> snvironmaruaJ ?*o :ectiofi aliens and S(or..:afcs Octooar '980
,Aa«ocv C.-.t«n« and Standaras Oivision mvjfW*- o„
Wainingron DC 20460 QO»J>
Ambient
Water Quality
Criteria for
Acenaphthene
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AMBIENT WATER QUALITY CRITERIA FOR
ACENAPHTHENE
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
i.
-------
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)(1) 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)(1) 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 conments 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
(0.0".C. 1976), modified, 12 £RC 1333 {o.D.C. 1979).
The term "water quality criteria" is used in two sections of the
Clean Water Act, section 304 (a)(1) and section 303 (c)(2). The tern 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
'seal environmental conditions and human exposure patterns before
corporation into water quality standards. It is not unti1 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
iii
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ACKNOWLEDGEMENTS
Aquatic Life Toxicology:
William A. Srungs, ERL-Narragansett John H. Gentile, ERL-?larragansev.-
U.S. Environmental Protection Agency U.S. Envi ronmental Protection Acer.cv
Mafnnalian Toxicology and Human Health E
Anne Trontell (author)
Energy Resources Company
Steven D. Lutkenhoff (doc. mgr.)
ECAO-Cin
U.S. Environmental Protection Agency
Bonnie Smith (doc. mgr.)
ECAO-Cin
U.S. Environmental Protection Agency
Patrick Dugan
Ohio State University
Rolf Hartung
University of Michigan
Fred Kopfler, HERL
U.S. Environmental Protection Agency
Lei and L. Smith
University of Texas Medical School
Woodhall Stopford
Duke University Medical Center
Debbie Geismar (author)
Energy Resources Company
Mary F. Argus
Tulane University
Julian Andelman
University of Pittsburgh
Patrick Durkin
Syracuse Research Corporation
Betty LaRue Herndon
Midwest Research Institute
Fumio Matsamura
Michigan State University
Jerry F. Stara
ECAO-Ci n
U.S. Environmental Protection Agency
Jonathcn Ward
University of Texas Medical Branch
Technical Support Services Staff: O.J. Reisman, M.A. Garlough, B.L. Zwayer,
P.A. Daunt, K.S. Edwards, T.A. Scanaura, A.T. Pressley, C.A. Cooper,
M.M. Oenessen.
Clerical Staff: C.A. Haynes, S.J. Faehr, L.A. '.lade, D. Jones, 3.J. Bordicks,
B.J. Quesnell, C. Russom, B. Gardiner.
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TABLE OF CONTENTS
Page
Criteria Sunwary
Introduction A-l
Aquatic Life Toxicology B-i
Introduction 8-1
Effects B-l
Acute Toxicity B-l
Chronic Toxicity 8-1
Plant Effects B-2
Residues 6-2
Summary B-2
Criteria B-3
Reference0 8-3
Mamnalian Tc- alogy and Human Health Effects: C-l
E.xposur C-l
Inges :n from Water C-l
Inges :n from Food C-2
Inhalation C-3
Dermal C-3
PharmacoKinetics C-4
Absorption and Distribution C-4
Metabolism C-4
Excretion C-4
Effects C-4
Acute, Subacute, and Chronic Toxicity C-4
Synergism and/or Antagonism C-7
Teratogenicity C-8
Mutagenicity C-3
Other Cellular Effects C-9
Carcinogenicity C-18
Criterion Formulation C-21
Existing Guidelines and Standards C-21
Current Levels of Exposure C-21
Special Groups at Risk C-21
Basis and Derivation of Criterion C-21
References C-23
v
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CRITERIA OOCUMENT
ACENAPHTHENE
CRITERIA
Aouatic Ufa
The available data for acenaohthene indicate that acute toxicity to
freshwater aauatic life occurs at concentrations as low as 1,700 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
acenaohthene to sensitive freshwater aauatic animals but toxicity to fresh-
water algae occur at concentrations as low as 520 ug/1.
The available data for acenaohthene indicate that acute and chronic tox-
icity to saltwater aauatic life occur at concentrations as low as 970 and
710 ug/1, respectively, and would occur at lower concentrations among spe-
cies that are more sensitive than those tested. Toxicity to algae occurs at
concentrations as low as 500 ug/1.
Human Health
Sufficient data are not available for acenaphthene to derive a level
which would protect against the potential toxicity of this compound. Using
available organoleptic data, for controlling undesirable taste and odor
auality of ambient water, the estimated level is 0.02 mg/1. It should be
recognized that organoleptic data, as a basis for establishing a water aual-
ity cr1t«r1*, have limitations and have no demonstrated relationship to po-
tential adverse human health effects.
vi
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INTRODUCTION
Acenaphthene (1,2-dehydro-acenaphtnylene or 1,8-ethylene-naphthalene)
occurs in coal tar produced during the high temperature carbonization or
coking of coal. It is used as a dye intermediate, in the manufacture of
some plastics, as an insecticide and fungicide, and has been detected in
cigarette smoke and gasoline exhaust condensates. Acenaphthene is a poly-
nuclear aromatic hydrocarbon with a molecular weight of 154 and a formula of
C12h10*
The compound is a white crystalline solid at room temperature with a
melting range of 95 to 97*C and a boiling range of 278 to 28Q*C (lidner,
1931). The. vapor pressure is less than 0.02 mm Hg. Acenaphthene is soluble
in water (100 mg/1), but solubility is greater in organic solvents such as
ethanol, toluene, and chloroform.
