United Siates
Environmental Protection
Agency
Office of Water
Regulations and Standards
Criteria and Standards Division
Washington DC 20460
EPA 440,5-80-042
October '980
Ambient
Water Quality
Criteria for
2,4-dichlorophenol

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      AMBIENT WATER QUALITY CRITERIA FOR

            2,4-DICHLOROPHENOL
                 Prepared By
    U.S.  ENVIRONMENTAL PROTECTION AGENCY

  Office  of Water Regulations and Standards
       Criteria and Standards Division
              Washington, D.C.

    Office of Research and Development
Environmental Criteria and Assessment Office
              Cincinnati, Ohio

        Carcinogen Assessment Group
             Washington,  D.C.

    Environmental Research Laboratories
             Corvalis, Oregon
             Duluth,  Minnesota
           Gulf Breeze, Florida
        Narragansett, Rhode Island

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                              DISCLAIMER
     This  report  has  been reviewed by  the  Environmental  Criteria and
Assessment Office, U.S.  Environmental  Protection  Agency,  and approved
for publication.   Mention of trade names or commercial products does not
constitute endorsement or recommendation for use.
                          AVAILABILITY  NOTICE
      This  document is available  to  the public  through  the National
Technical Information Service, (NTIS), Springfield, Virginia  22161.
                                     11

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                               FOREWORD

    Section 304  (a)(l)  of  the Clean Water Act  of  1977 (P.L.  95-217),
requires the Administrator  of the Environmental Protection Agency to
publish  criteria for water  quality  accurately reflecting  the  latest
scientific knowledge on the  kind and extent of all identifiable effects
on  health  and welfare  which may be  expected  from  the  presence  of
pollutants in any body of water, including ground water.  Proposed water
quality criteria for the 65  toxic pollutants  listed  under  section 307
(a)(l)  of  the  Clean Water  Act were developed  and  a  notice  of  their
availability was published  for public comment on March 15,  1979 (44 FR
15926), July 25, 1979 (44 FR 43660),  and  October 1, 1979 (44 FR 56628).
This document  is a  revision of those  proposed  criteria based  upon  a
consideration of comments received from  other  Federal  Agencies,  State
agencies,  special  interest  groups,  and  individual  scientists.   The
criteria contained in this document replace any previously published EPA
criteria  for  the  65 pollutants.    This criterion  document  is  also
published in satisifaction  of paragraph 11 of the Settlement Agreement
in  Natural  Resources Defense Council, et.  al.  vs. Train,  8  ERC 2120
(D.D.C. 1976),  modified, 12 ERC 1833 (D.D.C.  1979).

    The term "water  quality criteria"  is used  in  two  sections  of the
Clean Water Act,  section 304  (a)(l) and section 303 (c)(2).  The term has
a different program  impact  in each section.   In section 304,  the term
represents a non-regulatory,  scientific  assessment of  ecological  ef-
fects.  The criteria  presented  in  this  publication  are such scientific
assessments.   Such  water  quality criteria  associated with  specific
stream uses when  adopted as  State  water quality standards under section
303 become  enforceable maximum acceptable  levels of  a pollutant  in
ambient waters.  The water  quality criteria adopted in the State water
quality standards could have the same numerical  limits as  the  criteria
developed under section  304.  However,  in many situations States may want
to adjust water quality criteria developed under section 304 to reflect
local   environmental  conditions   and  human   exposure   patterns  before
incorporation  into  water quality  standards.    It  is  not  until  their
adoption as part of the  State water quality standards that the  criteria
become regulatory.

    Guidelines to  assist the States  in  the modification  of  criteria
presented  in  this   document,  in   the development  of  water  quality
standards, and  in other water-related programs of this Agency, are being
developed by EPA.
                                    STEVEN SCHATZOW
                                    Deputy Assistant Administrator
                                    Office of Water Regulations and Standards
                                   111

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                              CRITERIA DOCUMENT
                              2,4-DICHLOROPHENOL
CRITERIA
                                 Aquatic  Life
    The available data for 2,4-dichlorophenol indicate that acute and chron-
ic  toxicity to  freshwater  aauatic life  occur  at concentrations  as low  as
2,020  and  365 ug/1,  respectively,  and  would occur  at lower  concentrations
among  species that  are more sensitive than  those  tested.   Mortality  to  early
life stages  of  one species of  fish occurs  at  concentrations as  low as  70
ug/1.
    Only one  test  has been  conducted  with  saltwater  organisms  and  2,4-di-
chlorophenol and  no statement can be made  concerning  acute  or chronic  tox-
icity.

                                 Human  Health
    For comparison  purposes,  two  approaches  were used to derive criterion
levels  for  2,4-dichlorophenol.    Based  on available toxicity  data,  for  the
protection of public  health,  the derived level  is  3.09 mg/1.  Using avail-
able organoleptic data, for controlling  undesirable taste  and  odor Qualities
of ambient water, the estimated  level  is 0.3 ug/1.   It should be recognized
that organoleptic data as a basis for establishing a water Quality criterion
have limitations  and  have no  demonstrated relationship to potential   adverse
human health effects.
                                      VI

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                              TABLE OF CONTENTS
Criteria Summary

Introduction                                                   A-1

Aquatic Life Toxicology                                        B-l
     Introduction                                              B-l
     Effects                                                   B-2
          Acute Toxicity                                       B-2
          Chronic Toxicity                                     B-3
          Plant Effects                                        B-3
          Residues                                             B-4
          Miscellaneous                                        B-4
          Summary                                              B-5
     Criterion                                                 B-5
     References                                                B-7

Mammalian Toxicology and Human Health Effects                  C-l
     Exposure                                                  C-l
          Ingestion from Water                                 C-l
          Ingestion from Food                                  C-9
          Inhalation                                           C-16
          Dermal                                               C-16
     Pharmacokinetics                                          C-17
          Absorption                                           C-17
          Distribution                                         C-17
          Metabolism                                           C-17
          Excretion                                            C-19
     Effects                                                   C-19
          Acute, Subacute and Chronic Toxicity                 C-19
          Synergism and/or Antagonism                          C-23
          Teratogenicity                                       C-24
          Mutagenicity                                         C-24
          Carcinogenicity                                      C-24
          Other Effects                                        C-29
     Criterion Formulation                                     C-31
          Current Levels of Exposure                           C-31
          Special Groups at Risk                               C-31
          Basis and Derivation of Criterion                    C-31
     References                                                C-35

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                                    ACKNOWLEDGEMENTS
Aquatic Life Toxicity:

    William A. Brungs, ERL-Narragansett
    U.S. Environmental Protection Agency
David J. Hansen, ERL-Gulf Breeze
U.S. Environmental  Protection Agency
Mammalian Toxicology and Human Health Effects

    Gary Osweiler (author)
    University of Missouri

    John F. Risher (doc. mgr.) ECAO-Cin
    U.S. Environmental Protection Agency

    Bonnie Smith (doc. mgr.) ECAO-Cin
    U.S. Environmental Protection Agency

    A. Wallace Hayes
    University of Mississippi Medical Center

    Steven D. Lutkenhoff, ECAO-Cin
    U.S. Environmental Protection Agency

    Jerry F.  Stara,  ECAO-Cin
    U.S. Environmental Protection Agency

    Philip J. Wirdzek, OWPS
    U.S. Environmental Protection Agency
Herbert Cornish
University of Michigan

Patrick Durkin
Syracuse Research Corporation

Terence M. Grady, ECAO-Cin
U.S. Environmental Protection Agency

Van Kozak
University of Wisconsin

Herbert Schumacher
National Center for Toxicological Research

William W. Sutton, EMSL-LV
U.S. Environmental Protection Agency

Gunther Zweig, OPP
U.S. Environmental Protection Agency
Technical Support Services Staff:  D.J. Reisman, M.A. Garlough, B.L. Zwayer,
P.A. Daunt, K.S. Edwards, T.A. Scandura, A.T. Pressley,  C.A. Cooper,
M.M. Denessen.

Clerical Staff:  C.A. Haynes, S.J. Faehr, L.A. Wade, D.  Jones, B.J. Bordicks,
B.J. Quesnell, C. Russom, R.  Rubinstein.
                                          w

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                                 INTRODUCTION

    2,4-Dichlorophenol  (2,4-DCP)  is  a  commercially  produced  substituted
phenol used entirely  in the  manufacture  of  industrial  and agricultural prod-
ucts.   As  an  intermediate  in  the  chemical  industry,  2,4-DCP is  utilized
principally as  the feedstock for  the  manufacture of  the  herbicide,  2,4-di-
chlorophenoxyacetic acid (2,4-0),  2,4-0  derivatives  (germicides,  soil  steri-
lants, etc.) and  certain methyl  compounds  used  in mothproofing, antiseptics,
and seed disinfectants.  2,4-DCP is  also reacted  with  benzene sulfonyl chlo-
ride  to  produce  miticides  or  further  chlorinated  to  pentachlorophenol,  a
wood preservative (U.S. EPA,  1975).
    2,4-Dichlorophenol is  a  colorless,  crystalline  solid  having  the  empiri-
cal  formula  C^ClgO,  a   molecular  weight   of  163.0  (Weast,   1975),   a
density of 1.383  at 60°F/25*C and  a vapor pressure of  1.0 mm Hg  at  53.0°C
(Sax, 1975).  The melting  point  of 2,4-DCP  is  45°C,  and the boiling point is
210"C at 760 mm Hg (Aly and Faust,  1965; Weast,  1975).
    2,4-DCP is slightly soluble  in  water at neutral  pH and dissolves  readily
in ethanol and  benzene (Kirk and Othmer,  1964).   2,4-DCP behaves as  a  weak
acid and is highly  soluble in  alkaline solutions, since it readily forms the
corresponding  alkaline salt.  The dissociation  constant  (pKj for  2,4-DCP
                                                              a
has been reported to  be  7.48 (Pearce and Simpkins,  1968).   Unlike  the mono-
chlorophenols,  2,4-DCP is  not  volatile from aeneous  alkaline solutions (Kirk
and Othmer, 1964).
    2,4-DCP is readily prepared  without benefit  of  a  catalyst  using  gaseous
chlorine and molten phenol  at  80  to  100'C.   Synthesis  is  also  attainable
through the chlorination of the monochlorophenols (Kirk and Othmer,  1972).
                                      A-l

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    It  has  been demonstrated  that  phenol  is  auite reactive to  chlorine  in
dilute aaueous solutions over a wide pH  range  (Carlson  and  Caple,  1975;  Mid-
daugh and Davis, 1976).
    Although  2,4-DCP  presently has no  direct  commercial application,  it  is
used  as an   important  chemical   intermediate,  and  it   is  synthesized  from
dilute  aaueous  solutions.   Its  identification as  a metabolic  intermediate
and degradation  product of various commercial  products by plants  (Kirk  and
Othmer,  1972),  microorganisms  (Kearney  and   Kaufman,  1972;  Steenson  and
Walker,  1957;  Bell,  1957, 1960;  Evans  and Smith,  1954; Fernley  and  Evans,
1959;  Loos,  et  al. 1967a,b;  Loos,  1969) and sunlight  (Aly  and  Faust,  1964;
Crosby and Tutass, 1966; Mitchell, 1961) has been well established.
    Numerous  studies  on the  microbial  degradation  of 2,4-DCP  have been con-
ducted,  revealing  degradation to yield  succinic  acid  (Alexander  and  Aleem,
1961;  Macrae,  et al.   1963;  Paris  and  Lewis, 1973;  Fernley  and  Evans,  1959;
Bell,  1957,  1960;  Bollag, et  al.  1968a,b; Macrae  and   Alexander,  1965;  Chu
and Kirsch,  1972;  Ingols,  et al. 1966;  Chapman,  1972;  Duxbury,  et al.  1970;
Loos, et al.  1967b; Tiedje, et al. 1969).
    Few  data  exist  regarding the persistence of  2,4-DCP in  the  environment.
2,4-DCP  is  slightly  soluble   in water,  while  its  alkaline salts  are readily
soluble  in  aaueous  solutions.   Its  low  vapor pressure  and  non-volatility
from  alkaline solutions would cause it  to  be  only slowly removed  from sur-
face  water  via  volatilization.   Studies  have  indicated  low   sorption  of
2,4-DCP  from  natural surface waters by various clays (Aly and Faust, 1964).
                                      A-2

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                                  REFERENCES







Alexander,  M.   and  M.J.H.  Aleem.   1961.   Effect  of  chemical  structure  on



microbial  decomposition  of aromatic  herbicides.   Jour.  Agric.  Food  Chem.



9: 44.







Aly,  O.M.  and  S.D.  Faust.  1964.   Removal  of 2,4-dichlorophenoxyacetic acid



and derivatives from natural waters.  Jour. Am. Water Works Assoc.  57:  221.







Bell,  G.R.   1957.  Morphological  and  biochemical  characteristics of a soil



bacterium  which  decomposes  2,4-dichlorophenoxyacetic   acid.    Can.   Jour.



Microoiol.  3:  821.







Bell,  G.R.   1960.  Studies  or  a  soil  Achromobacter which  degrades  2,4-di-



chlorophenoxyacetic acid.  Can.  Jour.  Microdiol.  6:  325.







Bollag,  J.M.,  et al.   1968a.   2,4-0  metabolism: Enzymatic  hydroxylation  of



chlorinated phenols.  Jour. Agric. Food Chem.  16:  826.







Bollag,  J.M.,   et  al.    1968b.   Enzymatic  degradation  of  cnlorocatechols.



Jour. Agric. Food Chem.   16: 829.








Carlson, R.M. and R. Caple.  1975.  Organo-chemical  Implications  of  Water  of



Water  Chlorination.   Iji:  Proc.  Conf.   Environ.   Impact  Water  Chlorination.



p. 73.
                                     A-3

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Chapman, P.J.   1972.  An  Outline  of  Reaction Sequences Used for the Bacteri-
al Degradation  of  Phenolic Compounds.  In:  Degradation  of  Synthetic  Organic
Molecules  in  the  Biosphere.   Proc. Conf.  Natl. Acao.  Sci.,  Washington,  D.C.
p. 17.

Chu,  J.P.  and  E.J.  Kirsch.   1972.   Metabolism  of  PCP by  axenic  bacterial
culture.  Appl. Microbiol.  23: 1033.

Crosby,  O.G.  and  H.O.  Tutass.   1966.   Photodecomposition  of  2,4-dichloro-
phenoxyacetic acid.  Jour. Agric. Food Chem.  14:  596.

Duxbury, J.M.,  et  al,   1970.   2,4-D  metabolism:  enzmatic conversion of chlo-
romaleylacetic  acid to succinic acid.  Agric. Food. Chem.  18: 199.

Evans,  W.C.  and B. Smith.  1954.  The  photochemical  inactivation  and micro-
bial metabolism of the  chlorophenoxyacetic  acid  heroicides.   Proc.  Biochem.
Soc. 571.

Fernley,  H.N.  and W.C.  Evans.    1959.   Metaoolism  of  2,4-dichlorophenoxy-
acetic  acid by  soil pseudomonas.  Proc. Biochem. Soc.  73:  228.

Ingols,  R.S.,  et  al.   1966.    Biological   activity  of halophenols.   Jour.
Water Pollut. Control Fed.  38: 629.

Kearney, P.C.  and  D.N.  Kaufman.   1972.  Microoial Degradation  of  Some Chlo-
rinated  Pesticides.   In:  Degradation of Synthetic  Organic  Molecules  in  the
Biosphere.  Proc. Conf.  Natl.  Acad. Sci., Washington, D.C.   p.166.
                                     A-4

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Kirk,  R.E.  ana  D.F.  Othmer.   1964.   Kirk-Othmer Encyclopedia  of Cnemical
Technology.  2nd ed.   Interscience Publishers, New York.

Kirk,  R.E.  and  D.F.  Othmer.   1972.   Kirk-Othmer Encyclopedia  of Chemical
Technology.  3rd ed.   Interscience Publishers, New York.

Loos,  M.A.  1969.   Phenoxyalkanoic  Acids.   Part  I.   _In_:  P.C.  Kearney and
D.D. Kaufman (eds.), Degradation of Herbicides.  Marcel Dekker, New York.

Loos,  M.A.,  et  al.   1967a.   Formation of  2,4-dichlorophenol  and  2,4-di-
chloroanisole from 2,4-0 by Arthrobacter sp.  Can.  Jour. Microbiol.  13: 691.

Loos,  M.H.,  et al.  1967b.  Phenoxyacetate herbicide  detoxication by bacte-
rial enzymes.  Jour. Agric. Food. Chem.  15: 858.

Macrae,  I.C.   and  M.  Alexander.   1965.   Microoial  degradation  of selected
pesticides in soil.  Jour. Agric. Food Chem.  13: 72.

Macrae,  I.C.,  et  al.   1963.   The  decomposition  of  4-(2,4-dichlorophenoxy-
butyric) acid by Flavobacterium sp.   Jour. Gen. Microbiol.   32: 69.

Midaaugh,  D.P.  and W.P.  Davis.   1976.   Impact of Chlorination  Processes  on
Marine Ecosystems.   In: Water  Quality  Criteria  Research of  the U.S. Environ.
Prot.  Agency.  EPA  Report  No.  600/3/-76-079.   Presented at 26th  Annu.  Meet.
Am. Inst. Biol. Sci., August 1975.
                                     A-5

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Mitchell, L.C.  1961.  The  effect  of  ultraviolet  light (2537A) on 141 pesti-
cide chemicals by paper chromatography.  Jour. Off. Analyt. Chem.  44: 643.

Paris, D.F. and D.L.  Lewis.   1973.   Chemical  and  microbial degradation of 10
selected pesticides in aquatic systems.  Residur Rev.  45: 95.

Pearce,  P.J.  and  R.J.J. Simpkins.   1968.   Acid strengths  of  some picric
acids.  Can. Jour. Chem.  46: 241.

Sax,  N.I.   1975.   Dangerous  properties  of  industrial  materials.   4th  ed.
Van Nostrand Rheinold Co., New York.

Steenson,  T.I.  and N.  Walker.  1957.   The  pathway of  breakdown of 2,4-di-
chloro- and 4-chloro-2-methyphenoxyacetate by  bacteria.   Jour.  Gen. Micro-
biol.  16: 146.

Tiedje,  J.  M., et  al.    1969.   2,4-D metabolism: pathway of degradation of
chlorocatechols by Arthrobacter sp.   Jour. Agric.  Food Chem.   17:  1021.

U.S.  EPA.   1975.   Significant organic products and organic chemical  manufac-
turing.  Washington,  O.C.

Weast, R.D.,  (ed.)   1975.  Handbook of Chemistry  and Physics.   55th  ed.   CRC
Press, Cleveland, Ohio.
                                      A-6

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Aquatic  Life Toxicology*



                                  INTRODUCTION

    2,4-Dichlorophenol  has been  widely used  as  a chemical  intermediate in

the manufacture  of herbicides, germicides, temporary  soil  sterilants, plant

growth  regulators, mothproofing  agents,  seed disinfectants,  miticides,  and

wood preservatives.   In  spite of this, there  are  only limited toxicity data

available  dealing with effects  of 2,4-dichlorophenol  on  freshwater aquatic

organisms.   Flavor-impairment studies  with  2,4-dichlorophenol   showed  that

flesh  tainting in fish  occurred at substantially  lower  concentrations  than

those  that  produced  other  adverse  effects  on plant,  fish,  and   invertebrate

species.  For  this reason,  flavor-impairment  information  may be  an important

consideration  in   deriving  the 2,4-dichlorophenol  criterion  for freshwater

aquatic  organisms.  Additional testing of  2,4-dichlorophenol  is  necessary to

meet the minimum  data  base requirement, detailed  in  the  guidelines,  for  the

derivation of  a  criterion.  This testing should  verify if flavor-impairment

of  fish  flesh  is,  indeed, the  most  sensitive  and important parameter  for

protecting the presence of and the uses of freshwater aquatic organisms.

    Only one test has been conducted  with  2,4-dichlorophenol and saltwater

organisms.
*The reader  is referred  to  the Guidelines  for Deriving Water  Quality Cri-
teria for  the  Protection of  Aquatic Life  and  Its Uses  in order  to  better
understand  the  following  discussion  and  recommendation.   The  following
tables contain the  appropriate  data that were  found  in the  literature,  and
at the bottom  of each  table  are calculations  for  deriving various measures
of toxicity as  described in the Guidelines.
                                      B-l

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                                    EFFECTS
Acute Toxicity
    The data  base  for freshwater invertebrate species  (Table  1)  consists of
two  static  tests  on  a single  cladoceran  species.  The 48-hour  LCgQ values
determined for Daphnia magna by  Kopperman,  et  al.  (1974)  and U.S. EPA (1978)
were  2,610  wg/l and  2,600  u9/l,  respectively.   The  LC5Q values  from  both
tests show good  reproducibility  of  results  between investigators.  Since all
the  invertebrate  acute  data available  for  2,4-dichlorophenol are  for  only
one species,  it  is impossible  to determine the relative sensitivity of Daph-
nia magna with that of other invertebrate species.
    Only two  acute values  dealing with  2,4-dichlorophenol effects  on fresh-
water fish species were  available (Table 1) and  only  one  of these tests was
conducted  using  flow-through   conditions   and   measured   concentrations.
Phipps, et  al.  (Manuscript)  calculated a  96-hour LC50  of  8,230  ug/l  for
fathead minnows.  This  is  more than  4  times higher  than the 96-hour  LCcQ
of 2,020  ug/1 for bluegills determined under  static  test  conditions  using
nominal concentrations (U.S. EPA, 1978).  Although differences in test meth-
ods make  comparisons  difficult,  it  appears bluegills  may be  slightly  more
sensitive to 2,4-dichlorophenol than are fathead minnows.
    The LCgg  values  for  Daphnia magna  fall between  the  LCgQ values  found
for bluegills  and  fathead minnows.   From  the few acute data  available,  it
appears that  there are no  large differences in the sensitivity  of fish and
invertebrate species to 2,4-dichlorophenol.
    The 96-hour  LC5Q  values  for chlorinated  phenols  and  bluegills  (U.S.
EPA,  1978) in this  and other criterion documents  are directly  related to the
degree of chlorination.  These values  decrease from 6,590  ug/1 for 2-chloro-
phenol and 3,830 wg/l for  4-chlorophenol  to 60  and 70 yg/1  for pentachlo-
                                      B-2

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rophenol.  Data for other species do not correlate as well.
Chronic Toxicity
    The  freshwater chronic  data base  for  2,4-dichlorophenol  consists  of a
single embryo-larval  test  (Holcombe,  et al. Manuscript)  conducted  with  fat-
head minnows.   The chronic  limits determined from this  study  (290-460 pg/1)
were based  on effects on  larval survival,  and  the  resulting  chronic value
was 365 ug/l  (Table 2).
    There appears  to  be a  moderate  difference  between  the  concentration  of
2,4-dichlorophenol  that  cause  acute  and chronic effects  on  fathead minnows.
The acute-chronic ratio for this species is 23 (Table 2).
    Data  from an  embryo  exposure and  an  additional  4-day larval  exposure
with three  species of  fishes  at two water hardnesses  (Birge, et  al. 1979)
will be discussed  in the miscellaneous section.
    Species mean  acute  and  chronic values  for  2,4-dichlorophenol  are listed
in Table 3.
Plant Effects
    The toxicity of 2,4-dichlorophenol  to  freshwater  aquatic plants does not
appear  important   in  the  derivation  of  a  criterion  since deleterious  on
plants  (Table 4)  occurred  only at  much   higher  concentrations  than  those
which produced  acute  toxic effects on  fish and invertebrate  species.  How-
ever, the knowledge that 2,4-dichlorophenol  is  not highly toxic  to  plants  is
important because  2,4-dichlorophenol  is  used in producing the  commonly  used
herbicide, 2,4-D  (2,4-dichlorophenoxyacetic acid).   Some  observed  toxic ef-
fects of 2,4-dichlorophenol on  plants were  the  complete  destruction of chlo-
rophyll  in  Chlorella  pyrenoidosa  at  100,000 Mg/l  (Huang and Gloyna,  1968)
and  a  50 percent  reduction in  chlorophyll  in  Lemna minor at 58,320  wg/1
(Blackman, et al.  1955).  Huang  and Gloyna  (1968)  also determined  that there
                                      B-3

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was a  56.4  percent reduction  of  photosynthetic oxygen production  in  Chlor-
ella pyrenoidosa after exposure to 50,000 ug/1 for 120 minutes.
Residues
    No measured  steady-state  bioconcentration factor  (BCF)  is  available  for
2,4-d i ch1oropheno1.
Miscellaneous
    Birge,  et al.  (1979)  determined  LC^Q  values  for   three  fish  species
after embryo  exposures  and  after  additional  4-day larval  exposures  at hard-
nesses of  50 and  200 mg/1  as  CaC03 (Table  5).   The LC50 values  after  the
4-day  larval  exposures  were 80 and  70  ug/1   for  rainbow  trout, 390 and  260
ug/1  for  goldfish, and  1,350 and 1,070  pg/1  for  channel catfish  at hard-
nesses does   not  substantially affect  toxicity,  although  for  all  species
tested the  LC^Q  values  for  2,4-dichlorophenol  were  slightly higher  at  low
hardness  than at  high  hardness.   The  LCcn  value of  260 ug/1 for  goldfish
(Birge, et al. 1979), which was based on  an  8-hour  total  exposure  of embryos
and  larvae,   was  considered  to be  the  lowest freshwater  acute  value  for
2,4-dichlorophenol.  Rainbow  trout embryos and  larvae exposed in  this study
were  fathead minnows   since  the  chronic value  for  fatheads is  365  ug/1
(Holcombe,  et al.  Manuscript).   Since  the   rainbow  trout  LCgQ  of 70  ug/1
(Birge, et al. 1979)  was based on a  relatively long-term study  (24-day  em-
bryo exposure plus a 4-day  larval  exposure),  this  value  was  considered to be
the lowest chronic value.
    Flavor-impairment studies  (Shumway  and Palensky,  1973) showed  that flesh
tainting  occurred  when  2,4-dichlorophenol  concentrations ranging from  0.4
ug/1  to  14  ug/l»  depending  on  the   species  of fish  tested, were  exceeded
(Table 5).    Based  on  the  available  data  for 2,4-dichlorophenol,  flavor-
impairment in fish occurs at  lower concentrations  than other effects used to
                                     B-4

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evaluate  toxicity  and may be  an  important consideration  in  deriving  a cri-
terion.   Since  the purpose of  the  Guidelines is  to  set water  quality cri-
teria which  protect  both the presence and  uses  of aquatic life, a criterion
which will  protect against tainting of  fish  flesh is necessary to preserve
the quality  of  the freshwater  fishery.   However,  the lack of  toxicity data
for 2,4-dichlorophenol makes  it impossible  to ascertain  if a  criterion based
on flavor-impairment of  fish  flesh  would be low  enough to protect all  fresh-
water aquatic organisms.
    The only saltwater  data  available  on the effects of 2,4-dichlorophenol
are  from  an acute exposure  to mountain bass, a  species endemic  in  Hawaii
(Hiatt, et  al.  1953).  Abnormal behavioral responses, including rapid swim-
ming in a vertical position,  gulping  at  the surface of  the water,  and jerky
motions, were observed in a nominal  concentration of 20,000 pg/1.
Summary
    Acute effects  on  freshwater fish and  invertebrate species  were observed
at concentrations from 260 to 8,230 pg/1.   The chronic value  for the fathead
minnow was 365  ug/1  with an  acute-chronic ratio of 23.   An embryo and 4-day
larval   exposure of rainbow  trout  yielded  an LC^Q  value of  70  yg/1.   The
lowest  plant effect  (50,000  gg/1)  caused  by  exposure to 2,4-dichlorophenol
was  based  on a reduction in  photosynthetic  oxygen  production in  an  algal
species.  Effects  on flavor-impairment of  largemouth  bass occurred when con-
centrations of 2,4-dichlorophenol  exceeded 0.4 yg/1.
                                   CRITERIA
    The  available  data  for  2,4-dichlorophenol   indicate  that  acute  and
chronic toxicity  to  freshwater aquatic  life  occur at concentrations  as  low
as 2,020 and 365 yg/1, respectively,  and would occur  at  lower concentrations
among species that are more sensitive than  those tested.   Mortality to early
                                      B-5

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life stages  of one  species  of fish  occurs  at  concentrations  as low  as  70
u9/1.
    Only one  test has been  conducted with  saltwater  organisms  and  2,4-di-
chlorophenol,  and no  statement  can  be  made  concerning acute  or  chronic
toxicity.
                                     B-6

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                       Table 1.  Acute values for 2,4-dlchlorophenol
LC50/EC50
Species Method* (fig/I)
Species Mean
Acute Value
(Ug/l) Reference
FRESHWATER SPECIES
Cladoceron, S, U 2,610
Daphnla magnn
Cladoceran, S, U 2,600
Oaphnla magna
Fathead minnow (juvenile), FT, M 8,230
Plmephales prone las
BluegHI, S, U 2,020
Lepomls macrochlrus

Kopperman, et al.
1974
2,605 U.S. EPA, 1978
6,230 Phlpps, et al.
Manuscript
2,020 U.S. EPA, 1978
* S B static, FT = flow-through, U c unmeasured, M » measured



  No Final Acute Values are calculable since the mini mum data base requirements are  not met.
                                               B-7

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Table 2.  Chronic valu*s for 2.4-dlchloropnwtol (Holcoatw, «t al. Manuscript)
     Sp«cU
     Fathead minnow,
     Plmephales prcreelas
           LlMltS
Method*    (iig/J)
                            FRESHWATER SPECIES
ELS
           290-460
                             M»an
                     Chronic ValiM
                        (Hfl/0
    J65
       ELS - early life stage


                            Acute-Chronic Ratio

                                 Chronic     Acute
                                  Value      Value
                                  (ug/l)     (U9/D
         Fathead minnow,
         Plreephales proroelas
365
           8,230
Ratio

 23
                                      B-8

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         Table 3.  Species Mean acute and chronic values for 2,4-dichlorophenol

                                        Specie* Mean     Species Mean
                                        Acute Value*     Chronic Value     Acute-Chronic
Number     Specie*                         (ua/l)           (yg/1)            Ratio**
3
2
1
FRESHWATER SPECIES
Fathead minnow, 8,230 365
Plmephales prone las
Cladoceran, 2,605
Daphnla magna
Blueglll, 2,020
Lepomls macrochlrus

23
-
-
* Rank from high concentration to  low concentration by species mean acute value.

**See the Guidelines for derivation of this ratio.
                                          B-9

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                 Table 4.  Plant values for 2.4-dlchlorophenol


                                                   Result
Species                            Effect          (pg/l)     Reference

                                FRESHWATER SPECIES
Alga,                          Complete           100,000      Huang  & Gloyna,
Chloreda pyrenoldosa          destruction of                   1968
                               chlorophy11

Alga,                          56.4$ reduction     50,000      Huang  & Gloyna,
Chloral la pyrenoldosa          of photosynthetlc               1968
                               oxygen production

Duckweed,                      50* reduction       58,320      Blackman,  et  al.
Lemna minor                    In chlorophyll                   1955
                                       B-10

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Table 5.  Other data for 2,4-dlctilorophwtol
Species
Duration
Effect
FRESHWATER SPECIES
Lymnaeld snails,
Pseudosucclnea columella
Fossarla cubensls
Crayfish,
Orconectes proplnquus
Orconectes inmunls
Cambarus robustus
Crayfish,
Orconectes proplnquus
Orconectes Imnunls
Cambarus robustus
Crayfish,
Orconectes proplnquus
Orconectes inmunls
Cambarus robustus
Crayfish,
Orconectes proplnquus
Orconectes inmunls
Cambarus robustus
Rainbow trout,
Sal mo galrdneri
Rainbow trout,
Sal mo galrdnerl
Rainbow trout,
Sal mo galrdnerl
Rainbow trout.
Sal mo galrdnerl
Rainbow trout.
Sal mo galrdneri
24 hrs
48 hrs
1 wk
10 days
10 days
48 hrs
24-day embryo
exposure
24-day embryo
exposure
24-day embryo
plus 4-day
larval exposure
24-day embryo
plus 4-day
100* mortality
100J mortality
1009 mortality
14 % mortality
Increased blood
glucose levels
ETC"
LC50 at hardness
of 50 mg/l CaC03
LC50 at hardness
of 200 mg/l CaC03
LC50 at hardness
of 50 mg/l CaC03
LC50 at hardness
of 200 mg/l CaC03
Result
(ug/D
10,000
10,000
5,000
1,000
1,000
1
80
70
80
70
Reference
Batte & Swanson,
1952
Tel ford, \974
Tel ford, 1974
Tel ford, 1974
Tel ford, 1974
Shumway & Palensky,
1973
Birge, at al. 1979
Blrge, et al. 1979
Blrge, et al. 1979
Blrge, et al. 1979
larval  exposure
                   B-ll

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Tab I* 5.  (Continued)
Species

Goldfish,
Cnrnsslus nuratus

Goldfish,
Carasslus aurntus

Goldfish,
Carasslus aurntus
Goldfish,
Carass I us auratus
Goldfish,
Carass Ius auratus

Fathead minnow
(juvenila),
Plmepholes promelas

Channel catfish,
Ictalurus punctatus

Channel catfish
Ictalurus punctatus

Channel catfish,
Ictalurus punctatus
Channel catfish,
Ictalurus punctatus
Blueglll,
Lepomls macrochlrus

