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Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
multi-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Maximum reliability for read across-studies Critical study in OECD risk assessment, 2004: Meets generally accepted scientific standards, well documented and acceptable for assessment
Principles of method if other than guideline:
4-generation diet-study. Cited from OECD risk assessment: "A robust protocol, according to standards at that time, was used. Taking into account the reputation of the investigators a high quality has to be assumed."
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
initial body weight:40-50 g
Route of administration:
oral: feed
Details on exposure:
oral feed (first 8 weeks paired feed technique; afterwards ad libitum)
Duration of treatment / exposure:
Exposure period: generation 1 and 2: lifelong; generation 3: 16 weeks; generation 4: until breeding
Frequency of treatment:
continuously in diet
Details on study schedule:
lifelong (4-generations)
Remarks:
Doses / Concentrations:
approx. 375 and 750 mg/kg bw (0.5 and 1 % in diet)
Basis:

No. of animals per sex per dose:
20
Control animals:
yes
Details on study design:
The mean compound consumption was calculated according to Lehman, A.J., Assoc. Food Drug Off. Q. Bull. 18, 66 (1954).
Dose descriptor:
NOAEL
Effect level:
>= 750 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: No indications for parental toxicity at highest dose of 750 mg/kg; reference: OECD risk assessment, 2004
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
>= 750 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: No indications for fetal toxicity at highest dose of 750 mg/kg; reference: OECD risk assessment, 2004
Dose descriptor:
NOAEL
Generation:
F2
Effect level:
>= 750 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: No indications for fetal toxicity at highest dose of 750 mg/kg; reference: OECD risk assessment, 2004
Reproductive effects observed:
not specified

"In all 4 generations no influence on growth (weight, weight gain and food efficiency (measured by protein efficiency) and organ weights was found. In all 4 generations, no effects on fertility ("Fortpflanzung") and lactation ("Aufzucht der Jungen") was found. The animals of the 3rd generation were sacrificed and examined histopathologically after 16 weeks (after lactation of the pups). No remarkable histopathological findings were found.

In the paper no information is given on the organs investigated, however the robustness of the total study, the reputation of the

investigators, as well as the reputation of the Professor who did the histopathologic investigation, a high quality has to be assumed.

From other parameters it can be assumed that as a minimum the brains, heart, liver, kidney, testis and were examined.

Feeding of 0.5 % led to prolongation of survival compared to controls. In addition a so called "Alters-Paarung" after 48 weeks gave no

influence on start of menopause."

Executive summary:

Cited from OECD 2004: "In a 4-generation study 20 rats/sex/group were dosed continuously by diet with 375 or 750 mg/kg/day benzoic acid. In all 4 generations, no effects on fertility (“Fortpflanzung”) and lactation (“Aufzucht der Jungen”) were found. In addition a so-called “Alters-Paarung” after 48 weeks gave no influence on start of menopauze. NOAEL (Parental) > 750 mg/kg/day, NOAEL (F1 Offspring) > 750 mg/kg/day, NOAEL (F2 Offspring) > 750 mg/kg/day."

(Kieckebusch & Lang, 1960, OECD 2004)

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Study duration:
subchronic
Species:
rat
Quality of whole database:
The endpoint conclusion is based on several regulatory assessments (e.g. OECD HPV programme), therefore a high quality has to be assumed.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

No explicit study on fertility is available for benzyl alcohol. However, histopathological examinations of reproductive tissues in repeated dose rodent-studies are of high value and high sensitivity for the evaluation of reproductive toxicity, as confirmed by literature (BAuA Forschungsbericht 984, 2003,Mangelsdorf. et al., Reg. Tox. and Pharm.37, 2003, 356-369; Ulbrich & Palmer, J. Am. Coll. Toxicol. 14, 1995, 293-327; Janer et al., Reproductive Tox. 24, 2007, 103-113; Dent, Reg. Tox. and Pharm. 48, 2007, 241-258; Sanbuissho et al., J Tox. Sci. 34, 2009, Special Issue SP1-SP22). Histopathological changes on the reproductive organs in these studies are indicative of effects on fertility, whereas the absence of such effects gives evidence that a substance does not influence fertility.

