ACENAPHTHENE
Ambient Water Quality Criteria
Criteria and Standards Division
Office of Water Planning and Standards
U.S. Environmental Protection Agency
Washington, D.C;
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CRITERION DOCUMENT
ACENAPHTHENE
CRITiERIA
Aquatic Life
The data base for freshwater aquatic life is insufficient to
allow use of the Guidelines. The following recommendation is
inferred from toxicity data for saltwater organisms.
For acenaphthene the criterion to protect freshwater aquatic
life as derived using procedures other than the Guidelines is 110
ug/1 as a 24-hour average and the concentration should not exceed
240 ug/1 at any time.
For acenaphthene the criterion to protect saltwater aquatic
life as derived using the Guidelines is 7.5 ug/1 as a 24-hour
average and the concentration should not exceed 17 ug/1 at any
time.
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Human Health
For the prevention of adverse effects due to the organoleptic
properties of acenaphthene in water, the criterion recommended is
.02 mg/1.
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Introduction
Acenaphthene (1,2-dehydro-acenaphthylene 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 polynuclear aromatic hydrocarbon with a molecular weight
of 154 and a formula of
The compound is a white crystalline solid at room tem-
perature with a melting range of 95 to 97°C and a boiling
range of 278 to 280°C (Lidner, 1931). The vapor pressure
is less than 0.02 mm Hg. Acenaphthene is soluble in water
(100 mg/1), but solubility increases in organic solvents
such as ethanol, toluene, and chloroform.
Acenaphthene will react with molecular oxygen in the
presence of alkali-earth metal bromides to form acenaphthequinone
(Digurov, et al. 1970). In the presence of alkali-earth
metal hydroxides, acenaphthene reacts with ozone to produce
1,8-naphthaldehyde carboxylic acid (Menyailo, et al. 1971).
Acenaphthalene can be oxidized to aromatic alcohols and
ketones using transition metal compounds as catalysts (Yakobi,
1974). Acenaphthene is stable under laboratory conditions
and resists photochemical degradation in soil stability
studies (Medvedev and Davydow, 1972).
Acenaphthene has been demonstrated to affect the growth
of plants through improper nuclear division and polypoidal
chromosome number. These same observations were noted in
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several microorganisms as well. Little information regarding
aquatic toxicity was found. The freshwater acute value
for bluegill was 1,700 pg/1, and the bioconcentration factor
was 397. Saltwater toxicity to the sheepshead minnow was
2,230 ^jig/1, and no bioconcentration data were available.
Data on toxicity to non-human mammals were few and virtually
no incidences of human acenaphthene toxicity were noted.
There was some information found showing organoleptic effects
attributed to acenaphthene in water. A detection range
of 0.02 to 0.22 mg/1 was given. Laboratory experimentation
points out the possibility of limited metabolism of acenaph-
thene to napthalic acid and napatholic anhydride.
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REFERENCES
Digurov, N.G., et al. 1970. Acenaphthequinone. Otkrytiya,
Izobret. Prom. Obraztsy, Tovarnyl Znaki. 47: 25.
Lidner. 1931. Vapor pressures of some hydrocarbons. Jour.
Phys. Chem. 35: 531.
Medvedev, V.A., and V.D. Davydow. 1972. Transformation
of individual coal tar chemical industry organic products
on chernozem soil. Pochvovedenie 11: 22.
Menyailo, A.T., et al. 1971. 1,8-Naphthaldehyde carbozylic
acid. Otkrytiva, Izobret. Prom. Obraztsy, Tovarnyl Znaki.
48: 246.
Yakobi, V.A. 1974. Teor. Prakt. Zhrdkofazn. Okisheniva.
2nd ed. Moscow.
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AQUATIC LIFE TOXICOLOGY*
FRESHWATER ORGANISMS
Introduction
The data base for acenaphthene and freshwater organisms is
limited to a few acute toxicity tests under static conditions with
unmeasured concentrations and no criterion can be derived from
these results. A bioconcentration test has been conducted for 28
days and the depuration rate was determined.
Acute Toxicity
The bluegill has been exposed to acutely lethal concentra-
tions of acenaphthene (U.S. EPA, 1978) and the resulting adjusted
96-hour LC50 value is 929 ug/1 (Table 1). After use of the
sensitivity factor (3.9), this result leads to the Final Fish
Acute Value of 240 ug/1.
An acute test with Daphnia magna resulted in an adjusted
48-hour EC50 of 34,900 ug/1 (Table 2) and the Final Invertebrate
Acute Value derived from that datum is 1,700 ug/1. Since the
*The reader is referred to the Guidelines for Deriving Water
Quality Criteria for the Protection of Aquatic Life [43 FR 21506
(May 18, 1978) and 43 FR 29028 (July 5, 1978)3 in order to better
understand the following discussion and recommendation. The
following tables contain the appropriate data that were found in
the literature, and at the bottom of each table are the calcula-
tions for deriving various measures of toxicity as described in
the Guidelines.
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equivalent value (240 ug/1) for fish is lower, it becomes the
Final Acute Value.
Chronic Toxicity
No chronic tests have been reported for acenaphthene and
freshwater organisms.
Plant Effects
The alga, Selenastrum capricornutum, appears to be rather
sensitive. The 96-hour EC50 values for chlorophyll a and cell
numbers are 530 and 520 ug/1* respectively (Table 3). The Final
Plant Value is 520 ug/1.
Residues
The bluegill accumulated acenaphthene during a 28-day
exposure (U.S. EPA, 1978). The bioconcentration factor was 387
using 14C-acenaphthene and thin-layer chromatography for
verification (Table 4). The half-life of this chemical in the
whole body was less than 1 day.
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:riterion formulation
Freshwater Aquatic Life
Summary of Available Data
The concentrations below have been rounded to two significant
figures.
Final Fish Acute Value = 240 ug/1
Final Invertebrate Acute Value = 1,700 ug/1
Final Acute Value = 240 ug/1
Final Fish Chronic Value = not available
Final Invertebrate Chronic Value = not available
Final Plant Value = 520 ug/1
Residue Limited Toxicant Concentration = not available
Final Chronic Value = 520 ug/1
0.44 x Final Acute Value = 110 ug/1
No freshwater criterion can be derived for acenaphthene using
the Guidelines because no Final Chronic Value for either fish or
invertebrate species or a good substitute for either value is
available.
Results obtained with acenaphthene and saltwater organisms
indicate how a criterion may be derived.
For acenaphthene and saltwater organisms, 0.44 times the
Final Acute Value is less than the Final Chronic Value which is
derived from results of an embryo-larval test with the sheepshead
minnow. Therefore, it seems reasonable to estimate a criterion
for acenaphthene and freshwater organisms using 0.44 times the
Final Acute Value.
The maximum concentration of acenaphthene is the Final Acute
Value of 240 ug/1 and the 24-hour average concentration is 0.44
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times the Final Acute Value. No important adverse effects on
freshwater aquatic organisms have been reported to be caused by
concentrations lower than the 24-hour average concentration.
CRITERION: For acenaphthene the criterion to protect fresh-
water aquatic life as derived using procedures other than the
Guidelines is 110 ug/1 as a 24-hour average and the concentration
should not exceed 240 ug/1 at any time.
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Table 1. Freshwater fish acute values for acenaphthene (U.S. EPA, 1978)
Adjusted
Bioaeeay Test Time LC50 LC50
Organism Method" Cone.** thra) (uq/l> (ug/H
Blueglll, S U 96 1,700 929
Lepomis macrochlrus
* S = static
** U = unmeasured
Ceometric mean of adjusted values
- 929 ng/1 - 240 ng/1
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TabXe 2. Fcesfawater invertebrate acute values for accnaphthene (U.S. EPA,
Adjusted
fJicassay Test Time LC50 IC!>U
Oi ti.inisin Meti.ou'^ Cone .** (ins) (u<|/ H (u
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. t
Table 3. Freshwater plcmt effects for acenaphthene (U.S. EPA, 1978)
Organism
Effect
Concentration
(ug/l>
Alga,
Sclenaatrum
caprlcornutum
Alga,
Selenastrum
caprlcornutum
96-hr 1£C50 530
chlorophyll a
96-hr EC50 520
cell numbers
Lowest plant value 0 520 Mg/1
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Table A. Freshwater residues for acenaphthene (U.S. EPA, 1978)
Time
Orgjniain bioconcentrdtion t'actot (days)
Bluegill, 387 28
Laponiis macroch 1 rus
DO
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SALTWATER ORGANUAA)
Introduction
As with freshwater organisms there is a limited data base for
acenaphthene and saltwater organisms. There is little difference
between LC50 and EC50 values for the sheepshead minnow, Cyprinodon
variegatus, the mysid shrimp, Mysidopsis bahia, or the alga,
Selenastrum costatum. An embryo-larval test has been conducted
with the sheepshead minnow.