Acenaphthene will react with molecular oxygen in the presence of alkali-
earth bromides to form acenaphthequinone (Oigurov, et al. 1970). In the
pre -nee of alkali-earth metal hydroxides, acenaphthene reacts with ozone to
produce 1,8-naphthaldehyde carboxylic acid (Menyailo, et al. 1971). Ace-
naphthalene can be oxidized to aromatic alcohols and ketones using transi-
tion metal compounds as catalysts (Yakobi, 1974). Acenaphtnene is stable
under laboratory conditions and resists photochemical degradation in soil
(Medvedev and Oavydow, 1972).
Laboratory experimentation points out the possibility of limited metaoo-
lism of acenaphthene to napthalic acid and napatholic anhydride.
A-l
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REFERENCES
Digurov, N.G., et al. 1970. Acenapnthequinone. Otkrytiya, Izobret. Prom.
Obraztsy, Tovarnyl Znaki. 47: 25. Rus.)
Lidner, R. 1931. Vapor pressures of some nydrocarbons. Jour. Phys. Cheml
35: 531 . '
Medvedev, V.A. and V.D. Oavydow. 1972. Transformation of individual coal
tar chemical industry organic products on chernozem soil. Pochvovedenie.
11: 22. (Rus.)
Menyailo, A.T., et al. 1971. 1,8-Naphthaldehyde carboxylic acid. Qtkry-
tiva, Izobret. Prom. Obraztsy, Tovarnyl Znaki. 48: 246. (Rus.)
Yakobi, V.A. 1974. Teor. Prakt. Zhrdkofazn. Okisneniva. 2nd ed. (Rus.)
A-2
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Aquatic Life Toxicology*
INTRODUCTION
The data base for acenaphthene and freshwater and saltwater organisms
is limted to a few acute toxicity tests under static conditions with un-
measured concentrations. A bioconcentration test has been conducted for 28
days and the depuration rate was determined. An embryo-larval test with the
sheepshead minnow has been conducted.
EFFECTS
Acute Toxicity
An acute test with Oaphnia magna resulted in a 48-hour ECS0 of 41,200
ug/1 and, when the bluegill was exposed to acutely lethal concentrations of
acenaphthene, the resulting 96-hour IC5Q value was 1,700 wg/l (U.S. EPA,
1S78) (Table •).
For the mysid shrimp (U.S. EPA, 1978) the 96-hour IC5Q is 970 ug/1,
and the 96-hour LC^g value for the sheepshead minnow is 2,230 «g/l (Table
1).
Chronic Toxicity
The acute-chronic ratio for the sheepshead minnow is small (3.1). The
96-hour IC5q was 2,230 ug/1 (Table 1) and the geometric mean of the noef-
fect and effect concentrations was 710 ug/1 (Table 2).
No other chronic data are available.
~The reader is referred to the Guidelines for Oeriving 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.
B-l
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Plant effects
The freshwater alga, Selenastrum caoricomutua, appears to be rather
sensitive with 96-hour EC^q values for chlorophyll £ and cell numbers of
530 and 520 ug/1, respectively (Table 3).
The saltwater alga* Sketetonema costatum, is more sensitive than the
sheepshead minnow and the mysid shrimp with a 96-hour EC5Q value for chlo-
rophyll a^ and cell numbers of 500 ug/1.
Residues
The bluegill accumulated acenaphthene during a 28-day exposure (U.S.
EPA, 1978) and the bioconcentration factor was 387 using ^C-acenaphthene
and thin-layer chromatography for verification (Table 4). The half-life of
this chemical in the whole body was less than 1 day.
Suwnary
The bluegill was much more sensitive to acenaphthene than the c 1 ado-
ceran, Daphnia magna: 50 percent effect concentrations are 1,700 and 41,200
ug/1, respectively. The freshwater alga, Selenastrum capricornutum. was
more sensitive than the fish species with a 96-hour ECS0 of 520 ug/1 for
cell number. The bioconcentration factor for the bluegill and acenaphthene
is 387 with a tissue half-life of less than 1 day.
Contrary to the pattern with freshwater species, the invertebrate spe-
cies, Mysidopsis bahla. was more sensitive (96-hour IC50 of 970 ug/1) than
the sheepshead minnow (96-hour LCjq of 2,230 ug/1). The saltwater alga,
Skeletonem costatum, was sensitive to acenaphthene with a 96-hour EC^ of
500 ug/1 for both chlorophyll _a and cell number. The acute-chronic ratio
for the sheepshead minnow is luite small (3.1) and indicates that successful
growth and reproduction occurs at a concentration close to one that causes
mortality.
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CRITERIA
The available data for acenaphthene indicate that acute toxicity to
i
freshwater aquatic life occurs at concentrations as low as 1,700 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
acenaphthene to sensitive freshwater aquatic animals but toxicity to fresh-
water algae occur at concentrations as low as 520 ug/1.
The available data for acenaphthene indicate that acute and chronic
toxicity to saltwater aquatic life occur at concentraztions as low as 970
and 710 ug/1. respectively, and would occur at lower concentrations among
species that are more sensitive than those tested. Toxicity to algae occurs
at concentrations as low as 500 ug/1.
