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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
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:
In Vitro Genotoxicity of Dyes Present in Colored Smoke Munitions
Author:
A.L. Brooks, F.A. Seiler, R.L. Hanson, and R.F. Henderson
Year:
1989
Bibliographic source:
Environmental and Molecular Mutagenesis 13:304-313 (1989)

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:
Gene mutation toxicity study was performed to determine the mutagenic nature of Solvent red 1
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: Solvent Red 1
- Molecular formula: C17H14N2O2
- Molecular weight: 278.31 g/mol
- Substance type: Organic
Specific details on test material used for the study:
- Name of test material: Solvent red 1 / 1 -((2Methoxyphenyl) azo-2-naphthol)
- Molecular formula: C17H14N2O2
- Molecular weight: 278.31 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: 99%
- Impurities (identity and concentrations): 1%

Method

Target gene:
Histidine
Species / strain
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell lines (if applicable):
Not applicable
Additional strain characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
Aroclor induced rat liver microsomes (S-9)
Test concentrations with justification for top dose:
Six concentration between 0-800 µg/plate
Vehicle:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The chemical was soluble in DMSO
Controls
Negative controls:
not specified
Solvent controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
benzo(a)pyrene
other: 2-aminoanthracene (2-AA)
Details on test system and conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

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

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

NUMBER OF REPLICATIONS: Triplicate

NUMBER OF CELLS EVALUATED: No data

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

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

OTHER: No data
Rationale for test conditions:
No data
Evaluation criteria:
The plates were observed for a doubling of revertants/plate
Statistics:
The average revertant frequency per plate was plotted against the concentration of the dyes and concentration response relationship determined for each dye for each S-9 concentration over the linear portion of the curve. To test the null hypothesis that the slope was not different from
zero, the data were first fit to a straight line by a least squares method, then the derived slope was tested using a Z-test with a linear dose axis and a
significance level of 0.05.

Results and discussion

Test results
Species / strain:
other: TA98, TA1538, TA100, and TA1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not specified
Positive controls valid:
yes
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data
- Effects of osmolality: No data
- Evaporation from medium: No data
- Water solubility: No data
- Precipitation: No precipitate was observed
- Other confounding effects: No data

RANGE-FINDING/SCREENING STUDIES: No data

COMPARISON WITH HISTORICAL CONTROL DATA: No data

Any other information on results incl. tables

Concentration (µg/plate)

Strain

S9 (30µL/plate)

Background (X±SE)

Highest response (X±SE)

Slope±SE (revertants/µg)

0-800

TA98

-

30±3

32±1

0.002±0.003

 

+

36±3

43±2

0.008±0.003

TA1538

-

13±2

23±2

0.003±C 0.004

 

+

21±2

29±4

-0.001±0.005

TA100

-

114±3

134±6

0.010±0.006

 

+

121±5

170±10

0.01±0.02

TA1535

-

21±3

32±3

-0.004±0.006

 

+

10±1

15±5

0.002±0.002

Applicant's summary and conclusion

Conclusions:
Solvent red 1 did not induce mutation in the range finding study conducted using Salmonella typhimurium strains TA98, TA1538, TA100, and TA1535 both in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
Executive summary:

Gene mutation toxicity study was performed to determine the mutagenic nature of Solvent Red 1. The study was conducted as a part of range finding study using Salmonella typhimurium strains TA98, TA1538, TA100, and TA1535 with and without S9 metabolic activation system. The test chemical was dissolved in DMSO and used at six concentration from 0-800µg/plate. Three plates were used per concentration and the plates were observed for an increase in mutation frequency twice that observed in the controls.Solvent red 1 did not induce mutation in the reange finding study conducted usingSalmonella typhimurium strains TA98, TA1538, TA100, and TA1535 both in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.