9-(2-carboxyphenyl)-3,6-bis(diethylamino)xanthylium acetate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames test: The test chemical did not induce gene mutation in Salmonella typhimurium TA98, TA100, TA1538 in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data from various test chemicals

Justification for type of information:

Data is summarized based on the available information from various read across test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on 2 gene mutation studies as - WoE 2 and WoE 3
Gene mutation test was carried out to study the effects of the test chemical.

GLP compliance:

not specified

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium, other: TA 100 and TA 1538

Remarks:

2

Species / strain / cell type:

S. typhimurium, other: TA 100, TA 1535, TA 97, TA 98

Remarks:

3

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

2.S9 protein was obtained from the liver of Aroclor-induced male Sprague-Dawley rats.
3.The S-9 fractions of Aroclor 1254-induced, male Sprague-Dawley rat and male Syrian hamster livers were prepared

Test concentrations with justification for top dose:

2.100µg/plate or 1 mg/plate
3. 0, 33, 100, 333, 1000, 2000, 3333, 5000 or 10000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- 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

Positive control substance:

not specified

Remarks:

2

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

other: 2-Aminoanthracene (All strains + S9); 4-Nitro-o-phenylenediamine (TA98; -S9)

Remarks:

3

Details on test system and experimental conditions:

2.METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: No data available
- Exposure duration: 2 days (48 hrs)
- Expression time (cells in growth medium): 2 days (48 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: Duplicate

NUMBER OF CELLS EVALUATED: No data available

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

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

OTHER: No data available

3.METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 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: 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:

OTHER: No data available

Rationale for test conditions:

No data

Evaluation criteria:

2.A sample was judged mutagenic if it produced greater than twice the spontaneous background colonies at more than one dose or at the highest dose tested.
3.An individual trial was judged mutagenic (+) if a dose-related increase over the corresponding solvent control was seen, and it was judged weakly mutagenic C+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 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.

It 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.

Statistics:

2.To estimate mutagenic potency (revertant /µg) from linear dose-response curves, the method of Moore and Felton was used
3.No data

Species / strain:

S. typhimurium, other: TA 100 and TA 1538

Remarks:

2

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

Species / strain:

S. typhimurium, other: TA100, TA1535, TA97, TA98

Remarks:

3

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:

2.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: Spot test was performed with and without activation using the Salmonella typhimurium strains TA1538, TA98 and TA100. The spot test results were inconclusive. The dyes were then screened at 0.1 mg/plate and 1 mg/plate.

COMPARISON WITH HISTORICAL CONTROL DATA: No data available

ADDITIONAL INFORMATION ON CYTOTOXICITY: No data available
3.No data

Remarks on result:

other: No mutagenic potential observed

Conclusions:

The test chemical does not induce gene toxicity in the Salmonella typhimurium strains in the presence and absence of metabolic activation and hence it is negative for gene mutation.

Executive summary:

Data available for the test chemical was reviewed to determine the mutagenic nature of test chemical, The studies are as mentioned below:

Gene mutation study was performed to evaluate the mutagenic nature of test chemical using Salmonella typhimurium strain TA1538 and TA100. Bacteria were grown overnight in Oxoid nutrient broth, then refrigerated at 4-5^OC for a few hours before use. 0.1 ml of bacterial culture was added to 2 ml of 45°C molten top agar containing 0.01 mg histidine HCI and 0.012 mg biotin/ml, followed by the test sample in ≤0.2 ml DMSO. Finally, 0.5 ml of sodium phosphate buffer, pH 7.4 (no activation), or 0.5 ml of Aroclor-induced rat S9 mixture was added, and the mixture was poured on minimal glucose agar plates. Histidine revertant colonies were counted on a Biotran II automated colony counter after 2-day incubation at 37°C. 100µg/plate or 1 mg/plate of test chemical was used in the study. A sample was judged mutagenic if it produced greater than twice the spontaneous background colonies at more than one dose or at the highest dose tested. In the above mentioned study, test chemical failed to induce gene mutation in theSalmonella typhimuriumstrains TA1538 and TA100 with and without metabolic activation. Hence, the test chemical, is not likely to be a gene mutant in vitro.

In another study, Gene mutation toxicity study was performed to determine the mutagenic nature of test chemical using Salmonella typhimurium strains TA100, TA1535, TA97and TA98. Salmonella/microsome test was performed both in the absence and presence of liver S-9 obtained from Aroclor-induced male Sprague-Dawley rats and Syrian hamsters. Preincubation assay was performed at dose levels of 0, 33, 100, 333, 1000, 2000, 3333, 5000, 10000 µg/plate. The plates were preincubated for 20 mins and incubated futher for 48 hrs chemical exposure. Concurrent solvent and positive controls were also included in the study. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA98, TA100, TA1535 and TA 97 in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

 

Based on the observations made, the test chemical does not exibit gene mutation in vitro . Hence it is not likely to classify as a gene mutant in vitro as per the criteria mentioned in CLP regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Ames test:

Data available for the test chemical was reviewed to determine the mutagenic nature of test chemical, The studies are as mentioned below:

Gene mutation study was performed to evaluate the mutagenic nature of test chemical using Salmonella typhimurium strain TA1538 and TA100. Bacteria were grown overnight in Oxoid nutrient broth, then refrigerated at 4-5^OC for a few hours before use. 0.1 ml of bacterial culture was added to 2 ml of 45°C molten top agar containing 0.01 mg histidine HCI and 0.012 mg biotin/ml, followed by the test sample in ≤0.2 ml DMSO. Finally, 0.5 ml of sodium phosphate buffer, pH 7.4 (no activation), or 0.5 ml of Aroclor-induced rat S9 mixture was added, and the mixture was poured on minimal glucose agar plates. Histidine revertant colonies were counted on a Biotran II automated colony counter after 2-day incubation at 37°C. 100µg/plate or 1 mg/plate of test chemical was used in the study. A sample was judged mutagenic if it produced greater than twice the spontaneous background colonies at more than one dose or at the highest dose tested. In the above mentioned study, test chemical failed to induce gene mutation in theSalmonella typhimuriumstrains TA1538 and TA100 with and without metabolic activation. Hence, the test chemical, is not likely to be a gene mutant in vitro.

In another study, Gene mutation toxicity study was performed to determine the mutagenic nature of test chemical using Salmonella typhimurium strains TA100, TA1535, TA97and TA98. Salmonella/microsome test was performed both in the absence and presence of liver S-9 obtained from Aroclor-induced male Sprague-Dawley rats and Syrian hamsters. Preincubation assay was performed at dose levels of 0, 33, 100, 333, 1000, 2000, 3333, 5000, 10000 µg/plate. The plates were preincubated for 20 mins and incubated futher for 48 hrs chemical exposure. Concurrent solvent and positive controls were also included in the study. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA98, TA100, TA1535 and TA 97 in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

 

Based on the observations made, the test chemical does not exibit gene mutation in vitro . Hence it is not likely to classify as a gene mutant in vitro as per the criteria mentioned in CLP regulation.

Justification for classification or non-classification

Based on the observations made, the test chemical does not exibit gene mutation in vitro. Hence it is not likely to classify as a gene mutant in vitro as per the criteria mentioned in CLP regulation.