Reaction mass of stearoyl sarcosine and palmitoyl sarcosine

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:

17 Oct - 07 Nov 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted on 1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted in 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht, Mainz, Germany

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N-(1-oxooctadecyl)sarcosine

EC Number:

205-539-7

EC Name:

N-(1-oxooctadecyl)sarcosine

Cas Number:

142-48-3

Molecular formula:

C21H41NO3

IUPAC Name:

N-methyl-N-stearoylglycine

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

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

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, +S9, -S9: 53, 157, 508, 1507, and 5005 µg/plate
Second experiment, +S9, -S9: 323, 635, 1300, 2607, and 4792 µg/plate

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

benzo(a)pyrene

other: 4-Nitro-1,2-phenylene diamine (NPD); 2-Amino-anthracene (2-AA)

Remarks:

NPD; 20 µg/plate in water, -S9, TA98, TA 97a and TA 102; sodium azide (1 µg/plate in water, -S9, TA1535 and TA100); 2-AA; 1 µg/plate in DMSO, +S9, all strains; benzo-a-pyrene (BaP; 20 µg/plate in DMSO, +S9, all strains)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation) and preincubation
DURATION
- Preincubation period: 48 h
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: four in two different experiments

DETERMINATION OF CYTOTOXICITY
- Method: other: Four plates per strain were incubated with the corresponding dose of the test item on maximal soft agar, to test the toxicity of the following concentration: 5005 µg/plate.
The titre was determined according to the following criterion: the determination of the titre should give a number of at least 10E9 cells/mL, correlating to 100 colonies/plate after dilution.
The test item was considered non-toxic, if the quotient titre/tox is below 2.

Evaluation criteria:

A test item is considered to have mutagenic potential, if a significant, reproducible increase of revertant colonies per plate (increase factor ≥ 2) in at least one strain can be observed.
A concentration-related increase over the range tested can also be taken as a sign of mutagenic activity.

Statistics:

Mean and standard deviation

Results and discussion

Test results

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:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: Concentrations of the test item are stated as nominal concentrations, as suspensions had to be tested due to the poor solubility of the test item. As no complete dissolution was possiblem undissolved particles were visible on the plates.

Any other information on results incl. tables

First Experiment

Confirmation of the criteria and validity

The treatments for the sterility control and the determination of the titre did not show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range of the test laboratory (historical data of the laboratory). All positive controls showed mutagenic effects with and without metabolic activation.

Solubility and Toxicity

The test item was suspended in deionised water. All concentrations were weight directly. As no complete dissolution was possible, undissolved particles were visible on the plates.

No signs of toxicity towards the tested strains could be observed. The background lawn was visible and the number of revertant colonies was not reduced.

Mutagenicity

No significant increase of the number of revertant colonies in the treatments with and without metabolic activation could be observed. No concentration-related increase over the tested range was found.

Therefore the test item is stated as not mutagenic under the test conditions.

Second Experiment

Confirmation of the criteria and validity

The treatments for the sterility control and the determination of the titre did not show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range of the test laboratory. All positive controls showed mutagenic effects with and without metabolic activation.

Solubility and toxicity

The test item was suspended in deionised water. All concentrations were weights directly. As no complete dissolution was possible, undissolved particles were visible on the plates. No signs of toxicity towards the tested strains could be observed. The background lawn was visible and the number of revertant colonies was not significantly reduced.

Mutagenicity

No significant increase of the number of revertant colonies in the treatments with and without metabolic activation was observed. No concentration-related increase over the tested range was found.

Therefore, the test item is stated as not mutagenic under the test conditions.

Table 1 First experiment: mean revertants

Strain	TA97a	TA97a	TA 98	TA 98	TA100	TA100	TA102	TA102	TA1535	TA1535
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
H2O	111±7.6	120±4.2	18±2.5	15±1.2	103±7.1	98±9.6	134±5.2	143±9.2	19±3.4	20±2.9
DMSO	112±8.2	105±5.2	18±2.2	16±2.2	94±15.6	87±9.5	134±8.3	132±7.4	18±1.9	22±1.3
Positive controls*	611±72	726±29	199±20	203±4	441±43	437±26	509±27	543±22	220±14	232±17
5005µg/plate	107±8	116±2	16±2	20±1	83±2	80±5	142±12	152±3	18±3	19±3
1507µg/plate	144±7	119±9	16±2	17±2	85±2	89±6	137±14	144±9	16±3	17±1
508 µg/plate	106±10	109±6	15±2	16±1	89±3	93±12	143±9	134±6	17±3	17±2
157 µg/plate	112±2	111±8	15±2	18±2	85±5	83±12	152±3	137±14	20±4	17±2
53 µg/plate	109±5	107±6	16±2	18±3	81±3	78±5	152±10	129±8	14±2	16±2

 

 Table 2 Second experiment: mean revertants

Strain	TA97a	TA97a	TA 98	TA 98	TA100	TA100	TA102	TA102	TA1535	TA1535
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
H2O	117±5	103±3.3	17±2.6	17±2.6	110±3.8	105±3.6	154±7.1	144±5.8	21±2.2	21±3.4
DMSO	116±4.6	113±4.9	16±0.6	15±1.4	97±8	104±3	156±4.8	147±8.5	16±3.5	23±3.0
Positive controls*	629±38	570±25	206±14	235±16	538±58	550±33	628±51	598±15	513±39	474±11
4792µg/plate	115±5	109±4	16±2	13±2	84±4	85±5	151±6	138±3	19±5	12±1
2607µg/plate	115±4	114±3	18±2	16±2	81±6	90±2	132±13	127±11	16±4	14±2
1300 µg/plate	105±5	104±3	17±3	17±2	92±6	89±3	141±3	146±9	16±4	17±5
635 µg/plate	107±5	112±3	14±3	17±1	81±8	108±4	146±8	145±5	14±3	17±3
323 µg/plate	117±4	118±4	14±3	12±1	75±3	82±6	134±12	143±6	19±2	15±2

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative