Glycine, N-methyl-, N-coco acyl derivs., sodium salts

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:

2006-02-15 to 2006-03-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umweltschutz und Gewerbeaufsicht

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Glycine, N-methyl-, N-coco acyl derivs., sodium salts

EC Number:

263-193-2

EC Name:

Glycine, N-methyl-, N-coco acyl derivs., sodium salts

Cas Number:

61791-59-1

Molecular formula:

Not applicable

IUPAC Name:

disodium 2-(N-methyldodecanamido)acetate 2-(N-methyltetradecanamido)acetate

Test material form:

not specified

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 97a, TA 98, TA 100, TA 102 and TA 1535

Additional strain / cell type characteristics:

other: histidin-deficient

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 experiment:
- 1.5, 5, 15, 50, 149 µg/plate with and without metabolic activation

Second experiment:
- 37.5, 75, 150 µg/plate with and without metabolic activation

Vehicle / solvent:

water

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other:

Remarks:

NPD: 80 μg/plate in water, -S9, TA 97a, TA 98, TA 102; Sodium azide: 6 μg/plate in water, -S9, TA100 and TA1535; 2-AA: 3 μg/plate in DMSO, +S9, TA 98; BaP: 40 μg/plate in DMSO, +S9, TA 97a, TA 100, TA 102, TA 1535

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION
- Preincubation period: 12 h
- Exposure duration: 48 h

NUMBER OF REPLICATIONS:
- dublicates each in two independent experiments.

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

Evaluation criteria:

The test material may be considered positive in this test system if the following
criteria are met:
For the test item to be considered mutagenic, two-fold (or more) increases in mean revertant numbers must be observed at one strain with or without S9 mixture or there must be evidence of a dose-response relationship showing increasing numbers of mutant colonies with increasing dose levels

Statistics:

Mean values and standard deviation were calculated

Results and discussion

Test results

Key result

Species / strain:

other: S. typhimurium: TA 97a, TA 98, TA 100, TA 102 and TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

above 150 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:
- The highest concentration used in the study was 150 µg/plate, based on the cytotoxicity at higher concentrations.

Any other information on results incl. tables

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 2 plates ± Standard deviation)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	TA102	TA98	TA97a
–	0	114 ± 49	9 ± 2.4	9 ± 2.4	12 ± 0.5	130 ± 34.5
–	1.5	71 ± 15	6 ± 4	164 ± 25	8 ± 1	124 ± 36
–	5	97 ± 22	7 ± 2	125 ± 9	8 ± 1	97 ± 38
–	15	86 ± 12	6 ± 2	106 ± 19	6 ± 3	80 ± 7
–	50	90 ± 16	8 ± 1	101 ± 10	4 ± 2	83 ± 31
–	150*	113 ± 41	8 ± 3	139 ± 7	14 ± 6	120 ± 55
Positive controls, –S9	Name	NaN3	NaN3	NPD	NPD	NPD
	Concentrations (μg/plate)	6	6	80	80	80
	Mean No. of colonies/plate (average of 2 ± SD)	366 ± 101	1001 ± 0	657 ± 122	205 ± 98	1001 ± 0
+	0	138 ± 38	6 ± 2.6	158 ± 27.9	15 ± 2.4	101 ± 26.9
+	1.5	74 ± 13	5 ± 2	154 ± 44	12 ± 7	93 ± 42
+	5	127 ± 22	7 ± 1	156 ± 26	9 ± 6	115 ± 38
+	15	104 ± 14	8 ± 3	146 ± 9	8 ± 5	86 ± 31
+	50	103 ± 17	11 ± 3	159 ± 23	12 ± 7	101 ± 21
+	150*	78 ± 9	9 ± 1	115 ± 32	8 ± 5	102 ± 31
Positive controls, +S9	Name	2AA	2AA	2AA	BP	2AA
	Concentrations (μg/plate)	3	3	3	40	3
	Mean No. of colonies/plate (average of 2 ± SD)	1218 ± 251	130 ± 12	382 ± 42	200 ± 48	718 ± 231

Plate incorporation

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 2 plates ± Standard deviation)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	TA102	TA98	TA97a
–	0	145 ± 17	12 ± 3.4	186 ± 3.6	8 ± 2.2	153 ± 27.3
–	37.5	158 ± 45	7 ± 3	136 ± 47	7 ± 3	122 ± 54
–	75	131 ± 19	8 ± 4	128 ± 42	13 ± 5	125 ± 20
–	150*	146 ± 7	6 ± 4	129 ± 28	11 ± 5	115 ± 37
Positive controls, –S9	Name	NaN3	NaN3	NPD	NPD	NPD
	Concentrations (μg/plate)	6	6	80	80	80
	Mean No. of colonies/plate (average of 2 ± SD)	1001 ± 0	1001 ± 0	1001 ± 0	1001 ± 0	1001 ± 0
+	0	153 ± 7	13 ± 1.7	137 ± 43.2	10 ± 3.4	141 ± 23.6
+	37.5	95 ± 16	8 ± 3	86 ± 25	10 ± 4	104 ± 31
+	75	104 ± 21	11 ± 3	117 ± 15	10 ± 4	156 ± 29
+	150*	120 ± 10	5 ± 2	134 ± 44	6 ± 2	89 ± 28
Positive controls, +S9	Name	2AA	2AA	2AA	BP	2AA
	Concentrations (μg/plate)	3	3	3	40	3
	Mean No. of colonies/plate (average of 2 ± SD)	1001 ± 0	1001 ± 0	1001 ± 0	105 ± 6	1001 ± 0

Preincubation

*Remark: 150 µg/plate was the highest concentration of test substance due to toxicity testing

NPD: 4-Nitro-1,2-phenylene diamine

NaN3: sodium azide

2AA: 2-Amino-anthracene

BaP: Benzo-a-pyrene

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative