2-anilinoethanol

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The test substance is considered to be not genotoxic in Salmonella typhimurium and Escherichia coli.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant, guideline study, available as unpublished report no restrictions, fully adequate for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

GLP compliance:

yes (incl. QA statement)

Remarks:

Harlan Cytotest Cell Research GmbH (Harlan CCR), In den Leppsteinswiesen 19, 64380 Rossdorf, Germany

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

E. coli WP2 uvr A

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

Phenobarbital/β-naphthoflavone induced rat liver S9-mix

Test concentrations with justification for top dose:

Pre-experiment: 0; 3; 10; 33; 100; 333; 1000; 2500; and 5000 μg/plate
Main experiment: 0; 33; 100; 333; 1000; 2500; and 5000 μg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The solvent was chosen because of its solubility properties and its relative nontoxicity to the bacteria.

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

methylmethanesulfonate

other: 4-nitro-o-phenylene-diamine, 2-aminoanthracene

Remarks:

With S9: 2-aminoanthracene (all strains), Without S9: sodium azide, (TA100, TA 1535), 4-nitro-o-phenylendiamine (TA1537, TA 98), methyl methane sulfonate (WP2 uvrA)

Details on test system and experimental conditions:

METHOD OF APPLICATION: Plate Incorporation (pre-experiment/toxicity test) and Pre-Incubation (main experiment)

DURATION
- Preincubation period: 60 min (only, pre-incubation experiment)
- Exposure duration: at least 48 hours

NUMBER OF REPLICATIONS: 3 plates per concentration

DETERMINATION OF CYTOTOXICITY
- Method: Reduction in the number of spontaneous revertants or a clearing of the bacterial background lawn.

Evaluation criteria:

- A test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA 98, TA 100, and WP2 uvrA) or thrice (strains TA 1535 and TA 1537) the colony count of the corresponding solvent control is observed.
- A dose dependent increase is considered biologically relevant if the threshold is exceeded at more than one concentration.
- An increase exceeding the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment.
- A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls such an increase is not considered biologically relevant.

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

- No precipitation of the test item occurred up to the highest investigated dose.
- The plates incubated with the test item showed normal background growth up to 5000 μg/plate with and without S9 mix in all strains used.
- No toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), occurred in the test groups with and without metabolic activation in both experiments
- No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with 2-Anilinoethanol at any concentration level, neither in the presence nor absence of metabolic activation (S9 mix).
- In the pre-experiment, with metabolic activation, the number of colonies did not quite reach the lower limit of our historical control data in strain TA 98 in the solvent control. Since this deviation is rather small, this effect is judged to be based upon statistical fluctuations and has no detrimental impact on the outcome of the study.
- Appropriate reference mutagens were used as positive controls. They showed a distinct increase in induced revertant colonies.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Three genetic toxicity studies are performed. In a reverse mutation assay, performed in accordance to OECD 471 and GLP, the test substance dissolved in DMSO was tested in a plate incorporation test (Pre-experiment) and a pre-incubation test (Main experiment) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100, and the Escherichia coli strain WP2 uvrA (BASF 2015). The assay was performed with and without liver microsomal activation (Phenobarbital/β-naphthoflavone induced rat liver S9). Each concentration, including the controls, was tested in triplicate. The test item was tested at the following concentrations: Pre-experiment: 3; 10; 33; 100; 333; 1000; 2500; and 5000 μg/plate, Main Experiment: 33; 100; 333; 1000; 2500; and 5000 μg/plate. No precipitation of the test item occurred up to the highest investigated dose. The plates incubated with the test item showed normal background growth up to 5000 μg/plate with and without S9 mix in all strains used. No toxic effects, evident as a reduction in the number of revertants occurred in the test groups with and without metabolic activation in both experiments. No substantial increase in revertant colony numbers of any of the five tester strains was observed at any dose level, neither in the presence nor absence of metabolic activation. Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies. In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used. Therefore, the test substance is considered to be non-mutagenic in this Salmonella typhimurium and Escherichia coli reverse mutation assay.

 

In another OECD 471 guideline study (BASF 1991) one strain of Salmonella typhimurium (TA98) was used to test the mutagenic potential of 2-anilinoethanol both with and without metabolic activation. Additionally, norharman was used as an S9 mix additive and tested as a third group. Three standard plate tests were performed. In the first test the strain was exposed to 20, 100, 500, 5000 µg/plate. In the second test to 0, 0.8, 4, 20, 100, 500 µg/plate, and in the third test to 0, 4, 20, 100, 500, 2500 µg/plate. No bacteriotoxic effects were observed. An increased number of his+ revertants was only observed when a combination of S9 mix and norharman was used. However, norharman is a co-mutagen for aromatic amines and the result should therefore not be used for classification. It was therefore concluded that 2-anilinoethanol was not mutagenic under the conditions of the test. However, only one TA strain was tested.

 

In another study (Mattioni 2003) where binary quantitative structure-activity relationship (QSAR) models are developed, to classify a dataset of aromatic and secondary amine compounds as genotoxic or nongenotoxic based on information calculated solely from chemical structure, the SOS Chromotest was used to determine the actual genetoxic endpoints. This was performed with and without metabolic activation in E. coli modified with a lacZ report gene. 2-anilinoethanol was considered to be non-mutagenic in this study. However, very limited information on the method of study has been reported.

Justification for classification or non-classification

Based on the available information on 2-anilinoethanol, classification for genetic toxicity is not warrented in accordance with EU Classification, Labeling and Packaging of Substances and Mixtures (CLP) Regulation No. 1272/2008.