Hydrogen [29H,31H-phthalocyaninesulphonato(3-)-N29,N30,N31,N32]cuprate(1-), compound with dodecylamine (1:1)

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The test substance was not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay (OECD 471, GLP) and the HPRT-Test (OECD 476, GLP). It is considered to be non clastogenic in a weight-of-evidence approach.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-compliant guideline study, available as unpublished report, no restrictions, fully adequate for assessment.

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Target gene:

his-locus

Species / strain / cell type:

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

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver S9 mix

Test concentrations with justification for top dose:

1st experiment: 0; 20; 100; 500; 2500 and 5000 µg/plate
2nd experiment: 0; 500; 1000; 1500; 2000 and 2500 µg/plate
3rd experiment: 0; 0.8; 4; 20; 100 and 500 µg/plate

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 selected as the vehicle, which had been demonstrated to be suitable in bacterial reverse mutation tests and for which historical control data are available.

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-methyl-N'-nitro-N-nitrosoguanidine

Remarks:

strains TA 1535 and TA 100, without S9 mix

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylendiamine

Remarks:

strain TA 98, without S9 mix

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

strain TA 1537, without S9 mix

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

strain E. coli WP2 uvrA, without S9 mix

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

all strains, with S9 mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION
- Preincubation period: 20 minutes at 37°C (preincubation method only)
- Exposure duration: 48-72 hours at 37°C

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: decrease in the number of revertants, clearing or diminution of the background lawn (= reduced his- or trp- background growth), reduction in the titer

Evaluation criteria:

The test chemical 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 S-9 mix or after adding a metabolizing system.

A test substance is generally considered nonmutagenic in this test if:
* The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments carried out independently of each other.

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

from 500 - 2500 µg/plate onward (standard plate test) and from 100 µg/plate (preincubation test) (strain and test condition dependent)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

from 500 - 2500 µg/plate onward (standard plate test) and from 100 µg/plate (preincubation test) (strain and test condition dependent)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Test substance precipitation was found from about 500 µg/plate onward.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

In a GLP-compliant study, performed according to OECD guideline 471, the test substance was tested for its mutagenic potential based an the ability to induce point mutations in selected loci of several bacterial strains, i.e. Salmonella typhimurium and Escherichia coli, in a reverse mutation assay.The assay was performed in two independent experiments both with and without liver microsomal activation. Each concentration and the controls were tested in triplicate.The test article was tested at the following concentrations: 20.0- 5000 µg/plate(SPT) and 0.8 - 500 µg/plate(PIT).

Precipitation of the test substance was found from about 500 µg/plate onward. A bacteriotoxic effect (reduced background growth, decrease in the number of revertants, reduction in the titer) was observed in the standard plate test depending on the strain and test conditions from about 500 - 2500 µg/plate onward. In the preincubation assay bacteriotoxicity was observed depending on the strain and test conditions at doses> 100 µg/plate. According to the results of the present study, the test substance did not lead to an increase in the number of revertant colonies either without S-9 mix or after adding a metabolizing system in three experiments carried out independently of each other (standard plate test and preincubation assay). Thus, under the experimental conditions chosen here, it is concluded that the substance not a mutagenic agent in a bacterial reverse mutation test.

The substance was assessed for its potential to induce gene mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus in Chinese hamster ovary (CHO) cells in vitro (OECD 476, GLP). Two independent experiments were carried out, both with and without the addition of liver S9 mix from phenobarbital- and β-naphthoflavone induced rats (exogenous metabolic activation). Based on an initial range-finding cytotoxicity test for the determination of the experimental doses and taking into account the cytotoxicity actually found in the main experiments,concentrations up to 0.5 mg/ml were tested.Following attachment of the cells for 20 - 24 hours, cells were treated with the test substancefor 4 hours in the absence and presence of metabolic activation. Subsequently, cells were cultured for 6 - 8 days and then selected in 6-thioguanine-containing medium for another week.

Finally, the colonies of each test group were fixed with methanol, stained with Giemsa and counted. The negative controls gave mutant frequencies within the range expected for the CHO cell line. Both positive control substances, ethyl methanesulfonate (EMS) and 7,12-dimethylbenz[a]anthracene (DMBA), led to the expected increase in the frequencies of forward mutations.

The test substance was poorly soluble. Hence homogeneous test substance preparations in culture medium were add to the cultures. In both experiments in the absence and the presence of metabolic activation at least the highest concentrations tested for gene mutations were clearly cytotoxic. Based on the results of the present study, the test substance did not cause any biologically

relevant increase in the mutant frequencies either without S9 mix or after the addition of a metabolizing system in two experiments performed independently of each other.

Regarding clastogenicity, experimental data on the target substance is not available. No structural alert for protein binding or DNA damage was identified by the profilers in the OECD QSAR toolbox v3.3.0.152.

(Q)SAR modelling using the software TIMES-predicted the substance to be non clastogenic, with the limitation that the structure is out of the applicability domain of the model as it contains a number of fragments unknown to the training set of the model.

No hazard for clastogenicity was identified for copperphthalocyanine both in valid in vitro and in vivo studies. No hazard for clastogenicity in vivo (OECD 474, GLP) was identified for the higher substituted analogue (CAS 108300-90-9). Although the substance has a higher molecular weight, it caused systemic toxicity at lower doses than the target substance in the subacute oral toxicity study in rats. It is therefore considered suitable for the assessment of clastogenicity in vivo. 

Absence of clastogenicity in vivo was also observed for a similar UVCB substance (CAS81457-65-0)which has a sulphonamide bond instead of an ionic bond and which contains an excess of free sulonic acid groups. It is of lower water but of much higher octanol solubility. In the in vivo micronucleus study, it was applied using peanut oil as a vehicle. In this vehicle and species, it caused mortality at high doses indicating that systemic uptake has taken place.

 

 

Reliable experimental data on related alkylamine cations is described in the US HPV programme and its published robust study summaries (Doc no. 201-14978 December 29, 2003). For example,no hazard for clastogenicity was identified for the analogue amine (Cis-9-Octadecenylamine, CAS 112-90-3).

 

 

Taking into account all available data, the target substance is considered to be non clastogenic in a weight-of-evidence approach. A datamatrix and a more detailed read-across justification is attached both in this endpoint summary and to the Chemical Safety report.

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available studies are considered reliable and suitable for classification purposes under 67/548/EEC. As a result the substance is not considered to be classified for genetic toxicity under Directive 67/548/EEC.

 

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for genetic toxicity under Regulation (EC) No. 1272/2008.