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

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2017-04-07 to 2017-07-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

1

Reference substance name:

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

EC Number:

309-627-7

EC Name:

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

Cas Number:

100545-46-8

Molecular formula:

C32H16-τN8Cu.(SO3)m.(SO3H)p.(C9H19N2SO2)O.(C9H20N2SO2)n

IUPAC Name:

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

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature

Method

Target gene:

His D for Salmonella typhimurium TA 98
His C for Salmonella typhimurium TA 1537
His G for Salmonella typhimurium TA 1535 and TA 100
Tryp E for Escherichia coli WP2 (uvr A-) (pKM101)

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

S9 fraction, microsome fraction prepared from Sprague Dawley rat liver homogenate

Test concentrations with justification for top dose:

50, 150, 500 1500 and 5000 µg/plate
Selection of the top dose: The preliminary cytotoxicity test performed on the TA 100 showed that neither original solutions nor dilutions have bacteriostatic effect.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: ethanol was found to be an appropriate solvent, to obtain an homogeneous suspension

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

other: Cis-Platinum (II), 2-anthramine and Dimethyl-anthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION:
- First assay: direct incorporation method, +/- S9 mix
- Second assay: pre-incubation test method +/- S9 mix

DURATION
- Preincubation period: 30 min at 37°C preincubation procedure used.
- Exposure duration:48-72 hr at 37°C
- Expression time (cells in growth medium): counting completed at the end of the exposure period in each plate


NUMBER OF REPLICATIONS: Triplicate, in parallel with negative, solvent and positive controls.

DETERMINATION OF CYTOTOXICITY
- Method: decrease in the number of revertant colonies or a clearing or diminution of the background lawn.

Evaluation criteria:

Ensure that the criteria validity of the study are well respected namely :
- The bacteriostatic activity of the highest concentration tested shall be equal or less than 75%
- The spontaneous reversion rate of the absolute negative control shall comply with the historical values of the laboratory
- The spontaneous reversion rate of the solvent shall not be statistically different from absolute negative control
- The mean number of revertant colonies obtained for each strain and the corresponding positive control, with and/ or without metabolic activation shall comply with the historical values of the laboratory
- Negative and positive values should not show significant difference with the historical values of the laboratory (+/- 2 standards deviation)

Statistics:

Not provided

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Doses (5000, 1500, 500, 150 and 50 µg/plate) performed from suspensions of the test item SEPISOL FAST BLUE GD BATCH: 616587 (LEMI code : GJV240317-2), provided by BIMA 83 do not induce any mutagenic change in Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100 and in Escherichia coli WP2(uvrA-)(pKM 101) without or with metabolic activation, according to the OECD Guidelines n°471.

Executive summary:

Asssay: Bacterial reverse mutation test using "Salmonella typhimurium his-" and "Escherichia coli" WP2(uvrA-)(pKM101) according to OECD n°471 (LEMI operating procedure n°MB08/45).

Suspensions (GJV020517-S2 and GJV090517-S2) obtained from GJV240317-2, have been tested for their capacity to induce reverse mutation in four Salmonella typhimurium strains and one Escherichia coli WP(uvrA-)(pKM101) strain. This study was performed in the absence and presence of metabolic activation. Two independent assays were carried out.

For assay n°1, various concentrations of GJV020517-S2 were put in contact with the strains in the absence and presence of metabolic activation system (S9 -mix 10% (v/v)).

For assay n°2, various concentrations of GJV090517-S2 were put in contact with the strains in the absence of metabolic activation and with pre-activation in the presence of metabolic activation system (S9 -mix 10% (v/v)).

For the two assays, negative and positive controls were carried out in parallel. Positive controls induced a significant increase in the number of revertant colonies compared to negative controls. There is no significant difference between the number of spontaneous reversions, the number of reversions obtained in the positive controls (without and with metabolic activation), and the mean of corresponding experimental "historical data" values obtained in the laboratory.

These results validate the two tests.

There is no evidence of any increase in the number of revertant colonies in the presence of the various concentration of the test item (5000, 1500, 500, 150 and 50 µg/plate) without and with metabolic activation in Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100 and in Escherichia coli WP2(uvrA-)(pKM 101).

Conclusion:

Doses (5000, 1500, 500, 150 and 50 µg/plate) prepared from suspensions of the test item do not induce any mutagenic change in Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100, and in Escherichia coli WP2(uvrA-)(pKM 101) without, or with metabolic activation, according to the OECD Guidelines n°471.