vvEPA
United States
Environmental Protection
Agency
Office of Water
Regulations and Standards
Criteria and Standards Division
Washington DC 20460
EPA 440/5-80-053
October 1980
Ambient
Water Quality
Criteria for
Hexachlorobutadiene
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AMBIENT WATER QUALITY CRITERIA FOR
HEXACHIOROBUTADIENE (HCBD)
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.
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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 (O.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
no.
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ACKNOWLEDGEMENTS
Aquatic Life Toxicology:
William A. Brungs, ERL-Narragansett David J. Hansen, ERL-Gulf Breeze
U.S. Environmental Protection Agency U.S. Environmental Protection Agency
Mammalian Toxicology and Human Health Effects:
Robert D. Lingg (author) HERL-Cin Roy E. Albert, CAG*
U.S. Environmental Protection Agency u-s- Environmental Protection Agency
Michael L. Dourson (doc. mgr.) ECAO-Cin |Ja"^ JJ?ver,
U.S. Environmental Protection Agency Mob11 Ol1 Corporation
Bonnie Smith (doc. mgr.) ECAO-Cin John L. Laseter
U.S. Environmental Proteciton Agency University of New Orleans
Si Duk Lee, ECAO-Cin Shane Que Hee
U.S. Environmental Protection Agency University of Cincinnati
Joseph Santodonato
Syracuse Research Corporation
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. Oones, B.J. Bordicks,
B.J. Quesnell, C. Russom, R. Rubinstein.
*CAG Participating Members: Elizabeth L. Anderson, Larry Anderson, Ralph Arnicar,
Steven Bayard, David L. Bayliss, Chao W. Chen, John R. Fowle III, Bernard Haberman,
Charalingayya Hiremath, Chang S. Lao, Robert McGaughy, Jeffrey Rosenblatt,
Dharm V. Singh, and Todd W. Thorslund.
iv
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TABLE OF CONTENTS
Criteria Summary
Introduction A-l
Aquatic Life Toxicology 8-1
Introduction B-l
Effects B-l
Acute Toxicity B-l
Chronic Toxicity B-2
Plant Effects B-2
Residues B-2
Miscellaneous B-2
Summary B-2
Criteria B-3
References B-8
Mammalian Toxicology and Human Health Effects C-l
Exposure C-l
Ingestion from Water C-l
Ingestion from Food C-2
Inhalation C-4
Pharmacokinetics C-4
Effects C-5
Acute, Subacute and Chronic Toxicity C-5
Synergism and/or Antagonism C-l7
Teratogenicity C-l 7
Mutagenicity C-l 9
Carcinogenicity C-20
Criterion Formulation C-24
Existing Guidelines and Standards C-24
Current Levels of Exposure C-24
Special Groups at Risk C-24
Basis and Derivation of Criteria C-24
References C-28
Appendix C-35
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CRITERIA DOCUMENT
HEXACHLOROBUTAOIENE
CRITERIA
Aquatic Life
The available data for hexachlorobutadiene indicate that acute and
chronic toxicity to freshwater aquatic life occur at concentrations as low
as 90 and 9.3 gg/1, respectively, and would occur at lower concentrations
among species that are more sensitive than those tested.
The available data for hexachlorobutadiene indicate that acute toxicity
to saltwater aquatic life occurs at concentrations as low as 32 u9/l and
would occur at lower concentrations among species that are more sensitive
than those tested. No data are available concerning the chronic toxicity of
hexachlorobutadiene to sensitive saltwater aquatic life.
Human Health
For the maximum protection of human health from the potential
carcinogenic effects due to exposure of hexachlorobutadiene through
ingestion of contaminated water and contaminated aquatic organisms, the
ambient water concentrations should be zero based on the non-threshold
assumption for this chemical. However, zero level may not be attainable at
the present time. Therefore, the levels which may result in incremental
increase of cancer risk over the lifetime are estimated at 10 , 10 ,
and 10 . The corresponding recommended criteria are 4.47 u9/l> 0.45
ug/l, and 0.045 ug/1, respectively. If the above estimates are made for
consumption of aquatic organisms only, excluding consumption of water, the
levels are 500 ug/l, 50 ug/1, and 5.0 wg/1, respectively.
VI
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INTRODUCTION
Hexachlorobutadiene (HCBO; C^Clg) is produced deliberately in the
United States as a by-product of the manufacture of chlorinated hydrocarbons
such as tetrachloroethylene, trichloroethylene, and carbon tetrachloride.
Secondary production estimates range from 7.3 to 14.5 million pounds (~
5,000 MT) per year (U.S. EPA, 1975a). In 1974, approximately 0.5 million
pounds (- 230 MT) were imported into the U.S. (U.S. EPA, 1975a).
HCBD is used as a solvent for many organic substances; its relatively
low vapor pressure gives it a distinct advantage over some other chlorohy-
drocarbons for this purpose. The largest domestic users of HCBO are chlor-
ine producers, who use it to recover chlorine from "snift" gas which is
cleaned by passage through HCBD. Other applications of HC8D include its use
as an intermediate in the manufacture of rubber compounds and lubricants and
as a fluid for gyroscopes (U.S. EPA, 1975a).
HCBD, a colorless liquid with a faint turpentine-like odor, has a water
solubility of 5 wg/ml at 20°C. It has a melting point of about -21*C, a
vapor pressure of 22 mm Hg at 100°C, and a specific gravity of 1.675 (Haw-
ley, 1977).
Unlike most short chain halogenated aliphatics, hexachlorobutadiene has
a low vapor pressure and, thus, may not volatilize rapidly from the aqueous
environment to the atmosphere. Hexachlorobutadiene has been reported to be
present in domestic drinking water supplies in low concentrations (U.S. EPA,
1975b) and has been detected at concentrations of 1.9 and 4.7 wg/l in water
at two areas near Geismar, Louisiana. These concentrations indicate that
HCBD may be quite persistent in natural waters. However, hexachloroethane,
which is structurally somewhat similar to HCBD and exhibits a vapor pressure
A-l
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-of 0.4 torr at 20*C (Verschueren, 1977) in comparison to 0.15 torr for HCBD
(Pearson and McConnell, 1975), appears to be volatilized rather rapidly from
water. Oil ling (1977) determined a volatilization half-life in water of
40.7 minutes for hexachloroethane initially present at 0.72 mg/1 in an open
system stirred constantly at 200 rpm. Although no specific rate data were
found for HCBD, volatilization may be an important transport process for
this compound in aqueous systems.
Sorption may also be an important process for HC80. The currently re-
viewed literature contains an appreciable amount of information pertaining
specifically to the adsorption of HCBD onto sediments. In a study of the
Mississippi Delta region it was found that the level of hexachlorobutadiene
in water was less than 2 u9/l while the concentration of hexachlorobutadiene
in mud or soil samples exceeded 200 ug/1 (U.S. EPA, 1976b). In this same
study, water samples from the waste of an industrial company in Geismar,
Louisiana, contained from <0.1 ug/l to 4.5 ug/1 HCBD. Levels of HCBD in the
mud, however, reached a maximum of 2,370 ug/1, indicating selective concen-
tration of several orders of magnitude. Leeuwangh, et al. (1975) found that
the concentration of HCBD in uncontaminated sediment after equilibration
with water that contained HCBD was 100 times that found in the water.