Acute Toxicity
The adjusted 96-hour LC50 value for the sheepshead minnow is
1,219 ug/1 (Table 5). Based on this datum, the Final Fish Acute
Value for acenaphthene and saltwater fish is 330 ug/1.
For the mysid shrimp (U.S. EPA, 1978) the 96-hour LC50 is 821
ug/1 (Table 6). The Final Invertebrate Acute Value is 17 ug/1*
and, since this concentration is lower than the comparable value
for fish, it also becomes the Final Acute Value.
Chronic Toxicity
The ratio of acute and embryo-larval test results with the
sheepshead minnow is small. The unadjusted 96-hour LC50 was 2,230
ug/1 (Table 5) and the geometric mean of the no-effect and effect
concentrations was 710 ug/1 (Table 7). The chronic value, de-
rived by dividing this geometric mean by 2, is 355 ug/1- When
this concentration is divided by the sensitivity factor (6.7), the
Final Fish Chronic Value of 53 ug/1 is obtained.
Plant Effects
As discussed earlier, the alga, Skeletonema costatum, is as
sensitive as the sheepshead minnow and the mysid shrimp. The
96-hour EC50 numbers is 500 ug/1.
This also is
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CRITERION FORMULATION
Saltwater Aquatic Life
__ | M..Ill -¦**¦- -- ¦' mi
Summary of Available Data
The concentrations below have been rounded to two significant
figures.
Final Fish Acute Value = 330 ug/1
Final Invertebrate Acute Value = 17 ug/1
Final Acute Value = 17 ug/1
Final Fish Chronic Value » 53 ug/1
Final Invertebrate Chronic Value = not available
Final Plant Value = 500 ug/1
Residue Limited Toxicant Concentration = not available
Final Chronic Value = 53 ug/1
0. 44 x Final Acute Value = 7.5 ug/1
The maximum concentration of acenaphthene is the Final Acute
Value of 17 ug/1 and the 24-hour average concentration is 0.44
times the Final Acute Value. No important adverse effects on
saltwater aquatic organisms have been reported to be caused by
concentrations lower than the 24-hour average concentration.
CRITERION: For acenaphthene the criterion to protect salt-
water aquatic life as derived using the Guidelines is 7.5 ug/1 as
a 24-hour average and the concentration should not exceed 17 ug/1
at any time.
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Table 5. Marine fish acuce values for acenaphthene (U.S. EPA, 1978)
Adjusted
Bioaaaay Test Time LC50 LC50
Organlam Method* Cone.** (hra) (ug/1) (ug/ll
Sheepshead minnow, S U 96 2,230 1,219
Cyprinodon variegatus
* S = static
** U = unmeasured
1 219
Geometric mean of adjusted values - 1,219 ug/1 ' 7' a Pg/1
4
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Table 6. Marine invertebrate acute values for atenaphthene (U.S. EPA, 1978)
Adjusted
Qiodssay Ttst fune I.C50 I.C'jI)
Oman ism Mfetnoa * Cor.c. ** Has) tug/Q t"
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'faille 7. Marine fish chronic values for acenaphthene (U.S. EPA, 1978)
Chronic
Limits Value
Organism Test*
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Table 8. Marine plane effects for acenaphlhene (U.S. EPA, 1978)
Concentration
Organism Effect (uq/l>
Alga,
Skfcieconema costatum
Alga,
Skeletonema costatum
EC50 96-hr 500
chlorophyll a
EC50 96-hr 500
cell counts
Lowest plant value a 500 )>g/l
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REFERENCES
U.S. Environmental Protection Agency. 1978. In-depth studies
on health and environmental impacts of selected water pollu-
tants. U.S. Environ. Prot. Agency. Contract No. 68-01-4646.
