Reaction mass of 2-hydroxyethyl oleate, oleic acid, and 2-hydroxyethyl palmitate

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

no S. typhimurium TA102 or E. coli strain was included

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C16-18, esters with ethylene glycol

EC Number:

292-932-1

EC Name:

Fatty acids, C16-18, esters with ethylene glycol

Cas Number:

91031-31-1

Molecular formula:

C18H36O3, C20H40O3, C34H66O2, C36H70O4, C38H74O2

Test material form:

not specified

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA98, TA100, TA1535, TA1537 and TA1538

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254.

Test concentrations with justification for top dose:

First and second experiment: 8, 40, 200, 1000 and 5000 µg/plate with and without metabolic activation

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Tween 80; 1:1 (w/w); dilution to desired test concentrations with water

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

untreated suspension

Negative solvent / vehicle controls:

yes

Remarks:

Tween 80/bidist. water

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: without S9: sodium azide (2 µg/plate) for TA100 and TA1535; 9-aminoacridine (80 µg/plate) for TA1537; 4-nitro-o-phenylendiamine (40 µg/plate) for TA98 and TA1538

Controls 2

Untreated negative controls:

yes

Remarks:

untreatred suspension

Negative solvent / vehicle controls:

yes

Remarks:

Tween 80/bidist. water

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: with S9: 2-aminoanthracene (2.5 µg/plate) for TA1535 and TA1537; (5 µg/plate) for TA98, TA100 and TA1538

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 replications each in 2 independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

Evaluation criteria:

A combination of the following criteria was considered as a positive result: the plate background of non-reverted bacteria did not show any growth reduction versus the respective negative controls; the spontaneous mutation rates of each tester strain per plate were within the characteristic spontaneous mutation range; as a rule, the positive controls showed mutation rates exceeding the control values of TA 100 at least by the factor 2.0 and those of the other tester strains at least by the factor 3.03; at more than one dose tested, the test substance caused at least a 2.0-fold increase in comparison with the negative controls in the tester strain TA 100. For the other tester strains, an increase in the mutation rate of more than 3.0 above the corresponding negative controls was considered positive.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: noted at concentrations of 5000 µg/plate

Any other information on results incl. tables

Table 1: Maximum revertants of both experimental series

	Maximum number of revertants (dose level in µg/mL)
	solvent control		positive control		test substance
Strain	With S9	Without S9	With S9	Without S9	With S9	Without S9
TA 98	45.0	25	1218	643	46.7 (1000)	24.3 (1000)
TA 100	132.7	111	936.7	318.0	122.3 (1000)	115.0 (1000)
TA 1535	19.0	12.3	274.0	384.0	15.0 (1000)	13.0 (200)
TA 1537	11.3	9.3	124.7	955.3	13.3 (200)	11.3 (1000)
TA 1538	18.7	9.0	1499.7	1678.0	24.3 (200)	10.7 (40)

Applicant's summary and conclusion

Conclusions:

The substance showed no evidence of mutagenic activity in this bacterial system under the test conditions employed, with and without a metabolic activation system.

Executive summary:

In the in vitro genotoxicity study (Ames test) the substance was tested for mutagenicity in Salmonella typhimurium strains TA1535, TA1537, TA 1538, TA98 and TA100. Concentrations of up to 5000 μg/plate were tested. No evidence of mutagenic activity was seen at any concentration of the substance. It was concluded that the substance showed no evidence of mutagenic activity in this bacterial system under the test conditions employed, with and without a metabolic activation system..