Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Based on the available weight of evidence from studies for substances representative of the main constituents, the test sybstance, 'mono- and di- C16 PSE, K+ and C16-18-OH' is considered to be non-mutagenic in an Ames test, with and without metabolic activation.

 

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

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

In absence of genotoxicity study with the test substance, the endpoint has been assessed based on studies for substances representative of the main constituents, which can be categorised as phosphate esters (PSE; i.e., mono- and di- C16 PSE, K+) and alcohol (i.e., alcohols, C16-18). The results are presented below:

 

Constituent: PSE - read across study

A study was conducted to determine the genotoxic potential of read across substance, 'mono- and di- C16 PSE, K+ and H3PO4' (purity: 100%),using Ames test, according to OECD Guideline 471 and EU Method B13/14, in compliance with GLP. Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 and Escherichia coli strain WP2uvrA were treated with suspensions of the test substance using both the Ames plate incorporation and pre-incubation methods at up to eight dose levels (i.e., 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate), in triplicate, both with and without metabolic activation (10% liver S9 in standard co-factors). The dose range for Experiment 1 was predetermined and ranged from 1.5 to 5000 μg/plate. The experiment was repeated on a separate day (pre-incubation method) using fresh cultures of the bacterial strains and fresh test substance formulations. The dose range was amended following the results of Experiment 1 and was 15 to 5000 μg/plate. Six test substance concentrations per bacterial strain were selected in Experiment 2 in order to achieve both four non-toxic dose levels and the potential toxic limit of the test substance following the change in test methodology. The vehicle (sterile distilled water) control plates gave counts of revertant colonies generally within the normal range. All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with or without metabolic activation. Thus, the sensitivity of the assay and the efficacy of the S9-mix were validated. There was no visible reduction in the growth of the bacterial background lawn at any dose level, either in the presence or absence of metabolic activation (S9-mix), in the first mutation test (plate incorporation method) and second mutation test (pre-incubation method). A test substance precipitate (particulate in appearance) was noted at and above 1500 μg/plate, this observation did not prevent the scoring of revertant colonies. There were no increases in the frequency of revertant colonies recorded for any of the bacterial strains, with any dose of the test substance, either with or without metabolic activation (S9-mix), in Experiment 1 (plate incorporation method) and 2 (pre-incubation method). Under the study conditions, the read across substance was determined to be non-genotoxic in the Ames test, with and without metabolic activation (Envigo, 2017).

 

Constituent: Alcohol - read across study

A study was conducted to determine the genotoxic potential of read across substance, hexadecan-1-ol (purity: >95%), according to the OECD Guideline 471, using Ames test, in compliance with GLP. Two plate incorporation assays were performed, each in triplicate. Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98, and TA 100 were incubated (at 37ºC) with the test substance at concentrations 50, 150, 500, 1500 and 5000 µg/plate for 48 h, with and without metabolic activation. Aroclor 1254 induced rat liver S9 was used as the metabolic activation system. No increase in reverse mutation rate in any strain at dose levels up to 5000 µg/plate were observed. Respective positive and negative controls gave appropriate responses. Precipitation was noted at 5000 µg/plate, however, the study author concluded that this did not interfere with counting revertant colonies and the results. There was no evidence of cytotoxicity up to 5000 µg/plate with or without metabolic activation. There were no statistically significant differences between test and control plates observed. The study has met the validity criteria. Under the study conditions, the read across substance was determined to be non-mutagenic in the Ames test, with and without metabolic activation (OECD SIDS, 2006).

Based on the available weight of evidence from studies for substances representative of the main constituents, the test sybstance, 'mono- and di- C16 PSE, K+ and C16-18-OH' is considered to be non-mutagenic in an Ames test, with and without metabolic activation.

 

Justification for classification or non-classification

Overall, based on the available weight of evidence from studies for substances representative of the main constituents, the test sybstance, 'mono- and di- C16 PSE, K+ and C16-18-OH' does not warrant classification for genotoxicity according to EU CLP criteria (Regulation 1272/2008/EC).