Samples taken from Liverpool Bay (England) showed the presence of HCBD,
but rarely at levels greater than 1 ug/l (Pearson and McConnell, 1975).
McConnell, et al. (1975) noted that coarse gravels have little adsorptive
capacity for chlorinated aliphatics, whereas sediments rich in organic
detritus have a much higher adsorptive capacity. The calculated log P
(octanol/water partition coefficient) of 3.74 implies that hexachlorobuta-
diene should be strongly adsorbed by humus material (U.S. EPA, 1979).
A-2
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Environmental contamination by HCBD results primarily during the dis-
posal of wastes containing HCBD from chlorinated hydrocarbon industries
(U.S. EPA, 1976a). Disposal methods include landfill, high temperature
incineration, deep-well injection, and lagoon storage (U.S. EPA, 1975a).
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REFERENCES
Oilling, w.L. 1977. Interphase transfer processes. II. Evaporation rates
of chloromethanes, ethanes, ethylenes, propanes, and propylenes from dilute
aqueous solutions. Comparisons with theoretical predictions. Environ. Sci.
Techno!. 11: 405.
Hawley, G.G. (ed.) 1977. The Condensed Chemical Dictionary. Van Nostrand
Reinhold Co., New York.
Leeuwangh, P., et al. 1975. Toxicity of Hexachlorobutadiene in Aquatic
Organisms. Sublethal Effects of Toxic Chemicals on Aquatic Animals. Ij:
Proc. of Swedish-Netherlands Symp., Sept. 2-5. Elsevier Scientific Publish-
ing Co., Inc., New York. (Abst.)
McConnell, G., et al. 1975. Chlorinated hydrocarbons and the environment.
Endeavor. 34: 13.
Pearson, C.R. and G. McConnell. 1975. Chlorinated C, and C~ hydrocar-
bons in the marine environment. Proc. Royal Soc. Lond. B. 189: 305.
U.S. EPA. 1975a. Survey of industrial processing data. Task I - Hexa-
chlorobenzene and hexachlorobutadiene pollution from chlorocarbon process-
ing. Midwest Res. Inst., Kansas City, Missouri. EPA 560/3-75-003. Off.
Toxic Subst., U.S. Environ. Prot. Agency, Washington, O.C.
A-4
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U.S. EPA. 1975b. Preliminary assessment of suspected carcinogens in drink-
ing water. EPA 560/4-75-003. Off. Toxic Subst., U.S. Environ. Prot. Agen-
cy, Washington, O.C.
U.S. EPA. 1976a. Sampling and analysis of selected toxic substances. Task
IB - Hexachlorobutadiene. EPA 560/6-76-015. Off. Toxic Subst., U.S. Envi-
ron. Prot. Agency, Washington, D.C.
U.S. EPA. 1976b. An ecological study of hexachlorobutadiene. EPA 560/6-
76-010. Off. Toxic Subst., U.S. Environ. Prot. Agency, Washington, D.C.
U.S. EPA. 1979. Water-related environmental fate of 129 priority pollut-
ants. EPA 68-01-3852. U.S. Environ. Prot. Agency, Washington, D.C.
Verschueren, K. 1977. Handbook of Environmental Data on Organic Chemicals.
Van Nostrand Reinhold, New York.
A-5
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Aquatic Life Toxicology*
INTRODUCTION
Available data for hexachlorobutadiene indicate that it is acutely tox-
ic to freshwater aquatic life in the range of 90 to 326 ug/L A physiolog-
ical change in fish blood occurred at much lower concentrations and a chron-
ic effect was observed on fathead minnows at 13.3 yg/1.
Static tests have been conducted with four saltwater species and the
range of 96-hour LC5Q values of 59 to 557 ug/1 is similar to that for
freshwater species.
EFFECTS
Acute Toxicity
A freshwater snail has been exposed to hexachlorobutadiene and the 96-
hour LC5Q value is 210 Hg/l (Table 1).
Goldfish have been tested (Leeuwangh, et al. 1975) and the 96-hour
LC50 is 90 yg/1 (Table 1). The LC50 values for the fathead minnow,
rainbow trout, and bluegill are 102, 320, and 326 ng/1, respectively. All
the tested species, both fish and invertebrate, demonstrated a relatively
narrow range of sensitivity.
Static tests have been conducted with the saltwater mysid shrimp, grass
shrimp, pinfish, and sheepshead minnow; the 96-hour LCc0 values are 59,
32, 399, and 557 ug/l, respectively (Table 1). The fish are approximately
10 times more resistant than the invertebrate species. Both fish species
were affected, but not killed, by hexachlorobutadiene at concentrations be-
low the LCjQ: Cyprinodon variegatus at 240 pg/1 and above, and Lagodon
*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 un-
derstand 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 tox-
icity as described in the Guidelines.
8-1
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rhomboides at 180 yg/1 and above. Within 24 hours of exposure, some swam in
spirals at the surface of the water, while others swam on their sides or lay
motionless, except for opercular movement, on the bottom.
Chronic Toxicity
An embryo-larval test with the fathead minnow (U.S. EPA, 1980) resulted
in a chronic value of 9.3 vg/1 (Table 2). After division of this into the
96-hour LC5g of 102 gg/1, an acute-chronic ratio of 11 is derived.
Plant Effects
No freshwater algal or vascular species have been tested.
Residues
Bioconcentration factors have been determined for goldfish (leeuwangh,
et al. 1975) and range from 920 to 2,300 (Table 3). Laseter, et al. (1976)
obtained bioconcentration factors for an algal species, a crayfish species,
and largemouth bass of 160, 60, and 29, respectively.
Miscellaneous
Laska, et al. (1978) exposed largemouth bass to concentrations of hexa-
chlorobutadiene between 3.43 and 31.95 yg/1 for 10 days and observed ele-
vated blood corticosteroid levels (Table 4).
Summary
There is a narrow range of LC5Q values, for freshwater fish and in-
vertebrate species and hexachlorobutadiene, from 90 to 326 ug/1. The chron-
ic value for the fathead minnow is 9.3 pg/1 with an acute-chronic ratio of
11. There was a wide range of bioconcentration factors of 29 to 2,300 for a
variety of organisms.
As with the freshwater organisms, there is a narrow range of LCcn
values for saltwater fish and invertebrate species of 59 to 5578 ug/1 with
the invertebrate species being more sensitive. No data are available to
estimate chronic toxicity.
B-2
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CRITERIA
The available data for hexachlorobutadiene indicate that acute and
chronic toxicity to freshwater aquatic life occur at concentrations as low
as 90 and 9.3 wg/1, respectively, and would occur at lower concentrations
among species that are more sensitive than those tested.
The available data for hexachlorobutadiene indicate that acute toxicity
to saltwater acuatic life occurs at concentrations as low as 32 ug/1 and
would occur at lower concentrations among species that are more sensitive
than those tested. No data are available concerning the chronic toxicity of
hexachlorobutadiene to sensitive saltwater aquatic life.
B-3
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Table I. Acute values for h*xachlorobutadlene
Species
Snail,
Lyonaea stagnalls
Rainbow trout.
Sal mo galrdnerl
Goldfish.
Car ass 1 us auratus
Fathead minnow,
Plmephales promelas
Blueglll,
Lepomls macrochlrus
Mysld shrimp,
Mysldopsls bah la
Grass shrimp.
Pa 1 aemonetes puqlo
Plnflsh,
Lagodon rhonboldes
Sheep shead minnow,
Cyprlnodon varlegatus
LC50/EC50 Species Acute
Method* lua/n Value (jig/I) Reference
FRESHWATER SPECIES
ft, M 210 210 Leeuwangh. et al. 1975
FT, M 320 320 U.S. EPA, )980a
R, M 90 90 Loauwangh, et al. 1975
FT, M 102 102 U.S. EPA, 1980a
FT. M 326 326 U.S. EPA, 1960a
SALTWATER SPECIES
S, U 59 59 U.S. EPA, 1980b
S, U 32 32 U.S. EPA, 19BOb
S, U 399 399 U.S. EPA, 1980b
S, U 557 557 U.S. EPA, 1980b
* S * static, FT = flow-through, R = renewal, U = unmeasured, M =• measured
No Final Acute Values are calculable since the minimum data base requirements are not met.
B-4
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Table 2. Chronic values for hexachlorobutadlene (U.S. EPA, I980a)
Chronic
Limits Value
Test* (M9/D
FRESHWATER SPECIES
Fathead minnow,
Plmephales promelas
ELS
6.5-13.3 9.3
* ELS = Early life state
Acute-Chronic Ratio
Acute
Value
Fathead minnow,
Plmephales promelas
102
Chronic
Value
(ug/1) Ratio
9.3
11
B-5
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Alga,
OedogonIurn cardlacuro
Crayfish,
Procambarus dark I
Goldfish,
Carnsslus auratus
Largenouth bass,
Mlcropterus salmoIdes
Fable 3. Residues for hexachlorobutadlen*
TIssue
B Ioconcentrat Ion
Factor
FRESHWATER SPECIES
whole body 160
whole body
who Ie body
60
920-2,300
29
Duration
(days)
10
49
10
Reference
Laseter, et a I. 1976
Laseter. et al. 1976
Leeuwangh, et al. 1975
Laseter, et al. 1976
B-6
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Tabla 4. Other data for bexacttlorobutadlene (Laska, at al. 1978)
Result
Species Duration Effect
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REFERENCES
Laseter, J.L., et al. 1976. An ecological study of hexachlorobutadiene
(HCBD). U.S. Environ. Prot. Agency, EPA-560/6-76-010.
Laska, A.L., et al. 1978. Acute and chronic effects of hexachlorobenzene
and hexachlorobutadiene in Red Swamp Crayfish (Procambarus clarki) and se-
lected fish species. Toxicol. Appl. Pharmacol. 43: 1.
Leeuwangh, P., et al. 1975. Toxicity of hexachlorobutadiene in aquatic or-
ganisms. In; Sublethal effects of toxic chemicals on aquatic animals.
Proc. Swedih-Netherlands Symp., Sept. 2-5. Elsevier Scientific Publ. Co.,
Inc., New York.
U.S. EPA. 1980a. Unpublished laboratory data. Environmental Research
Laboratory - Duluth, Minnesota.
U.S. EPA. 1980b. Unpublished laboratory data. Environmental Research
Laboratory - Gulf Breeze, Florida.
B-8
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Mammalian Toxicology and Human Health Effects
EXPOSURE
Ingestion from Water
In 1974, the U.S. Environmental Protection Agency (EPA) con-
ducted a survey of finished water from three New Orleans area water
plants (Keith, et al. 1976). Immediately upstream on the Missis-
sippi River from New Orleans are numerous petrochemical and chem-
ical plants including some producing chlorohydrocarbon compounds.
The concentrations of HCBD found ranged from 0.07 to 0.7 ug/1 HCBD
for the three test sites.
In 1975, a follow-up study conducted by EPA included 10 cities
(U.S. EPA, 1975). HCBD was identified in one of the drinking water
supplies surveyed in this group and the concentration was less than
0.01 ug/1. HCBD has been found in water samples taken from inland
sites bordering the lower Mississippi River (Laseter, et al. 1976).
In this study, a landfill pond near an industrial source was found
to contain 4.49 ug/1 and the corresponding value for the surround-
ing mud was 920 ug/kg (corrected to dry weight). Effluents from
industrial plants suspected of being sources of HCBD contained con-
centrations ranging from 0.04 to 240 ug/1 (Li, 1976).
The conclusions regarding HCBD contamination of drinking water
supplies are: (1) HCBD contamination of U.S. finished drinking
water supplies does not appear to be a widespread problem; (2) the
problem is localized in areas with raw water sources near indus-
trial plants producing HCBD; (3) the physical and chemical
characteristics of HCBD favor rapid extraction of HCBD from
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contaminated water into the surrounding sediment and mud (Laseter,
et al. 1976).
Ingestion from Food
Since the water, soil, and air surrounding certain chloro-
hydrocarbon plants have been shown to be contaminated with HCBD
(Li, et al. 1976), food produced in the vicinity of these plants is
most likely to contain residual levels of HCBD. A survey of food-
stuffs within a 25-mile radius of tetrachloroethylene and tri-
chloroethylene plants was made. Milk, eggs, and vegetable samples
did not contain measurable levels of HCBD. Freshwater fish caught
in the lower Mississippi river contained HCBD residues in the range
of 0.01 to 4.65 mg/kg {Yip, 1976; Yurawecz, et al. 1976).
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 in-
gestion of a lipid-soluble chemical can be estimated from the per
capita consumption of fish and shellfish, the weighted average per-
cent 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 per capita 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
C-2
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the same species to estimate that the weighted average percent
lipids for consumed freshwater and estuarine fish and shellfish is
3.0 percent.
Some bioconcentration factors are available for hexachloro-
butadiene (Laseter, et al. 1976; Leeuwangh, et al. 1975), but the
necessary data concerning percent lipids are not. The equation
"Log BCF = (0.85 Log P) - 0.70" can be used (Veith, et al. 1979) to
estimate the BCF for aquatic organisms that contain about 7.6 per-
cent lipids (Veith, 1980) from the octanol/water partition coeffi-
cient (P). Based on a measured log P value of 1.82 (Dec, et al.
Manuscript), the steady-state bioconcentration factor for hexa-
chlorobutadiene is estimated to be 7.03. 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 per-
cent lipids that is the weighted average for consumed fish and
shellfish. Thus, the weighted average bioconcentration factor for
hexachlorobutadiene and the edible portion of all freshwater and
estuarine aquatic organisms consumed by Americans is calculated to
be 7.03 x 0.395 » 2.78.
Studies on HCBD contamination of food supplies have been re-
ported from several foreign countries. In England, McConnell, et
al. (1975) detected HCBD in fresh milk (0.08 ug/kg), imported
grapes (3.7 ug/kg), and in tomatoes (0.8 jug/kg) grown on a re-
claimed lagoon which was once used as a disposal area for a chemi-
cal plant. In Germany, Kotzias, et al. (1975) found 4 ug/kg HCBD in
evaporated milk, 42 ug/kg in egg yolk, 33 ug/kg in vegetable oil
C-3
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margarine, and 39 and 2 ug/kg in chicken grain feed and laying ra-
tions, respectively.