B-3
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labia I. Icutt mIum lor KMiplitfcMM (U.S. CPA, 19711
Spaclat
Cladocaran,
Daflhnla aaqna
BIlMfllll,
Lapoalt ¦acrochlrus
MatHod"
IC50/EC50
JxatiL-
FRESHWATER SPECIES
S, U
S, U
41,200
1,700
Sfaclai Acuta
Valua tuo/ll
41,200
1,700
Mysld shrlap,
Myslfloptlt bah Ia
Shaapshaad alnnoM,
Cyprlnodon varlaqatut
SALTWATER SPECIES
S, U
S, u
970
2,230
970
2,230
• S • static, U - unaaaaurad
No Final Acuta Valuas ara calculable tine* tha alnlaua data taua
raqulraMnt& ara not Mt.
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09
I
u»
Ttblt 2. Chroalc valua* lor (MMpktkiM (U.^, LP A, INI)
Chroalc
Llalts Vain*
Spaclat Method1 (|ig/l) (Ma/I)
SALTWATER SPECIES
Sfcaapshaad alnnM, E-L 520-970 710
Cyprlnodow varlaqatus
1 E-t » aatryo-larval
Acute-Chronic Ratio
Chronic Acuta
Valua Valua
Spec Iat (Mfl/I) Ratio
ShaipshMd alnnov, 710 2,230 3*1
Cyprlnodon varlMAtus
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0
1
a\
Tabla I. PImI wIim for KMaplttlMM (U.S. CPA, lf?i|
Rata It
Spaclat Effact (mo/I)
FRESHWATER SPECIES
Alga, Chlorophyll a_ 530
Salanastrua caprIcornutua 96-lir EC50
Alga, Call number* 920
Salanastry caar Icornutua 96-hr EC&fl
SALTWATER SPECIES
Alga, Chlorophyllj» 500
Sk«latonatia costafua 96-hr EC50
Alga, Call counts 900
SKalatonama costatua 96-hr EC50
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Tabl* 4. RultfuM for aaumpkt*— (U.S. B»A, 1970)
BlocoHOMtratlon Ouratloa
Swclw TI»«w factor (Aim)
fR£StWATER SPECIES
BliMglll, Mho I* body 307 28
IXXMili wacrochlrus
0
1
-si
fr
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U?ER£NCES
U.S. EFA. 1978. In-depth studies on Health and environmental impacts c-
selected water pollutants. Contract No. 68-01-4646.
R_fl
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Mammalian Toxicology and Human Health Effects
EXPOSURE
Ingestion from Watec
Acenaphthene has been detected in the effluents from petro-
chemical, pesticide, and wood preservative industries by U.S. EPA
monitoring studies (U.S. EPA, 1978b). A survey of organic chemical
monitoring data from a variety of published and unpublished sources
indicated that acenaphthene had been identified in 11 studies (U.S.
EPA, 1976). Seven of these studies analyzed effluent from petro-
chemical or wood preserving plants, while two identified the chemi-
cal in finished drinking water, and another study found it in a
river sample. An analysis of the settling pond water from a wood
preserving plant showed acenaphthene present at a level of 0.2 mg/1
(U.S. EPA, 1973). Acenaphthene was also identified by two Russian
authors as one of several organic compounds found in wastewater as
a by-product of coke manufacturing (Andreikova and Kogan, 1977).
In an examination of water extracted by macroreticular resins
from a contaminated well in Ames, Iowa, investigators isolated ace-
naphthene at a level of 1.7 ppm (Burnham, et al. 1972). Identifi-
cation was verified by comparison with mass spectrum, chromatog-
raphy retention time, and ultraviolet spectrum of a standard. The
authors (Burnham, et al. 1972) noted that the contamination is
believed to be the result of residue from a coal gas plant which may
have leached into the aquifer after the plant closed in 1930.
Meijers and Van der Leer (1976) detected acenaphthene by gas
chromatography in a 20 liter sample of water from the river Maas ir
the Netherlands. Although not quantified by the authors, acenaph-
C-l
-------
tnen# was a minor constituent of the poiyeyciic aromatic hydro-
carbons (PAH) mixture identified in the water. Acenaphthene has a
low solubility in water, but.its presence in water may be signifi-
cant dye to possible adsorption on particulates.
Ingestion from Food
Only one study (Qnuska, et al. 1976) was found on the occur-
rence of acenaphthene in foods. Levels of >3.2 ug acenaphthene/kg
(the detection limit) were reportedly identified in the tissues of
shellfish of an unspecified species and location. Relative to
other PAH detected in this sample, the amount of acenaphthene was
small.
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 1CF for a lipid-soluble compound
in the tissues of various aquatic animals seem to be proportional
to the percent lipid in the tissue. Thus, the per capita ingestion
of a lipid-soluble chemical can be estimated from the per capita
consumption 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 per capita con-
sumption of freshwater and estuarine fish and shellfish in the
United States is 6.S g/day (Stephan, IStO). 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-2
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lipids for consumed freshwater and estuarine fish and shellfish, is
3.0 percent.
A measured steady-state BCF of 387 was obtained for acenaph-
thene using bluegills (U.S. EPA, 1978a). Similar bluegills con-
tained an average of 4.8 percent lipids (Johnson, 1980). Ar.
adjustment factor of 3.0/4.8 * 0.625 can be used to adjust the
measured SCF from the 4.8 percent lipids of the bluegill to the 3.0
percent lipids.that is the weighted average for consumed fish and
shellfish. Thus, the weighted average BCP for acenaphthene and the
edible portions of all freshwater and estuarine aquatic organisms
consumed by Americans is calculated to be 387 x 0.625 » 242.
Inhalation
Acenaphthene has been identified as one of many polycyclic
aromatic hydrocarbons (PAH) in gasoline exhaust condensate {Grim-
mer, et al. 1977) and cigarette smoke condensate (Harke, et al.
1976; Severson, et al. 1976). However, no estimates have been made
of the degree of exposure to acenaphthene that occurs to individ-
uals inhaling cigarette smoke or gasoline exhaust.
A 420,000 ft3 sample of air in Sydney, Australia, was found to
3
contain 3.9 ppm of solid acenaphthene, or 0.07 ug/100 m (Cleary,
1962), indicating that individuals in urban environments may be
exposed to measurable levels of acenaphthene'.
Dermal
Pertinent data could not be located in the available litera-
ture on dermal exposure to acenaphthene.
C-3
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PHARMACOKINETICS
Absorption and Distribution
Pertinent information could not be located in the available
literature on the absorption and distribution of acenaphthene.
Metabolism
Chang and Young (1943) isolated, by several methods, the anhy-
dride of naphthalene-1,8-dicarboxylic acid from the urine of two
groups of male white rats administered acenaphthene orally. One
group of rats was fed twice a day on a stock diet containing 1 per-
cent acenaphthene; a second group was dosed by gavage on alternate
days with 1 ml of a fine suspension of 0.1 g acenaphthene in dilute
starch solution. The authors suggested the possibility that the
naphthalic anhydride is a decomposition product of conjugated
metabolites that arose from the acid used in the extraction pro-
cedure, rather than a metabolic product of acenaphthene. Acenaph-
thene was not detected in the urine of the rats.
Aside from this study, no other data were found concerning the
metabolism of acenaphthene.
Excretion
Acenaphthene was not found in the acidified urine of rats
dosed orally with acenaphthene (Chang and Young, 1943). No other
data are available on the excretion of acenaphthene.
EFFECTS
Acute, Subacute, and Chronic Toxicity
Very little is known about the human toxicity of acenaphthene.
It is irritating to skin and mucous membranes, and may cause vomit-
ing if swallowed in large quantities (Sax, 1975).
C-4
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Similarly, limited data are available on the toxic effects of^
acenaphthene in mammals. Knobloch, et al. (1969) investigated the
acute and subacute toxic effects of acenaphthene in cats and mice.
Acenaphthene at 2 g/kg body weight administered orally in olive oil
to seven young rats (sex not specified) daily for 32 days caused
loss of body weight and changes in peripheral blood, increased
aminotransferase levels in blood serum, and produced mild morpho-
logical damage to both the.liver and kidney. A LD^q of 10 g/kg was
reported for rats and 2.1 g/kg for mice. The authors (Knobloch, et
al. 1969) noted that the morphological damage to the kidney and
liver was greater when acenaphthene was administered in a subacute
manner than when an acute dose was given. After 32 days of treat-
ment the animals showed mild bronchitis and localized inflammation
of the peribronchial tissue.
In another toxicity study, Reshetyuk, et al. (1970) exposed
100 rats to a 5-month chronic inhalation of acenaphthene at a level
of 12 + 1.5 mg/m^ for four hours a day, six days per week. Toxic
effects on the blood, lungs, and glandular constituents were
reported. The bronchial epithelium showed hyperplasia and meta-
plasia, which may have been symptoms of the pneumonia that killed a
large number of animals. However, no signs of malignancy appeared
during the 8-month post-exposure observation period. Reshetyuk, et
al. (1970) also reported a LD5Q of 600 + 60 mg/kg for rats given
intraperitoneal injections of acenaphthene. It must be pointed
out, however, that the lack of reported controls, as well as the
inadequate and confusing description of methods, make this study
unsuitable as the basis for a criterion.
C-5
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Gerjnbein (1975) investigated the effect of acenaphthene and
many other hydrocarbons upon the degree of liver regeneration in
partially hepatectomized male rats. Acenaphthene in peanut oil was
injected subcutaneously into one group of animals daily foe seven
days following surgery for a total dose of 5 to 20 mmol/kg. a
second group of animals was administered the chemical as part of
the diet at 0.03 and 0.10 percent (by weight). Ten days following
the surgical treatment, all animals were sacrificed and the liver
weights determined. Liver regeneration was significantly (p 0.01)
accelerated in both the injection-treated animals and the higher
oral dose group. A third group of rats was injected with acenaph-
thene three times and then sacrificed 72 hours after surgery.
Among all those exposed in this manner to five polycyclic hydrocar-
bons, acenaphthene-treated animals were the only animals showing a
significant acceleration of liver regeneration. These results are
in contrast to an earlier study by Gershbein (1958), in which a low
dose of 4.6 mmol/acenaphthene/kg did not result in a significant
liver regeneration acceleration. In the 1958 study, only a dose of
31.8 mmol/kg induced a significant regeneration.
Although the toxic effects of acenaphthene are not well docu-
mented, the reactions of humans to an odor from an aqueous solution
of the chemical, which may result in rejection of the contaminated
water, have been investigated. In a study of the odor thresholds
of organic pollutants (Lillard and Powers, 1975), a panel of 14
judges detected acenaphthene at a mean threshold of 0.08 ppm, with
a range of 0.02 to 0.22 ppm. Using these threshold values, extreme
value calculations were performed to predict levels of acenaphthene
C-6
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that a certain percentage of the population could detect. These
calculations ace shown as follows:
Percent of Population
Able to Detect Odor
Concentration of
Acenaphthene (ppm)
20
10
1
0.1
2.6 x 10"f
1.4 x 10",
1.9 x 10"^
2.1 x 10"4
Synergism and/or Antagonism
Two studies were conducted to investigate the effect of ace-
naphthene or the activity of dimethylnitrosamine demethyl'ase (DMN-
demethylase) , the liver en2yme that demethylates dimethylnitrosa-
mine (DMN), a known carcinogen. Argus, et al. (1971) and Arcos, et
al. (1976) injected male weanling rats intraperitoneally with ace-
naphthene at a concentration equimolar to 40 mg of 20-methylcholan-
threne/kg body weight. Twenty-four hours later, the animals were
sacrificed and the liver microsomes assayed for DMN-demethylase
activity. Acenaphthene showed a 0 percent (Argus, et al. 1971) and
a 5 percent (Arcos, et al. 1976) depression of the DMN-demethylase
levels over control rats with the same birth date. The difference
in enayme activity for the two studies may have been due to a modi-
fication of formaldehyde detection methods (Venkatesan, et al.
1968). Arcos, et al. (1976) noted that demethylation is a require-
ment foe carcinogenesis by DMN and, thus, it is possible that ace-
naphthAM nay slightly inhibit DMN carcinogenesis.
Buu-Hoi and Hien-Do-Phouc (1969) investigated the effect of
acenaphthene and other PAH on the activity of zoxazolamine hydroxy-
lase. Male" Wistar rats were injected intraperitoneally with
C-7
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20 rag/kg acenaphthene in corn ju, followed one week later by 90
mg/kg zoxazolaraine. The mean paralysis time of treated rats was
found to be significantly greater (pxrQ.Ol) than that of vehicle-
injected animals. The authors interpreted these results as an
indication that acenaphthene retards the detoxification of zoxazol-
amine, which ordinarily proceeds via hydroxylation.
Teratogenicity
Pertinent data could not be located in the available litera-
ture concerning the teratogenicity of acenaphthene.
Mutagenicity
The only data found on the mutagenicity of acenaphthene were
four studies using microorganisms as the indicator system (Clark,
1953a,b? Gibson, et al. 1978? Guerin, et al. 1978). No mutageni-
city was observed in any of the procedures used. Clark (1953a)
studied the effect of acenaphthene on the recombination rate of two
auxotrophic Escherichia coll strains. Acenaphthene was found to
have no appreciable effect upon the recombination rate of either
strain, as indicated by the low level of prototroph induction.
Acenaphthene did induce pleomorphism, but not the filamentous
"large" form which has been correlated with gene recombination. No
metabolic activation was used in this study and the dose of ace-
naphthene administered was not specified. In a later study, Clark
<1953b) tested acenaphthene for mutagenicity by exposing Micro-
coccus pyogenes var. aureus strain PDA209 to a saturated solution
of acenaphthene in a water-based nutrient broth without a metabolic
C-8
-------
activation system. When induction of mutants resistant to peni-
cillin or streptomycin was assessed, acenaphthene did not demon-
strate any mutagenic effects.
Two mutagenicity studies performed using Salmonella tvohimur-
ium gave negative or inconclusive results. Guerin, et al. (1978)
isolated an acenaphthene-containing aromatic subfraction from
shale-derived crude oil and tested it for mutagenicity using s.
typhimurium TA98. No increases were observed with or without rat
liver activation. Gibson, et al. (1978) exposed S. typhimurium
strains to 200 to 2,000 ug of acenaphthene dissolved in dimethyl-
sulfoxide after first irradiating the acenaphthene samples with
6G
Co to simulate (or replace) liver microsome activation. Unfortu-
nately, the results were erratic with major toxicity observed at
all dose levels tested. This toxicity obscured any assessment of
mutagenicity.
The studies discussed above were the only ones found in the
literature that examined the mutagenic potential of acenaphthene.
A' fifth study (Harvey and Balonen, 1968) examined the bindir. :f
acenaphthene to a variety of biologically important compound s
part of an unsuccessful attempt to correlate the nucleoside-bin-mg
activity of various chemicals with their carcinogenic potential.
Acenaphthene showed significant binding constants for caffeine and
riboflavin, but not for nucleosides.
Other Cellular Effects
The most thoroughly investigated effect of acenaphthene is its
ability to produce nuclear and cytological changes in microbial and
plant species. Most of these changes, such as an increase in cell
-------
- - ww»4w*=*iwf <***= wiwu u x i t uyt ion uc cne spindl6
mechanism during mitosis and the resulting induction of polyploidy.
While there is no known correlation between these effects and the
biological impact of acenaphthene on mammalian cells, these effects
are reported in this document because they are the only substan-
tially investigated effects of acenaphthene.
