1H-Indole-6-carboxylic acid, 2,3-dihydro-3-(methoxyphenylmethylene)-2-oxo-, methyl ester, (3E)-

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:

30 August - 29 September 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD 471) and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

CD 352 XX

IUPAC Name:

CD 352 XX

Test material form:

other: solid

Details on test material:

- Name of test material (as cited in study report): CD 352 XX
- Physical state: yellow solid substance
- Analytical purity: 99.0 %
- Certificate of analysis: 24 august 2011
- Lot/batch No.: 0002
- Expiration date of the lot/batch: july 2012
- Storage condition of test material: room temperature in the dark (ambient humidity)

Method

Target gene:

S. typhimurium
TA 1537 hisC3076 rfa uvrB - Frameshift
TA 98 hisD3052 rfa uvrB pKM101 Frameshift
TA 100 hisG46 rfa uvrB pKM101 Base substitution
TA 1535 hisG46 rfa uvrB - Base substitution
TA 102 hisG428 rfa + pKM101, pAQ1 Base substitution

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

Plate test: 10, 30, 100, 300, 1000 µg/plate of test substance
Preincubation method: 10, 30, 100, 300, 600 µg/plate of test substance

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

mitomycin C

other: 2-aminoanthracene, 2-Aminofluoren

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation) and the preincubation method

DURATION
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 3 replications per concentration level

Evaluation criteria:

The assessment and interpretation of the results follows the OECD Guideline No. 471. In
addition, the historical negative control data ranges experienced in our laboratory were also
considered though for orientating purposes only.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

300 µg/plate with and without metabolic activation

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

100 µg/plate with and without metabolic activation

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS

- Precipitation: 300 µg/plate plate test and Preincubation method

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'. Remarks: Plate test

Any other information on results incl. tables

CD 352 XX did not induce an increase in the number of revertant colonies in different tester strains of S. typhimurium compared to the negative control when tested up to insoluble and/or bacteriotoxic concentrations in the plate test. Furthermore, metabolic activation by S9 mix did not alter the mutation frequency of these bacterial strains. The results were verified in an independent test using the preincubation method.

Bacteriotoxicity prohibited analysis of the number of revertants in some strains at 1000 μg/plate in the plate test.There was no indication for an increase in the number of revertants between 100 and 300 μg/plate where precipiation and signs of bacteriotoxicity were observed. Therefore, it was concluded that the concentration interval was sufficiently close and no further data are needed to confirm the Ames negative response.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with metabolic activation plate test and Preincubation method
negative without metabolic activation plate test and Preincubation method

CD 352 XX caused neither base-pair substitutions nor frameshift mutations in different
S. typhimurium strains in the absence and presence of metabolic activation system when
tested up to insoluble and bacteriotoxic concentrations. Based on these results it was
concluded, that CD 352 XX is "Ames negative".