-------
TABLE 1-1
Manufacturers of Benzole Add 1n the United States
as of January, 1986*
Manufacturer and Location Annual Capacity
(millions of pounds)
Velslcol Chem. Corp.
Beaumont, TX 50
Chattanooga, TN 60
Kalama Chem. Inc.
Garfleld, NJ 15
Kalama, WA 140
Pfizer Inc.
Terre Haute, IN 9
TOTAL 274
'Source: SRI, 1986
0007d -3- 04/03/87
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1.4. USE DATA
According to CMR (1984), benzole acid has the following use pattern:
phenol manufacture, 54%; plastldzers, 18%; benzoyl chloride, 13%; sodium
benzoate, 8%; alkyd resins, 3%; butyl benzoate, 2%; others, 2%. This
chemical 1s also used as a food preservative and antlfungal agent, 1n
seasoning tobacco, 1n flavors, perfumes, dentrlflces, and as a standard In
analytical chemistry (Hawley, 1981).
1.5. SUMHARY
Benzole acid 1s a white solid at ambient temperatures with an odor
characteristic of benzoin or benzaldehyde. It Is soluble In most common
organic solvents but 1s only slightly soluble 1n water (Hawley, 1981). As
of January, 1986, three U.S. companies at five sites manufactured this
chemical (SRI, 1986; USITC, 1986). The reported U.S. production of benzole
add 1n 1984 was 165 million pounds and the projected demand for 1988 1s 179
million pounds (CMR, 1984). Benzole add can be manufactured by decarboxyl-
atlon of phthallc anhydride; hydrolysis of benzotrlchlorlde; oxidation of
toluene; or from benzoin resin (Hawley, 1981). Some of the uses for benzole
add 1n the United States are 1n the manufacture of phenol, plastldzers,
benzoyl chloride and alkyd resins (CMR, 1984).
0007d -4- 04/03/87
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2. ENVIRONMENTAL FATE AND TRANSPORT
2.1. WATER
The fate of benzole acid In water will depend on Us ability to undergo
microblal degradation, photochemical and chemical reactions, and to parti-
tion from the water column Into air and sediment. Of these processes, the
blodegradatlon process 1s the most thoroughly studied. The ready blodegrad-
abllHy of benzole add or Us sodium salt by pure cultures of microorgan-
isms was demonstrated by Neujahr and Varga (1970) and Banerjee et al.
(1984). WHh Trlchosporon cutaneum as pure culture, sodium benzoate showed
a 10- to 30-mlnute lag period before Initiation of degradation (Neujahr and
Varga, 1970). Benzole acid Is also readily blodegraded by mixed micro-
organisms. Phenol-adapted mixed microorganisms blodegraded the equivalent
of 6454 of the BOOT In 1.5 hours (Chambers et al., 1963). Thorn and Agg
(1975) reported that benzole acid was easily blodegraded by microorganisms
under biological sewage treatment conditions. Using adapted activated
sludge as microorganisms, 99% of benzole acid was removed 1n 5 days of
Incubation when the percent removal was based on COD (PHter, 1976). Lund
and Rodriguez (1984) reported -98% degradation of benzole acid In 20 days
with phenol-acclimated sludge. About 66% of benzole acid was completely
mineralized to carbon dioxide with activated sludge 1n 5 days (Freltag et
al., 1985). Rubin et al. (1982) and Mills and Stack (1954) reported the
blodegradabHlty of benzole acid by mixed microorganisms from sewage. At an
Initial concentration of 59 mg/i, 99.5% of benzole add mineralized 1n 7
days with sewage seed as microblal Inoculum (Rubin et al., 1982).
Although the above studies demonstrate the easy b1odegradab1lHy of
benzole add, they cannot be used for predicting the blodegradatlon half-
life of this compound 1n natural water. Lu and Metcalf (1975) studied the
0007d -5- 05/12/87
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blodegradaUon of benzole add 1n an aquatic ecosystem and concluded that
this chemical 1s easily degradable 1n natural water.. Benzole acid at
Initial concentrations of 15-18 mg/j. was blodegraded with half-lives of
1-4 days by two natural surface waters (Banerjee et al., 1984). At Initial
concentrations of 59 yg/l and 59 mg/i, >94.5% of benzoate mineralized
1n 7 days with two lake waters (Rubin et al., 1982). Evidence was also
provided that blodegradatlon at trace levels are different from blodegrada-
tlon at higher concentrations and that mineralization of benzoate Involves
little or no Incorporation of carbon Into mlcroblal cells. In other words,
mineralization of benzoate 1s not usually affected by the presence of
suspended solids and sediments 1n water (Subba-Rao and Alexander, 1982;
Subba-Rao et al., 1982).
The anaerobic blodegradatlon of benzoate was studied by several Investi-
gators. A stable methanogenlc bacterial mixture enriched from sludges or
sediments was shown to mineralize benzoate Into methane and carbon dioxide
(Horowitz et al., 1982; Sufllta et al., 1982; Grblc-Galk and Young, 1985;
Shelton and Tledje, 1984). Sleat and Robinson (1983) reported that long
adaptation times were required before methanogenesls of benzoate occurred.
Enriched methanogenlc mixtures obtained from sewage sludge almost completely
converted benzoate Into methane and carbon dioxide 1n <4 days (Grb1c-Gal1c
and Young, 1985). Similarly, sludge from municipal digesters almost
completely mineralized benzoate 1n 7 days under anaerobic conditions.
Freshwater lake sediment also anaeroblcally mineralized benzoate In 7 days
(Horowitz et al., 1982). This anaerobic process can be used for Industrial
wastewater treatment (Speece, 1983).
0007d -6- 05/12/87
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The suggested pathway for the methanogenesis of benzole add Is as
follows (Keith et al., 1978):
benzole acid -» 1-cyclohexene-l-carboxyllc add -» cyclohexane
carboxyllc add -» heptanolc add -» valeric add -» acetic
acid f propanolc add -» methane + carbon dioxide
Few studies are available on the fate of benzole add with respect to
chemical reactions In aquatic media or any other media. Benzole add does
not contain functional groups susceptible to hydrolysis. Therefore, this
compound 1s unlikely to hydrolyze In most aquatic media. The rate constant
for the oxidation of benzole add with hydroxyl radicals 1s 4.3xl09 M-sec
(Dorfman and Adams, 1973). Given this rate constant and a value of
3xlO~17 M as the concentration of OH radicals 1n eutrophlc water (Mill and
Mabey, 1985), the half-life for this reaction Is estimated to be >60 days.
Therefore, this reaction -Is not expected to be significant 1n water. Suzuki
et al. (1982) reported detection of mutagenlc compounds after Irradiation of
water containing nitrite and benzole add with long wavelength light;
however, the significance of this reaction In natural water cannot be
evaluated since the nitrite concentration used 1n this experiment was much
higher (>16 ppm) than concentrations present In most natural waters.
Draper and Crosby (1983) studied the possibility of photochemical oxida-
tion of benzole acid by Irradiating 1.0 mmol/i of the compound 1n water 1n
July sunlight (Davis, CA) for 5 hours; no oxidation products were detected.
Freltag et al. (1985), on the other hand. Irradiated this compound adsorbed
to silica gel for 17 hours with light of wavelength >290 nm and observed
-10% mineralization of the compound to carbon dioxide. Although the
Intensity of light used by these Investigators was greater than sunlight, 1t
appears that some photodegradatlon of adsorbed benzole add Is possible
under sunlight. . The UV absorption spectra of benzole add 1n both methanol
0007d -7- 05/12/87
-------
and potassium hydroxide show "little absorption at wavelengths >290 nm
(Sadtler, n.d.J. Based on absorption spectra and the Investigation of
Draper and Crosby (1983) H has been concluded that this compound will not
undergo significant photodegradatlon 1n natural waters.
Pertinent data regarding the rate of volatilization of benzole add from
water could not be located In the available literature as cited In Appendix
A. Based on the estimated Henry's Law constant of 2xlO~8 atm-m3/mol and
the volatility characteristics associated with various values of Henry's Law
constant (Lyman et al., 1982), 1t has been concluded that significant
volatilization of benzole acid from water 1s unlikely. Data on the removal
of benzole add from the aqueous phase by sorptlon onto suspended solids and
sediments could not be located In the available literature as cited 1n
Appendix A. Using the equation, log K = -0.55 log S * 3.54 (Lyman et
al., 1982) and the value of solubility given In Section 1.2., the K for
benzole add Is estimated as 45. Based on this value and Us K capacity
(Section 2.3.), It Is concluded that H 1s unlikely that significant amounts
of this compound may be removed through sorptlon by suspended solids and
sediments.
The bloconcentratlon factor of benzole add In the fish, golden Ide,
Leudscus j_dus melanotus. was reported to be <10 (Freltag et al., 1985).
Therefore, this compound will not bloconcentrate significantly In aquatic
organisms.
2.2. AIR
Information on the fate of benzole add In air 1s limited. Given the
assumption that the first-order reaction rate constant should be about the
same 1n the gas and liquid phases (Guesten et al., 1981), the rate constant
for OH radical reaction with benzole add In the gas phase will be
0007d -8- 04/03/87
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7.14x10 12 cm3/molecule-sec (Dorfman and Adams, 1973). Using this rate
constant and a value of 106 radicals/cm3 for the concentration of OH
radicals In the atmosphere, the half-life of benzole add 1n the atmosphere
Is -1 day. Based on photolysis data 1n aquatic media, 1t has been concluded
that benzole add 1s not likely to undergo significant photolysis In the
atmosphere. Benzole add 1s expected to behave like other adds and react
with atmospheric bases, such as ammonia and amines to form salts; however,
Information about such reactions could not be located 1n the available
literature as dted 1n Appendix A.
Since the vapor pressure of benzole add at ambient temperatures 1s
>10~4 mm Hg (see Section 1.2.), this compound 1s expected to exist predom-
inantly 1n the vapor phase 1n the atmosphere (E1senre1ch et a!., 1981).
Based on Us aqueous solubility (see Section 1.2.), this compound 1s
expected to be removed partially from the atmosphere by wet deposition. It
has been detected In rainwater and 1n snow 1n three locations 1n Norway
(Lunde et al., 1977).
2.3. SOIL
The b1odegradab1Uty of benzole add by microorganisms Isolated from
soil was studied by several authors. A species of Pseudomonas bacterium
Isolated from soil was shown to degrade this compound (Kllpl et al., 1980).
For example, Tabak et al. (1964) reported oxidation equivalent to -41% of
BOOT with phenol-adapted mixed microorganisms In 3 hours, and Alexander and
Lustlgman (1966) reported almost complete degradation of benzole add In 1
day with mixed microorganisms derived from soil. In natural soils, Mailer
(1978) reported that the blodegradatlon of benzole add required a 0- to
1-day lag period. Ward (1985) reported that benzole add Is rapidly bio-
degraded 1n surface soils by microorganisms under aerobic conditions and In
0007d -9- 05/12/87
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subsurface soils under both aerobic and anaerobic conditions. In a sub-
surface sand, the half-life for aerobic biodegradation was -7 hours, and for
anaerobic biodegradation was -17 hours (Ward, 1985). The capability of
heterotrophic bacteria in groundwaters to biodegrade benzole add was
reported by Ventullo and Larson (1985). In a groun'dwater sample from
Canada, the authors estimated that the complete mineralization half-life for
benzole add was ~3 days.
Pertinent data regarding the fate of benzole add 1n soils as a result
of chemical reactions or Interaction with sunlight could not be located 1n
the available literature as cited 1n Appendix A; however, based on Us
predicted fate In water, H 1s unlikely that either photolysis or hydrolysis
will be significant 1n soils.
WHh montmorlllonlte clay, Bailey and White (1970) observed no adsorp-
tion of benzole acid under addle or neutral conditions. Loekke (1984) also
reported no adsorption of this compound by sandy clay loam or sandy loam
soils, but a sandy clay soil showed adsorption with a K value of 230.
Therefore, It Is concluded that benzole acid will show medium to very high
mobility 1n most soils.
Based on Us estimated volatility from aquatic media, 1t 1s predicted
that benzole add will not significantly volatilize from soils.
2.4. SUMMARY
Few studies are available on the fate of benzole acid as a result of
chemical reactions. Since 1t does not contain any hydrolyzable functional
groups, 1t has been predicted that hydrolysis will not be a significant
process 1n aquatic media. Reaction with OH radicals In aquatic media 1s
also not likely to be significant (Dorfman and Adams, 1973). From the UV
absorption spectrum of this compound at wavelength >290 nm (Sadtler, n.d.)
0007d -10- 04/03/87
-------
and the study of Draper and Crosby (1983), H has been concluded that this
compound win not undergo significant photodegradatlon In natural waters.
Both pure culture and mixed microorganisms studies Indicate that this
compound 1s readily biodegradable (Banerjee et a!., 1984; Freltag et al.f
1985; Rubin et al., 1982). The blodegradatlon half-life of benzole add 1n
most natural waters 1s expected to be 1-4 days (Banerjee et al., 1984; Rubin
et al., 1982). It has also been shown that the mineralization rate of
benzoate Is not usually affected by the presence of suspended solids or
sediments In water (Subba-Rao and Alexander, 1982; Subba-Rao et al., 1982).
Benzole add 1s also susceptible to anaerobic blodegradatlon, although such
blodegradatlon may require a longer acclimation period (Horowitz et al.,
1982; SuflUa et al., 1982; Grb1c-Gal1c and Young, 1985). The anaerobic
process has practical significance 1n that It can be used for Industrial
wastewater treatment (Speece, 1983) and H Indicates that blodegradatlon of
benzole add may occur In bottom sediments.
Based on an estimated Henry's Law constant, H has been concluded that
this compound will not volatilize significantly from water. Neither adsorp-
tion nor bloconcentratlon 1n aquatic organisms are expected to be signifi-
cant for benzole add (Freltag et.al., 1985; Loekke, 1984; Bailey and White,
1970).
Based on Us expected photolytlc behavior 1n water, 1t has been con-
cluded that significant photolysis 1n air Is unlikely. Based on the rate
constant In water, the estimated half-life for Us reaction with OH radicals
1n the atmosphere 1s -1 day. No Information about the reaction of benzole
add with atmospheric bases was available In the literature. Removal of
atmospheric benzole acid by rainwater and snow has been observed (Lunde et
al., 1977).
0007d -11- 04/03/87
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On the basis of Us expected reactivity In water, It 1s concluded that
neither hydrolysis nor photolysis will be significant In soils. Isolated
microorganism, mixed soil microorganism and soil blodegradatlon studies show
that benzole add Is easily biodegradable 1n soils (Kllpl et al., 1980;
Tabak et al..- 1964; Alexander and Lustlgman, 1966; Ward, 1985). In a
subsurface sand, the half-life for blodegradatlon was 7 hours under aerobic
conditions (Ward, 1985). This compound 1s also susceptible to anaerobic
blodegradatlon. The half-life for anaerobic blodegradatlon 1n a subsurface
soil was 17 hours (Ward, 1985). Based on Us estimated volatility 1n water,
1t 1s predicted that benzole add will not volatilize significantly from
soils. Benzole acid will show medium to high mobility In soils (Bailey and
White, 1970; Loekke, 1984). Therefore, H 1s likely to leach from most
soils to groundwater.
0007d -12- 05/12/87
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3. EXPOSURE
3.1. AIR
Benzole add was detected 1n the process exhaust from a phthallc
anhydride manufacturing plant at concentrations of 5-40 ppm (v/v); however,
application of pollution abatement devices 1s expected to reduce the
pollutant level (Fawcett, 1970). Benzole acid at a concentration of 0.164
ppb was also detected 1n the exhaust of gasoline-powered automobiles.
Although a new engine oil showed no benzole add, used oil from a similar
automobile engine contained 45.3 ppb of the compound (Kawamura et a!.,
1985). Similarly, other authors reported the detection of benzole acid 1n
the exhaust of dlesel-powered vehicles (Hampton et al., 1982). Kawamura et
al. (1985) reported benzole add at a mean concentration of 0.010 ppb 1n the
Los Angeles atmosphere.
3.2. WATER
Benzole acid was detected 1n Industrial and municipal effluents,
leachates from waste disposal sites, raw surface water and drinking water.
The compound was tentatively Identified In the primary and final effluents
from POTWs (Ellis et al., 1982; U.S. EPA, 1975) and 1n the effluents from a
pulp mill plant (Undstrom and Osterberg, 1986). Frands et al. (1980)
detected benzole add at a maximum concentration of 6.7 ppm from the
leachates of two low level radioactive disposal sites 1n Maxey Flats, KY,
and West Valley, NY. The maximum concentration of benzole acid detected 1n
leachates from two landfill sites 1n Ontario, Canada was >1 ppm (Relnhard et
al., 1984). This compound was detected 1n the groundwater at concentrations
as high as 0.86 ppm from an underground coal gasification site 1n north-
eastern Wyoming (Stuermer et al., 1982), and at 27.5 ppm from a wood-
preserving facility 1n Pensacola, FL (GoerlHz et al., 1985). Stepan et al.
0007d -13- 04/03/87
-------
(1981) qualitatively detected benzole acid In the groundwater from a
chemical waste disposal site near Melbourne, Australia. Benzole add was
also present in surface waters in England (Fielding et a!., 1981} and 1n
Norway (Schou et al., 1981). Benzole add at a concentration of 15 ppb was
reported In the drinking water from Ottumwa, IA (U.S. EPA, 1975; Kopfler et
al., 1977).
3.3. FOOD
Benzole add 1s present naturally 1n many foodstuffs. FEMA (1984)
reported that levels ranged from a minimum of 0.239 ppm 1n apple wine to a
maximum of 40 ppm In apple essence. The greater contribution to the human
diet, however, results from the addition of benzole acid and sodium benzoate
to food as an antimicrobial preservative (Chapter 7). FASEB (1973) reported
results of a comprehensive survey by the Subcommittee on Review of the GRAS
List (1972) from which possible dally dietary Intakes of benzole acid and
sodium benzoate were estimated (Table 3-1). The report acknowledged that
these data probably represent a gross exaggeration of actual dietary
Intakes. Based on data regarding the amounts of benzole add and sodium
benzoate produced for addition to food, FASEB (1973) estimated dally per
capita dietary Intakes of benzole add of 0.9 mg and of sodium benzoate of
34 mg.
3.4. SUMMARY
Benzole add was reported to be present 1n the exhaust from gasoline and
dlesel-powered vehicles (Hampton et al., 1982; Kawamura et al., 1985). A
mean concentration of 0.010 ppb was detected 1n the Los Angeles atmosphere
(Kawamura et al., 1985). Human exposure to benzole add 1n the United
States from Inhalation of air cannot be estimated until more monitoring data
are available. Benzole acid was detected In Industrial and municipal
0007d -14- 04/03/87
-------
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Ing water (Ellis et a!., 1982; Lindstrom and Osterberg, 1986; Francis et
al., 1980; Reinhard et al., 1984; Stuermer et al., 1982; Goerlitz et a!.,
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concentration of 15 ppb was reported in the drinking water from Ottumwa, IA
(U.S. EPA, 1975). Human exposure to this compound from ingestion of drink-
Ing water in the United States cannot be estimated until more drinking water
monitoring data are available. Benzole acid 1s present naturally 1n some
foods. Benzole add and sodium benzoate are also added to food as preserva-
tives. Based on production data, estimated per capita daily Intakes are 0.9
mg for benzole add and 34 mg for sodium benzoate (FASEB, 1973).
0007d -16- 04/03/87
-------
4. AQUATIC TOXICITY
4.1. ACUTE TOXICITY
Data concerning acute toxldty of benzole add to aquatic organisms are
presented In Table 4-1. Interpretation of these data are complicated by the
fact that the form of the add (free add, neutralized add or salt) used In
the studies was not always stated explicitly. According to Doudoroff and
Katz (1950), the toxldty of benzole add 1s due primarily to the undlsso-
dated add 1n solution and not to pH depression.
All of the available data apply to freshwater species. The lowest
reported toxic concentration for freshwater fishes was 180 mg/i, a 96-hour
LC__ for mosquHoflsh, Gambusla afflnls (Wallen et a!., 1957). Among
Invertebrates, the lowest reported toxic concentration was 31 mg/l, a
threshold for Inhibition of cell multiplication 1n the protozoan, Uronema
parduczl (BMngmann and Kuehn, 1981). Additional data were provided by
Ewell et al. (1986) who Indicated that 96-hour LC5Q values for the follow-
ing species were all >100 mg/l: plllbug, Asellus Intermedus; water flea,
Daphnla magna; flatworm, Duqesla tlqrlna; s1desw1mmer, Gammarus fasdatus;
snail, Hellsoma trlvolvls; segmented worm, LumbMculus varlegatus; and
fathead minnow, Plmephales promelas.
4.2. CHRONIC EFFECTS
Pertinent data regarding chronic toxldty of benzole add to aquatic
organisms could not be located 1n the available literature as cited 1n
Appendix A.
4.3. PLANT EFFECTS
The available data concerning toxldty of benzole add to aquatic plants
are presented In Table 4-1. The most sensitive of three algal species
tested was the blue-green alga, M1crocyst1s aeruqlnosa, with a threshold of
55 mg/l for Inhibition of cell multiplication (Brlngmann and Kuehn, 1978).
0007d -17- 05/12/87
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4.4. SUMMARY
The data concerning aquatic toxlclty of benzole acid were limited. The
lowest reported toxic concentration was 31 mg/j., a threshold for Inhibi-
tion of cell multiplication 1n the protozoan, Uronema parduczl (Brlngmann
and Kuehn, 1981).
0007d -20- 04/03/87
-------
5. PHARMACOKINETICS
Benzole add Is used as a food preservative (FEMA, 1984) and has limited
use as a pharmaceutical [e.g., 1n the treatment of hyperammonemla 1n Infants
(Green et al., 1983)]. Consequently, the pharmacoklnetlcs of this compound
has been Investigated both In humans and animals. Table 5-1 summarizes
several pharmacoklnetlc studies of benzole add 1n several spedes.
