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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

No genetic toxicity study with oleic acid, copper salt is available, thus the genetic toxicity will be addressed with existing data on the individual assessment entities copper and oleic acid. Oleic acid, copper salt is not expected to be mutagenic in bacteria, since the two moieties copper and oleic acid are not considered to have a gene mutation potential in bacteria.

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

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

No genetic toxicity study with oleic acid, copper salt is available, thus the genetic toxicity will be addressed with existing data on the individual moieties copper and oleic acid.

 

Copper

In vitro gene mutation

In a GLP study Ballantyne (1994) evaluated the mutagenic potential of Cu2+as copper sulphate pentahydrate in bacterial reverse mutation assay according to OECD guideline no. 471. The substance was tested at five different concentrations up to 800-1000 µg/plate at cytotoxic levels. Five different Salmonella typhimurium strains (TA98, TA100, TA1535, TA1537, and TA102) were tested both in presence and in absence of a metabolic activation system. No evidence for mutagenic activity in the strains tested was found.

 

Under normal physiological conditions, the concentration of free copper is extremely low in vivo and the majority of the copper is bound to ceruplasmin and albumin (See Section 5.1). In addition, cells contain high concentrations of potent antioxidants (e.g. glutathione). Therefore, the biological relevance of any in vitro observations would be uncertain where high concentration of the free ion would be available in cell culture growth medium. From reviews of public domain data (WHO, 1998; VRAR, 2008), there is conflicting evidence regarding the activity of copper in cell based assays for genotoxicity, however, due to the relevance of such studies in determining the genotoxicity potential of copper it is considered not appropriate or applicable to use these studies for copper and copper compounds.

Therefore it was considered more appropriate to review the genotoxic potential of copper and copper compounds using in vivo studies.

From the results above, copper sulphate pentahydrate, copper and other copper compounds are not considered genotoxic.

 

Oleic acid

In vitro gene mutation

A registration dossier shall contain information on the human health hazard assessment (regulation 1907/2006, Art.10). However, it is considered that the information requirements for oleic acid as laid down in annex VII to IX can be fulfilled by adaptation of the standard testing regime according to Annex XI, points 1.2. and 1.3. as presented in the following:

According to Regulation (EC) No 1907/2006 Annex V substances obtained from natural sources and not modified such as vegetable fats and oils as well as fatty acids from C6 to C24 and their potassium, sodium, calcium and magnesium salts are excluded from the obligation to register.

The substance subjected to registration is a monounsaturated C18- fatty acid. Based on this, the following endpoint is covered by publicly available data on fatty acids with the same or similar structure. Due to the structural similarity and the fact that only one hydrogenation of the double bond of oleic acid yields the saturated derivative stearic acid, most of the data are focused on stearic acid.

Based on in vitro data the HERA document on fatty acid salts (2002) reported that “fatty acids are negative in in vitro bacterial systems used in the Ames test (BIBRA, 1988; BIBRA, 1996). In addition, saturated fatty acids up to and including C12, and the unsaturated acid C18:1, have shown inhibition of the mutagenic activity of N-nitrosodialkylamines on Escherichia coli (Negishi et al. 1984). Also, fatty acids from C12 up to C19 have shown anticlastogenic effects in the chromosome aberration test (Renner, 1986)” (HERA, 2002).

“Stearic acid (C18) was tested for mutagenicity using the Ames test with Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538. Spot tests were performed using 50 mg/ml stearic acid suspensions in distilled water (50 µg/plate) with and without microsomal activation from hepatic S9 fractions from rats induced with Aroclor 1254 (50 µl/plate). Stearic acid had no mutagenic activity over background in the strains tested with and without metabolic activation (CIR, 1987)” (HERA, 2002).

“A solution of 99.9% pure oleic acid (C18) was tested in the Ames test using Salmonella typhimurium strains TA98, TA100 and TA1535. It was tested at concentrations of 1, 5, 10, 50, 100, 500, 1000 and 5000 µg/plate with and without metabolic activation and produced negative results (IUCLID, 2000e). In the Escherichia coli reverse mutation assay using E. coli strain WP2uvrA, concentrations of 1, 5, 10, 50, 100, 500 1,000 and 5,000 µg/plate, with and without activation, a solution of 99.9% pure oleic acid also produced negative results. It has also produced negative results in Saccharomyces cerevisiae and in DNA and damage repair assays using Bacillus subtilis (BIBRA, 1986; IUCLID, 2000e). Fatty acids, C18-22 produced negative results with and without metabolic activation in the Ames test at concentrations ranging between 4-1250 µg/plate using Salmonella typhimurium (IUCLID, 2000g)” (HERA, 2002).

The EFSA NDA Panel reported in their re-evaluation of fatty acids (E570) as a food additive, that “in the study by Shimuzu et al. (1985), stearic acid (60% purity) was assessed for its mutagenicity in a bacterial reverse mutation assay using S. Typhimurium strains TA98, TA100, TA1535, TA1537, TA1538 and E. coli WP2uvrA. The pre-incubation test was performed in the presence and absence of metabolic activation (liver S9 from polychlorinated biphenyl induced male rats) at concentrations of 1–1,000 µg/plate (seven concentrations) including negative, vehicle and positive control. The test substance did not induce an increase in revertant colonies compared to the concurrent negative control” (EFSA ANS Panel, 2017).

Overall, it was considered by the EFSA Panel as well as HERA that the available data did not raise a concern for genotoxicity of fatty acids and their salts.

For reference list please refer to endpoint summary of the moieties.

Oleic acid, copper salts

Oleic acid, copper salt is not expected to be mutagenic in bacteria, since the assessment entities copper and oleic acid have not shown gene mutation potential in bacteria. Further testing is not required. For further information on the toxicity of the individual moieties addressed, please refer to the relevant section in the IUCLID.

Justification for classification or non-classification

Oleic acid, copper salt is not expected to be mutagenic in bacteria, since the two assessment entities copper and oleic acid are not considered to have a gene mutation potential in bacteria. Due to the lack of data, specifically for in vitro clastogenicity in mammalian cells, no decision can be made with regard to classification as germ cell mutagen.

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