methyl (2E)-{2-[(2,5-dimethylphenoxy)methyl]phenyl}(methoxyimino)ethanoate

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study performed under GLP conditions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): X-E-Oximether
- Analytical purity: >95%
- Batch No.: 26249/176
- Test substance No.: 96/288

Method

Target gene:

his- or trp-

Species / strain

Species / strain / cell type:

other: Salmonella typhimurium TA1535, TA1537, TA98, TA100, E. coli WP2 uvrA

Metabolic activation:

with and without

Metabolic activation system:

Metabolic activation was achieved by addition of S-9 mix, which consisted of the S-9 fraction from the liver of male Sprague-Dawley rats treated with AROCLOR 1254, mixed with a series of cofactors (MgCl2, KCl, Glucose-6-phosphate, NADP, phosphate buffer).

Test concentrations with justification for top dose:

20 - 5000 µg/plate

Vehicle / solvent:

Acetone

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Acetone

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2- Aminoanthracene (2-AA), N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG), 4-Nitro-o-phenylendiamine (NOPD), 9-Aminoacridine chloride monohydrate (AAC) and N-Ethyl-N'-nitro-N-nitrosoguanidine (ENNG).

Details on test system and experimental conditions:

The test was conducted according to the Ames procedure (standard plate test and preincubation test, with and without metabolic activation).

Number of plates: 3 test plates per dose or per control.

Vehicle: Acetone.

The study comprised 3 independent experiments:

1st Experiment:
- Strains: TA 1535, TA 100, TA 1537, TA 98, E. coli WP2 uvrA.
- Doses: 0, 20, 100, 500, 2500 and 5000 µg/plate.
- Test condition: Standard plate test with and without S-9 mix.

2nd Experiment:
- Strains: TA 1535, TA 100, TA 1537, TA 98, E. coli WP2 uvrA.
- Doses: 0, 20, 100, 500, 2500 and 5000 µg/plate.
- Test condition: Standard plate test (TA 100) and preincubation test (all tester strains) with and without S-9 mix.

3rd Experiment:
- Strains: TA 100
- Doses: 0, 500, 1000, 2000, 3000 and 4000 µg/plate.
- Test condition: Preincubation test with and without S-9 mix.

Positive control substance:
- With S-9 mix: 2- Aminoanthracene for the strains TA 100, TA 98, TA 1537, TA 1535 and E. coli WP2 uvrA.
- Without S-9 mix: N-Methyl-N' -nitro-N-nitrosoguanidine (strains TA 100 and TA 1535); 4-Nitro-o-phenylendiamine (strain TA 98); 9-Aminoacridine chloride monohydrate (strain 1537) and N-Ethyl-N' -nitro-N-nitrosoguanidine (strain E. coli WP2 uvrA).

Evaluation criteria:

The assessment for the test was based on the number of histidine or tryptophan revertant colonies per plate. In general, a substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results.

Results and discussion

Test results

Species / strain:

other: Salmonella typhimurium TA1535, TA1537, TA98, TA100, E. coli WP2 uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

A weakly bacteriotoxic effect (decrease in the titer) was observed in the preincubation test at doses ≥ 2500 µg/plate.

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Standard Plate Test:

Mean number of revertants with S-9 mix

Test substance	Dose/Plate (µg)	TA 1535	TA 100	TA 1537	TA 98	E. coli WP2 uvrA
Vehicle control	-	16	156	11	43	43
X-E-Oximether	20	16	1924	12	44	50
	100	14	211	12	43	53
	500	12	215	98	34	41
	2500	13	230	8	33	45
	5000	12	216	13	29	36
Positive control	2.5	114	905	114	1182	-
	60	-	-	-	-	206

Positive Control: 2 -Aminoanthracene

Mean number of revertants without S-9 mix

Test substance	Dose/Plate (µg)	TA 1535	TA 100	TA 1537	TA 98	E. coli WP2 uvrA
Vehicle control	-	13	131	7	30	42
X-E-Oximether	20	13	130	7	32	40
	100	14	130	6	34	47
	500	13	119	8	32	42
	2500	13	136	7	31	39
	5000	12	144	8	24	35
Positive control 1	5	760	1252	-	-	-
Positive control 2	10	-	-	-	958	-
Positive control 3	100	-	-	715	-	-
Positive control 4	10	-	-	-	-	651

Positive control 1: N-Methyl-N'-nitro-N-nitrosoguanidine

Positive control 2: 9-aminoacridine chloride monohydrate

Positive control 3: 4 -Nitro-o-phenylendiamine

Positive control 4: N-Ethyl-N'-nitro-N-nitrosoguanidine

Preincubation Test:

Mean number of revertants with S-9 mix

Test substance	Dose/Plate (µg)	TA 1535	TA 100	TA 1537	TA 98	E. coli WP2 uvrA
Vehicle control	-	16	13	14	38	39
X-E-Oximether	20	19	4	10	42	39
	100	22	2	12	32	41
	500	16	11	11	28	42
	2500	11	17	6	18	43
	5000	9	13	4	17	41
Positive control	2.5	125	897	106	717	-
	60	-	-	-	-	193

Positive Control: 2 -Aminoanthracene

Mean number of revertants without S-9 mix

Test substance	Dose/Plate (µg)	TA 1535	TA 100	TA 1537	TA 98	E. coli WP2 uvrA
Vehicle control	-	15	125	7	26	26
X-E-Oximether	20	18	127	8	36	23
	100	16	137	9	33	26
	500	13	126	7	19	21
	2500	13	126	6	18	21
	5000	14	121	6	13	24
Positive control 1	5	1033	1055	-	-	-
Positive control 2	10	-	-	-	1205	-
Positive control 3	100	-	-	618	-	-
Positive control 4	10	-	-	-	-	731

Positive control 1: N-Methyl-N'-nitro-N-nitrosoguanidine

Positive control 2: 9-aminoacridine chloride monohydrate

Positive control 3: 4 -Nitro-o-phenylendiamine

Positive control 4: N-Ethyl-N'-nitro-N-nitrosoguanidine

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative