-day incubated egg resulted in 60%




mortality (50% in water controls), while similar immersion of a f ive-




-day unincubated egg resulted in  20% mortality (25% in water controls).




Based on results from this and other compounds, Clegg (1964) concluded




that such tests are not reliable  indicators of toxicity because of vari-




able results.  Also, a clear-cut dose relationship could not be estab-




lished; such a relationship was only suggested in the immersion test,




but even water caused an unusually high incidence of abnormalities.




     Walker (1967) reported no mortality in three day incubated eggs




injected with 0.1 or 0.05 ml ethylene glycol into the yolk.




     Gebhardt (1968) tested the effect of a number of glycols on the




White Leghorn chick embryo.  Injection of 0.05 ml of ethylene glycol
                                    105

-------
into the air chamber resulted in 6% mortality on day 0 of incubation




and 37% mortality on day 4.  When this amount was injected into the




yolk sac, mortality was 5.6% by day 4.  In neither experiment were mal-




formations noted in surviving embryos.








               2)  Carcinogenicity





     Ethylene glycol was not carcinogenic when administered subcutaneous-




ly in day-old mice (WAKF,  1969) .   Groups of 50 male and 50 female Texas-




-Yale mice were given a single dose of either 1 or 10 mg ethylene glycol;




the higher dose is approximately the LDso.   Groups of 200 male and 200




females received saline (control). .  Animals were observed daily and




weights were recorded every 1-2 weeks.  At 15 weeks of age, a respira^




tory condition resulted in a moderate level of mortality in the popula-




tion.  Animals were sacrificed at 15 months.  Mice which were sacrificed,




or which had died, were examined grossly and histologically.   There




were no changes attributable to ethylene glycol injection.  Neoplastic




and non-neoplastic pathology were comparable among treated and control




animals.    Tumor incidence is summarized in Table 32.




     The tumor incidence, in rats receiving s.c. injections of 30-1,000




mg/kg ethylene glycol twice weekly for one year was comparable to that




found in controls, as shown in Table 33 (Mason et al., 1971).




      Homburger  (1968; unpublished studies cited in Johnson,1978  ">




 investigated the tumocigenic  effect of ethylene glycol.  Male C57BL/6




 mice were injected s.c. in the  groin with 26 mg ethylene glycol  in




 tricaprylin.  The number  and  schedule of doses were not reported; negative




 controls received tricaprylin.  Five weeks after  the injection,  the
                                    106

-------
                                Table 32
Evaluation of Carcinogenicity in Texas-Yale Mice
(WARF, 1969) a

I
1)




2)


II
1)




2)


Ethylene glycol
1 mg 10 mg
M F M F
Animals Dying During 19 14 23 17
Test;
benign neoplasms
lung adenoma 0000
adenoma or 0000
fibroma
skin papilloma 0000
hemangioma 0001
malignant neoplasms
sarcoma 0001
carcinoma 0001
Animals Sacrificed 00 01 00 00
., _ ZZ £-L £.£. t.3
at 15 mo .
benign neoplasms
lung adenoma 0010
adenoma or 0000
fibroma
skin papilloma 0000
hemangioma 0000
malignant neoplasms
sarcoma 0614
carcinoma 0100
Saline
M F
94 50

0 1
0 0
0 0
0 0

0 2
0 1
33 43
1 9
1 1
1 0
0 1

1 8
0 0
     one-day mice given s.c.  injection of 1 or 10 mg ethylene glycol;
surviving animals sacrificed at 15 months.
                                   107

-------
                                 Table 33
Tumor Incidence in
Injections

% tumor bearing rats
male
female
tumor location
male
injection site
other
female
injection site
mammary
uterine
other
Fischer Rats Given Twice Weekly S.C.
of Ethylene Glycol for One Year
(Mason et al. , 1971)
Ethylene
glycol

5
12


2/100
4/100

0/100
5/100
11/100
6/100
Saline
(vehicle
control)

6
14


0/50
3/50

0/50
3/50
5/50
8/50
No treatment
(negative control)

10
12


1/50
6/50

0/50
1/50.
5/50
7/50
     Doses used:  30, 100, 300, or 1,000 mg/kg; rats initially four
weeks old and weighed 60 g.
 injection  sites were  excised,  minced  and  injected  into  a  mouse of  the same

 strain  and age; these mice were  sacrificed  eighteen weeks after the

 transfer and no tumors were  detected.   In another  study,  females CF1  and

 A/He mice  were given  single  or repeated  (I/month for  7  months)  i.v.


                                   108

-------
injections of 26 mg ethylene glycol; negative controls received Ringer's


solution.  Mice were sacrificed after 28 weeks and examined for lung


tumors.  Compared to controls, there was no significant increase in lung


tumor incidence in ethylene glycol-treated mice.


     Reyniers et al. (1964) reported on hemorrhage in males and tumors in


female germfree albino mice exposed accidently to bedding sterilized


with ethylene oxide.  Ethylene glycol was identified in the bedding and


was implicated as a possible causative agent; however, this was not a control-


led study and other breakdown products of ethylene oxide or ethylene oxide


itself were not assayed for in the bedding.  As such, it is not possible


to identify this tumor-causing agent(s).  As discussed in section


III-A-d-1), Allen et al.  (1962) found clotting abnormalities in mice fed


ethylene glycol.  Reyniers et al.  (1964) reported that all males exposed


to the ethylene oxide-treated bedding died of massive hemorrhages, failure


to clot blood and jaundice.  Ninety percent of surviving exposed females


developed tumors.


     Berenblum and Haran  (1955) evaluated the carcinogenic effect of


urethane in ethylene glycol by skin painting in female Swiss mice.


Urethane controls received a primary treatment with  ethylene glycol


 (1,12  or 86 applications); some also received a secondary treatment with


croton oil or liquid paraffin  (70  applications).  No papillomas developed


in mice treated with ethylene glycol alone or with liquid paraffin.  One


mouse  in each group receiving ethylene glycol with croton oil  (out of  18-

                     t?
19/group which survived)  developed a papilloma; none of the mice  receiving

croton oil alone developed papillomas.


     Deringer (1962) also evaluated the carcinogenicity of urethane in


ethylene glycol by skin painting.  Strain HR/De mice were painted with
                                   109

-------
ethylene glycol as a control, twice a week for lifetime.  Tumor incidence




in ethylene glycol treated mice was comparable to that in untreated




mice.








               3)  Mutagenicity





     Embree (unpublished thesis cited in Johnson, 1978) tested a "small




amount" of ethylene glycol on Salmonella typhimurium strains TA 1535,




TA 1537, and TA 1538  in a point mutation study without activation; no




revertants were found.
3.  Aquatic Organisms




     Price et al. (1974) determined that the 24 hour median tolerance




limit (11 ) of brine shrimp (Artemia salina) to ethylene glycol exceeds




20,000 mg/1.




     Portmann and Wilson (1971) determined the LC50 of ethylene glycol




to be >100 mg/1 for Brown shrimp (.Crangon crangon) .  An unspecified




number of between 8-25 shrimp were tested in sea water; the LCso was




determined after 48 or 96 hours.




     Based on the data of Portmann and Wilson, Hann and Jensen (1974)




assigned ethylene glycol an aquatic toxicity rating of 1, a rating




corresponding to "practically non-toxic" (TL  96 ranges from 100-1,000




mg/1).  According to Hann and Jensen, ethylene glycol could pose a BOD




problem at sub-toxic concentrations.




     The 96 hour LCso for rainbow trout (Salmo gairdneri)  was more




than 18,500 mg/1 of reagent grade ethylene glycol  (Jank et al., 1974).
                                    110

-------
4.  Plants





     Ethylene glycol inhibited elongation of oat (Avena sativa)  coleop-




tile segments over the range of 0.5-3% in aqueous solutions or in in-




doleacetic acid (Farr and Norris, 1971).




     Treatment of maize microsporocytes with, ethylene glycol produced




aberrant chromosome behavior such that meiotic centromere-spindle in-




teractions were disrupted (Maguire, 1974).




     D'azEto(1948) found that exposing Allium cepa to 1-2 molar ethylene




glycol for 12-24 hours resulted in a lengthening of metaphase and a




vacuolization of meristematic cells.









5.  In Vitro Studies





     Preparations of rabbit Intestine and rat uterus were exposed in




vitro to ethylene glycol (Hanzlik et al., 1931).  At dilutions of 1:250




and 1:100, there was a gradual depression of contraction which recovered




upon washing.




     In vitro, ethylene  glycol at molar equivalents of 5% glycerol,




increased total cell lipid content in mouse fibroblast and human liver




cultures  (Mackenzie et al., 1968).




     Bachmann and Golberg (1971) studied the effects of ethylene glycol




and its metabolites, primarily glyoxalic acid, on mitochondria isolated




from the liver, kidney,  and brain of Sprague-Dawley rats  (40-50 g) ,




beagle dogs  (9 kg) and 'rhesus monkeys (Macaca mulatta, 9 kg); the  com-




pounds were  tested at concentrations up to 10 mM in vitro.   Ethylene




glycol itself had few adverse effects on mitochondria; respiration,




oxidative phosphorylation and citric-acid cycle activities were not
                                    111

-------
affected at any concentration tested while substrate-level phosphoryla^-




tion was inhibited at 10 mM.  In contrast, 1 mM glyoxylic acid inhibited




mitochondrial respiration from liver, brain, and kidney of rats (using




succinate, a-ketoglutarate or B-hydroxybutyrate as substrate)  and also




inhibited oxidative phosphorylation from these tissues (pnly when a-




ketoglutarate was used as substrate).  In addition, 10 mM glyoxylic acid




strongly inhibited two enzyme systems of the citric-acid cycle Ciso-




citrate dehydrogenase and a-ketoglutarate dehydrogenase.  In dogs,




glyoxylic acid inhibited liver and brain mitochondria using a-ketoglu-




tarate as a substrate.  In monkeys, glyoxylic acid inhibited respiration




of brain mitochondria with all substrates and oxidative phosphorylation




with succinate and a-ketoglutarate as substrates; only a slight effect




was noted with liver mitochondria exposed to glyoxylic acid.  These




data give further evidence that the toxic effects of ethylene glycol




are attributable to the formation of glyoxylate.
                                    112

-------
B.   Propylene Glycols





     The biological effects of 1,2- and l,3'-propylene glycols are dis-




cussed in separate sections which follow.








     1.  1,2-Propanediol




          a.  Humans




               1)  Acute Toxicity




                    a)  Oral Ingestion





     1,2-Propanediol is classed as a generally recognized as safe (GRAS)




food additive in the Code of Federal Regulations (see section IV).  It




is used in food as an emulsifying and plasticizing agent, humectant,




surfactant, and solvent (Griffin and Lynch, 1972).  The amount of 1,2-




propanediol that is added to certain foods is listed in Table 34, and




ranges from about 15% in seasonings and flavors to less than 0.001%




in egg products and soups.




     The Federation of American Societies for Experimental Biology




(FASEB, 1973) has estimated the possible daily intake of 1,2-pro-




panediol to be as follows:
Daily
Age group

0-5 months
6-11 months
12-23 months
2-65+ years
ave rage
mg
11
10
183
349
mg/kg
21
13
17
6
Daily
maximum
mg
81
333
594
1,380
mg/kg
20
42
54
23
                                    113

-------
Table 34
Level of 1,2-Propanediol
(FASEB, 1973)
Food category -

seasonings and flavors
sugar, confections
processed vegetables, juice
sweet sauce, toppings,, syrups
frozen dairy desserts, mixes
soft candy
baked goods, baking mixes
hard candy
chewing gum
alcoholic beverages
gravies, sauces
processed fruit, juices, drinks
non-alcoholic beverages
gelatins, puddings, fillings
meat products
milk, milk products
fats and oils
cheese
reconstituted vegetable proteins
snack foods
Used in Food
Propylene

glycol
usual use maximum use
percent
14.900
5.073
—
0.310
0.138
0.089
Q.079
0.072
0.068
0.066
0.049
0.049
0.038
0.036
0.023
0.023
0.014
0.007
0.006
0.002
percent
15.199
5.086
0.500
0.421
0.213
0.144
0.244
0.135
0.300
0.588
0.098
0.082
0.124
0.068
0.053
0.038
0.032
0.062
0.006
0.092
   114

-------
                            Table 34 (Continued)
                                                Propylene glycol
             Food category                 	     "
                                           usual use     maximum use
      poultry products                      0.001          0.010

      other grain products                  0.001          0.001

      eggs, egg products                   <0.001         <0.001

      soups, soup mixes                    <0.001         <0.001

      dairy product analogs
     Few reports of adverse effects of 1,2-propanediol ingestion have

appeared in the literature.

     Martin and Finberg (1970) reported a case of suspected 1,2-propane-

diol intoxication in a 15-month old boy receiving large doses of vitamin

C suspended in propanediol.  The patient received 250 mg vitamin C three

times a day (.the dose of the glycol was not estimated) .  By the eighth

day of this treatment, the child had an irregular apical heart rate

(sinus arrhythmia) and by days 10-13 had three episodes of unresponsive-

ness and tachypnea.  Blood glucose concentration during these episodes

was 70, 41, and 42 mg/100 ml.  Symptomatology disappeared when vitamin

C treatment was withdrawn.  The authors also reported on personal commu-

nication with two physicians who each noted stupor in a patient receiv-

ing about 60 ml 1,2-propanediol in a vitamin D preparation.  Recovery

was complete in a few hours.


                                    115

-------
     Gate and McGlothlin (1976)  described a case of 1,2-propanediol




overdose associated with lactic  acidosis.  A patient of unreported age




and sex ingested an unknown quantity of the glycol.  The patient Ini-




tially was comatose with metabolic acidosis and the following measure-




ments were made:  blood pH, 7.24;   CC>2, 11 meq/1;  anion gap, 29;  blood




lactic acid level, 18 meq/1; blood level of 1,2-propanediol 70 mg/dl.




The patient responded to bicarbonate therapy.  It is known that 1,2-




propanediol is converted to lactic acid in the liver and then meta-




bolized to pyruvic acid.  The authors suggest that the lactic acid in




this patient was rapidly metabolized to pyruvic acid with disappearance




of the metabolic acidosis.







                    b)  Dermal Exposure





     Seidenfeld and Hanzlik (1932) reported that 1,2-propanediol re-




sulted in a burning sensation when applied to the tongue.  Other inves-




tigators have reported on the potential for allergic reaction after skin




application.




     Warshaw and Herrmann (1952) patch tested the skin of 866 volunteers




for allergic reactions to undiluted 1,2-propanediol.  These volunteers




were patients at a clinic treating dermatologic conditions.  Sixteen




percent (138) of the patients showed a positive occlusive patch test;




positive reactions ranged from simple erythema to erythema with indura-




tion and vesiculation.  However, when an "open" test (i.e., no patch)




was used in 16 patients showing  a positive reaction in the "closed" test,




no reaction was noted.  The authors suggest that excessive dehydration




of the skin might predispose subjects to the reactions.  They further







                                    116

-------
suggest that skin-sensitivity to propanediol has not been widely re-




ported because most uses of the glycol are without an occlusive dressing.




     Using a modified Draize test, Phillips et al. (.1972) applied un-




diluted USP 1,2-propanediol to an occluded area on the forearm of four




male volunteers.  Examination at 24 and 72 hours revealed no irrita-




tion.  Furthermore, no irritation was observed in a 21-day continuous




patch test using 1-80% propanediol (in water), and a 21-day open test-




ing system using 50 or 100% propanediol.




     1,2-Propanediol failed to produce contact sensitization or skin




irritation in the Draize test using 204 male volunteers  (Marzulli and




Maibach, 1973).  One half gram of a 12% preparation in a creme vehicle




was applied to  the arm with an occlusive patch and removed after 48-72




hours.  Two weeks later, the test material was applied again in the




same manner.  No adverse effects were recorded.




     Pevny and  Uhlich (1975) calculated a "sensitivity index" for 1,2-




propanediol based on the percentage of subjects showing a positive skin




test reaction.  In three studies, 40 of 778 patients tested (,5.1%).




showed a positive reaction.




     Shanahan (1977) evaluated the dermatopharmacologic activity of




1,2-propanediol in humans.  This compound was a mild skin irritant fol-




lowing patch test exposure.  A synergistically enhanced irritation po-




tential was observed with triethanolamine-stearate, a cosmetic emulsi-




fier.  This combination resulted in progressive desquamation of the




stratum corneum.  Detection of irritation was influenced by the use




of a semi-occlusive rather than a fully occlusive patch.  Shanahan (.1977).




suggests that the conflicting evidence for the allergy/irritation poten-




tial of propanediol alone and in cosmetic preparations is due to several





                                    117

-------
factors.  1) The degree of occlusivity of the patch test used;  2) the




concentration of propanediol used;  3) presence of surface active agents




in preparations tested; and  4) the influence of environmental tempera-




ture and humidity.







               2)  Controlled Studies





     In three subjects given oral doses of 1 ml/kg (.1 g/kg) 1,2-propane-




diol, the maximum concentration of blood glycol was measured after 0.5




hour (Hanzlik et al., 1939a).  This level persisted for about four hours,




but individual variation occurred (Table 35).  After ten hours, about




20-25% of the dose had been eliminated in the urine.  1,2-Propanediol




was also detected in the saliva.










          b.  Nonhuman Vertebrates




               1)  Absorption, Distribution and Excretion





     In a study on 48 cats, 72 rats, and 16 rabbits, Van Winkle and



Newman (1941) demonstrated that absorption of 1,2-propanediol was




particularly rapid from the jejunum.  Ten ml/kg (10.4 g/kg) were injected




directly into the jejunum,  duodenum, colon,  or stomach^  Data were presented




for cats and were as follows:
                                   118

-------
                                 Table 35
     Amount of 1,2-Propanediol in the Blood, Saliva,  and Urine of Three
              Subjects  Following an Oral Dose of 1 ml/kg.
                          (Hanzlik et al., 1939a)
           Blood, mg%            Saliva, mg%             Urine,  g
Hours  —	
0
0.5
1
2
3
4
6
7.5
8
10
0 0
174 79
158 95
95 111
79 111
111 79
47

79 16

0 0
47.4
31.6
47.4
63.2
63.2
15.8
95
15.8

00 00 0
142.2
126.4
142.2 1.86
142.2
94.8 7.56 5.16
94.8
63 6.20 6.60
47.4 13.10
1.64
    a
     Sex not reported.
                                    119

-------

stomach
colon
duodenum
j ej unum

30
20
40
60
90
% Absorbed After
min 60 min
20
50
65
90

120 min
30
75
95
98
     Following i.v. injection of 1.0 g/kg 1,2-propanediol, the blood


level of the glycol in rabbits (strain and sex unreported). was 80 mg%


at 0.5 hour, 40 mg% at 2 hour, 16 mg% at 5 hour,  and 8 mg% at 1 hour


(Strack et al., 1960).  The elimination of propanediol was calculated


to be 0.366 mg/cm3 per hour.


     Lehman and Newman (1937) measured the level of 1,2-propanediol in


the blood for several hours after oral and intravenous administration


to intact and nephrectomized dogs.  As shown in Table 36, urinary ex-


cretion is quite important in the elimination of 1,2-propanediol; a


large fraction of the dose remained in the blood of nephrectomized ani-


mals.  In another experiment, Lehman and Newman (1937) estimated that


up to 45% of the total dose was excreted within 24 hours.
                      
-------
                                 Table 36
             Rate of Disappearance of 1,2-Propanediol in Dogs
                         (Lehman and Newman,  1937)
Hours after
dose
2
4
6
8
10
12
14
mg Propylene
Oral (12 ml)
Intact Nephrectomized
dog dog
14.5 17
12
11.5 16.5
10 15.5
8 15
glycol/ml blood
Intravenous (4 ml)
Intact Nephrectomized
dog dog
5 6
3 5.5
5
4
2
0.5 hours with a 2 ml/kg dose.  However,  with the 8 and 12 ml/kg doses,

4-8 hours were required for complete absorption.   The authors speculate

that absorption through the stomach is slight, as 124 mg% 1,2-propanediol

introduced into an isolated stomach was only slightly absorbed.  Also,

absorption of the glycol was delayed in anesthetized dogs which, accord-

ing to the authors, was likely a result of depressed gastric motility.


                                    121

-------
     Weil et al.  (1971)  administered a single dose of 5  g/kg  1,2-propane-




diol to female beagle dogs previously treated with 2  or  5  g/kg daily




for up to two years.   Peak levels (0.56%)  of plasma propanediol were




reached after 6-8 hours.  Levels  were halved by  12 hours.   In dogs  re-




ceiving daily doses of 5 or 2 g 1,2-propanediol,  a maximum blood value




of 0.2% (V/V) was recorded, but all other  individual  values were below




0.1% (V/V).







               2)  Metabolism





     Ruddick (1972) reviewed the  biochemistry of  1,2-propanediol and




suggested that the oxidation of this compound to  lactic  acid  or pyruvic




acid follows one  of two  pathways  depending on whether the  substrate is




the free glycol or the phosphorylated glycol (data primarily  from Rudney,




1954 and Miller and Bazzano, 1965).  Free  1,2-propanediol  is  metabolized




through lactaldehyde, methylglyoxal, and lactic  acid  as  shown below:
           -2H+          -2H+
3I
OH
1,2-pro-
panediol
	 CH3CHCHO —
1
OH
lactalde-
hyde
^ CH3CCHO -^
||
II
0
me thy 1-
glyoxal
— *- CH3CHCOOH
1
OH
lactic acid
Phosphorylated glycol (1,2-propanediol phosphate)  takes  the  following




route:  acetol phosphate, lactaldehyde phosphate,  lactyl phosphate,




and lactic acid, as follows:
                                       122

-------
               -H"
CH3CHCH2OP03H2

   OH
1,2-propanediol
phosphate
                                   lTl2(-\                _ Oil
                                       >  CH3CHCHOP03H2 -==»
                     acetol
                     phosphate
                                            OH.
                                              OH
lactaldehyde phosphate
CH3CHCOP03H2

   OH

lactyl
phosphate
                   CH3CHCOOH

                      OH

                   lactic acid
According to Rudney  C1954) , 1,2-propanediol phosphate occurs in rat

liver at 1-2% of the  total acid-soluble phosphorus in liver.

     Once 1,2-propanediol is oxidized to lactate or pyruvate, it can

provide energy by  further oxidation through the tricarboxylic acid cycle

or through the glycolytic pathway, the latter contributing to glyco-

gen  formation (Ruddick, 1972).  Hanzlik et al. (1939a and b) were the

first to report on the glycogenic action of 1,2-propanediol.  In starved

rats given oral doses of 0.5-15.0 ml/kg (0.5-15.5 g/kg), the amount of

liver glycol was increased compared to controls, as shown on the next page.

Total body glycogen was also increased following 1,2-propanediol in-

gest ion.              «

     Giri et al. (.1970) reported a significant increase in blood glu-

cose 15 minutes after i.p. injection of a near fatal dose of 1,2-pro-

panediol CIO ml/kg or 10.4 g/kg) to male Sprague Dawley rats.  Blood
                                    123

-------
                    Liver glycogen, mg%                     .
 Dose,	Maximum increase,

           controls     1 hr    2 hr    3 hr    4 hr      "8/8 liver
 0.5         148         211      156      208      135          0.63




 1.0         123         333      336      330      294          2.13




 2.0         113         241      371      465      720          6.07




 5.0         241         331      425      570      852          6.11




10.0          75          72      224      673      726          6.51




15.0         212         359      581      699      627          4.87
     aRats weighed 80-150 g;  strain  and sex not  reported;  five  rats/group;

      data from Hanzlik  et al.  (1939a  & b) .
 glucose reached a maximum by 90 minutes,  but remained significantly



 elevated (p < 0.01) by hour 6.  During this period,  liver glycogen re-



 mained unchanged, suggesting that the increase in blood glucose may



 have been due to conversion of the glycol to blood glucose, rather than



 to the mobilization of liver glycogen.  Twelve hours after propylene



 glycol administration, there was a severe depletion of liver glycogen.
                       c


      Whittman and Bawin (1974) presented data showing that glucose



 formation from 1,2-propanediol proceeds via phosphoenolpyruvate carboxy-



 kinase.  Fasted female Charles River CD rats (110-198 g) were injected



 i.m. with 200-2,000 mg/kg 1,2-propanediol during pentobarbital-induced
                                     124

-------
amnesia.  After 90 minutes, the concentrations of blood glucose and




liver glycogen were determined.  Propanediol caused a. dose dependent




increase in liver glycogen, blood glucose, and the rate of glycogen




synthesis.  In a time course study using 500 mg/kg propanediol injected




i.m. , the rate of synthesis of liver glycogen reached a maximum 90




minutes after the injection, returning to control values after three




hours.  Stimulation of gluconeogenesis was also demonstrated in another




experiment using intraperitoneal injection of propanediol.  Quinolinic




acid, a weak inhibitor of phosphoenolpyruvatecarboxykinase (the first




enzymatic step of gluconeogenesis from citric acid cycle intermediates).,




markedly inhibited gluconeogenesis resulting from propanediol adminis-




tration.  According to the authors, this is consistent with the sequence




of glucose formation from 1,2 -propanediol via phosphoenolpyruvatecar-




boxykinase .




     The glucogenic effect of 1,2 -propanediol has also been confirmed




in fasted cows given 150-500 g intraruminally (Voigt and Piatkowski,




1973; Giesecke et al. , 1975).




     These data show that glycogen formation is enhanced by 1,2-pro-




panediol.  Thus, this diol can be used as a synthetic nutrient source.







               3)  Acute Toxicity




                    a)  Oral Administration




                      «   i)  Lethal Dose Values
     For 1,2-propanediol, the LDsg averages 14.3-24.8 g/kg for mice,




21.8-29.0 g/kg for rats and 18.35-19.6 g/kg for guinea pigs; these data




appear in Table 37.  For rabbits, the minimum lethal dose was 20 g/kg




(Braun and Cartland, 1936).





                                    125

-------
                                       Table  37
Oral Lethal Dose Values for 1,2-Propanediol
Species /Strain
mo use /White
mouse/NR
M mouse/NR
rat/Wistar
rat/NR
rat/NR
guinea pig/NR
guinea pig/NR
Sex/ No.
NR/4 per
dose
M&F/10-20
per dose
NR
M/10 per
dose
NR/25
M&F 19-10
per dose
M&F/10
per dose
M&F/10
per dose

Reported value
22.0 cm3/kg
23.9 ml/kga
13.79 cm3/kgb
26.38 g/kg
28 ml/kgb
21.0 ml/kga
18.9 ml/kga
18.35 g/kg
LD50
Deviation
—
ran ge ;
22.8-25.1

95% C.L. =
24.50-28.39
—
range :
19.2-23.0
range:
17.2-20.7
95% C.L. =
16.94-19.87

Converted
value
(g/kg)
22.8
24.8
14.3
26.38
29.0
21.8
19.6
18.35
Reference
Latven and Mo lit or,
1939
Laug et al., 1939
Fischer et al. , 1949
Smyth et al. , 1941
Thomas et al. , 1949
Laug et al., 1939
Laug et al., 1939
Smyth et al., 1941
NR = not reported.

a
 Animals fasted overnight;  observed  for  five  days.
      after 24 hours.

-------
                         ii)  Signs



     In white mice, the minimum symptomatic dose for 1,2-propanediol was


8.3 g/kg (Latven and Molitor, 1939).  In Wistar rats, a  dose of  10


g/kg was without effect (Litton Bionetics, 1974).  Large oral doses


of the glycol to mice, rats, and guinea pigs resulted in loss of


equilibrium, marked depression, analgesia, coma, and finally, death


after a prolonged moribund period (Laug et al., 1939).  Gross observa-


tion revealed hemorrhagic areas in the small intestine.  In the kidney,


microscopic changes included vacuolar degeneration of the cytoplasm


and nuclear pyknosis.  The liver showed slight congestion and hypere-


mia with no fatty changes.


     Gastric administration of 1.5, 2.0, or 10.0 ml/kg (1.6,  2.1, or


10.4 g/kg) 1,2-propanediol to three dogs of unreported breed and sex


in divided doses (6, 4, or 2 doses, respectively) was without effect


(Hanzlik et al., 1939a).


     Three horses (sex and weight not reported) given 0.5-1 gallon


of 1,2-propanediol via a stomach tube developed ataxia and depression


within 15-30 minutes (Myers and Usenik, 1969).  Recovery occurred


within three days without treatment.  A 454 kg gelding was fed two


gallons of the glycol.  Within 15 minutes it became ataxic.  It be-


came recumbent within three hours and remained so until death three


days later.  Initially, diarrhea and abnormal peristalsis were noted.
                     «

At necropsy, the following was observed:  moderate sloughing of the


gastric mucosa; full and dilated stomach, although the gelding had not


eaten in three days; moderate to severe inflammation of the  intestinal


tract; congestion of large colon with ecchymotic hemorrhages on  the
                                   127

-------
serosa of small intestine.  Also,  the kidney was congested and the




brain edematous.   Degenerative changes of the liver cells were seen




microscopically -







                         iii)   Behavioral Effects





     Isgrig and Ayres (1968) evaluated the behavioral effects of 1,2-




propanediol using voluntary activity and equilibrium measures in male




Sprague-Dawley rats.  A hunger-activity cycle was imposed seven days




before testing, and continued for the duration of the experiment;




rats were given access to food for only three hours a day.  Four rats




were given 11.43 ml/kg (11.84 g/kg) 1,2-propanediol by stomach intuba-




tion and the percentage of voluntary activity during the next three




hours was recorded.  Activity in those receiving 1,2-propanediol was during




the second and third hour compared to those given an isocaloric, iso-




volumetric dose of sucrose.







                    b)  Dermal, Ocular, and Ear Application





     USP 1,2-propanediol was a mild irritant to the skin of New Zealand




white rabbits  (2.5-3.9 kg).  In three replicate modified Draize tests,




the glycol was applied to an occluded area of shaved dorsal skin of




four rabbits.  After 24 hours, an average score of 0.39 was obtained,




corresponding to barely perceptible erythema or edema (Phillips et al. ,




1972) .




     Instillation of 0.5 ml 1,2-propanediol into rabbits' eyes caused




moderate edema and hyperemia after 24 hours (Latven and Molitar,




1939) .




     1,2-Propanediol adversely affected hearing in guinea pigs
                                   128

-------
(250-400 g) (Morizono and Johnstone, 1975).  Two pinholes were made




in the tympanic bullae and propylene glycol at concentrations ranging




from 10-100% was instilled into the middle ear cavity;  controls were




similarly treated with Ringer's solution.  The duration of exposure




was 1, 2, or 6 days.  Hearing was monitored by cochlear microphonic




responses.  Effects on hearing loss were variable at concentrations




of 10 or 20%, Ci-e., hearing loss occurred in one animal when the




middle ear was exposed to 10% for six days, but not in  another animal




similarly exposed to 20%).  Concentrations of 50 or 100% 1,2-propane-




diol caused deafness in all six animals tested after one day's exposure.




1,2-Propanediol is used as a vehicle in some ear drop preparations;




the authors caution against its use in preparations instilled in the




middle ear cavity.







                    c)  Inhalation Exposure





     Inhalation of 10% 1,2-propanediol aerosol for 20 or 120 minutes




produced ultrastructural changes in the tracheal epithelium of six




rabbits (2,000-3,000 g) (Konradova et al., 1978).  In particular,




mucous secretion by goblet cells was increased; degeneration and




sloughing off of these cells was also observed.  Two hours after inhala-




tion, no differentiation of new goblet cells was noted.  The method




used to generate the aerosol was not specified; it is not clear what




the level of 10% propanediol aerosol corresponds to.
                                   129

-------
                    d)   Parenteral Administration

                         i)   Lethal Dose Values


     The lethal dose for 1,2-propanediol administered by  injection is

listed in Table 38.   1,2-Propanediol is  more toxic via this  route than

by oral administration.   The LDso  for mice averages 6.6-8.3  g/kg i.v.,

9.7-17.48 g/kg i.p.  and 19.2 g/kg  s.c.  In rats,  the LD50 averages

6.4-6.8 g/kg i.v., 13.5  g/kg i.p., 22.5-29.0 g/kg s.c., and  14-20.7

g/kg i.m.  In rabbits,  the LDso  was 6.5  g/kg.

     Brown and Conine (1971) found no difference  in toxicity to mice

of the optical isomers  of propylene glycol.  For  1,2-propanediol

(dl-propylene glycol).,  the LDso  was 17.48 g/kg;  for d-propylene glycol,

the LDso was 17.41 g/kg and for 1-propylene glycol, 16.61 g/kg.



                         ii)  S igns


     Giri et al. CL970)  reported a maximum nonlethal dose of 10 ml/kg

(10.4 g/kg) i.p. in male 50-60 g Sprague-Dawley  rats.  This  dose of

1,2-propanediol caused mild central nervous system depression lasting

for two or three hours.   The minimum lethal dose  was 15 ml/kg (15.5

g/kg) and the LDioo  was between 25-30 ml/kg (25.9-31.0 g/kg).  Death

resulted from severe CNS depression and respiratory failure.

     Vasquez et al. (1977) reported that 1,2-propanediol produces am-

nesia in mice when administered prior to a training experience.  Fifty
                     C
day old HA/ICR male mice were injected i.v. with 20, 40,  or  60%

aqueous propanediol (2.08, 4.16, or 6.24 g/kg);  controls received saline,

Treatment was given either 30 minutes before or up to three  hours after

training in a 1-trial inhibitory  (passive) avoidance task.  Although
                                   130

-------
                                            Table 38
Parenteral Lethal Dose Values for 1,2-Propanediol
Species/Strain
mouse/White
mouse/SPF NMR I
mouse /NR
mouse / Carworth
Farm
mouse/NR
mouse/NR
mo use /White
Sex/No. Route
NR/4 i.v.
per dose
M&F/10 i.v.
per dose
NR/10 i.p.
per dose
F/NR i.p.
NR/NR i.p.
F/60 i.p.
NR/4 s.c.
per dose
LD50
„ , Converted
, Deviation value
value . ., .
(g/kg)
8.0 cm3/kg 8.3
6.4 ml/kga 95% C.L. = 6.6
6.1-6.9 ml/kg
10.96 ml/kgb 95% C.L. = 11.4
9,715-13,380
rag/kg
127.07 milli- -- 9.7
moles/kg
17.48 g/kg — 17.48
9.36 ml/kg S.E. = 1.20 9.7
18.5 cm3/kg — 19.2


Reference
Latven and
1939
Bartsch et
Molitor,
al., 1976
Davis & Jenner, 1959
Karel et al
., 1947
Brown and Conine,
1971
Karel et al
Latven and
1939
., 1947
Molitor,
rat/White
NR/40
i.v.      6.8 g/kg
6.8
Weatherby and Haag,
1938

-------
                                 Table 38 (Continued)
Species/ Strain
rat/Sprague
Dawley
rat/Sprague
Dawley
rat/NR
rat/NR
rat /White
rat /White
rat/NR
rabbit /NR
Sex/No . Route
" M&F/10 i.v.
per dose
M&F/10 i.p.
per dose
NR/25 i.p.
NR/25 s.c.
NR/15 s.c.
NR/12 i.m.
NR/25 i.m.
NR/18 i.v.

Reported
value
6.2 ml/kga
13.0 ml/kga
13 ml/kgc
28 ml/kg0
22.5 g/kg
14 g/kg
20 ml/kgc
6.5 g/kg
LD50
Converted
Deviation value
Cg/kg)
95% C.L. = 6.4
5.2-7.4
95% C.L. = 13.5
10.2-16.5
13.5
29.0
22.5
14
20.7
6.5
Reference
Bartsch et al
Bartsch et al
Thomas et al.
Thomas et al.
Weatherby and
1938
Weatherby and
1938
Thomas e t al .
Weatherby and
1938

.,1976
. , 1976
, 1949
, 1949
Haag,
Haag,
, 1949
Haag,
NR = Not reported
aLDso determined after 7 days.
 Based on cumulative mortality during a 28-day post-injection period,
°LDso determined after 24 hours.

-------
more pronounced at 60%, treatment at all levels prior to training signifi-




cantly impaired retention performance.  In general, post-trial treat-




ments were ineffective.  According to the authors, the amnestic effect




is likely due to central nervous system depression.




     Intraperitoneal injection of up to 4 ml/kg (4.1 g/kg)  of 1,2-




porpanediol had no narcotic effect in four Wistar rats, two Mongrel




dogs, or two rabbits (Eichhaum and Yasaka, 1976).  The anesthetic




dose was 21.2 ml/kg  (22.0 g/kg). in dogs (Hanzlik et al., 1939a) .




     Intramuscular injection of doses greater than 7.4 g/kg to white




rats resulted in increased respiratory rate, loss of equilibrium, de-




pression, coma, and death (Seidenfeld and Hanzlik, 1932).




     Injection of 0.3 ml of undiluted 1,2-propanediol into the sciatic




nerve of 44 anesthetized adult albino rats caused complete paralysis




in 28 and moderate paralysis in 28 rats CChino et al., 1974).  Histo-




logical changes included diffuse necrotic lesions four days after in-




jection.  Regeneration was noted two weeks following injection when




small axons, and an increased amount of perineural connective tissue




were observed.  Intramuscular injection of 0.1 ml of undiluted pro-




panediol produced localized necrotic lesions.  The authors suggest




from these findings that 1,2-propanediol might be useful for chemical




neurolysis as a means for control of spasticity in patients with cen-




tral nervous system disease.




     Eichbaum and Yasaka (1976) reported that 0.2-0.3 ml/kg  (0.2-0.3




g/kg) 1,2-propanediol (.70% solution) had antiarrhythmic-antifibrilla-




tory effects on the heart when injected intravenously into mongrel




dogs  ( 8-15 kg), and male Wistar rats (400-480 g) with spontaneous or




drug-induced arrhythmias.
                                   133

-------
                         iii)   Hematology and Hemodynamics





     Intravenous injection of 4 ml/kg (4.1 g/kg)  of USP 1,2-propane-




diol (12.5, 25, or 50% in saline)  to 2-3 kg Himalayan rabbits had no




effect on red blood cell count, white blood cell  count, hemoglobin




concentration, or packed cell volume (Brittain and D'Arcy, 1962) .




However, there was a dose-dependent increase in the number of circulat-




ing polymorphs and a dose-dependent decrease in the number of lympho-




cytes.  The number of circulating monocytes, eosinophiles, and baso-




philes remained unchanged.  At the highest concentration of the glycol




administered, there was a marked decrease in blood clotting time and




an increase in plat let count.




     Hemolysis was observed following i.v. administration of sublethal




doses of 1,2-propanediol to rats (molar concentrations 0.278-0.167)




(Weatherby and Haag, 1938).  Lehman and Newman (1937) reported that




if propanediol dilutions (up to 30% propylene glycol) were prepared




with 0.9% NaCl, no hemolysis occurred.




     Two of seven cats given 2 ml/kg (2.1 g/kg) 1,2-propanediol by




intramuscular injection showed very low serum calcium levels within




24 hours.  Four of the cats showed crystals in the convoluted tubules.




However, in six cats given 5 ml/kg (5.2 g/kg) by  i.m. injection,  no




changes in serum calcium levels were detected (Van Winkle and Newman,




1941).




     MacCannell (1969) investigated the effect of 1,2-propanediol




and ethylene glycol on hemolysis or hemodynamic changes in 43 dogs;




this study is discussed in more detail in section II-A-2-C-3).  Both




compounds were infused i.v. for 2-5 minutes at a rate of 20 mg/kg/min
                                   134

-------
or 150-250 mg/kg/min.  At either rate, cardiac output and superior




mesenteric blood flow were increased.  Renal blood flow was markedly




decreased at the higher rate.  Direct injection of 25-50% glycol into




the renal artery resulted in decreased blood flow through this ar-




tery; injection into the superior mesenteric artery increased flow




through this vessel.







               4)  Subacute Toxicity




                    a)  Oral Administration





     Groups of 15 male and 15 female Charles River rats were fed




diets containing 0 (control) or 50,000 ppm (5%) 1,2-propanediol for




15 weeks  (Gaunt et al., 1972).  There were no significant differences




between the control and treated rats for the following:  analyses of




serum and urine; hematological indices; organ weights; and gross




examination, of organs at autopsy.  No data are presented for these




parameters.




     When 1,2-propanediol was fed at levels higher than 30% in the




diet,  weanling rats lost weight and died in a few days (Whitlock




et al., 1944).  Older ("half grown") animals were able to tolerate




higher levels (exact level not given).




     Hyperglycemia resulted in male Wistar rats given 5 or 10% 1,2-




propanediol orally for five weeks (Vaille et al., 1971).




     Daily doses of 1-8 ml/kg (.1.0-8.3 g/kg) were administered by




gavage to 11 rabbits (1.6-2.6 kg) over 50 days (Braun and Courtland,




1936).  Body weight gain was comparable to that of controls.  No




gross pathology was attributed to glycol administration.
                                   135

-------
     Dean and Stock (1974)  presented evidence that 1,2-propanediol




is not an inert solvent.  Administration of 2-6 ml/kg (2.1-6.2 g/kg)




i.p. twice a day for three days to male Wistar rats (170-230 g)




caused a significant elevation of the rate of in vitro hepatic mi-




crosomal metabolism of aniline and p-nitroanisole; also,  there was a




significant decrease in the rate of aminopyrine demethylation but no




change in p-nitrobenzoic acid metabolic rate.  When 1,2-propanediol




and phenobarbital were administered concurrently for three days,  an




additive response was observed in the rate of metabolism of aniline




and p-nitroanisole.  No changes in cytochrome 450 were observed.




In vivo, pretreatment with 4 ml/kg i.p. twice daily for three days




prior to injection of 125 mg/kg hexobarbital resulted in increased




hexobarfaital sleeping times compared to rats receiving hexobarbital




only; this result suggests inhibition of hexobarbital metabolism by




propanediol.  Similarly, administration of 120 mg/kg zoxazolamine




after propanediol pretreatment resulted in increased paralysis times




which was likely due to a reduction in the rate of metabolism of




zoxazolamine.




     Zaroslinski et al.  (1971), however, found no change in hexo-




barbital sleeping time after administration of 4 ml/kg (4.1 g/kg)




1,2-propanediol daily for four days to CF-1 mice.







               5)  Chronic Toxicity




                    a)  Oral Administration




                         i.  Rats




                              i)  Weight Gain





     Weanling male rats (initially 55 g) were given 1,2-propanediol
                                   136

-------
in the drinking water in concentrations of 0, 0.1, 0.3, 1.0,  or 10.0%




for 100 days.  The weight gain of rats receiving the highest  level




was depressed during the first ten days.  All rats thereafter showed




rate of weight gain comparable to controls.  No pathological  changes




were observed which could be attributed to glycol treatment (Weather-




by and Haag, 1938).




     Incorporation of 1-10% 1,2-propanediol in the drinking water




of 20 white  rats  (50 g) for 140 days had no effect on body weight




or growth compared to no-treatment controls (Seidenfeld and Hanzlik,




1932).




     In a paired  feeding experiment, six rats (strain and sex not




given) received 1,2-propanediol by stomach tube in a volume equal to




12.8% of the food intake of the previous day; six control rats were




given an equal quantity of water by stomach tube  (Hanzlik et  al.,




1939a).  Experimental rats showed a higher rate of weight gain during




the 180 day  experiment.




     Groups  of 30 male and 30 female Charles River CD rats were fed




for two years diets which contained 6,250, 12,500, 25,000, or 50,000




ppm 1,2-propanediol (0.625, 1.25, 2.5, or 5%, respectively).   At




the four dietary levels, the mean daily intake of the glycol was 0.2,




0.4, 0.9, and 1.7 g/kg in males and 0.3, 0.5, 1.0, and 2.1 g/kg in




females.  There were no statistically significant differences between




control and treated rats in survival, food consumption, or body




weight gain  (Gaunt et al., 1972).
                                   137

-------
                              ii)   Organ Effects





     One of three rats  receiving for five months a diet in which




1,2-propanediol replaced half of the carbohydrates, showed moderate




degeneration of the liver and of the convoluted tubules of the kidney




(Hanzlik et al., 1939a).




     Compared to controls, no pathological changes were observed in




the kidney, heart, spleen, and liver of 20 white rats which received




1-10% 1,2-propanediol in the drinking water for 140 days (Seidenfeld




and Hanzlik, 1932).




     Slight liver damage was observed in 20 albino rats receiving




2.45 or 4.9% 1,2-propanediol in the diet for two years.  No other




pathological changes were attributed to the glycol (Morris et al.,




1942) .




     No differences in absolute or relative organ weights were noted




among Charles River rats receiving 0-50,000 ppm (.0-5%)  1,2-pro-




panediol in the diet for two years (Gaunt et al., 1972).  There was




a wide spectrum of histological abnormalities,  especially in the




kidney, liver, and lung.  These occurred with equal frequency among




control and treated rats and were consistent with those expected in




aging rats.  As discussed further in Section B-l-b-6, the incidence




of neoplasms was similar among control and propylene glycol treated




animals.








                              iii)  Eematology and Urinalyses





     No hematological differences were found among Charles River rats




fed diets containing 0 to 50,000 ppm (0-5%) 1,2-propanediol for two
                                   138

-------
years (Gaunt et al., 1972).  These parameters were measured at weeks

13, 21, 52, and 80 in eight rats of each sex:  hemoglobin content,

packed cell volume, erythrocyte, and leucocyte counts,  and differ-

ential cell counts.  Reticulocyte counts were made at week 52, 54,

and 80 and hemoglobin counts were made at week 104.

     No significant difference was noted in urinary cell excretion

or renal concentration tests among Charles River rats receiving dietary 0,

25,000, or 50,000 ppm CO, 2.5, or 5.0%, respectively) 1,2-propanediol

for one year  (Gaunt et al., 1972).  At weeks 13, 30, and 52, the fol-

lowing measurements were taken on 6-10 rats of each sex:  specific

gravity and volume of urine produced during a six-hour  period of water

deprivation,  during a two-hour period following a water load, and during

a  four-hour period beginning 16 hours after a water load.  In addition,

urinary cell  counts were taken on the six-hour samples.
                         11.  Dogs


                              i)  Weight Gain


     The general health and weight of four female dogs (breed not


 given,  6.2-21 kg) were unaffected by oral ingestion of 5% aqueous


 1,2-propanediol for 5-9 months; the average daily intake was 5.1


 ml/kg (5.3 g/kg).

     In a two year feeding study, groups of five male and five female

                     «
 beagle  dogs  (.initially 10-14 months old), received 5.0 or 2.0 g 1,2-


 propanediol/kg/day  (Weil et al., 1971).  Isocaloric controls received


 6.35 or 2.54 g dextrose/kg; another control group received no treat-


 ment.   One dog each died in experimental and control groups.  Body
                                    139

-------
weight gain was comparable in dogs receiving either the experimental




diet or no treatment.  Isocaloric control males showed a significantly




greater increase in weight during the first six months compared to




no treatment controls.  The authors suggest that although the pro-




pylene glycol and dextrose diets were isocaloric, the calories in




the diet were not necessarily equally available for conversion to




adipose tissue.  This may explain differences in weight gain.




     Water consumption in dogs treated with 1,2-propanediol was




reduced during the first .year of the study.  Accompanying this was




a low specific gravity of some urine samples and a discrepancy be-




tween water intake and urine volumes.  According to Weil et al. (1971),




these factors are accounted for by the generation of water during




oxidation.  One gram of 1,2-propanediol yields 0.95 g water and 1 g




dextrose yields 0.60 g water when oxidized in the body.







                             ii)  Organ Effects





     Four female dogs (breed not reported; 6.2-21 kg)  were given




1,2-propanediol for 5-9 months and received no other fluid; the




average daily intake was 5.1 ml/kg (Van Winkle and Newman, 1941).




At irregular intervals during the experiment, tests were made of




kidney function (phenolsulfonphthalein excretion) and liver function




(rose bengal in blood; galactose and uric acid in urine).  Compared




to pre-exposure levels, no effect on these tests occurred with glycol




treatment.  Histological examination revealed no pathological changes




in the livers or kidneys.  In four male dogs drinking about 4.5 ml/kg




(4.7 g/kg) daily for 5-6 months, the only change in renal function




noted was elevated blood urea levels.
                                    140

-------
     No differences were measured in relative liver or kidney weights

in dogs receiving 5 or 2 g 1,2-propanediol/kg/day for two  years,

compared to dextrose or untreated controls (Weil et al.,  1971)..   Or-

gans were examined grossly and micropathologically.  Numerous sporadic

lesions were observed in dogs receiving the glycol or dextrose which

were comparable to those found in untreated controls.  No  findings

were considered to be the result of treatment.



                           iii)  Hematology and Urinalyses


     In dogs fed 5.0 or 2.0 g 1,2-propanediol for two years,  Weil

et al. (1971) measured no changes in the following parameters compared

to dextrose or no treatment controls:  differential leucocyte counts,

erythrocyte fragility, urinary pH and micro-examination, alkaline

phosphatase, bromsulphthalein retention, liver glycogen,  blood glu-

cose, total liver lipids, metabolic rate of liver slices  using pro-

pylene glycol, or activities of serum glutamic-oxalacetic and glu-

tamic-pyruvic transaminases.

     In dogs receiving 5 g/kg (but not 2 g/kg) 1,2-propanediol an

effect on erythrocytes was apparent when compared to the  nutritionally


equivalent dextrose group (6.35 g dextrose/kg).  Hemoglobin,  hema-

tocrit and total erythrocyte count were slightly decreased; also,

anisocytosis, poikilocytes, and reticulocytes were increased.  The

                     «
foregoing changes indicate some erythrocyte destruction with replace-

ment from the bone marrow.  There was no evidence of damage to the


spleen or bone marrow.

     Some changes were not dose related and occurred only sporadically,
                                   141

-------
These included a decreased total leucocyte count in males receiving




the lower dose of the glycol,  and a decreased level of liver trigly-




ceride in females given the lower dose.




     Total bilirubin and urine output were significantly increased




in females receiving 5 g of 1,2-propanediol/kg.







                    b)  Inhalation Exposure




                         i.  Rats





     A group of 20 seven week old white rats (.initially 80-90 g)




were exposed to an atmosphere of 1,2-propanediol aerosol (170-350




mg/m3) for 18 months (Robertson at al., 1947).  Weight gain was




slightly higher in the experimental group than in controls.  Both




groups were in good condition and bred regularly.  At sacrifice




made during various intervals after 3-18 months, no abnormal changes




were observed in the kidney, liver, or spleen.  In experimental




animals, round cells accumulated in the lung after five months' ex-




posure .








                         ii.  Monkeys





     Twenty-nine rhesus monkeys  (2959 g) were exposed for up to 13




months to an atmosphere of 100-220 mg/m3 or 230-350 mg/m3 1,2-pro-




panediol (Robertson et al., 1947).  No pathological effects were




attributable to the exposure; organs were examined grossly and micro-




scopically after 1-13 months' exposure.  Urine concentrating ability,




blood cell counts, and hemoglobin determinations in experimental




animals were comparable to controls.
                                   142

-------
               6)  Special Studies

                    a)  Reproduction

                         i.  Mice


     1,2-Propanediol was evaluated for teratogenicity in albino CD-I

outbred mice (Food and Drug Research. Laboratories, 1973a) .   Groups

of 25-28 females were given oral doses of 16.0, 74.3, 345.0, or 1,600.0

mg/kg on days 6-15 of gestation.  Animals were killed on day 17.

1,2-Propanediol was without effect on the following:  the number of

implantation sites, resorption sites, live and dead fetuses; pup

body weight; presence of abnormalities of fetal soft or skeletal

tissues.



                         ii.  Rats


     Groups of 25-28 female Wistar rats were given daily oral doses

of 16.0, 74.3, 345.0, or 1,600.0 mg/kg of 1,2-propanediol on days

6-15 of gestation  (Jood and Drug Research Laboratory, 1973a) .  On

day 20, the number of implantation sites, resorption sites, and live

and dead fetuses were recorded.  Pups were weighed and the soft and

skeletal tissue were examined.  Propanediol was without effect on

these parameters when compared to sham-treated controls.

     In rats, addition of 30% 1,2-propanediol  to  the basal diet

resulted in low rates of reproductive success  (Whitlock et al., 1944);
                     *
no additional information was given in the paper.



                         iii.  Hamsters


     Groups of 24-27 adult female golden hamsters received oral doses
                                    143

-------
of 15.5, 72.0,  334.5,  or 1,550.0 mg/kg 1,2-propanediol on  days  6-10




of gestation (Food and Drug Research Laboratories,  1973a).   On  day




14, the numbers of implantation sites, resorption sites and live




and dead fetuses were determined.  Fetuses were weighed and examined




for anatomical abnormalities.   No adverse effects were attributed




to glycol administration.







                          iv.   Rabbits





     No teratogenic effect was noted in groups of 15-20 Dutch-belted




rabbits given 12.3, 57.1, 267.0, or 1,230.0 mg/kg 1,2-propanediol




on days 6-18 of gestation (Food and Drug Research Laboratory, 1973a) .




Animals were examined on day 29.  Propanediol was without effect




on the  following parameters:  numbers of corpora lutea, implantation




sites,  resorption sites, live and dead fetuses; urogenital tract;




fetal weight; visceral or skeletal abnormalities of fetuses.







                          v.  Chickens





     McLaughlin et al. (1963)  labeled 1,2-propanediol as "nontoxic"




when injected into the yolk sac of fertile white Leghorn eggs prior




to incubation.  Injection of 0.05 ml undiluted propanediol resulted




in a 95% hatch rate.  Walker (1967) reported no mortality in ten




three-day incubated eggs injected with 0.1 or 0.05 ml propanediol




into the yolk.




     No mortality occurred in five day incubated Light Sussex embryos




after injection with 0.1 ml 1,2-propanediol (Clegg, 1964).  When




five day old incubated eggs were immersed for ten seconds in the
                                   144

-------
glycol, mortality was 70% compared to 50% in eggs immersed in water.




     Gebhardt (1968) injected several glycols, including 1,2-propane-




diol, into the air chamber or yolk sac of white Leghorn chick embryos.




When injected into the air chamber, 0.05 ml 1,2-propanediol resulted




in an incidence of up to 27% micromelia, a condition characterized




by reduction and torsion of the lower limbs and by parrot beak.  No




micromelia or other abnormalities occurred following yolk sac injec-




tion of 0.05 ml.  Injection of a higher dose (0.2 ml) of 1,2-pro-




panediol on day 4 into the yolk sac did not result in micromelia but




did  cause the formation of a large liquid containing cyst in 40% of




the  eggs.




     Granich and Timiras (.1969). reported that when 1,2-propanediol




is injected onto the chorioallantoic membrane, toxicity is low.  How-




ever, injection into the air chamber resulted in high toxicity,




possibly due to damage of the vasculature near the site of injection.




     Walker  (1967), found that when 1,2-propanediol was injected




into in vitro preparations of fresh intact and broken yolks, it




remains at the site of injection.




     1,2-Propanediol had a dose-dependent glycogen-depleting effect




on the embryonic myocardium of 8.5 day-old chick embryos  (Delphia




and  Frierdich, 1973).  Fertile White Leghorn eggs were exposed for




three hours to 0.025, 0.040, 0.050, 0.075, 0.100, or 0.200 ml/egg




propanediol or sterile water (controls).  The myocardial  glycogen




level for pooled controls was 10.80 mg/g.  Significant  (p < 0.01)




myocardial depletion occurred at all but the lowest dose  of pro-




panediol  For treated eggs, myocardial  glycogen ranged  from 8.54




mg/g at O.Q4Q ml to 2.68 at 0.200 ml.
                                   145

-------
                    b)   Carcinogenic!ty



     No carcinogenic potential was  attributable to 1,2-propanediol

                                                                »
in Charles River rats receiving up  to 50,000 ppm (5.0%)  in the diet


for two years (Gaunt et al. ,  1972). .  The incidence of neoplasms is


shown in Table 39.   Among both treated and control rats, there was


a high incidence of mammary  fibroadenomas, pituitary adenomas, and


subcutaneous fibrosarcomas.   According to the authors, the random dis-


tribution of other tumors and the lack of any dose-response rela-


tionship show that the tumors were  unrelated to propylene glycol


administration.


     1,2-Propanediol was not carcinogenic when given by s.c. injec-


tion to 18 male mixed strain rats (200 g) (Umeda, 1957).   The back


of rats was injected with 1  ml once a week for 15 months, after


which time the amount was reduced to 0.5 ml.  The experiment was


terminated at 88 weeks.  Eleven rats survived past 43 weeks.  No


tumor was produced at the site of injection in any rat.   In addition,


internal organs at autopsy appeared normal.  One rat, dying on the


403rd day, had adenoma of the hypophysis, which the author claimed


was of spontaneous origin.


     Baldwin et al.  (1968) used 1,2-propanediol as a vehicle in


testing the carcinogenicity of 4-acetamidostilbene and N-hydroxy-


4-acetamidostilbene.  Twenty control female Wistar rats received
                     $

three injections of the glycol (dose not given) over four weeks.  No


tumors were observed for 60 weeks,  when the experiment was terminated


for all groups.


     Dewhurst et al. (1972)  evaluated the carcinogenic potential
                                   146

-------
                                Table 39
     Incidence of Neoplasms in Charles River Rats Fed 1,2-Propanediol
             in the Diet for Two Years CGaunt et al., 1972)
                                      Dietary Levels3 (%)
Organ	
and                         0       6.25      1.25       2.5       5.0
neoplasm
                              28   27   25   27   27   23   28   24    27
                         MFMFMFMFMF

Kidney

  adeno carcinoma         0010000100

lung

  adenoma                1012010200

brain

  malignant astrocytoma  1000000000

pituitary

  adenoma                29274   13    1   12    14

adrenal

  adenoma                1110122031

pancreas

  islet cell adenoma     0010100010

thyroid

  adenoma                1110000000

  carcinoma          ,0100000000

testis

  interstitial cell      0    —   1    —   0    —   0    —   1
  tumor

ovary

  follicular adenoma     —   0    —   0    —   1    —   0    —    0
                                   147

-------
                         Table 39 (Continued)
                                      Dietary Levels3 (%)
Organ	
and                         0       6.25      1.25       2.5       5.0
neoplasm	
             No. ot rats 26   2g   2?   25   27   2?   23   2g   24   2
	examined	


salivary gland

  malignant tumor        0000000100

subcutaneous tissue

  fibroma                0001000000

  fibrosarcoma           1200102033

  rhabdomyosarcoma       0010000000

mammary gland

  fibroadenoma           2   17    3   16    2   15    1   18    1   13

  adeno carcinoma         1003000000

skin

  squamous cell          0000100000
  carcinoma

lymph tissue

  lymphoma               0010001010

abdomen

  fibrosarcoma           1000000000
    a
     Figures represent the numbers of rats with tumors out of the total.
of several six-substituted benzo(a)pyrene derivatives using 1,2-propanediol
                                   148

-------
as the vehicle.  A group of 20 young female Balb/c control mice re-




ceived 1 mg propanediol by injection.  No control animals developed




tumors over a two year period.




     1,2-Propanediol was not found to be carcinogenic when applied




to the dorsal skin of female Swiss mice (Stenback and Shubik, 1974).




Groups of fifty mice were treated with 10, 50, or 100% propanediol




(in acetone), every day for life.  As shown in Table 40, the overall




tumor incidence in treated mice was 40-52%; in untreated and acetone




controls, the incidence was 42 and 44%, respectively.  These dif-




ferences are not significant.  The distribution of tumor types was




similar among glycol-treated and control mice (Table 40).




     Fujino et al. (1965). evaluated the carcinogenicity of 4-nitro-




quinoline N-oxide in 1,2-propanediol by daily lifetime skin applica-




tion to the labial mucosa of male ddN mice.  Neither the 36 glycol




controls nor the 30 untreated mice developed carcinoma of the oral




mucosa.




     Application of 1,2-propanediol to the soft palate of six female




Sprague-Dawley rats (150 g) three times a week for up to eight months




did not result in any evidence of oral cancer.  Propanediol was used




as a control to evaluate 4-nitrochinoline N-oxide in (Wallenius and




Lekholm, 1973) .







                    c)  Mutagenicity




                         i.  Host Mediated Assay





     1,2-Propanediol was tested for mutagenic response  in a host-




mediated assay using male ICR mice as hosts and using Salmonella
                                    149

-------
                                Table 40
Number and Type of Tumors in Swiss Mice Treated with
(StenbSck and Shubik, 1974)
1 ,2-Propanediol .
',. ** N Acetone
(in acetone)
10% 50% 100% 100%
No. animals 50 50 50 50
% tumor-bearing 52 52 40 44
animals
No . tumo rs :
lumphomas 15 13 9 12
lung adenomas 7 13 7 9
liver hemangiomas 124 2
thymomas 121
skin tumors - 2
1,2-Propanediol
Untreated
150
42
26
17
4
6
3
ovarian cystadeno-
  carcinoma

ovarian fibroma

ovarian hemangioma

subcutaneous
  f ibromas

forestomach
  papilloma

mammary adeno-
  carcinomas

others
1

1

2
2

2

1
                                   150

-------
typhimurium (strains G 46 and TA 1530) and Saccharomyces cerevisiae


(strain D3) as indicator organisms (Litton Bionetics,  1974).   In an


acute study, groups of ten mice received 30, 2,500,  or 5,000  mg/kg


propanediol by intubation, followed by an ±.p. dose  of an indicator


organism.  In a subacute study, mice received five daily doses of


propanediol C30, 2,500, or 5,000 mg/kg). and then were inoculated


with the indicator organism.  The mice were killed three hours after


dosing with the test organism.  The induction of reverse mutation was


determined with Salmonella and mitotic recombination was determined


with Saccharomyces.  1,2-Propanediol caused no significant increases


in mutant frequencies with Salmonella TA 1530.  At the high dose only,


propanediol produced a "weak or questionable positive" result with


Salmonella G 46.  According to Litton Bionetics (1974) results from


Saccharomyces D3 were difficult to interpret.  The yeast showed in-


creased recombinant frequencies at all levels of the glycol except


the acute high dose.  The high dose resulted in a low recombinant


frequency, which may have been due to selective killing of the mu-


tants.



                         ii.  Cytogenetic Assay


     To assess possible chromosomal damage in somatic cells,  1,2-


propanediol was tested in an in vivo cytogenetic assay  (Litton Bio-

                     c
netics, 1974).  Groups of 15 male albino rats (random-bred; 10-12


weeks old) received an acute dose of propanediol at 30, 2,500, or


5,000 mg/kg; ,they were killed after 6, 24, or 48 hours and bone


marrow metaphase chromosomes were examined.  In a subacute study
                                   151

-------
groups of five rats were given five daily doses at each of the levels

before sacrifice.  There were no significant aberrations of the bone

marrow metaphase chromosomes in glycol-treated animals.

     Human embryonic lung cultures (WI-38) were exposed to 1,2-

propanediol at 0.001, 0.01, or 0.1 meg/ml.  There was no significant

aberration in anaphase chromosomes (Litton Bionetics, 1974).



                         iii.  Dominant Lethal Assay


     Litton Bionetics (1974) evaluated 1,2-propanediol for muta-

genicity using the dominant lethal test in 10-12 week old random

bred rats.  Groups of ten males were given 30, 2,500, or 5,000 mg/kg

orally in an acute study (one dose) or in a subacute study (one dose

per day for five days).  After treatment, males were mated to two

females per week for 7-8 weeks.  Females were killed 14 days after

exposure to males and examined for deciduomata (early deaths) , late

fetal deaths and total implantations.  Propanediol exhibited no

significant adverse effects in the dominant lethal test and was

considered non-mutagenic at the dosages employed.



          c.  Aquatic Organisms


     In rainbow trout, the lethal concentration range for 1,2-propane-

diol is between 50,000-100,000 mg/Jl.  At the lower range, the trout
                     e
survived while at the upper range, all succumbed (Lennon, 1968 in

Hann and Jensen 1974) .  Based on this information Harm and Jensen

(1974) rate the aquatic toxicity of this glycol to be "0" (an in-

significant hazard).  They suggest, based on sewage decay rates,

that a BOD problem could exist at sub-toxic concentrations.
                                   152

-------
     The 24-hour median tolerance limit (TL )  of 1,2-propanediol




to brine shrimp (Artemia salina) is greater than 10,000 mg/£;  a




precise value was not determined (Price et al.,  1974).




     Tanaka et al. (.1975) tested the effect of several  solvents in




a brine shrimp (Artemia salina) assay.  Propylene glycol at concen-




trations of 1-5% had no effect on viability of the brine shrimp.








          d.  Plants





     1,2-Propanediol inhibited elongation of oat (Avena sativa)




segments at concentrations of 0.5-3% in indoleacetic acid (but not




in water) (Farr and Norris, 1971).








          e.  In Vitro Studies





     Newman et al. (1940) examined the effect of 1,2-propanediol




on isolated cat liver.  Five livers were perfused with 500-600 mg%




propylene glycol, and three with 100-200 mg%; three control livers




were perfused.  1,2-Propanediol decreased the oxygen consumption




and carbon dioxide production of the liver, but increased the glyco-




gen content of the liver, and the lactic acid content of the blood;




the utilization of dextrose by the liver was also depressed.  The




authors were unclear whether these effects showed an impaired or




beneficial function of this glycol.




     1,2-Propanediol depressed the amplitude and frequency of  con-




tractions of in vitro smooth muscle preparations (Bonnardeaux, 1971) .




Duodenal, rectal, and uterine muscles were obtained from adult female




rats of unspecified strain and exposed to concentrations of the glycol
                                   153

-------
at 2.5-25 Ug/ml.  Amplitude and frequency of contractions were measured.


The amplitude of contractions was decreased at levels greater than


2.5 yg/ml for uterine muscle and at 2.5 Ug/ml or greater for duodenal


and rectal muscle.  The frequency of contraction was reduced to a


lesser extent at higher concentrations (maximally at about 10 ug/ml).


The authors suggest that propanediol may be directly affecting cellu-


lar metabolism.
     2 .  1,3-Propanediol


          a.  Humans



     No studies were located on the toxic effects of 1,3-propanediol


to humans.





          b.  Nonhuman Vertebrates


               1)  Metabolism



     According to Van Winkle (1941), the toxicity of 1,3-propanediol


is attributable to the metabolite malonic acid which forms insoluble


calcium salts and is an enzyme inhibitor.  However,  Williams (1959)


stated that its metabolic fate is not known.  Gessner et al. (1960)


were not able to identify any metabolites in the urine of four Chin-


chilla rabbits given 4 g of 1,3-propanediol orally;  they found no
                     i

unchanged diol or malonic acid and suggested that 1,3-propanediol


is oxidized completely to carbon dioxide.




               2)  Acute Toxicity
                                   154

-------
                    a)  Oral Administration




                         i.  Lethal Doses





     Fischer et al. (1949) reported an LD50 of 6.0 ml/kg (6.4 g/kg)




for mice given oral doses of 1,3-propanediol.




     Van Winkle (1941) administered 1-19 ml/kg (.1.1-20.1 g/kg) of




l>3-propanediol to 132 rats of unidentified strain by gastric intu-




bation.  A dose of 18-19 ml/kg was fatal to all ten rats tested.




Mortality was as follows at other doses:
                 Dose
(ml/kg)
16
15
17
11-14
10
1-9
wo . lestea

6
11
5
6-11
15
51
/<> nortaj..

50
64
40
10-18
47
0
     In three cats of unreported strain given 3 ml/kg (3.2 g/kg) of




1,3-propanediol orally, no signs were observed within 48 hours; on




day three, however, the cats refused to eat and vomited after drink-




ing water, and by day seven, all three cats had died (Van Winkle,




1941).  Other cats receiving 3-15 ml/kg (3.2-15.9 g/kg) died within




16 days.  Starvation was suggested as the cause of the delayed




deaths .
                                   155

-------
                         ii.  Glycogenic Action





     1,3-Propanediol showed no glycogenic action in rats, in contrast




to 1,2-propanediol (see section III-B-1) (Van Winkle, 1941) .  Forty




fasted rats were given oral doses of 3,  5,  10, or 15 ml/kg (3.2-15.9




g/kg) 1,3-propanediol and killed after three hours.  Liver glycogen




content was comparable in treated and control animals.




     Opitz (.1958) confirmed that 1,3-propanediol is without a gly-




cogenic effect.  Male rats CL50-250 g) were given an oral dose of




50 mmol (3.8 g/kg) of 1,3 or 1,2-propanediol.  After three hours,




the liver glycogen was 116 mg% in rats receiving the 1,3-propanediol




but was 779 mg% in those receiving the 1,2-isomer.  The liver glyco-




gen content in untreated controls was 138 mg%.







                    b)  Parenteral Administration





     Intravenous administration of 3 ml/kg (3.2 g/kg). 1,3-propanediol




was not fatal to cats.  In contrast, this dose given orally was




fatal to all cats tested (Van Winkle, 1941)..




     Van Winkle (1941) administered 3-7 ml/kg (3.2-7.4 g/kg) 1,3-propane-




diol by i.v. injection to 19 rabbits.  Mortality was as follows:




                   Dose        No. Rabbits     % Mortality






                                                   100




                                                    60




                                                    40




                                                     0




Based on these data, the 50% fatal dose would be between 4 and 5 ml/kg
(ml/kg)
6-7
5 -
4
3

6
5
5
3
                                   156

-------
(4.2-5.2 g/kg) , comparable to 1,2-propanediol.


     I.M. injection of 3-9 ml/kg (3.2-9.5 g/kg) 1,3-propanediol in


42 white rats resulted in the following mortality (Van Winkle, 1941);
                     Dose
ml/kg
8-9
7
6
3-5
1NO . K.3CS

9
5
5
15
/. jxiorta.

100
80
40
0
The 50% mortality would be between 6-7 ml/kg (6.3-7.4 g/kg) which is


about twice that for 1,2-propanediol.




               3)  Chronic Toxicity


     Van Winkle  (1941) conducted a 15 week feeding study in rats of


unreported strain.  Groups of five rats received 0, 5, or 12% 1,3-


 propanediol in the diet or 5 or 10 ml/kg daily (5.3-10.6 g/kg)


by stomach tube.  Most rats receiving 1,3-propanediol showed reduced


food intake compared to controls (.10-11 g food/day compared to 15


g/day in controls) and, consequently, a reduced rate of growth.


The slowest growth rate occurred in the groups given 5 or 12% in


the diet.  All rats receiving 10 ml/kg by stomach tube died at the

                     i
end of five weeks.  Based on a similar study by Van Winkle (1941),


twice as much 1,2-propanediol is required to produce the same effect




               4)  Reproductive Studies


     Gebhardt (1968) evaluated the toxicity of several glycols to
                                   157

-------
White Leghorn chick embryos.  Injection of 0.05 ml 1,3-propanediol




into the air chamber resulted in 5% mortality when administered on




day 0 of incubation and 95% mortality when administered on day 4.




For day 0 embryos, 81% of those surviving developed micromelia and




for day 4 embryos, all survivors developed micromelia.  Micromelia




was characterized by reduction and torsion of the lower limbs and




frequently parrot beak.  No saline controls developed micromelia,




and mortality was 10% when administered on day 0 and 0% on day 4.




When 0.05 ml 1,3-propanediol was injected on various days of incuba-




tion into the yold sac, the following mortality and malformations




were noted:
Day of
incubation
1
2
3
4
5
6
7
8
9
%
Mortality
11
5
5
11
16
0
16
20
20
% Survivors
with micromelia
88
63
53
65
69
79
56
38
19
                  a!9-20 embryos tested/day.
                                   158

-------
Again, no saline controls developed micromelia; mortality in con-




trols was 10.5% on day 4.




     Van Oostrom and Van Limborgh (1976) further described the effect




of 0.05 ml 1,3-propanediol injected in White Leghorn chicken eggs.




Seven experiments each using 31-34 embryos were designed to test




the influence of the injection-site and of the egg-position on mor-




tality and abnormalities in 1,3-propanediol or saline-treated em-




bryos.  Mortality data for these seven experiments appear in Table




41.  No embryos survived treatment of 1,3-propanediol in the air




chamber when eggs were incubated in a vertical position.




     Gross examination of embryos surviving 1,3-propanediol treat-




ment  (experiments 6 & 7, Table 41). revealed a 60% incidence of a




chondrodystrophy-like condition (micromelia) :  the legs and wings




were shortened, the lower leg and metatarsal area were bent dorsally,




and the beak often was parrot-like.  Untreated or saline-treated




control embryos did not show any abnormalities.  Histological exami-




nation of the tibia of treated embryos revealed an underdevelopment




of the periosteal bone collar, which was replaced posteriorly by




endochondral bone trabeculae.  The abnormalities induced by 1,3-




propanediol were unlike hereditary congenital chondrodystrophy or




chondrodystrophy following insulin or sulfonamide treatment.  The




authors concluded, therefore, that the micromelia induced by 1,3-




propanediol represents a highly specific type of artificial micro-




melia.
                                   159

-------
Table 41
Effect of
Experiment No.
and Treatment
1 . none —
2. Isotonic 0.05
saline
3. 1,3-pro- 0.05
panediol
4 . none
5. iso tonic 0.05
saline
6. 1,3-pro- 0.05
panediol
7. 1,3-propane- 0.05
diol
1,3-Propanediol on Mortality of White Leghorn
(Van Oostrom and Van Limborgh, 1976)
T . ^. . ^ Position of egg
Injection site ... . ,7
during incubation
vertical
ml air chamber vertical

ml air chamber vertical

horizontal
ml air chamber horizontal

ml air chamber horizontal

ml yolk sac horizontal

Chicken Embryos
No .of .. . -i . . /a/\
Mortality (%)
eggs
31 3
32 6

31 100

32 6
33 9

31 3

34 12


-------
                    b)  Rabbits





     In rabbits continuous infusion for more than ten hours of 0.1-0.3




g/kg/hr of 1,2-butanediol resulted in a dimunition of muscle tone.




Convulsions were not observed.  In addition, no histological abnormali-




ties were noted in the liver or kidney following this continuous in-




fusion (Strack et al., I960).




     1,2-Butanediol was not irritating to the skin of rabbits when




applied repeatedly nor was there evidence of absorption of toxic




amounts.  When applied to the eyes of rabbits, undiluted 1,2-butane-




diol was painful, irritating, and injurious but a 10% aqueous solution




was without effect (.unpublished data, Dow Chemical Co. in Rowe, 1963).








                     c)  Dogs





     In anesthetized  dogs, i.v. injection of up to 1 g/kg 1,2-butane-




diol had no effect on blood pressure, heart rate, or respiration




(unpublished data, Dow Chemical Co. in Rowe, 1963).








               3)  Subacute Toxicity





     All female albino rats given 5-30% 1,2-butanediol in the diet




for eight weeks survived (Schliissel, 1954).  At 40%, however, death




occurred within 11-29 days.









          c.  Aquatic Organisms





     No studies were  found on the aquatic toxicity of 1,2-butanediol.




Hann and Jensen (1974) hypothesized that the four butanediols




would be "practically nontoxic" to aquatic life.  They suggest,
                                   163

-------
however, that a BOD problem could exist at sub-toxic concentrations.
     2.  2,3-Butanediol




          a.  Humans





     No studies on the biological effects of 2,3-butanediol were




located in the literature.









          b.  Nonhuman Vertebrates




               1)  Metabolism





     According to Neuberg and Gottschalk (1925)., 2,3-butanediol is




excreted partly with, glucuronic acid in the urine of rabbits.  Gessner




et al. (I960) identified a glucuronide of 2,3-butanediol in the urine




of four Chinchilla rabbits after oral dosing (2. g) .   Neither diacetyl




nor acetoin was detected in the expired air or the urine of rabbits




given  1.2-1.5 g 2,3-butanediol.







               2)  Acute Toxicity





     The acute oral lethal dose value (LDSQ) of 2,3-butanediol averaged




8.9 g/kg in mice of unreported strain (Fischer et al., 1949).




     Based on the following evidence 2,3-butanediol does not appear




to produce narcotic effects.  Marcus et al. (1976) administered 6.7




mmol/kg (.0.6 g/kg), i.v. and 16 mmol/kg (1.4 g/kg),  i.p. to Sprague-




Dawley rats; no changes in EEC tracings or a loss of righting reflex




were noted.  Similarly, Menon et  al. (1973) found no central nervous




effects, akensia or rigidity in albino mice given i.p. injections of




500 mg/kg 2,3-butanediol.




                                    164

-------
or urea resulted in a less negative nitrogen balance,  1,3-butanediol




plus urea had an additive effect.  In subjects receiving 1,3-butane-




diol alone, blood glucose levels were significantly lower.   Taken




together, these data indicate that 1,3-butanediol provided  calories




that substituted for calories which, would otherwise have been ob-




tained from protein sources in the diet.  1,3-Butanediol did not




result in any change in serum proteins, white blood count,  hematocrit,




hemoglobin, transaminase enzymes, ketoacids, electrolytes,  cations,




cholesterol, triglycerides, or fatty acids.




     To study possible effects of 1,3-butanediol on endocrine func-




tion and glucose homeostasis, Tobin et al. (1975) gave 27 adult women




a diet containing 40 g butanediol or 40 g sucrose for ten days.  The




experimental procedure is described in Table 42.  Levels of various




serum parameters were determined (Table 42).  There were no signifi-




cant differences in blood glucose, cholesterol or triglycerides be-




tween women fed sucrose, of butanediol.  Levels of serum insulin and




growth hormone were slightly elevated in subjects receiving 1,3-




butanediol, but the differences were not significant.  No evidence




of toxic reactions to 1,3-butanediol were detected.




     Tobin et al. (1975) then examined the effects of 1,3-butanediol




during glucose loading.  Ten adult male and female volunteers ingested




diets containing sucrose or 1,3-butanediol, accounting for 10% of the




total caloric intake for five days; from days 6-10, the diets were




reversed.  Glucose tolerance tests were performed on days 6 and 12,




following a 10-14 hour fast.  In both groups, there was a normal rise




and fall of blood glucose during the four-hour test.  Serum insulin,
                                   167

-------
                                Table 42
     Various Parameters Measured in the Blood of Volunteers Fed 1,3-
                    Butanediola (Tobin et al., 1975)

cholesterol (mg/lQQ ml).
triglycerides (mg/100 ml).
glucose (mg/100 ml)
insulin CyU/ml)
Diet Adjust-
ment Period
194 ± 7d
87.7 ± 8
80.3 ± 1.0
15.8 ± 1.1
Diet Plus
40 g Su-
crose
161 ± 1
115 ± 13
77.4 ± 0.8
14.5 ± 0.9
Diet Plus
40 g Butane-
diolc
167 ± 8
107 ± 8
77.6 ± 1.1
15.2 ± 1.0
HGH (vg/inl)'
7.54 ± 1.91    2.03 ± 0.58
2.84 ± 0.76
     Total of 27 volunteers.

     Five-day adjustment period to a diet with wheat protein as a
nitrogen source.
    c
     After  five-day adjustment period,  half of volunteers were fed a
sucrose diet and half fed a 1,3-butanediol diet for five days; five
days later, the diets were reversed.

     Mean ± standard error of the mean.

     HGH:  human growth ho rmone.
lactate, pyruvate, free fatty acids, triglycerides, g-hydroxybutyrate,

acetoacetate, and growth hormone were not significantly different

between the 1,3-butanediol and sucrose groups during the course of

the glucose tolerance test.   The authors concluded that the mechanism

of 1,3-butanediol-related lowered blood glucose in humans and rats
                                   168

-------
(refer to section C-3-b) is not explained by differences in  the  cir-




culating levels of insulin or growth, hormone.







               2)  Dermal Toxicity





     Fischer et al. CL949) reported no irritation to the skin of the




arm of human subjects after repeated application of 5 g 1,3-butanediol




over 16 days.




     Schwartz (.1962; unpublished data in Dymsza, 1968) patch tasted




200 subjects with 1,3-butanediol.  Patches remained on for 72 hours;




tests were repeated 14 days later on the same area.  There were a




total of eight adverse reactions to the compound among the 200 sub-




jects.




     Shelanski (1974) evaluated the dermal effects of 1,3-butanediol




in 200 human volunteers using a repeated insult patch test.  A pad




was moistened with a 50% solution in water (.about 0.9 ml), then applied




to the upper arm; the pad remained in place for a 24 hour period.




This procedure was repeated 15 times over several weeks on each volun-




teer.  No adverse reactions occurred in any person after the first three




applications.  After four or more applications, only 2 individuals




showed visible skin injury, which was described as mild.   Thus, the




author was unable to sensitize most volunteers to 1,3-butanediol.









          b.   Nonhumari Vertebrates




               1)   Metabolism





     In 1960, Gessner et al.  suggested that 1,3-butanediol was meta-




bolized through 3-hydroxybutyric acid,  although they were not able to
                                   169

-------
detect metabolites in Chinchilla rabbit urine after oral dosing.  Miller




and Dymsza  (1967) disagreed with Gessner et al.; they found no change




in serum and urine ketone bodies in rats fed 20 or 30% 1,3-butanediol




in the diet for 30 weeks and because of this, concluded that 1,3-




butanediol  is not metabolized via 0-hydroxybutyrate.  However, evi-




dence from  other investigators, notably Mehlman and his colleagues,




strongly suggests that in rats, 1,3-butanediol is metabolized via




g-hydroxybutyrate.  Rosmos et al. (1975) confirmed this metabolic




pathway for 1,3-butanediol fed to pigs and chickens.  Evidence for




this pathway includes:  i) formation of blood ketone bodies in vivo;




ii) formation of urine ketone bodies in vivo;  iii) formation of ketone




bodies by liver slices after prefeeding animals with 1,3-butanediol;




and iv) metabolism of 1,3-butanediol added to rat liver extract.  This




evidence, reviewed in Mehlman et al. (1975) is considered in the sub-




sections which follow for studies in rats.







     i)  Formation of Blood Ketone Bodies





     Mehlman et al.  (.197la) measured the levels of blood acetoacetate,




p-hydroxybutyrate, lactate, and pyruvate in male Sprague-Dawley. rats




(240-260 g) fed a diet containing 25% 1,3-butanediol and 30% fat for




three and seven weeks; controls received a 30% fat diet but no 1,3-




butanediol.  As shown in Table 43,  there were high significant increases




in acetoacetate and B-hydroxybutyrate concentrations.   Also, total




blood ketone bodies increased about threefold, which would indicate




either an increased metabolism of fatty acids or conversion of 1,3-




butanediol to ketone bodies.   Blood lactate levels were increased




significantly at seven weeks.
                                   170

-------
                                Table 43
           Blood Levels of Acetoacetate, 3-Sydroxybutyrate, Pym-
                vate, Lactata, and local Ketones  in Rats
                      Fed l,3-3utanediola  (Mehlman
                             at al., 1971a)
umoles/ml
acetoacetate
& -hy droxyfauty rate
pyruvate
lactate
total ketones
Butanediol
3 wk 7
0.19** 0
0.42* 0
0.17 Q
3.8 5
Q.58* 0
Dietb
wk
.12**
.45**
.12*
.9
.63**
Control
3 wk
0.07
0.16
0.22
2.4
0.18
Dietb
7 wk
0.05
C.13
0.23
6.0
0.22
           i-lale Sprague-Dawley rats fed diet of 25% 1,3-butanediol
      for three and seven weeks.

           Values are estimated from figures in original text; each.
      value, data from six rats.

          *p < 0.05 compared to corresponding control.

         **p < 0.001 compared to corresponding control.
     Mehlman et al.  (1971a) also calculated the ratios of the para-

meters listed in Table 43.  The ratio of 3-hydroxybutyrate to acetate

(an indicator of the oxidation-reduction potential of liver mitochondria).

was significantly increased in rats fad 1,3-butanediol for seven weeks.

This increase reflects an increase in the NADH/NAD-f ratio in mitochondria.
                                   171

-------
The ratio of blood lactate  to pyruvate was increased in rats fed 1,3-

butanediol, showing that the cystolic NADH/NAD+ ratio was increased.

The increase in both ratios indicate that the metabolism takes place

in both cytosol and mitochondria.



     ii)  Formation of Urine Ketone Bodies


     Tobin, Mehlman, and Parker (1972) measured the urine ketone body

concentration in Sprague Dawley rats fed a diet containing 20% 1,3-

butanediol and 30% fat for 34 hours.  The total amount of ketone bodies

excreted by treated rats was almost ten times that of control rats

fed a 30% fat diet.  1,3-Butanediol feeding induced a large increase

in p-hydroxybutyrate concentration (2.57 umoles/ml in treated vs.

