Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Peer reviewed journal research article

Data source

Reference
Reference Type:
publication
Title:
Salmonella Mutagenicity Tests: III. Results From the Testing of 255 Chemicals
Author:
Errol Zeiger, Beth Anderson, Steve Haworth, Timothy Lawlor, Kristien Mortelmans, and William Speck
Year:
1987
Bibliographic source:
Environmental Mutagenesis Volume 9, Supplement 9: 1 -110

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: Refer below principle
Principles of method if other than guideline:
The mutagenicity potential of the Rhodanine was studied by conducting experiment in Salmonella typhimurium strains TA100, TA1535, TA1537, TA98.
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: Rhodanine
- Molecular formula: C3H3NOS2
- Molecular weight: 133.1947 g/mol
- Substance type: Organic
Specific details on test material used for the study:
- Name of test material: Rhodanine
- Molecular formula: C3H3NOS2
- Molecular weight: 133.1947 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: 97%
- Impurities (identity and concentrations): 3%

Method

Target gene:
Histidine
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
No data
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
S9 fractions of Aroclor 1254-induced, male Sprague-Dawley rat and male Syrian hamster livers.
Test concentrations with justification for top dose:
0, 10, 33, 100, 333, 1000 µg/Plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Solubility of test substance in solvent
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 4-nitro-o-phenylenediamine: TA98 (Without metabolic activation). 2-Aminoanthracene: TA100, TA1535, TA1537, TA98 (With metabolic activation).
Details on test system and experimental conditions:
METHOD OF APPLICATION: Preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 2 days
- Expression time: 2 days
- Selection time: No data
- Fixation time: No data

SELECTION AGENT: No data
SPINDLE INHIBITOR: No data
STAIN: No data

NUMBER OF REPLICATIONS: 3

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data

- Other:

OTHER:
Rationale for test conditions:
No data
Evaluation criteria:
An individual trial was judged mutagenic (+) if a dose-related increase over the corresponding solvent control was seen, and it was judged weakly mutagenic (+W) if a low-level dose response was seen. A trial was considered questionable (?) if a dose related increase was judged insufficiently high to justify a call of " + W," if only a single dose was elevated over the control, or if a non-dose-related increase was seen.

The distinctions between a weak mutagenic response and a mutagenic response, or between a weak mutagenic response and a questionable mutagenic response are highly subjective.

A chemical was judged to be mutagenic (+), or weakly mutagenic (+W), if it produced a reproducible, dose-related increase in his+ revertants over the corresponding solvent controls in replicate trials. A chemical was considered to be questionable (?) if a reproducible increase of his+ revertants did not meet the criteria for either a " + " or " + W," or if only single doses produced an increase in his+ revertants in repeat trials. The chemicals were decoded by the chemical repository only after a determination had been made regarding their mutagenicity or no mutagenicity.

Statistics:
MEAN, SEM

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA100, TA1535, TA1537, TA98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
No data

Any other information on results incl. tables

Results for Rhodanine

Dose

                                 TA100                                 

NA

(-)

10% HLI (-)

10 % RLI (-)

µg/plate

MEAN

SEM

MEAN

SEM

MEAN

SEM

0

126

18.3

208

15.1

177

13.9

10

153

2.1

221

14.4

201

13

33

141

3.2

204

8

193

17.4

100

147

5.6

202

13.6

194

11.1

333

131

11.4

187

20.1

175

8.2

1000

103

5.7

168

12

168

11

POS

525

8.1

806

48.4

376

16.6

Dose

                                TA1535                     

NA

(-)

10% HLI (-)

10 % RLI (-)

µg/plate

MEAN

SEM

MEAN

SEM

MEAN

SEM

0

5

0.6

5

0.0

6

0.6

10

4

0.3

6

0.7

8

0.6

33

5

0.7

5

0.6

7

0.6

100

3

0.3

5

0.6

6

0.6

333

5

1.8

4

0.0

7

1.2

1000

5

2.1

3

0.3

4

0.7

POS

536

47.1

35

3.8

27

0.7

Dose

                                  TA98                       

NA

(-)

10% HLI (-)

10 % RLI (-)

µg/plate

MEAN

SEM

MEAN

SEM

MEAN

SEM

0

14

0.3

25

2.6

21

2.1

10

19

4.0

25

4.6

24

2.0

33

14

1.5

30

1.8

27

3.5

100

12

0.0

24

2.9

24

1.8

333

12

0.3

20

3.2

22

4.5

1000

10

1.2

18

1.8

15

3.2

POS

411

32.2

650

34.0

244

9.6

Abbreviations:                     

NA: Not activated

RLI: Aroclor 1254-induced rat liver S-9;

HLI: Aroclor 1254-induced hamster liver

POS: Positive control

Applicant's summary and conclusion

Conclusions:
The mutagenic potential of Rhodanine was evaluated by performing experiment in Salmonella typhimurium strains TA100, TA1535, TA1537, TA98 with and without metabolic activation, test chemical not induce mutation in bacterial tester strains. Therefore, Rhodanine was considered to be not mutagenic In vitro.
Executive summary:

The mutagenicity effect of Rhodanine was evaluated by performing mutagenic assay in Salmonella typhimurium. Salmonella typhimurium strainsTA100, TA1535, TA1537, TA98 involved in the study. Mutagenic assay was performed by preincubation method with and without metabolic activation i.e. S9 fractions of Aroclor 1254 -induced, male Sprague-Dawley rat and male Syrian hamster livers. Test chemical in concentrations 0, 10, 33, 100, 333, 1000 µg/Plate were tested. The test chemical, Salmonella culture, and S9 mix were incubated at 37°C without shaking for 20 min. The top agar was added, and the contents of the tubes were mixed and poured onto the surface of petri dishes that contained Vogel-Bonner medium [Vogel and Bonner. The histidine-revertant (his+) colonies arising on these plates were counted following 2 days incubation at 37°C. The plates were hand counted when precipitated was present, otherwise automatic colony counters were used. All doses of test chemical were tested in triplicate. Concurrent solvent and positive controls were run with each trials. The positive control substance without metabolic activation were Sodium azide (TA100, TA1535), 9-aminoacridine (TA1537) and 4-nitro-o-phenylenediamine (TA98). Positive control for metabolic activation was 2-aminoanthracene for all strains.

The number of revertants for test chemical with and without metabolic activation not more than solvent control, hence test chemical was not induce mutation. Therefore Rhodanine was considered to be not mutagenic In vitro.