Alcohols, C14-18 and C16-18-unsatd.

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 March 2015 - 12 March 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to OECD and EC guidelines and in compliance with GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Unjecol 85AN

IUPAC Name:

Unjecol 85AN

Test material form:

other: Liquid

Details on test material:

- Name of test material (as cited in study report): Unjecol 84AN
- Physical state: Liquid
- Analytical purity: 98%
- Lot/batch No.: 5253
- Expiration date of the lot/batch: 30 October 2015
- Storage condition of test material: Room Temperature

Method

Target gene:

Salmonella typhimurium: histidine
Escherichia coli: tryptophan

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Test concentrations with justification for top dose:

Test 1: 5, 15, 50, 150, 1500, 5000 µg/plate
Test 2: 5, 15, 50, 150, 1500, 5000 µg/plate

Vehicle / solvent:

- Vehicle used: DMSO
- Justification for choice: The solubility of UNJECOL 85AN was assessed at 50 mg/mL in water and in DMSO. It was found to be insoluble in water but
dissolved in DMSO.

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

In the absence of S9 mix for strains TA100 and TA1535

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

In the absence of S9 mix for strain TA1537

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

In the absence of S9 mix for strain TA98

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

In the absence of S9 mix for strain WP2 uvrA (pKM101)

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene

Remarks:

In the presence of S9 mix for strains TA100, TA1535 and WP2 uvrA (pKM101)

Controls 6

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

In the presence of S9 mix for strains TA98 and TA1537

Details on test system and experimental conditions:

Method of application: Bacterial culture and agar containing histidine, biotin and tryptophan, mixed and overlaid onto prepared Petri dishes containing agar.

Duration
Test 1: Incubated at approximately 37ºC for 72 hours
Test 2: 30 minute pre-incubation at 37ºC
Test 2 - repeat: As unclear results were seen in the second test with strain TA100 in the absence and presence of S9 mix and with strain TA1535 in the presence of S9 mix, the test was repeated using the same method as in the second test.

Number of replications: 3

Assay validity: After this period, the appearance of the background bacterial lawn was examined and revertant colonies counted using an automated colony counter. Some plates were scored manually because of the presence of precipitate.
Any toxic effects of the test substance may be detected by a substantial reduction in mean revertant colony counts, by a sparse or absent background bacterial lawn, or both.

Evaluation criteria:

If exposure to a test substance produces a reproducible increase in mean revertant colony numbers of at least twice (three times in the case of strains TA1535 and TA1537) that of the concurrent vehicle controls, with some evidence of a positive concentration-response relationship, it is considered to exhibit mutagenic activity in this test system. If exposure to a test substance does not produce a reproducible increase in mean revertant colony numbers, it is considered to show no evidence of mutagenic activity in this test system. No statistical analysis is performed.
If the results obtained fail to satisfy the criteria for a clear “positive” or “negative” response, even after additional testing, the test data may be subjected to analysis to determine the statistical significance of any increases in revertant colony numbers. The statistical procedures used are those described by (Mahon et al 1989) and are usually Dunnett’s test followed, if appropriate, by trend analysis. Biological importance will be considered along with statistical significance. In general, treatment-associated increases in mean revertant colony numbers below two or three times those of the vehicle controls (as described above) are not considered biologically important. It should be noted that it is acceptable to conclude an equivocal response if no clear results can be obtained.
Occasionally, these criteria may not be appropriate to the test data and, in such cases, the Study Director would use his/her scientific judgement.

Statistics:

None stated.

Results and discussion

Test results

Test results 1

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:

not valid

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not valid

Positive controls validity:

valid

Additional information on results:

First test
No evidence of toxicity was obtained following exposure to UNJECOL 85AN. Precipitate was observed on all plates containing UNJECOL 85AN at 500, 1500 and 5000 μg/plate in the absence of S9 mix and at 1500 and 5000 μg/plate in the presence of S9 mix. A maximum exposure concentration of 5000 μg/plate was, therefore, selected for use in the second test. No substantial increases in revertant colony numbers over control counts were obtained with any of the tester strains following exposure to UNJECOL 85AN at any concentration up to and including 5000 μg/plate in either the presence or absence of S9 mix.

Second test
Toxicity and inconsistent colony counts were observed in strain TA100 in the absence and presence of S9 mix and in strain TA1535 in the presence of S9 mix, therefore, an additional test was performed. No evidence of toxicity was obtained with any other strains following exposure to UNJECOL 85AN. Precipitate was observed on all plates containing UNJECOL 85AN at 500, 1500 and 5000 μg/plate in the absence of S9 mix and at 1500 and
5000 μg/plate in the presence of S9 mix.
No substantial increases in revertant colony numbers over control counts were obtained with any of the tester strains following exposure to UNJECOL 85AN at any concentration up to and including 5000 μg/plate in either the presence or absence of S9 mix.

Second test - repeat
No evidence of toxicity was obtained following exposure to UNJECOL 85AN. Precipitate was observed on all plates containing UNJECOL 85AN at 500, 1500 and 5000 μg/plate in the absence of S9 mix and at 1500 and 5000 μg/plate in the presence of S9 mix.
No substantial increases in revertant colony numbers over control counts were obtained with any of the tester strains following exposure to UNJECOL 85AN at any concentration up to and including 5000 μg/plate in either the presence or absence of S9 mix.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

It was concluded that UNJECOL 85AN showed no evidence of mutagenic activity in this bacterial system under the test conditions employed.