[N,N,N',N',N'',N''-hexaethyl-29H,31H-phthalocyaninetrimethylaminato(2-)-N29,N30,N31,N32]copper tris(dodecylbenzenesulphonate)

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-01-24 to 2012-01-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP and OECD guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

[N,N,N',N',N'',N''-hexaethyl-29H,31H-phthalocyaninetrimethylaminato(2-)-N29,N30,N31,N32]copper tris(dodecylbenzenesulphonate)

EC Number:

278-150-3

EC Name:

[N,N,N',N',N'',N''-hexaethyl-29H,31H-phthalocyaninetrimethylaminato(2-)-N29,N30,N31,N32]copper tris(dodecylbenzenesulphonate)

Cas Number:

75247-18-6

Molecular formula:

C47 H49 Cu N11 .3 C18 H30 O3 S

IUPAC Name:

Copper,N,N,N',N',N'',N''-hexaethyl-29H,31H-phthalocyanine-C,C,C-trimethanaminato(2-)-κN29,κN30,κN31,κN32]-,dodecylbenzenesulfonate (1:3)

Details on test material:

Please refer to confidential details on test material

Method

Target gene:

Salmonella typhimurium: His (-/-)
Escherchia coli: Tryp (-/-)

Species / strain

Species / strain / cell type:

other: S.typhimurium TA 1535, TA 1537, TA 98, TA 100 and E.coli WP2 uvrA

Metabolic activation:

with and without

Metabolic activation system:

Aroclor-induced rat liver S9-mix

Test concentrations with justification for top dose:

Standard plate test: 0, 33, 100, 333, 1000, 2500 and 5000 µg/plate (with and without metabolic activation)
Preincubation test: 0, 33, 100, 333, 1000, 2500 and 5000 µg/plate (with and without metabolic activation)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Due to the limited solubility of the test substance in water, DMSO was used as vehicle, which
had been demonstrated to be suitable in bacterial reverse mutation tests and for which historical control data are available

Controls

Untreated negative controls:

yes

Remarks:

(sterillity control)

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene (+ S9); N-methyl-N'-nitro-N-nitrosoguanidine, 4-Nitro-o-phenylenediamine (NOPD), 9-Aminoacridine (AAC), 4-Nitroquinoline-N-oxide (4-NQO) (without S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: standard plate test (plate incorporation method) and preincubation

DURATION
- Incubation period: about 48-72 h at 37 °C in darkness
- Preincubation period: ca. 20 min

NUMBER OF REPLICATIONS:
- 3 test plates per dose or per control (2 independent experiments)


DETERMINATION OF CYTOTOXICITY
- decrease in the number of revertants
- clearing or diminution of the background lawn
- reduction in the titer
(for all test groups with and without S9-mix)

Titer:
The titer is generally determined only in the experimental parts with S9 mix both for the negative controls (vehicle only) and for the two highest doses in all experiments.

Evaluation criteria:

Acceptance criteria
Generally, the experiment is considered valid if the following criteria are met:
• The number of revertant colonies in the negative controls was within the range of the
historical negative control data for each tester strain.
• The sterility controls revealed no indication of bacterial contamination.
• The positive control substances both with and without S9 mix induced a distinct increase in
the number of revertant colonies within the range of the historical positive control data or
above.
• Fresh bacterial culture containing approximately 10E9 cells per mL were used. For approval
the titer of viable bacteria was ≥ 10E8 colonies per mL.

Assessment criteria
The test substance is considered positive in this assay if the following criteria are met:
• A dose-related and reproducible increase in the number of revertant colonies, i.e. about
doubling of the spontaneous mutation rate in at least one tester strain either without
S9 mix or after adding a metabolizing system.
A test substance is generally considered non-mutagenic in this test if:
• The number of revertants for all tester strains were within the historical negative control
range under all experimental conditions in at least two experiments carried out independently of each other.

Statistics:

not performed

Results and discussion

Test results

Species / strain:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E.coli WP2 uvrA

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

Additional information on results:

TOXICITY
A bacteriotoxic effect (decrease in the number of his+ revertants) was observed in the standard plate test depending on the strain and test conditions from about 2 500 μg/plate onwards. In the preincubation assay bacteriotoxicity (decrease in the number of his+ revertants) was observed depending on the strain and test conditions at 5 000 μg/plate.

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Test substance precipitation was found from about 100 μg/plate onwards with and without S9 mix

COMPARISON WITH HISTORICAL CONTROL DATA
The number of revertant colonies in the negative controls was within the range of the historical negative control data for each test strain with and without S9 mix. The positive control substances both with and without S9 mix induced a significant increase in the number of revertant colonies within the range of the historical positive control data or above.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative