Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
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:
Data is from peer reviewed publication

Data source

Reference
Reference Type:
publication
Title:
Mutagenic activity of 27 dyes and related chemicals in the Salmonella/microsome and mouse lymphoma TK +/- assays
Author:
T.P. Cameron, T.J. Hughes , P.E. Kirby , V.A. Fung and V.C. Dunkel
Year:
1987
Bibliographic source:
Mutation Research, 189 (1987) 223-261

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
Mutagenicity testing was performed by the standard plate-incorporation assay for C I -ACID RED 14 (Carmoisine) in Salmonella typhimurium by using TA1535, TA1537, TA1538, TA98 and TA100 strains.
GLP compliance:
not specified
Type of assay:
bacterial gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: Carmoisine
- Molecular formula: C20H14N2O7S2.2Na
- Molecular weight: 502.4338 g/mol
- Substance type: organic
- Physical state: No data available
- Purity 85% dye
- Impurities (identity and concentrations):
15.0 %NaCI
>0 1% subsidiary colors
Specific details on test material used for the study:
- Name of test material: Carmoisine
- IUPAC name: disodium 4-hydroxy-3-[(4-sulfonato-1-naphthyl)diazenyl]naphthalene-1-sulfonate
- Molecular formula: C20H14N2O7S2.2Na
- Molecular weight: 502.4338 g/mol
- Substance type: organic
- Physical state: No data available
- Purity 85% dye
- Impurities (identity and concentrations): 15.0 %NaCI >0 1% subsidiary colors

Method

Target gene:
Histidine
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA1535, TA1537, TA1538, TA100 and TA98 were used.
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
S9 mix was prepared from Aroclor 1254-induced male Fischer 344 rats and Syrian golden hamsters.
Test concentrations with justification for top dose:
0, 333.0, 1000.0, 3333.0, 6666.0, 10000.0 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: The solvents used were water, dimethyl sulfoxide, and acetone (exact solvent details are not available)
- Justification for choice of solvent/vehicle: Test chemical solubility in the solvents mentioned
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
other: 2-aminoanthracene (All strains ; with S9)
Details on test system and experimental conditions:
METHOD OF APPLICATION: plate incorporation.

DURATION
- Preincubation period: No data available
- Exposure duration: No data available
- Expression time (cells in growth medium): No data available
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: The chemical was tested at 5 dose levels in triplicate.

NUMBER OF CELLS EVALUATED: No data available

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

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

OTHER: No data available







Rationale for test conditions:
No data available
Evaluation criteria:
A response was considered positive if there was a dose-related increase in the number of revertants above spontaneous solvent controls with a 2-fold increase for strains TA1535, TA1538, TA98 and TA100, and a 3-fold increase for TA1537
Statistics:
No data available

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA1535, TA1537, TA1538, TA100 and 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:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data available
- Effects of osmolality: No data available
- Evaporation from medium: No data available
- Water solubility: No data available
- Precipitation: No data available
- Other confounding effects: No data available

RANGE-FINDING/SCREENING STUDIES: The range of concentrations for testing was based on preliminary toxicity tests in which the viability of the bacterial cells on complete medium was measured at concentrations up to 10 mg/plate or to the limit of solubility. When solubility and toxicity were not limiting factors, the maximum concentration tested was 10 mg/plate.

COMPARISON WITH HISTORICAL CONTROL DATA: No data available

ADDITIONAL INFORMATION ON CYTOTOXICITY: No data available
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table: Mutagenicity of C. I. Acid red 14 in Salmonella Assay

Dose (µg/plate)

Revertants/plate

TA1535

TA1537

TA1538

(-)S9

(R)S9

(H)S9

(-)S9

(R)S9

(H)S9

(-)S9

(R)S9

(H)S9

DMSO

43±6

30±4

28±2

7±5

11±2

10±3

11±3

23±7

20±6

Positive control

484±22

260±18

183±11

453±117

48±15

124±13

1042±47

155±2

793±18

333

38±3

31±6

30±3

10±1

9±3

12±2

10±2

24±4

26±1

1000

36±12

26±6

31±1

6±3

11±4

13±2

11±3

24±7

26±6

3333

31±8

24±3

18±6

7±2

6±1

12±4

10±4

15±4

16±1

6666

23±3

14±2

18±3

7±2

9±2

10±3

14±5

11±3

18±2

10000

29±5

20±11

17±5

9±1

6±2

9±1

13±1

16±2

23±3

 

Dose (µg/plate)

Revertants/plate

TA98

TA100

(-)S9

(R)S9

(H)S9

(-)S9

(R)S9

(H)S9

DMSO

13±1

25±4

27±6

151±7

149±16

177±17

Positive control

423±32

1745±72

1848±122

574±24

1079±108

1446±55

333

25±3

32±3

30±4

157±6

175±1

175±16

1000

18±1

33±10

38±3

152±12

183±5

176±11

3333

22±5

19±1

28±4

137±10

142±27

164±5

6666

25±4

27±8

28±10

150±21

125±9

133±7

10000

21±5

22±5

22±6

158±19

134±17

144± 7

Applicant's summary and conclusion

Conclusions:
The dye C. I . Acid Red 14 did not induce gene mutation in Salmonella typhimurium strains TA1535, TA1537, TA1538, TA100 and TA98 in the presence and absence of S9 metabolic activation system by plate-incorporation assay and hence is not likely to classify as a gene mutant in vitro.
Executive summary:

Bacterial gene mutation assay was performed for the test material C. I Acid Red 14 (Carmoisine) in Salmonella typhimurium TA1535, TA1537, TA1538, TA100 and TA98 strains at a dose range of 0, 333.0, 1000.0, 3333.0, 6666.0 and 10000.0 µg/plate by Plate-incorporation method. Chemical was tested without metabolic activation and with S9 mix from Aroclor 1254-induced male Fischer 344 rats and Syrian golden hamsters. Appropriate positive and solvent controls were also incorporated in the study. Doses for the main study were based on the prelimicary study conducted. A response was considered positive if there was a dose-related increase in the number of revertants above spontaneous solvent controls with a 2-fold increase for strains TA1535, TA1538, TA98 and TA100, and a 3-fold increase for TA1537. The dye C. I . Acid Red 14 did not induce gene mutation in Salmonella typhimurium strains TA1535, TA1537, TA1538, TA100 and TA98 in the presence and absence of S9 metabolic activation system by plate-incorporation assay and hence is not likely to classify as a gene mutant in vitro.