Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Data on benzyl alcohol:

Benzyl alcohol was not mutagenic in several Ames tests with Salmonella typhimurium TA92, TA94, TA98, TA100, TA1535, TA1537, and TA1538 with and without metabolic activation (e.g. Mortelmans et al. 1986/NTP 1989/Zeiger 1990; Ishidate et al./MHW 1984; Kuroda 1984/Yoo 1985). Negative results were reported in the mouse L5178Y/TK+/- lymphoma assay with metabolic activation, whereas positive results were obtained without metabolic activation (NTP TR343, 1989/McGregor et al. 1988). The effect was associated with cytotoxicity (effect occurred at 4500 µg/mL with rel. total cell growth 20 %, lethal dose 5000 µg/mL). Benzyl alcohol induced significant increase in chromosomal aberrations in CHO cells in the presence, but not in the absence of S9 (NTP TR343, 1989/Anderson et al. 1990); again the positive result with S9 was obtained at a relatively high dose (4000 µg/mL). A negative result of a chromosomal aberration assay was reported by Ishidate et al. (Ishidate1984/MHW). The induction of sister chromatid exchanges in CHO cells was found to be equivocal (NTP TR343, 1989/Anderson et al. 1990). A more recent in vitro micronucleus test (Fowler 2012) was negative. In that test, which was according to OECD TG 487, no toxicity and either no induction of micronuclei was noted up to the maximum test concentration of 10 mM (1081 µg/mL) recommended in the guideline.

In vivo data on benzyl alcohol are negative in different tests: a Mouse Micronucleus assay using 50, 100, 200 mg/kg benzyl alcohol by i.p. injection was negative at all doses tested; a second study with repeated i.p. injections (4 x 100 mg/kg) confirmed this result (Hayashi et al./MHW 1988). A Replicative DNA Synthesis assay using male Fischer 344 rats given a single dose of 0, 300 or 600 mg/kg bw benzyl alcohol by gavage was negative at all doses tested (Uno et al. 1994). A Replicative DNA Synthesis assay using male B6C3F1 male mice given a single dose of 0, 400 or 800 mg/kg bw benzyl alcohol by gavage was negative at all doses tested (Miyagawa et al. 1995). A Drosophila melanogaster SRL assay with benzylalcohol 5000 ppm (feed) and 8000 ppm (injection) was negative (NTP TR343/Foureman et al. 1994).

Furthermore, the negative carcinogenicity test in two species via the route shows that there is no concern for the health risks from mutations in somatic cells.

Data on other compounds of the category:

Also the other related benzyl derivatives (benzaldehyde, benzoic acid and the benzoate salts) were negative when tested in in vitro Ames tests with or without metabolic activation. Inconclusive results were obtained with other in vitro genotoxicity assays. Again, no genotoxicity was observed in the in vivo cytogenetic, micronucleus, or other assay, which leads to the conclusion: “The weight of the evidence of the in vitro and in vivo genotoxicity data indicates that these chemicals are not mutagenic or clastogenic” (OECD 2004). Moreover, all of the substances are not carcinogenic in long-term carcinogenicity studies; therefore, no evidence for mutagenic carcinogenicity exists.

Finally, it is concluded: “This evidence of safety is supported by the fact that the intake of benzyl derivatives as natural components of traditional foods is greater than their intake as intentionally added flavouring substances” (FEMA 2005).

A comprehensive document providing discussion and justification for the conclusions drawn also on this chapter is attached to this endpoint summary. Summary of the evaluations of other expert groups/regulators are provided below:

 Toxicologic programme/commission  Reference  Conclusion on genotoxicity
 EFSA, Panel on Food Contact Materials, Enzymes,Flavourings and Processing Aids (CEF SCIENTIFIC OPINIONFlavouring Group Evaluation 54, Revision 1 (FGE.54Rev1) (2009):Consideration of benzyl derivatives evaluated by JECFA (57th meeting)structurally related to benzyl alcohols, benzaldehydes, a related acetal, benzoicacids and related esters evaluated by EFSA in FGE.20Rev1 (2009) 1Scientific Opinion of the Panel on Food Contact Materials, Enzymes,Flavourings and Processing Aids (CEF). The EFSA Journal (2009) 1025, 1-73   “Conclusion on genotoxicity A total of 12 benzyl derivatives in the group have been tested for genotoxicity. In view of the mainly negative results in the assays in vitro and the uniformly negative results in well-recognised assays in vivo, the Committee concluded that the group of benzyl derivatives is not genotoxic in vivo.”
 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  “All chemicals showed no mutagenic activity in in vitro Ames tests. Various results were obtained with other in vitro genotoxicity assays. Sodium benzoate and benzyl alcohol showed no genotoxicity in vivo. While some mixed and/or equivocal in vitro chromosomal/chromatid responses have been observed, no genotoxicity was observed in the in vivo cytogenetic, micronucleus, or other assays. The weight of the evidence of the in vitro and in vivo genotoxicity data indicates that these chemicals are not mutagenic or clastogenic. They also are not carcinogenic in long-term carcinogenicity studies.”
 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  “In several in vitro tests on genotoxicity, benzoic acid and sodium benzoate tested negative. For sodium benzoate, in contrast to benzoic acid, consistently positive results were obtained in tests on sister chromatid exchange and chromosome aberrations without metabolic activation. In vivo studies for benzoic acid were not identified. For sodium benzoate, negative results were obtained in vivo in a cytogenetic assay with rats and a host-mediated assay with single or multiple oral application. However, a dominant lethal assay with rats gave a positive result. Therefore, a possible genotoxic activity of sodium benzoate cannot be ruled out entirely at present.”
 FEMA (Expert Panel of the Flavor and Extract ManufacturersAssociation) The FEMA GRAS assessment of benzyl derivatives used as flavor ingredients (2005), Food and Chemical Toxicology 43, 1207–1240  “Based on the mainly negative results in the standardized battery {i.e.,AMS (-),UDS (-),SCE (±),ABS (±), and MLA (±)} of in vitro genotoxicity assays and the uniformly negative results in well-recognized in vivo genotoxicity assays,it is concluded that the group of benzyl derivatives is not genotoxic in vivo.”“The group of benzyl derivatives discussed here was determined to be generally recognized as safe (GRAS) under conditions of intended use as flavor ingredients by the FEMA Expert Panel in 1965.”Affirmation of GRAS status in 1978 and 1993.“This evidence of safety is supported by the fact that the intake of benzyl derivatives as natural components of traditional foods is greater than their intake as intentionally added flavouring substances.“ 
 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  “The Committee also reviewed data from genotoxicity studies. None of the four compounds was mutagenic in the Ames test, either with or without metabolic activation. The compounds all induced gene mutations in the mouse lymphoma assay at the thymidine kinase locus (benzoic acid was not tested), although the requirement for metabolic activation varied. Some weak clastogenic activity was noted in in vitro assays, but not in in vivo assays.”
 US NTP (National Toxicology Programme)  US NIH (National Institutes of Health) (1989), NTP Technical Report 343 on the Toxicology and Carcinogenesis, Studies of Benzyl alcohol; NIH Publication No. 89-2599 “Benzyl alcohol was not mutagenic when tested by the preincubation protocol in the presence or absence of exogenous metabolic activation in Salmonella typhimurium strains TA98, TA100, TA1535, or TA1537. In the mouse L5178Y/TK+/- lymphoma assay, benzyl alcohol induced an increase in trifluorothymidine (Tft)-resistant cells in the absence, but not in the presence, of S9; the effect was associated with toxicity. In cytogenetic assays with Chinese hamster ovary (CHO) cells, treatment with benzyl alcohol produced an increase in sister chromatid exchanges (SCEs) which was judged to be equivocal both with and without S9; a significant increase in chromosomal aberra tions was observed after exposure to benzyl alcohol in the presence, but not the absence, of S9.” 

In addition in an Annex VI Report entitled proposal for harmonized classification and labelling; substance name: benzoic acid, prepared by the German BAuA, dated August 2011, the following conclusion is drawn: “Summary and discussion of mutagenicity: In the absence of a genotoxic potential in in vivo studies and negative results in carcinogenicity studies, no classification and labelling regarding mutagenicity is required.”


Justification for selection of genetic toxicity endpoint
No study is selected here, since several studies including human data were evaluated based on a weight of evidence approach.

Short description of key information:
Several Ames tests with S. typhimurium TA92, TA94, TA98, TA100, TA1535, TA1537, and TA1538 with and without metabolic activation yielded negative results. Inconclusive results were obtained from in vitro tests (mouse lymphoma, chromosome aberration, sister chromatid exchange assay). All in vivo tests performed with benzyl alcohol (mouse micronucleus test, Drosophila SLRL, Replicative DNA-synthesis) were negative.
Read across leads to the conclusion that the weight of the evidence of the in vitro and in vivo genotoxicity data indicate that these chemicals are not mutagenic or clastogenic. This is supported by OECD risk assessment, 2004.

Endpoint Conclusion: No adverse effect observed (negative)

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 genetic toxicity.

Based on the available data and according to criteria of Regulation (EC) No. 1272/2008, Annex I, classification for genetic toxicity would not be justified.