[29H,31H-phthalocyaninato-N29,N30,N31,N32]cobalt

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

1

Reference substance name:

[29H,31H-phthalocyaninato-N29,N30,N31,N32]cobalt

EC Number:

222-012-7

EC Name:

[29H,31H-phthalocyaninato-N29,N30,N31,N32]cobalt

Cas Number:

3317-67-7

Molecular formula:

C32H16N8Co

IUPAC Name:

[29H,31H-phthalocyaninato-N29,N30,N31,N32]cobalt

Method

Target gene:

His D for Salmonella typhimurium TA 98
His C for Salmonella typhimurium TA 1537
His G for Salmonella typhimurium TA 100 and TA 1535
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 mix (microsome fraction prepared from Sprague Dawley rat liver homogenate)

Test concentrations with justification for top dose:

50 µg/ plate
150 µg/ plate
500 µg/ plate
1500 µg/ plate
5000 µg/ plate
A preliminary cytotoxicity testing (using strain TA 100) has been performed. In case bacteriostatic activity is detected, the highest concentration to be retained is that exhibiting a bacteriostatic activity of 75% or less. None of the concentration exhibit bacteriostatic effect.

Vehicle / solvent:

Homogeneous suspension has been prepared using ethanol.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive control substance:

7,12-dimethylbenzanthracene

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-Anthramine (with metabolic activation), cis-Platinum (II) Diammine Dichloride (without metabolic activation)

Details on test system and experimental conditions:

Two assays are performed : the first without and with metabolic activation (direct incorporation method). The test is then repeated in an independent assay. Depending on the results at the first test, the same method is used, but the test dose range may be modified and/ or in presence of metabolic activation, the indirect or direct incorporation method may be performed as explain.
Three plate per concentration are studied and are incubated at 37°C over a 48-72 hour period. The number of revertant colonies per plate is counted.
Date are presented as the number of revertant colonies (mean +/- standard deviation) per plate.

Assay repetition:
If the first assay is positive, the second one is performed in the same manner.
If the first assay, in the presence of test item, is negative, the pre-incubation is performed for the second assay.

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 98

Metabolic activation:

with and without

Genotoxicity:

positive

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 100

Metabolic activation:

with

Genotoxicity:

positive

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 100

Metabolic activation:

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:

other: The others strain (TA 1535, TA 1537, Escherichia coli WP2)

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

Remarks on result:

other: Mutagenicity activity from 150 to 5000 µg/plate without S9 and from 500 to 5000 µg/plate with S9

Applicant's summary and conclusion

Conclusions:

According to OECD 471, GLP study, mutagenic activity is observed on Salmonella thyphimurium TA 98 (without, or with metabolic activation) and in Salmonella typhimurium TA 100 (with metabolic activation), therefore, Cobalt Phthalocyanine is considered mutagenic on bacteria.