29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32 copper

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Version / remarks:

26 September 2014

Deviations:

yes

Remarks:

No plasma analytics. Tested doses exceeded limit dose, 1000 cells per animal scored (but acceptable, since two groups above limit dose scored)

GLP compliance:

yes

Type of assay:

mammalian erythrocyte micronucleus test

Test material

Test material information

Constituent 1

Reference substance name:

29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32 copper

EC Number:

205-685-1

EC Name:

29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32 copper

Cas Number:

147-14-8

Molecular formula:

C32H16CuN8

IUPAC Name:

copper polyphthalocyanine

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

- Analytical purity: commercial grade
- Lot/batch No.: F 53 / H 91375

Test animals

Species:

hamster, Chinese

Strain:

other: random outbred strain

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Ciba-Geigy Tierfarm, Sisseln, Switzerland
- Age at study initiation: females 6 to 10 weeks, males 4 to 9 weeks
- Weight at study initiation: females 21 to 32 g, males 22 to 33 g in tolerability test; females 20 to 27 g, males 20 to 26 g in mutagenicity test
- Diet: NAFAG No. 924
- Water: tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature: 22 - 23 °C
- Humidity: 40 - 46 %
- Housing in air conditioned rooms
- Photoperiod: 12hrs dark / 12 hrs light

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

0.5 % Carboxymethylcellulose (CMC), Hercules Comp., USA

Details on exposure:

Tolerability test:
A preliminary test was conducted to determine the highest dosage of the test material to be applied in the mutgenicity test (Doses / Concentrations:
200, 1000 and 5000 mg/kg bw). Three groups of 4 chinese hamsters were treated with 3 different single doses. The observation period corresponded to the interval between administration and sacrifice of the animals in the mutagenicity test, plus one day. The highest dose survived by all animals was used in the second part of the tolerability test.
In the second part, the animals were treated according to the scheme used in the mutagenicity test with consecutive doses. The observation period corresponded to the interval between administration and sacrifice of the animals in the mutagenicity test, plus one day. Depending on the outcome the highest dose causing no deaths was used as the highest in the mutagenicity test.

Mutagenicity test:
The test material was administered orally to groups of 6 female and 6 male animals each. Treatment consisted of daily one application on 2 consecutive days. 24 h after the second application the animals were sacrificed.

Duration of treatment / exposure:

48 h

Frequency of treatment:

two treatments on 2 consecutive days

Post exposure period:

24 h after the second application

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

1 250 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

2 500 mg/kg bw/day (actual dose received)

Doses / concentrations 3

Dose / conc.:

5 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

Tolerability test: 2 animals per sex per dose
Mutagenicity test: 6 animals per sex per dose

Control animals:

yes, concurrent vehicle

Positive control(s):

Cyclophosphamide (ENDOXAN): 128 mg/kg bw in 20 ml/kg bw 0.5 % CMC

Examinations

Details of tissue and slide preparation:

Bone marrow was harvested from the shafts of both femurs and homogenized. Small drops were transferred on the end of a slide and spread out. 3 h later, the slides were stained in undiluted May-Grünwald solution/water for 2 min and in Giemsa´s 40 % for 20 min. After being rinsed in methanol 55 % for 5-8 sec and washed with water, the slides were cleaned in xylene and mounted in Eukitt.
The slides of three female and three male animals each of the negative and positive control group and of the groups treated with various doses of the test material were examined. 1000 bone marrow cells each were scored per animal and the following anomalies were registered: single jolly bodies, fragments of nuclei in erythrocytes, micronuclei in leucopoietic cells and polyploid cells.

Statistics:

The significance of difference was assessed by x2-test.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

In all dose groups the percentage of cells displaying anomalies of nuclei did not differ significantly from the negative control (0.1 %).
By contrast, the positive control yielded in a marked increase of the percentage of cells with anomalies (9.48 %).

Any other information on results incl. tables

Table 1: Percent of cells with anomalies of nuclei

	Animal No.	Sex (m/f)	Single Jolly Bodies	Fragments of nuclei in erythrocytes	Micronuclei in erythrocytes	Micronuclei in leucopoietic cells	Polyploid cells	Total
negative control	1	f						0.0
	2	f	0.2					0.2
	3	f	0.2					0.2
	4	m	0.1					0.1
	5	m	0.1					0.1
	6	m						0.0
cyclophosphamide	1	f	11.6	2.3	2.0			15.9
	2	f	4.5	1.0	1.6	0.1		7.2
	3	f	9.1	2.2	1.3	0.1		12.7
	4	m	6.7	0.7	1.0	0.3	0.3	9.0
	5	m	4.0	1.2	0.8			6.0
	6	m	4.4	0.6	0.9	0.2		6.1
1250 mg/kg bw	1	f						0.0
	2	f						0.0
	3	f	0.1					0.1
	4	m						0.0
	5	m	0.1					0.1
	6	m	0.1					0.1
2500 mg/kg bw	1	f						0.0
	2	f	0.2					0.2
	3	f	0.1					0.1
	4	m	0.3					0.3
	5	m	0.1					0.1
	6	m						0.0
5000 mg/kg bw	1	f						0.0
	2	f	0.2					0.2
	3	f						0.0
	4	m	0.1					0.1
	5	m						0.0
	6	m	0.1					0.1

Applicant's summary and conclusion

Conclusions:

Under the conditions of the experiment, no evidence of mutagenic effects was obtained in Chinese hamsters treated with the test material.