L-Proline, 4-(phenylthio)-, (4S)-

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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1997

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(2S,4S)-4-(phenylsulfanyl)pyrrolidine-2-carboxylic acid

EC Number:

617-252-8

Cas Number:

81653-77-2

Molecular formula:

C11H13NO2S

IUPAC Name:

(2S,4S)-4-(phenylsulfanyl)pyrrolidine-2-carboxylic acid

Method

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 102

Metabolic activation:

with and without

Metabolic activation system:

Mammalian Microsomal Fraction S9 mix

Test concentrations with justification for top dose:

The test item concentrations to be applied in thè main experiments were chosen according to the results of the pre-experiment.
5000 mcg/plate was selected as the maximum concentration. The concentration range covered two logarithmic decades. Two independent experiments were performed with the following concentrations:
31.6, 100, 316, 1000, 2500 and 5000 mcg/plate

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

solvent control

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

methylmethanesulfonate

other: 4-nitro-O-phenylene-diamine; 2-amino-anthracene

Details on test system and experimental conditions:

Positive Controls
- Without metabolic activation
Tester Strains: S. typhimurium: TA 100, TA 1535
Name: sodium azide
Lot No.: K28585088
Purity: 99%
Dissolved in: aqua dest.
Concentration: 10 mcg/plate

Tester Strains: S. typhimurium TA 98, TA 1537
Name: 4-nitro-o-phenylene-diamine, 4-NOPD
Lot No.: 416325
Purity: 97%
Dissolved in: DMSO
Concentration: 10 mcg/plate for TA 98, 40 mcg/plate for TA 1537

Tester Strains: S. typhimurium TA 102
Name: Methyl methane sulfonate, MMS
Lot No.: 126K3721
Purity: 99%
Dissolved in: Aqua dest.
Concentration: 1 mcL/plate

-With metabolic activation
Tester Strains: S. typhimurium'. TA 98, TA 100, TA 1535, TA 1537 and TA 102
Name: 2-aminoanthracene, 2-A A
Lot No.: SI 1804-114
Purity: 96%
Dissolved in: DMSO
Concentration: 2.5 mcg/plate; 10 mcg/plate for TA 102

The test item was investigateci for its potential to induce gene mutations according to the plate incorporation test (experiment I) and the pre-incubation test (experiment II), after a pre-test to verify the toxicity.
The concentrations, including the Controls, were tested in triplicate.

Results and discussion

Test results

Test results 1

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

Test results 2

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Additional information on results:

No precipitation of the test item was observed in any of the tester strains used in experiment I and II (with and without metabolic activation).
No toxic effects of the test item were noted in any of the five tester strains used up to the highest dose group evaluated (with and without metabolic
activation) in experiment I and II.

Applicant's summary and conclusion

Conclusions:

The test item is considered to be non-mutagenic in this bacterial reverse mutation assay.

Executive summary:

In order to investigate the potential of the test item for its ability to induce gene mutations the plate incorporation test (experiment I) and the pre-incubation test (experiment II) were performed with the Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 102. Each assay was conducted with and without metabolic activation. The concentrations, including the Controls, were tested in triplicate. The following concentrations of the test item were prepared and used in the experiments:

31.6, 100, 316, 1000, 2500 and 5000 mcg/plate.

No biologically relevant increases in revertant colony numbers of any of the five tester strains were observed at any concentration level, neither in the presence nor absence of metabolic activation in experiment I and II.

Therefore, the test item is considered to be non-mutagenic in this bacterial reverse mutation assay.