Reaction products resulting from esterification of sucrose with saturated C16-18 (even numbered) fatty acids

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

genotoxicity in bacteria: negative with the reaction product

genotoxicity in mammalian cells: not expected based on information for the constituents

chromosome aberration: not expected based on information for the constituents

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

A study was performed to investigate the potential of the reaction product to induce gene mutations according to the plate incorporation test (experiment I) and the pre-incubation test (experiment Il) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100, and TA 102.

The assay was performed in two independent experiments both with and without liver microsomal activation. Each concentration, including the controls, was tested in triplicate. The test item was tested at the following concentrations: 33; 100; 333; 1000; 2500; and 5000 µg/plate

The plates incubated with the test item showed normal background growth up to 5000 µg/plate with and without S9 mix in aIl strains used. Minor toxic effects, evident as a reduction in the number of revertants, occurred in strains TA 98 (Exp. Il, with S9 mix) at 5000 µg/plate and in TA 102 (with and without S9 mix) at 5000 µg/Plate (Exp. I) and at 2500 and 5000 µg/plate (Exp. Il). No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with the test item at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance. Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies.

Further information for Sucrose Stearate:

In  agreement  with  other  studies,  sucrose  esters  of  fatty  acids  did  not  cause  significant  toxicological  effects, including short-term (Mitsubishi, 1991) and long-term toxicity and carcinogenicity (Mitsubishi 1994c;  Takeda  and  Flood,  2002).  From  a  2-year  chronic  toxicity/carcinogenicity  study  in  rats  a  NOAEL could be established at 5% sucrose esters of fatty acids (stearic: palmitic acids = 70:30, with a 10%  content  of  tetra-  and  higher  esters)  in  the  diet,  equal  to  1970  mg/kg  bw/day  in  males  and  2440  mg/kg bw/day in females (Mitsubishi 1994c). (taken from EFSA Journal 2010; 8 (3) 1512). Genotoxicity in mammalian cells and chromosome aberrations are not expected and a generation of further data is not necessary.

Further data for Fatty acids, C16 -18, Methyl Ester:

The mutagenic activity of ethyl linoleate (CAS 544-35-4), a homolog of Fatty acid, C16 -18, Methyl ester, was evaluated in an in vitro mammalian cell gene mutation test according to OECD guideline 476 with L5178Y mouse lymphoma cells (Verspeek-Rip, 2010). Two independent experiments (with 3 or 24 hours of exposure) were performed in the absence and presence of S9-mix with test substance concentrations up to 300 μg/mL dissolved in dimethyl sulfoxide. At this dose level cytotoxicity occurred in the presence and absence of metabolic activation. No significant increase in mutation frequency occurred in any of the test conditions, indicating that ethyl linoleate is not mutagenic in the mammalian cells in vitro.   In summary, based on a weight of evidence approach, all reliable studies consistently showed no treatment-related effects on genetic toxicity.  

The ability of ethyl linoleate (CAS 544-35-4) to induce chromosome aberrations in cultured peripheral human lymphocytes was tested according to OECD guideline 473 (Verbaan, 2010) in two independent experiments. Test substance concentrations of up to 800 µg/mL dissolved in DMSO were tested in the presence and absence of metabolic activation. At concentrations of 333 µg/mL and higher precipitation of test substance occurred. The number of cells with chromosome aberrations found in the solvent control cultures was within the laboratory historical control data range. Positive control chemicals, mitomycin C and cyclophosphamide, both produced a statistically significant increase in the incidence of cells with chromosome aberrations, indicating that the test conditions were adequate and that the metabolic activation system (S9-mix) functioned properly. Ethyl linoleate did not induce a statistically significant or biologically relevant increase in the number of cells with chromosome aberrations in the absence and presence of S9-mix, in either of the two independently repeated experiments. No effects on the number of polyploid cells and cells with endoreduplicated chromosomes were observed.

Fatty Acids, C16-18 Methyl Ester and the test substances belong to the short chain methyl esters category (SCAE Me). A detailed justification for the grouping and read-across is provided in the Justification document (see Section 13).

Further data for Fatty Acids, C16 -18:

Fatty acids are negative inin vitrobacterial 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 onEschericha coli(Negishiet al.1984). Also, fatty acids from C12 up to C19 have shown anticlastogenic effects in the chromosome aberration test (Renner, 1986).

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

Furthermore, there is no association between the normal intake of large amounts of fatty acids in the diet and mutagenicity.

 

BIBRA (1988) Toxicity Profile. n-Octanoic acid and its sodium and potassium salts. The British Industrial Biological Research Association. Surrey, UK.

BIBRA (1996) Toxicity Profile. n-Decanoic acid (and its sodium and potassium salts). The British Industrial Biological Research Association. Surrey, UK.

CIR (1987) Final report of the safety assessment for oleic acid, lauric acid, palmitic acid, myristic acid, stearic acid. Prepared by the Expert Panel of the Cosmetic Ingredient Review, Washington D.C.

Negishi, T. and Hayatsu, H. (1984) Inhibitory effect of saturated fatty acids on the mutagenicity of N-nitrosodimethylamine.Mutation Research135: 87-96.

Renner, H.W. (1986) The anticlastogenic potential of fatty acid methyl esters.Mutation Research172: 265-269.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on the results the substance is not considered to be classified for genetic toxicity under Regulation (EC) No 1272/2008, as amended for the sixth time in Regulation No 1297/2014.