Ten experiments examining the effect of acenaphthene on plants
and eight others involving the effects upon microorganisms are dis-
cussed in the following sections. A summary of these data is pre-
sented in Table 1.
Plants: Kostoff (1938a) exposed Nicotiana longiflora shoots
to vapor from acenaphthene crystals and examined the shoots for
effects on mitosis and/or meiosis. The exposure induced tetraploid
and octaploid shoots, which produced seeds of new polyploid plants.
The polyploidizing effect of acenaphthene vapor increased with in-
creases in the length of exposure or the number of particles used.
Kostoff (1938b) also tested the effect of acenaphthene on the
branches of floral buds of nine Nicotiana species. Meiosis in the
buds proceeded abnormally also, with the bivalent chromosomes fail-
ing to arrange correctly on the equatorial plate. They tended to
spread into the cytoplasm singly or in groups, resulting in a vari-
able number of chromosomes per nuclei at the end of the second
division. Fifty to one-hundred percent of the pollen produced by
the end of Miosis was abortive.
In th« same study, Kostoff (1938b) covered germinating seeds
from a variety of plants with acenaphthene crystals to study the
effects on mitosis. Cereals and grasses (wheat, rye, barley, oat,
C-10
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TABLE 1
Summary of Polyploid and Other Mitotic Effects Induced
by Acenaphthene in Plants uiid Microorganisms
Organism
Treatment
Effects Noted
Reference
Plants:
Nicotiana shoots
Vapor
Stable polyploidy;
abnormal, abortive
meiosis
Kostoff,
1938a,b
Cereal, grass
legume, and
compositae seeds
Cherry-mazzard
hybrid seeds
Allium cepa L.
Allium cepa L.,
A. sativum
Allium fistulosum,
Colchicum roots
Allium root cells
Crystals
(4-12 days)
Powder
(10 hours)
Saturated
solut ion
(2-5 days)
Treatment
unspecified
Crystals wrapped
in moist filter
paper (4-20 days)
Vu
(12-ju hours)
Abnormal mitosis,
spindle mechanism
inhibited
Seed germination and
growth inhibited; no
polyploidy
Chromosome
fragmentation,
polyploidy
Frequency of division
retarded, multiple
prophase
C-mitosis, polyploidy,
root-tip swellings
Random cell wall
development
Kostoff, 1938b
Zhukov, 1971
D'Amato, 1949
Mookerjee, 197
Levan, 1940
Mesquita, 1967
-------
TABLE 1 (continued)
Organism
Treatment
Effects Noted
Reference
Binucleate pollen
Tradescantia
pollen
Tradescantia
stamen hairs
Fungi:
Basidoroycetes
Basidiobolus
ranarum hyphae
Pythium
aphanidermature
hyphae
Yeast
Candida scottii
Vapor
Vapor
Saturated solution
(2-4.5 hours)
Vapor
Vapor
(6-18 hours)
Vapor or
supersaturated
solution
(12 hours)
10"1 to
3 x 10~ mol
solution
0.2-1.0%
agar
Spindle inhibited,
division stopped
at metaphase
Spindle disturbed
No polyploidy, no
chromosomes in
metaphase
Mitotic frequency
decrease; growth,
pigment formation,
differentiation and
morphology changes
Alterations in
nuclear division
Nuclear division
arrested; pyknosis
No lethality or
c-mitosis
Increase in cell
size, nucleus,
DNA content
Dyer, 1966
Swanson, 1940
Nebel, 1938
Hoover, 1972
Hoover and
Liberta, 1974
Seshadri and
Payak, 1970
Levan and
Sandwall, 1943
Iroshenetsky,
et al. 1966
-------
TABLE 1 (continued)
Organism
Treatment
Effects Noted
Reference
Bacteria:
Mycobacterium
rubrum
Rhizobium
Algae:
Chara globular is;
Mite11a
flagelliformis
Vapor from
10-20 mg
crystals
Vapor
Saturated
solution
(12-120 hours)
Elongation and
thickening of
cells; unstable
polyploidy
Increase in DNA
content; change in
biochemical
properties
Number of cells in
mitosis reduced;
chromosomes clumped
at metaphase;
chromosomes doubled
.Imshenetsky
and Zhil'tsova,
1973
Avvakumova,
et al. 1975
Sarma and
Tr ipathi,
1976a,b
-------
-------
maize, and rice) showed slow growth and abnormal roots and leaf
formations after 4 to 8 days. Legumes evidenced the.se effects
after 6 to 12 days, while Compositae reacted in a time period mid-
way between the other two groups. Mitosis in these seedlings pro-
ceeded abnormally; the spindle mechanism was inhibited and the
chromosomes were not arranged on the equatorial plate. Failure of
the chromosomes to move to the poles resulted in polyploidy.
Zhukov (1971) investigated the effect of acenaphthene on plant
seeds. He treated "cherry-mazzard hybrid" seeds with acenaphthene
powder for 10 hours. Seed germination and seedling growth were
inhibited, but r.o polyploidal cells were found in the plant roots.
Four investigators performed experiments with acenaphthene
and Allium plants. When treated with saturated solutions of ace-
naphthene in either tap or distilled water for 2 to 5 days, Allium
cepa demonstrated intense chromosome fragmentation (D'Amato,
1949) . Fragmenting effects on diploid and polyploidized nuclei in
the resting, stage were noted, as were centromere effects on the
metaphase chromosomes and, occasionally, on chromatids at anaphase.
In a later study (Mookerjee, 1973) , acenaphthene exposure (concen-
tration unspecified) was found to retard the frequency of division
of Allium cepa and Allium sativum. Multiple prophase was observed
in A. cepa.
L«v«fi (1940) dusted Allium fistulosum and Colchicum roots with
acenaphthene crystals and then wrapped the plants in moist filter
paper. After four days of growth, the spindles were altered and
the centromeres inactivated: this process has been termed
"c-mitosis" because a similar effect occurs with colchicine treat-
C-14
-------
ment. Tetraploid and octaploid cells were formed within 14 to 20
days, resulting in the formation of root-tip swellings (c-tumors)
in Allium. Mesquita (1967) also investigated the effects of ace-
naphthene on Allium root cells. He exposed A. cepa root tips to
acenaphthene vapor at room temperature for 12 to 96 hours. The re-
assembling of the phragmoplast elements (small pieces of the endo-
plasmic reticulum and Golgi bodies) in the equatorial region was
inhibited, but the fusion of these elements in other parts of the
cell was unimpaired. The result was the random development of cell
walls.
To investigate the effect of acenaphthene on mitosis, Oyer
(1966) exposed plant species with binucleate pollen (such as Bel-
levalia romana, Tulbaghia natalensis, and Antirrhinum majus) to
vapor from acenaphthene crystals. He found that all cells remained
at metaphase, with anaphase being inhibited due to an inhibition of
the mitotic spindle. Swanson (1940) also observed effects on mito-
sis in plant pollen. He scattered acenaphthene crystals on the
bottom of a petri dish in which Tradescantia pollen was incubated.
The vapors acted by disturbing the spindle mechanism so that the
chromosomes remained in place after division. Nebel (1938) exam-
ined the effect of acenaphthene on mitosis in plant hairs by treat-
ing stamen hairs of Tradescantia with a saturated solution of ace-
naphth«n« in liquid media for 2 and 4.5 hours. He found no poly-
ploid cells and no nuclei showing chromosomes in a metaphase con-
dition.
Microorganisms: Several experiments have been performed to
investigate the effect of acenaphthene on microorganisms. Hoover
C-15
-------
(1972) exposed 37 species of Basidiomycetes to acenaphthene vapors
or media containing acenaphthene at unspecified dose levels in
order to examine effects on growth, pigment, morphology, nuclear
division, and fruit body formation. As the treatment time in-
creased, changes in nuclear division, became more pronounced, with a
concurrent decrease in the mitotic frequency. Growth, pigment for-
mation, differentiation, and colonial and cellular morphology were
affected by acenaphthene treatment. A delay or prevention of
light-induced fruitbody formation occurred in one species; two spe-
cies developed greatly enlarged fruitbodies as a result of this
treatment. The genetic stability o£ these phenotypic changes was
not demonstrated, however.
In a later experiment, Hoover and Liberta (1974) exposed
hyphae cultures of the fungus Basidiobolus ranarum to acenaphthene
vapor for 6 to 18 hours. At the end of 18 hours, gross alterations
in nuclear division were observed and the spindle fibers were ren
dered unstair ole. The time required for division was significar
ly increased in acenaphthene-treated cells. The effect of acenf
thene on fungi was also investigated by Seshadri and Payak (1'
They exposed hyphae of Pythium aphanidermatum to acenaphthene
or to a supersaturated solution of acenaphthene for 12 hour
apor proved instrumental in arresting the progress of
division. A marked increase in the size and number of
nuclei was noted, and the nuclei showed various degrees <
and shape irregularity.
Levan and Sandwall (1943)- examined the effect of
centrations of acenaphthene (1 x 10"1 to 3 x 10~7
C-16
-------
ethanol) on wort yeast cell cultures. Even at the highest concen-
tration/ there was no lethality or effect on cell propagation. The
authors concluded that the c-mitotic action demonstrated by ace-
naphthene in higher plants was not observable in yeast. Polyploidy
was induced, however, in the yeast Candida scottii (a yeast without
a sexual cycle) when treated with 0.2 percent and 1.0 percent ace-
naphthene added to agar medium (Imshenetsky, et al. 1966). The
size of the cell and the nucleus were both increased in the treated
cultures, and there was also a higher dry biomass for these cells.
The DNA content (ug per cell) was higher in acenaphthene-treated
cells, although the difference between experimental and control
cultures decreased as the cultures aged.
Imshenetsky and Zhil'tsova (1973) attempted to produce "poly-
ploid-like" cells by exposing Mycobacterium rubrum to vapor from 10
to 20 rag acenaphthene. When the vapor was used alone for treat-
ment, there was no increase in the size of the cells, nor any indi-
cation of the induction of polyploidy. When the cells were treated
with water or ethylenediaminetetraacetic acid (EDTA) to increase
membrane permeability, acenaphthene vapor treatment caused elonga-
tion and thickening of cells, with a longer development cycle;
these "polyploid-like" changes were found to be unstable, however.
In another experiment with bacteria, Avvakumova, et al. (1975)
treated Rhigobium (nodule-forming bacteria) with acenaphthene
vapor (dose unspecified) to induce polyploidy. The authors found
an acenaphthene-associated increase in cellular DNA content and
biomass, as well as a change in biochemical properties, e.g., the
ability to assimilate carbohydrates and/or organic acids.