0.132 in control), a slight increase in acetoacetate concentration

CO.067 umoles/mi in treated vs. 0.044 in controls} and an increase in

the g-hydroxyhutyrate to acetoacetate ratio.



     iii)  Formation of Ketone Bodiesby Liver Slices in Animals
           Prefed Butanediol


     Mehlman et al. (1971a) investigated the formation of ketone bodies

by rat liver tissue from rats who were fed a diet containing 30% fat,

with or without 25% 1,3-butanediol for three and seven weeks.  Liver

slices from both treatment groups were exposed in vitro to glucose,

1,3-butanediol or both glucose and butanediol.  Liver slices from both

controls or rats prefed 1,3-butanediol converted 1,3-butanediol to

ketone bodies.  When 1,3-butanediol was the substrate,  no metabolites

other than ketone bodies were detected-   In rats  prefed with 1,3-fautanediol,

the rate of metabolism of glucose to lactate and pyruvate was decreased;
                                   172

-------
also, the level of g-hydroxybutyrate and g-hydroxybutyrate/aceto-


acetate ratio was decreased.




     iv)  Metabolism of Butanediol Added to Rat Liver Extract


     When 1,3-butanediol was added in vitro to rat liver perfusate,


the  tissue lactate: pyruvate ratio was  increased when  lactate was used


as substrate, an indication of increased cytosolic NADH/NAD ratio


(Mehlman et al., 1971bl.  According to Mehlman et al.(1975)  this sug-


gests that the cytosol is the site of one or more cofactor-dependent


oxidations of 1,3-butanediol.


     Further studies by Tate, Mehlman, and Tobin (1971). strongly sug-


gest that liver alcohol dehydrogenase is the major Cor only) enzyme


responsible for the initial oxidation of 1,3-butanediol to g-hydroxy-


butyrate.  Briefly, rat liver extracts in vitro showed an NAD-f-de-


pendent oxidation  of both 1,3-butanediol and ethanol similar to that


shown by horse liver alcohol dehydrogenase.  Furthermore, known liver


alcohol dehydrogenase inhibitors  (pyrazole and n-butyraldoxime) re-


sulted in a decrease in g-hydroxybutyrate and, therefore, total ketone


body levels.  The  authors suggested that the initial step in the


ADH-catalyzed oxidation of 1,3-butanediol is the formation of 3-hy-


droxybutanal (.aldol) which is then further oxidized to g-hydroxybutyrate,


     Based on a review of the foregoing literature and other papers,

                     e
Mehlman, Tobin, and Mackerer CL975) suggested the following metabolic


pathway for the oxidation of 1,3-butanediol:
                                   173

-------
                    CH3-CH-CH2-CH2OH
                            Cystoplosmic Alcohol OK
                       °HL NAD'
                            NADH + H*

                    CH3-CH-CH2CHO+H20
                        I
                        OH
  Cystoplosmic Aldehyde DH

«• NAD*
  NADH+H*
                    CKs-CH-CHjCOOH
                        I
                        OH
   Mitochondria! /3-Hydroxybutyrate
   NAD*
   NADH+H*
                     CHs-C-CHjCOOH —•  Peripheral Tissue OH
                         II
                         0
                21   Acute Toxicity

                     a)   Lethal Dose Values


     As shown  in Table  44, the oral LDjg  value for 1,3-butanediol

averages 23.4  g/kg in mice, 22.8-29.6  g/kg in rats, and  11.5 g/kg in

guinea pigs.   By injection, the LD^o  is 16.6 g/kg, s.c.  in mice and

20.2 g/kg,  s.c., in rats.  Inhalation  of 1,3-butanediol  "vapors" (ex-

posure, however, must have been to  the aerosol) for eight hours did

not result  in  mortality to Wistar rats (Smyth et al.,  1951).



                     b)   Signs


     1,3-Butanediol had a low acute toxicity to adult  rats in a series
                      c
of studies  by  Bornmann  (1954a, b, and 1955) .  A definite diuretic ef-

fect in males  was  noted, however  (Bornmann, 1954b) .  The amount of

urine in eight rats given an oral dose of 10.0 ml/kg  (10.1 g/kg)

1,3-butanediol averaged 22-39 ml after 24 hours while  in control rats
                                     174

-------
                                                       Table 44
Ui
Lethal Dose Values for 1,3-Butanediol
Species/Strain Sex/No.
**
mouse/NR NR
mouse/NR NR
rat/NR NR
rat/Wistar M/5 per group
rat/Wlstar M/10 per group
rat/NR NR
guinea pig/NR NR
L&50
Route _ ^ ,
Reported
value
oral3 23.31 cm3/kg
s.c. 16.51 cm3/kg
oral3 29.42 cm3/kg
oralb 22.8 g/kg
oralb'° 18.61 g/kg
s.c.a 20.06 cm3/kg
oralb'd 11.46 g/kg

Converted
value
(g/kg)
23.4
16.6
29.6
22.8
18.61
20.2
11.46
Reference
Fischer et al
Fischer et al
Fischer et al
Smyth et al. ,
Smyth et al. ,
Fischer et al
Smyth et al. ,

. , 1949
. , 1949
. , 1949
1951
1941
. , 1949
1941
              24 hours observation period.




              14 days observation period.




             °Standard deviation of LD50 = 21.8-23.9 g/kg.




             °95% C.L. = 17.43-19.88 g/kg.




             d95% C.L. = 10.29-12.77 g/kg.

-------
given 0.9% sodium chloride the amount of urine was 8-10 ml.   No adverse




effects were noted in the organs  of treated animals 24 hours after




dosing.




     1,3-Butanediol is capable of causing intoxication in male Sprague




Dawley rats at acute oral doses higher than 4 g/kg (Hajchrowicz et al.,




1976).  At 6 g/kg ataxia was  observed, at 8 g/kg,  loss of righting




reflex occurred, and at 10 g/kg,  coma was induced.  No signs of in-




toxication were observed at 4 g/kg; this dose actually ameliorated




withdrawal signs in mice physically dependent on ethanol.




     At large acute doses, 1,3-butanediol acts as  a muscle relaxant




(Sprince et al., 19661.  In Sprague Dawley or Holtzmann rats, a dose




of 500 mg 1,3-butanediol/kg i.p.  Cwhich dose of 1,4-butanediol re-




sulted in anesthesia) produced no anesthetic or behavioral effects.




At 7,000 mg/kg, anesthesia was induced.  The response was different




from that occurring with an anesthetic dose of 1,4-butanediol.  There




was extreme flaccidity and loss of limb and body tone; myoclonic




jerking was not observed.  Recovery from anesthesia was slower than




than observed after 1,4-butanediol administration.  In the EEQ tracing,




there were no periods of spiking, polyphasic bursting, or electrical




silence.




     Infusion of 6 ml/kg  (6.03 g/kg) 1,3-butanediol i.v. into dogs




resulted in central nervous system depression, measured by electro-




en cephalography (Stoewsand and Dymsza, 1967).




     Application of 0.01 ml undiluted 1,3-butanediol to the clipped




belly of an albino rabbit resulted in slight necrosis within 24 hours




(Smyth et al., 1951).  Fischer et al. (1949) reported no irritation
                                   176

-------
after dermal application of 1,3-butanediol to guinea pigs or rats.




     One half ml of a 1% solution of 1,3-butanediol applied to the




eye of rabbits caused no reaction within 24 hours (Smyth et al.,  1951).







                    c)  Gluconeogenesis and Lipogenesis





     The effect of 1,3-butanediol on blood glucose levels depends




on the length of treatment.  In acute experiments, injection of 800




mg (Romsos et al., 1974) or feeding 7.5 g/kg 1,3-butanediol (Parker,




1972) resulted in elevated blood glucose levels in Sprague Dawley




rats.  However, in chronic studies, blood glucose levels were depressed.




Mackerer et al. (1975) correlated decreased blood glucose levels with




increased pancreatic insulin content in Charles River rats given up




to 27% butanediol for 30-31 days.  In another study, lowered blood




glucose levels in Sprague Dawley rats given 18% 1,3-butanediol for 14




days were attributed to a block of gluconeogenesis in the kidney at




the  conversion of 3-phosphoglycerate to glyceraldehyde-3-phosphate




(Mehlman et al., 1970).




     Mehlman et al. (1970) also investigated an enzyme involved in




gluconeogenesis (phosphoenolpyruvate carboxykinase, PEPcK) and an




enzyme involved in lipogenesis (malic enzyme).  In treated animals,




hepatic PEPcK activity increased 43%.   Malic enzyme activity was the




same in liver and epididymal fat in treated and control rats indicating




that 1,3-butanediol did not affect lipogenesis.  Mitochondrial syn-




thesis of glucose from pyruvate and bicarbonate by hepatic pyruvate




carboxylase was increased in treated rats.  However, other liver  and




kidney metabolites were unchanged.




     Mehlman et al. (1970) also reported a decrease in the weight






                                   177

-------
of epididymal fat pads (as a measure of epididymal lipid content) in




treated rats.  This had been reported on earlier by Mehlman et al.




(1966) who related higher free fatty acid levels in the plasma to a




mobilization of lipid from epididymal and adipose tissue.  Parker




(1972) suggested that the mobilization of fat would result in




increased blood levels of ketone bodies.




     Romsos et al. (1974) reported a depressed level of fatty acid




synthesis in Sprague Dawley rats given 18 or 36% 1,3-butanediol in




the diet for 23 days.




     Romsos et al. (.1974) suggested that a shift in cytoplasmic




redox state and an increase in hepatic long-chain acyl CoA levels




are involved in the decrease in hepatic fatty acid synthesis after




feeding 1,3-butanediol.  They postulate that the conversion of 1,3-




butanediol to (J-hydroxybutyrate in the liver shifts the cytoplasmic




redox potential toward a more reduced state, thus reducing the rate




of glycolysis and substrate availability for fatty acid synthesis.




The shift in redox potential would spare fatty acid oxidation, causing




long-chain acyl CoA levels to increase.  The latter are inhibitory




to hepatic fatty acid synthesis.  Thus, the overall effect is a de-




crease in the rate of hepatic fatty acid synthesis.







                    d)  Behavioral Effects





      1,3-Butanediol affects voluntary activity in Sprague-Dawley rats




Clsgrig and Ayres, 1968) .  A hunger-activity cycle was imposed prior




to and during testing, in which rats had free access to food for only




three hours a day.  Rats were given 11.43 ml/kg  (11.49 g/kg) by gavage
                                   178

-------
in replicate experiments.  Voluntary activity in a wheel was recorded




over the next three hours.  In treated rats, activity was significantly




depressed during hours 2-3 compared to controls receiving an isoca-




loric, isovolumetric dose of sucrose.  Similar results were obtained




for 1,2-propanediol (section III-B-1).  Effects on equilibrium were




measured in another experiment using rats previously trained to balance




on a rotating bar.  A dose of 11.43 ml/kg significantly disturbed




equilibrium compared to propanediol or sucrose treated rats; however,




the difference was small.  These findings were confirmed in replicate




experiments by Isgrig and Ayres (.1968).  They suggested that 1,3-




butanediol acts as a CNS depressant or strong muscle relaxant.








               3).  Subacute Toxicity




                    a)  Rats





     Bornmann (1954b) added 1, 2,  5, 10, or 20% 1,3-butanediol in the




drinking water of male rats for 13.5 weeks.  The weight of the kid-




ney of rats receiving the two highest doses (.0.216 g/100 g body weight [bw]




at 10%; 0.330 g/100 g bw at 20%) was greater than in controls (0.188 g/100




g bw); no statistical analysis was performed.




     The incorporation of 1,3-butanediol into the diet often results




in a decrease in body weight gain and food consumption.  Typical




findings were reported by Mehlman et al. (1970) for Sprague Dawley




rats fed a ration containing 18% 1,3-butanediol for two weeks; the




decreased body weight gain after two weeks in treated rats was at-




tributed to a decrease in food consumption.  Fischer et al.  (1949),




however, found no effect on weight gain in rats given orally 10 ml/kg




bw from 1-20% solutions of 1,3-butanediol for 44 days.






                                   179

-------
     Feeding 1,3-butanediol also  results  in a decrease in the weight


of epididymal adipose tissue in rats (Mehlman et al.,  1971).   Stoewsand


et al. (1966) reported this effect when it was added  to the diet at

20% for four weeks, and related this decrease to the  reduced resistance


of rats to severe cold stress (—20°C) .

     In a series of subacute feeding experiments,  Miller and Dymsza


(1967) evaluated the ability of rats to use 1,3-butanediol as a syn-


thetic source of energy-  Rats were fed high fat diets C25%)  in which


carbohydrate was replaced by 1,3-butanediol.  In all  experiments,  a

one week period of adaptation was required for maximum utilization of


1,3-butanediol.  After one week,  butanediol had a caloric value of


about 6 kcal/g.  In an ad libitum study C experiment 1), rats fed 20%


1,3-butanediol for four weeks gained significantly less weight than


those with 5 or 10% 1,3-butanediol in their ration; rats fed 10 or

20% 1,3-butanediol consumed less  food than animals on the other diets.


In all groups receiving 1,3-butanediol C5, 10, and 20%), a high level

of food, protein, and caloric efficiency was maintained.  In a. paired


feeding experiment Cexperiment 2), total food consumption and body

weight gain were reduced^in all animals receiving 20% fautanediol for


three weeks.  However, compared to animals receiving  no 1,3-butanediol,

animals pair fed diets containing 5 or 20% 1,3-butanediol had "es-

sentially similar" body weight gains and food and protein efficiencies; in
                     C
the paired feeding,  the amount of the diet fed was controlled by


the group with the lowest food intake.  In rats force fed 20% 1,3-


butanediol, weight gains were comparable to controls  after a one

week adaptation period.
                                   180

-------
                    b)  Cattle


     Bonner et al. (1974) suggested that small amounts of 1,3-butane-

diol can be used as an energy source in cattle rations.  Feeding a

diet containing 4% butanediol to Holstein cows for up to six weeks

was without effect on rumen pE, volatile fatty acid ratios, blood

glucose, blood ketones, body weight, or feed efficiency-  At 6-8%,

however, blood ketones were  .elevated, the animals were hyperactive,

gained less weight and had a lower feed efficiency.

     Young (1975) reported on studies in which. 4% 1,3-butanediol was

fed to lactating fat-depressed Holsteins; treated cows had a greater

milk fat production than controls.



                    c)  Chickens


     In broiler chickens receiving 5% 1,3-butanediol in feed for four

weeks, no effect on weight gain was noted compared to controls (Daven-

port and Griffith, 1969) .  However, weight gain was reduced when

levels exceeded 5% (10, 15, and 21% tested).  Feed conversions were

poorer  when 10% or more was introduced in the diet.  The authors

suggested feed consumption to have been the major factor in reduced


weight gain.

     Davenport and Griffith (1969) found that at dietary levels less


than 5%, 1,3-butanediol can be substituted for part of the fat in
                     e
broiler ration.  No adverse effects were detected in weight or feed

conversion.
                                    181

-------
               4)   Chronic Toxicity




                    a)   Rats





     Kopf et al.  (1950)  administered to male and female rats (85-216




g) an oral dose of 10 ml/kg (.10.6  mg/kg)  of a 50% solution of 1,3-




butanediol every 3-4 days for 45 to  185 days.  No adverse effects




on growth were noted.  Monthly blood counts revealed no differences




in control or treated rats.  After 185 days, rats were sacrificed




and examined; no abnormalities in endocrine organs,  kidney, bladder,




or liver were found.




     Bornmann (1954a and b; 1955)  administered 1-20% 1,3-butanediol




in the drinking water of rats twice a week for 96 days; no toxic




effects were observed except for diuresis.




     Miller and Dymsza (1967) incorporated 20 or 30% 1,3-butanediol




in the diets of rats for 30 weeks to replace carbohydrates.  The




levels of protein, fat, and carbohydrates were varied.




Treated rats received one of the following diets:
Protein
18
36
18
36
Fat
30
30
30
« 30
Butanediol
20
20
30
30
Carbohydrate
22
0
12
0
 Ten  other diet schedules consisted of no 1,3-butanediol, but varying




 combinations of protein, fat, and carbohydrate.
                                    182

-------
At 30% butanediol, but not at 20%, there was a significant impairment




in utilization of butanediol; for example, weight gains were lower




and food and calorie efficiencies were reduced.




     After 30 weeks, urine, blood, and liver were assayed in tests




which included:  urine and serum ketone bodies, liver glycogen,  and




phosphohexose isomerase, and serum glucose.  When the level of fat




was increased in a diet not containing butanediol, serum and urinary




ketone bodies and also the liver glycogen content were increased.




When 1,3-butanediol was added to the diet, these changes did not occur.




Liver, phosphohexose isomerase (PHI) activity and the serum glucose




level generally decreased with increasing fat in the diet, but when




butanediol was added, PHI activity decreased, and no change occurred




in serum glucose.




     In a 476-day feeding study, 20 rats were given 20% 1,3-butanediol




in a 22% casein and 30% fat semi-synthetic diet; 20 controls received




the same diet without the diol.  Lower body weights were noted in




treated rats.  Survival to 476 days was 65% in treated and 100% in




control rats.  Upon histologic examination, no lesions were detected




which could be attributed to treatment CDymsza and Stoewsand, 1965




in Dymsza, 1968) .




     In a two-year feeding study, groups of 30 male and 30 female




weanling Sprague-Dawley rats received diets containing 1.0, 3.0, or




10% (by weight) 1,3-butanediol (Scala and Paynter, 1967).  No dif-




ferences were recorded in any parameter in treated animals compared




to controls.  Parameters examined included body weight gain,  food




consumption, survival hematology  Cerythrocyte and leucocyte counts,
                                    183

-------
hematocrit, hemoglobin)  and urinalysis (specific gravity,  pH, protein,




sugar, acetone, urobilinogen,  bilirubin, occult blood,  and sediment).




Also, organs were weighed and  examined histologically in animals




sacrificed after one or two years.   Experimental findings  were not




attributed to 1,3-butanediol ingestion.  Findings, which included




chronic inflammatory disease of the lungs,  spleen, and kidneys and




spontaneous neoplasms, occurred in  control  and experimental rats and




were consistent with changes common for rats of this strain and age.








                    b)  Dogs





     Kopf et al. (.1950)  orally administered 2.0 ml/kg (2.01 g/kg)




of an aqueous 50% solution of 1,3-butanediol to two dogs twice a




week for 5-6 months.  Sistological  examination revealed no adverse




effects on the liver, kidney,  bladder, or small intestine in the




treated dogs compared to two control dogs.




     In a chronic feeding study, three adult male beagle dogs were




given a diet containing 20% 1,3-butanediol  and 30% fat and three con-




trols received a basal diet containing 30%  fat (.Stoewsand and Dymsza,




1967; Dymsza and Stoewsand, 1966 _in_ Dymsza, 1968).  Endurance capacity




on treadmills was measured four times over five weeks and was com-




parable in control and treated dogs.  Plasma free fatty acids were




significantly lower in treated dogs during weeks three and five.




When the level of 1,3-butanediol in the feed was raised from 20 to




30%  for a short time, incoordination was observed 1-4 hours later.




After 343 days on the test, the dogs were sacrificed.  No gross or




histologic changes of significance were found in any treated dogs,




except for one case of heart worm.
                                   184

-------
     Groups of four male and four female beagle dogs  each received

0, 0.5, 1.0, or 3.0% 1,3-butanediol (dry weight basis)  in the  basal

diet for two years (Scala and Paynter, 1967).   No toxic effects  were

observed at any treatment level.  Daily or weekly measurements were

made of weight, food consumption, and signs.  At eight  intervals,

samples of blood were taken for determinations of erythrocyte  and

leukocyte counts, sedimentation rate, hematocrit, hemoglobin,  blood

urea nitrogen, and bromosulphalein retention.   Urine  was analyzed

for specific gravity, pH, protein, sugar, acetone, urobilinogen,

bilirubin, and occult blood.  Autopsies and histological examinations

were performed on animals sacrificed at 12 or 24 months.  Both control

and test animals showed chronic nephritis.
               5)  Special Studies

                    a)  Reproduction

                         i.  Rats


     In a meeting abstract Dymsza and Adams (1969)  reported on repro-

ductive studies in parents and two generations of rats given diets
                      <:

containing 20% 1,3-butanediol.  Fertility,  litter size, and number

of pups born alive were not affected by dietary treatment.  The
                                   185

-------
three-generation litter survival to three weeks was 77% compared to




51% in controls.  Body weight  gain on 1,3-butanediol was slower than




that of controls; adult weights decreased with each generation.




     The Food and Drug Research Laboratories,  Inc. (FDRL, 1973b)




carried out a series of reproductive and genetic studies on 1,3-




butanediol.  In a four-generation feeding study, groups of 25 male




and 25 female Wistar rats received 1,3-butanediol in a semi-purified




diet to provide 50, 25, 12.5,  or 0% of the calories based on the




metabolizable energy value for the diol of 6.0 calories per gram'




1,3-butanediol accounted for 24, 10, 5, and 0% of these diets, re-




spectively.  The butanediol partially replaced the starch-dextrose




component of the diet.  The parent generation  (Fg) rats were mated




after four weeks on the 1,3-butanediol diet and then continued on this




diet; rats were subjected to two successive mating cycles.  Repro-




ductive performance for the two litters is summarized in Tables 45




and 46; there was a decrease,  particularly in  the second litter, in




the fertility index of 1,3-butanediol-treated  rats.  Longevity studies




were conducted on rats in the  first litter (FI) ', weight gain was de-




creased in animals receiving 1,3-butanediol.  Hematologic, biochemical,




and urinary tests were within  the normal range in treated rats.  FI




females were subjected to five successive mating cycles; in treated




females, fertility decreased progressively with each cycle.  Survival




of FI rats to 77 weeks was somewhat lower in treated rats, but this




difference was not significant  (survival:  control, 66%; 5% diol, 54%;




10% diol, 54%; 24% diol, 52%).  No gross pathological changes noted




at autopsy were attributed to treatment.
                                   186

-------
Table 45
Summary of Reproduction and Lactation Responses in Reproductive
Study in Rats: Data on Fg Generation, Litter^
Fed 1,3-Butanediol (FDRL, 1973b)
1,3-Butanediol Level
number of matings:
number of dams surviving:
number of pregnancies:
number of litters:
cast alive
alive at four days
alive at 21 days
number of pups:
cast alive
cast dead
number of pups:
alive at four days
culled to at four days
alive at 21 days
number of pups per litter:
cast alive
e
alive at four days
culled to at four days
alive at 21 days
Percent
0
25
25
23

23
22
22

229
18

225
164
160

10.0
10.2
7.5
7.3
5
25
25
21

21
20
20

219
6

198
149
147

10.1
9-7
7.5
7.4
10
25
25
19

19
19
19

196
6

188
126
121

10.3
9.9
6.6
6.4
24
25
25
20

20
16
16

185
6

155
117
112

9
9
7
7
















.3
.7
.3
.0
    187

-------
                          Table  45  (Continued)
1,3-Butanediol Level
mean body weight per pup , g
at four days :
at 21 days:
indexes*
fertility
gestation
viability
lactation
0

9.9
85.0

92
100
98
98
Percent
5

10.0
83.0

84
100
91
99
10

10.0
87.0

76
100
96
96
24

10.2
74.0

80
100
84
96
    *Fertility = percent of matings  resulting  in  pregnancies.

     Gestation = percent of pregnancies  resulting in  litters cast
alive.

     Viability = percent of pups  cast  alive  that  survived  at four  days

     Lactation = percent of pups  alive at  four days that survived  to
weaning at 21 days.
     F£ females were mated twice;  no decrease  in  the  fertility  index

was noted.  Pups of the second litter were subjected  to  teratologic

studies.  1,3-Butanediol had no effect on  maternal  or fetal  survival,

on rate of nidation and/or resorption, or  on fetal  soft  or skeletal

abnormalities.  Offspring from the first litter of  the F2 generation
                                   188

-------
Table 46
Summary of Reproduction and Lactation Responses in Reproductive
Study in Rats: Data on Fg Generation, Litte^
Fed 1,3-Butanediol (FDRL , 1973b)
1,3-Butanediol Level
number of mat ings:
number of dams surviving:
number of pregnancies:
number of litters:
cast alive
alive at 4 days
alive at 21 days
number of pups:
cast alive
cast dead
number of pups:
alive at 4 days
culled to at 4 days
alive at 21 days
number of pups per litter:
cast alive
alive at 4 days
culled to at 4 days
alive at 21 days
0
25
25
20

20
19
19

175
10

164
131
131

8.75
8.63
6.89
6.89
Percent
5
25
25
17

17
15
15

149
14

127
98
93

8.76
8.47
6.53
6.20
10
25
25
13

13
12
12

125
6

119
91
87

9.61
9.92
7.58
7.25
24
25
25
16

16
15
13

130
5

114
87
79

8
7
5
6
















.13
.60
.80
.08
   189

-------
                          Table  46  (Continued)
                                           Percent
1,3-Butanediol Level              Q           5         10         24
mean body weight per pup , g
at 4 days :
at 21 days :
indexes*
fertility
gestation
viability
lactation
10.7
63.2

80.0
100.0
93.7
94.7
10.9
57.4

68.0
100.0
85.2
94.9
9.75
55.6

52.0
100.0
95.2
95.6
10.4
53.4

64.0
100.0
87.7
90.8
    *Fertility = percent of matings  resulting in pregnancies.

     Gestation = percent of pregnancies resulting in litters  cast
alive.

     Viability = percent of pups cast  alive that survived at  4  days.

     Lactation = percent of pups alive at  4 days that survived  to
weaning at 21 days.
were reared and later'mated two times.   No treatment related effects

on reproduction or clinical tests were  detected.



                         ii.   Dogs


     Female beagles received 20% 1,3-butanediol in the feed for at


                                   190

-------
least 60 days (dose about 30-50 g/kg/day) then mated and continued


on the treated diet through parturition  (FDRL, 1973b) .  Five-day-old


pups were examined for soft and skeletal abnormalities; no differences


were found compared to control pups.  Treatment was without effect


on maternal health or reproduction.




                         iii.  Rabbits


     On days 6-18 of gestation, Dutch-belted does received oral doses


of 1.3, 2.7, or 5.4 g/kg of 1,3-butanediol in the drinking water


(FDRL, 1973b) . On day 29 of gestation, the uterine contents were ex-


amined for evidence of abnormal fetal development compared to con-


trols, no significant adverse effects were found among treated rabbits.




                         iv.  Chickens


     Injection of 0.05 ml 1,3-butanediol into the yolk sac of White


Leghorn chick embryos resulted in 12% mortality in 0 day-old embryos


(0% in controls) and 89% mortality in 4 day-old embryos (10% in


controls) (Gebhardt, 1968).  No surviving 0 day-old embryos developed


any malformations, while 10% of the 4 day-old embryos developed asym-


metric micromelia (.reduction and torsion of lower limbs; also, parrot


beak).  When 0.05 ml was injected into the yolk sac of 4 day-old em-


bryos, mortality was 22.1% (.10.5% mortality in controls); no malforma-

                     e
tions were detected.


     Partial immersion of five-day incubated Light Sussex chicken


eggs for ten seconds in l,3~butanediol resulted in 40% mortality and


no abnormalities; for water controls, a 50% mortality was recorded


(Clegg, 1964).


                                   191

-------
                    b)  Mutagenicity

                         i.  Dominant Lethal Test

     Male Wistar rats, receiving 0,  5,  10,  or 24% 1,3-butanediol in

the diet for about 80 days, were mated  with virgin untreated females

over an eight week period;  parents of these males had received the

same level of the diol (.FDRL, 1973b). An average mutagenic index (re-

sorptions/implant sites)  was calculated for the mated females:
„ mutagenic
index
0
5
10
24
5.49
6.05
4.34
3.13
                         ii.   Cytogenetic  Studies

     Bone marrow samples were taken from weanling Wistar rats whose

parents had received 0, 5, 10, or 24% 1,3-butanediol in the diet

(FDRL, 1973b).  Metaphase cells were examined.   The occurrence of

abnormal cells was not higher among treated than control rats (0.7-

3.6% of cells abnormal in all groups).


          c_  Aquatic Organisms

     No data were located on the aquatic toxicity of 1,3-butanediol.

Hann and Jensen (1974) suggested that the aquatic toxicity of this

compound might be rated as a "1," "practically nontoxic."  They sug-

gest a BOD problem could exist at sub-toxic concentrations.
                                   192

-------
     4.  1,4-Butanediol

          a.  Humans

     Reports of adverse effects of 1,4-butanediol to humans are not

available.

           b.   Nonhuman Vertebrates
                1)   Metabolism

      1,4-Butanediol appears  to  be  metabolized via succinic acid.