Largemouth bass,
Mlcropterus SB I moIdes
   Duration

  4-day embryo
   exposure

  4-day embryo
   exposure

  4-day embryo
   plus 4-day
larval  exposure

  4-day embryo
   plus 4-day
larval  exposure

     24 hrs
    192  hrs
  4-day  embryo
    exposure

  4-day  embryo
    exposure

  4-day  embryo
   plus  4-day
larval exposure

  4-day  embryo
   plus  4-day
larval exposure

     48  hrs
     48  hrs
     Effect

LC50 at hardness
of 50 mg/I  CaCOj

LC30 at hardness
of 200 mg/I CaC03

LC50 at hardness
of 50 ing/1  CaC03


LC50 at hardness
of 200 mg/I CaC03


LC50
LC50
LC50 at hardness
of 50 mg/l CaC03

LC50 at hardness
of 200 mg/l CaC03

LC50 at hardness
of 30 ng/l CaC03


LC50 at hardness
Of 200 mg/l CaC03


ETC*


ETC*
Result
(ua/l)     Reference

 1,760     Blrge, et at. 1979


 1,240     Blrge, et al. 1979


   390     Blrge, et a I. 1979



   260     Blrge, et al. 1979
 7,600     Kobayashl, et al.
           1979

 6,500     Phlpps, et al.
           Manuscript
 1,850     Blrge, et al. 1979


 1,700     Blrge, et al. 1979


 1,350     Blrge, et al. 1979



 1,070     Blrge. et al. 1979
    14     Shumway & Palensky,
           1973

     0.4   Shumway & Palensky,
           1973
                                                B-12

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Table 5.  (Continued)


                                                               Result
Species                      Duration          Effect           ((ig/l)      Reference

                                       SALTWATER SPECIES
Mountain bass**               Acute       Moderate              20,000     Hlatt,  et a I.  1953
Kuhlla sandvIcons 15          response     reaction
* ETC = the highest estimated concentration of material  that  will  not  Impair  the flavor of the
  flesh of exposed fish.

••Endemic  In Hawaii
                                            B-13

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                                  REFERENCES

Batte, E.G. and  L.E.  Swanson.   1952.  Laboratory evaluation  of  organic  com-
pounds as molluscicides and ovicides.  II.  Jour. Parasitol.  38 65.

Birge, W. J., et  al.   1979.   Toxicity of organic chemicals to  embryo-larval
stages of fish.   EPA-560/11-79-007.  U.S. Environ. Prot. Agency.

Blackman,  G.E.,  et  al.    1955.   The  physiolgical   activity of  substituted
phenols.    I.   Relationships  between  chemical  structure   and  physiological
activity.  Arch.  Biochem. Biophys.  54: 45.

Hiatt, R.W., et al.   1953.  Effects  of chemicals on a schooling fish, Kuhlia
sandvicensis.   Biol. Bull.  104: 28

Holcombe, G.W.,  et al.   Effects  of phenol,  2,4-dimethylphenol,  2,4-dichlo-
rolphenol, and pentachlorophenol  on  embryo, larval,  and early-juvenile  fat-
head minnows (Pimephales promelas).  (Manuscript)

Huang, J.  and  E.F. Gloyna.   1968.  Effect of  organic  compounds  on  photo-
synthetic oxygenation.   I.   Chlorophyll destruction  and suppression  of  pho-
tosynthetic oxygen production.   Water Res.  2: 347.

Kobayashio, K.,  et  al.   1979.   Relation  between  toxicity and accumulation of
various chlorophenols in goldfish.  Bull. Jap. Soc.  Sci. Fish.  45: 173.
                                     B-14

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Kopperman,  H.L.,  et al.  1974.  Aaueous  chlorination  and ozonation studies.



I.  Structure-toxicity  correlations  of phenolic compounds  to  Daphnia magna.



Chem. Biol. Interact.  9: 245.







Phipps, G.L.,  et  al.  The  acute  toxicity of phenol  and substituted phenols



to the fathead minnow.  (Manuscript)







Shumway, D.L.  and J.R.  Palensky.   1973.   Impairment  of the flavor of fish by



water pollutants.   EPA-R3-73-010.  U.S. Environ. Prot. Agency.







Telford, M.   1974.   Blood  glucose  in  crayfish.  II.   Variations  induced by



artificial stress.  Comp. Biochem. Physiol.  48A: 555.







U.S.  EPA.   1978.    In-depth  studies  on health  and environmental  impacts of



selected water pollutants.    Contract  No.  68-01-4646.   U.S.  Environ.  Prot.



Agency.
                                     B-15

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Mammalian Toxicology and Human Health Effects
                              EXPOSURE
 Ingestion from Water
      Absorption of  2,4-dichlorophenol  (herein referred to as 2,4-
 DCP in this  document) by biological  tissues may occur upon exposure
 to 2,4-DCP  either  dissolved  in  water or associated with suspended
 matter or sediments  in  water.   Chlorophenols such  as  2,4-DCP are
 weak acids  that become  increasingly ionized under alkaline condi-
 tions; they  are mainly non-ionized at physiological pH.  These pro-
 perties, together with  the lipophilic nature of chlorophenols (in-
 cluding 2,4-DCP),  make  it  likely that  chlorophenols would  be ab-
 sorbed from the gastrointestinal tract.
      Sources of 2,4-DCP  in water may be diffuse  (e.g., agricultural
 runoff) or localized (e.g., point source pollution from manufactur-
 ing waste discharges).   Sidwell  (1971) verified  the  presence  of
 2,4-DCP  in  2,4-dichlorophenoxyacetic  acid  (2,4-D)  manufacturing
 wastes.  Over  a  7-month period, total chlorophenol content ranged
 from 68 mg/1 of waste to a high of  125 mg/1, with 2,4-DCP content
 ranging as high as 89 percent of the total.   Details of these find-
 ings are presented in Table 1.
      If certain assumptions  are made,  the  data of Sidwell (1971)
 can be  used to estimate human  exposure to  2,4-DCP  from drinking
 water obtained below a  point source  of  this  type.  Assuming (1)  an
 effluent with a 2,4-DCP  concentration of 125  mg/1  in water contain-
 ing 100,000  mg/1  of total  solids   (Table  1),  (2)  dilution  in the
 watercourse occurring to the point  where  an acceptable freshwater
 total  solids  concentration of  1,000 mg/1 is  reached,  and (3)  no
 removal of 2,4-DCP by water treatment,  drinking water will contain
                                C-l

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                                        TABLE 1
                  Industrial Plant Effluent Content of Chlorophenols'
Date Sampled
Total Solids (mg/1)
Temperature {°C)
PH
Chlorophenols (mg/1)
Phenol Type (%)b
Phenol-
2-chloro-
2,4-dichloro-
2,6-dichloro-
2,5-dichloro-
4-chloro-
2,4,6-tr ichloro-
2,4 , 5-tr ichloro-
25 Jan
6,960
12
7.5
68

3.4
2.9
73.6
9.9
trace
2.5
2.8
4.7
3 Mar
40,100
18
7.6
118

6.2
6.1
17.9
41.7
6.2
12.1
9.9
trace
21 Apr
76,320
21
7.4
125

1.7
trace
20.0
38.8
1.7
18.3
19.5
trace
28 May
104,860
28.5
7.4
112

24.8
trace
11.4
30.5
trace
20.0
13.3
trace
27 Aug
11,000
24
7.0
74

trace
trace
89.0
3.0
1.8
2.8
3.4
trace
Modified from Sidwell, 1971
Percent of total phenols present
                                        02

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2,4-DCP at  an  estimated concentration of 1.25 mg/1.  If the daily
water intake of a 70  kg  person is assumed to be 2  liters,  this would
result  in  a daily 2,4-DCP dosage  of 0.036 mq/kg body weight/day.
It should be emphasized that  this  dosage is a worst case exposure
level  that  would probably occur  only  in  drinking  water obtained
below a 2,4-DCP-contaminated point source  discharge.
     Sharpee  (1973)  noted  the presence of 2,4-DCP in soil treated
with 2,4-dichlorophenoxyacetic acid  (2,4-D).  This presence may be
accounted for  by the photolytic  or microbial breakdown of 2,4-D
which will  be  discussed  later  in  this document.   Walker   (1961)
studied the contamination of ground water by the  migration of waste
products from  the manufacture of chemicals  at  the  Rocky Mountain
Arsenal, Denver,  Colorado.  Walker reported  the  phytotoxic proper-
ties of water caused  by  either 2,4-D  or  an  unnamed "closely related
compound."  The  2,4-D type compounds were not a direct product of
the Arsenal operations,  but  rather were  apparently  the  result of
chemical reactions which occurred within basins  used for the stor-
age of effluents  from a variety of Arsenal operations.
     If diffuse  or point source  contamination of waters with 2,4-
DCP  is  in  fact   occurring,  the  dissipation  of  this  compound  in
aquatic environments becomes an important consideration.  The major
avenues of  2,4-DCP dissipation that have been  studied are microbial
degradation and photodecomposition.  Aly and Faust  (1964) examined
the dissipation of 2,4-DCP from natural lake waters at a buffered
pH of 7.  In aerated lake  waters,  with initial  2,4-DCP concentra-
tions of 100,  500, and  1,000  pg/1,  the percentages  of 2,4-DCP re-
maining at 9 days were 0, 0.34, and 46  respectively.  By contrast,
initial concentrations of 100,  500, and  1,000  yg/1 in unaerated and

                               C-3

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unbuffered  waters  resulted  in  percentages of  40,  51.6,  and 56,



respectively, remaining at 17 days.   Aly  and  Faust  concluded that



the persistence of chlorophenol would tend to increase at lower pH



and under anaerobic conditions that might result from the decompo-



sition of excessive organic matter.



     Ingols, et al. (1966)  studied the degradation of various chlo-



rophenols by activated sewage  sludge  and concluded that 2,4-DCP was



degraded more rapidly  by  activated systems with previous exposure



to chlorophenols than by those with no previous  exposure to chloro-



phenols.  when activated sludge  was exposed to 2,4-DCP at  levels of



100 mg/1 of  sludge/  75  percent  of the chemical  disappeared in two



days, and essentially  100  percent was gone in five days.  Hemmett



(1972) showed that  microorganisms  acclimated to  the herbicide 2,4-D



could degrade 2,4-DCP without a lag  period,  implying similar bio-



logical pathways for the two  compounds.



     When 2,4-DCP  was subjected  to  aeration  basin  treatment, the



initial concentration of 64 mg/1  dropped  to an  undetectable  level



within five days (Sidwell,  1971);  this was more  rapid than the rate



of degradation  observed for  2,4-DCP   in distilled water.   When an



aerated lagoon  alone was used, removal of  all chlorophenols varied



from 55 to 89 percent.  Overall,  removal  using both lagoon and sta-



bilization  ponds  ranged  from 87  to 94  percent.    Thus, natural



degradation of  2,4-dichlorophenol  may be  enhanced by proper appli-



cation of effluent waste management  principles.



     Dissipation of  dichlorophenols   also  occurs  through photode-



composition  in  aqueous  solutions.   Aly and Faust (1964) demonstrat-



ed  that  (1)  2,4-DCP  was  decomposed  by  ultraviolet   light,  and
                               C-4

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(2) the rate of  photolysis  in  distilled  water decreased as pH de-



creased.  Degradation of 50  percent of 2,4-DCP by ultraviolet light



was accomplished  in  two minutes at pH 9.0,  in  five  minutes at pH



7.0, and in 34 minutes at pH 4.0.  The studies  of Aly and Faust were



conducted  with  light of  wavelength 253.7  nm,  which  is  slightly



shorter than the natural ultraviolet radiation wavelength range of



292 to 400 nm.



