Brominated chlorinated copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, wherein the number of bromines is equal to or more than 10 and less than 16, and the number of chlorines is equal to or more than 0 and less than 7

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:

2005

Report date:

2005

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

Constituent 1

Test material form:

solid: particulate/powder

Details on test material:

green powder

Specific details on test material used for the study:

Lot 029008P050
Green powder
Storage Room temperature
Year of manufacture: 2003

Method

Target gene:

his operon

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 from liver of Aroclor 1254-induced rats

Test concentrations with justification for top dose:

0; 20; 100; 500; 2500 and 5000 microgramm/plate

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

other: 2-Aminoanthracene, N-methyl-N`-nitro-N-nitrosoguanidine, 4-nitro-phenylendiamine

Details on test system and experimental conditions:

METHOD OF APPLICATION:in agar (plate incorporation) for the first experiment and preincubation for the repeat experiment

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.

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 normal range of
the historical control data for each tester strain (see Appendix 5).
" The sterility controls revealed no indication of bacterial contamination.
" The positive control articles both with and without S-9 mix induced a significant increase in
the number of revertant colonies within the range of the historical control data or above
(see Appendix 6).
" The titer of viable bacteria was > 108/mI.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

No bacteriotoxic effect (reduced his- or trp- background growth, decrease in the number of his+ or trp+ revertants, reduction in the titer) was observed.
Test substance precipitation was found from about 100 microgramm/plate onward.
The test substance did not Iead to an increase in the number of revertant colonies either without S-9 mix or after adding a metabolizing system in two experiments carried out independently of each other (standard plate test and preincubation assay).

Any other information on results incl. tables

E.coli. WP2 uvrA	without S9				with S9
Dose/Plate	Revertants	Number	SD	Factor	Revertants	Mean	SD	Factor	Titre
DMSO	28	31	3	1.0	30	34	5	1.0	26
	31				39				30
	34				34				30
20 ug	30	34	4	1.1	31	34	3	1.0
	34				36
	37				35
100 ug	30P 27P	32	7	1.0	35P 33P	32	4	0.9
	40P				27P
500 ug	28P 29P	29	1	0.9	33P 36P	35	2	1.0
	30P				36P
2500 ug	29P	29	2	0.9	32P	32	3	0.9	21
	27P				35P				33
	30P				30P				25
5000 ug	28P	26	5	0.8	36P	32	3	0.9	27
	29P				31P				30
	20P				30P				25
4-NQO 5.0 ug	705	673	28	21.7
	661
	653
2-AA 60 ug					250	226	22	6
					207
					222

Tables from the plate incorporation test are provided in the attachment.

Applicant's summary and conclusion

Conclusions:

The substance is not mutagenic in the Ames test.