For benzyl alcohol there is no indication for treatment-related findings on reproductive organs or tissues on the basis of gross and histopathological investigations of reproductive organs performed in chronic (2 y) and subchronic (13 w) oral toxicity studies performed on rats and mice under supervision of the US National Toxicology Programme (NTP TR 343, 1989) and in a recent repeated subacute (4 w) inhalation toxicity study on rats (Roper 2010).

Moreover, there are data on reproductive toxicity/fertility available for the other members of the category. Of most value is an early four-generation study with benzoic acid (Kieckebusch & Lang, 1960; critical study in OECD risk assessment on benzoates group, 2004). Cited from OECD 2004: "In a 4-generation study 20 rats/sex/group were dosed continuously by diet with 375 or 750 mg/kg/day benzoic acid.” The OECD certifies the four-gen. study that “A robust protocol, according to standards at that time, was used. Taking into account the reputation of the investigators a high quality has to be assumed." The study revealed no effects on fertility and lactation in all 4 generations. Additionally the so-called “Alters-Paarung” after 48 weeks gave no influence on start of menopause. According to OECD 2004 the quantitative results were NOAEL (Parental) > 750 mg/kg/day, NOAEL (F1 Offspring) > 750 mg/kg/day, NOAEL (F2 Offspring) > 750 mg/kg/day.

Another study on Fertility is available for benzaldehyd. Cited from OECD 2004: "A single study was conducted to examine the potential reproductive toxicity of benzaldehyde, and the report was available as a translation from Romanian. A group of 10 rats of breeding age were given 2 mg benzaldehyde in oil (type not specified) by gavage every other day for 32 weeks, equivalent to about 5 mg/kg bw per day. Ten controls were used. Two pregnancies in each rat, one at 75 days and one at 180 days, were studied. The end-points examined included the number of pregnant females, number of offspring born, pup body weight at days 7 and 21 post partum, and pup viability.

At the end of treatment, the body weights of control and treated rats were similar: 265 g and 260 g, respectively. It was reported that fewer females in the group given benzaldehyde than in the control group became pregnant; however, no data or statistical analyses were presented. The authors concluded that treatment did not significantly modify any of the parameters studied. No further details were available. The NOAEL was about 5 mg/kg bw per day" (Sporn et al. 1967, OECD 2004).

OECD risk assessment (2004) certifies that “In addition no compound related effects on reproductive organs (gross and histopatology examination) could be found in the (sub) chronic studies in rats and mice with benzyl acetate, benzyl alcohol, benzaldehyde, sodium benzoate…” This is further evidenced by the fact that most of the subchronic dose studies available were conducted including investigations on sperm morphology and vaginal cytology (Morrissey et.al 1988)

Taking into account all available data on reproductive toxicity/fertility and on repeated dose toxicity (examinations of reproductive organs) for the benzoates group (benzyl acetate, benzyl alcohol, benzaldehyde, sodium benzoate) in a weight of evidence approach there is reasonable evidence for the lack of a reprotoxic potential.

A comprehensive document providing discussion and justification for the conclusions drawn also on this chapter is attached to this endpoint summary in the IUCLID database.

Summary of the evaluations of other expert groups/regulators are provided below:

 Toxicologic programme/Commission Reference  conclusion on fertility
 OECD HPV (High production volume) programme  OECD (Organisation for Economic Co-operation and Development) (2004)SIDS Initial Assessment Report for 13th SIAM (Bern, 7th – 9th November 2001) on Benzoates: Benzoic acid, Sodium benzoate, Potassium benzoate, Benzyl alcohol, CAS Nos. 65-85-0, 532-32-1, 582-25-2, and 100-51-6 Chapter Toxicity to Reproduction:“In conclusion: According to IPCS CICAD 26 (2000) (only evaluating benzoic acid and sodium benzoate), no clear statement on the reproductive effects can be given on basis of the Kieckebusch & Lang (1960) and Toth (1984) studies only. However, critical evaluation of the original paper of the Kieckebusch & Lang study gives confidence of an adequately performed study although it was performed many years ago. In addition, reprotoxicity studies on benzaldehyde and benzylacetate and the fact that no compound related effects on reproductive organs were found in the (sub)chronic studies with all the compounds supports the lack of reproductive potential. Therefore the available consistent data on compounds in this group (data on benzyl acetate and benzaldehyde inclusive) taken as a whole are sufficient to demonstrate the lack of reprotoxic potential.” 
 OECD HPV (High production volume) programme(IPCS) International Programme on Chemical Safety CICAD 26 (Concise International Chemical Assessment Document 26) (corrigendum 2005/ 2000) on benzoic acid and sodium benzoate, published under Joint Sponsorship of United Nations Environment Programme, International Labour Organization, and World Health Organization, Wissenschaftliche Verlagsgesellschaft mbH, D-70009 Stuttgart 10   “For benzoic acid, two limited studies gave no indication of adverse reproductive or developmental effects. With sodium benzoate, several studies on different species have been performed, and embryotoxic and fetotoxic effects as well as malformations were seen only at doses that induced severe maternal toxicity. In a dietary study in rats, a NO(A)EL of about 1310 mg/kg body weight was established. Data on its precursors support the notion that benzoic acid is unlikely to have adverse reproductive effects at dose levels not toxic to the mother.“
 SCF (Scientific Committee on Food) European Commission, Health & Consumer Protection Directorate-General (2002),Opinion of the Scientific Committee on Food on Benzyl alcohol; SCF/CS/ADD/FLAV/78 Final   “The toxicity has been studied extensively, including acute, short-term and long-term toxicity, carcinogenicity, genotoxicity and developmental toxicity.”
 JEFCA (Joint FAO/WHO Expert Committee on Food Additives) WHO (World Health Organization) (1997) Evaluation of Certain Food Additives and Contaminants, Forty-sixth report of the Joint FAO/WHO Expert Committee on FoodAdditives; WHO technical report series 868   “At its forty-first meeting (Annex 1, reference 107), the Committee recommended that a full review of benzyl acetate, benzoic acid, the benzoate salts, benzaldehyde and benzyl alcohol be performed in 1995 to determine whether reproductive toxicity/teratogenicity studies or other studies would be required.At its present meeting, the Committee reviewed data from studies on disposition and metabolism, short-term and long-term toxicity, genotoxicity, reproductive toxicity, teratogenicity and observations in humans. ““The Committee was satisfied that the data reviewed for compounds in this group were sufficient to demonstrate the lack of teratogenic, reproductive or carcinogenic potential. Consequently, the Committee concluded that further studies were not required, and the group ADI of 0-5 mg per kg of body weight as benzoic acid equivalents was maintained.”


Short description of key information:
Based on a read across assessment and taking into account all available data on reproductive toxicity/fertility and on repeated dose toxicity (examinations of reproductive organs) for the benzoates group (benzyl acetate, benzyl alcohol, benzaldehyde, sodium benzoate) in a weight of evidence approach there is reasonable evidence for the lack of a reprotoxic potential.

Justification for selection of Effect on fertility via oral route:
The highest tired study available was selected. Overall, all studies assessed in a category/read-across approach did not show evidence for reproductive toxicity/effects on fertility.

Effects on developmental toxicity

Description of key information
Based on a read across assessment the overall conclusion, supported by OECD (2004), is that the available data on developmental toxicity for the substances of benzoates group (Benzoic Acid, Sodium Benzoate, Potassium Benzoate, Benzyl alcohol) indicate no developmental toxicity.
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

For benzyl alcohol itself two studies on developmental toxicity with mice are available. Both were prepared for NIOSH (Hazelden 1983, York 1986).