     The  riboflavin-sensitized  dimerization of  2,4-dichlorophenol



to tetrachlorodiphenyl ethers, tetrachlorodihydroxy-biphenyls, and



other  products   was  reported  by Plimmer  and  Klingebiel   (1971).



Chlorinated dibenzo-p-dioxins, which could have resulted from ring



closure of  these  tetrachlorodiphenyl  ethers,  were not detected in



the products of  photolysis.   The  authors  speculated that failure to



detect chlorinated dibenzo-p-dioxins may have been due  to the rapid



photolytic breakdown of  those dioxins.  Rapid photolysis of chlori-



nated dibenzo-p-dioxins was confirmed by Crosby, et al. (1971).



     That 2,4-dichlorophenol can be formed as a photolytic product



of the  herbicides  2,4-D and nitrofen (2,4-dichlorophenyl p-nitro-



phenyl  ether)  in aqueous suspension  under sunlight  or simulated



sunlight,  has been noted by several  investigators,  including Aly



and Faust  (1964),  Zepp, et al.  (1975) ,  and Nakagawa  and  Crosby



(1974a,b).  Crosby and Tutass  (1966)  irradiated 2,4-0 with artifi-



cial light  (254  nm)  and natural sunlight and observed  that under



both conditions  2,4-dichlorophenol was  formed as a photolytic prod-



uct of 2,4-D and  was  further degraded to  4-chlorocatechol.  2,4-DCP



was photolabile,  with  50  percent being  lost in  five  minutes at



pH 7.0.
                               C-5

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     Microbial  decomposition of  2 ,4-dichlorophenol  in  soils and



aauatic environments  has  been  extensively  studied.  Alexander and



A^eem  (1961) found that when 80 yg 2,4-DCP/ml medium was  incubated



in the presence of Dunkirk silt loam and Mardin silt  loam,  2,4-DCP



was  not  detectable after nine  and  five days,  respectively.  When



these  soil  preparations were treated  with  sodium  azide,  no disap-



pearance of 2,4-DCP was noted,  supporting the role of microbiologi-



cal  processes  in  the  degradation.   Loos,  et al.  (1967) found that



extracts of Arthrobacter  sp.  contained enzymes capable of  dehalo-



genating 2,4-dichlorophenol.  Degradation was rapid, with 100 per-



cent chloride  release from 2,4-DCP occurring  after four hours of



incubation.  The  cells  responsible  for the dehalogenating  process



were active when  cultured on  the  herbicides 2,4-D or 2-methyl-4-



chlorophenoxyacetate  (MCPA).    Kearney, et al.   (1972)   found  no



tetrachlorodibenzo-p-dioxin  in  soils  treated with  up  to 1,000 ug



2,4-DCP/g.  Sharpee (1973) measured 2,4-DCP present in the  soil as



a  consequence  of  2,4-D  application  and  found  that  2,4-DCP did



appear in soil  treated with 2,4-D, but it did not persist as  long as



the  2,4-D.



     As  indicated earlier,  the principal  source  of 2,4-dichloro-



phenol in soils is believed  to  be  the  herbicide  2,4-D.  The  various



intermediates  (including  2,4-DCP)  in  the  microbial metabolism of



2,4-D have  been characterized by several investigators (Spokes and



Walker, 1974;  Loos, et  al.,  1967;  Bollag,  et al., 1968;  Evans, et



al.,  1971;  Paris  and  Lewis,  1973;  Ingols,  et al., 1966;  Alexander



and  Aleem,  1961).  Kearney and  Kaufman (1972)  have shown that the



organisms capable  of degrading  2,4-D to  2,4-DCP continue the degra-

-------
dation process  to catechol intermediates and  finally  to succinic
acid.  Thus,  the  herbicide 2,4-D is eventually  biodegraded to an
ecologically acceptable product, succinic acid.
     Recently,  attention  has  focused on  a  potential chlorophenol
source of  a more  ubiquitous  nature than herbicide  and pesticide
applications.   Both municipal  and  industrial wastewater are often
subjected to chlorination to achieve disinfection and deodorization
(Barnhart and Campbell, 1972) .   One result  of  chlorination is the
reaction of chlorine with phenol to produce chlorophenols,  some of
which  are  a  source  of obnoxious  odors  and/or  taste  (Dietz  and
Traud,  1978;  Barnhart  and  Campbell,   1972;  Burttschell,  et  al.
1959; Hoak, 1957).
     Phenol has been observed to be  quite reactive  with chlorine in
dilute aqueous  solutions.   This high reactivity of  phenol  is at-
tributed to  the ring-activating,  electron-releasing  properties of
the  -OH  functional group  (Barnhart and Campbell,  1972;  Morris,
1978).   Halogen  substitution  is  favored in the  ortho- and para-
positions (Burttschell, et al. 1959).
     Chlorination of phenols results in  a stepwise substitution of
the 2, 4, and 6  positions  of the  aromatic ring.  Barnhart and Camp-
bell (1972)  felt that  it was probable that chlorination  resulted in
a complex  mixture of chlorophenols.   Burttschell,  et  al.  (1959)
chlorinated 1  liter  of 20  mg  phenol/ml  solution containing 2 g/1
sodium bicarbonate with 40 mg chlorine and isolated 2,4-DCP as one
of three major chlorophenols.   Relative  chlorophenol content deter-
mined in this study was as follows:
                               C-7

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          Component                Percent of Product

          Phenol                        1-2
          2-Chlorophenol                2-5
          4-Chlorophenol                2-5
          2,4-Dichlorophenol            20
          2,6-Dichlorophenol            25
          2,4,6-Trichlorophenol        40-50

(Absolute amounts of chlorophenols were not reported.)


     Lee and  Morris  (1962)  verified  the  stepwise chlorination of

phenol to chlorophenolic compounds.   Furthermore, they determined

that the reaction rate and yield of 2,4-DCP is quite pH dependent,

with a decrease  in generation  time  and  an increase  in  yield for

2,4-DCP as pH increases from 7 to 9.  The  reactions occurred  at  ini-

tial chlorine concentrations of  1 ug/g and phenol  concentrations of

50 ng/g.

     Jolley (1973) indicated that development  of chlorinated organ-

ics, including chlorinated  phenols,  is retarded  in solutions  with

high  ammonia  concentrations.    In  a  later  study,  Jolley,  et al.

(1978) examined the chlorination of sewage waters  under conditions

simulating those  used  for  disinfection of sewage  effluents and/or

antifoulant treatment of cooling waters of electric power generat-

ing plants.  Over 50 chloro-organic constituents  were  separated in

each analysis  of  concentrated  sewage  effluent  chlorinated  in the

laboratory with 2.5 to 6 mg/1 concentrations  of chlorine.   Similar

studies were also done on two bodies  of lake  water, each receiving

effluent  from  a  coal-fired, electric-power generating plant.   Tn

the three systems  examined, Jolley,  et al.    (1978) detected mono-

chlorophenols at ng/g levels but found no dichlorophenols.
                               C-8

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     Glaze, et al. (1978)  identified trichlorophenols,  but  found no



dichlorophenols,  in superchlorinated municipal wastewaters.  Thus,



in contrast  to  laboratory demonstrations of DCP formation, recent



work under  conditions  simulating the natural  environment  has not



established  that  2,4-DCP  is a  significant  product resulting from



chlorination of phenol-containing waters.



Ingestion from Food



     Any  2,4-dichlorophenol  contamination of  food products  (non-



aquatic) would probably result from  use  of  the  herbicide  2,4-D.  As



stated earlier, 2,4-DCP is a possible contaminant of  2,4-D,  as well



as an intermediate compound  in the biological and ohotolytic degra-



dations of 2,4-D.



     Absorption of 2,4-DCP has been  reported for  some plant  species



(Isensee  and Jones,  1971).   Total DCP  content  in  oat  and  soybean



plants  increased  for approximately  three weeks when the soil con-



tained a DCP concentration of 0.07 ug/g. As  these  plants grew, the



total  tissue content  of DCP  remained  relatively constant but de-



creased as the plants  matured.   At  the  time of harvest, oats con-



tained 0.01 yg of  DCP  per  gram  of plant tissue,  and soybeans con-



tained 0.02 yg/g.  The 2,4-DCP  did  not  seem to concentrate in the



plant seeds.  DCP was not detected in the grain  of these oat plants,



and the soybean  seeds  contained only 1  to  2 percent  of the total



plant DCP.  No evidence was found of DCP translocation in soybeans



after foliar application.



     The  conversion  of 2,4-D to 2,4-DCP has been  demonstrated in



sunflowers, corn,  barley, strawberries,  and  kidney beans.   Steen,



et al.  (1974)  found  that 2,4-DCP residues  in  plants  treated with

-------
2,4-D herbicide  were from 20 to 100  times  lower than residues of



2,4-D.  Related studies by Sokolov,  et  al.  (1974)  involved applica-



tion  of  2,4-D to  rice  fields.   At  harvest,  rice grain contained



neither 2,4-D  nor  2,4-DCP,  even  though 2,4-DCP was present in the



rice  plant.    The  2,4-DCP content  in potato  tubers  treated with



2,4-D amounted to  less than 10 percent of the  total 2,4-D content.



     There  is  little information on  the  transfer of  2,4-D or its



degradation compounds to food  products of  animal  origin.  Mitchell,



et al. (1946)  provided evidence for  the gastrointestinal absorption



of 2,4-D  in lactating  dairy  cows  and detected  the  herbicide in



blood serum during a 106-day oral dosing  study.  However, 2,4-D was



not  detected   in  the  milk.   More   recently,  2,4-D was  not  found



(detection limit 0.1 ug/g) in bovine milk  following oral  doses of 5



ug 4-(2,4-dichlorophenoxybutyric) acid [4-(2,4-DB}] (Gutenmann, et



al.  1963a)  or  2,4-D  (Hutenmann,   et  al.  1963b)  or  50  ug  2,4-n



(Bache, et al. 1964a) per gram of feed.



     Clark, et al.  (1975)  studied  the tissue  distribution of 2,4-



DCP  in sheep  and cattle fed 2,4-D  at  the relatively  high concen-



trations of  300, 1,000, and  2,000  ug/g  of  feed.   Given that the



daily ration  is  approximately  3  percent  of the body weight,  these



feed concentrations are equivalent to approximate dosages of 9, 30,



and 60 mg/kg body weight, respectively.  The treated diets were fed



for  28 days,  and resulting concentrations of  2,4-D and 2,4-DCP in



several edible tissues were determined (Table 2).  with a detection



limit of 0.05  ug/g,  analysis  did not detect 2,4-DCP in the  fat or



muscle of cattle and  sheep, even at  the highest  dose levels.  How-



ever, the  kidney and liver were found to contain  large amounts of
                               C-10

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                             TABLE 2
              2,4-D and 2,4-DCP Residues (in mg/kg)
                  in  Sheep  and  Cattle  Fed 2,4-Da
2,4-D (Dose mg/kg body weight/day)
Sheep
Compound
2,4-D
2,4-DCP

2,4-D
2,4-DCP

2,4-D
2,4-DCP

2,4-D
2,4-DCP
60
0.06
< 0.05

0.10
£ 0.05

0.98
0.16

9.17
0.26
60b
Muscle
40.05 4
40.05 4
Fat
9
0.05
0.05

0.15 0.13
<0.05 4*0.05
Liver
0.29 <
0.15
Kidney
0.37
0.07

0.05
0.11

2.53
0.56
Cattle
30 60
*0.05 0.07

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2,4-DCP proportional  to  the dose given.   Furthermore,  when sheep



were withdrawn from 2,4-D for one week, measurable amounts of 2,4-



DCP were still detected.   Based on the limited evidence presented,



the liver appeared  to retain 2,4-DCP for  longer  periods  than did



the kidney.   The  data presented do  not allow accurate calculation



of a depletion  rate,  nor can the total  time  period  of  measurable



residues be ascertained.