Inhalation
The air in the vicinity of plants producing tetrachloro-
ethylene, trichloroethylene, and carbon tetrachloride has been
shown to be contaminated with HCBD (Li, et al. 1976). A value as
high as 460 ug/m has been detected in one instance, but generally,
the levels of HCBD detected in air surrounding these chlorohydro-
carbon plants were less than 5 ug/m . There are uncertainties con-
cerning the atmospheric fate of HCBD, but McConnell, et al. (1975)
reported efficient tropospheric destruction of aliphatic organo-
chlorine compounds via photo-oxidation. Unless someone lives or
works in the vicinity of a chemical plant producing HCBD as a major
byproduct, exposure to HCBD through inhalation would not seem to
pose a problem.
PHARMACOKINETICS
Jacobs, et al. (1974) gave oral doses of a mixture of seven
chlorinated hydrocarbons (2 mg/kg/component and 4 mg/kg/component)
to rats daily for up to 12 weeks. Rats sacrificed at 4, 8, and 12
weeks had roughly 7 mg/kg or less HCBD accumulated in fatty tissue
taken from the inner genital and kidney regions. Concentrations of
HCBD on tissues were apparently the same at both dose levels. The
quantity found in the liver, heart, kidney, and blood was less than
that found in the fatty tissue. These results showed that HCBD did
not have a strong tendency to accumulate in fatty tissue of select-
ed organs if administered in a mixture with other chlorinated
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hydrocarbons, some of which were aromatic and accumulated signifi-
cantly in the fat.
EFFECTS
In 1971, Gehring and MacDougall (1971) completed a review of
the toxic properties of HCBD. These authors concluded that the
data available at that time did not allow for a critical evaluation
of any potential hazard associated with long-term, repeated low
level exposure to HCBD. Since then, several excellent studies have
been published on the effects of repeated low level exposure to
HCBD in rats. Kociba, et al. (1977) conducted a two-year study
during which rats were given doses of 0.2, 2.0, and 20 mg/kg/day
HCBD in their diet, and found HCBD induced renal neoplasms at the
highest dose level. Schwetz, et al. (1977), working in the same
laboratory, found that these same dosage levels had virtually no
effect on the reproduction and offspring of rats. Only a slight
decrease in body weight of weanlings at 21 days of age was observed
at the highest dose level.
Acute, Subacute, and Chronic Toxicity
A table summarizing the acute toxicity data on HCBD which was
published prior to 1971 was prepared by Gehring and MacDougall
(1971) in their review and is reproduced in this document as Table
1. Pre-1971 acute toxicity data not included in their table and
acute data published since are summarized in Table 2. Oral LD50
values range from 64 mg/kg in the female weanling rat to 580 mg/kg
in the adult male rat, indicating that HCBD has a relatively high
acute toxicity by the oral route. Toxicity varied with sex and
age differentiation exists. Data on acute dermal and inhalation
C-5
-------
Table 1
Acute Toxic I ty of Hexacltlorobutadiene
Deference
1
2
3
2
2
1
4
3
3
3
3
3
3
3
5
5
5
5
3
5
5
3
3
5
Route Species
Oral Rat
G. Pig
Mice
Dermal G. Pig
Rabbits
Inliala- Rats
tion
G.Pig
Cats
tlo. of
Animals Solvent
6 Corn oil, 3%
4 Propylene glycol
2
2
2
3
3
3
6
5
6
6
6
2
2
4
2
2
Dose
500 mg
126 mg/kg
126 Kg/kg
126 Mg/kg
63 mg/kg
500 ppm
200 ppm
200 ppw
34 ppm
161 ppm
274 ppm
314 ppm
133 ppm
34 ppm
161 ppm
27.5 ppm
34 ppm
161 ppm
Exposure LDSO
Tine mg/Kg
270
350
200
90
87
116
24 hr
7 hr
4 hr
24 hr
4 hr
4 hr
2 hr
3.5 hr
0.83 hr
3.5 hr
0.88 hr
7 hr
3.5 hr
0.88 hr
7 hr
3.5 hr
0.88 hr
Observations
Some Deaths
All died
50t mortality
No death*
No deaths
All died
Two death
One death
No deaths
No deaths
All died
One death
All died
One death
One death
All died
All died
All died
Source: Gehiing t MacDougall, 1971
nlank means no data available.
1 Gulko, et al. 1964.
?. Murzakaev, 1963.
3 Unpublished data, The Dow Chemical Company.
4 Murzakaev, 1966.
5 Treon fc Edwards, 1948.
C-6
-------
Reference
Route
Oral
Table 2
Additional Acute Toxicity Data on Hexachlorobutadiene
Species
Rat(F)
Rat(M)
Rat(P)
Rat(M)
Rat(M)
Hice(M)
IJCDD No. of
Purity Animals Dose
99%
99*
99%
99%
Exposure ^so
Time Kg/Kg
200-400
SftO
65
64
250
80
Observations
21 Jays old
21 days old
Caused hepatic s
disorders, and a
to affect central nervous
of juice
IP
IP
Inhala-
tion
Dermal
Rat(M)
Rat(H)
Rat(H)
Rat(M)
Mice(H)
Mice(M)
Hat
Mice
Guinea Pig
Rabbit
Rabbit
Rabbit
Rat
Rat
529 mg/kg*
16 «g/kg
6800 ppw
6800 pp»*
6800 ppm
5290 «g/kg*
2000 mg/Kg
5000
Single
Single
Converted fro* iil/ky using density of 1.675 mg/iil
Blank Means no data available
Schvetz, et al. 1977.
Gradiski, et al. 1975.
Hazel ton Labs., 1978.
Single
Single
Single
298*
216
105
76
2981
4330
All died in 5 days
No deaths
Lk50 275 win.
Lt50 310 Min.
LtSO 200 ain.
All died
All in three died
Absolute lethal dose
Aerosol dose; estimated concentration
Lt = Lethal time
4 Gradiski, et al. 1974.
5 Chernokan, 1970.
C-7
-------
exposure to HCBD show that HCBD is hazardous when encountered by
these routes.
The subacute and chronic toxicity data are summarized in Table
3. A column has been included in Table 2 and 3 giving the purity of
the HCBD reagent used in each study. This piece of information is
essential in making a final judgment on the usefulness of published
data for assessing the toxicity of a particular substance. Un-
fortunately, no indication of the purity of the reagent has been
given in many of the publications on HCBD toxicity.
A considerable portion of the literature on HCBD toxicity is
reported in the Russian language. For the majority of these publi-
cations, only an abstract was available. All publications on HCBD
toxicity reported in the literature are given in the tables. Those
publications for which translations were available are so labeled
in the references. Because details of the experimental procedures
and results were not included in many of the abstracts of the
Russian articles (and often not in the full translations), only a
limited reference to Russian literature is made in the main body of
this document. None of the Russian articles attributed carcinogen-
ic properties to HCBD.
Renal damage, as evidenced by renal tubular epithelial degen-
eration, necrosis, and an increase in the kidney weight:body weight
ratio occurred in female rats receiving 30, 65, or 100 mg/kg/day
HCBD for 30 days in their diet (Kociba, et al. 1971). No histo-
pathologic changes were observed at the 3 mg/kg/day HCBD dosage
level. Other observed effects were: decreased food consumption and
body weight gains for female rats consuming 10, 30, 65, and 100
C-8
-------
Table 3
Subacute and Chronic Toxicity of Ilexachlorobutadiene
Uotercnce Route Species
1 Oral Rat(M)
(diet) Rat(F)
Rat(H)
Rat(F)
Rat(M)
Rat(F)
2 Oral Rat(F)
(diet) Rat(F)
Rat(F)
Rat(F)
Rat(F)
Rat(F)
3 Oral Rat(M)
(diet) Rat(F)
Rat(M)
Ra t ( F )
Rat(M)
Rat(l-')
No. of
Animals
39
40
40
40
40
40
4
4
4
4
4
4
12
24
10
20
12
24
% HCDD
Purity
99
99
99
99
99
99
99
99
99
99
99
99
99
99
99
99
99
99
Dose
20 rag/ kg/day
20 mg/kg/day
2 lag/kg/day
2 mg/kg/day
0.2 ng/kg/day
0.2 mg/kg/day
1 mg/kg/day
3 mg/kg/day
10 ng/kg/day
30 mg/kg/day
65 mg/kg/day
100 mg/kg/day
0.2 mg/kg/day
0.2 mg/kg/day
2.0 my/kg/day
2.0 mg/kg/day
20 mg/kg/day
20 mo/ kg/day
Duration
22 no
24 mo
22 mo
24 no
22 mo
24 mo
30 days
30 days
30 days
30 days
30 days
30 days
148 days
148 days
148 days
148 days
148 days
148 days
Observations
Renal tubular neoplasms; metastasis to
the lung
Increased urinary coproporphyr in, increase
in renal tubular epithelial hyperplasia
Uf-t o f f &r* i- a
nU CL I CCL 3
No effects
Marginal change in kidney/body weight ratio;
no pathologic alterations
Decreased body weight gain; no pathologic
alterations
Renal tubular epithelial degeneration,
individual cell necrosis and regeneration,
decreased body weight gain; increase in
mean kidney: body weight ratio; increase
in hemoglobin concentration
No effects among adults or neonates
No effects among adults or neonates
Kidney "roughened,* mottled cortex; other
kidney changes which normally occur appeared
to be accentuated
Accentuation of normal kidney changes;
one had renal lesions identical to those on
20 mg/kg/day; no effects on neonates
Change in kidney body weight ratio; kidney
roughened with mottled cortex; renal tubular
dilation and hypertrophy with foci ot renal
tubular epithelial degeneration and lecjurie ra-
tion
Renal tubular dilation and hyper tiophy
with foci of renal tubular epithelial do-
generation and regeneration; decreased
value of body weight and heart, increased
values for relative weight of brain and
kidney; slight decrease in body weight
of neonates at time of weaning.
C-9
-------
Table 3 (Continued)
Heference
Route
Oral
Oral
Oral
Spec lea
Rat
Rat
Rat
Rat
Guinea
P19
No. of % 11CDD
Animals Purity Dose
0.0005 Ma/kg/day
0.004 Mg/kg/day
0.02 Mg/ kg/day
8.4 mg/kg
100 Kg/kg
60 Kg/kg
90 Mg/kg
Duration
6 MO
4 MO
2 MO
Single
Nas
Weak
High
Seve
chan
Deco
days
Observations
7,8
10
11
Oral
Ural
Parent-
eral
Dog
Dog
Guinea
Pig
Rat(F)
Rat(F)
Rat(F)
IP
M i cc-
20
1 mg/kg/day
O.OS ng/kg/day
0.004-2 Mg/kg/day
0.004 Mg/kg
0.04 Mg/kg
7.0 mg/kg
4 *g/kg
8 mg/kg
Nas at threshold level with respect to
6 MO
45 days
7 MO
6 MOS
6 MOS
6 BOS
*
wk to
total of
52 mg
wk to
total of
96 Mg
Severe necrotic nephrosis, as well as abnoiwal
changes in the brain, liver, and other
internal organs
DecoMpensatory acidosis Most significant at 5
daysi those surviving 15 days, the blood
indicators of acid-base equilibrium wore
normalized
Administered to puppies from birth to 6 mog.
Increased secretion of total N-containing
compounds. Increased vol. and total acidity of
the gastric juice
Administered 1.5 to 3 Months postnatal;
increased total vol., acidity, and amount of
I1C1 and chloride secreted by the stomach.
2 Mg dose caused a decrease in -Sll gioup
cone, in blood plasma without change in
blood protein plasma spectrum
No effect on rate of forming positive condi-
tioned reflexes
Acceleration of differentiation was noted.
Decrease in the Sll content of the cerebral
cortex homogenate; disturbed ability of
the animal to form conditioned reflex con-
nections; Exhibited morphological changes
in the liver, kidneys, and cardiac muscle
in the fora ot parenchyMatous dystrophy
No statistically significant increase in
the production of adenomas
C-10
-------
Table 3 (Continued)
He EC: to nee
'n "
14
15
16
17
10
Route
Subcut-
aneous
Inhala-
tion
Inhala-
tion
Inhala-
tion
Cutan-
eous &
ocular
Dermal
Inhala-
tion
oral, and
topical
No. of
Species Animals
Rat(F)
Rat
Mice
Rat
Rat (4 M) 4
Rat(4 F) 4
Rat(4 M) 4
Rat(4 F) 4
Rat(4 M) 4
Rat (4 F) 4
Rat
Rat
Rat, Mice,
Guinea pig
& cats
t I1CBD
Purity Dose
20 ng/kg
•>
24 «g/m^-air
24 mg/m -air
0.01 mg/« -air
25 ppm
25 ppn
100 ppa
100 ppa
10 or 5 ppn
10 or 5 ppm
1,675 mg/kg*
8.4 rag/kg
10 mg/kg
Duration
Single
7 mos
7 mos
6 mos
(5 hr daily)
15 daily
for 6 hrs
12 daily
for 6 hrs
15 daily
Cor 6 hrs
Single
24 hrs
Single
Single
Observations
Administered to pregnant mice, caused distur-
bances of motor coordination; weight loss,
rymphocytosis, neutropenia, myelcy tosis,
death occurred in 100% of the offspring
within 3 mos. after injection
Caused no alterations
No effects observed
Caused respiratory difficulty; decreased
weight gain and pathologic injury to the
tubular epithelium of the kidneys
Severe toxicity including death
Caused no toxicity except for retarded
weight gain in females at 10 ppra
Irritant
Mild to node rate erythema
Acute and chronic toxicity experiments
performed on large groups of rats, nice.
guinea pigs, and cats. tlematological study
showed leucocy tosis, lymphocytes is and de-
creased erythrocytic permiability to water.
immunological depression and decreased anti-
body formation were observed
Largest concentration of IICltD found in proxi-
mal sections of the nephrons: dystropic
changes in kidneys caused by 8.4 mg/kij;
necrosis at 10 m
-------
Table 3 (Continued)
References for. Table 3
1. Kociba, et al. 1977
2. Kociba, et al. 1971
3. Schwetz, et al. 1977
4. Potetyaeva, 1973
5. Diaitrlenko and Vasilos, 1972
6. Popovich, 1975
7. Boranova, 1974a
8. Boranova, 1974b
9. Kravitskaya and Docanova, 1974
10. Murzakaev, 1965
11. Murzakaev, 1967
12. Theiss, et al. 1977
13. Poteryaeva, 1966
14. Gulko, et al. 1964
15. Poteryaeva, 1972
16. Gage, 1970
17. Dupcat, et al. 1976
18. Numma and Lawless, 1975
19. Poteryaeva, 1971
20. Shroit, et al. 1972
C-12
-------
mg/kg/day HCBD in their diet; depletion of abdominal fat at 65 and
100 mg/kg/day; minimal hepatocellular swelling at 100 mg/kg/day;
and hemoconcentration at 10, 30, 65, and 100 mg/kg.