In the study of Hazelden "Benzyl alcohol dissolved in distilled water was administered by gavage at a dose of 750 mg/kg bw per day to 50 CD-1 mice on days 7-14 of gestation; evidence of copulation was considered the first day of gestation. A control group of 50 animals received distilled water only. All animals were allowed to deliver their litters and nurse their pups for three days, at which time necropsies were performed. Maternal body-weight gain and mortality, mating, gestation, numbers of live and dead pups per litter, total litter weight on days 1 and 2 post partum, litter weight change between days 1 and 3 post partum, and pup survival on days 1 and 3 post partum were recorded. During the treatment period, 18 deaths were reported, all of which were attributed to treatment; a further death was reported on day 15 of gestation, the day after treatment was terminated. Clinical signs of toxicity, including hunched posture, tremors, inactivity, prostration, hypothermia, ataxia, dyspnoea, swollen or cyanotic abdomen, and piloerection, were reported in up to 20 mice during treatment. Piloerection was also reported in some animals up to day 3 post partum, but no other clinical signs were seen after the period of administration. No differences were observed in the mating or gestation indices, the total number of resorptions, the mean length of gestation, or the number of live pups per litter between treated and control groups. Maternal body weight, measured on days 4 and 7 of gestation, was not significantly different from control values; however, statistically significant reductions were reported on day 18 of gestation (P < 0.001) and on day 3 post partum (P < 0.05). Maternal body-weight gain during days 7-18 of gestation was significantly lower than that of controls (P < 0.001). Significant reductions in pup body weight were reported, including a lower mean pup weight per litter on days 1 (P < 0.01) and 3 post partum (P < 0.001), a mean litter weight change between day 1 and day 3 post partum (P < 0.05), and a mean pup weight change between days 1 and 3 post partum (P < 0.001). No differences in pup survival were observed by day 3 post partum. The authors concluded that benzyl alcohol may be a reproductive hazard, apparently on the basis of the reductions in pup body weights, an effect that was observed in conjunction with maternal toxicity evidenced by increased mortality, reduced body weights, and clinical toxicity during the period of administration. As effects were seen on the dams and fetuses at the only dose used in this study, there was no NOAEL. LOAEL = 750 mg/kg bw per day" (cited from OECD 2004).

In the study of York "Fifty female mice were given benzyl alcohol at 550 mg/kg bw per day by gavage on days 6-15 of gestation; a further 50 mice received the corn oil vehicle. All dams were allowed to deliver naturally, and pups and dams were observed until day 3 post partum, when the experiment was terminated. Body weight, clinical observations, and mortality were recorded daily throughout treatment and up to day 3 post partum. Mortality was not significantly increased in animals given benzyl alcohol over that in the control group. One treated mouse showing languid behaviour, laboured breathing, and a rough coat died, but no other deaths or clinical signs were reported. Maternal body weight and body-weight gain during treatment and up to day 3 post partum were virtually identical for treated and control animals. All other parameters examined, including gestation index, average number of live pups per litter, and postnatal survival and pup body weight on days 0 and 3 post partum, were not significantly different from the control values. The authors concluded that, at the predicted LD10, benzyl alcohol had no significant effects on the development of CD-1 mice. NOAEL = 550 mg/kg bw per day" (cited from OECD 2004).

Data on other compounds of the category:

Additionally, there are studies on developmental toxicity available with benzoic acid on rat and hamster and with sodium benzoate on rat, mouse, hamster and on the non-rodent species rabbit.