                                                        6)
     Sherman, et al.  (1972) fed technical grade Nemacide^ [0-{2,4-



dichlorophenyl)-0,0-diethyl phosphorothioate] at 50, 100, 200, and



800 ug/g of feed to laying hens  for  55  weeks.  Analysis for 2,4-DCP



(a metabolite of Nemacide^  by gas-liquid chromatography (detection



limit 0.006  to  0.208  ug/g)  resulted in detection of 2,4-DCP resi-



dues in liver and yolk,  but  not in  muscle  or  fat.   Details of the



analytical findings are presented in Table  3.  As  in  the studies of



Clark,  et al.  (1975), there does  appear  to be some predilection of



2,4-DCP for  liver,  even  when formed by biotransformation from two



parent compounds.   In  the hens studied, liver 2,4-DCP concentration



decreased as  the  dosage  of Nemacide^ was  decreased.   The highest



mean liver level of 2,4-DCP found in hens was 0.56 ug/g,  identical



to a mean level of 0.56 ug/g found in the kidneys of cattle fed 300



ug 2,4-D/g of feed (the dosage level  that most closely approximates



possible field exposure)  by Clark and coworkers  (1975).   It should



be noted  that  the Sherman study was  strictly  a  laboratory study,



using conditions  that  are  not likely to  occur  in the field.  How-



ever, in a worst-case  exposure  situation,  the  consumption of 2,4-



DCP-contaminated  chicken liver  and  cattle  kidney  could   result in



approximately equivalent human  exposure.
                               C-12

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                                        TABLE 3
                2,4-Dichlorophenol Residues in Laying Hens fed Nemacide^
                 [O-(2,4-dichlorophenol)-0,O-diethyl phosphorothioate]a
Nemacide^ Feed
Concentration
    in ppm
  Days since
  Withdrawal *
from Nemacide
         Residues of 2,4-DCP (ppm)
         	mean (and range)	
                                                 Liver
                                                   Egg Yolk
     800
     200
     100
      50
       0
       5
       7
      10
      14
      21

       0
       7
      14
      21

       0
       7
      14
      21

       0
       7
      14
      21
0.47(0.14-0.68)

0.50(0.25-0.75)

0.27(0.06-0.48)
0.19(0.11-0.27)

0.38(0.31-0.44)
0.30(0.24-0.35)
0.14(0.10-0.18)
0.36(0.07-0.64)

0.26(0.25-0.26)
0.56
0.15( <0.05-0.33)
0.05

0.31(0.16-0.46)
0.18(0.14-0.22)
0.06( <0.05-0.11)
0.05
0.61(0.34-0.75)
0.27

<0.12
0.15(0.12-0.17)
0.15 «,0.12-0,21)
 Modified from Sherman, et al. 1972
                                          C-13

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     Bjerke, et  al.  (1972)  dosed dairy  cows  for three weeks with



2,4-D concentrations as high  as  1,000  mg/kg of diet.  2,4-DCP was



not found in the milk or cream of treated cows.



     The results of Clark,  et al.  (1975)  can be used  to calculate a



worst case  estimate for the  degree of  human  exposure to 2,4-DCP



from consumption  of contaminated meat.   Assuming  (1)  an average



forage yield of two tons (1,818 kg)  per  acre,  (2) the retention of



all herbicide on  the treated  plants,  (3) an application rate of 1



Ib (454  gins) of 2,4-D per acre, and  (4)  consumption by an  animal of



3 percent of  its body weight per  day  in forage,  then the dosage



delivered to an  animal  eating forage contaminated with this  level



of 2,4-D would be approximately 7 mg 2,4-DCP/kg  body weight.  This



amount  corresponds roughly  to  the lowest  dosage   (9  mg/kg body



weight)  that was administered to cattle  and sheep by Clark and his



colleagues.   Based  on the  results of Clark, et al.,  it is reason-



able to expect that cattle  fed a constant diet of forage contami-



nated with  2,4-D applied at commercial  rates would accumulate 2,4-



DCP concentrations  of  approximately  0.11 yg/g  of liver  and 0.56



yg/g of kidney.



     If a 70 kg person  consumed 0.5  kg of kidney daily  at  a 2,4-DCP



residue concentration of 560  yg/kg,   that  person  would be  consuming



approximately 280  ug of  2,4-OCP,  or 4.0 ug/kq body weight, daily.



If that person  also ingested 36 ug  2,4-DCP/kg body  weight/day in



contaminated drinking water,  as  calculated  from 2,4-DCP  levels in



water downstream  from a 2,4-DCP manufacturing  plant  (see  Ingestion



from Water  section), the resultant daily 2,4-dichlorophenol dosages



would total 40 yg/kg.   It  is therefore clear  that the water  would
                               C-14

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contribute  90  percent of this  highest  calculated daily dosage of
2,4-DCP.
     It  should  be emphasized that an exposure  level of 4 ug  2,4-
DCP/kg body weight/day is a  worst case example for food  intake.  It
would only  occur  if  a person (1) ate 0.5 kg of kidney per day and
(2) all  the kidney  consumed was contaminated  with 0.11  ug  2,4-
DCP/g.   This  contamination  level would  probably occur only  if the
cattle  were fed  a  constant diet  of 2,4-D-sprayed  forage,  since
experimental evidence  (Clark, et al.  1975; Zielinski and Fishbein,
1967) indicates  that levels of  2,4-DCP  in  animal tissue diminish
rapidly  following withdrawal of  2,4-D from the diet.
     A bioconcentration factor (BCF)  relates  the concentration of a
chemical  in aquatic  animals to  the  concentration in the water in
which they  live.   The steady-state BCFs  for  a lipid-soluble com-
pound in the tissues of various  aquatic animals seem to be propor-
tional  to  the  percent lipid in  the  tissue.   Thus, the per  capita
ingestion of a lipid-soluble chemical can be  estimated  from the per
capita  consumptions  of fish  and shellfish,   the  weighted average
percent  lipids of  consumed  fish and shellfish, and a steady-state
BCF for  the chemical.
     Data from a recent survey on fish and shellfish consumption in
the United  States were analyzed by SRI  International  (U.S. EPA,
1980).   These data were used to estimate that the oer caoita con-
sumption  of freshwater and  estuarine fish  and   shellfish  in  the
United  States  is 6.5  g/day  (Stephan, 1980).   In addition,  these
data were used with  data  on  the fat content  of the edible portion
of the  same species  to estimate that the weighted average percent
                               C-15

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lipids  for  consumed freshwater  and  estuarine  fish  and shellfish



is 3.0.



     No measured steady-state bioconcentration factor is available



for 2,4-dichlorophenol, but the equation "Log BCF =  (0.85 Log P) -



0.70" (Veith, et al. 1979) can  be  used  to estimate from the octa-



nol-water partition coefficient  (P)  the  BCF  for aquatic organisms



that contain about  7.6 percent  lipids  (Veith,  1980).  Based on an



average  log  P  value of 3.19  (Hansch and Leo,  1979),  the steady-



state bioconcentration factor  for  2 ,4-dichloroohenol is estimated



to be 103.  An adjustment factor of 3.0/7.6 = 0.395 can be used to



adjust  the estimated BCF  from  the  7.6  percent lipids on which the



equation  is  based  to  the  3.0 percent lipids  that  is the weighted



average  for consumed fish and shellfish.  Thus, the weighted aver-



age bioconcentration factor  for  2,4-dichloroohenol and the edible



portion  of all freshwater and estuarine aquatic organisms consumed



by Americans is calculated to be 103 x 0.395 = 40.7.



Inhalation



     There is  no  direct  evidence to  indicate  that humans are ex-



posed to significant  amounts of 2,4-DCP  through  inhalation.   Al-



though the compound is volatile, no quantitative studies of inhala-



tion  exposure  or  general environmental  contamination  have  been



found.



Dermal



     Dermal exposure to 2,4-DCP would most likely occur during the



manufacture, transport, or  handling  of  the compound.   Due to  its



lipophilic nature  and  low degree of  ionization at physiologic oH,



absorption of 2,4-DCP would be expected; however, no data relating



to dermal absorption have been  found.





                               C-16

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                         PHARMACOKINETICS



Absorption



     No information concerning the direct  absorption of 2,4-dichlo-



rophenol  in  humans or  animals was  found.    Toxicity data  to be



developed later confirm the existence of systemic toxicosis, indi-



cating that 2,4-DCP is absorbed by several routes.



     Because of  their  high  lipid  solubility and low  ionization at



physiological pH, dichlorophenols  would be  expected  to be readily



absorbed following ingestion.



Distribution



     No information was found concerning distribution  of 2,4-DCP in



man.  The previously  discussed (see  Ingestion  from  Food section)



animal studies  (Clark,  et al.  1975;  Sherman,  et al.  1972)  demon-



strated distribution  of  2,4-DCP  in  liver,  kidney,   and  egg   (see



Tables 2 and 3).



Metabolism



     The biotransformation  of 2,4-DCP in humans has  not been re-



ported.   No  information could  be found concerning the  metabolism of



2,4-DCP administered directly to experimental animals.  However, a



limited amount of information on the metabolism  of 2,4-DCP derived



from administration of gamma and beta benzene hexachloride (BHC) to


                                                        14
mice was reported by Kurihara  (1975).  Mice were given C  -labeled



gamma- or beta-BHC by  intraperitoneal injection,  and the appearance



of metabolites  in  the urine  was  monitored.    2,4-DCP and 2,4-DCP



conjugates were found and identified  primarily as glucuronides and



sulfates  (Table  4).    Administration of  gamma-BHC  resulted  in  a



majority of the 2,4-DCP being conjugated as  the  alucuroni^e, while



beta-BHC administration  resulted   in  a  greater  amount  of sulfate



                               C-17

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                         TABLE 4
         Urinary  2,4-DCP Metabolites of Benzene
                  Hexachloride in Mice3

BHC I some r
gamma-BHC
beta-BHC
Form of
Glucuronide
4-5%b
1-2%
2,4-DCP
Sulfate
0-1%
3%

Total
4-6%
4-5%
Source:  Kurihara, 1975
Percent of total metabolites
                           C-1B

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conjugate.  Assuming that the mouse biotransforms  2,4-DCP resulting



from endogenous metabolism in a manner similar to 2,4-DCP directly



administered, then sulfate and glucuronide conjugation appear  to be



major metabolic pathways.



Excretion



     2,4-D has  been found to be rapidly  excreted  in the urine of



mice (Zielinski and Fishbein, 1967), rats  (Khanna and Fang, 1966),



sheep (Clark, et al. 1964),  and  swine (Erne,  1966a,b)  under various



dosing conditions.   The  phenoxy  herbicide MCPA  was also rapidly



excreted in cattle urine (Bache, et al.  1964b).  However, excretion



data from  studies  using 2,4-DCP are not  extensive,  and no infor-



mation was found for 2,4-DCP excretion in man.



     Karapally, et  al.  (1973)  found that  when  rabbits were  given



radioactive  gamma-BHC,  2.5  percent of total  radioactivity in the



urine was due to 2,4-DCP.  Data presented did not allow for deter-



mination of body burden or half-life.   Shafik, et  al.  (1973) admin-



istered a daily dose of Nemacide-^  in peanut oil orally  to rats for



three days.   After  administration  of 1.6 mg Nemacide^, 67 percent



of that compound  appeared in urine as 2,4-DCP  within three  days.



With a  dosage of 0.16  mg Nemacide , 70  oercent  of  the pesticide



appeared as 2,4-DCP within 24 hours.  Work  cited earlier  (Kurihara,



1975) indicated the appearance  of  metabolites of 2,4-DCP in  urine



as a result of gamma- and beta-BHC administration (see  Table  4).



                             EFFECTS



Acute, Subacute, and Chronic Toxicity



     Farquharson,   et  al.  (1958)   indicated  that  the  toxicity of



chlorophenols tends to  increase as chlorination is increased.
                               C-19

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     The mechanism of toxic action for 2,4-DCP in mammalian systems
_iri vivo has not been well defined.  Limited ^ri vitro studies indi-
cate two potential actions.  2,4-DCP inhibits  oxidative phosphoryl-
ation in rat liver mitochondria  and rat  brain homogenates (Farqu-
harson,  et al.  1958; Mitsuda, et al.  1963).  According to Mitsuda,
et al.  (1963) ,  inhibitory  activity of chlorophenols  was roughly
correlated with  the dissociation  constant  of the  inhibitor.   In
addition, chlorine atoms on the ortho position weakened the activ-
ity of mono- and  dichlorophenols as oxidative inhibitors.  A con-
centration of 4.2  x 10"  M 2,4-DCP  inhibited oxidative phosphoryla-
tion by 50 percent in  rat  liver mitochondria.  By comparison, pen-
tachlorophenol was approximately 40 times more  active, and dinitro-
phenol was twice as active  in inhibiting oxidative phosphorylation.
     Stockdale and  Selwyn  (1971)  reported  observations suggesting
that the phenol-induced mediation  of  the passage of protons across
the inner mitochondrial membrane is sufficient to cause uncoupling
of oxidative phosphorylation.  They further noted  that phenols have
direct effects on  the enzyme ATPase as well as on one or more compo-
nents of  the  electron transport system;  however,  neither of these
effects is actually  involved in  the uncoupling process.
     Farquharson,  et al. (1958)  offered the conclusion that chloro-
phenols with pK values of 7.85  or less appear  to be  acutely associ-
ated with  production of marked hypotonia and early onset of rigor
mortis after death.   Similar clinical effects are associated with
well known oxidative uncouplers, such as  2,4-dinitrophenol and pen-
tachlorophenol.   Relatively few studies of  the  acute  or subacute
toxicity  of  2,4-DCP  have  been  reported.   The acute  LD5Q values
determined by several  investigators are presented in Table 5.

                               C-20

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                                      TABLE 5

                        Acute Mammalian Toxicity of 2,4-DCP
Species
   Route of
Administration
 LD50
(mg/kg)
       Reference
 Rat

 Rat

 Rat

 Rat

 Mouse
Oral

Subcutaneous

Intraperitoneal

Oral

Oral
   580

 1,730

   430

 4,000

 1,600
Deichmann, 1943

Deichmann, 1943

Farquharson, et al. 1958

Kobayashi, et al. 1972

Kobayashi, et al. 1972
                                     C-21

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     In the study by Farquharson,  et al.  (1958) ,  acute poisoning of



rats following  intraperitoneal 2,4-DCP  injection appeared  to be



characterized by the onset of hypotonia two to three minutes after



dosing.  This effect began in  the  hindlimbs and moved forward until



the rats were prostrate.  Eye reflexes were weakened and there was



no withdrawal  from  toe  pinch.    Muscle  twitches  rarely occurred



spontaneously and could not be evoked by  auditory or tactile stimu-



li.  Rectal temperature was only slightly decreased.  Initial dose-



induced  polypnea  was  followed  by  slowed  respiration  and dyspnea



as coma ensued.  Rigor mortis appeared earlier in rats killed  with



2,4-DCP than in control rats killed with ether.



     The oral LDc0 derived by  Deichmann  (1943)  appears at odds  with



the findings of Kobayashi, et al.   (1972).  Deichmann used fuel oil



as a solvent, which may have enhanced rapid uptake of 2,4-DCP.  The



vehicle  for  the  Kobayashi studies could  not  be determined.   None-



theless, from the  LD5Q  values,  it appears  that 2,4-DCP would  con-



stitute an acute hazard only following massive exposure.



     In a  subacute  (10-day) study,  Kobayashi,  et al.  (1972)  found



that all mice  survived when 2,4-DCP  at  667  mg/kg body weight was



given orally.  They  derived LDcg  figures for this study which ap-



pear similar to  those  listed  for  the acute studies (see Table 5).



In the same study (Kobayashi,  et al. 1972), male  mice were also fed



2,4-DCP  in the  diet over  a 6-month period.   Parameters evaluated



included average body weight,  food  consumption, orqan weight,  glu-



tamic  oxaloacetate  transaminase,   glutamic  pyruvic  transaminase,



erythrocyte  counts,   leucocyte   counts,   and   histopathological



changes.   The  estimated dose  levels were 45  mg/kg/day, 100 mq/kg/
                               C-22

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day, and 230 mg/kg/day, corresponding to dietary  2,4-DCP concentra-
tions of  50C,  1,000,  and  2,000  ug/g,  respectively.   No adverse
effects were noted  in mice  at any  dose  level, except  for some
microscopic  non-specific  liver  changes after  the  maximum  dose.
These changes included infiltration of  round cells and swelling of
hepatocytes, with some differences in cell size.  Two animals were
reported  to have  "dark  cells"  in the  liver.    (To  the  author's
knowledge, this term is not  commonly  used in  the  United States, and
its meaning  is not  clear.)  Kobayashi,   et al.  concluded  that 100
mg/kg/day is a maximum no-effect level  in mice.
     No other  chronic  toxicity studies using  2,4-dichloroohenol
have been found.   One report in  the  literature  (Bleiberg,  et al.
1964)  has suggested  a possible  role  of  2,4-DCP  in acquired chlor-
acne and porphyria in workers manufacturing  2,4-DCP and 2,4,5-tri-
chlorophenol (2,4,5-TCP).   The  workers   involved were also exposed
to acetic acid, phenol, monochloroacetic acid,  and sodium hydrox-
ide.  Since  various  dioxins (including  2,3,7,8-tetrachloro-diben-
zo-p-dioxin  (TCDD), which has been associated with chloracne) have
been implicated as  contaminants of 2,4,5-TCP,  the role of 2,4-DCP
in  inducing  chloracne and  porphyria  is not conclusive  (Huff and
Wassom, 1974).
Synergism and/or Antagonism
     Reports of studies directly assessing  the  synergism or antago-
nism of 2,4-dichlorophenol  by  other   compounds  were not  found.
Since 2,4-DCP  is  an  uncoupler  of  oxidative  phosphorylation  (Mit-
suda,  et al.   1963),  it may be  expected that concomitant exposure
to other uncouplers  (e.g., pentachlorophenol, dinitrophenol) would
                               C-23

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enhance that effect.  In addition, exposure to gamma- or beta-BHC,



2,4-D, and nitrofen could add  slightly  to any primary body burden



of 2,4-DCP.



     Any agent causing liver damage sufficient to decrease  the con-



jugation of 2,4-DCP with glucuronide  or  sulfate could conceivably



alter the  excretion and/or  toxicity of  the oarent compound.  How-



ever, there are no specific studies to reflect such an effect.



Teratogenicity



     Pertinent data could  not  be located in the available  litera-



ture concerning the teratogenicity of 2,4-DCP.



Mutagenicity



     No studies were found which addressed the mutagenicitv of 2,4-



dichlorophenol in mammalian systems.  Amer and Ali  (1968, 1969) did



report  some  effects of  2,4-DCP on mitosis and  meiosis in flower



buds and root cells of vetch (Vicia faba).  Changes  included meiot-



ic alterations of  chromosome stickiness, lagging chromosomes, and



anaohase bridges when flower buds were sprayed with 0.1 M  2,4-DCP.



Mitotic changes of chromosome stickiness, lagging chromosomes, dis-



integration, bridging, disturbed proohase and metaphase, and occa-



sional cytomyxis were seen  in  root cells  exoosed  to 62.5 mg/1 DCP.



Later studies  (Amer and Ali, 1974)  further confirmed  the effect of



chromosome  stickiness,  lagging  chromosomes,  and fragmentation in



35-day-old Vicia faba.  The  relationship of  these changes to alter-



ations  in mammalian cells has  not been established.



Carcinogenicity



     Repeated  application  of  ohenol  and some substituted  ohenols



has demonstrated promoting,  as  well as complete,  tumorigenic activ-
                               C-24

-------
ity  (Boutwell and Bosch,  1959).   In  the Boutwell and Bosch study,

two trials  included  evaluation of 2,4-DCP as a  promoter.   In one

trial, 25  pi  of a 20  percent  solution of 2,4-DCP  in  benzene was

applied twice  weekly for 15  weeks to  female  Sutter mice  two to

three months  of age.  The other  trial  was  identical,  except that

2,4-DCP was applied  for 24 weeks.  Application in both trials fol-

lowed  an  initiating dose  of   0.3  percent dimethyl-benzanthracene

(DMBA) in benzene.

     The  2,4-DCP  dose used corresponds  to  5  mg of compound per

mouse at each  application, or  10  mg/week when applied twice weekly.

Sutter mice two to three  months  old  would be expected to weigh 35

grams, so  that  the  dose  rate would  be  40.82 mg/kg  body weight.

Tumorigenic response was measured as  follows:


     1)   The percentage of surviving mice bearing one or more
          papilloma was ascertained.

     2)   The number of  papillomas on all surviving mice was
          totaled and  divided  by the  number  of  survivors to
          give the average number of  papillomas per  surviving
          mouse.

     3)   The  number of  mice  bearing  malignant  tumors was
          determined.


     Results of  the  promoter   trial with  2,4-DCP are presented in

Table 6.   Related  promoter experiments with phenol and the two ben-

zene controls are included for comparative purposes.  Boutwell and

Bosch concluded that the  promoting  activity  of  2,4-DCP is similar

to  that  of phenol.   However,  no statistical analyses  nor dose-

response data were included to support this comparison.