5.1. ABSORPTION
Informatics, Inc. (1972) concluded that the gastrointestinal absorption
of benzole acid In humans Is rapid and complete. Urinary excretion by
\
humans of 95-99% of oral doses confirms this conclusion (see Table 5-1)
(Bridges et al., 1970; R11hmak1, 1979). Urinary excretion In subhuman
primates ranged from 33-75.5X of an oral dose (Bridges et al., 1970; Hall
and James, 1980). Virtually complete gastrointestinal absorption has also
been shown for rats (Hall and James, 1980; Jones, 1982; Bridges et al.,
1970), hamsters and dogs (Bridges et al., 1970). For spedes 1n which
urinary recovery amounted to ~30-60% of the dose, 1t Is not clear 1f the
excretion data reflect saturation of absorption, urinary excretion or
blotransformatlon mechanisms; excretion by an alternate route; or tissue
retention resulting 1n less material available for excretion.
Most of the data 1n Table 5-1 reflect excretion within 24 hours. The
recovery of the equivalent of virtually the entire dose of benzole add from
the urine within 24 hours Indicates that the material was rapidly absorbed
from the gastrointestinal tract. R11hmak1 (1979) noted that urinary excre-
tion of hlppurlc add, the major metabolite of benzole add 1n humans, was
rapid, with a rate constant of 1.0 hour'1 within 4-5 hours of treatment.
In rats, excretion was virtually complete within 6 hours of treatment (Hall
and James, 1980).
0007d -21- 05/12/87
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Since benzole add 1s a weak organic acid with a pK of 4.2 (Serjeant
a
and Dempsey, 1979), Us gastrointestinal absorption would be expected to
decrease as pH Increased above 4.2 and to Increase as pH dropped below 4.2.
Using an _1_n situ preparation of rat jejenum, Hoegerle and W1nne (1983)
demonstrated that absorption decreased as the pH of the perfuslon solution
was raised from 4-10.5. The absorption curve, however, was considerably
flatter than that predicted by the unmodified pH-partH1on theory, and more
nearly reflected pH as measured 1n the "unstirred" layer of perfuslon
solution 1n contact with the Jejunal v1H1 rather than pH as measured In the
lumen. In an earlier experiment, Ochsenfahrt and Wlnne (1974) showed that
altering the tonlcHy of the perfusing solution 1n a preparation of rat
Jejunum Influenced the rate of absorption. A positive net flux of water
from the Intestinal lumen to the circulation (hypotonlc perfusing solution)
Increased the appearance of l4C-benzo1c acid-associated radioactivity In
the blood by up to 47/4. A negative net flux of water (movement from
circulation to gut lumen) created by perfuslon with a hypertonlc solution
retarded uptake of l4C-benzo1c acid-associated radioactivity by up to 28%.
Data were not located regarding the pulmonary absorption of benzole
add. Huang et al. (1985), however, demonstrated that both the Ionized and
un-1on1zed forms of benzole add are absorbed from solution by the nasal
epithelium of rats. Using rats surgically altered to permit perfuslon of
the nasal cavity, these Investigators circulated 20 ml of 0.0015 M
solutions of sodium benzoate at pH 2.4-7.2 at a rate of 2 ma/m1nute for 60
minutes. Absorption of benzole add was determined to be 44% at pH 2.5 and
only 13% at pH 7.19. These data suggest that absorption of benzole add
through the respiratory epithelium may be expected and that the rate of
0007d -25- 05/12/87
-------
absorption may depend upon the physical form of the material 1n contact with
the epithelium and upon conditions that Influence pH at the epithelial
surface.
The dermal absorption of benzole acid has been studied using guinea pigs
and j_n vitro skin preparations from humans and rats. In the guinea pig
study, an estimated 31.4% of a 4 yg/cm2 dose 1n acetone was absorbed 1n
a 24-hour period (Andersen et a!., 1980). 14C-Labeled benzole add was
used and radioactivity was determined 1n urine collected for 5 days. The
guinea pig system was "calibrated" by measuring urinary recovery of radio-
activity following 1ntraper1toneal Injection of the labeled compound. In
the Ui vitro experiment (Bronaugh and Stewart, 1985), 24-hour absorption of
benzole add applied at 5 mg/cm2 1n a commonly-used cosmetic lotion was
27.5% 1n human skin and 19.9% 1n rat skin. Abrading -6.4% of the skin
surface by scratching through the epidermis with a hypodermic needle
Increased percutaneous absorption -2-fold.
5.2. DISTRIBUTION
Quantitative data regarding the tissue distribution of benzole acid and
Its metabolites could not be located 1n the available literature as dted 1n
Appendix A. The rate and extent of benzole add elimination, however,
suggest that tissue retention 1s not highly significant 1n the pharmaco-
klnetlcs and toxldty of the compound (FASEB, 1973).
5.3. METABOLISM
The results of the pharmacok1net1c studies summarized 1n Table 5-1
Indicate that hlppurlc add Is the predominant metabolite 1n mammals and
that benzoyl glucuronlde 1s secondary. In human experiments (Bridges et
a!., 1970; R11hmak1, 1979; Amsel and Levy, 1969) and In many rat (Bridges et
a!., 1970; Hall and James, 1980; Jones, 1982) and hamster (Bridges et al.,
0007d -26- 05/12/87
-------
1970) experiments, >90% of the dose was recovered as hlppurlc acid within 24
hours. Hlppurlc add Is formed by conjugation of benzole add with the
amlno add glydne; benzoyl glucuronlde results from conjugation with glucu-
ronlc add. In the experiments summarized 1n Table 5-1, benzoyl glucuronlde
accounted for 0-22% of the dose.
Several experiments In humans and animals show that the conversion of
benzole add to hlppurlc add Is a rate saturable process dependent upon the
availability of glyclne. Amsel and Levy (1969) estimated a maximum rate of
formation of hlppurlc add In humans of 1730 mg/hour with an estimated rate
constant of 1.2 hour"1. Increasing the dose to 5.0 g (-55.6 mg/kg)
Increased the excretion of benzoyl glucuronlde from 1.8 to 3.2-3.4% of the
dose, but the maximum rate of hlppurlc add formation only Increased to
2090-2100 mg/hour. When glydne (5 g, 1 hour before the sodium benzoate and
2 g each hour thereafter) was given with the larger dose of sodium benzoate,
the maximum rate .of hlppurlc add formation Increased to 4050 mg/hour and
the rate constant to 1.4 hour"1; the excretion of benzoyl glucuronlde
dropped to the equivalent of 0.6% of the dose.
R11hmak1 (1979) (see Table 5-1) estimated a maximum rate of hlppurlc
acid synthesis of 180-198 ymol/mlnute (-1320-1450 mg/hour) at 2 hours 1n a
human. This rate remained constant for an additional 3 hours, and a rate
constant of 1.0 hours"1 was estimated. The excretion rate then fell
rapidly until -8 hours after treatment. R11hmak1 (1979) suggested that the
3-hour plateau 1n the excretion rate Indicated that the pathway for metabo-
lism of benzole acid to hlppurlc add had become saturated.
In an early experiment, Quick (1931) administered oral doses of 1, 2, 3
or 5 g benzole add with or without the simultaneous administration of
glydne to a 52 kg man and measured urinary hlppurlc add over a 3- to
0007d -27- 05/12/87
-------
6-hour post-treatment period. At every dosage level the simultaneous oral
administration of glydne resulted 1n more rapid excretion of hlppuMc add,
attributed to more rapid conversion of benzole to hlppurlc add rather than
to an alteration of the rate of renal clearance.
Data from studies of orally-treated adult and 4- to 7-day-old marmosets
(Hall and James, 1980) suggest that a dose-dependent saturation of the con-
version of benzole add to hlppurlc add occurred, resulting 1n a propor-
tionate decrease 1n hlppurlc add excretion and Increase 1n benzoyl glucuro-
nlde and unchanged benzole add excretion. No evidence of saturation In the
formation/excretion of hlppurlc add, however, was observed 1n adult rats
treated with oral doses of benzole acid ranging from 0.01-1000 mg/kg (Hall
and James, 1980; Jones, 1982). In an j_n vitro study using Isolated hepato-
cytes from Sprague-Oawley rats, however, Wendler and Tremblay (1982) noted
that conversion of benzoate to hlppurate was limited by the availability of
glydne. Adding exogenous glydne during a 1-hour Incubation accelerated
hlppurate formation by 420%.
In weanling rats that had a marasmlc-kwashlorkor condition, which was
due to consumption of a 3.45% protein diet for 5 weeks, Thabrew et al.
(1980) noted a marked decrease 1n their ability to metabolize benzole add
to hlppurlc add. These rats excreted 59.9% of the dose as hlppurlc add
and 20.3% as benzoyl glucuronlde. Normally-fed rats excreted 81.2% of a 200
mg/kg 1ntraper1toneal dose as hlppurlc add and excreted only a trace as
benzoyl glucuronlde. The Investigators suggested that the Increased Impor-
tance of glucuronlc add conjugation 1n the marasmlc-kwashlorkor rats
reflected decreased glydne available for conjugation and Increased UDP-
glucuronyl transferase activity In protein deficient rats. When marasmlc-
kwashlorkor rats were returned to the control diet, normal conversion of
benzole add to hlppurlc acid returned 1n 2 weeks.
0007d -28- 05/12/87
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As sodium benzoate 1s occasionally used therapeutlcally In newborn
Infants to treat apnea associated with prematurity and hyperammonemla
resulting from congenital defects 1n the urea cycle, Green et al. (1983)
gave therapeutic doses of 3.5 mmol/kg/day (-427 mg/kg/day) sodium benzoate
Intravenously to four hyperammonemlc Infants. Generally "more than half" of
the excreted material was hlppurate. The remainder was excreted as
unchanged parent compound. In this study the term "excreted" Included
material eliminated 1n the urine and collected by peritoneal dialysis.
Although no supportive data were presented, Edwards and Voegell (1984)
suggested that neonates have reduced ability compared with adults to con-
jugate benzole acid with glydne.
Balnes et al. (1978) and Hall and James (1980) noted that the metabolism
of benzole add was age-dependent 1n rats. Balnes et al. (1978) adminis-
tered l4C-sod1um benzoate (dose and route not specified) to adult rats and
recovered hlppuMc add 1n the urine nearly equivalent 1n amount to the
administered dose. In neonatal rats, however, -20% of urinary radlolabel
was Identified as benzoyl glucuronlde. The Investigators attributed these
results to higher levels of UOP-glucuronyl transferase and lower levels of
glyc1ne-N-acyltransferase 1n the livers of neonatal rats compared wHh adult
rats. A similar trend was observed In the metabolism of l4C-benzo1c add
by young vs. adult rats 1n the experiment by Hall and James (1980) (see
Table 5-1). Young rats (9- to 10-day-old) appeared to be less capable than
adults 1n transforming 40-100 mg/kg oral doses of benzole acid to hlppurlc
add. Reduced ability of the young to transform benzole add to hlppurlc
add was not apparent In young vs. adult marmosets (Hall and James, 1980)
(see Table 5-1). Conjugation with glucuronlc add 1s more Important 1n
marmosets than In humans or rats.
0007d -29- 04/03/87
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Although the predominant metabolite of benzole add produced by the
animals listed 1n Table 5-1 appears to be hlppurlc add, benzoyl glucuronlde
1s produced 1n small quantities by most of these animals and 1n significant
quantities by carnivores such as dogs and ferrets (Bridges et al., 1970).
Quick (1932) administered oral doses of benzole add to bilaterally nephrec-
tomlzed dogs. Upon sacrifice on the next day, hlppurlc acid was not found
1n the blood, and the Investigator concluded that the kidney was the sole
site of conversion of benzole acid to hlppurlc add 1n the dog.
Kao et al. (1978) demonstrated species differences In the metabolism of
benzole acid using Ui vitro preparations from hepatocytes and renal tubules
of male Wlstar rats, Syrian hamsters, a beagle dog and albino ferrets.
Ring-labeled l4C-benzo1c acid was used and metabolites were separated,
Identified and quantified. Liver preparations from the rats and hamsters
rapidly converted the dose of benzole add to nearly equivalent amounts of
hlppurlc add. In addition, small amounts of benzoyl glucuronlde (equiva-
lent to <5% of the Initial dose of benzole add) were produced. Liver
preparations from the dog and ferret, however, produced no hlppurlc add but
only benzoyl glucuronlde, equivalent to -8 and 11% of the Initial dose In
these two species, respectively. The kidney preparations from the rat, dog
and ferret converted nearly the entire dose of benzole add to hlppurlc
add. Only the kidney preparation from the rat produced a trace amount of
benzoyl glucuronlde. The kidney preparation of the hamster was not nearly
as efficient as that of the other species, as only ~13% of the dose was
transformed to hlppurlc add, the only metabolite produced.
The data reviewed In Table 5-1 Indicate that benzole add 1s extensively
metabolized 1n most species, although adult marmosets may excrete substan-
tial amounts of unchanged compound. The fate of the unrecovered portion of
0007d -30- 05/12/87
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the dose was not explained. Jones (1982) recovered -3% of an oral 0.01
mg/kg dose of [carboxy-l4C]-benzo1c add 1n rats as expired 14C02
within 24 hours. Decarboxylatlon may contribute substantially to the
elimination of benzole add In certain species or when conditions result 1n
saturation of other metabolic pathways. In addition, other minor metabo-
lites of benzole acid have been Identified In Yn vitro studies. Sato et al.
(1956) Incubated benzole acid with rat liver slices In an Incubation medium
containing 3SS-sulfate and Identified the sulfate conjugate of 4-hydroxy-
benzolc add. Liver mlcrosomal preparations from guinea pigs and rabbits
converted benzole add to "trace levels" of 3-hydroxybenzolc add (Daly et
al., 1968). These results suggest that oxidation, probably followed by
conjugation, plays a significant role 1n the metabolism of benzole add, at
least under certain conditions.
Other explanations for the Incomplete recovery Include Incomplete
gastrointestinal absorption or the excretion of benzole add and Its metabo-
lites through the bile or by direct secretion Into the Intestinal tract,
although Investigations 1n rats and mice (Section 5.4.) Indicate that these
are minor routes of excretion for these species.
5.4. EXCRETION
Data regarding the urinary excretion of benzole add and Us metabolites
are presented 1n Table 5-1. For humans, rats, hamsters and dogs, excretion
appears to occur rapidly and nearly completely through the urine. Urinary
excretion removes the metabolites hlppuMc add and benzoyl glucuronlde. In
contrast, little unchanged benzole add 1s excreted by the kidney [with the
possible exception of adult marmosets (Hall and James, 1980)]. In monkeys,
pigs, rabbits, mice, guinea pigs, cats and ferrets, urinary excretion
0007d -31- 05/12/87
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appears to account for a substantially smaller proportion of an oral dose,
although considerable Interlaboratory variation 1s noted. Route of admin-
istration appears to have no Impact on urinary excretion.
Jones (1982) determined that -3% of an oral dose of 0.01 mg/kg carboxyl
l4C-benzo1c acid was expired as 14CO_ 1n treated rats 1n 24 hours.
The respiratory tract may be a more Important route of excretion 1n other
species or with larger doses or 1n the presence of other conditions that
saturate conjugation with glydne to form hlppurlc add.
Following IntraperVtoneal administration of a 10 mg/kg dose of [car-
boxy-l4C]-benzo1c add as the sodium salt to rats, hamsters, guinea pigs,
rabbits, sheep, cats and ferrets, Huckle et al. (1981) recovered <5% of the
dose of radioactivity In the feces by 72 hours and concluded that fecal
excretion was not Important In the elimination of benzole add and Us
metabolites. In ddN mice given a 100 mg/kg 1ntraper1toneal dose of radio-
labeled benzole add (position of label not specified), followed by collec-
tion of urine, bile and feces for up to three 24-hour periods, urinary
excretion of radlolabel accounted for 100.9*1.5% of the dose (Kato, 1972).
In female Wlstar rats given a 410 ymol/kg (50 mg/kg) dose by 1ntraper1to-
neal Injection (H1rom et al., 1976) or In Donryu rats given an oral 50 mg/kg
dose (Kato, 1972) of radlolabeled benzole add, biliary excretion accounted
for 1 and 2.5% of the dose, respectively. Biliary and Intestinal excretion
may be more Important 1n species 1n which urinary recovery accounted for
smaller portions of an oral or parenteral dose.
5.5. SUMMARY
Gastrointestinal absorption of benzole acid appears to be virtually
complete and fairly rapid 1n humans (Bridges et al., 1970; R11hmak1, 1979;
Amsel and Levy, 1969), rats (Bridges et al., 1970; Hall and James, 1980;
0007d -32- 05/12/87
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Jones, 1982), hamsters and dogs (Bridges et al., 1970); urinary excretion
products account for >90% of the dose within 24 hours of treatment. I_n_ situ
studies 1n rats Indicate that Increasing Intestinal pH above 4.2 decreases
the rate of absorption (Hoegerle and W1nne, 1983) and Increasing or decreas-
ing the net flux of water from the Intestine by altering the tonlcHy of the
perfuslon solution (Ochsenfahrt and W1nne, 1974) Increases or decreases,
respectively, the uptake rate of benzole acid. Data were not located
regarding Inhalation absorption; however, 1n an j_n sjtu nasal cavity
perfuslon study 1n rats, absorption of benzole add occurred and the rate
depended on the pH of the perfusing solution (Huang et al., 1985).
The dermal absorption of 31.4% of a 4 yg/cm2 dose has been demon-
strated 1n guinea pigs (Andersen et al., 1980). WHh In vitro preparations
of human and rat skin, Bronaugh and Stewart (1985) showed that scarification
Increased percutaneous absorption ~2-fold.
Data were not located regarding the distribution or retention of benzole
add or Us metabolites, but the rapidity and extent of benzole add elimi-
nation suggest that retention 1s probably not Important 1n the pharmaco-
klnetlcs and toxlclty of the compound (FASEB, 1973).
In humans and common laboratory spedes (Bridges et al., 1970; RUhmakl,
1979; Amsel and Levy, 1969; Hall and James, 1980; Jones, 1982; Huckle et
al., 1981; Thabrew et al., 1980) hlppurlc add, formed by conjugation with
the amlno acid glydne, 1s the predominant metabolite (up to 100% of the
dose) and benzoyl glucuronlde, formed by conjugation with glucuronlc add,
Is a lesser metabolite (0-22% of the dose). The rate and extent of conver-
sion of benzole add to hlppurlc add 1s dependent upon the availability of
glydne, and can be Increased by the administration of exogenous glyclne
(Quick, 1931; R11hmak1, 1979; Amsel and Levy, 1969). The proportion
G007d -33- 05/12/87
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excreted as hlppuric acid by humans and rats Is reduced 1n the very young
(Green et al., 1983; Edwards and Voegell, 1984; Balnes et al., 1978; Hall
and James, 1980). The liver and kidney appear to be the major sites of
conversion to hlppuMc acid and benzoyl glucuronlde, but there are marked
species differences 1n the rate, extent and products of metabolism at each
site (Kao et al., 1978). C0? resulting from decarboxylatlon Is a minor
metabolite 1n rats (Jones, 1982), but Its Importance has not been Investi-
gated In other species. Ln vitro studies suggest that hydroxylatlon of the
benzene ring, probably followed by conjugation, may also occur (Sato et al.,
1956; Daly et al., 1968).
For humans, rats and dogs excretion appears to occur rapidly and nearly
completely through the urine (Bridges et al., 1970; R11hmak1, 1979, Amsel
and Levy, 1969; Hall and James, 1980). Renal excretion of the metabolites
hlppuMc add and benzoyl glucuronlde Is rapid. In rats, -3% of an oral
dose was expired as COp (Jones, 1982). This mode of elimination may be
more significant In other species or 1f preferential routes of elimination
become saturated. Biliary and Intestinal excretion has not been adequately
studied 1n those species 1n which urinary excretion accounted for a smaller
portion of the dose. In many species, fecal excretion of radioactivity
following an 1ntraper1toneal dose of [carboxy-l4C]-benzo1c add Is <5%
(Huckle et al.. 1981).
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6. EFFECTS
6.1. SYSTEMIC TOXICITY
6.1.1. Inhalation Exposures.
6.1.1.1. SUBCHRONIC IRDC (1981) reported a 4-week Inhalation study
with technical grade benzole acid In young Sprague-Dawley rats. Groups of
10 rats/sex were exposed to target concentrations of 0, 0.02, 0.2 or 2.0
mg/i, 6 hours/day, 5 days/week. Dry flaked material was used to generate
a dust aerosol with an equivalent aerodynamic diameter of 4.7 y. Actual
concentrations were measured to be 0, 0.025, 0.25 and 1.2 mg/J. (0, 25, 250
and 1200 mg/m3). Parameters of toxldty evaluated Included clinical signs
and mortality, body weight gain, hematology, blood chemistry focused on
liver function and damage, gross necropsy and organ weights, extensive
histology of controls and high-dose rats, and hlstologlc examination of the
lungs of low- and middle-dose rats.
A reddish nasal discharge was consistently observed 1n middle- and
high-dose rats, Indicating upper respiratory Irritation. One male and one
female 1n the high-dose group died during exposure. A statistically
(p<0.01) and biologically significant decrease In rate of body weight gain
occurred In both sexes In the high-dose groups. The only treatment-related
hematologlc effect was a reduction 1n platelet count 1n both sexes (p<0.01)
1n the high-dose group. Sporadic changes 1n blood chemistry were not
attributed to treatment, and there were no gross pathologic effects attrib-
uted to treatment. A decrease 1n both absolute organ weights and organ-to-
brain weight ratios was observed 1n the livers of high-dose males and the
kidneys, trachea and lungs of high-dose females. Reduced absolute kidney
weight also occurred In middle-dose females. H1stopatholog1c alterations
were restricted to the lungs and consisted of Interstitial flbrosls and
0007d -35- Ob/12/87
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Interstitial Inflammatory cell Infiltration. A dose-related Increased
Incidence and Intensity of these Inflammation effects were noted 1n all
treated groups, which was attributed to the persistent Irritating effect of
the test material.
6.1.1.2. CHRONIC -- Pertinent data regarding the chronic Inhalation
toxldty of benzole add could not be located In the available literature as
cited 1n Appendix A.
6.1.2. Oral Exposures.
6.1.2.1. SUBCHRONIC Subchronlc Investigations with both benzole
add and sodium benzoate have been conducted 1n several laboratory species
and humans. Shtenberg and Ignat'ev (1970) administered dally gavage doses
of benzole acid (reagent grade, vehicle not reported) at 80 mg/kg to a group
of 50 male and 50 female white mice for 3 months. Controls were maintained
but their numbers and treatment were not described. The mice were observed
for general condition and behavior, survival, food and water Intake, body
weight gain and, at the end of the 3-month exposure period, response to
stresses such as hunger and poisoning with carbon tetrachlorlde. In spite
of nearly equivalent food Intake, male mice gained only 63% and females 71%
as much as their respective controls. Treated mice had greater mortality
than controls when stressed with carbon tetrachlorlde poisoning.
Krels et al. (1967) fed diets containing 3% benzole add to groups of
5-15 male Royal Wlstar rats for <5 days. Effects noted by day 5 Included
weight loss, a number of CNS signs and mortality of -1/2 of the rats. The
most significant lesions were located 1n the brain and Included necrosis of
the gangHonlc cells of several different regions. Some of the rats that
survived the 5-day dietary exposure were maintained on control diets for an
additional 19-30 days, at which time they were sacrificed. The lesions that
0007d -36- 05/12/87
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were observed In the rats that died were also observed 1n surviving rats
after this recovery period. In addition, groups of 5-10 rats were main-
tained on a diet containing 1.1% benzole acid for 7, 14 or 35 days. Reduced
growth rate and Impaired food efficiency were evident, but there were no
gross or microscopic lesions In the heart, liver, kidneys or brain.
Behavioral signs of CNS Impairment, often accompanied by death were also
observed 1n cats given 1-4 doses of 300-890 mg/kg/day benzole add 1n meat
(Bedford and Clarke, 1972). Clinical observation and examination of c!1n1-
copathologlc parameters of liver and kidney function revealed no effects
among four cats treated with 130-160 mg/kg/day for 23 days.
In humans, a single oral 10 g dose of benzole add had no effect on body
temperature, pulse, respiration or other clinical parameters (Gerlach,
1909). Similarly, no externally visible effects were observed following 44
consecutive days at 0.5 or 1.0 g/day or after 82 doses 1n 86 days or 88
doses In 92 days af 1 g/day. After 6 days of treatment, there were no
effects on serum albumin or the ultH1zat1on of nitrates or I1p1ds 1n food.
In another study using humans, a treatment protocol was designed 1n which 12
volunteers Ingested benzole add In capsules at 1 g/day for 5 days, 1.5
g/day for the next 5 days, 2 g/day for the next 5 days and 2.5 g/day for the
last 5 days, resulting 1n a total dose of 35 g over a 20-day period (WHey
and Blgelow, 1908). Because marked symptoms of discomfort, Irritation,
weakness and malaise occurred In 9/12 volunteers the course of treatment was
not completed.
Several short-term studies 1n laboratory animals were also performed
with sodium benzoate. In a range-finding study, Smyth and Carpenter (1948)
gave groups of five male and five female Sherman rats sodium benzoate In the
diet for 30 days at dosages ranging from 16-1009 mg/kg bw/day. The
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parameters evaluated Included survival, appetite, growth rate and a limited
h1sto1og1cal examination. No adverse effects were reported at 1009
mg/kg/day, the highest dosage tested.
Griffith (1929) fed diets containing sodium benzoate at 0, 1.5, 2.0, 2.5
or 3% to young male white rats for 40 days to study Us effects on growth
rate. Food consumption was equivalent at all dietary levels. There were no
effects on growth at <2.5%. At 3%, growth was "distinctly less" than
control rats and one-third of the rats 1n this group died. The Incorpora-
tion of glyclne or gelatin In the 3/4 sodium benzoate diet resulted In normal
growth rate, but the effect on survival was not reported. In another study
of the effects of sodium benzoate 1n the diet on the growth of rats, White
(1941) observed marked stunting at 5% within 3-6 weeks. HHh the exception
of a few rats that did not tolerate the sodium benzoate, no gross signs of
toxldty were observed.
Harsh'barger (1942) pair-fed diets containing 3% sodium benzoate to
4-week-old white rats for 4-5 weeks. Marked reduction In growth rate and
food conversion efficiency were observed. In the treated groups, 2/8 rats
died. Sodium benzoate at 1% 1n the diet had no effect on growth or
survival. In another study, sodium benzoate was Incorporated Into the diets
of groups of six male and six female 23-day-old Sherman rats at 0, 2 or 5%
for 28 days (FanelH and Halllday, 1963). All high-dose rats died between
the first and second weeks, following severe CNS signs. A slight but
significant depression 1n the rate of body weight gain In males was observed
at 2%. From data provided by the Investigators, estimated equivalent doses
of sodium benzoate were 2002 and 2171 mg/kg/day for males and females,
respectively, at 2% 1n the diet.
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In a toxIcHy test with sodium benzoate, groups of five male and five
female young Sherman rats were fed diets containing sodium benzoate at 0,
1.0, 2.0, 4.0 or 8.0% for 90 days (Deuel et al., 1954). Parameters
evaluated Included food consumption, weight gain, relative liver and kidney
weight, gross pathology and microscopic pathology, apparently limited to the
liver and kidney. Experimental results from both sexes were combined. The
Investigators estimated Intakes of 0, 640, 1320, 2620 and 6290 mg/kg/day for
controls and low to high treated groups, respectively. One control rat and
one low-dose group rat and two each from the two higher dose groups died
from Infections. In addition, four treatment-related deaths occurred at 8%
sodium benzoate. Decreased rate of body weight gain and Increased relative
liver and kidney weights were also observed at 8%. "Frequent pathological
lesions" not otherwise specified were noted 1n rats on the 8% diet. One rat
with slight cloudy swelling of the liver was observed 1n each of the lower
dose groups but not 1n controls; however, the Investigators noted that this
1s a common lesion 1n rats and was not treatment-related.
6.1.2.2. CHRONIC -- In a 17-month study, groups of 25 male and 25
female young cross-bred white mice (10-15 g) were given benzole acid at 40
mg/kg/day 1n a paste before the main feeding (Shtenberg and Ignat'ev, 1970).
A control group (not otherwise specified) was maintained. Parameters of
toxldty evaluated were food consumption, weight gain, general appearance
and behavior, survival, response to stress and organ weights; however, only
stress response results were reported over the 17-month feeding period.
When subjected to a 5-day fast during which benzole add treatment was
continued by gavage administration, mortality was 50.0% compared with 12.5%
In controls, and weight loss was 26.0% of body weight 1n treated mice
compared with 17.8% In controls. Following the fast, controls regained
weight 1n only 1.6 days, compared with 2.7 days for treated mice.
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In an 18-month study with benzole add, mice were treated at 0, 40 and
80 mg/kg/day by an unspecified route for 3, 8 or 18 months (Ignat'ev, 1965).
As reported by Informatics, Inc. (1972), evaluated parameters of toxlclty
Included general appearance, survival, reproduction, food and water utiliza-
tion, weight gain, blood tests, urine tests, hlstopathology, carclnogenlclty
and response to various stressors. Depressed weight gain was reported at 80
mg/kg/day 1n both sexes. In addition, viability was decreased, organ
weights were affected and the ability of the liver to detoxify carbon tetra-
chlorlde was reduced. Data were not presented, however, and these effects
cannot be evaluated. Informatics, Inc. (1972) noted that "this study was
reported In several other papers, none of which provided data sufficient to
justify the conclusions reached."
In an 18-month study using young (100-120 g) Wlstar rats (Shtenberg and
Ignat'ev, 1970), groups of 10 males and 10 females received 40 mg/kg/day 1n
a paste before the main feeding. A control group received basal cHet alone.
Food and water consumption, body weight gain, blood tests and response to
stress were examined, but data reported were minimal. It appears that
treated males may have had reduced food and water Intake, compared with
controls. Informatics, Inc. (1972) presented minimal data from another
report from the same laboratory, 1n which there were no effects on body
weight, survival or gross or microscopic morphology of "parenchymatous
organs" from rats fed benzole add at 80 mg/kg/day for 18 months (Ignat'ev,
1965).
In a long-term growth experiment using Wlstar rats, groups of 20 females
and 30 males (5-week-old, 50-60 g) were fed diets containing 1.5% benzole
add (Marquardt, 1960). A control group consisted of 12 females and 13
males. Decreased food Intake and suppressed growth were noted up through 18
0007d -40- 05/12/87
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months of exposure. The experiment was In progress at the time of the
report. Similar results were reported 1n another experiment In which groups
of 20 male Wlstar rats and 20 male Osborne-Mendel rats were fed diets
containing benzole acid at 1.5%. The control groups consisted of 10 male
rats per strain.
A chronic toxldty-reproductlon study was performed with benzole add at
0, 0.5 or 1.0% 1n the diets of groups of 20 male and 20 female Bayer-
Elberfeld rats (Kleckebusch and Lang, 1960). The rats were pair-fed for the
first 8 weeks after which additional feeding was allowed. Hating was per-
mitted when rats were 11-12 weeks old. Benzole add feeding was continued
for 4 generations. Evaluated parameters of toxldty Included estimation of
efficiency of protein utilization. Utter size (number and weight) and
h1stolog1cal examination (not otherwise detailed) of the 4 generations.
There were no signs of toxldty over the entire llfespan of the F_ genera-
tion or 1n the 4 generations of offspring. A significant Increase In life-
span was noted; at 0.5%, some rats lived >1000 days and one rat on the "\%
diet level lived for 1346 days.
6.1.3. Other Relevant Information. For benzole acid, oral LD5Q values
of 2000 mg/kg have been reported for dogs, cats and guinea pigs (NIOSH,
1986; Sax, 1984), rabbits (Sax, 1984) and mice (FEMA, 1984). For rats,
LD5Q values range from 1050 mg/kg (Sax, 1984) to 2000-2530 mg/kg (NIOSH,
1986; FEMA, 1984). Sodium benzoate appears to be somewhat less toxic; oral
LD50s for rats are -4100 mg/kg (Sax, 1984; Wlndholz, 1983). Benzole acid
1s considered a mild skin Irritant and a severe eye Irritant 1n rabbits
(Sax, 1984).
Benzole acid and sodium benzoate have been associated with hypersensl-
t1v1ty reactions and urticaria 1n exposed persons. Clemmensen and Hjorth
0007d -41- 05/12/87
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(1982) described an outbreak of urticaria in 18/20 children following Intake
and "accidental perloral application" of mayonnaise containing benzole add.
Healthy adults reacted positively to the closed 20-m1nute patch test with
sorblc acid and benzole acid, both of which were contained In the mayon-
naise. Because the reaction 1n experimental subjects was only partially
blocked with locally applied antlhlstamlne, the authors concluded that the
reaction was due to non1mmunolog1c mechanisms and recommended no restric-
tions In the use of benzole add as a preservative 1n food.
Ros et al. (1976) and Mlchaelsson and Juhlln (1973) studied the Induc-
tion of urticaria In humans by benzene ring-containing azo-food dyes,
preservatives and drugs. Patients suffering from urticaria were given small
repeated oral doses of Individual compounds to elicit an urticaria response
(provocation test). Doses of 50, 250 or 500 mg/adm1n1strat1on were given
until a response was observed. In the Initial test with 37 patients, 22
showed a positive response to sodium benzoate (Mlchaelsson and Juhlln,
1973). Rigid dietary control resulted 1n marked Improvement 1n 61/72
urticaria patients 1n a larger more recent study (Ros et al., 1976).
Nethercott et al. (1984) observed contact urticaria 1n three workers
handling sodium benzoate In a pharmaceutical manufacturing plant. These
workers noted that physical exertion during exposure exacerbated their
reaction. The authors hypothesized that perspiration lowered the skin pH
sufficiently to convert the sodium benzoate to benzole acid. Patch tests
with sodium benzoate and benzole add In the three affected workers and In
three "control" workers revealed that benzole add was the more active
chemical.
Humans with a congenital ornlthlne carbamoyl transferase deficiency may
suffer from seizures associated with hyperammonemla resulting from an
0007d -42- 05/12/87
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Inability to convert ammonia to urea. Takeda et al. (1983) described the
successful therapeutic use of oral sodium benzoate to reduce the severity
and frequency of seizures In an 8-year-old girl. Treatment was with 200
mg/kg/day of sodium benzoate In three divided doses. Clinical and labora-
tory examination revealed no signs of toxldty. Administration of benzoate
was associated with Increased excretion of hlppurlc add, an alternative
mechanism of reducing body burden of ammonia.
Amsel and Levy (1970) orally administered sodium benzoate equivalent to
2-5 g benzole acid to healthy young male volunteers after a standardized
light breakfast. Ethanol at 50 ml In orange juice was given orally 15
minutes before or 90 minutes after the sodium benzoate. Control experiments
were carried out without ethanol. Within 1 hour of administration, ethanol
decreased urinary output of hlppurate. The Investigators determined that
the renal excretion of hlppurate was not affected by the ethanol, but that
the rate of conversion of benzo-ate to hlppurate was reduced. They suggested
that ethanol Interfered with mobilization of glydne to an available pool
and that the decrease 1n hlppurate formation occurred when readily utlHz-
able glydne was exhausted.
6.2. CARCINOGENICITY
6.2.1. Inhalation. Pertinent data regarding the Inhalation cardnogen-
1c1ty of benzole add or sodium benzoate could not be located In the avail-
able literature as cited 1n Appendix A.
6.2.2. Oral. In a cardnogenldty study, groups of 50 male and 50 female
albino Swiss mice were provided drinking water containing sodium benzoate at
2% from 39 days of age until natural death or sacrifice 1n a moribund condi-
tion (Toth, 1984). Selection of the 2% water concentration was based on the
mortality and weight loss at 4% In a 35-day test that evaluated survival,
0007d -43- 05/12/87
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body weight, chemical Intake and hlstopathology. Results of the 35-day
study at 0.5, 1, 2 and 8% were not reported. Controls In the chronic test
consisted of 100 males and 100 females. Average dally water consumption was
6.2 ml for treated males and 5.9 ma for treated females, from which the
Investigator calculated dally dosages of 124.0 mg/day for males and 119.2
mg/day for females. Assuming an average body weight of 0.030 kg, these
dosages are equivalent to 4133 and 3973 mg/kg/day for males and females,
respectively. Complete necropsies were performed on all mice, and hlsto-
pathologlc examinations of all mice Included 11 major organs and all gross
lesions. Treatment with sodium benzoate had no effect on survival or the
Incidences of any tumor types.
In another report, white cross-bred mice were given dally oral doses of
40 mg/kg benzole acid combined with 80 mg/kg sodium bisulfite 1n a paste
before the main feeding for 17 months (Shtenberg and Ignat'ev, 1970).
Malignant tumors (not otherwise specified) occurred In 8/100 mice 1n the
treated group and 1n 1/8 1n the third generations of the treated groups. No
tumors were observed 1n controls.
The NTP (1987) has not scheduled benzole add or sodium benzoate for
cardnogenlcHy testing.
6.2.3. Other Relevant Information. Olnerman and Ignat'ev (1966) Indi-
cated that exposure to 0.254 benzole acid 1n the diet Increased the suscep-
tibility of mice to 1ntraper1toneal Inoculation with Ehrllch ascltes
carcinoma cells. Inoculation of the tumor cells took place after exposure
to benzole acid for 3 months. Mice were weighed and abdominal measurements
were made before Injections and every 4 days thereafter until death or
sacrifice at 53 days for treated mice or 66 days for controls. Ascltes
tumors developed 1n 62/90 (68.8%) of benzole acid-treated mice, but only 1n
0007d -44- 05/12/87
-------
16/49 (32.6%) of basal diet-fed controls. The Investigators also reported
that carcinoma development was more Intensive and survival time was shorter
In mice treated with benzole add.
6.3. MUTAGENICITY
Benzole add and sodium benzoate have been tested for mutagenldty or
genotoxldty In prokaryotes (HcCann et al., 1975), eukaryotes (Utton
B1onet1cs, Inc., 1974) and several mammalian test systems (Litton B1onet1cs,
Inc., 1974, 1975; Olkawa et al., 1980). Results have been consistently and
unequivocally negative (Table 6-1).
6.4. TERATOGENICITY
FORL (1972) performed an oral developmental toxldty study with sodium
benzoate using CD-I mice, Wlstar rats, golden hamsters and Dutch-belted
rabbits. The compound was dissolved In water and administered by gavage to
groups of 25-30 mated mice and 24 mated rats at 0, 1.75, 8.0, 38.0 or 175
mg/kg/day on gestation days 6-15. H.lce were sacrificed for examination on
gestation day 17 and rats on gestation day 20. Hamsters (22/group) were
treated on gestation days 6-10 with dosages of 0, 3.0, 14.0, 65.0 or 300.0
mg/kg/day and sacrificed for examination on gestation day 14. In mice, rats
and hamsters, the day 1n which sperm appeared 1n a vaginal smear was desig-
nated gestation day 0. Rabbits (14-32/group) were treated at 0, 2.5, 12.0,
54.0 or 250.0 mg/kg/day on gestation days 6-18 and were killed on gestation
day 29. The day of Insemination was designated gestation day 0. For mice,
rats and hamsters, one-third of the fetuses from each Utter were examined
for visceral malformations and two-thirds were examined for skeletal malfor-
mations. Rabbit fetuses delivered live were placed In an Incubator for 24
hours to evaluate neonatal survival, after which all fetuses were evaluated
for both visceral and skeletal malformations.
0007d -45- 07/08/87
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Treatment with four dosage "levels of sodium benzoate was not associated
with maternal or fetal tox1c1ty In any of the test species. The Incidence
of visceral or skeletal anomalies was not significantly elevated In any
treated groups, nor was there any apparent difference In neonatal mortality
In rabbits.
Minor and Becker (1971) administered sodium benzoate by Intraperltoneal
Injection to groups of mated Sprague-Dawley rats (evidence of copulation
designated gestation day 1) on gestation days 9, 10 and 11 and to other
groups on gestlon days 12, 13 and 14. Dosages used 1n both series were 100,
315 and 1000 mg/kg. Controls consisted of groups receiving sodium chloride
at 90 ("low controls") or 600 mg/kg ("high controls"). In both the 9- to
11- and 12- to 14-day series, adverse effects were observed only at 1000
mg/kg. In the 9- to 11-day series, these Included gross anomalies (not
otherwise specified), reduced fetal body weight and Increased fetal death.
In the 12- to 14-day series adverse effects Included reduced fetal body
weight and Increased fetal death, but no gross anomalies. Apparently, fetal
sectioning and skeletal clearing and staining were not performed.
6.5. OTHER REPRODUCTIVE EFFECTS
In a chronic toxldty-reproductlon study (Kleckebusch and Lang, 1960)
(see Section 6.1.2.2.), groups of 20 male and 20 female Bayer-Elberfeld rats
were fed diets containing 0, 0.5 or 1.0% benzole add In a multlgeneratlon
study. There were no adverse effects on reproduction. In another study
using white rats (Peretlanu et al., 1956), benzole acid 1n the diet at
0.1-0.5 gX (<1 g/kg bw/day) had no effects on reproduction. Benzole add at
10% (>1 g/kg bw/day) produced "alterations" 1n reproduction.
0007d -47- 07/08/87
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6.6. SUMMARY
Inhalation of benzole acid was Irritating to the lungs of rats and
resulted in hlstologlcally detectable signs of Inflammation (IRDC, 1981).
Inflammatory changes were noted in all treated groups of rats exposed to 25,
250 or 1200 mg/m3, 6 hours/day, 5 days/week for 4 weeks.
Several Investigators studied the subchronlc toxldty of orally adminis-
tered benzole acid and sodium benzoate In laboratory animals and humans. A
subchronic study reported a reduced rate of body weight gain In mice with
benzole acid at 80 mg/kg/day (Shtenberg and Ignat'ev, 1970). Cats, on the
other hand, tolerated a 130-160 mg/kg/day dosage of benzole add for 23 days
without clinical signs or cUnlcopathologic evidence of liver or kidney
impairment {Bedford and Clarke, 1972).
A number of subchronlc dietary studies were performed with rats using
benzole add (Krels et al., 1967) and sodium benzoate (Smyth and Carpenter,
1948; Griffith, 1929; White, 1941; Harshbarger, 1942; Fanelll and Halllday,
1963; Oeuel et al., 1954). In the study with benzole acid, reduced growth
rate and Impaired efficiency of feed conversion were observed at 1.1X of the
diet, the only concentration tested. With sodium benzoate, mortality
occurred at dietary levels >3.0% (Griffith, 1929; Harshbarger, 1942; Fanelll
and Halllday, 1963; Oeuel et al., 1954). Depression of body weight gain was
reported for a dietary level of sodium benzoate of 2% but no mortality
{Fanelll and Halllday, 1963).
In 20- to 92-day oral studies using humans, no externally visible
adverse effects were noted at 7 or 14 mg/kg/day (Gerlach, 1909), but irrita-
tion, discomfort, weakness and malaise were observed at 25 mg/kg/day (WHey
and Blgelow, 1908). In hypersensitive humans, oral (Clemmensen and Hjorth,
0007d -48- 07/08/87
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1982; Ros et al., 1976; Mlchaelsson and JuhUn, 1973) or occupational
exposure (Nethercott et al., 1984) to benzole acid or sodium benzoate may
lead to urticaria.