Rats:

Pregnant Wistar rats were treated on day 9 of gestation with one dose of 510 mg/kg benzoic acid. Animals were sacrificed on Day 20 of gestation and the uterus observed in situ for implantation and resorption sites. Live fetuses were removed, examined for gross malformations, weighed, and prepared for histopathological examination. NOAEL Maternal toxicity: 510 mg/kg bw; NOAEL Teratogenicity: 510 mg/kg bw (Kimmel et al., 1971)

A study using pregnant Wistar rats, dosed with 1.75, 8, 38 or 175 mg/kg sodium benzoate by gavage on Days 6-15 of gestation showed no effect on nidation or on maternal or fetal survival; the number of abnormalities of soft and skeletal tissues did not differ from controls. NOAEL Maternal toxicity: 175 mg/kg bw; NOAEL Teratogenicity: 175 mg/kg bw (FDA PB# 221777, 1972)

A study using pregnant Wistar rats, dosed with 700, 1400, 2800, 5600 mg/kg sodium benzoate in the diet during the entire gestation showed no statistical difference in organ and bone abnormalities of fetuses between experimental groups and controls; growth of treated offsprings was similar to controls in rats dosed with 1400 mg/kg/day; reduced food intake and decreased body weight of the pregnant rats especially in the 5600 mg/kg group; 100% perinatal death rate; organ abnormalities of fetuses involved eye, brain and kidneys, in addition abnormalities of the skeletal system were found in rats dosed with >2800 mg/kg/day. The authors concluded that the effects on the dams and fetuses at the 2800 and 5600 levels were due to reduced maternal feed intake in these groups, leading to malnutrition, NOAEL Maternal toxicity: 1400 mg/kg bw; NOAEL Teratogenicity: 1400 mg/kg bw (Onodera et al., 1978)

Mice:

A study using pregnant CD-1 mice, dosed with 1.75, 8, 38 or 175 mg/kg sodium benzoate by gavage on Days 6-15 of gestation showed no effect on nidation or on maternal or fetal survival; the number of abnormalities of soft and skeletal tissues did not differ from controls. NOAEL Maternal toxicity: 175 mg/kg bw; NOAEL Teratogenicity: 175 mg/kg bw; (FDA PB# 221777, 1972)

Rabbits:

A study using pregnant Dutch-belted rabbits, dosed with 2.5, 12, 54 or 250 mg/kg sodium benzoate by gavage on Days 6-18 of gestation showed no effect on nidation or on maternal or fetal survival; the number of abnormalities of soft and skeletal tissues did not differ from controls. NOAEL Maternal toxicity: 250 mg/kg bw;NOAEL Teratogenicity: 250 mg/kg bw; (FDA PB# 221777, 1972)

Hamster:

A study using pregnant Golden hamsters, dosed with 3, 14, 65 or 300 mg/kg sodium benzoate by gavage on Days 6-10 of gestation showed no effect on nidation or on maternal or fetal survival; the number of abnormalities of soft and skeletal tissues did not differ from number in controls. NOAEL Maternal toxicity: 300 mg/kg bw; NOAEL Teratogenicity: 300 mg/kg bw (FDA PB# 221777, 1972).

Overall, OECD 2004 concluded on developmental toxicity: “Many of these studies were done by gavage (leading to greater toxicity due to the “bolus effect”). In these studies NOEL of >= 500 mg/kg were found. Thus, studies on reproductive and/or developmental toxicology performed by the administration of the benzyl compounds by gavage are likely to reveal changes at lower doses compared to studies where the substances are applied in the diet, leading to a distribution in the body over time. In conclusion: The compounds exhibit no developmental toxicity and a NOEL of 500 mg/kg/day can be established for developmental effects for this group of substances” (cited from OECD 2004).

Conclusion on developmental toxicity:

For benzyl alcohol itself two studies on developmental toxicity with mice are available. There are studies on developmental toxicity available for benzyl alcohol and different benzoates on mice, rats, hamsters and rabbits. Based on individual studies and the whole database benzyl alcohol is not toxic to development in rodents and non-rodents.

A comprehensive document providing discussion and justification for the conclusions drawn also in this chapter is attached to this endpoint summary in the IUCLID database.

