Disodium 4-hydroxy-3-[(4-sulphonatonaphthyl)azo]naphthalenesulphonate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The test material Carmoisine is not a mutagen in vitro.

Link to relevant study records

Reference

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:

other: Data is from peer reviewed publication

Principles of method if other than guideline:

The induction of mutations was studied in modified liquid fluctuation tests a histidine auxotroph of Salmonella typhimurium (specific for frameshifts) for the test compound Carmoisine.

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Species / strain / cell type:

S. typhimurium TA 1538

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Test concentrations with justification for top dose:

5 mg/mL

Vehicle / solvent:

Vehicle
Deionised water
- Justification for choice of solvent/vehicle: No data available

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: No data available
- Exposure duration: 72-96 hrs
- Expression time (cells in growth medium): 72-96 hrs
- 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: No data available

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: The food colour was made up in deionized water and membrane-sterilized prior to use. The test material was tested at its maximum sublethal concentration.

Evaluation criteria:

The test material was considered positive only if it resulted in significant more turbid tubes in a treated series when compared with an untreated set of tubes

Statistics:

Chi square test

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Remarks on result:

other: strain/cell type:

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

Carmoisine failed to induce gene mutation in the bacterial tester strain Salmonella typhimurium TA 1538 and hence is negative for gene mutation in vitro both in the presence and absence of metabolic activation system.

Executive summary:

Fluctuation test was performed for the test chemical Carmoisine used at a concentration of 5 mg/mL in liquid medium for 72-96 hrs. The tester strains used wereSalmonella typhimuriumTA 1538 both in thepresence and absence of metabolic activation system. The assay was performed for each bacterium in three separate experiments.

 

The given test material failed to induce genotoxicity in the bacteria Salmonella typhimuriumTA 1538 and hence is negative for gene mutation in vitro.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Gene mutation in vitro:

Prediction model based estimation and data from peer reviewed publications was used to determine the mutagenic nature of the test compound Carmoisine (CAS no 3567 -69 -9). The summary is as mentioned below:

Fluctuation test was performed (Haveland-Smith and Combes, 1979) for the test chemical Carmoisine used at a concentration of 5 mg/mL in liquid medium for 72-96 hrs. The tester strains used were Salmonella typhimurium TA 1538 and E.coli uvrA both in the presence and absence of metabolic activation system. The assay was performed for each bacterium in three separate experiments. The given test material failed to induce genotoxicity in the bacteria Salmonella typhimurium TA 1538 and hence is negative for gene mutation in vitro.

Carmoisine was used at a concentration of 5 mg/mL and its genotoxic effect was studied (Haveland-Smith and Combes, 1979) in the bacteria E. coli strains WP2 trp uvrA and WP100 trp uvrA recA by rec plate assay in the agar medium. The given test material is negative for genotoxicity in the bacteria E. coli strains WP2 trp uvrA and WP100 trp uvrA recA and hence is negative for gene mutation in vitro.

Bacterial gene mutation assay was performed by Cameron et al, 1979 for the test material C. I -ACID RED 14 in Salmonella typhimurium TA1535, TA1537, TA1538, TA100 and TA98 strains at a dose range of 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. The dye C. I -ACID RED 14 was found to be negative to induce gene mutation with and without S9 mix in Salmonella typhimurium strains TA1535, TA1537, TA1538, TA100 and TA98 by Plate-incorporation assay.

FMN modified assay was performed by Cameron et al, 1979 in the Salmonella typhimurium strains TA98 and TA100 for the test chemical C. I ACID RED 14. The strains were exposed to a concentration of 333.0, 1000.0, 3333.0, 6666.0 and 10000.0 µg/plate in the preincubation protocol. Positive control (Trypan blue) and solvent control were incorporated in the study. All plates were counted on an Artek automated counter, which was calibrated before use. The test material C. I ACID RED 14 failed to induce gene mutation in Salmonella typhimurium strains TA98 and TA100 in the FMN-modified assay.

L5178Y TK +/- mouse lymphoma assay was performed by Cameron et al, 1979 using C. I ACID RED 14 at different concentrations both with and without S9 metabolic activation system.Cells in duplicate cultures were exposed to the test chemical, positive control, and solvent control for 4 h at 37 ° C; washed twice with growth medium; and maintained at 37 °C for 48 h in log phase growth to allow recovery and mutant expression. The cultures were adjusted to 0.3 × 10⁶cells/ml at 24-h intervals. They were then cloned in soft agar medium containing Fischer's medium, 20% horse serum, 2 mM sodium pyruvate, 0.02% Pluronic F-68 and 0.35% Noble agar. Resistance to trifluorothymidine (TFT) was determined by adding 3 µg/ml TFT to one set of plates. The 100 X stock solution of TFT in saline was stored at -70°C and thawed immediately before use. Plates were incubated at 37°C in 5% CO 2 in air for 12 days, and then counted with an automatic colony counter. Mutant frequencies were expressed as mutants per 10⁴surviving cells. In general, a response was considered positive if there was a dose-related increase in the mutant frequency above the spontaneous control frequency, with a 2-fold increase at more than 1 dose and relative total growth greater than 10%. The C. I ACID RED 14 did not induce mutation and hence was negative (with and without S9 mix) in L5178Y TK +/- mouse lymphoma assay.

Based on prediction done using Danish QSAR database, it can be estimated that the substance Disodium 4-hydroxy-3-[(4-sulphonatonaphthyl)azo] naphthalenesulphonate is non genotoxic to S. typhimurium TA 1535, TA 1537, TA 98 and TA 100.

Based on the studies reviewed, the test material Carmoisine is not a mutagen.

Justification for selection of genetic toxicity endpoint
The test material Carmoisine failed to induce mutation in the Salmonella typhimurium TA 1538 and hence in not a mutagen.

Justification for classification or non-classification

Based on the key study and its supporting data, the test material is not a mutagen in vitro.