C-17
-------
Acenaphthene has also been shown to affect mitosis in two spe-
cies of algae. Sarma and Tcipathi (1976a,b) treated Chara glo-
bular is and Nitella flagelliformis with a saturated solution of
acenaphthene for 12 to 120 hours. The number of cells in mitosis
was reduced by 40 percent, and the chromosomes were seen to clump
at metaphase after 120 hours. Nine percent of the C. globular is
cells showed complete chromosome doubling by the end of the treat-
ment period.
Carcinogenicity
Very little work has been done to determine whether acenaph-
thene may have carcinogenic properties. Heukomm (1974) reported
negative results in a predictive test for carcinogenicity based
upon neoplastic induction in the newt Triturus cristatus. Ten ani-
mals were injected subcutaneously with acenaphthene (dose and sol-
vent not reported) in the fleshy part of the tail along the verte-
bral axis. Samples of the injection site were removed at 7 and 14
days, and the tissues were examined for neoplastic infiltration in
the epidermis and the development or regression of diffuse tumors.
Neoplastic lesions were divided into three categories depending on
the size of the lesion and assigned a numerical coefficient accord-
ingly: large (1.0), intermediate (0.5), and limited (0.25), Cal-
culation of a neoplastic index by summing the coefficients of all
lesions and dividing by the number of observed animals gave an
index for acenaphthene of 0.0, indicating a lack of neoplastic
induction in the newt.
Neukomm (1974) discussed the reliability of this test by draw-
ing a correlation between positive index values for a few polycy-
C-18
-------
clic aromatic hydrocarbons and the carcinogenicity of these same
compounds for mouse skin. These limited comparisons, however, are
not sufficient to establish the value of this test for predicting
carcinogenicity in mammalian systems.
The only, other carcinogenicity studies in the literature in-
volving acenaphthene considered it as one component of a complex
mixture of PAH. It is impossible in these studies to sort out the
relative contribution of acenaphthene versus other hydrocarbons in
the mixture, so no real conclusions can be drawn. Akin, et al.
(1976) isolated some polycyclic hydrocarbon-rich fractions of the
neutral portion of cigarette smoke condensate (CSC) and tested them
for tumor promotion on female mouse skin, using 7,12-dimethy,lbenz-
(a)anthracene (DMBA) as the initiator. Animals were painted once
with 125 ug DMBA on dorsal skin; 3 to 4 weeks later the fractions
were applied five times a week for 13 months. The fraction con-
taining acenaphthene, pyrene, phenanthrene, and other PAH, showed
no significant tumor-promoting activity over controls treated with
DMBA and acetone. This result was surprising in view of the fact
that Scribner (1973) had demonstrated the tumor-promoting ability
of pyrene and phenanthrene.
In 1962, Hoffman and Wynder found that benzene extracts of
gasolin® exhaust condensates were carcinogenic in mouse skin paint-
ing tests. This study is of interest considering a later study by
Grimmer, «t al. (1977) which showed that acenaphthene was present
in an unspecified concentration in the benzene extracts of gasoline
C-19
-------
exhaust condensate. Unfortunately, the possible contribution
acenaphtherie'to the observed carcinogenicity (Hoffman and Wynde
1962) cannot be determined from this limited evidence.
C-20
-------
CRITERION FORMULATION
Existing Guidelines and Standards
No 'existing guidelines or standards were found.
Current Levels of Exposure
Virtually no information is available concerning the preva-
lence or concentration of acenaphthene in the environment. Ace-
naphthene has been detected in cigarette smoke (Harke, et al. 1976;
Severson, et al. 1976), automobile exhaust (Grimmer, et al. 1977),
and in urban air (Cleary, 1962) and is present in coal tar and sev-
eral fossil fuel' oils. It has also been reported in wastewater
crom petrochemical, pesticide, and wood preservative industries
(U.S. EPA, 1978b) and detected in water from a river in the Nether-
lands (Meijers and Van der Leer, 1976).
Special Groups at Risk
Individuals working with coal tar and/or its products face a
possible risk due to increased exposure to acenaphthene, although
no data are available to estimate this risk.
Ba3i3 and Derivation of Criterion
So little research has been performed on acenaphthene that its
mammalian and human health effects are virtually unknown. The two
toxicity studies available (Knobloch, et al. 1969; Reshetyuk, et
al. 1970} ace inadequate for use as the basis of a criterion due to
deficiencies in the experimental designs (lack of controls, small
number of animals, short durations, etc.). Therefore, until more
toxicological data are generated, particularly on genotoxic ef-
fects, a criterion based upon organoleptic data is proposed. The
lowest levels eliciting human responses were reported to be 0.022
C-21
-------
to 0.22 ppm (Lillard and Powers, 1975). Thus, the lower limit 0.02
ppm (0.02 mg/1) appears to be the best estimate of a criterion
level that will prevent unpleasant odoc from acenapthene. It is
emphasized that this criterion is based on aesthetic considerations
only and as such has no demonstrated relationship to potential
adverse human health effects.
Since the recommended criterion is based on organoleptic ef-
fects and is not a toxicological assessment, the consumption of
fish and shellfish products was not considered as a route of expo-
sure.
This criterion will be reviewed when additional toxicological
data are available.
C-22
-------
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