Summary of the evaluations of other expert groups/regulators are provided below:

 Toxicologic programme/Commission Reference  conclusion on developmental toxicity
 OECD HPV (High production volume) programme  OECD (Organisation for Economic Co-operation and Development) (2004)SIDS Initial Assessment Report for 13th SIAM (Bern, 7th – 9th November 2001) on Benzoates: Benzoic acid, Sodium benzoate, Potassium benzoate, Benzyl alcohol, CAS Nos. 65-85-0, 532-32-1, 582-25-2, and 100-51-6 Chapter Toxicity to Reproduction:“In conclusion: According to IPCS CICAD 26 (2000) (only evaluating benzoic acid and sodium benzoate), no clear statement on the reproductive effects can be given on basis of the Kieckebusch & Lang (1960) and Toth (1984) studies only. However, critical evaluation of the original paper of the Kieckebusch & Lang study gives confidence of an adequately performed study although it was performed many years ago. In addition, reprotoxicity studies on benzaldehyde and benzylacetate and the fact that no compound related effects on reproductive organs were found in the (sub)chronic studies with all the compounds supports the lack of reproductive potential. Therefore the available consistent data on compounds in this group (data on benzyl acetate and benzaldehyde inclusive) taken as a whole are sufficient to demonstrate the lack of reprotoxic potential.” 
 OECD HPV (High production volume) programme(IPCS) International Programme on Chemical Safety CICAD 26 (Concise International Chemical Assessment Document 26) (corrigendum 2005/ 2000) on benzoic acid and sodium benzoate, published under Joint Sponsorship of United Nations Environment Programme, International Labour Organization, and World Health Organization, Wissenschaftliche Verlagsgesellschaft mbH, D-70009 Stuttgart 10   “For benzoic acid, two limited studies gave no indication of adverse reproductive or developmental effects. With sodium benzoate, several studies on different species have been performed, and embryotoxic and fetotoxic effects as well as malformations were seen only at doses that induced severe maternal toxicity. In a dietary study in rats, a NO(A)EL of about 1310 mg/kg body weight was established. Data on its precursors support the notion that benzoic acid is unlikely to have adverse reproductive effects at dose levels not toxic to the mother.“
 SCF (Scientific Committee on Food) European Commission, Health & Consumer Protection Directorate-General (2002),Opinion of the Scientific Committee on Food on Benzyl alcohol; SCF/CS/ADD/FLAV/78 Final   “The toxicity has been studied extensively, including acute, short-term and long-term toxicity, carcinogenicity, genotoxicity and developmental toxicity.”
 JEFCA (Joint FAO/WHO Expert Committee on Food Additives) WHO (World Health Organization) (1997) Evaluation of Certain Food Additives and Contaminants, Forty-sixth report of the Joint FAO/WHO Expert Committee on FoodAdditives; WHO technical report series 868   “At its forty-first meeting (Annex 1, reference 107), the Committee recommended that a full review of benzyl acetate, benzoic acid, the benzoate salts, benzaldehyde and benzyl alcohol be performed in 1995 to determine whether reproductive toxicity/teratogenicity studies or other studies would be required.At its present meeting, the Committee reviewed data from studies on disposition and metabolism, short-term and long-term toxicity, genotoxicity, reproductive toxicity, teratogenicity and observations in humans. ““The Committee was satisfied that the data reviewed for compounds in this group were sufficient to demonstrate the lack of teratogenic, reproductive or carcinogenic potential. Consequently, the Committee concluded that further studies were not required, and the group ADI of 0-5 mg per kg of body weight as benzoic acid equivalents was maintained.”


Justification for selection of Effect on developmental toxicity: via oral route:
No study selected here, since studies in mice, rats, hamsers and rabbits are relevant for the assessment of developmental toxicity based on a category/read-across approach.

Justification for classification or non-classification

According to legal classification (Directive 67/548/EEC and Regulation (EC) No. 1272/2008) benzyl alcohol is not classified for reproductive toxicity.

Based on the available data and according to criteria of Regulation (EC) No. 1272/2008, Annex I, classification as reproductive toxicant with respect to fertility or developmental toxicity would not be justified.