 Four Chinchilla rabbits  (2-3 kg) were  given a total oral dose of 9 g

 1,4-butanediol.  Seven percent  of  the  dose was  recovered in the urine

 as  succinic acid.   No  unchanged diol was  identified in  the urine

 (Gessner at al., I960}.

      The central nervous system depression induced by 1,4-butanediol,

 discussed in section  b-2)    is mediated  through a metabolite, gamma-

 hydroxybutyrate.  Roth and Giannan (1968) identified this metabolite

 in  blood and brain tissue of rats  1.5  hours after intravenous injec-

 tion of 5.8 meq/kg.

      Maxwell and Roth (.1972) showed that  brain, liver,  kidney, and

 heart can convert 1,4-butanediol  to gamma-hydroxybutyrate.  Tissue

 slices or minces from male Sprague-Dawley rats  were incubated for 30

 minutes in a medium containing  0.94 yc of radio-labeled 1,4-butanediol.

 In  other experiments,  5.8 meq of 1,4-butanediol was injected

 intracisternally or intravenously. In all experiments, levels of

 gamma-hydroxybutyrate-were higher  in  the  liver  than the brain, kidney,

 or  heart.   That the liver is the primary  site for conversion  to  gamma-

 hydroxybutyrate was confirmed in an experiment  on partially hepatec-

 tomized rats; these rats showed a  reduced rate  of formation of  the

 metabolite and also a  resistance  to 1,4-butanediol-induced anesthesia.

      In mice, as in rats, 1,4-butanediol  is probably converted to
                                    193

-------
gamma-hydroxybutyric acid.  Menon et al. (.1973)  compared the effects
of equimolar doses (ip)  of these two compounds in male Swiss mice.
Both compounds caused marked akinesia and rigidity, marked hypothermia
and similar changes in brain monoamine levels CSection 2-b ).  Menon
et al. (1973) suggested that quantitative differences between the com-
pounds might possibly be due to differences in rate of transport.
               2)  Acute Toxicity
                    a)  Lethal Dose Values

     1,4-Butanediol is the most acutely toxic of the butanediol iso-
mers.
     The acute median oral LDgQ value for 1,4-butanediol is 2.1-2.2
g/kg in mice, 1.5-1.78 g/kg in rats, 1.2 g/kg in guinea pigs, and  2.5
g/kg in rabbits.  By i.p. injection, the median LDso is 1.3-1.4 g/kg
in rats (Table 47) .

                    b)  Signs

     Signs of toxicity elicited by 1,4-butanediol are different from
those of the other isomers.  For example, while acute doses of 1,3-
or 2,3-butanediol  CO-6 g/kg i.v. and 1.4 g/kg i.p.) did not cause
changes in EEC tracings or a loss of righting reflex in Sprague-Dawley
rats, 1,4-butanediol (0.3 g/kg i.v. and 0.4 g/kg i.p.) resulted in
high amplitude slow wave EEC tracings within five minutes of adminis-
tration and a loss of the righting reflex within 20 minutes, at which
time the EEC showed spiking and periods of electrical silence (Marcus
et al., 1976).  According to Marcus et al. (.1976) 1,3- and 2,3-butane-
diols may fail to produce a central nervous system effect because they
are too hydrophilic to easily pass the blood-brain barrier.
     Menon et al.  (.1973) also found no central nervous system effects
                                   194

-------
                                                   Table 47
Ln
Lethal Dose Values for 1,4-Butanediol
Species/Strain Sex/No.
mouse/NR NR
mouse/NR NR
rat/Wistar NR
rat/NR NR
rat/Wistar M&F/36
rat/Sprague-Dawley M/NR
guinea pig/NR NR
rabbit /NR NR
LD5Q
„ .. converted
Reported ,
, value
value / /i \
(g/kg)
oral 2,062 mg/kg 2.1
orala 2.14 2.2
cm3 /kg
oral 1,525 mg/kg 1.5
oral 1.78 g/kg 1.78
l.p.b 11.87-11.90 1.1
mmol/kg
i.p. 1,328 mg/kg 1.3
oral 1,200 mg/kg 1.2
oral 2,531 mg/kg 2.5


Knyshova, 1968
Fischer et al., 1949
Knyshova, 1968
Dow Chemical Co., in
Rowe, 1963
Taberner and Pearce, 1973
Zabik et al., 1974
Knyshova, 1968
Knyshova, 1968
                 after 24 hours.




                 after 18 hours.

-------
after 1,3- or 2,3-butanediol administration but did after 1,4-




butanediol.  I.P. injection of 143 or 250 mg/kg of 1,4-butanediol




to male Swiss albino mice resulted initially in reduction in loco-




motor activity and exophthalmos.   Within 10-15 minutes, the animals




assumed a spread-eagle position;  some muscular movement was observed.




This akinesia and rigidity did not occur when 1,3- or 2,3-butanediol




was administered.




     Rats appear to be more susceptible to the central nervous system.




effects of 1,4-butanediol than mice.  Anesthesia, measured as loss




of righting reflex, could not be induced by i.p. injection of up to




1 g 1,4-butanediol/kg to Swiss albino mice.  In contrast, a dose of




350 mg/kg to rats Sprague-Dawley rats caused a loss of the righting




reflex within 24 minutes (Menon et al., 1973).  A dose of 250 mg/kg




i.p. to Sprague-Dawley rats had no effect on righting reflex although




at 50-200 mg/kg, spontaneous motor activity was reduced; at 200 mg/kg,




rotorod performance was significantly affected (Zabik et al., 1974).




Sprince et al. (.1966) reported an optimal anesthetic dose of 500




mg/kg i.p. for Sprague-Dawley rats.  In Wistar rats injection of




sub-lethal doses (<10 mmol/kg) produced an hypnotic state within 20




minutes, accompanied by a loss of the righting reflex; muscle tone




was maintained.   A dose of 500 mg/kg, i.p. resulted in a significant




lowering of body temperature.  Higher doses produced a deeper hypnosis




with a marked bradycardia, analgesia, and labored respiration.  Death




was due to respiratory failure (Taberner and Pearce, 1973).




     The narcotic effect of 1,4-butanediol in rats has been confirmed




by Hinricks et al. (1948) for oral administration.  Oral doses of
                                   196

-------
0.12 and 1.92 ml were fatal within 313 and 137 minutes, respectively,




to 50-100 g rats of unreported sex and strain.  Signs included rapid




narcosis, constriction of the pupils and loss of reflexes; death was




attributed to central nervous system paralysis.




     The central nervous system effects of 1,4-butanediol are probably




mediated through a metabolite, gamma-hydroxybutyrate, as previously




discussed.




     In male Swiss mice, i.p. injection of 250 mg/kg 1,4-butanediol




resulted in an increased level of brain dopamine 0.70% of control




at 20 minutes) and a decreased level of brain norepinephrine (60%




of control at 20 minutes).  Brain serotonin remained unaltered (Menon




et al., 1973) .








                    c)  Dermal Effects





     1,4-Butanediol elicited slight irritation when applied to the




eyes of rabbits; very slight conjunctival irritation and no corneal




injury were noted.  No irritation or absorption of toxic amounts were




induced with repeated application to abraded or intact rabbit skin




(.unpublished data from Dow Chemical in Rowe, 1963).
                3)   Subacute Toxicity





      Fourteen daily doses of 500  or 1,000  mg 1,4-butanediol per kg,




 i.p., were without effect on liver triglycerides in male Sprague-




 Dawley rats (300-350 g) (Zabik et al., 1974).
                                   197

-------
               4)  Reproduction Studies





     Injection of 0.05 ml 1,4-butanediol into the air sac of White




Leghorn chick embryos resulted in 35% mortality in 0 day-old embryos




(10% in control) and 89% in 4 day-old embryos (0% in controls)




(Gebhardt, 1968).  Injection of 0.05 ml into the yolk sac of 4 day-old




embryos resulted in 5.6% mortality, which is less than for the control




(10%) .  No malformations were detected in surviving control or treated




embryos.




     A ten-secoad immersion of five day incubated Light Sussex eggs




in 1,4-butanediol resulted in 40% mortality;, compared to 50% in water




controls  (Clegg, 1964).  No abnormalities were detected.







          c.  Aquatic Organisms





     No information was located on the aquatic toxicity of 1,4-butanediol,




Hann and Jensen (1974) speculated that the four butylene glycol isomers




would be  "practically nontoxic" to aquatic life but  that a BOD problem




could exist at sub-toxic concentrations.
                                   198

-------
               IV-  REGULATIONS. STANDARDS, AND HANDLING



A.  Federal Regulations


     1.  Food and Drug Administration



     The Food and Drug Administration regulates 1,2-propanediol and 1,3-


butanediol as both, direct and indirect food additives and ethylene gly-


col, 1,4-butanediol and butylene glycol (isomer unspecified) as indirect


food additives.  A description of these approved food additive uses


appears in Table 48.  1,2-Propanediol is classed as a generally recog-


nized as safe  CGRAS)  food additive and is approved as an emulsifying


agent, as a general-purpose  food additive or as a component in food


packaging; indirect additive uses include applications in adhesives,


resins, solvents, and coatings.  Ethylene glycol is approved for some


uses in adhesives intended for packing, transporting, or holding food,


as  a component of resins, in defoaming agents and in resins.  1,3-Bu-


tanediol can be  added directly to food as a solvent for natural and


synthetic flavoring substances and indirectly as a component of adhesives,


cellophane, and  sealing gaskets.  1,4-Butanediol is approved as a com-


ponent of adhesives.


     On June 23, 1978, the FDA proposed to set  exposure limits for the


sterilant ethylene oxide and its two major reaction products, ethylene


glycol and ethylene chlorohydrin in certain drug products and medical


devices (Gardner, 1978).  Proposed residue limits for ethylene glycol
                      c

are as follows:
                                     199

-------
                                                       Table 48
                                   FDA Status of the Glycols Used as Food Additives
                                             (21 CFR; Union Carbide, 1978)
        Section
        in
        21 CFR
            Description of Approved Uses
Ethylene
glycol
1,2-
Propane-
diol
1,3-
Butane-
diol
1,4-
Butane-
diol
Butylene gly-
col (isomer
unspeci-
fied)
o
o
.A. Direct   Food Additive

172.850     adjuvant for production of
            lactylated fatty acid esters
            intended for use as emulsi-
            fiers, plasticizers, or sur-
            face-active agents in food

173.220     solvent for natural and syn-
            thetic flavoring substances

182.1666    emulsifying agent or as gen-
            eral-purpose food additive, or
            as a component in food pack-
            aging

B. Indirect Food Additive
                                                                    X
                                                                                X
                                                                    X
        175.105     component of adhesives used        X
                    in articles intended for pack-
                    aging, transporting, or hold-
                    ing food

        175.300     component of various resinous      X
                    and polymeric coatings.  Applied
                    as continuous films or enamels over
                    a metal or other suitable sub-
                    strates.  Coatings serve as func-
                    tional barriers between food and
                    substrates and are intended for re-
                    peated contact with food
                                                            X
                            X
                          X

-------
                                         Table 48 (Continued)
Section
in
21 CFR
Description of Approved Uses
Ethylene
 glycol
1,2-
Propane-
diol
1,3-
Butane-
diol
1,4-
Butane-
diol
Butylene
glycol
(isomer
unspecified)
175.320     component of resinous and polymeric
            coatings for polyolefin films in-
            tended for producing, manufacturing,
            processing, preparing, treating,
            packaging, transporting,  or hold-
            ing food

175.380     component of xylene formaldehyde
            resins condensed with 4,4-isopro-
            pylidene diphenolepichlorohydrin
            epoxy resins

175.390     solvent (removed by water washing)
            use in the preparation of zinc-sili-
            con dioxide matrix coatings that  have
            contact with food surfaces in bulk
            re-usable containers intended for
            storing, handling, and transporta-
            tion of food.

176.170     component of coated or uncoated pa-
            per and paperboard intended for use
            in contact with fatty, aqueous, and
            dry foods

176.180     component of coated or uncoated paper
            and paperboard intended for use in
            contact with dry food
                                         X
               X
                                                    X
                                         X
               X
                                                    X
                                                    X

-------
                                             Table 48 (Continued)
      Section
      in
      21 CFR
            Description of Approved Uses
Ethylene
 glycol
1,2-
Propane-
diol
1,3-
Butane-
diol
1,4-
Butane-
diol
Butylene
glycol
(isomer
unspecified)
O
176.200     component of defoaming agents used
            in preparation and application of
            coatings for paper and paperboard

176.210     used in preparation of esters         X
            from fatty acids and alcohols de-
            rived from fatty tryglycerides and
            marine oils.  Used in formulation
            of defoaming agents employed in
            the manufacture of paperboard and
            paper prior to and during sheet
            forming operations

177.1200    component of base sheet cellophane
            of coatings applied to cellophane
            to impart desired properties

177.1210    component of closure sealing gas-
            kets and overall discs for food
            containers

177.1240    component of 1,4-cyclohexylene di-
            me thy lene terephthalate and 1,4-
            cyclohexylene dimethylene isophtha-
            late copolymers

177.1400    component in base sheet or of coatings
            applied to water-insoluble hydroxyethyl
            cellulose film used for packaging food
                                                                      X
                                                                                                        X
                                                                      X
                                                                      X
                                                                      X
                              X
                              X
                                                                      X

-------
                                           Table 48 (Continued)
Section
in Description of Approved Uses
21 CFR
Ethylene
glycol
1,2-
Propane-
diol
1,3-
Butane-
diol
1,4-
Butane-
diol
Butylene
glycol
(isomer
unspecified)
to
o
CO
177.1630    component of polyethylene tereph-
            thalate film used for packaging,
            transporting, or holding alco-
            holic beverages that do not exceed
            50 per cent alcohol by volume

177.1680    component of polyurethane resins
            used as the food contact surface
            for dry bulk food

177.2420    component of crosslinked polyester
            resins used in articles intended
            for repeated use in contact with  food

177.2600    plasticizer in rubber articles in-
            tended for repeated use.  Limit of
            30 per cent by weight of the rubber
            product

177.2800    adjuvant in the production of tex-
            tiles and textile fibers intended
            for contact with dry food

178.3300    adjuvant employed in use of corro-
            sion inhibitors used for steel or
            tinplate

178.3740    plasticizer in polymeric food pack-
            aging materials
                                                                     X
                                                          X
X
                                                                     X
                                                                     X

-------
                                         Table 48 (Continued)
Section
in
21 CFR
Description of Approved Uses
Ethylene
 glycol
1,2-
Propane^
diol
1,3-
Butane-
diol
1,4-
Butane-
diol
Butylene
glycol
(isomer
unspecified)
178.3910    component of surface lubricants
            used in drawing, stamping,  and
            forming of metallic articles from
            rolled foil or sheet by further
            processing

-------
                                             ppm
                                             Ethylene
                                             glycol
              Drug Product

                ophthalmias (.for topical        60
                  use

                injectables (including          20
                  veterinary intramammary
                  infusions

                intrauterine device Ccon-       10
                  taining a drug)

                surgical scrub sponges         500
                  (containing a drug)

                hard gelatin capsule shells     35

              Medical Device

                implant:

                  small (<10 grams)          5,000

                  medium (10-100 grams).      2,000

                  large (>100 grams)           500

                intrauterine device             10

                intraocular lenses             500

                devices contacting mucosa    5,000

                devices contacting blood       250
                  (ex vivo)

                devices contacting skin      5,000
                 i?
                surgical scrub sponges         500
2.  Environmental Protection Agency


There are no specific EPA regulations governing water or air quality

                               205

-------
with respect to the glycols.  In 40 CFR section 180.1001(c), the EPA




lists propylene glycol as one of many inert (or occasionally active)




materials exempt from tolerances for pesticide formulations applied to




growing crops or to raw  agricultural commodities after harvest.




Propylene  glycol is also exempt from the requirement of a tolerance




when used in formulations applied to animals (40 CFR 180.1001-e).  In




both applications, propylene glycol is used as a solvent or cosolvent.




Recently, EPA has proposed that pesticide applications of ethylene




glycol (involving use as an inert ingredient)  be exempt from tolerances




for pesticide chemicals when used in foliar application to peanut




plants (43 FR 29809, July 11, 1978).










        3.  Occupational  Safety  and  Health Administration





        None of the  glycols  considered in  this  report  are  specifically




   regulated by the  Occupational Safety and Health Administration.







        4.  Department of Transportation





        The  glycols  are not  regulated  by the Department of Transportation




   so therefore do not have  DOT  shipping names, hazard classifications,  or




   warning labels.









   B .  State Regulations





        Agencies in^!5 states were  contacted about state-level  regulations




   of the glycols;  their  responses  are listed  in  Tables 49 to 51.







        1.  Workplace Standards





        State agencies responding indicated that  they  rely on the  standards
                                206

-------
                  Table 49
ns for Glycols Food
Selected States
Contact and Workplace £
in Response to Queries
Food Workplace
Contact Standards
California.
Connecticut
Delaware
Kentucky
Louisiana
Missouri
New Jersey
New York
Ohio
Pennsylvania
South Carolina
Tennessee
Texas
* *
* ft
* *
-
-
-
* *
*
-
_ ft
* *
*
— —
    *Federal standards followed; dashes
indicate no response.
                      207

-------
                           Table 50
      Water Standards for the Glycols in Selected States
                    in Response to Queries
State
                 Standard
California

Connecticut
Delaware

Kentucky

Louisiana

Missouri

New Jersey


New York

Ohio

Pennsylvania

South Carolina


Tennessee


Texas
not specifically controlled

no specific water quality standards or
regulations; subject to case-by-case
technical permit review by Water Com-
pliance and Hazardous Substance Unit

not specifically controlled

no response

case-by-case permit review for discharge

general wa'ter quality standards apply

comply with federal discharge require-
ments under FWPCA

no specific water quality standards

general water quality standards apply

general water quality standards apply

indirectly limited by general water
quality criteria

general standards for toxic substances
and organics apply

measured indirectly as BODs or COD in
individual discharge permits issued
to manufacturers
                              208

-------
                           Table 51
           Air Standards for the Glycols in Selected
                 States in Response to Queries
State
            Standard
California



Connecticut



Delaware



Kentucky

Louisiana

Missouri

New Jersey



New York



Ohio
Pennsylvania
South Carolina

Tennessee

Texas
controlled as "reactive organics" in Cali-
fornia basins in which federal oxidant
standards are exceeded

emission of organic material restricted
to 160 Ib/hr, 800 Ibs/day; State law:
Section 19-508-20f(4)

general air quality standards apply; State
law;  Reg. 1-XXlll of Dept. Nat. Res.  & Env.
Control (for air pollution)

no response

no specific regulations

no specific regulations

general ambient air quality standards  apply;
State law:  NJEPA N.J. Adm. Code Title 7,
Chp. 27

processes, exhaust and/or ventilation  systems
are regulated under State law:  Industrial
Process Air Pollution Control Rule,  part 212

The glycols are considered not photochemi-
cally reactive CNPR). by the Ohio Environ-
mental Protection Agency-  There are no
specific regulations for NPR compounds.  State
law:  OAC 3745-21-01 CO

no weight rate emission limitations standards
for organic compounds apply if storage, load-
ing, water separation or the operation of
pumps and compressors would be involved in
their handling. State law:  Pa. Air Pollution
control Act

no specific regulations

general process emission standards; State
law:  1200-3-7-.07
regulated as volatile carbon compounds, State
law:  Reg. V rule 505.2
                              209

-------
set by the Occupational Safety and Health Administration (OSHA), (Table




49).  As discussed in the previous section, OSHA does not regulate the




title glycols.







     2.   Food Contact





     States queried enforce the regulations of the FDA and the USDA (Table




49).





     3.   Water Quality





     No  specific water quality regulations exist in the states questioned.




Rather,  the glycols are indirectly limited by general water quality




standards (Table 50).







     4.   Air Emissions





     The title glycols are not specifically regulated in the states




queried (Table 51).









C.  Foreign Countries





     Agencies in several foreign countries were contacted about regu-




lations concerning the glycols.







     1.   United Kingdom





     In the United Kingdom, occupational standards exist only for ethy-




lene glycol, which are the following:
                                   210

-------
     particulate:


        threshold limit value (TLV)  — 10 mg/m3


        short-term exposure limit CSTEL)  — 20 mg/m3



     vapor;


        TLV — 100 ppm (260 mg/m3)


        STEL — 125 ppm (325 mg/m3)


     1,2-Propanediol is permitted as a solvent in food under the Sol-


vents in Food Regulations (1967) to the extent set out in the British



Pharmacopoeia (1963).


     Air pollution control for industrial processes is regulated by HM


Alkali  and  Clean Air Inspectorate in England and Wales and by HM In-


dustrial Pollution Inspectorate  in Scotland.  There are currently no


specific air  or water  standards  for the  glycols.


     Ethylene glycol,  1,2-propanediol and 1,3-butanediol are permitted


as humectants in  tobacco according to the Ordinance on Tobacco.   Ethylene



 glycol  and  1,3-butanediol  are  also  permitted  as  solvents in  the produc-


 tion of cigarettes,  cigars,  smoking tobacco,  and snuff.  1,2-Propanediol


 is permitted as a solvent  for  flavorings and  as  a solvent  for anti-oxidants;



 in food,  1,2-propanediol may not exceed  500 mg/kg.




      2.  West Germany



      No workplace standards  exist  for the glycols under consideration.
                     c

      Air  emission standards  for ethylene glycol  appear in  the Air Purity


 Regulations (6  MBI, 1974).   In waste gas,  ethylene glycol  cannot  exceed


 300  mg/m3 when  the  flow rate is 6  kg/hour or  higher.
                                   211

-------
     3.  Japan


     The glycols do not appear in a list of toxic substances which are


controlled under the Air Pollution Control Law (Law No. 97;  1968).


     The Water Pollution Control Law (Law No. 138;  1970}  regulates a


number of industries including the textile industry, synthetic plastic


industry, and organic chemicals industry.  Specific levels of glycols


do not appear to be regulated under this law.




     4.  Canada


     In Canada, occupational health is under provincial rather than


federal jurisdiction.  In general, provincial standards follow ACGIH


and/or OSHA recommendations.


     The Environmental Protection Service (EPS)  in Canada is currently


developing regulations and guidelines to reduce  air and water pollution.


Effluent guidelines are being written for the organic chemicals indus-


try (Anon, 1978b); these might possibly regulate the glycols.


     Ethylene glycol is not permitted as a food  additive.  Use of 1,2-


propanediol and 1,3-butanediol is allowed as a food additive; good


manufacturing practice must be followed.  Glycol esters may be used in

certain pesticide f®naulations as solvents.




D.  Other Standards  — Threshold Limit Value

                       
-------
Limit (STEL) maximum for ethylene glycol be 125 ppm (325 mg/m3)  for vapor




exposure and 20 mg/m3 for particulate exposure.  The ACGIH has not recom-




mended exposure limits for the other glycols.









E.  Handling and Storage Practices




     1.  Handling, Storage and Transport





     The glycols are considered stable, noncorrosive chemicals with high




flash points.  They can be stored in mild steel vessels under ordinary




storage conditions.  Vessels lined with a baked-phenolic resin,  epoxy-




phenolic resin, or vinyl resin may be used for long-term storage or




if trace iron contamination and the development of color would be objection-




able; alternatively, stainless steel or aluminum tanks can be used for




storage, but these are more costly-  For above ground outside storage,




tanks and lines may be heated because glycols become relatively  viscous




at low temperatures.  Excessive temperatures can result in product degra-




dation (Union Carbide, 1978).




     Ethylene glycol is shipped in one gallon polyethylene jugs, five




gallon DOT  17E steel pails and 55-gallon DOT 17E steel drums.  1,2-Pro-




panediol is shipped in one-gallon glass jugs, and 5- and 55-gallon




DOT  17E UCC #2 phenolic-lined steel pails and drums.  1,4-Butanediol is




available in 50, 100, and 400 pound lined steel drums and in  ten pound




packages (GAF, n.d.).






      2.   Personnel Exposure





      Respiratory protective equipment would not be needed unless workers




 are exposed to heated glycols.   When handling ethylene glycol,  Dow (1978a)




 suggests workers wear clean, body-covering protective clothing.  For




                                     213

-------
1,2-propanediol, no protective clothing is usually necessary (Dow,




1978b) .  For neither glycol is eye protection normally necessary -








     3.  Accident Procedures





     Union Carbide (.1976) suggests that small spills of ethylene glycol




should be flushed with large quantities of water.   Larger spills should




be collected for disposal.   To dispose, they suggest mixing the  waste




glycol with a flammable solvent and incinerating in a furnace  where  per-




mitted under Federal, State, and Local regulations.   Dow (1978a&b) sug-




gests flushing spills of ethylene glycol or 1,2-propanediol with water




and soaking up with absorbent material; large spills should be diked




and pumped into suitable containers.   As an alternate to incinerating,




Dow suggests recovering the spilled glycol with a  vacuum truck and re-




turning it to the plant for reprocessing.
                                    214

-------
                   V.  EXPOSURE AND EFFECTS POTENTIAL


     The most likely source of direct human contact with the glycols


is by ingestion.  1,2-Propanediol is a GRAS (generally recognized as


safe) food additive; the estimated daily intake by persons 2-65 years


old is 349 mg or 6 mg/kg (FASEB, 1973).  1,2-Propanediol is also used


in some oral drug preparations.  1,3-Butanediol is approved as a di-


rect food additive for flavoring substances; daily intake has not been


estimated.  For the other glycols, which are not approved as direct


food additives, ingestion occurs only during accident or suicidal at-


tempts.  Ingestion of ethylene glycol, usually as antifreeze, has ac-


counted for about 40-60 deaths per year (Haggerty, 1959).


     Dermal contact is another source of human exposure to the glycols.


1,2-Propanediol and 1,3-butanediol are used in some cosmetic formula-


tions, such as hand creams.  Occupationally, exposure to the skin is


possible during production and handling, but few reports of adverse


effects by this route were found in the literature.  Eczematous derma-


titis developed in a worker exposed to aqueous ethylene glycol solutions


during eyeglass manufacture (Dawson, 1976) .


     Inhalation is not a significant route of human exposure to the


glycols.  Their vapor pressures are low at ambient temperatures (Tables


2-4), so only at elevated temperatures will a potential inhalation


hazard exist.  Only one industrially-related report involving adverse

                      «
effects of inhalation was located in the literature.  This involved the


heating of ethylene glycol in an electrolytic condenser factory where


14 of 38 workers suffered loss of consciousness, nystagmus, and/or lympho-


cytosis (Troisi, 1950; see section III-A-1-b) .
                                    215

-------
     Major sources of environmental contamination by ethylene glycol


and 1,2-propanediol are from the disposal of spent antifreeze and from


the runoff of de-icing fluids.  Demand for antifreeze was about 195-215


million gallons in 1977 (Anon, 1977h;  Anon, 1977d) but estimates are


unavailable on the percentage of annual demand which eventually is dis-


posed of directly into the environment.  Limited monitoring data indicate


that entry of ethylene glycol and 1,2-propanediol into the environment


during production is a minor source (section II-D-1).  No environmental


monitoring data are available for the butanediol isomers.  The 2,3-


isomer occurs in the environment as an endproduct of fermentation by


several strains of enteric bacteria, such as Aerobacter, Klebsiella,


and Serratia (section II-D-5).  Another source of environmental exposure


by the glycols is during transportation accidents.


     The glycols present from any source are not likely to persist.  As


discussed in section II-F, they are subject to moderately rapid breakdown


by soil, water, and sewage microorganisms.  In the case of an accidental


spill into surface waters, a BOD problem could exist.  For example, Price


et al. (.1974) found that in freshwater, the bio-oxidation on day five


was 34% complete for ethylene glycol and 62% for 1,2-propanediol and on


day 20, 79% of the 1,2-propanediol had been biologically oxidized and


all of the ethylene glycol had disappeared.  Evans and David (1974)


reported complete degradation of ethylene glycol within three days in

                      c
river waters at 20°C.  It is possible that this relatively rapid rate


of degradation, imposing a high oxygen demand, might possibly kill


aquatic organisms through dissolved oxygen (DO) depletion.  Lower rates


of degradation were reported by Lamb and Jenkins  (1952) (on day five,
                                    216

-------
12.5% of theoretical oxygen demand was satisfied for ethylene  glycol




and 2.2% for 1,2-propanediol)  which would not likely result  in DO  de-




pletion.