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Mammalian Toxicology and Human Health Effects
EXPOSURE
Ingestion from Water
Acenaphthene has been detected in the effluent from
petrochemical, pesticide, and wood preservative industries
by U.S. Environmental Protection Agency 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 petrochemical, or wood preserving plants, while two
identified the chemical 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 in the wastewater from
by-product coke manufacture (Andreikova and Kogan, 1977).
In an examination of water extracted by macroreticular
resins from a contaminated well in Ames, Iowa, investigators
isolated acenaphthene at a level of 1.7 ppm (Burnham, et
al. 1972). Identification was verified by comparison with
mass spectrum, 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
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20-liter sample of water from the river Maas in the Nether-
lands. Although not quantified by the authors, acenaphthene
was a minor constituent of the PAH mixture identified in
the water. Acenaphthene has a low solubility in water,
but its presence in water may be significant due to possible
adsorption on particulates.
Ingestion from Foods
Only one study (Onuska, et al. 1976) was found on the
occurrence of acenaphthene in foods. Levels of J> 3.2 pg
acenaphthene/kg (the detection limit) were presumptively
identified in the tissues of shellfish of an unspecified
species and location. Relative to other PAHs detected in
this sample, the amount of acenaphthene was small.
A bioconcentration factor (BCF) relates the concentra-
tion of a chemical in water to the concentration in aquatic
organisms, but BCF's are not available for the edible por-
tions of all four major groups of aquatic organisms consumed
in the United States. Since data indicate that the BCF
for lipid-soluble compounds is proportional to percent lipids,
BCF's can be adjusted to edible portions using data on percent
lipids and the amounts of various species consumed by Americans.
A recent survey on fish and shellfish consumption in the
United States (Cordle, et al. 1978) found that the per capita
consumption is 18.7 g/day. From the data on the nineteen
major species identified in the survey and data on the fat
content of the edible portion of these species (Sidwell,
et al. 1974), the relative consumption of the four major
groups and the weighted average percent lipids for each
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group can be calculated:
Consumption Weighted Average
Group
(Percent)
Percent Lipids
Freshwater fishes
12
4.8
Saltwater fishes
61
2.3
Saltwater molluscs
9
1.2
Saltwater decapods
18
1.2
Using the percentages for consumption and lipids for each
of these groups, the weighted average percent lipids is
2.3 for consumed fish and shellfish.
A measured steady-state bioconcentration factor of
387 was obtained for acenaphthene using bluegills containing
about one percent lipids (U.S. EPA, 1978a). An adjustment
factor of 2.3/1.0 =2.3 can be used to adjust the measured
BCF from the 1.0 percent lipids of the bluegill to the 2.3
percent lipids that is the weighted average for consumed
fish and shellfish. Thus, the weighted average bioconcentra-
tion factor for acenaphthene and the edible portion of all
aquatic organisms consumed by Americans is calculated to
be 387 x 2.3 = 890.
Inhalation
Acenaphthene has been identified as one of many poly-
cyclic aromatic hydrocarbons (PAHs) in gasoline exhaust
condensate (Grimmer, et al. 1977) and cigarette smoke conden-
sate (Harke, et al. 1976; Severson, et al. 1976). However,
no estimates have been made of the degree of exposure to
acenaphthene that occurs to individuals inhaling cigarette
smoke or gasoline exhaust.
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A 420,000 cubic foot sample of air in Sydney, Australia,
was found to contain 3.9 ppm of solid acenaphthene, or 0.07
pg/100 m3 (Cleary, 1962), indicating that individuals in
urban environments may be exposed to measurable levels of
acenaphthene.
Dermal
No information is available on dermal exposure to ace-
naphthene .
PHARMACOKINETICS
Absorption *
No data are available on the absorption of acenaphthene.
Distr ibution
No data are available on the distribution of acenaph-
thene.
Metabolism
Chang and Young (1943) isolated, by several methods,
the anhydride 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 percent 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 raised the possibility that
the naphthalic anhydride is a decomposition product of con-
jugated compounds arising from the acid used in the extrac-
tion procedure, rather than a metabolic product of acenaph-
thene. No acenaphthene was detected in the urine of the
rats.
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Aside from this study, no other data were found con-
cerning the metabolism of acenaphthene.
