1H-Indene-1,3(2H)-dione, 2-(2-quinolinyl)-, sulfonated, sodium salts

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

- Bacteria reverse mutation assay (with and without metabolic activation): negative.

- In vitro mammalian cell micronucleus test (with and without metabolic activation): negative.

- In vitro gene mutation study in mammalian cells (with and without metabolic activation): negative.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Several studies are available for the evaluation of the genotoxicity potential of the substancein vitro.

Three reverse mutation studies in bacteria are available. All three studies were performed with and without metabolic activation. In a study performed after 1997 and according to the OECD Guideline 471, the genotoxic potential of the substance was evaluated in S. typhimurium TA 98, TA 100, TA 1535, TA 1537 and E. coli WP2 uvr A strains at dose concentrations ranging from 33 to 5000 µg/plates(SCCNFP, 2004). In another study reported by Blevins et al. (Blevins R. D., 1982), the substance was tested on S. typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538 strains at only one dose level equal to 50 µg/plate. In the study reported by Hollstein et al (Hollstein M., 1978), the reverse mutation study was performed on TA 98, TA 100, TA 1535, and TA 1537 strains at three dose levels (10 µg, 100 µg, and 1 mg per plate). Under the conditions of the three studies, the substance did not show mutagenic potential.

An in vitro mammalian cell micronucleus test (Research Toxicology Centre S.p.A, 2013) was performed on the substance according to the OECD 487 using Chinese hamster lung fibroblast (V79) and dose concentrations equal to 5000, 3850, 2960, 2280, 1750, 1350, 1040, 797 and 613 μg/mL. Under the conditions of the test, the substance did not induce micronuclei neither in presence nor in absence of S9 metabolic activation.

The potential of the test item to induce mutagenicity/clastogenicity in mammalian cells in vitro was evaluated with a mouse lymphoma assay following the OECD Guideline 476 using the thymidine kinase target gene (SCCNFP, 2004). The study was conducted using concentrations of the test item in the range 118-3800 µg/ml, with and without S9 metabolic activation. Under the conditions of the test, the substance did not induce mutagenicity/clastogenicity in mouse lymphoma cells.

Moreover, the substance was tested using the in vitro micronucleus assay and the comet assay in two cell types: human lymphocytes and Vicia faba root cells. The substance was tested only in the absence of metabolic activation, at two concentrations (86.7 and 867 μg/mL), both showing some cytotoxicity. A statistically significant and concentration-dependent response was reported in the micronucleus assay in both cell systems, while the comet assay was positive only at the highest concentration (Macioscek V.K. and Kononowicz A.K, 2004). The significance of these results is questionable. No information on the purity of the test item is reported. The assay on Vicia faba is not a standard test and the experiments on human lymphocytes were conducted before the publication of the relevant OECD Guideline and shows several deficiencies in the experimental design (no GLP experiment) and in the data reporting. Furthermore, several oral long-term carcinogenicity studies with the substance revealed no evidence of carcinogenicity (SCCNFP, 2004) (WHO, 2017) (EFSA, 2009). EFSA Panel 2009 (EFSA, 2009) therefore considered that the results of Macioszek and Kononowicz were of uncertain biological significance.

An in vivo mammalian erythrocyte micronucleus test was performed on NMRI mice. The substance was administered at the dose levels of 500, 1000 and 2000 mg/kg. The treatment lasted 24 hours for the 500 and 1000 mg/kg doses and 48 hours for the 2000 mg/kg dose. The substance did not induce micronuclei in a frequency higher than the deionized water-treated animals (vehicle control). Nevertheless, the SSCNFP Committee (SCCNFP, 2004) considered the study inadequate for the evaluation as there is no demonstration that the substance has reached the target cells.

 

- Blevins R. D. (1982). Mutagenicity screening of twenty‐five cosmetic ingredients with the salmonella/microsome test. Journal of Environmental Science and Health . Part A: Environmental Science and Engineering: Toxic/Hazardous Substances and Environmental Engineering, 1982, 17(2), 217-239.

- EFSA. (2009). Scientific Opinion on the re-evaluation of Quinoline Yellow (E 104) as a food additive.

- Hollstein M. (1978). Quinoline: Conversion to a Mutagen by Human and Rodent Liver. Journal of the National Cancer Institute, 1978, 60(2), 405-410.

- Macioscek V.K. and Kononowicz A.K. (2004). The evaluation of the genotoxicity of two commonly used food colours: Quinoline Yellow (E 104) and Brilliant Black BN (E 151). Cellular and Molecular Biology Letters, 9(1):107-122.

- Research Toxicology Centre S.p.A. (2013). E104 Quinoline yellow in vitro micronucleus test in chinese hamster V79 cells.

- SCCNFP. (2004). Opinion of the scientific committee on cosmetic products and non-food products intended for consumers concerning Acid Yellow 3 - Colipa n° C54.

- WHO. (2017). Safety Evaluation of Certain Food Additives Prepared by the eighty-second meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA). WHO FOOD ADDITIVES SERIES: 73, World Health Organization, Geneva, 2017.

 

Justification for classification or non-classification

The susbtance resulted to be not genotoxic in the bacteria reverse mutation assay (with and without metabolic activation), in the in vitro mammalian cell micronucleus test (with and without metabolic activation) and in the in vitro gene mutation study in mammalian cells (with and without metabolic activation. In the light of the above, the substance shall not be classified for genetic toxicity according to the CLP Regulation (EC) No. 1272/2008.