     Fuller et al. (1976). evaluated the potential of several hundred




organic chemicals, including ethylene glycol  and 1,2-propanediol  to




enter the atmosphere and pose a toxicological threat.   They  used a scor-




ing system based on production, volatility,  and toxicity;  a  maximum




score of 125 was possible.  The higher the score, the more likely  a




toxicological threat.  A final score of 11 was assigned to ethylene gly-




col and 5 was assigned to 1,2-propanediol.  The scoring system is  most




useful when comparing chemicals.  Examples of other final  scores are




as follows:  acetonitrile, 23; acrotein, 21; aerylonitrile,  20; n-butyl




alcohol, butane,  7; ethylene, 9; and propylene, 4.
                                    217

-------
                             TECHNICAL SUMMARY

Production and Use


     The subject compounds contain two hydroxyl groups attached to car-

bon atoms in an aliphatic carbon chain.  The following glycols are dis-

cussed in this report:  ethylene glycol, 1,2-propanediol (propylene gly-

col) , 1,3-propanediol, 1,3-butanediol, 1,4-butanediol, 2,3-butanediol,

and 1,2-butanediol.  Of these, the most important commercially is ethylene

glycol.  About 3.7 billion pounds of this glycol were produced in the

U.S. during 1977 by the hydration of ethylene oxide or by the acetoxyla-

tion of ethylene.  It was produced at 16 sites by 12 manufacturers.

Ethylene glycol is used in antifreeze (45% of production). , polyester fi-

ber (35%) , alkyd and polyester resins (.4%) , latex paints, and other emul-

sions (.1%) as well as for export (.8%) and in a variety of miscellaneous

uses (7%) (MCP, 1977).

     Domestic production of 1,2-propanediol was about 0.5 billion pounds

during 1977.  This glycol is produced by five manufacturers at six sites

by the hydration of propylene oxide.  It has applications in polyester

resins (45%) , pet food (.12%) , exports (.12%) , food and Pharmaceuticals

(.11%), cellophane (7%) , tobacco (.7%), as well as other uses (6%) (Anon,

1977f) .  1,2-Propanediol is a generally recognized as safe (GRAS) food

additive.  1,3-Propanediol is of minor commercial importance, being re-

covered in commercial glycerol plants when there is sufficient demand
                      ?
(Budke and Banerjee, 1971) .

     1,4-Butanediol is manufactured in the U.S. by three producers at

four sites; total domestic demand is 175 million pounds.  It is used in

the production of tetrahydrofuran (.57%) , acetylenic chemicals (.26%) ,
                                     218

-------
polyurethanes (9%) , and polybutylene terephthalate (7%)  (Brownstein  and




List, 1977).  1,2- and 1,3-Butanediols are produced by  two  manufacturers




and 2,3-butanediol is produced by another manufacturer;  figures  for




plant capacity or total demand are unavailable.   The 1,2- and 1,3-isomers




are used primarily in the production of polymeric plasticizers while the




2,3-isomer is used as a solvent, a humectant and a coupling agent.








Environmental Exposure





     Only limited data are available on sources  of entry of the  glycols




into the environment.  Ethylene glycol and 1,2-propanediol  were  identified




as components of the wastewater from production  facilities  (Zeitoun  and




Mcllhenny, 1971); environmental entry from this  source  will depend on




the extent of waste stream treatment and on whether spent glycol re-




cycling is practiced.   The major source of environmental contamination




by ethylene glycol and 1,2-propanediol is likely from the disposal of




spent antifreeze, but no estimates are available which  assess the magni-




tude of this problem.  Runoff of de-icing fluids which  are  sprayed,  for




example, on runways and airplanes, is another source of environmental




contamination.  2,3-Butanediol occurs in the environment as an endproduct




of fermentation by several strains of enteric bacteria.




     The glycols are capable of being degraded by a variety of micro-




organisms.  Ethylene glycol (2 or 10 mg/5,) was biodegraded  completely in




three days when tested'in four types of river water at  20°C (Evans  and




David, 1974).  When added to adapted activated sludge,  most C97%) of a




sample of ethylene glycol was degraded over 120 hours (Fitter, 1976).




Price et al. (1974) found that the bio-oxidation of ethylene glycol  (up




to 10 mg/fc) was 34%, 86%, 92%, and 100% complete after 5, 10, 15, and 20
                                    219

-------
days, respectively, when settled domestic wastewater was used as seed;




using synthetic seawater as seed, bio-oxidation was 20,  60,  65, and 77%




complete after 5, 10, 15, and 20 days, respectively.  Price  et al.  (1974)




obtained similar results with 1,2-propanediol.   Bedard (1976) labeled




both ethylene glycol and 1,2-propanediol as readily degradable based on




BOD measurements using raw sewage seed.  For 1,4-butanediol, degradation




was 98.7% complete in 120 hours using adapted sludge (Fitter, 1976).




     The glycols are soluble in water in all proportions; when spilled




in a body of water they will sink then dissolve.  Since  their vapor pres-




sure is low, they will evaporate slowly.  Relatively rapid breakdown by




microorganisms will preclude environmental persistence.







Biological Effects of Ethylene Glycol





     Most cases of ethylene glycol intoxication in humans involved  acci-




dental ingestion.  Clinical signs may include nausea, hypertension,




tachycardia, cardiopulmonary failure, renal impairment,  and  coma; oxalate




crystals, formed by oxidation of ethylene glycol, are usually in the




urine.  The lethal dose in humans is about 1.4-1.6 g/kg  or about 100 ml




(Lavelle, 1977) .




     Ethylene glycol is oxidized to glycolaldehyde, which is further




oxidized to glycolate, then to oxalate and CC>2.  Glycolate has been




identified as the specific toxic agent in acute ethylene glycol poisoning




in the rat (Chou and Richardson, 1978) and the monkey (Clay  and Murphy,




1977) .




     In rats given i.v, doses of labeled ethylene glycol, 60% of the




dose had been eliminated within 24 hours as lltC02 in expired air and
                                    220

-------
  C-labeled compounds in the urine; the rest was widely distributed in




the tissues.  In monkeys, 15% was excreted in urine and expired air within




four hours.  Unchanged ethylene glycol and glycolic acid were the most




important urinary products (McChesney et al., 1971).




     The oral lethal dose averages 8.3-15.3 g/kg in mice, 6.1-8.5 g/kg




in rats and 6.6-8.1 g/kg in guinea pigs.  Comparable values were obtained




following parenteral administration.  Signs include weakness, loss of




muscular coordination, prostration, and coma (Laug et al.,  1939).  Mi-




croscopic findings include deposition of calcium oxalate crystals in the




kidney; ethylene glycol has been used as a model to study renal hyper-




oxaluria or to induce oxalate lithiasis.




     Renal changes are frequently noted with chronic ethylene glycol ad-




ministration.  Rats given 1% (but not 0.5%) ethylene glycol in the drink-




ing water for 120-130 days had oxalate crystals in the urine (Hanzlik




et al., 1931).  In three macaque monkeys given 37-152 g/kg ethylene gly-




col over 13 to 157 days, renal changes were proportional to the dose




(Roberts and Seibold, 1969); these changes included deposition of calcium




oxalate in the proximal tubule, necrotic epithelial cells,  and occasional




focal granulomas.  Few chronic effects other than renal changes have been




noted.  The addition of ethylene glycol to the diet of three rhesus mon-




keys at 0.2 or 0.5% for three years produced no observable toxic effects




(Blood et al., 1962).




     Several inhalation studies were carried out to evaluate the effects




of a possible leak of ethylene glycol from the heat exchanger system in




spacecraft.  Inhalation of 10 or 57 mg/m3 ethylene glycol for eight hours/




day, five days a week for 90 days was without effect in rats, guinea




pigs, rabbits, dogs, or monkeys; continuous exposure to 12 mg/m3 for 90







                                    221

-------
days resulted in eye irritation (Coon et al., 1970).  Chimpanzees contin-



uously exposed to 256 mg/m3 of ethylene glycol aerosol for 28 days showed



some impairment of auditory and visual discrimination compared to pre-



exposure levels.



     A single s.c. dose of 1 or 10 mg (~LDsQ) ethylene glycol elicited



no carcinogenic effect in mice after 15 months (WARF, 1970) .   In rats



receiving 30-1,000 mg/kg ethylene glycol s.c. twice a week for one year,



tumor incidence was comparable to that found in controls (Mason et al.,



1971) .  Skin painting of mice with ethylene glycol up to 86  times



 (Berenblum  and Haran,  1955) or twice  a week for life  (Deringer,  1962)



 did  not  result  in excess  tumor incidence compared  to controls.



     For aquatic organisms, the TL  (median tolerance limit)  or the LCsg
                                  m


was >20,000 mg/5, for brine shrimp, >100 mg/£ for brown shrimp, and



>18,500 mg/& for rainbow trout.






Biological Effects of Propylene Glycols




     1,2-Propanediol (propylene glycol) is classed as a Generally Recog-



nized as Safe (GRAS) food additive by the FDA.  Few reports of adverse



effects to humans have appeared in the literature; four case  studies



were located on suspected intoxication due to 1,2-propanediol ingestion.



Signs included arrhythmia, tachypnea, stupor, and lactic acidosis (Martin



and Finberg, 1970; Gate and McGlothlin, 1976).  Following an  oral dose



to three volunteers (1 g/kg), 20-25% of the dose was eliminated in the



urine after ten hours  (Hanzlik et al., 1939a) .  The potential for allergic



reaction to contact with this chemical has been noted by several in-



vestigators .
                                    222

-------
     In laboratory animals, 1,2-propanediol was rapidly absorbed  follow-




ing oral dosing or injection (Van Winkle,  194lb;  Lehman and Newman,




1937).  Up to 45% of the dose was excreted in the urine within 24 hours.




     1,2-Propanediol is metabolized through lactaldehyde,  methylglyoxal,




and then lactic acid or pyruvic acid (Ruddick,  1972) .   It  is further




oxidized through the tricarboxylic acid cycle or through the glycolytic




pathway, the latter contributing to glycogen formation. The glycogenic




effect of 1,2-propanediol has been noted by many investigators (Hanzlik




et al., 1939a and b; Giri et al., 197Q; Whittman and Bawin, 1974).




     The LDso for orally administered 1,2-propanediol  averages 14-25




g/kg in mice, 22-29 g/kg in rats and 18-20 g/kg in guinea  pigs.   This




glycol is more acutely toxic after injection than  by  oral administration.




Large doses of 1,2-propanediol resulted in loss of equilibrium,  central




nervous system depression, and respiratory failure (Laug et al.,  1939;




Giri et al., 1970; Seidenfeld and Hanzlik, 1932).  Following i.v. injec-




tion of 1,2-propanediol to rabbits, there was a dose dependent increase




in the number of circulating polymorphs and decrease in the number of




lymphocytes (Brittain and D'&rcy, 1962).  No hematological or other




changes were noted in rats fed diets containing 5% 1,2-propanediol for




15 weeks (Gaunt et al., 1972).  Other investigators failed to note con-




sistent chronic effects of 1,2-propanediol treatment.   No  pathological




changes were reported in rats given the glycol at levels up to 10% in




the drinking water for'up to 140 days (Weatherby and Haag, 1938; Seidenfeld




and Hanzlik, 1932).  Gaunt et al. (1972) reported that of  up to 5%




1,2-propanediol in the diet of Charles River rats for two  years was




without effect on mortality, body weight gain, hematology, urinary cell
                                     223

-------
excretion, renal chemistry, organ weights, or pathology.   In dogs re-




ceiving 8 or 20% 1,2-propanediol in the diet for two years,  Weil et al.




(1971) found no effects on the following parameters:  mortality, body-




weight change; diet utilization, or water consumption, micropathology;




liver, kidney, and spleen weights; and most values for blood,  urine,




and biochemical parameters.  Some erythrocyte destruction occurred in




dogs receiving the higher level of 1,2-propanediol.




     1,2-Propanediol was not teratogenic at daily oral doses up to 1,55Q~




1,600 mg/kg administered during days 6-15 of gestation to mice, rats, or




hamsters.  No teratogenic effect was noted in rabbits similarly treated




on days 6-10 of gestation with up to 1,23G mg/kg (FDKL, 1973a) .  No




carcinogenic potential has been attributed to 1,2-propanediol  administered




orally to mice (57, in the diet for two years) , by injection  to mice




(0.5-1.0 ml weekly for 88 weeks; single injection of 1 mg) or  by skin




painting to mice (10-100% 1,2-propanediol daily for life).




     No positive mutagenic effects were noted in a host-mediated assay




using mice receiving 30, 2,500, or 5,000 mg/kg 1,2-propanediol orally




followed by an i.p. dose of an indicator organism (Salmonella  typhimurium




or Saccharomyces cerevisiae) (Litton Bionetics,  1974).  Mice  receiving the




highest dose daily for five days showed a "weak or questionable positive"




result with one strain of Salmonella.  Cytogenetic assays of somatic




cells from rats given up to 5,000 mg/kg of 1,2-propanediol showed no




chromosomal aberrations.  Negative results were also obtained  in in vivo




tests with human embryonic lung cultures.  A dominant lethal gene test




with rats given 1 or 5 doses of up to 5,000 mg/kg 1,2-propanediol showed




no evidence of mutagenicity (Litton Bionetics, 1974).
                                    224

-------
     In rainbow trout, the LC5Q is between 50,000-100,000 mg/£ (Hann and



Jensen, 1974) and in brine shrimp, the 24 hour TL  is greater than 10,000
                                                 m


mg/i (Price et al., 1974).



     1,3-Propanediol has not been studied as extensively as the 1,2-



isomer.  The 50% fatal dose to rabbits was 4.2-7.4 g/kg, comparable to



1,2-propanediol.  By i.m. injection, the 50% fatal dose was 6.3-7.4 g/kg,



which is about twice that for 1,2-propanediol (Van Winkle, 1941).  Fischer



et al.  (1949) reported an LDso °f 6.4 g/kg for mice given 1,3-propanediol



orally.  1,3-Propanediol has no glycogenic action in rats, in contrast



to 1,2-propanediol.  In a 15 week study, rats given 5 or 12% 1,3-pro-



panediol in the diet or 5.3 or 10.6 g/kg daily by gavage showed a reduced



growth  rate, particularly those given the dietary diol.  Based on a



similar study by Van Winkle (1941), twice as much 1,2-propanediol is



required to produce the same effect as 1,3-propanediol.





Biological Effects of Butylene Glycols




     For 1,2-butanediol, few studies on biological effects are available.



The LD,.0 to rats is about 16 g/kg  (Rowe, 1963).   In large oral doses to



rats 1,2-butanediol caused narcosis, gastrointestinal irritation and per-



ipheral vasodilation.  Inhalation of an aerosol of 1,2-butanediol by rats



for 7 hours was without effect.  I.V. injection of up to 1 g/kg  to dogs



had no  effect.  It was not irritating to the skin of rabbits  (Dow Chemical



Co., in Rowe, 1963).  1,2-Butanediol showed a glycogenic effect  in starved



rats in contrast to the 2,3- and 1.4- isomers(0pitz, 1958).   Continuous



infusion of 0.1-0.3 g/kg/hr for more than 10 hours resulted  in diminished



muscle  tone in rabbits  (Strack et al., 1960).
                                     225

-------
     Little information is available on the biological effects of 2,3-




butanediol.  Fischer et al. (1949) reported that the oral LD   to mice




was 8.9 g/kg.  2,3-Butanediol was without narcotic effects in rats  (up




to 1.4 g/kg tested) and mice (0.5 g/kg tested)(Marcus et al., 1976; Menon




et al., 1973).




     1,3-Butanediol has been extensively studied as a synthetic metabolizable




source of energy.  Several studies in humans showed that isocaloric sub-




stitution of 1,3-butanediol for starch provided an adequate source of




dietary calories.  No toxic effects have been reported in a series of




nutritional and metabolic studies in humans.  Volunteers receiving the diol




had a decreased negative nitrogen balance and a decreased level of blood




glucose.  Up to 10% of the caloric intake of 1,3-butanediol appears to be




utilized (Kies et al., 1973; Tobin et al., 1975).




     Several studies in rats have confirmed that 1,3-butanediol is metabolized




via g-hydroxybutyric acid  (Gessner et al, 1960; Mehlman.et al., 1971 a & b;




Tobin et al., 1972; Rosmos et al., 1975).  In rats, 1,3-butanediol resulted




in an elevation of blood glucose in acute studies, but a lowering in chronic




studies.




     The oral LD   values for 1,3-butanediol average 23 g/kg in mice, 23-




30 g/kg in rats and 11.5 g/kg in guinea pigs.  By subcutaneous injection, the




LD^Q is 16.5 g/kg in mice and 20 g/kg in rats.  Exposure for 8 hours to




saturated vapors was without effect in rats (Smyth et al., 1951).  Large




parenteral doses of 1,3-butanediol have a narcotic effect; it appears to act




as a muscle relaxant rather than as a depressant of brain activity  (Sprince




et al., 1966).  Bornmann (1954 a & b; 1955) reported marked diuresis in rats




given an acute dose of 10.1 g/kg orally or up to 20% in the drinking water




twice a week for 96 days.  Several investigators have reported a decrease in




body weight gain when levels of 1,3-butanediol were (in most cases) 20% or
                                  226

-------
higher in the diet.  However, no histological or other changes were attributed




to butanediol treatment.  Administration of 10.6 mg/kg every 3-4 days to




rats for 45-185 days was without effect on blood count or organ histology




(Kopf et al., 1950).  No adverse effects on weight gain, survival, hematology,




urinalyses or histology were noted in rats fed diets containing 1, 3 or 10%




1,3-butanediol for 2 years or in dogs fed diets containing 0.5, 1 or 3% for




2 years (Scala and Paynter, 1967).  In a multigeneration feeding study, rats




received 5, 10 or 24% 1,3-butanediol in a semi-synthetic diet  (FDRL, 1973b).




The growth rate of males was slightly depressed in all generations.  There




was a gradual decrease in the pregnancy rate in 5 successive mating cycles in




F.. females which was attributed in part to the added stress of the semi-




synthetic diet.  No gross or pathological changes noted at autopsy of F




rats were attributed to treatment; no teratogenic effects were noted in litters




of F~ rats.  No teratogenic effects were noted in litters from rats and dogs




receiving up to 20-24% 1,3-butanediol prior to and during pregnancy or in




litters from rabbits receiving 1.3-5.4 g/kg daily on days 6-18 of gestation




(FDRL, 1973b).  A dominant lethal test and cytogenetic test revealed no




adverse effects on the genetic material in rats (FDRL, 1973b).




     1,4-Butanediol is more acutely toxic than the other butanediol isomers.




Its oral LDj.n value ranges from 1.2-2.5 g/kg in laboratory animals.  1,4-




Butanediol is a central nervous system depressant; this effect is mediated




through its metabolite, gamma-hydroxybutyrate.  Signs of intoxication




include narcosis, bradycardia, analgesia and akinesia  (Hinricks et al., 1948;




Zabik et al., 1974; Menon et al., 1973).






Regulations and Standards





     The Food and Drug Administration regulates'1,3-butanediol and 1,2-




propanediol as both direct and  indirect  food additives  and  ethylene glycol,
                                  227

-------
1,4-butanediol and butylene glycol (isomer unspecified)  as indirect food




additives.  The FDA has proposed exposure limits for ethylene glycol,  ethylene




oxide and ethylene chlorohydrin in certain drug products and medical devices.




There are no specific EPA regulations governing water or air quality with




respect to the title glycols.  Propylene glycol can be used as an inert




solvent or cosolvent in pesticide formulations.  None of the glycols




considered in this report are specifically regulated by  the Occupational




Safety and Health Administration or by the Department of Transportation.
                                     228

-------
                                BIBLIOGRAPHY
ACGIH, (1977) "Threshold Limit Values for Chemical Substances in Workroom
     Air Adopted by the ACGIH for 1977," Cincinnati, Ohio.

Aldrich, (1978), personal communication to Aldrich Chemical Co., Inc.,
     Metuchen, N. J., price quotation.

Allen, R. C., Meier, H., and Hoag, W. C., (1962)  "Ethylene Glycol Produced
     by Ethylene Oxide Sterilization and its Effect on Blood-Clotting
     Factors in an Inbred Strain of Mice,"  Nature, 193(4813):387-388.

Anon., (1930) "Possible Death from Drinking Ethylene Glycol (Prestone),"
     J. Am. Med. Assoc., 94:1940.

Anon., (1976a) "DuPont Will Expand Butanediol and THF,"   Chem. Mark. Rep.,
     209(5):3,44.

Anon., (1976b) "Chemical Profile:  Ethylene Glycol," Chem. Mark. Rep.,
     January 1, 1976:9.

Anon., (1977a) "Ethylene Oxide and Glycol-Shell Development Co.," Hydroc.
     Proc., November 1977:159.

Anon., (1977b) "Ethylene Oxide and Glycols-Snamprogetti," Hydroc. Proc.,
     November 1977: 161.

Anon., (1977c) "Aliphatic Organics:  Ethylene Glycol,"  Chem. Mark. Rep.,
     Oct. 17, 1977:31.

Anon., (1977d) "Key Chemicals:  Ethylene Glycol," Chem. Eng.  News, Dec. 5,
     1977:12.

Anon., (1977e) "New Ethylene Oxide Plant in Operation at  Louisiana Site,"
     Oil Gas J., June 13, 1977:23.

Anon., (1977f) "Chemical Profile:  Propylene Glycol,"  Chem.  Mark. Rep.,
     211(18):9.

Anon., (1977g) "CPI News Briefs,"  Chem. Eng., April 25,  1977:148.

Anon., (1977h) "Detergent Olefins Unit Starts For Shell at Geismar Complex,"
     Chem. Mark. Rep., October 24, 1977:4,17.

Anon., (1977i) "Butanediol Plant Brought on Stream,"  Rubber  World,
     February 1977:10.

Anon., (1978a) "ICI Plans U. S. Ethylene Oxide Plant,"  Chem. Eng. News,
     May 1, 1978:10.
                                    229

-------
Anon.,  (1978b) "51 Canada,"  International  Environment  Reporter,  Bureau of
     National Affairs,  Inc., Washington, D.  C.

Bachem,  C.,  (1917)  ["Investigations  on  Glycols  and their' Use in  Pharmacy
     and Medicine," ] Med .  Klin. ,1:7  (German), .

Bachmann,  E. and  Golberg,  L.,  (1971)  "Reappraisal of  the Toxicology of
     Ethylene Glycol:III.  Mitochondrial Effects,"  Fd.  Cosmet.  Toxicol.,
     9:39-55.

Bahadur, K.  and Dube,  J.  N.,  (1959)  "Production of 2,3-Butanediol by
     Serratia marcescens  from  Agricultural Waste Materials," Indian J.
     Appl.  Chem.,  22:99-102.

Bahadur, K. and Ranganayaki, S.,  (1961) "Role of  Intracellular and^Extra-
     cellular Enzymes in the Production of Acetoin and 2,3-Butanediol
     by  Bacillus polymyxa," Japan. J. Microbiol., 5:11-15.

Baldwin, R. W.,  Cunningham, G. J., Smith, W. R. D., and  Surtees,  s.  J.,
     (1968) "Carcinogenic Action of 4-Acetamidostilbene  and  the N-Hydroxy
     Derivative in the Rat,"  Brit. J. Cancer, 22:133-144.

Bartsch, W., Sponer, G,, Dietmann, K., and Fuchs, G.,  (1976) "Acute
     Toxicity of Various Solvents in  the Mouse and Rat," Arzneim. Forsch.,
     26(8):1581-1583.

Bedard,  R.  G., (1976) "Biodegradability of Organic Compounds," NTIS  PB-264
     707, 84 pp.

Belue,  G.,  (1974) "High-Pressure Liquid Chromatography of Polyhydric
     Alcohols on Silica Gel Columns,"  J. Chromatogr., 100:233-235.

Berenblum,  I. and Haran, N., (1955) "The Initiating Action of Ethyl
     Carbamate (Urethane) on Mouse Skin,"  British J. Cancer, 9:453-456.

Berman,  L.  H., Schreiner, G. E., and  Feys,  J., (1957) "The Nephrotoxic
     Lesion of Ethylene Glycol," Ann. Internal Med., 46:611-619.

Blood,  F. R., Elliott, G. A., and Wright, M. S.,  (1962)  "Chronic  Toxicity of
     Ethylene Glycol in the Monkey,"  Toxicol. Appl. Pharmacol.,  4:489-491.

Bonnardeaux, J.  L., (1971) "A Comparison of  the Effects  of Three  Organic
     Solvents:  Dimethyl Sulfoxide, Formamide, and Propylene Glycol, on
     Spontaneous Activity of Isolated Smooth Muscle,"  Can.  J. Physiol.
     Pharmacol., 49 (7):632-641.

Bonner,  J.  M. , Hess, G. S., Otchere,  E. 0.,  and Young, J. W. ,  (1974)
     "Physiological Effects of 1,3-Butanediol Fed to Cattle,"  J. Dairy  Sci
     58(l):56-62.

Bornmann, G., (1954a) ["Physiological Properties  of Glycols  and Their
     Toxicity.   Part 1,"]  Arzneimittel-Forsch.,  4:643-646 (German).

Bornmann, G., (1954b) ["Physiological Properties  of Glycols  and Their
     Toxicity.   Part 2,"]  Arzneimittel-Forsch.,  4:710-715 (German).


                                    230

-------
Bornmann, G.,  (1955)  ["Physiological Properties of Glycols and Their
     Toxicity.  Part  3,"] Arzneimittel-Forsch., 5"38  (German).

Braun, H. A. and Cortland, G. F., (1936) "The Toxicity of Propylene Glycol,"
     Am. Pharm. Assoc., 25:746-749.

Brittain, R. T. and D'Arcy, P. F.,  (1962) "Hematologic Effects Following
     Intravenous Injection of Propylene Glycol in the Rabbit,"  Toxicol.
     Appl. Pharmacol., 4:738-744.

Brock, T. D.,  (1970) Biology of Microorganisms, Prentice-Hall, Inc.,
     Englewood Cliffs, N.J.

Brown, D. J. and Conine, D. L., (1971) "Comparative Toxicity of the Optical
     Isomers of Propylene Glycol,"  Pharmacologist, 13(2):240.

Brownstein, A. M.,  (1974) "Energy Crisis Impacts on Ethylene Glycol Trends,"
     Hydroc. Proc., June 1974:130-133.

Brownstein, A. M. and List, H. L.,  (1977) "Which Route to 1,4-Butanediol,"
     Hydroc. Proc., Sept. 1977:159-162.

Budke, C. C. and Banerjee, D. K., (1971) "Glycols and Polyhydric Alcohols,"
     in Encycl. Indus. Chem. Analysis, 12:533-607.

Cantoni, C., Molnar, M. R., and Renon, P.,  ["Studies  on  the Production of
     Acetoin,  Diacetyl, and 2,3-Butylene Glycol by Micrococci and Lacto-
     bacilli,"] Arch. Vet. Ital., 16(2):85-93  (Italian).

Gate, J. C. and McGlothlin, (1976)  "Propylene Glycol  Overdosage Associated
     With Lactic Acidosis," Clin. Chem., 22(7):1199.

Celanese Chemical Co., (1977) "Product Index — Intermediates," Chem. Wk.,
     October 26, 1977:63.

Celanese Chemical Co., (1978) personal communication, price quotation.

Champion, M. H., (1970) "Collaborative Study on the GLC  Determination of
     Propylene Glycol in Cosmetics," J. Assoc. Offic. Anal. Chem., 53:82-84.

Child, J. and Willetts, A., (1978)  "Microbial Metabolism of Aliphatic Glycols;
     Bacterial Metabolism of Ethylene Glycol,"  Biochim.  Biophys. Acta. ,
     538:316-327.

Chino, N., Awad, E. A., and Kottke, F. J.,  (1974) "Pathology  of Propylene
     Glycol Administered by Perineural and  Intramuscular Injection in Rats,"
     Arch. Phys. Med. Rehabil., 55:33-38.

Chou, J. Y. and Richardson, K. E.,  (1978) "The Effect of Pyrazole on
     Ethylene  Glycol Toxicity and Metabolism in the Rat," Toxicol. Appl.
     Pharmacol., 43:33-44.

