Copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, [[3-(dimethylamino)propyl]amino]sulfonyl derivs.

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

For C.I. Direct Blue 264 no mutagenic or clastogenic effects were observed in a bacterial reverse mutation assay, in an in vitro gene mutation assay (HPRT) and in an in vitro micronucleus test.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1998

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Non-GLP

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

non GLP; content not given

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Target gene:

Histidine biosynthesis

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

Negative control
C.I. Direct Blue 264: 5000, 1581, 500, 158, 50, 16 µg/plate
Positive controls:
Na-azide: 10 µg/plate (TA1535)
Nitrofurantoin: 0.2 µg{plate (TA100)
4-nitro-1,2-phenylene diamine: 10 µg/plate (TA1537); 0.5 µg/plate (TA98)
Cumene: 50 µg/plate (TA102)
2-Aminoanthracene: 3 µg/plate

Due to precipitation of the substance, doses ranging from 100 to 3200µg/tube were chosen for the repeat tests.

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

cumene hydroperoxide

other: nitrofurantoin, 4-nitro-1,2-phenylene diamine, 2-aminoantracene

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

No bacteriotoxic effects up to 158 µg/plate; total bacterial count remained unchanged an no growth inhibition was seen.

At doses above 158 µg/plate strain-specific cytotoxicity.

Compound precipitation started at 500 µg/plate; and 5000 µg/plate could not be used for evaluation.

Conclusions:

Interpretation of results: negative

Executive summary:

No evidence of mutagenic activity in plate incorporation test and preincubation test in the presence and absence of S9 mix.

Reference 2

Endpoint:

in vitro cytogenicity / micronucleus study

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: OECD TG 487 In vitro mammalian cell micronucleus test

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian cell micronucleus test

Target gene:

Micronucleus test

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Metabolic activation system:

S9 mix; Aroclor 1254-induced male Sprague-Dawley rats.

Test concentrations with justification for top dose:

The 4 hours treatment was conducted with concentrations of 2.5, 5, 10, 25, 50, 100 and 200 µg/mL without and with S9 mix.

In the independent repeat test, the treatment time in the experiment without S9 mix was extended to 24 hours with concentrations of 1, 2.5, 5, 10, 25, 50 and 100 µg/mL.

Vehicle / solvent:

DMSO

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

cyclophosphamide

mitomycin C

other: vinblastine sulfate salt

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

The micronucleus test showed no biologically relevant increase in the frequencies of micronucleus containing V79 cells treated with C.I. Direct Blue 264 in the absence ( 4 hours or 24 hours treatment) or in the presence of S9 mix.

Conclusions:

Interpretation of results: negative

Executive summary:

Evaluation of the data does not indicate that C.I. Direct Blue 264 is a mutagen in the micronucleus test in vitro, when tested up to cytotoxic and precipitating concentrations in the absence or presence of metabolic activation.

Reference 3

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Principles of method if other than guideline:

The study was performed to investigate the potential of C.I. Direct Blue 264 to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster. The assay was performed in two independent experiments, using two parallel cultures each. The first main experiment was performed with and without liver microsomal activation and a treatment period of 4 hours. The second experiment was performed with a treatment time of 4 hours with and 24 hours without metabolic activation.

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Target gene:

HPRT locus in V79 cells

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

Experiment I
Exposure period: 4 hours (with and without S9 mix)
Concentrations without S9 mix: 125, 250, 500, 750 and 1000(p) µg/mL
Concentrations with S9 mix: 15.6, 31.3, 62.5(p), 125(p) and 250(p) µg/mL

Experiment II
Exposure period: 4 hours (with and without S9 mix)
Concentrations without S9 mix: 3.9, 7.8, 15.6 and 31 µg/mL
Concentrations with S9 mix: 15.6, 31.3, 62.5(p), 125(p) and 250(p) µg/mL

p = precipitation visible to the unaided eye

Vehicle / solvent:

DMSO

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

7,12-dimethylbenzanthracene

ethylmethanesulphonate

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

The study was performed to investigate the potential of C.I. Direct Blue 264 to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster.

The assay was performed in two independent experiments, using two parallel cultures each. The first main experiment was performed with and without liver microsomal activation (S9 mix) and a treatment period of 4 hours. The second experiment was performed with a treatment time of 4 hours with and 24 hours without metabolic activation.

The highest concentration of the pre-experiment (2000 μg/mL) was limited by the solubility properties of C.I. Direct Blue 264 in DMSO and aqueous medium. The maximum concentration of the main experiments was limited by cytotoxic effects and precipitation of the test item.

No substantial and reproducible dose dependent increase of the mutation frequency was observed in both main experiments.

Appropriate reference mutagens, used as positive controls, induced a distinct increase in mutant colonies and thus, showed the sensitivity of the test system and the activity of the metabolic activation system.

In conclusion it can be stated that under the experimental conditions reported C.I. Direct Blue 264 did not induce gene mutations at the HPRT locus in V79 cells. Therefore, C.I. Direct Blue 264 is considered to be non-mutagenic in this HPRT assay.

 

Conclusions:

Interpretation of results (migrated information):
negative

Executive summary:

The study was performed to investigate the potential of C.I. Direct Blue 264 to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster.

The assay was performed in two independent experiments, using two parallel cultures each. The first main experiment was performed with and without liver microsomal activation (S9 mix) and a treatment period of 4 hours. The second experiment was performed with a treatment time of 4 hours with and 24 hours without metabolic activation.

The highest concentration of the pre-experiment (2000 μg/mL) was limited by the solubility properties of C.I. Direct Blue 264 in DMSO and aqueous medium. The maximum concentration of the main experiments was limited by cytotoxic effects and precipitation of the test item.

No substantial and reproducible dose dependent increase of the mutation frequency was observed in both main experiments.

Appropriate reference mutagens, used as positive controls, induced a distinct increase in mutant colonies and thus, showed the sensitivity of the test system and the activity of the metabolic activation system.

In conclusion it can be stated that under the experimental conditions reported C.I. Direct Blue 264 did not induce gene mutations at the HPRT locus in V79 cells. Therefore, C.I. Direct Blue 264 is considered to be non-mutagenic in this HPRT assay.

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

No evidence of mutagenic activity was observed in a reliable bacterial reverse mutation assay using 5 bacterial strains (TA 1537, TA1535, TA102, TA100, TA98) similar to OECD TG 471, but not conducted according to GLP.

No evidence of mutagenic activity was reported in a gene mutation assay in Chinese Hamster V79 cells (HPRT) conducted according to OECD TG 474 and GLP.

No clastogenic or aneugenic activity was observed in an in vitro mammalian cell micronucleus test conducted according to OECD TG 487 and GLP.

Justification for selection of genetic toxicity endpoint
For C.I. Direct Blue 264 no mutagenic or clastogenic effects were observed in a bacterial reverse mutation assay, in an in vitro gene mutation assay (HPRT) and in an in vitro micronucleus test.

Justification for classification or non-classification

For C.I. Direct Blue 264 no mutagenic or clastogenic effects were observed in a bacterial reverse mutation assay, in an in vitro gene mutation assay (HPRT) and in an in vitro micronucleus test.

According to CLP classification criteria (Regulation (EC) No 1272/2008) a classification is not justified.