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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1997-11-19 to 1997-11-28
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Read-across from guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998
Report Date:
1998

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
May 26, 1983
Deviations:
no
Qualifier:
according to
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

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent

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.
Controlsopen allclose all
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
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
without S9-mix

Migrated to IUCLID6: 80 µg/plate for TA 1537
Positive controls:
yes
Positive control substance:
other: 4-Nitro-o-phenylenediamine, 40 µg/plate for TA 1538, TA 98
Remarks:
without S9-mix
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
Species / strain:
other: TA 1535, TA 1537, TA 1538, TA 98, 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


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

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.