Excretion
As indicated previously, no acenaphthene was found
in the acidified urine of rats dosed orally with acenaph-
thene (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 ace-
naphthene. It is irritating to skin and mucous membranes,
and may cause vomiting if swallowed in large quantities
(Sax, 1975).
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 ace-
naphthene in rats and mice. Two g acenaphthene per kg body
weight administered orally in olive oil to seven young rats
(sex not specified) on a daily basis for 32 days caused
loss of body weight, changes in peripheral blood, heightened
aminotransferase levels in blood serum, and mild morphological
damage to both the liver and kidney. An LD^g 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 the animals showed mild bronchitis
and localized inflammation of the peribronchial tissue.
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In another toxicity study, Reshetyuk, et al. (1970)
exposed 100 rats to a five-month chronic inhalation of acenaph-
thene at a level of 12+1.5 mg/m3 for four hours a day, six
days per week. Toxic effects to the blood, lungs, and glandular
constituents were reported. The bronchial epithelium showed
hyperplasia and metaplasia, possibly a symptom of the pneu-
monia which killed a large number of animals. However,
no signs of malignancy appeared during the 13 months of
exposure. Reshetyuk, et al. (1970) also reported an LD^g
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 unsuit-
able as the basis for a criterion.
Gershbein (1975) investigated the effect of acenaphthene
and many other hydrocarbons upon the degree of liver regenera-
tion in partially hepatectomized male rats. Acenaphthene
in peanut oil was injected subcutaneously into one group
of animals on a daily basis for seven days following surgery
for total dose of 5 to 20 mM/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 were injected
with acenaphthene three times and then sacrificed 72 hours
after surgery. Among all those exposed in this manner to
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5 polycyclic hydrocarbons, 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 mM acenaphthene per kg did not result in a significant
liver regeneration acceleration. In the 1958 study, only
a dose of 31.8 mM/kg induced a significant regeneration.
Although the toxic effects of acenaphthene are not
well documented, 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 .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 acenaph-
thene that a certain percentage of the population could
detect. These calculations are shown below:
Percent of Population
Able to Detect Odor
20
10
1
0.1
Synergism and/or Antagonism
Two studies were conducted to investigate the effect
of acenaphthene on the activity of dimethylnitrosamine deme-
thylase (DMN-demethylase), the liver enzyme that demethylates
DMN, a known carcinogen. Argus, et al. (1971) and Arcos,
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Concentration of
Acenaphthene (ppm)
2.6 x 10"?
1.4 x 10 ^
1.9 x 10 4
2.1 x 10"4
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et al. (1976) injected male weanling rats intraperitoneally
with acenaphthene at a concentration equimolar to 40 mg
of 20 methylcholanthrene/kg body weight. Twenty-four hours
later, the animals were sacrificed and the liver microsomes
assayed for DMN-demethylase activity. Acenaphthene showed
a zero (Argus, et al. 1971) and a five percent (Arcos, et
al. 1976) repression of the DMN demethylase levels over
control rats with the same birth date. The difference in
enzyme activity for the two studies may have been due to
a modification of formaldehyde detection methods (Venkatesan,
et al. 1968) . In these studies, no value below ten percent
was considered as significant activity. Arcos, et al. (1976)
noted that demethylation is a requirement for carcinogenesis
by DMN, and thus it is possible that acenaphthene may slightly
inhibit DMN carcinogenesis.
Buu-Hoi and Hien-Do-Phouc (1969) investigated the effect
of acenaphthene and other polycyclic aromatic hydrocarbons
(PAHs) on the activity of zoxazolamine hydroxylase. Male
Wistar rats were injected intraperitoneally with 20 mg/kg
acenaphthene in corn oil, followed one week later by 90
mg/kg zoxazolamine. The mean paralysis time of treated
rats was found to be significantly greater (p<0.01) than
that of vehicle-injected animals. The authors interpreted
these results as an indication that acenaphthene retards
the detoxification of zoxazolamine, which ordinarily proceeds
via hydroxylation.
Teratogenicity
No information was found concerning the teratogenicity
of acenaphthene.