Clay, K. L. and Murphy, R. C.,  (1977) "On the Metabolic  Acidosis of
     Ethylene  Glycol  Intoxication," Toxicol. Appl. Pharmacol.,  39:39-49.
                                     231

-------
Clegg, D. J.,  (1964) "The Hen Egg in Toxicity and Teratogenicity  Studies,"
     Fd. Cosmet. Toxicol., 2:717-727.

Coon, R. A., Jones, R. A., Jenkins, L. J., and Siegel, J.,  (1970)  "Animal
     Inhalation Studies on Ammonia, Ethylene Glycol,  Formaldehyde,
     Dimethylamine, andEthanol,"  Toxicol. Appl. Pharmacol.,  16:646-655.

Cosby, J. N.,  (1957) "Synthesis of Carbonyl Compounds,"  U.  S.  Patent
     2,808,429, October 1, 1957, 6 pp.

Cox, J. W. and Wilkes, M. C., (1975) "Reclamation of  Spent Glycol  by
     Distillation  in the Presence of a Catalytic Amount of Alkali  Metal
     Hydroxide," U. S. Patent 3,878,055, April 15, 1975, 11  pp.

Curme, G. 0. and Johnston, F.,  (1952) Glycols, American Chemical  Society
     Monograph Series, Reinhold Publishing Corp., N.Y.

D'Amato, F.,  (1948) "The Effect of Colchicine and Ethylene Glycol  on
     Sticky Chromosomes in Allium cepa," Hereditas, 34:83-103.

Davenport, R. F. and Griffith, M., (1969) "Effect of  Varying Levels of
     Dietary 1,3-Butanediol on Growing Chicks," Poultry Sci.,  48(4):
     1369-1371.

Davis, K. J. and Jenner, P. M., (1959) "Toxicity of Three Drug Solvents,"
     Toxicol. Appl. Pharmacol., 1:576-578.

Dawson, T. A., (1976) "Ethylene Glycol Sensitivity,"  Contact Dermatitis,
     2(4):233.

Dean, M. E. and Stock, B. H., (1974) "Propylene Glycol as a  Drug  Solvent
     in the Study  of Hepatic Microsomal Enzyme Metabolism in the Rat,"
     Toxicol. Appl. Pharmacol., 28(1):44-52.

Delphia, J. M. and Frierdich, M., (1973) "The Dose-Dependent Glycogen-
     Depleting Effect of 1,2-Propanediol on the Myocardium of  the Eight
     and One-Half  Day Chick Embryo," Res. Commun. Chem. Pathol. Pharmacol.,
     6(1):337-340.

Dewhurst, F. , Kitchen, D. A., and Calcutti, G., (1972) "The  Carcinogenicity
     of Some 6-Substituted Benzo(a)Pyrene Derivatives in Mice," British  J.
     Cancer, 26:506-508.

Deringer, M. K., (1962),"Response of Strain HR/De Mice to Painting with
     Urethan," J.  Nat. Cancer Inst., 29(6):1107-194.

Dow  Chemical Co.,  (1975) Sales  Specifications:  U.S.P- and Industrial
     Propylene Glycol, Midland, Michigan.

Dow  Chemical U.S.A., (1978a) Material Safety Data Sheet:  Ethylene Glycol
     (Regular), Midland, Michigan.

Dow Chemical U.S.A.,. (1978b)  Material Safety Data Sheets:   Propylene Glycol
     U.S.P.  and Propylene Glycol Industrial,  Midland,  Michigan.
                                    232

-------
Dow Chemical Co.,  (1978c) personal  communication,  price quotation.

Dubeikovskaya, L.  S., Asanova,  T. P.,  Rozina,  G.  Y., Budanova,  L. F.,
     Zenkevich, E. S., Revnova, N.  V.,  and Corn,  L. E., (1973)  ["Hygienic
     Significance  of Ethylene Glycol  in the Manufacture of  Some  Radio
     Components"]  Gig.  Truda i. Prof. Zabol., 10:1-4.

Dymsza, H. A., (1968) "The  Biological  Status and  Potential  of 1,3-Butylene
     Glycol," unpublished report prepared  for  Celanese Corp., 40 pp.

Dymsza, H. A., (1975) "Nutritional  Application and Implication  of
     1,3-Butanediol," Fed.  Proc., 34(12):2167-2170.

Dymsza, H. A. and  Adams, P.  K.,  (1969)  "Influence  of 1,3-Butanediol and
     Fatty Acid Esters on Rat Reproduction and Development," Fed. Proc.,
     28:363.

Efstathiou, C. H.  and Hadjiioannou, T.  P.,  (1975)  "Automatic Reaction Rate
     Method for the Determination of Vicinal Glycols with a Perchlorate Ion
     Selective Electrode,"  Anal. Chem.,  75(6):864-869.

Eichbaum, F. W. and Yasaka,  W.  J.,  (1976)  "Antiarrhythmic Effect of Solvents:
     Propylene Glycol, Benzyl Alcohol," Basic  Res. Cardiol., 71(4):355-370.

EPA,  (1974) Development  Document for  Effluent  Limitations Guidelines and
     New  Source Performance Standards  for  the  Major Organic Products
     Segment of the Organic Chemicals  Manufacturing Point Source Category,
     pp.  183-186,  EPA 440/1-74-009.

EPA, (1975) Development  Document for  Interim Final Effluent Limitations
     Guidelines and New  Source  Performance  Standards for the Significant
     Organic Products Segment of the Organic Chemical Manufacturing Point
     Source Category, pp. 117-119,  EPA 440/1-75/036.

Evans, W. H. and David,  E.  J.,  (1974)  "Biodegradation of Mono-, Di-, and
     Triethylene Glycols in River Waters Under Controlled Laboratory
     Conditions,"  Water  Res., 8(2):97-100.

Evans, W. H. and Dennis, A.,  (1973) "Spectrophotometric Determination of
     Low  Levels of Mono-, Di-,  and  Triethylene Glycols in Surface Waters,"
     Analyst, 98:782-791.

Farr,  G.  G. and Norris,  W.  E.,  (1971)  "Effects of  Various Osmotic Agents  on
      Elongation of Avena Coleoptile Segments," Texas. J.  Sci.,  23(2):253-264.

FASEB, (1973) "Evaluation of the Health Aspects of Propylene Glycol and
      Propylene Glycol Monosterate as  Food  Ingredients," Federation  of
      American Societies  for Experimental Biology,  Prepared  for  the  Food and
      Drug Administration, NTIS  PB-265  504, 16  pp.

Felts, M. F.,  (1969) "Effects of Exposure  to Ethylene Glycol  on Chimpanzees,"
      Proc.  5th Annual Conf.  on  Atm. Contain. Confined  Spaces,  16-18  September
      1969,  AMRL-TR-69-130-.105-124.
                                     233

-------
Fields, M. L. and Richmond, B., (1967) "Substrate Effect on 2,3-Butylene
     Glycol Production by Rhizopus nigricans and Penicillium expansum,"
     Appl. Microbiol., 15(6):1313-15.

Fisher, L., Kopf, R., Loeser, A., and Meyer, G., (1949) ["Chemical
     Structure and Pharmacological Effect of Glycols, Especially 1,3-
     Butylene Glycol,"] Zeit. Ges. Exptl. Med., 115:22-39 (German).

Flury, F. and Worth, W., (1934) ["The Toxicology of Solvents"] Arch.
     Gewerbepathol. Gewerbehyg., 5:52 (German).

Fonck-Cussac, Y., Aublet-Cuvelier, J. L., Fonck, J., and Delage, J., (1971)
      ["Ultrastructural Study of the Kidney in  Experimental Oxalosis
     Induced With Ethylene Glycol,"] Ann. Anat. Pathol., (Paris),
     16(2):153-158  (French).

Food and Drug Research Labs.,  Inc., (1973a) "Teratologic Evaluation of
     FDA 71-561 Propylene Glycol," prepared for the Food and Drug
     Administration, NTIS PB-223 822, 56 pp.

Food and Drug Research Laboratories, Inc., (1973b) "Reproductive and
     Genetic Studies with 1,3-Butylene Glycol," submitted to Celanese
     Chemical Co., N.Y.

Freifeld, M. and Hort, E. V.,  (1966) "1,4-Butylene Glycol and y-Butyrolactone,"
     Kirk-Othmer Encycl. Chem. Technol., (2nd  Ed.), 10:667-676.

Friedman, E. A., Greenberg, J. B., Merrill, J. P., and Dammin, G. J.,  (1962)
      "Consequences of Ethylene Glycol Poisoning," Am. J. Med., 32:891-902.

Fujino, H., Chino,  T., and Tamaki, I., (1965)  "Experimental Production of
     Labial and Lingual Carcinoma by Local Application of 4-Nitroquinoline
     N-Oxide," J. Nat. Cancer  Inst., 35:907-918.

Fuller, E. W., Jr., (1969) "Ethylene Glycol Toxicity:  A Review,"
     Medico-Legal Bull. No. 198, 18(10):l-8.

Fuller, B., Hushon, J., Kornreich, M., Quellette, R., Thomas, L., and
     Walker, P.,  (1976) "Preliminary Scoring of Selected Organic Air
     Pollutants," EPA-450/3-77-008a.

GAF,  (n.d.) 1,4-Butanediol, Dyestuff and Chemical Division, N.Y., Technical
     Bulletin 7543-077,,13 pp.

GAF,  (1977) personal  communication, price quotation.

Gardner, S., (1978) "Ethylene  Oxide, Ethylene  Chlorohydrin and Ethylene
     Glycol:  Proposed Maximum Residue Limits  and Maximum Levels of Exposure,"
     Fed. Reg., 43(122):27474-27483.
                                     234

-------
Gaultier, M., Conso, F., Rudler, M.,  Leclerc, J. p., and Mellerio,  F.,  (1976)
     ["Ethylene Glycol  Acute Poisoning,"] Eur. J. Toxicol. Environ. Hyg.,
     9(6):373-379  (French).

Gaunt,  I. F., Carpanini, F. M.  B., Grasso, P., and Lansdown, A. B.  G.,  (1972)
     "Long-Term Toxicity of Propylene Glycol in Rats," Fd. Cosmet.  Toxicol.,
     10:151-162.

Gebhardt, D. 0. E.,  (1968) "The Teratogenic Action of Propylene Glycol
     (Propanediol-1,2)  and Propanediol-1,3 in Chick Embryo," Teratol.,
     1:153-162.

Gershoff, S. N. and  Andrus, S.  B. ,  (1962) "Effect of Vitamin Bg and Magnesium
     on Renal Deposition of Calcium  Oxalate Induced by Ethylene Glycol
     Administration," Proc. Soc.  Exp.  Biol. Med., 109:99-102.

Gessner, P- K., Parke,  D. V.,  and Williams, R. T., (1960) "Studies  in
     Detoxication:80.   The Metabolism of Glycols," Biochem. J., 74:1-5.

Gessner, P. K., Parke,  D. V.,  and Williams, R. T., (1961) "Studies  in
     Detoxication:86.   The Metabolism of  1 ^-Labelled Ethylene Glycol,"
     Biochem. J.,  79(3):482-489.

Giesecke, D., Dirksen,  G., Guenzel,  R., and Baumer, G., (1975)  ["Glucogenic
     Effect of 1,2-Propanediol in Healthy Cows and in Phlorizinized Sheep"]
     Dtsch. Tieraerztl. Wochenschr.,  82(3):105-109 (German).

Giri, S. N., Peoples, S. A., and  Mull,  R. L.,  (1970) "Effects of 1,2-
     Propanediol on  Blood Glucose and Liver Glycogen of Young Rats,"
     Proc.  Soc. Exp. Biol. Med.,  133(2):593-596.

Gonzalez, C. F., Taber, W. A.,  and Zeitoun, M. A., (1972) "Biodegradation  of
     Ethylene Glycol by a  Salt-Requiring Bacterium," Appl. Microbiol.,
     24(6):911-919.

Gorban,  N.  S. and  Petrenko, M.  B.,  (1972)  ["Participation of Microorganisms
     in  the Transformation of  Propylene Glycol, a Component of Waste Waters,"]
     Mikrobiol. Zh., 34(5):571-575  (Ukranian).

Grabinska-Loniewska, A.,  (1974a)  "Studies on  the Activated Sludge  Bacteria
     Participating in  the  Biodegradation  of Methanol, Formaldehyde and
     Ethylene Glycol:   Part 1.   Isolation and  Identification," Acta Microbiol.
     Pol. Ser. B,  6(23):75-81.
                        ?
Grabinska-Loniewska, A.,  (1974b)  "Studies on  the Activated Sludge  Bacteria
     Participating in  the  Biodegradation  of Methanol, Formaldehyde and
     Ethylene Glycol:   Part II.  Utilization  of Various Carbon and Nitrogen
     Compounds," Acta.  Microbiol. Pol.  Ser.  B., 6(23):83-88.

Granich, M. and  Timiras, P. s., (1969)  "Toxicity of Propylene Glycol  Injected
      Into  the Air  Chamber  of  Embryonated  Chicken  Eggs:  Acute LD5Q, and
      Impaired  Hatchability of  Surviving Embryos," Fed. Proc.,  28:2715.
                                     233

-------
Griffin, W. C. and Lynch, M. J.,  (1972) "Polyhydric Alcohols,"  in  Furia,
     T. E., Handbook of Food Additives, CRC Press, Cleveland, Ohio.

Haggerty, R. J., (1959) "Toxic Hazards —Deaths from Permanent  Antifreeze
     Ingestion," N. E. J. Med., 261:1296 -1297.

Raines, J. R. and Alexander, M.,  (1975) "Microbial Degradation  of  Polyethylene
     Glycols," Appl. Microbiol., 29(5):621-625.

Hammarsten, G., (1956) "On Calcium Oxalate Stones," in Butt, A. J.  (ed.),
     Etiologic Factors in Renal Lithiasis, Charles C. Thomas, Springfield,
     Illinois.

Hann, R. W. and Jensen, P. A., (1974)  "Water Quality Characteristics of
     Hazardous Materials," Environmental Engineering Division,  Texas A & M
     Univ.

Hanzlik, P. J., Newman, H. W., Van Winkle, W., Lehman, A. J., and  Kennedy,
     N. K., (1939a) "Toxicity, Fate and Excretion of Propylene  Glycol and
     Some Other Glycols," J. Pharmacol. Exp. Therap., 67(1):101-113.

Hanzlik, P. J., Lehman, A. J., Van Winkle, W., and Kennedy, N.  K.,  (1939b)
     "General Metabolic and Glycogenic Actions of Propylene Glycol  and Some
     Other Glycols," J. Pharmacol. Exp. Therap., 67(1):114-126.

Hanzlik, P. J., Seidenfeld, M. A., and Johnson, C. C., (1931) "General
     Properties, Irritant and Toxic Actions of Ethylene Glycol," J. Pharmacol.
     Exptl. Therap., 41:387-406.

Harada, T. and Hirabayashi, T., (1968) "Utilization of Alcohols by Hansenula
     miso," Agric. Biol. Chem., 32(9):1175-1180.

Harper, C. A., (1975) "Fundamentals of Plastics and Elastomers" _in  C. A.
     Harper (ed.), Handbook of Plastics and Elastomers,  McGraw-Hill Book
     Co., N.Y., pp.1-1 to 1-121.

Harris, E. S., (1969) "Inhalation Toxicity of Ethylene Glycol," U.  S. Air
     Force Tech. Doc. Rep., 69(130):99-104.

Hatch, L. F. and Matar, S., (1977) "From Hydrocarbons to Petrochemicals.
     Part 6 —Petrochemicals from Methane," Hydroc. Proc., October  1977:153-163,

Hatch, L. F. and Matar, S., (1978a) "From Hydrocarbons to Petrochemicals:
     Part 10 — Chemicals from Ethylene," Hydroc. Proc.,  April 1978:155-166.

Hatch, L. F. and Matar, S., (1978b) "From Hydrocarbons to Petrochemicals:
     Part 12 — Chemicals from C^'s," Hydroc. Proc., August 1978:153-165.

Hinrichs, von A.,  Kopf, R., and Loeser, A., (1948) ["The Toxicology of
     1,4-Butanediol"] Pharmazie, 3:110-112 (German).

Holman, N. W.  and Mundy, R. L., (1976) "Analysis of Ethylene Glycol and
     Glycolaldehyde in Mouse Plasma,"  Fed. Proc., 35(3):534.


                                    236

-------
Isgrig, F. A. and Ayres, J. J. B.,  (1968) "Some Behavioral Effects  of Two
     Experimental Synthetic Nutrients," Psychopharmacol., 12(3):227-235.

Jank, B. E., Guo, H. M., and Cairns, V. W.,  (1974) "Activated Sludge Treatment
     of Airport Wastewater Containing  De-icing Fluids," Waier Res.,
     8:875-880.

Johnson,  E.L.,  (1978)  "Notice  of  Rebuttable Presumption Against  Registration
     and  Continued  Registration of Pesticide Products  Containing Ethylene
     Oxide," Fed. Regist.,  43(19):3801-3815.

 Jones, N. and Watson,  K.,  (1976)  "Ethylene Glycol and  Polyethylene Glycol
      Catabolism by a  Sewage Bacterium," Biochem. Soc.  Trans., 4(5):891-892.

 Kallgren, R. W., (1963) "Antifreezes and De-icing Fluids," Kirk-Othmer
      Encycl. Chem. Technol.,  (2nd Ed.), 2:540-561.

 Karel, L.,  Landing, R. H., and Harvey, T.  S.,  (1947) "The Intraperitoneal
      Toxicity of Some Glycols, Glycol Ethers, Glycol Esters, and Phthalates in
      Mice," J. Pharmacol. Exp. Therap., 91:338-347.

 Kawami, Y.  and Torikai, E., (1976)  ["Studies on Electrolytic Treatment of
      Waste  Water.  IV.  Electrolytic Treatment of Effluent Containing Ethylene
      Glycol,"] Osaka  Kogyo Gijutsu Shikensho Kiho,  27(4):249-255  (Japanese).

 Keith, W.C.,  Burk, E.H., and  Keith, C.D.,  (1961) "Olefin Oxidation With a  Solid
       Calcined  Catalyst," U.S. Patent  2,974,161, March  7, 1961,  4  pp.

 Kersting, E. J. and Nielsen,  S.  W.,  (1966) "Experimental Ethylene  Glycol
      Poisoning in  the Dog," Am.  J.  Vet. Res.,  27(117):574-582.

 Kies,  C., Tobin, R. B., Fox,  H.  M.,*  and Mehlman, M. A., (1973)  "Utilization
      of  1,3-Butanediol in Human  Adults," J. Nutr.,  103(2):1155-1163.

 Kirschbaum, B. B.  and Bosmann, H. B.,  (1974) "Acid  Hydrolase and  Glycoprotein:
      Glycosyl Transferase Activities  in Experimental Renal Disease in  the
      Rat,"  Chem.-Bio1. Interact., 8:207-216.

 Knyshova, S. P., (1968)  ["Specific Features of Biologic Action and the
      Hygienic  Significance of 1,4-Butyndiol and 1,4-Butanediol"]  Gig.  Sanit.,
      33(1):37-41 (Russian).

 Konradova,  V., Vavrova, V., and Janota, J., (1978)  "Effect  of  the Inhalation
      of  a Surface  Tension-Reducing Substance (Propylene Glycol) on the
      Ultrastrueture of the Epithelium of the Respiratory Passages in Rabbits,"
      Folia  Morphol.  (Praha),  26(l):28-34.

 Kopf,  R.,  Loeser,  A., and Meyer, G.,  (1950) ["Investigation on the Pharmacology
       and Toxicology of a Polyvalent Alcohol (1,3-Butylene  Glycoll"],  Arch.
       Exper.  Path.  Pharmacol., 210:346-360  (German).

 Kornberg, H.  L. and Gotto, A. M., (1959) "Biosynthesis of  Cell Constituents
       from C2-Compounds," Nature, 183(4678):1791-1792.


                                     237

-------
Lamb, C. B. and Jenkins, G. F., (1952) "B.O.D. of Synthetic Organic  Chemicals,"
     Proc. 7th Ind. Waste Conf. Perdue Univ., 79:326-339.

Lavelle, K. J., (1977) "Ethylene Glycol Poisoning," J. Ind. State Med. Assoc.,
     70(5):249-252.

Levy, R. I., (1960) "Renal Failure Secondary to Ethylene Glycol  Intoxication,"
     J. Am. Med. Assoc., 173:1210-1213.

Latven, A. R. and Molitor, H., (1939) "Comparison of the Toxic,  Hypnotic
     and Irritating Properties of Eight Organic Solvents," J. Pharmacol.  Exp.
     Therap., 65:89-103.

Laug, E. P., Calvery, H. 0., Morris, H. J., and Woodard, G.,  (1939)  "The
     Toxicology of Some Glycols and Derivatives," J. Indust. Hyg. Toxicol.,
     21:173-201.

Lehman, A. J. and Newman, H. W., (1937) "Propylene Glycol:  Rate of  Metabolism,
     Absorption, and  Excretion, with a Method For Estimation  in  Body Fluids,"
     J. Pharm. and Exper. Therap., 60:312-322.

Liao, L. L. and Richardson, K. E., (1972) "The Metabolism of  Oxalate Precursors
     in Isolated Perfused Rat Livers," Arch. Biochem. Biophys.,  153:438-448.

Little, A. D., Inc.,  (1974) "A Modal Economic and Safety Analysis of  the
     Transportation of Hazardous Substances in Bulk," prepared for U. S.  Dept.
     Commerce, Maritime Adm., NTIS COM-74-11271/5WP, 264 pp.

Litton Bionetics, Inc.,  (1974) "Mutagenic Evaluation of Compound FDA 71-56:
     Propylene Glycol," prepared for the Food and Drug Administration,
     NTIS  PB-425 450, 88 pp.

Lowenheim, F. A. and  Moran, M. K., (1975) Faith, Keyes, and Clark's  Industrial
     Chemicals, 4th  Ed., Wiley-Interscience, N. Y.

Lyon, E.  S., Borden,  T. A., and Vermeulen, C. W.,  (1966) "Experimental
     Oxalate Lithiasis Produced With Ethylene Glycol," Invest. Urol.,
     4(2):143-151.

MacCannell, K.,  (1969) "Hemodynamic Responses to Glycols and  to  Hemolysis,"
     Can.  J. Physio1. Pharmacol., 47:563-569.

 Mackenzie, J.B.,  and Mackenzie, C.G.,  and Reiss, 0.,  (1968)  "Regulation  of
      Cell Lipid  Metabolism and Accumulation. VII.  Increase by Glycerol of
      the Polar Lipid and Triglyceride  Content of Cultured Cells, Proc.
      Soc.  Exp.  Med.,  128(1):42-46.

Mackerer,  C. R., Saundefs, R. N., Haettinger, J. R., and Mehlman, M. A.,
     (1975) "Influence of 1,3-Butanediol on Blood Glucose Concentration and
     Pancreatic Insulin  Content of Streptozotocin-Diabetic Rats," Fed.
     Proc.,  34(12):2191-2196.

Maguire, M. P.,  (1974) "Chemically Induced Abnormal Chromosome Behavior at
     Meiosis in Maize," Chromosoma, 48:213-223.

Majchrowicz, E., Hunt, W. A., and Piantadosi, C.,  (1976) "Suppression by
     1,3-Butanediol of the Ethanol Withdrawal Syndrome in Rats," Science,
     194(4270):1181-1182.
                                    238

-------
Marcus, R. J., Winters, W. D., and Hultin, E.,  (1976) "Neuropharmacological
     Effects Induced by Butanol, 4-Hydroxybutyrate, 4-Mercaptobutyric Acid
     Thiolactone, Tetrahydrofuran, Pyrrolidine, 2-Deoxy-D-Glucose and
     Related Substances in the Rat," Neuropharmaco1., 13(l):29-38.

Martin, G. and Finberg, L.,  (1970) "Propylene Glycol:  A Potentially Toxic
     Vehicle in Liquid Dosage Form," J. Pediat., 77(5):877-878.

Marzulli, F. N. and Maibach, H. I.,  (1973) "Antimicrobials:  Experimental
     Contact Sensitization in Man,"  J.  Soc. Cosmet. Chem., 24:399-421.

Mason, M. M.,  (1971) "Toxicology and Carcinogenesis of Various Chemicals Used
     in the Preparation of Vaccines," Clin. Toxicol., 4(2):185-204.

Mattia, M. M., (1971) "Process for Recovering Organic Material From Aqueous
     Streams," U. S. Patent  3,625,886,  December 7, 1971, 4 pp.

Maxwell,  R. and Roth, R. H.,  (1972)  "Conversion of 1,4-Butanediol to Gamma-
     Hydroxybutyric Acid in Rat Brain and in Peripheral Tissue," Biochem.
     Pharmacol.,  21(11):1521-1533.

McChesney, E. W., Golberg, L., Parekh,  C. K., Russell, J.  C., and Min, B. H.,
     (1971) "Reappraisal of  the Toxicology of Ethylene Glycol.  II.
     Metabolism Studies in Laboratory Animals," Fd. Cosmet. Toxicol.,
     9:21-38.

McChesney, E. W.  and Golberg, L.,  (1972)  "Reappraisal of the Toxicology of
     Ethylene Glycol. IV.  The Metabolism of Labelled Glycollic and
     Glyoxylic Acids in the  Rhesus Monkey," Fd. Cosmet. Toxicol., 10:655-670.

McDonald, T. 0.,  Roberts, M.  D., and Borgmann, A. R., (1972) "Ocular Toxicity
     of Ethylene  Chlorohydrin and  Ethylene Glycol in Rabbit Eyes," Toxicol.
     Appl. Pharmacol.,  21(1):143-150.

McLaughlin, J., Marliac, J.,  Verrett, M.  Jr., Mutchler, M. K., and Fitzhugh,
     0. G.,  (1963) "The Injection  of Chemicals  into the Yolk Sac  of Fertile
     Eggs Prior to Incubation as a Toxicity Test," Toxicol. Pharmacol.,
     5:760-771.

MCP, (1976) "Chemical Product Synopsis:   Propylene Glycol," Mannsville
     Chemical Products, Mannsville,  N.Y.

MCP, (1977) "Chemical Product Synopsis:   Ethylene Glycol," Mannsville  Chemical
     Products, Mannsville, N.Y.

Mehlman,  M. A., Therriault,  D. G., Porter, W.,  Stoewsand,  G.  S.,  and  Dymsza,
     H. A.,  (1966) "Distribution of  Lipids in  Rats Fed  1,3-Butanediol,"  J.
     Nutr.,  88(2):215-218.

Mehlman,  M. A., Tobin,  R. B., and  Johnston, J.  B.,  (1970)  "Influence  of  Dietary
     1,3-Butanediol on  Metabolites and  Enzymes  Involved in Gluconeogenesis
     and  Lipogenesis  in Rats," J.  Nutr.,  100(11):1341-1346.
                                     239

-------
Mehlman, M. A., Tobin, R. B., Hahn, H. K. J., Kleager, L., and Tate, R.  L.,
     (1971a) "Metabolic Fate of 1,3-Butanediol in the Rat:  Liver  Tissue
     Slices Metabolism," J. Nutr., 101:1711-1718.

Mehlman, M. A., Tobin, R. B., and Johnston, J. B.,  (1971b) "Metabolic
     Control of Enzymes in Normal, Diabetic and Diabetic Insulin-Treated
     Rats Utilizing 1,3-Butanediol," Metab., 20:149-167.

Mehlman, M. A., Tobin, R. B., and Mackerer, C. R.,  (1975) "1,3-Butanediol
     Catabolism in the Rat," Fed. Proc., 34(12):2182-2185.

Menon, M. K.,  Clark, W. G., Schindler, A., and Alcarez, A., (1973)  "Studies
     on the Mechanism of the Akinetic Effect of Gamma-Hydroxybutyric Acid
     and 1,4-Butanediol," Arch. Int. Pharmacodyn.,  203:232-242.

Miller, P. H., (1966) "Glycols," Kirk-Othmer Eucycl. Chem. Technol.  (2nd
     Ed.), 10:638-660.

Miller, 0. N.  and Bazzano, G.,  (1965) "Propanediol Metabolism and  its
     Relation  to Lactic Acid Metabolism," Ann. N.Y. Acad. Sci., 119:957-973.

Miller, S. A.  and Dymsza, H. A.,  (1967) "Utilization by the Rat of  1,3-
     Butanediol as a Synthetic Source of Dietary Energy," J.  Nutr.,
     91(1):79-88.

Milles, G.,  (1946) "Ethylene Glycol Poisoning," Arch. Pathol., 41:631-638.