Long-term oral studies using rats and mice were performed with benzole
acid. Shtenberg and Ignat'ev (1970) reported that mice treated with 40
mg/kg/day for 17 months had reduced ability to cope with stress, manifested
as an Increased Incidence of mortality and greater weight loss, compared
with controls, during a 5-day fast after the 17-month exposure period. In
another report {Ignat'ev, 1965), 80 mg/kg/day administered to mice was
associated with reduced viability and weight gain, and altered organ weights.
In rats exposed to benzole acid for >18 months, decreased food Intake
and growth were observed at 1.5X 1n the diet (Marquardt, 1960), but not at
<1.0% 1n the diet (Kleckebusch and Lang, 1960).
Data were not located regarding the Inhalation carclnogenlclty of
benzole acid or Its soluble alkali salts. In a drinking water study using
mice, exposure to 2/4 sodium benzoate for the lifetime resulted In no In-
creased Incidence of tumors (Toth, 1984). Effects of benzole add and
sodium benzoate have been consistently negative In mutagenldty tests In
prokaryotes (HcCann et al., 1975), eukaryotes (LHton Blonetlcs, Inc., 1974)
and mammalian test systems (LHton Bionetlcs, Inc., 1974; Olkawa et al.,
1980).
Oral administration of sodium benzoate appeared to cause no maternal
toxlclty, fetal toxlclty or teratogenldty 1n mice, rats, hamsters or
rabbits (FORL, 1972). The highest dosages tested were 175.0, 175.0, 300.0
and 250.0 mg/kg/day, respectively, in these species. IntraperHoneal Injec-
tion of 1000 mg/kg sodium benzoate 1n rats, however, was associated with
fetal toxlcity and gross anomalies (Minor and Becker, 1971).
C007d -49- 07/08/87
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7. EXISTING GUIDELINES AND STANDARDS
7.1. HUMAN
Benzole add and sodium benzoate have been granted GRAS status by the
FDA when used as a chemical preservative In food at concentrations of 0.1%
(FASEB, 1973; FEMA, 1984). Benzole acid Is approved for use as an anti-
microbial at concentrations ranging from 0.1-0.00001% (listed In decreasing
order of content) in condiments, relishes, sugar substitutes, Imitation
dairy products, nonalcoholic and alcoholic beverages, frozen dairy products,
fats and oils, gelatin pudding and cheese (Subcommittee on Review of the
GRAS List, 1972). Sodium benzoate 1s also approved as an antimicrobial for
use 1n foods (listed In decreasing order of content) at levels of
0.29-0.00004% 1n sweet sauces, baked goods, condiments and relishes,
processed vegetables, seasonings and flavors, jams and Jellies, fats.and
oils, gelatin pudding, confectioners frosting, processed fruit, Imitation
dairy products, gravies, nonalcoholic and alcoholic beverages, fruit Ices,
milk products, soft candy, frozen dairy products, Instant coffee and tea,.
meat products, breakfast cereals, hard candy and cheese (Subcommittee on
Review of the GRAS List, 1972).
7.2. AQUATIC
Guidelines and standards for the protection of aquatic organisms from
the effects of benzole acid could not be located 1n the available literature
as cited In Appendix A.
0007d -50- 07/08/87
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8. RISK ASSESSMENT
8.1. CARCINOGENICITY
8.1.1. Inhalation. Pertinent data regarding the Inhalation cardnogen-
icity of benzole acid or sodium benzoate could not be located in the avail-
able literature as dted 1n Appendix A.
8.1.2. Oral. Toth (1984) exposed albino Swiss mice to drinking water
containing 2% sodium benzoate from 39 days of age throughout their lifetime.
There were no effects on survival or the incidences of any tumor types. In
another study, an Increased incidence of malignant tumors was reported at a
benzole acid dose of 40 mg/kg/day for 17 months In mice (Shtenberg and
Ignat'ev, 1970), but the study was not adequately reported for critical
analysis.
8.1.3. Other Routes. Olnerman and Ignat'ev (1966) reported that mice fed
a diet containing 0.2X benzole add for 3 months were more susceptible to
Ehrllch ascltes tumor cells Injected Intraperltoneally.
8.1.4. Weight of Evidence. The negative carcinogenlcHy results from the
drinking water study by Toth (1984) constitute Inadequate evidence to
evaluate the carcinogenic potency of benzole acid In mice. The other
reports suggesting an association of carcinogenlcHy with benzole add are
considered Inadequate for evaluation. Data were not located regarding the
carcinogenic potency of benzole acid In humans. Therefore, benzole acid Is
classified as an EPA Group 0 - not classifiable as to carcinogenic potential
In humans.
8.1.5. Quantitative Risk Estimates. The lack of adequate positive data
precludes estimation of carcinogenic potencies for benzole acid or sodium
benzoate for either Inhalation or oral exposure.
0007d -51- 07/08/87
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8.2. SYSTEMIC TOXICITY
8.2.1. Inhalation Exposure.
8.2.1.1. LESS THAN LIFETIME EXPOSURES (SUBCHRONIC) -- Only one sub-
chronic inhalation study was located. IRDC (1981) described a 4-week study
in which groups of 10 young Sprague-Oawley rats/sex were exposed to benzole
acid dust at 0, 25, 250 or 1200 mg/m3, 6 hours/day, 5 days/week. H1sto-
pathological evidence of Irritation manifested as interstitial lesions of
Inflammation were observed at all exposure levels, with both the Incidence
and Intensity Increasing In a dose-related fashion. The lowest exposure may
be considered a LOAEL, but. the short exposure time does not allow for
quantitative risk assessment based on this level In this study. Data are,
therefore, considered Inadequate for derivation of a subchronlc RfD for
Inhalation exposure to benzole add.
8.2.1.2. CHRONIC EXPOSURES - Data regarding chronic Inhalation
exposure to benzole acid or sodium benzoate could not be located 1n the
available literature as cited In Appendix A. Furthermore, no criteria or
standards for occupational exposure were located and data are Insufficient
for derivation of a chronic Inhalation RfD.
8.2.2. Oral Exposure.
8.2.2.1. LESS THAN LIFETIME EXPOSURES {SUBCHRONIC) -- Several sub-
chronic oral studies have been performed with benzole add and sodium
benzoate In laboratory animals. Shtenberg and Ignat'ev (1970) reported
reduced rates of body weight gain and reduced tolerance to carbon tetra-
chlorlde poisoning in mice receiving benzole acid at oral doses of 80
mg/kg/day for 3 months. Cats, on the other hand, showed no clinical signs
of toxiclty or cllnlcopathologlcal evidence of liver or kidney Impairment
from benzole acid doses of 130-160 mg/kg/day for 23 days (Bedford and
Clarke, 1972).
0007d -52- 07/08/87
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The effects of sodium benzoate on survival and growth of rats has been
i
studied by a number of investigators (Smyth and Carpenter, 1948; Griffith,
1929; White, 1941; Harshbarger, 1942; Panel! 1 and Halllday, 1963; Deuel et
al., 1954) in dietary experiments ranging from 28-90 days. Sodium benzoate
at >3% of the diet (1500 mg/kg/day, assuming a food factor of 0.05, and
equivalent to benzole acid at 1271 mg/kg/day) resulted In mortality and
reduced growth (Griffith, 1929; Harshbarner, 1942; Fanelll and Halllday,
1963; Deuel et al.. 1954). No adverse effects In rats were reported for
sodium benzoate at dietary levels ranging from 1% (Harshbarger, 1942) to 4%
(Deuel et al., 1954). These dietary levels are equivalent to 500-2620
mg/kg/day sodium benzoate and 424-2220 mg/kg/day benzole add. The dietary
'levels associated with no adverse effects overlap with levels associated
with mortality, which suggests an unusually steep dose-response for benzole
acid and sodium benzoate.
Gerlach (1909) reported no externally visible effects In humans Ingest-
ing benzole add at 0.5-1.0 g/day Tor 44 consecutive days or after 82 doses
in 86 days or 88 doses in 92 days at 1 g/day (14 mg/kg/day). Wiley and
Bigclow (1908), however, observed Irritation, discomfort, weakness and
malaise 1n humans exposed to <1.75 g/day over a 20-day period (25 mg/kg/day).
These data Illustrate the unusually steep slope of the dose-response curve
for benzole add 1n humans. The data base for subchronlc oral exposure to
benzole acid is judged to be inadequate for quantitative risk assessment.
The chronic oral RfDs of 312 mg/day or 4 mg/kg/day for a 70 kg human for
benzole add (Section 8.2.2.2.) Is suggested as the subchronlc oral RfD for
these compounds.
8.2.2.2. CHRONIC EXPOSURES The only chronic oral data available
Involve administration of benzole add to rats and mice (Shtenberg and
0007d -53- 07/08/87
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Ignat'ev, 1970; Ignat'ev, 1965; Marquardt, 1960; Kieckebusch and Lang,
1960). A dose oT 40 rmj/kcj/day for 17 months was associated with decreased
resistance to stress in mice and possibly with reduced food and water Intake
in rats after 18 months (Shtenberg and Ignat'ev, 1970). In another report
(Ignat'ev, 1965), 80 mg/kg/day in rats for 18 months was not associated with
adverse effects on body weight, survival or gross or microscopic pathology.
In other long-term dietary studies using rats, 1.5% in the diet (750 mg/kg/
day) For 18 months was associated with decreased food Intake and growth
(Marquardt, 1960), but 1.0% of the diet (500 mg/kg/day) for lifetime
resulted In no signs of toxlclty and no adverse reproductive effects over 4
generations (Kleckebusch and Lang, 1960).
If the 40 mg/kg/day dose In the Shtenberg and Ignat'ev (1970) study, at
which mice had decreased, resistance to stress, Is considered to be the
LOAEL, application of an uncertainty factor of 1000 would result 1n an RfD
of 0.04 mg/kg/day or 2.8 mg/day for a 70 kg person. This RfD, however, Is
at the low end of the range of estimated per capita dally exposure of humans
to benzole acid and sodium benzoate.
The Subcommittee on Review of the GRAS List (1972) and FASEB (1973)
estimated possible Intakes of benzole add and sodium benzoate of 0.9-34 and
34-328 mg/day, respectively, based on a comprehensive survey of the amounts
of benzole acid and sodium benzoate produced for addition to food as a
preservative.
Benzoic acid Is a weak organic acid with a pKa of 4.2, and sodium
benzoate is highly soluble in water. In the stomach, both benzole acid and
sodium benzoate arc expected to exist principally in the Ionized form, as
benzoate. Given that both benzole acid and sodium benzoate are absorbed
rapidly and completely from the gastrointestinal tract, It seems reasonable
0007d -54- 07/08/87
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to consider dietary exposure to benzole acid and sodium benzoate as exposure
to the same compound. I.e., benzole acid. By correcting for differences In
molecular weight, 328 rmj of sodium benzoate (high end of range of estimated
daily intake) is equivalent to 278 mg benzoic acid. Adding this value to
the upper end of the range of estimated daily intake for benzoic acid of 34
mg, a total dally per capita Intake for benzole add equivalent to 312
mg/day can be estimated. Since benzoic acid and sodium benzoate are
accorded. GRAS status (FASE8, 1973; FEMA, 1984), the dally estimated Intake
of 312 mg/day for benzoic acid may be considered a human NOEL and may serve
as the basis for a chronic oral RfD. Application of an uncertainty factor
of 10 to protect sensitive Individuals Is not necessary, since sensitive
' individuals are Included in the intake values for general population.
Therefore, the chronic oral RfO for benzoic acid is 312 mg/day or 4
mg/kg/day for a 70 kg human. This RfD Is well below the oral dose of 1750
mg/day (25 mg/kg/day) of benzole acid reported to cause Irritation, dis-
comfort, weakness and malaise In humans In the early study by Wiley and
Blgelow (1908). Administration of benzoic acid to humans In this study was
by capsule, which represents a bolus dose. It 1s possible that an equiva-
lent dose administered in the diet would not cause the symptoms that
resulted from the bolus dose. This RfO can also be used for such salts of
benzole add as sodium or potassium benzoate by multiplying the RfD for
benzoic acid by the ratio of the molecular weight of the salt to benzoic
add.
Many derivatives of benzole acid that may decompose to form benzoate are
accorded GRAS status and are Incorporated as additives Into food (FEMA,
1984). The chronic oral RfD for benzole acid Is not Intended to protect
against toxlclty associated with these derivatives.
0007d -55- 07/08/87
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9. REPORTA8LE QUANTITIES
9.1. BASED ON SYSTEMIC TOXICITY
Subchrun'ic inhalation and oral and chronic oral studies in laboratory
animals and subchron'ic oral studios in humans with benzoic acid and sodium
l^cnzuatc .v c vjmmarized in Table 9-1. In Chapter 8, a chronic RfD for
benzole add was calculated given the assumption that at physiological pH,
both benzole acid and sodium benzoate are expected to exist principally as
the benzoate anlon. In Table 9-1, the dosage of benzoic add or sodium
benzoate Is expressed In terms of benzole add in mg/kg/day, and the equiva-
lent human dose 1s expressed as mg benzoic acid.
Effects attributed to chronic oral exposure to benzoic add appear to be
limited to mortality or reduced survival (RV =10} and depressed rate of
bo.dy weight gain (RV =4). In addition, Shtenberg and Ignat'ev (1970) and
Ignat'ev (1965) attributed effects on viability, weight gain, organ weights
and tolerance to stress to low levels of 'benzoic acid In rats and mice.
Because the data were Insufficiently reported, these studies were not used
In the derivation of an RQ. Human oral exposure to benzoic acid resulted In
Irritation, discomfort, weakness and malaise. Inhalation exposure to
benzoic add results 1n hlstopathologlcal lesions of Inflammation (RVe=6).
CSs for these effects, calculated for benzole acid using the data points
In Table 9-1 associated with the lowest equivalent human dose of benzoic
add for each effect, are presented In Table 9-2. CSs range from 4 (RQ of
5000) to 28 (RQ of 100) Indicating a wide range In the toxic potency of
benzoic acid under different conditions. The toxic potency of benzoic acid
appears to be much greater by Inhalation than oral exposure. In calculating
a CS for decreased growth, data from two studies using benzoic acid were
considered. In a 35-day study by Kreis et al. (1967), reduced growth and
0007d -56- 07/08/87
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Impaired food utilization were observed In rats at 550 mg/kg/day. In an
18-month study by Marquardt (1960), the dosage to rats was 750 mg/kg/day.
Either dosage results In a chronic human MED >1000 mg/day and an RV. of 1.
A CS of 10 was calculated for mortality In a 90-day study (Oeuel et al.,
1954). The CS of 28 (RQ of 100) associated with Inflammation In the lungs
by Inhalation exposure (IRDC, 1981) Is chosen to represent the toxlclty of
benzole add (Table 9-3).
9.2. BASED ON CARCINOGENICITY
CarclnogenlcHy data, summarized In Section 6.2., consist of a negative
study using albino Swiss mice exposed to sodium benzoate In drinking water
at a concentration of 2% for their natural lifespan (Toth, 1984). Data
'regarding cardnogenlclty in humans were lacking arid benzole acid was
classified In CAG Group D - not classifiable as to human carcinogenic poten-
tial. Data, therefore, were not sufficient for derivation of an F factor
and this compound Is not placed In a Potency Group. Hazard ranking for
bonzoic acid, therefore, cannot be based on carcinogenicHy.
0007d -63- 07/08/87
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TABLE 9-3
Benzole Acid
Minimum Effective Dose (MED) and Reportable Quantity (RQ)
Route:
Dose*:
Effect:
Reference:
RVd:
RVg:
Composite Score:
RQ:
Inhalation
4.2 mg/day
Inflammatory lesions In lungs
IRDC, 1981
4.6
6
28
100
*Equ1valent human dose
0007d
-64-
07/08/87
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Stuermer, O.H., D.J. NG and C.J. Morris. 1982. Organic contaminants 1n
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Subba-Rao, R.V. and M. Alexander. 1982. Effect of sorptlon on mineraliza-
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Appl. Environ. Mlcroblal. 44: 659-668.
Subba-Rao, R.V., H.E. Rubin and M. Alexander. 1982. Kinetics and extent of
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0007d -81- 07/08/87
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Sufi Ha. J.M., A. Horowitz, D.R. Shelton and J.M. Tledje. 1982. Dehalogen-
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0007d -82- 07/08/87
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0007d -83- 07/08/87
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APPENDIX A
LITERATURE SEARCHED
This HEED Is based on data Identified by computerized literature
searches of the following:
TSCATS
CASR online (U.S. EPA Chemical Activities Status Report)
TOXLINE
TOXBACK 76
TOXBACK 65
RTECS
OHM TADS
STORET
SRC Environmental Fate Data Bases
SANSS
AQUIRE
TSCAPP
NTIS
Federal Register
These searches were conducted 1n January, 1987. In addition, hand searches
were made of Chemical Abstracts {Collective Indices 5-9), and the following
secondary sources should be reviewed:
ACGIH (American ConTcrencc of Governmental Industrial Hyglenlsts).
1986. Documentation of the Threshold Limit Values and Biological
Exposure Indices, 5th ed. Cincinnati, OH.
ACGIH (American Conference of Governmental Industrial Hyglenlsts).
1986-1987. TLVs: Threshold Limit Values for Chemical Substances In
the Work Environment adopted by ACGIH with Intended Changes for
1986-1987. Cincinnati, OH. Ill p.
Clayton, G.D. and F.E. Clayton, Ed. 1981. Patty's Industrial
Hygiene and Toxicology, 3rd rev. ed., Vol. 2A. John Wiley and
Sons, NY. 2878 p.
Clayton, G.D. and F.E. Clayton, Ed. 1981. Patty's Industrial
Hygiene and Toxicology, 3rd rev. ed., Vol. 2B. John Wiley and
Sons, NY. p. 2879-3816.
Clayton, G.D. and F.E. Clayton, Ed. 1982. Patty's Industrial
Hygiene and Toxicology, 3rd rev. ed., Vol. 2C. John Wiley and
Sons, NY. p. 3817-5112.
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Grayson, M. and D. Eckroth, Ed. 1978-1984. Klrk-Othmer Encyclo-
pedia of Chemical Technology, 3rd ed. John Wiley and Sons, NY. 23
Volumes.
Hamilton, A. and H.L. Hardy. 1974. Industrial Toxicology, 3rd ed.
Publishing Sciences Group, Inc., LUtleton, MA. 575 p.
IARC (International Agency for Research on Cancer). IARC Mono-
graphs on the Evaluation of Carcinogenic Risk of Chemicals to
Humans. WHO, IARC, Lyons, France.
Jaber, H.M., W.R. Mabey, A.T. L1eu, T.W. Chou and H.L. Johnson.
1984. Data acquisition for environmental transport and fate
screening for compounds of interest to the Office of Solid Waste.
SRI International, Menlo Park, CA. EPA 600/6 81-010. NTIS
PB84-243906.
NTP (National Toxicology Program). 1986. Toxicology Research and
Testing Program. Chemicals on Standard Protocol. Management
Status.
Ouellette, R.P. and J.A. King. 1977. Chemical Week Pesticide
Register. McGraw-Hill Book Co., NY.
Sax, I.N. 1984. Dangerous Properties of Industrial Materials, 6th
ed. Van Nostrand Relnhold Co., NY.
SRI (Stanford Research Institute). 1986. Directory of Chemical
Producers. Menlo Park, CA.
U.S. EPA. 1986. Report on Status Report In the Special Review
Program, Registration Standards Program and the Data Call 1n
Programs. Registration Standards and the Data Call In Programs.
Office of Pesticide Programs, Washington, DC.
U.S. EPA. 1985. CSB Existing Chemical Assessment Tracking System.
Name and CAS Number Ordered Indexes. Office of Toxic Substances,
Washington, DC.
USITC (U.S. International Trade Commission). 1985. Synthetic
Organic Chemicals. U.S. Production and Sales, 1984, USITC Publ.
1422, Washington, DC.
Verschueren, K. 1983. Handbook of Environmental Data on Organic
Chemicals, 2nd ed. Van Nostrand Relnhold Co., NY.
Worthing, C.R. and S.8. Walker, Ed. 1983. The Pesticide Manual.
British Crop Protection Council. 695 p.
Wlndholz, M., Ed. 1983. The Merck Index, 10th ed. Merck and Co.,
Inc., Rahway, NJ.
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In addition, approximately 30 compendia of aquatic toxldty data were
reviewed, Including the following:
Battelle's Columbus Laboratories. 1971. Water Quality Criteria
Data Book. Volume 3. Effects of Chemicals on Aquatic Life.
Selected Data from the Literature through 1968. Prepared for the
U.S. EPA under Contract No. 68-01-0007. Washington, DC.
Johnson, W.W. and M.T. Flnley. 1980. Handbook of Acute Toxlclty
of Chemicals to F1sh and Aquatic Invertebrates. Summaries of
Toxldty Tests Conducted at Columbia National Fisheries Research
Laboratory. 1965-1978. U.S. Dept. Interior, F1sh and Wildlife
Serv. Res. Publ. 137, Washington, DC.
McKee, J.E. and H.W. Wolf. 1963. Water Quality Criteria, 2nd ed.
Prepared for the Resources Agency of California, State Water
Quality Control Board. Publ. No. 3-A.
Plmental, D. 1971. Ecological Effects of Pesticides on Non-Target
Species. Prepared for the U.S. EPA, Washington, DC. PB-269605.
Schneider, B.A. 1979. Toxicology Handbook. Mammalian and Aquatic
Data. Book 1: Toxicology Data. Office of Pesticide Programs, U.S.
EPA, Washington, DC. EPA 540/9-79-003. NTIS PB 80-196876.
0007d -88- 07/08/87
U.S. Environmental Protection Agency
Region V, Library
230 South Dearborn Street
Chiron. H!«nois 60604
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