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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 mutagenicity was observed in any of the pro-
cedures used. Clark (1953a) studied the effect of acenaph-
thene on the recombination rate of two auxotrophic Bacterium
coli (E. coli) strains. Acenaphthene was found to have
no appreciable effect upon the recombination rate of either
strain, as indicated by the low level of prototroph induc-
tion. 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 acenaphthene administered was
not specified. In a later study, Clark (1953b) tested acenaph-
thene for mutagenicity by exposing Micrococcus pyogenes
var. aureus strain FDA209 to a saturated solution of ace-
naphthene in a water-based nutrient broth without a metabolic
activation system. When induction of mutants resistant
to penicillin or streptomycin was assessed, acenaphthene
did not demonstrate any mutagenic effects.
Two mutagenicity studies performed using Salmonella
typhimurium 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_;_ typhimur ium TA98. No revertants were
observed with or without rat liver activation. Gibson,
et al. (1978) exposed1^ typhimur ium strains to 200 to 2000
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jjg of acenaphthene dissolved in dimethylsulfoxide after
first irradiating the acenaphthene samples with ®^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 assess-
ment 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 Halonen, 1968)
examined the binding of acenaphthene to a variety of biologi-
cally important compounds as part of an unsuccessful attempt
to correlate the nucleoside-binding activity of various
chemicals with their carcinogenic potential. Acenaphthene
showed significant binding constants for caffeine and ribo-
flavin, 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 size and DNA content, are asso-
ciated with disruption of the spindle mechanism during mitosis
and the resulting induction of polyploidy. While there
is no known correlation between these effects and the bio-
logical impact of acenaphthene on mammalian cells, these
effects are reported in this document because they are the
only substantially investigated effects of acenaphthene.
Ten experiments examining the effect of acenaphthene
on plants and eight others involving the effects upon micro-
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organisms are discussed in the following sections. A sum-
mary of these data is presented in Table 1.
Plants: Kostoff (1938a) exposed Nicotiana longiflora
shoots to vapors from acenaphthene crystals and examined
the shoots for effects on mitosis and/or meiosis. The expo-
sure induced tetraploid and octaploid shoots, which produced
seeds of new polyploid plants. The polyploidizing effect
of acenaphthene vapor increased with increases 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
failing to arrange correctly on the equatorial plate. They
tended to spread into the cytoplasm singly or in groups,
resulting in a variable 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 meiosis was
abortive.
In the 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, maize, and rice) showed slow growth and
abnormal roots and leaf formations after four to eight days.
Legumes evidenced these effects after 6 to 12 days, while
compositae reacted in a time period midway between the other
two groups. Mitosis in these seedlings proceeded abnormally;
the spindle mechanism was inhibited and the chromosomes
C-ll
-------
TABLE 1
Summary of Polyploid and Other Mitotic Effects Induced
by Acenaphthene in Plants and Microorganisms.
ORGANISM
TREATMENT
EFFECTS NOTED
REFERENCE
n
N>
Plants:
Nicotiana shoots
Cereal, grass
legume, and
compositae seeds
irry-mazzard
arid seeds
Allium cepa L,
Allium cepa L.,
A. sativum
Allium fistulosum,
Colchicum roots
Allium root cells
Vapors
Crystals
(4-12 days)
Powder
(10 hours)
Saturated
solution
(2-5 days)
Treatment
unspecified
Crystals wrapped
in moist filter
paper (4-20 days)
Vapors
(12-96 hours)
Stable polyploidy;
abnormal, abortive
meiosis
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,
1938a, b
Kostoff, 1938b
Zhukov, 1971
D'Amato, 1949
Mookerjee, 1973
Levan, 1940
Mesquita, 1967
-------
TABLE 1 (cont.)
ORGANISM
TREATMENT
EFFECTS NOTED
REFERENCE
Binucleate pollen Vapors
Tradescantia
pollen
Tradescantia
stamen hairs
Fungi:
Basidomycetes
Basidiobolus
ranarum hyphae
Pythium
aphanidermatum
hyphae
Yeast
Vapors
Saturated solution
(2-4.5 hours)
Vapors
Vapors
(6-18 hours)
Vapors or
supersaturated
solution
(12 hours)
10-1 fc?-7
3 x 10 mol
Solution
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
Dyer, 1966
Swanson, 1940
Nebel, 1938
Hoover, 1972
Hoover and
Liberta, 1974
Seshadri and
Payak, 1970
Levan and
Sandwall, 1943
Candida scottii
0.2-1.0%
agar
Increase in cell
size, nucleus,
DNA content
Imshenetsky,
et al. 1966
-------
TABLE 1 (cont.)