Morihara, K.,  (1965) "Production of Proteinase on Noncarbohydrate  Carbon
     Sources by Pseudomonas aeruginosa," Appl. Microbiol., 13(5):793-797.

Morizono, T. and Johnstone, B. M.,  (1975) "Ototoxicity of Chloramphenicol
     Ear Drops With Propylene Glycol as Solvent," Med. J. Aust., 2(18):634-638.

Morris, H. J., Nelson, A. A., and Calvery, H. D., (1942) "Observations on  the
     Chronic Toxicities of Propylene Glycol, Ethylene Glycol, Diethylene
     Glycol, Ethylene Glycol Monoethyl Ester, and Diethylene Glycol Mono-Ethyl
     Ether," J. Pharmacol. Exp. Therap., 74:266-273.

Myers,  V. S. and Usenik, E. A., (1969) "Propylene Glycol Intoxication  of
     Horses,"  J. Amer. Vet. Med. Assoc., 155(12):1841.

Neuberg, C.  and Gottschalk, A., (1925)  ["Physiological Behavior of  Acetoins,"]
     Biochem.  Zeitschrift., 162:484-487 (German).

Newman, H. W., Van Winkle, W., Kennedy, N. K., and  Motton, M. C.,  (1940)
     "Comparative Effects  of Propylene Glycol, Other Glycols, and  Alcohol
     on the  Liver Directly," J. Pharmacol. Exper. Therap., 68(1)-.194-200.

Optiz,  K.,  (1958)  ["Glycogenic  Effect of the Glycols,"] Arch. Exper. Pathol.
     Pharmak., 234:448-454  (German).

Page, I. H., (1927) "Ethylene Glycol - A Pharmacological Study," J. Pharmacol.
     Exp. Therap., 30:313-320.
                                     240

-------
Parker, M. M., (1972) "Metabolic Effects of 1,3-Butanediol," Unpublished
     M.S. Thesis, University of Nebraska, 61 pp.

Parry, M. F., and Wallach, R.,  (1974) "Ethylene Glycol Poisoning," Am. J.
     Med., 59(1):143-150.

Paterni, L., Dotta, F., and Sappa, A.,  (1956)  ["Intoxication of Ethylene
     Glycol in Albino Rats with Special Emphasis on Hematological Effects"]
     Folia Med., 39:242-259 (Italian).

Perl, W., Silverman, F., Delea, A. C., and Chinard, F. P.,  (1976)
     "Permeability of Dog Lung  Endothelium to  Sodium, Diols, Amides, and
     Water," Am. J. Physiol., 230(6):1708-1721.

Peterson, D. I., Peterson, J. E., Hardinge, M. G., and Wacker, W. E. C. ,
     (1963) "Experimental Treatment  of Ethylene Glycol Poisoning," JAMA,
     186(10):955-957.

Pevny, I. and Uhlich, M., (1975)  ["Allergy Against Components of Medical and
     Cosmetic External Preparations.  Polyethylene Glycol,  Propylene, Glycol
     Hexantriole"] Hautarzt, 26(5):252-254 (German).

Phillips, L. I., Steinberg, M., Maibach, H. J., and Akers,  W. A., (1972)
     "Comparison of Rabbit and  Human Skin Response to Certain Irritants,"
     Toxicol. Appl. Pharmacol., 21(3):369-382.

Pinheiro, A. J. R., Liska, B. J., and Parmelee, C. E., (1968) "Inhibitory
     Effect of Selected Organic Chemicals on Pseudomonas fragi " J. Dairy
     Sci., 51(2):223-224.

Fitter,  P.,  (1976) "Determination of Biological Degradability of Organic
     Substances," Water Res., 10:231-235.

Pons, C. A. and Custer, R. P.,  (1946) "Acute Ethylene Glycol Poisoning:  A
     Clinicopathologic Report of 18  Fatal Cases," Am. J. Med. Sci.,  211:544—552.

Portmann, J. E. and Wilson, K.  W.,  (1971) "The Toxicity of  140 Substances
     to  the Brown Shrimp and Other Marine Animals," Minist. of Agric., Fish.
     and Food, Fish. Lab., Burnham-on-Crouch,  Essex, England; Shellfish  Inf.
     Leafl., Number 22, 12 pp.

Price, K. S., Waggy, G. T., and Conway, R. A.,  (1974) "Brine Shrimp  Bioassay
     and Seawater BOD of Petrochemicals," J. Water Pollut.  Control  Fed.,
     46(l):63-77.

Rajagopal, G. and Ramakrishnan, S.,  (1975) "A  New Method  for Estimation  of
     Ethylene Glycol in Biological Material,"  Anal. Biochem., 65:132-136.

Rajagopal, G., Venkatesan, K.,  Ranganathan, P., and Ramakrishnan,  S.,  (1977)
     "Calcium and Phosphorus Metabolism in Ethylene Glycol  Toxicity in Rats,"
     Toxicol. Appl. Pharmacol., 39(3):543-547.
                                     241

-------
 Reid,  R.  W.  and Ivery,  D.,  (1975)  "A Rapid Gas Chromatography Procedure for
      Determining Glycols  in Serum,  Gastric and Urine Specimen," Clin. Chem.,
      21(7):1000.

 Reyniers,  J.  A.,  Sacksteder,  M.  R.,  and Ashburn,  L. L., (1964) "Multiple
      Tumors  in Female Germfree In-Bred Albino Mice Exposed to Bedding
      Treated  With Ethylene  Oxide,"  J.  Nat. Cancer Inst., 32(5):1045-1056.

 Richardson,  K.E., (1964)  "Effect of Testosterone  on Glycolic Acid Oxidase
      Levels  in Male and Female Rat  Liver,  . Endocrinol., 74:128-132.

 Richardson,  K. E., (1965)  "Endogenous Oxalate Synthesis in Male and Female
      Rats,"  Toxicol.  Appl.  Pharmacol., 7(4):507-515.

 Richardson, K.  E.,  (1973) "The Effect  of Partial  Hepatectomy on the Toxicity
      of Ethylene  Glycol, Glycolic Acid,  Glyoxylic Acid  and Glycine," Toxicol.
      Appl. Pharmacol.,  24:530-538.

 Roberts,J.A.  and  Seibold, H.R.,  (1969) "Ethylene  Glycol  Toxicity in
      the Monkey,"  Toxicol, Appl. Pharmacol., 15(3):  624-631.

Robertson, 0. H., Loosli,  C. G.,  Puck, T. T., Wise, H.,  Lemon, H. M., and
     Lester,  W., (1947) "Tests for the Chronic Toxicity of Propylene Glycol
     and Triethylene Glycol on Monkeys and Rats by Vapor Inhalation  and Oral
     Administration," J. Pharmacol.  Exp. Therap.,  91:52-76.

Romsos, D. R., Belo, P. S.,  and Leveille, G. A.,  (1974)  "Effect of 1,3-
     Butanediol on Hepatic Fatty Acid Synthesis and Metabolite Levels in the
     Rat," J. Nutr., 104(11):1438-1445.

Romsos, D. L., Belo, P. S.,  and Leveille, G. A.,  (1975)  "Butanediol  and Lipid
     Metabolism," Fed. Proc .,  34(12):2186-2190.

 Roth,R.H.  and Giarman^  N.J.,  (1968)  "Evidence  that Central Nervous  System
      Depression by 1,4-Butaned'iol is  Mediated  through a Metabolite,  Gammar-
      Hydroxybutyrate,"  Biochem.  Pharmacol.,  17(5):735-739.

 Rowe,  V.  K.,  (1963) "Glycols"  in Patty,  F.  A.  (ed.)  Industrial Hygiene  and
      Toxicology,  V.  II.,  Interscience Publishers, N.Y.,  pp.  1497-1536.

 Ruddick,  J. A., (1972)  "Toxicology, Metabolism, and Biochemistry of 1,2-
      Propanediol," Toxicol. Appl. Pharmacol.,  21(1):102-111.

 Rudney, H.,  (1954)  "Propanediol  Phosphates as  a Possible Intermediate in  the
      Metabolism of Acetone,"  J.  Biol.  Chem.,  210:361-371.
                        S
 Russell,  J.  C., McChesney,  E.  W., and Golberg, L.,  (1969)  "Reappraisal  of the
      Toxicology of Ethylene Glycol.  1.   Determination of Ethylene  Glycol
      in Biological Material by a Chemical  Method," Fd.  Cosmet. Toxicol.,
      7:107-113.

 Sax,  N. I.,  (1975)  Dangerous  Properties  of Industrial Materials, Van Nostrand
      Reinhold Co.,  N.Y.

 Scala, R.  A.  and  Paynter, 0.  E.,  (1967)  "Chronic  Oral Toxicity of 1,3-Butanediol,"
      Toxicol. Appl. Pharmacol.,  10:160-164.

                                     242

-------
Schliissel, H., (1954) ["Utilization of Polyvalent Alcohols in Nutrition"]
     Arch. Exper. Path. Pharmakol., 221:67-75 (German).

Schulz, M. and Comerton, L. J., (1974) "Effect of Aircraft Deicer on Airport
     Storm Runoff," J. Water Pollut. Contr. Fed., 46(1):173-180.

Schumacher, J. N., Green, C. R., Best, F. W., and Newell, M. P., (1977)
     'Smoke Composition:  An Extensive Investigation of the Water-Soluble Portion
     of Cigarette Smoke," J. Agric. Food Chem.,  25(2):310-20.

Seidenfeld, M. A. and Hanzlik, P. J.,  (1932) "The General Properties, Actions
     and Toxicity of Propylene Glycol," J. Pharmacol. Exp. Therap.,
     44:109-121.

Shanahan, B. W., (1977) "The Dermatopharmacologic Activity of Propylene
     Glycol in Selected Cosmetic Formulations," Diss. Abstr. Int. B.,
     38(4):1675.

Shelanski, M. V., (1974) "Evaluation of 1,3-Butylene Glycol as a Safe and
     Useful Ingredient in Cosmetics," Cosmet. Perfum.,  89(9):96,98.

Shumeyko, T. V. and Mansfield, R. G.,  (1975) "Properties and End Uses for
     Man-Made Fibers," in C. A. Harper (ed.). Handbook of Plastics and
     Elastomers, McGraw-Hill Book Co., N.Y., pp. 6-1 to 6-59.

Smith,  D.E., (1951)  "Morphologic Lesions Due to  Acute and Subacute Poison-
     ing with Antifreeze (Ethylene Glycol)," Arch.  Pathol.,  51:423.

Smyth, H. F., Seaton, J., and  Fischer, L.,  (1941) "The Single Dose Toxicity
     of  Some Glycols  and Derivatives," J.  Ind. Hyg.  Toxicol., 23:259-268.

Smyth, H. F., Carpenter, C. P., and Weil,  C. S., (1951)  "Range-Finding
     Toxicity Data:   List IV," Arch.  Ind.  Hyg. Occup. Med., 4:119-122.

Spitz, H. D.,  (1972)  "Ghosting of Ethylene Glycol in Gas Chromatography,"
     J.  Pharm. Sci.,  61(8):1339-1340.

Sprince, H., Josephs, J. A., and Wilptzeski, C. R.,  (1966)  "Neuropharmacological
     Effects of  1,4-Butanediol and Related Congeners Compared with  those of
     Gamma-Hydroxybutyrate  and Gamma-Butyrolactone," Life Sci.,  5(22):
     2041-2052.

SRI, (1975) Directory of Chemical Producers-USA, Stanford Research  Institute,
     Menlo Park, Calif..

Stenback, F. and Shubik, P.,  (1974)  "Lack  of Toxicity  and Carcinogenicity
     of  Some Commonly Used  Cutaneous  Agents," Toxicol. Appl. Pharmacol.,
     30:7-13.

Stoewsand, G.  S., Dymsza, H. A.,  Swift,  S. M., Mehlman,  M.  A.,  and  Therriault,
     D.  G.,  (1966)  "Effect  of  Feeding Polyhydric Alcohols on Tissue Lipids
     and the  Resistance  of  Rats  to Extreme Cold," J. Nutr.,  89:414-418.
                                     243

-------
Stoewsand, G. S. and Dymsza, H. A., (1967) "Synthetic Sources of Calories
     in the Diets of Rats and Dogs," Proc. Int.  Congr.  Nutr., 7th, 4:1082-1087.

Strack, E., Biesold, D., and Theile, H., (1960)  ["The Transformation and
     Distribution of 1,2-Propanediol,  1,2-Butanediol and 1,2,4-Butanetriol
     in Rabbits"] Zeit. Ges. Exp. Med., 132:522-537 (German).

Sunshine,  I.,  (1975) Methodology for Analytical Toxicology _, CRC Press,
     Cleveland,  Ohio,  ' pp.  163-166.


 Taberner, P. V- and Pearce, M. J., (1973) "Hypothermic  and Toxic Actions
      of 2-Butyne-l,4-diol and Other Related Diols in the Rat," J. Pharm.
      Pharmac., 26:597-604.

 Tanaka,K., Minamisawa, M., Manabe, M., and Matuura, S., (1975)  "Biological
      Test Using the Brine Shrimp:  Part 1. The Influence of Mycotoxins
      on the Brine Shrimp," Rep. Nat.  Food Res.  Inst. (Tokyo), 30:43-48.

Tanret,  P.,  Thomas, J., Thomas,  E., and Cottentot,  F.,  (1962)  ["Influence
     of  Sex  on  the  Formation of  Calcium Oxalate Crystals in  Rats  Intoxicated
     with Ethylene  Glycol"]  C.R.Soc. Biol., 156:1285-1287.

Tate,  R.  L., Mehlman,  M.  A.,  and Tobin, R.  B.,  (1971)  "Metabolic  Fate  of
     1,3-Butanediol in the  Rat:  Conversion to  g-Hydroxybutyrate,"  J.
     Nutrition,  101:1719-1726.

Taylor,  Ann,  (1976) "Two  Main Contenders  for  Ethylene Glycol Markets,"
     Chem. Age,  September 3,  1976:7.

Thomas,  J. F.,  Kesel,  R., and Hodge, H. C., (1949)  "Range-Finding Toxicity
     Tests on Propylene Glycol in  the  Rat," J.  Ind.  Hyg. Toxicol.,
     31:256-257.

Tobin,  R.  B., Mehlman,  M.,  and Parker,  M.,  (1972)  "Effect  of 1,3-Butanediol
     and Propionic  Acid on  Blood Ketones,  Lipids  and Metal  Ions  in  Rats,"
     J.  Nutr.,  102:1001-1008.

Tobin,  R.  B., Mehlman,  M. A.,  Kies, C., Fox,  H. M.,  and Soeldner, J.  S.,
      (1975)  "Nutritional  and Metabolic Studies  in Humans with 1,3-Butanediol,"
     Fed.  Proc., 34(12):2171-2176.

Train,  R. ,  (1975)  "Reconsideration of  Ethylene  Oxide,  Ethylene Glycol,
     Methyl  Amines  and Oxo  Chemicals Processes,"  Fed.  Reg.,  40(233):
     56436-56442.

Troisi,  F. M.,  (1950)  "Chronic Intoxication by  Ethylene Glycol Vapor,"
     Brit. J.  Industr.  Med.,  7:65-69.

Tsukamura, M.,  (1966)  "Utilization of  Glycols and Certain  Other  Carbohydrates
     by  Mycobacteria  as Sole  Carbon Sources,"  Amer. Rev.  Resp.  Dis.,
     94(5):796-798.
                                    244

-------
Tsukamura, M. and Tsukamura, S., (1968) "Differentiation of Mycobacteria
     by Susceptibility to Nitrite and Propylene Glycol," Amer. Rev. Resp.
     Dis., 98(3):505-506.

Umeda, M., (1957) "Production of Rat Sarcoma by Injections of Propylene
     Glycol Solution of p-Quinone," Gann (Tokyo), 48:139-144.

Union Carbide Corp., (1976) Material Safety Data Sheet;Ethylene Glycol,
     Chemicals and Plastics, N.Y.

Union Carbide Corp., (1978) Glycols, Chemicals and Plastics, N.Y., 72 pp.

U. S. Bureau of the Census, (1972-1976) "U. S. Imports for Consumption and
     General Imports," Report FT 246.

U. S. Bureau of the Census, (1972-1977) "U. S. Exports Schedule B Commodity
     Groupings," Vol. Ft., 610.

U. S. Int. Trade Comm., (1973-1978) "Synthetic Organic Chemicals, U. S.
     Production and Sales," U. S. Int. Trade Comm. Publ., 833.

U. S. Tariff Comm., (1968-1972) "Synthetic Organic Chemicals," Tariff
     Comm. Publ., 248.

Vaille, C., Debray, C., Martin, E., Souchard, M., and Roze, C., (1963)
      ["Experimental Renal Lithasis by Ethylene Glycol in Male and Female
     Rats"] Ann. Pharm. Franc., 21:111-116 (French).

Vaille, C., Debray, C., Roze, C., Souchard, M., and Martin, E., (1971)
      ["Hyperglycemic Action of Propylene Glycol,"] Ann. Pharm. Fr.,
     24(12):577-582 (French).

Van Oostrom, C. G. and Van Limborgh, J., (1976) "The Influence of Propanediol-
     1,3 on the Development of the Legs, Wings and Lower Beak of the
     Chicken Embryo," Acta Morphol. Neerl.-Scand., 14:319-329.

Van Winkle, W., (1941)  "Toxicity and Actions of Trimethylene Glycol," J.
     Pharm. Exp. Therap., 72:227-232.

Van Winkle, W., and Newman, H. W.,  (1941) "Further Results of Continued
     Administration of Propylene Glycol," Fd. Res., 6(5):509-516.

Vasquez, B. J., Martinez, J. L., Jensen, R. A., and McGaugh, J. L.,  (1977)
     "Amnestic Effects of Propylene Glycol in Mice," Proc. West. Pharmacol.
     Soc., 20:179-183. <

Vergnaud, P., (1950) "Process for the Manufacture of Butanediol and  Butanone,"
     U. S. Patent 2,529,061, November 7, 1950, 4pp.

Voigt, J. and Piatkowski, B., (1973)  ["Metabolism of Propylene Glycol  in  the
     Rumen of Cows,"] Arch. Tierernahrung, 23(4):323-327  (German).
                                    245

-------
Wagner, F. S., (1966) "1,3-Butylene Glycol," Kirk-Othmer Encycl. Chem.  Technol.,
     (2nd Ed.), 10:660-667.

Walker, N. E., (1967) "Distribution of Chemicals Injected Into Fertile  Eggs
     and Its Effect Upon Apparent Toxicity," Toxicol. Appl. Pharmacol.,
     10:290-299.

Wallenius, K. and Lekholm, U., (1973) "Oral Cancer in Rats Induced by the
     Water-Soluble Carcinogen 4-Nitrochinoline N-Oxide," Odont. Revy,
     24:39-48.

WARE Institute, Inc., (1969) "The Injection of Newborn Mice With Seven
     Chemical Adjuvants to Help Determine Their Safety in Use in Biologicals,"
     NTIS PB-195 153.

Warshaw, T. G. and Hermann, F., (1952) "Studies of Skin Reactions to
     Propylene Glycol," J. Invest. Dermat., 19:423-430.

Weast, R. C.,  (ed.),  (1973) Handbook of Chemistry and Physics, CRC Press,
     Cleveland, Ohio.

Weatherby, J. H. and  Haag, H. B., (1938) "Toxicity of Propylene Glycol,"
     Amer. Pharm. Assoc.,  27:466-471.

Weil,  C. S., Woodside, M.  D., Smyth, H. F., and Carpenter, C. P., (1971)
     "Results of Feeding Propylene Glycol in the Diet to Dogs for Two
     Years," Fd. Cosmet. Toxicol., 9:479-490.

Whittman, J. S. and Bawin, R. R., (1974) "Stimulation of Gluconeogenesis
     by Propylene Glycol in the Fasting Rat," Life Sci., 15:515-524.

Whitlock, G. P., Guerrant, N. B., and Dutcher, R. A., (1944) "Response  of
     Rats to Diets Containing Propylene Glycol and Glycerol," Proc. Soc.
     Exp. Biol. Med., 57:124-125.

Wiley, F. H., Hueper, W. C., and von Oettingen, W. F., (1936) "The
     Toxicity and Potential Dangers of Ethylene Glycol," J. Indus. Hyg.
     Toxicol., 18(2):123-126.

Wiley, F. H., Heuper, W. C., Bergen, D. S., and Blood, F. R.,  (1938)
     "Formation of Oxalic  Acid from Ethylene Glycol  and Related Solvents,"
     J. Ind. Hyg. Toxicol., 20:269-277.

Williams, R. T.,  (1959)^Detoxication Mechanisms;  The Metabolism and
     Detoxication of  Drugs, Toxic Substances and Other Organic Compounds,
     Chapman and Hall,  Ltd., London, pp. 69-87.

Wills, J. H., Cholakis, J- M., and Coulston, F.,  (1969) "The Effects of
     Ethylene Glycol  on the Formation of Urine by the Cat," Toxicol.
     Appl. Pharmacol.,  14(3):633-634.
                                    246

-------
Windholz, M. (ed.), (1976) The Merck Index, 9th Ed., Merck and Co., Inc.,
     Rahway, N. J.

Yagi, 0. and Yamada, K.,  (1969) "Studies on the Utilization of Petrochemicals
     by Microorganisms.   Part 1.  Production of Lactic Acid from 1,2-
     Propanediol by Arthrobacter oxydans," Agr. Biol. Chem.,  33(11):1587-1593.

Young, J. W. ,  (1975) "Use of 1,3-Butanediol for Lactation and Growth in
     Cattle,"  Fed. Proc., 34(12):2177-2181.

Zabik, J. E.,  van Dam, D. P., and Maickel, R.  P.,  (1974) "Pharmacological
     and Toxicological Studies on 1,4-Butanediol," Res. Comm. Chem. Pathol.
     Pharmacol., 8(1):83-90.

Zaroslinski, J. F., Browne, R. K., and Possley, L. H.,  (1971) "Propylene
     Glycol  as a Drug Solvent in Pharmacologic Studies," Toxicol. Appl.
     Pharmacol., 19(4):573-578.

Zeitoun, M.  A. and Mcllhenny, W. F.,  (1971) "Treatment  of Wastewater From  the
     Production of Polyhydric Organics," prepared  for EPA, NTIS PB-213 841.
                                     247

-------
                    CONCLUSIONS AND RECOMMENDATIONS



     The environmental impact of the title glycols cannot be adequately


assessed because only limited monitoring data are available.  Major


sources of environmental contamination by ethylene glycol and 1,2-


propanediol are likely the disposal of spent antifreeze and the runoff


of de-icing fluids.  No environmental monitoring data are available for


the butanediol isomers.  Studies would be desirable to assess the magnitude


of antifreeze and de-icer disposal on the environment and to monitor


other possible sources of contamination (ex. production and use facilities)


by the glycols.  There is no evidence to suggest that the glycols


xdlll persist or bioaccumulate in the environment.  A potential aquatic


impact of the glycols is the imposition of a high oxygen demand due to


rapid biological oxidation which would have an adverse effect on aquatic


organisms through dissolved oxygen depletion.  This problem should be


addressed in further studies.


     As reviewed in section V (Exposure and Effects Potential), the most


likely source of human contact with the glycols is by ingestion; inhalation


is not a significant source because of the low vapor pressure of the


glycols.  In most areas, adequate biological data exist for ethylene glycol,


1,2-propanediol, and 1,3-butanediol.  There is no evidence to suggest


that these compounds are carcinogenic.  In one study, ethylene oxide-


sterilized bedding resulted in hemorrhage and tumors in mice; ethylene


glycol, as a breakdown product of ethylene oxide, was implicated as a
                      c

causative factor but this question remains to be resolved.  Only sparse


biological data exist for the less commercially significant glycols:


1,3-propanediol, and 1,2- , 2,3- , and 1,4-butanediols.  However, 1,4-


butanediol should be considered for further study because of its increasing


use in plastics and resins.


                                    248

-------
                                 APPENDIX

                        Summary of Sources Employed


     References used in this report were selected from searches of auto-

mated information retrieval systems, indices, standard references works,

journals, books, etc.  Manufacturers, researchers, and federal and state

agencies, among others, were contacted directly.

     The following is a list of on-line systems searched:

     Agricola
     Biological Abstracts
     Cancerline
     Chemical Abstracts
     Dissertation Abstracts
     Food and Science and Technology Abstracts
     National Technical Information System
     PTS Market Abstracts
     Smithsonian Science Information Exchange
     Science Citation Index
     Toxline
     Toj&ack

Also, the Technical Information Center data base was searched by the Na-

tional Institute of Occupational Safety and Health.

     Manually searched indices included:

     Biological Abstracts (1957-1970}
     Chemical Abstracts (1957-1971)
     Excerpta Media
       Cancer (1953-1978)
       Pharmacology and Toxicology (1965-1978)
       Development Biology and Teratology (1965-1978)
       Environmental Health and Pollution Control (1972-1978)
       Occupational Health and Industrial Medicine (1971-1978)
     Index Medicus (1957-1978)

     Appropriate books"and compendia were examined and current journals

screened.  The literature search is considered complete through November

1978.
                                   249

-------
TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO. 2.
EPA 560/11-79-006
4. TITLE AND SUBTITLE
Investigation of Selected Potential Environmental
Contaminants: Ethylene Glycol, Propylene Glycols and
Butylene Glvcols 	
7. AUTHOR(S)
Lynne M. Miller
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Sciences Information Services Organization
Franklin Research Center
20th and Parkway
Philadelphia, PA 19103
12. SPONSORING AGENCY NAME AND ADDRESS
Office of Toxic Substances
U.S. Environmental Protection Agency
Washington, B.C. 20460
3. RECIPIENT'S ACCESSION NO.
5. REPORT DATE
Mav 1979
6. PERFORMING ORGANIZATION CODE
8. PERFORMING ORGANIZATION REPORT NO.
FRC 80G-C4807-01
10. PROGRAM ELEMENT NO.
11. CONTRACT/GRANT NO.
68-01-3893
13. TYPE OF REPORT AND PERIOD COVERED
Final
14. SPONSORING AGENCY CODE
560/11-79-006
15. SUPPLEMENTARY NOTES
16. ABSTRACT
This report reviews aspects of production, use, environmental exposure and
biological effects of ethyl ene glycol, two isomers of propylene glycol (1,2- and
1,3-propanediol) and four isomers of butylene glycol (1,3-, 1,4-, 2,3-, and 1,2-
butanediol) . Annual production of ethylene glycol is about 3.7 billion pounds
for use primarily in antifreeze and polyester fiber. About  0.5  billion pounds of
1,2-propanediol are produced per year for use in polyester  resins,  food,  pharm-
aceuticals,  and cellophane.  Annual domestic demand for  1,4-butanediol is about
0.2 billion  pounds for use in the production of tetrahydrofuran and acetylenic
chemicals.   The other title-glycols are of less importance  commercially.
     The major  source of environmental contamination by  ethylene glycol and
1,2-propanediol is likely from the disposal of spent antifreeze and de-icing
fluids.  However,  limited monitoring data make it difficult to  adequately assess
environmental exposure to the glycols.  The glycols are  capable of  being degraded
by a variety of acclimated and unacclimated soil, water, and  sewage microorganisms.
     In humans,  ethylene glycol intoxication, usually as a  result of accidental
ingestion of antifreeze, may 'result in nausea, hypertension,  tachycardia, cardio-
pulmonary failure,  renal impairment, coma and death.  1,2-Propanediol is a GRAS food
additive of  low toxicity.  1,3-Butanediol has been studied  as a source of dietary energy
Few studies  are available on 1,2- . 2.3- and 1.4-butane_diol  or  on T ,^-
17.
                                KEY WORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
                                              b.lDENTIFIERS/OPEN ENDED TERMS
                             COSATI Field/Group
ithylene Glycol
Propylene Glycol
Butylene Glycol
Environmental Engineering
legulations
Coxicology
                           Biological  and
                           Medical  Sciences
                           -biology
                           -clinical medicine
                           -toxicology
18. DISTRIBUTION STATEMENT
  Document is available to the public  through
  the National Technical Information Service,
  Springfield, Virginia  22151
19. SECURITY CLASS (ThisReport/
    Unclassified
21. NO. OF PAGES
     266
20. SECURITY CLASS (TMs page)
22. PRICE
EPA Form 2220-1 (Rev. 4-77)   PREVIOUS EDITION is OBSOLETE

-------