Fatty acids, C16-18, reaction products with triethanolamine, di-Me sulfate-quaternized

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1997-11-19 to 1997-11-28

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

May 26, 1983

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

December 29, 1992

Principles of method if other than guideline:

Study performance followed the old OECD TG 471, implemented in 1983. Therefore neither E. coli WP2 nor S. typhimurium TA102 were tested as required in the current OECD TG 471 from 21st July 1997.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C16-18(even numbered), reaction products with triethanolamine, di-Me sulfate-quaternized

Cas Number:

91032-11-0

Molecular formula:

n.a. (UVCB)

IUPAC Name:

Fatty acids, C16-18(even numbered), reaction products with triethanolamine, di-Me sulfate-quaternized

Constituent 2

Reference substance name:

Fatty acids, C16-18 even numbered, reaction products with triethanolamine, di-Me sulfate-quaternized

IUPAC Name:

Fatty acids, C16-18 even numbered, reaction products with triethanolamine, di-Me sulfate-quaternized

Method

Species / strain

Species / strain / cell type:

other: S. typhimurium TA 1535, TA 1537, TA 1538, TA 98, TA 100

Additional strain / cell type characteristics:

other: Histidine auxotroph

Metabolic activation:

with and without

Metabolic activation system:

S9 fraction (purchased as finished prod. from CCR (D-64380 Roßdorf/Darmstadt, Germany; Lot-No. 030797 manufact.: 1997-07-03, Proteine: 24.7 mg/ml S9-fraction). Fraction of male Wistar rat liver induced with Phenobarbital and ß-Naphthoflavone.

Test concentrations with justification for top dose:

First test: 8, 40, 200, 1000, 5000 µg/plate
Second test: 5, 10, 50, 100, 200 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The solution medium was chosen according to the solubility properties tested preliminary before start of the study.

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

without S9-mix Migrated to IUCLID6: 2 µg/plate for TA 1535, TA 100

Controls 2

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

without S9-mix Migrated to IUCLID6: 80 µg/plate for TA 1537

Controls 3

Positive controls:

yes

Positive control substance:

other: 4-Nitro-o-phenylenediamine, 40 µg/plate for TA 1538, TA 98

Remarks:

without S9-mix

Controls 4

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene, 2.5 µg/plate for TA 1535, TAS 1537, 5.0 µg/plate for TA 100, TA 1538, TA 98

Remarks:

with S9-mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 2

NUMBER OF PLATES EVALUATED: three per dose

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 groth 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 3.0).
- 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 positiv).

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>= 200 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: not noted

COMPARISON WITH HISTORICAL CONTROL DATA: yes

Applicant's summary and conclusion

Conclusions:

There was no evidence of induced mutant colonies over background in any of the tester strains in the presence or absence of mammalian metabolic activation in this study.

Executive summary:

In a reverse gene mutation assay in bacteria according to OECD guideline 471, 1983, strains TA1535, TA 1537, TA 1538, TA 98 and TA 100 of S. typhimurium were exposed to the fully saturated TEA-Esterquat. Two independent experiments were performed at concentrations of 8, 40, 200, 1000 and 5 000 µg/plate and the second experiment at concentrations of 5, 10, 50, 100, 200 µg/plate in the presence and absence of mammalian metabolic activation.

No evidence of biologically significant mutagenic activity of the test item was found in the presence and absence of metabolic activation, up to and including the limit concentration of 5000 µg/plate. Biologically significant bacteriotoxic effects were observed at concentrations of ≥ 200 µg/plate. 

The positive controls induced the appropriate responses in the corresponding strains and activity of metabolizing system was confirmed.

There was no evidence of induced mutant colonies over background.