ORGANISM
TREATMENT
EFFECTS NOTED
REFERENCE
Bacteria:
Mycobacter ium
rubrum
Rhizobium
Algae:
Chara globular is;
Nitella
flagelliformis
Vapors from
10-20 mg
crystals
Vapors
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
Tripathi,
1976a, b
-------
were not arranged on the equitorial 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 ten hours. Seed germination
and seedling growth were inhibited, but no polyploidal cells
were found in the plant roots.
Four investigators performed experiments with acenaph-
thene and Allium plants. When treated with saturated solu-
tions of acenaphthene in either tap or distilled water for
two to five days, Allium cepa L. demonstrated intense chromo-
some 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
(concentration unspecified) was found to retard the frequency
of division of Allium cepa and Allium sativum. Multiple
prophase was observed in A^ cepa.
Levan (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 spindle
were altered and the centromeres inactivated: this process
has been termed "c-mitosis" because a similar effect occurs
with colchicine treatment. Tetraploid and octaploid cells
were formed within 14 to 20 days, resulting in the formation
i
t
of root-tip swellings (c-tumorsi) in Allium. Mesquita (1967)
also investigated the effects oi acenaphthene on Allium
C -15
-------
root cells. He exposed A^_ cepa root tips to acenaphthene
vapor at room temperature for 12 to 96 hours. The reassem-
bling of the phragmoplast elements (small pieces of the
endoplasmic reticulum and Golgi bodies) in the equitorial
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,
Dyer (1966) exposed plant species with binucleate pollen
(such as Bellevalia romana, Tulbaghia natalensis, and Antir-
rhinum 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 mitosis 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) examined the effect of acenaphthene on mitosis
in plant hairs by treating stamen hairs of Tradescantia
with a saturated solution of acenaphthene in liquid media
for 2 and 4.5 hours. He found no polyploid cells and no
nuclei showing chromosomes in a metaphase condition.
Microorganisms: Several experiments have been performed
to investigate the effect of acenaphthene on microorganisms.
Hoover (1972) exposed 37 species of Basidiomycetes to acenaph-
thene vapors or media containing acenaphthene at unspecified
dose levels in order to examine effects on growth, pigment,
C-16
-------
mp^pjiology, nuclear division, and fruit body formation.
AsVrthe treatment time increased, changes in nuclear division
became, more, pronounced, with a concurrent decrease in the
mitotic frequency. Growth, pigment formation, differentia-
tion., and colonial and cellular morphology were affected
by acenaphthene treatment. A delay or prevention of light-
induced fruitbody formation occurred in one species; two
Species* developed greatly ehlarged fruitbodies as a result
of this treatment. The genetic stability of these phend-
typic changes was not demonstrated, however.
In a later experiment, Hoover and Liberta (1974) exposed
hyj>h|ae cultures of the fungus Basidiobolus r ana rum to acenaph-
thene vapors for 6 to 18 hours. At the end of 18 hours,
gross alterations in nuclear division were observed and
the spindle fibers were rendered unstainable. The time
required for division was significantly increased in ace-
naphthene-treated cells. The effect of acenaphthene on
fungi was also investigated by Seshadri and Payak (1970).
They exposed hyphae of Pythium aphanidermatum to acenaph-
thene vapors or to a supersaturated solution of acenaphthene
for 12 hours. The vapors proved "instrumental" in arresting
the progress of nuclear division. A marked increase in
the size and sporangia nuclei was noted, and the nuclei
showed various degrees of pyknosis and shape irregularity.
Levan and Sandwall (1943) examined the effect of vary-
1 7
ing concentrations of acenaphthene (10 to 3x10 mol solu-
tion in ethanol) on wort yeast cell cultures. Even at the
highest concentration, there was no lethality or effect
C-17
-------
on cell propagation. The authors concluded that the c-mitotic
action demonstrated by acenaphthene 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 acenaphthene
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 (pg per cell) was higher in acenaph-
thene-treated cells, although the difference between experi-
mental and control cultures decreased as the cultures aged.
Imshenetsky and Zhil'tsova (1973) attempted to produce
"polyploid-like" cells by exposing Mycobacterium rubrum
to vapors from 10 to 20 mg acenaphthene. When the vapors
were used alone for treatment, there was no increase in
the size of the cells, nor any indication of the induction
of polyploidy. When the cells were treated with water or
EDTA to increase membrane permeability, acenaphthene vapor
treatment caused elongation 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 Rhizobium
(nodule-forming pea bacteria) with acenaphthene vapors (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-18
-------
Acenaphthene has also been shown to affect mitosis
in two species of algae. Sarma and Tripathi (1976a,b) treated
Chara globular 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 chromo-
somes were seen to clump at metaphase after 120 hours.
Nine percent of the globular is cells showed complete
chromosome doubling by the end of the treatment period.
Carcinogenicity
Very little work has been done to determine whether
acenaphthene may have carcinogenic properties. Neukomm
(1974) reported negative results in a predictive test for
carcinogenicity based upon neoplastic induction in the newt
Triturus cristatus. Ten animals were injected subcutan-
eously with acenaphthene (dose and solvent not reported)
in the fleshy part of the tail along the vertebral 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 dif-
fuse tumors. Neoplastic lesions were divided into three
categories depending on the size of the lesion and assigned
a numerical coefficient accordingly: large (1.0) , interme-
diate (0.5), and limited (0.25). Calculation of a neoplastic
index by summing the coefficients of all lesions and divid-
ing by the number of observed animals gave an index for
acenaphthene of 0.0, indicating a lack of neoplastic induc-
tion in the newt.
C-19
-------
Neukomm (1974) discussed the reliability of this test
by drawing a correlation between positive index values for
a few polycyclic aromatic hydrocarbons and the carcinogenicity
of these same compounds for mouse skin. These limited com-
parisons, however, are not sufficient to establish the value
of this test for predicting carcinogenicity in mammalian
systems.
The only other carcinogenicity studies in the litera-
ture involving acenaphthene considered it as one component
of a complex mixture of PAHs. It is impossible in these
studies to sort out the relative contribution of acenaph-
thene 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-dimethyl-
benz(a)anthracene (DMBA) as the initiator. Animals were
painted once with 125 ^g DMBA on dorsal skin; three to four
weeks later the fractions were applied five times a week
for 13 months. The fraction containing acenaphthene, pyrene,
phenanthrene, and other PAHs, 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 gasoline exhaust condensates were carcinogenic in mouse
skin painting tests. This study is of interest considering
C-20
-------
a later study by Grimmer, et al. (1977) which showed that
acenaphthene was present in an unspecified concentration
in the benzene extracts of gasoline exhaust condensate.
Unfortunately, the possible contribution of acenaphthene
to the observed carcinogenicity (Hoffman and Wynder, 1962)
cannot be determined from this limited evidence.
C-21
-------
CRITERION FORMULATION
Existing Guidelines and Standards
No existing guidelines or standards were found.
Current Levels of Exposure
Virtually no information is available concerning the
prevalence or concentration of acenaphthene in the environ-
ment. Acenaphthene 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 several fossil fuel
oils. It has also been reported in wastewater from petro-
chemical, pesticide, and wood presevative industries (U.S.
EPA, 1978b) and detected in water from a river in the Netherlands
(Meijers and Van der Leer, 1976).
Special Groups at Risk
Individuals working with c,oal tar and/or its products
face a possible risk due to increased exposure to acenaph-
thene, although no data are available to estimate this risk.
Basis 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) are inadequate for
use as the basis of a criterion due to deficiencies in
the experimental designs (lack of controls, small number
of animals, etc.). Therefore, until more toxicological
data are generated, particularly teratogenic data in view
of the effects of acenaphthene on cell division, an interim
C-22
-------
criterion based upon organoleptic data is proposed. The
lowest levels eliciting human responses were reported at
0.022 to 0.22 ppm (Lillard and Powers, 1975), and thus 0.02
ppm (0.02 mg/1) is the recommended criterion.
Since the recommended criterion is based on organoleptic
effects and is not a toxicological assessment, the consumption
of fish and shellfish products will not be considered as
a route of exposure.
It must be emphasized, however, that this value is
not related to health effects and that the significance
of odor thresholds is unknown. This criterion will need
to be reviewed once more toxicological data are available.
C-23
-------
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