Dietary ingestion of 20 but not 2.0 or 0.2 mg/kg/day HCBD for
two years caused depression of the body weight gain of both male
and female rats (Kociba, et al. 1977). Evaluation of the mean or-
gan weights and organ:body weight ratios for rats killed at the
termination of this study indicated the males had no alterations in
weights of the brain, heart, and liver relative to control values.
The relative and absolute weights of the kidneys of males ingesting
the 20 mg/kg/day were found to be increased. For females, the body
weight loss was accompanied by a significant decrease in the ab-
solute weight of the heart and liver, and an increase in the rela-
tive weight of the kidney. A decreased survival rate was observed
for males but not for females at 20 mg/kg/day HCBD.
Extensive gross and microscopic pathological examination of
all rats necropsied during the course of this study was conducted.
Significant abnormalities were observed in the urinary systems of
rats receiving the 20 mg/kg/day dosage level. Among effects re-
lated to HCBD treatment was an increase over the controls in renal
tubular neoplasms in both male and female rats. The effects at 2
mg/kg/day HCBD were slight, including possible renal tubular epi-
thelial hyperplasia. No effects were observed in rats receiving
0.2 mg/kg/day HCBD in their diet for two years.
Blood, serum, and urine samples were analyzed for a wide var-
iety of clinical indicators. A statistically significant increase
in urinary coproporphyrin was observed in male rats ingesting 20
C-13
-------
mg/kg/day HCBD for 12 months, in females ingesting 2.0 mg/kg/day
for 14 months, and in females ingesting 20 mg/kg/day HCBD for 24
months.
A study of the effects of HCBD on reproduction in rats in-
cluded gross and microscopic examination of internal organs, bone,
and other tissues (Schwetz, et al. 1977). Pathological changes
considered to be related to ingestion of HCBD were found in the
kidneys of both male and female rats at dietary dose levels of 2 and
20 mg/kg/day HCBD for 148 days. Kidneys from male rats at these two
dose levels were "roughened" and had a mottled cortex. There was
renal tubular dilation and hypertrophy with foci of renal tubular
epithelial degeneration and regeneration in kidneys from male and
female rats on 20 mg/kg/day HCBD. No effects were observed at 0.2
mg/kg/day.
An effect on the central nervous system was observed at a 7
mg/kg (frequency of administration unknown) dose level as a reduc-
tion in the response capability of the rat to conditioned reflexes
(Murzakaev, 1967). Results at 0.04 mg/kg were not statistically
significant, while 0.004 mg/kg HCBD gave no indication of a neuro-
toxic response. Poteryaeva (1973) measured urinary acid-base equi-
librium, serum peroxidase, residual nitrogen, and neuromuscular
chronaxie in rats receiving daily oral doses of HCBD. HCBD tox-
icity was observed at 0.02 mg/kg/day for one month, at 0.004
mg/kg/day for four months, and threshold toxicity at 0.0005
mg/kg/day after six months.
Kidney damage is also induced by inhalation of HCBD. Fifteen
daily six-hour exposures of rats to 25 ppm HCBD caused respiratory
C-14
-------
difficulty, decreased weight gain, and pathologic injury to the
tubular epithelium of the kidneys. Twelve daily six-hour exposures
to 100 ppm HCBD caused more severe toxicity, including the death of
some rats. Fifteen daily six-hour exposures to 5 or 10 ppm HCBD
caused no toxicity except for retarded weight gain in females ex-
posed to 10 ppm (Gage, 1970).
Chernokan (1970) determined the dermal LD50 of HCBD for the
rat at 4,330 mg/kg. Animals receiving 1/5 dermal LD5Q showed gen-
eral weight loss, sluggishness, and paresis of the extremities
within one day of the start of the experiment, and all had died by
the fifth to sixth day. With repeated applications of 1/20 dermal
LDen HCBD, features of intoxication began showing up by the sixth
day which were expressed in weight loss, flaccidity, and rigidity
of muscles. The red blood cell was the most sensitive initial in-
dicator of toxicity as manifested by a loss in hemoglobin level and
a drop in erythrocyte count.
Boranova (1974b) observed that oral administration of 1 mg/day
HCBD to puppies from birth to six months increased excretion of
total nitrogen-containing compounds and of urea nitrogen starting
on the 20th day of growth. Total acidity of gastric juices and con-
tent of free HC1 also were increased. A single dose of 60 or 90
mg/kg to guinea pigs caused decompensatory acidosis which was most
significant at five days (Popovich, 1975). Indicators of acid-base
equilibrium in blood and urine tended toward normalization at 15
days.
The kidney appears to be the organ most sensitive to HCBD.
Possible chronic effects are observed at doses as low as 2 to 3
C-15
-------
ing/kg/day (Kociba, et al. 1971, 1977; Schwetz, et al. 1977). Renal
tubular neoplasms were observed during a two-year study in which 20
mg/kg/day was administered to rats in their diet (Kociba, et al.
1977). Single oral doses as low as 8.4 mg/kg have been observed to
have a deleterious effect on the kidney {Shroit, 1972). Neurotoxic
effects have been reported to occur at dose levels as low as 4
ug/kg/day {Poteryaeva, 1973; Murzakaev, 1967). Acute HCBD intoxi-
cation affects acid-base equilibrium in blood and urine (Popovich,
1975? Poteryaeva, 1971). Some investigators report a cumulative
effect for HCBD during chronic dosing by dermal (Chernokan, 1970)
or oral (Poteryaeva, 1973) routes. An increase in urinary copor-
phyrin was observed in rats receiving 2 mg/kg/day and 20 mg/kg/day
HCBD for up to 24 months (Kociba, et al. 1977).
In Russia, where HCBD is used as a soil fumigant for grape
phylloxera, exposure of vineyard workers has occurred. Measure-
ments showed that HCBD persisted in the air at levels from 0.0012
to 0.01 mg/1 for up to 5 days depending upon weather, application
method, and soil tillage (Krasniuk, et al. 1969). A medical exam-
ination was done of 205 workers, 153 with 4 years exposure to HCBD
and polychlorobutane-80 (a mixture of partly chlorinated 1,3 buta-
dienes) and 52 working under the same conditions but with no expo-
sure to these compounds. Arterial hypotension, as a rule, was
noted in workers exposed to HCBD and polychlorobutane-80. Heart
changes, including myocardial dystrophy, were found more frequently
in workers in contact with the fumigant than in workers with no
contact. Dyspeptic signs and periodic epigastric pains associated
with food intake were observed in 38 subjects. Nervous disorders
C-16
-------
a.nd upper respiratory tract changes occurred more frequently in
exposed than in nonexposed workers. Details and tests for statis-
tical significance were not presented.
McConnell, et al. (1975) reported finding HCBD in human liver
samples at 12 ug/kg wet tissue and in body fat at around 4 pg/kg.
Synergism and/or Antagonism
Murzakaev (1967) found a decreased -SH group content in the
cerebral cortex homogenate and blood serum of rats receiving a 7
mg/kg parenteral dose of HCBD over a six-month span (frequency of
dose not given). Mizyukova, et al. (1973) determined the relation-
ship between the structure and detoxifying power of thiols and
amines against HCBD. Rats poisoned with 300 mg/kg or larger doses
of HCBD were given thiols or amines 20 to 30 minutes before and
immediately after poisoning. The antidotes were administered oral-
ly in twofold molar excess relative to the poison. As determined
from the survival rates, mercaptide, cysteine, and especially uni-
thiol were highly effective antidotes for HCBD. The ethanol-
ammonium salts of 2,3-dimercaptopropane sulfonic acid were even
more powerful antidotes than unithiol, while the ethanolamines
themselves, either alon« or combined with unithiol, were ineffec-
tive. The reduction in free -SH groups following HCBD injection
and the effective antidote with mercapto (-SH) compounds suggests
that SC9D reacts readily with compounds containing these groups.
Teratogenicity
Nonpregnant female rats were given single 20 rag/kg subcutan-
eous doses of HCBD and then bred (Poteryaeva, 1966). The course
Q' the subsequent pregnancy and its outcome were followed. The
C-17
-------
pregnancy rate for the treated rats was the same as in the control
group. No stillbirths occurred, but all the offspring of the rats
that had received HCBD died within 3 months of birth with 38 of 86
dying between days 1 and 28, and the rest of the offspring dying
between days 29 and 90. In the control group, 13 of 61 newborns
died within 14 days of birth with no deaths occurring thereafter
for up to 90 days. The weights of the young rats from the dosed
mothers were markedly lower than the controls. At 2% months, when
the rats showed marked toxic effects, gross pathologic changes were
noted in the internal organs at autopsy. Damage to the kidney was
observed in the form of glomerulonephritis. Blood smears showed
degenerative changes in the red cells,
Schwetz, et al. (1977) studied the dietary effects of HCBD on
reproduction in rats. Male and female rats were fed dose levels of
0.2, 2.0, or 20 mg/kg/day HCBD 90 days prior to mating, 15 days dur-
ing mating, and subsequently throughout gestation and lactation.
The toxic effects observed in these rats are given in Table 3.
Signs of toxicity observed among the adult rats at the two higher
dose levels were decreased weight gain, low food consumption, and
alterations in the kidney structure in the form of a mottled cor-
tex. The only effect on weanlings consisted of a slight decrease
in body weight at 21 days of age at the 20 mg/kg/dose level. All
parameters of the reproductive process including neonatal growth,
survival, and development were normal at levels of 0.2 or 2.0
nig/kg/day.
The discrepancy between the survival rates and observations on
young rats reported by Schwetz, et al. (1977) and Poteryaeva (1966)
C-18
-------
suggests the need for more research on the teratogenic effects of
HCBD. The difference in routes of administration and absorption
may account for differences in survival rates and toxic effects.
Schwetz, et al. (1977) reports a 94 percent survival rate for new-
born controls to day 14, whereas Poteryaeva (1966) reports only a
79 percent survival rate for controls.
A study published by Schwetz, et al. (1974) dealt with the ef-
fects of HCBD on reproduction in Japanese quail (Coturnix
japonica). Adult male and female Japanese quail were fed diets
containing 0.2, 3, 10, or 30 mg/kg of HCBD for 90 days. The birds
showed no evident signs of toxicity during the study. HCBD at all
dose levels had no effect on body weight, demeanor, food consump-
tion, egg production, the fertility and hatchability of eggs, the
survival of hatched chicks, and eggshell thickness. In addition,
at the termination of the study there were no gross or histopatho-
logic changes evident in the organs or tissues of birds that could
be related to treatment.
Mutagenicity
Taylor (1978) has tested the mutagenic potential of HCBD on £>.
typhimurium TA 100. Due to problems in solubilizing HCBD in the
test system, Taylor concluded that HCBD mutagenicity was 'not
proven.' A dose-dependent increase in reversion rate in the ab-
sence of activation was observed, but the usual criterion for muta-
genicity (at least a twofold increase in reversion rate; U.S. EPA,
1977) was not reached. The final conclusion was that HCBD was non-
mutagenic. HCBD was also found to be nonmutagenic in the presence
of an activating system.
C-L9
-------
C-arcinogenicity
Kociba, et al. (1977) completed a two year study of the chron-
ic effects of HCBD in the rat. The results of the study are covered
in Table 4. The most significant finding of these workers was the
development of renal tubular neoplasms in the kidneys of rats re-
ceiving 20 mg/kg/day HCBD in their diet. Histological examination
of the tumors revealed renal tubular adenomas and adenocarcinomas.
Metastasis to the lung was observed in two cases.
The HCBD was 99 percent pure. Test diets to supply 0, 20, 2,
or 0.2 mg/kg/day were prepared weekly for the first three months of
the study and monthly thereafter with concentrations of test mater-
ial being adjusted to maintain the designated dosages on a
^9/kg/day basis. Seven week-old male and female Sprague-Daw ley
(Spartan substrain) specific-pathogen-free-derived rats were used.
The rats were randomized into test groups of 39 to 40/sex/dose
level plus 90/sex for controls.
In a 30-day toxicity study by Kociba, et al. (1971), female
rats received HCBD in doses ranging from 1 to 100 mg/kg/day; no ef-
fects were observed in rats receiving 3 mg/kg/day, while effects
were only marginal in rats receiving 10 mg/kg/day. The kidney was
identified as the target organ. The no-effect-observed levels are
roughly in the same range as those reported by other investigators,
and the kidney has been identified repeatedly as the target organ.
Feeding of the male rats continued for 22 months while the fe-
males were treated for 24 months. During the course of the study,
gross and microscopic examinations were conducted on: (1) represen-
tative portions of all major organs; and (2) any tissue having a
C-20
-------
Table 4
i
Response to HCDD Feeding in Male and Female Rats
Dose
mg/kg/day
20.0
2.0
0.2
Control
Numbers
Male rat
9/39
0/40
0/40
1/90
% Response
Male rat
23
0
0
1.1
Numbers
Female rat
3/40
0/40
0/40
1/90
% Response
Female rat
7.5
0
0
1.1
Observations
Renal tubular adeno-
carcinomas; Undifferen-
tiated carcinoma; metas-
tasis to the lung
Nephroblastoma
Source: Kociba, et al. 1977
C-21
-------
gross lesion suggestive of a significant pathological process taken
from rats killed in a moribund condition. Terminal necropsy exam-
inations were conducted on rats surviving the full term on the test
diets. There was a significant increase in the mortality of the
male rats ingesting 20 mg/kg/day HCBD during the last two months of
the study. This was not the case with the males on the lower dose
levels or in females at any dose level. Fifty percent of the high
dose males had died by 18 months, while the survival rate for fe-
males was 21 to 22 months before a 50 percent death count was ob-
served.
No significant increase in incidence of neoplastic lesions re-
lated to ingestion of HCBD was observed in any organ, tissue, or
skeletal samples other than in the kidney. In the males receiving
20 mg/kg/day of HCBD, 18 percent (9/39) had renal tubular neoplasms
which were classified as adenomas or carcinomas; 7.5 percent (3/40)
of the females on the high dose developed renal carcinomas. Meta-
stasis to the lung was observed in one case for both male and female
rats. A nephroblastoma developed in one of the male control rats
(1/90 or 1.1 percent) and in one of the female controls (1/90 of 1.1
percent). No carcinomas were observed in the kidneys of the con-
trol rats, and no nephroblastomas were observed in the kidneys of
the rats maintained on diets containing HCBD. Table 4 summarizes
the data from Kociba, et al. (1977).
The production of lung adenomas in strain A mice following
multiple intraperitoneal (i.p.) injections of HCBD was investigated
(Theiss, et al. 1977). Doses of 4 mg/kg and 8 mg/kg were admin-
istered i.p. (tricaprylin vehicle) three times a week to groups of
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20 male mice six to eight weeks old. Treatment continued until
totals of 52 mg (4 mg/kg) and 96 mg (8 mg/kg) were administered.
Twenty-four weeks after the first injection, the mice were killed.
There was no statistically significant increase in the mean number
of lung tumors per test mouse as compared to vehicle-treated con-
trols/ nor was a dose-response relation obtained.
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CRITERION FORMULATION
Existing Guidelines and Standards
There are no guidelines or standards of record other than a
standard of 10 jag/m for inhalation referenced by Poteryaeva (1972)
of the Soviet Union.
Current Levels of Exposure
The analytical data available on the distribution of HCBD in
the environment suggest that exposure is a localized problem, po-
tentially affecting those living in areas with nearby chemical
plants producing parent compounds for which HCBD is a by-product.
However, due to the limited number of air and water samples taken,
it is difficult to estimate the level of exposure even to those
populations living in areas where the heaviest exposure might be
encountered.
Special Groups at Risk
A special group at risk would be those workers in an indus-
trial environment where concentrations of HCBD in the air might be
present.
Basis and Derivation of Criteria
HCBD exhibits acute, subacute, and chronic toxicity in animal
test systems; the overall data can be reviewed in Tables 1-4.
The kidney appears to be the organ most sensitive to HCBD.
Chronic effects are observed at doses as low as 2 to 3 mg/kg/day in
rats (Kociba, et al. 1971, 1977; Schwetz, et al. 1977). Renal
tubular neoplasms were observed during a two-year study in which 20
mg/kg/day was administered to rats in their diet. Single oral
doses as low as 8.4 mg/kg have been observed to have a deleterious
C-24
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e.ffect on the kidney (Shroit, 1972). The carcinogenic effects of
renal tubular adenomas and adenocarcinomas were strongly demon-
strated at the 20 mg/kg/day dosage. Even though HCBD was not
determined to be mutagenic in £. typhimurium (Taylor, 1978), since
the mutation rate was not double the background rate, the compound
is a carcinogen, as strongly indicated by the study of Kociba, et
al. (1977).
The evidence of carcinogenicity is sufficient to conclude that
HCBD is a suspect human carcinogen. As carcinogens can conserva-
tively be assumed to have a nonthreshold dose/response character-
istic, the carcinogenic effect is the most significant exposure ef-
fect from which to estimate an ambient water quality criterion
value.
Under the Consent Decree in NRDC v. Train, criteria are to
state "recommended maximum permissible concentrations (including
where appropriate, zero) consistent with the protection of aquatic
organisms, human health, and recreational activities." Hexachloro-
butadiene is suspected of being a human carcinogen. Because there
is no recognized safe concentration for a human carcinogen, the re-
commended concentration of HCBD in water for maximum protection of
human health is zero.
Because attaining a zero concentration level may be infeasible
in some cases and in order to assist the Agency and states in the
possible future development of water quality regulations, the con-
centrations of HCBD corresponding to several incremental lifetime
cancer risk levels have been estimated. A cancer risk level pro-
vides an estimate of the additional incidence of cancer that may be
C-25
-------
expected in an exposed population. A risk of 10 for example,
indicates a probability of one additional case of cancer for every
100,000 people exposed, a risk of 10 indicates one additional
case of cancer for every million people exposed, and so forth.
In the Federal Register (44 FR 15926) notice of availability
of draft ambient water quality criteria, EPA stated that it is con-
sidering setting criteria at an interim target risk level of 10 ,
10" , or 10 as shown in the table below.
Risk Levels .
Exposure Assumptions and Corresponding Criteria
10"7 10_'6 1£~5
2 liters of drinking 0.045 jug/1 0.45 jjg/1 4.47 pg/1
water and consumption
of 6.5 grams of fish,
shellfish (2).
Consumption of fish 5.00 jug/1 50.0 jjg/1 500 ug/1
and shellfish only.
(1) Calculated by applying a linearized multistage model as dis-
cussed in the Human Health Methodology Appendices to the
October 1980 Federal Register notice which announced the
availability of this document. Appropriate bioassay data used
in the calculation are presented in Appendix I. Since the ex-
trapolation model is linear at low doses, the additional life-
time risk is directly proportional to the water concentration.
Therefore, water concentrations corresponding to other risk
levels can be derived by multiplying or dividing one of the
risk levels and corresponding water concentrations shown in
the table by factors such as 10, 100, 1,000, and so forth.
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f2) Approximately 1 percent of the HCBD exposure results from the
consumption of aquatic organisms which exhibit an average
bioconcentration potential of 2.78-fold. The remaining 99
percent of HCBO exposure results from drinking water.
Concentration levels were derived by assuming a lifetime expo-
sure to various amounts of HCBO, (1) occurring from the consumption
of both drinking water and aquatic life grown in water containing
the corresponding HCBD concentrations, and (2) occurring solely
from the consumption of aquatic life grown in the waters containing
the corresponding HCBD concentrations. Because data indicating
other sources of HCBD exposure and their contributions to total
body burden are inadequate for quantitative use, the figures re-
flect the incremental risks associated with the indicated routes
only.
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APPENDIX I
Derivation of Criterion for Hexachlorobutadiene
During the two year feeding study of Kociba (1977) in rats,
renal tubular adenomas and carcinomas were observed in males with
significantly higher incidence in animals fed 20 mg/kg/day than
control animals. Using a fish bioaccumulation factor of 2.78, the
parameters of the extrapolation model are:
Dose Incidence
(mg/kg/day) (no. responding/no, tested)
0.0 1/90
0.2 0/40
2.0 0/40
20.0 9/39
le = 669 days w * 0.610 kg
Le - 730 days R = 2.78
L = 730 days
With these parameters the carcinogenic potency for humans, q, ,
is 0.07752 (mg/kg/day)~ . The result is that the water concentra-
tion should be less than 4.47 micrograms per liter in order to keep
the individual lifetime risk below 10 .
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