N-(1-oxooctyl)glycine

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:

august to september 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: complete study according to OECD 471 and GLP guidelines

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

not applicable

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N-(1-oxooctyl)glycine

EC Number:

238-122-3

EC Name:

N-(1-oxooctyl)glycine

Cas Number:

14246-53-8

Molecular formula:

C10H19NO3

IUPAC Name:

N-(1-oxooctyl)glycine

Constituent 2

Reference substance name:

LIPACIDE C8G

IUPAC Name:

LIPACIDE C8G

Details on test material:

- Name of test material (as cited in study report): LIPACID C8G
- Substance type: mono constituant substance
- Physical state: solid powder
- Analytical purity: 99%
- Impurities (identity and concentrations): <1%
- Lot/batch No.: 99056001
- Storage condition of test material: under room temperature

Method

Target gene:

mutation in histidine biosynthesis: His D3052 (frameshift), His G46 (base pair-substitution and his C 3076 frameshift

Species / strain

Species / strain 1

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not specified

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

0; 0,15; 0,5; 1,5; 5; 15; 50; 150; 500; 1500; 5000 µg/plate

Vehicle / solvent:

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

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Remarks:

for TA100 ; TA1535 and WP2uvrA- without S9

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

for TA1537 without S9

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

for TA98 without S9

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

for TA98 with S9 mix

Controls 5

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

Remarks:

For TA100; TA1537 ; TA1535 and WP2uvrA- with S9mix

Details on test system and experimental conditions:

1. preliminary toxicity study
the dose range of the test material was 0;1.15;0.5;1.5;5;15;50;150;500;1500;5000µg/plate. the study was performed by mixing 0.1ml of bacterial cultures (TA100 or WPuvrA-), 0.1ml of the test material formulation, 0.5ml of S9 mix or phosphate buffer and 2ml of molten, trace histidine or tryptophane supplemented , top agar and overlaying onto sterile plates of vogel-Bonner Minimal agar.10 concentrations of the test material and a vehiculme DMSo were tested . after 48 h of incubation at 37°C the plates were assessed for numbers of revertant colonies using a domino counter colony.

2.range finding study
3. main study

Evaluation criteria:

the test material may be considered to be positive in this test system if the following criteria are met: the test material should have induced a reproducible , dose related and statistically (dunnets method of linear regression (5)) significant increase in the revertant count in at least one strain of bacteria.

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

the test material was considered to be non mutagenic under the conditions of this study.

Executive summary:

1. Samonelle typhimurium strains TA1535, TA1537, TA98, and TA100 and Escherichia coli strain WP2uvrA- were treated with the test material using the Ames plate incorporation method at five dose levels, in triplicate, both with and without the addition of a rat liver homogenate metabolising system (10% liver S9 in standard co-factors). This method conforms to the guidelines for bacterial mutagenicity testing published by the major japanese Regulatory Authorities includi ng MITI, MHW, MOL and MAFF. It also meets the requirements of the OECD guidelines for testing of chemicals No.471. "Reverse mutation study", methodB14 of commission directive 92/69/EEC and the USA, EPA (TSCA) OPPTS harmonised guidelines. the dose range was determined in a preliminary toxicity assay and was 50 to 5000µg/plate in the range-finding study. The experiment was repeated on a separateday using the same dose range as the range-finding study, fresh cultures of the bacterial strains and fresh test material formulations.

2. The vehicule (Dimethyl sulphoxide) control plates gave counts of relevant colonies within the normal range.

3. Alle the positive control chemicals used in the test induced marked increases in the frequency of rlevant colonies, both with and without metabolic activation. thus the sensitivity of the assay and the efficacy of the S9 mix were validated.

4. The test material caused sporadic toxicity to several of the tester strains either as a weakened bacterial background lawn or a decrease in revertant colony frequency . the test material was, therefore, tested up to the maximum recommended up to the maximum recommended dose tested in either the presence or absence of S9 -mix.

5. No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test material, either with or without matabolic activation. the test material was considered to be non mutagenic under the conditions of this study.