     To see if there  was a  statistically  significant difference

between the 2,4-DCP-treated mice  and  the benzene  controls, a Fisher
                               C-25

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                                                        TABLE 6
                     Appearance of Skin Tumors in Mice Treated Cutaneoualy with Phenols following
                          a Cutaneous Dose of 0.3% Dimethyl-benzanthracene  (DMBA)  in Acetone9
Treatments
Benzene control
Benzene control
10% phenol in
benzene in DMBA
20% phenol in
acetone
20% phenol in
benzene
20% 2,4-DCP
in benzene
20% 2,4-DCP
in benzene
Time Animals
Examined
(week)
15
24
20
12
24
15
24
No. of Mice
(survivors/total)
15/20
27/32
24/30
21/24
10/33
27/33
16/23
Survivors
with PapilloMS
7
11
33
58
100
48
75
Average
Papilloaas
per Survivor
0.07
0.15
0.62
-
3.20
1.07
1.62
Survivors with
Epithelial Carcinomas
0
0
13
5
20
11
6







Source:  Modified from Boutwe11 and Bosch,  1959
All received DMBA except where stated
                                                  C-26

-------
exact test  was  undertaken  for  this document.  Tumor incidence was
derived with the assumption  that  only  survivors  were examined for
tumors.  The calculated results are presented in Table 7.
     This  analysis  indicates  that  (1)  the  higher  incidence  of
papillomas  in both 2,4-DCP-treated groups was not attributable to
chance, and (2)  the carcinoma incidence was not significantly ele-
vated over controls.
     This statistical exercise should not obscure a number of con-
siderations that could  affect  the meaningfulness  of  the results.
(1) The study used dermal application of a phenolic compound at 20
percent concentration in organic  solvents.   This concentration is
high enough to  destroy hair follicles and  sebaceous  glands.   The
papillomatous response observed may have  developed in  response to
chemical  and/or  physical damage  from application of  an irritant
compound.   (2)  Even with the  harsh treatment,  no  malignant  neo-
plasia was  observed,  except when DMBA was  used  as  an  initiator.
The only neoplasia observed was at the site of direct application.
(3) Pathological identification of benign and malignant  tumors was
done on a gross  level,  with only  periodic  confirmation  by micro-
scopic examination.   (4) The mice were housed in creosote-treated
wooden cages,  which themselves were capable of initiating a carci-
nogenic response.
     The report of Boutwell and Bosch (1959) is the only one found
that deals with the tumorigenicity of 2,4-DCP.  However, since the
study was  designed  primarily  to  detect  promoting  activity,  the
effect of 2,4-DCP as a primary carcinogen is not well defined.
                               C-27

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                  TABLE  7
Results of Fisher Exact Test Applied to Data
       from Boutwell and Bosch (1959)
Treatment „, ^ration of
Treatment (wks)
Benzene control (I) 15
20% 2,4-DCP 15
in benzene
Benzene control (II) 24
20% 2,4-DCP in 24
benzene
Incidence of P-value I£cid.e"S:e ?f P-value
Danii i^mae, vs- Epithelial vs.
fapiiiomas control Carcinoma Control
1/15
13/27 0.61 x 10~2
3/27
12/16 0.36 x 10"4
0/15
3/27
0/27
1/16
-
0.2548
-
0.3721
                C-28

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     The route  of  administration is  not  approoriate to the model



for carcinogenic risk assessment  and  has  no established relation-



ship to  oral  exposure.   Overall, the study does present evidence



that 2,4-DCP may be a possible promotor, and the  work may be appli-



cable to evaluating  the  hazard of skin or respiratory exposure to



2,4-DCP, alone or concurrent with other chemicals.



Other Effects



     An odor threshold for 2,4-DCP in water  has been  reported by at



least three investigators.   Hoak  (1957) determined the odor thresh-



old of 2,4-DCP to be 0.65 yg/1 at  30°C and 6.5 yg/1 at 60°C.  Deter-



mination of the detectable odor was made by a  panel of two or four



people  comparing  flasks  of  test water  to  a flask of  odor-free



water.   The lowest concentration  detected by any panel member was



taken as  the odor  threshold.   Hoak  speculated  that  odor  should



become more noticeable as temperature  increases;  however, in evalu-



ating a series of chlorophenols and  cresols, it was found that some



compounds had higher odor thresholds at 30°C,  and others were high-



er at 60°C.



     Burttschell, et  al.  (1959)  made dilutions of chlorophenol in



carbon-filtered tap water and used a panel of four to six people to



evaluate odor.   Tests were  carried  out at room  temperature, which



the investigator  estimated  to be 25°C.   If a  panel  member's re-



sponse was doubtful,  the sample was considered negative.  The geo-



metric mean of the panel responses was used as the odor threshold.



For 2,4-DCP the threshold was  2 ug/1.



     Dietz  and  Traud  (1978)   used  a  panel  composed  of  9  to  12



persons of both sexes and various age  groups to test  the organolep-
                               C-29

-------
tic detection  thresholds  for  126  phenolic compounds.   TO test for
odor thresholds,  200  ml  samples of  the  different  test concentra-
tions were placed in stoppered odor-free glass bottles, shaken for
approximately  five  minutes,  and sniffed  at  room temperature  (20-
22°C).   For each test,  water  without the  phenolic additive was  used
as a background sample.  The  odor  tests took  place  in several indi-
vidual  rooms  in which phenols  and other  substances  with intense
odors had  not  been used  previously.   Geometric mean  values were
used to determine  threshold  levels.  To determine taste threshold
concentrations of selected phenolic compounds,  a  panel  of four  test
individuals tested water samples containing various amounts of  ohe-
nolic additives.  As a point of comparison, water without phenolic
additives was  tasted first.  Samples with  increasing phenolic  con-
centrations  were then  tested.    Between  samples,  the  mouth  was
rinsed  with  the comparison water  and  the test person ate several
bites of dry white bread  to "neutralize"  the  taste.  Geometric  mean
detection level values for both tests provided threshold levels of
0.3 ug/1 for taste and 40 yg/1 for  odor  for  2,4-DCP.
     None of  these three studies,  however,  indicated whether the
determined threshold levels made  the water  undesirable  or unfit for
consumption.
                               C-30

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                      CRITERION FORMULATION



Existing Guidelines and Standards



     Presently, no standard for exposure to 2,4-OCP in drinking or



ambient water  has  been set, although  a  standard of  0.1  mg/1 for



2,4-D, a related compound,  has  been set  fNational Academy of Sci-



ences  (WAS), 1977].



Current Levels o€ Exposure



     Human exposure to 2,4-DCP has not been monitored, but a worst



case estimate of 40 yg 2,4-DCP/kg  body weight/day of  exposure was



presented in the Ingestion from Food section.



Special Groups at Risk



     The only  group  expected  to be at risk  from high exposure to



2,4-DCP  is  industrial workers  involved  in  the  manufacturing  or



handling of 2,4-DCP and 2,4-0.  No data were found to relate expo-



sure or body burden to conditions of contact with 2,4-DCP.



Basis and Derivation of Criterion



     Insufficient data exist to indicate  that 2,4-DOP  is a carcino-



genic agent.  The  only study  performed (Boutwell and  Bosch, 1959)



was designed  to  detect promoting  activity,  and the effect of 2,4-



DCP as  a  primary  carcinogen  could not be  evaluated.   Also,  the



route of administration (dermal)  in this  study is inappropriate for



use in the linear model for carcinogenic risk assessment  (see Car-



cinogenicity section).



     Minimal  health  effects data  exist  on the  acute and chronic



effects of  2,4-DCP. Only  one  study of  a chronic  nature (Kobayashi,



et  al.  1972)  was  found.    Kobayashi  and  colleagues  determined  a



chronic (6-month) no-effect level  for  2,4-DCP to be 1,000 ug/q of



diet for mice, which  was  equivalent to 100 mg/kq body weight/dav.





                               C-31

-------
     An equivalent daily dose for a 70 kg adult human would be 7,000

mg/day (100 mg/kg/day x 70 kg).  Applying an uncertainty  factor of

1,000 as suggested by the National Academy of Sciences ^afe Drink-

ing Water Committee (1977),  the Acceptable Daily Intake  (ADI) for  a

70 kg adult  human would be  7,000 mg T  1/000  =  7 mg.  Solving the

equation (2L)(C) + (C) (BCF) (fish consumption/day) = ADI,  the water

quality criterion  (C) can be computed:

           (2L) (C) +  (C)  (40.7) (0.0065) = 7 mg

           (2L)(C) •+•  (C)(0.26455) = 7 mg

          C =   7 "ig
              2.26455L

          C = 3.09 rag/1

where:


     7 mg = the calculated  daily  exposure for a 70 kq person
             (ADI) based on the  above conditions

     2L = amount of drinking water consumed/day

  0.0065 kg = amount of fish consumed/day

     C = maximum  permissible  level  in  water based  on above
         conditions


Thus, a  criterion  level based  entirely  upon  toxicological data

would be 3.09 mg/1.

     Human health is a subjective measurement  in  many  respects.

The  organoleptic properties  of  2,4-DCP  could  conceivably alter

human health by  causing  a  decrease  in water  consumption.   This

might be of particular importance  to  individuals with  certain renal

diseases or  in instances where dehydration  occurs  as a  result of

vigorous exercise, manual labor, or hot weather.
                               C-32

-------
     Since  the  odor and  taste  detection threshold concentrations



for 2,4-dichlorophenol are well below any toxicity-based criterion



level that  may  be  derived,  the  ambient water quality criterion  is



based on organoleptic data.   It  should  be emphasized that this cri-



terion  is based on aesthetic qualities rather than health effects.



However, to  the effect  that  this  criterion  is below the level de-



rived from the chronic toxicity  study of  Kobayashi,  et al.   (1972),



it is likely to also be protective of human health.



     The data of Hoak (1957), Burttschell,  et al.  (1959), and Dietz



and Traud (1978) all  indicated that low microgram concentrations  of



2,4-dichlorophenol in water  are capable  of producing a discernable



odor.   Dietz and Traud  further  observed  a  distinct flavor  altera-



tion of water at sub-microgram levels of  2,4-DCP.  ^he Burttschell,



et al.   (1959) and  Dietz  and  Traud  (1978)  studies did not indicate a



range of responses; however, because  of the  variability of respons-



es inherent  in such  procedures, it  is  certainly possible  that the



odor threshold  for  some  evaluators  (at  least  in  the Burttschell



group)   would extend  downward toward  the  0.65 ug/1  figure  of Hoak.



Thus, the data  from  these three studies  are considered to  be rea-



sonably  mutually   supportive (i.e.,  Hoak's   0.65  ug/1 for odor,



Burttschell's 2.0  ug/1 geometric mean value for  odor,  and Dietz and



Traud's geometric mean values of 40 ug/1 for  odor and 0.3 ug/1 for



taste) .



     Therefore, based on the prevention of undesirable organoleptic



qualities,  the criterion  level  for 2,4-dichlorophenol  in  water  is



0.3 ug/1.  This level should be low enough to prevent detection  of



objectionable organoleptic  characteristics and  far  below  minimal
                               C-33

-------
no-effect concentrations determined in laboratory animals.  As more

substantive and  reliable data become  available  in  the  future,  a

criterion level  based  on human health  effects  mav be more confi-

dently postulated.

     It should be emphasized that data  are needed  in the following

areas to properly evaluate any hazard from 2,4-DCP:


     1)   Monitoring of  worker exposure to 2,4-DCP  in indus-
          tries manufacturing or  using  the chemical.

     2)   Monitoring of  public water  supplies and  industrial
          and  municipal  effluents  to  determine  an expected
          range  of concentrations  under  differing environ-
          mental conditions.

     3)   More definitive studies of residue kinetics of 2,4-
          DCP  in  food  animals which  are  exposed to products
          capable of generating 2,4-DCP.

     4)   Evaluation of  chronic  toxicity,  mutagenicity,  and
          teratogenicity of  2,4-DCP  using  currently accept-
          able techniques.

     5)   A carcinogenicity  study  of  2,4-DCP  using the oral
          route and evaluated according to current  protocols.
                               C-34

-------
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