Perylene-3,4:9,10-tetracarboxylic dianhydride

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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

no E.coli WP2 or TA 102 was tested

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1988

Report date:

1988

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Perylene-3,4:9,10-tetracarboxylic dianhydride

EC Number:

204-905-3

EC Name:

Perylene-3,4:9,10-tetracarboxylic dianhydride

Cas Number:

128-69-8

Molecular formula:

C24H8O6

IUPAC Name:

7,18-dioxaheptacyclo[14.6.2.2²,⁵.0³,¹².0⁴,⁹.0¹³,²³.0²⁰,²⁴]hexacosa-1(23),2,4,9,11,13,15,20(24),21,25-decaene-6,8,17,19-tetrone

Details on test material:

Test materials used in this dossier are all considered to fall under the definition of nano-materials according to the European Commission Recommendation 2011/696/EU as the synthesis and manufacturing of this pigment always yields particulate material with a fine particle size distribution.

Specific details on test material used for the study:

- Name of test material (as cited in study report): persäure rein (english: peroxy acid, pure)
- Chemical nomenclature: Perylene-3,4,9,10-tetracarbonacid dianhydride
- Physical state: red powder
- Batch No.: EMHE 995

Method

Target gene:

his, trp

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

metabolising system (induced with Aroclor 1254) derived from rat liver homogenate

Test concentrations with justification for top dose:

4, 20, 100, 500, 2500, 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: see "details on test system"

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION: incubation for 48 to 72 hour at 37°C in the dark

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: bacterial growth

The first experiment was performed with all tester strains using three plates per dose to get information on mutagenicity and toxicity for calculation of an appropriate dose range. A reduced rate of spontaneously occurring colonies as well as visible thinning of the bacterial lawn were used as indicator for toxicity. Thinning of the bacterial lawn was controlled microscopically.

CONTROLS:
- without metabolic activation:
Na-azide: TA 100, TA 1535 (1µg/plate)
9-Aminoacridine: TA 1537 (50 µg/plate)
2-Nitrofluorene: TA 98, TA 1538 (2.5 µg/plate)
N-Methyl -N-nitro-N-nitrosoguanidine (MNNG): WP2uvrA (2.5 µg/plate)

- with metabolic activation:
Benzo[a]pyrene: TA 100, TA 98, TA 1538 (0.5 µg/plate), TA 1535, TA 1537 (1 µg/plate), WP2uvrA (10 µg/plate)
* 2-Aminoanthracene: TA 98, TA 100, TA 1535, TA 1537, TA 1538, WP2uvrA (10 µg/plate)

Evaluation criteria:

Not specified

Statistics:

Not specified

Results and discussion

Test results

Species / strain:

other: Salmonella typhimurium TA98, TA100, TA1535, TA1537, TA1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Toxicity and dose range finding experiment:
The test compound was tested at doses of 4 to 10000 µg/plate and proved to be not toxic to the bacterial strains at doses of 5000 µg/plate. Visible precipitation of the test compound on the plates has been observed at 500 µg/plate. Therefore, for mutagenicity testing 5000 µg/plate was chosen as the highest dose.

Any other information on results incl. tables

Mutagenicity experiment with and without metabolic activation

Dose (µg/plate)	TA 100		TA 1535		TA 1537		TA 1538		TA 98		WP2uvrA
	- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9
0	196	194	17	15	10	6	22	22	31	36	52	62
4	182	187	14	12	8	8	20	25	23	32	55	62
20	190	195	15	12	9	8	20	18	27	33	56	55
100	202	175	15	15	7	6	19	18	27	34	47	58
500 p	214	205	12	13	8	6	16	21	26	30	49	58
2500 p	204	212	17	16	6	7	13	22	24	31	50	66
5000 p	186	200	15	13	6	5	14	15	20	27	51	62
Positive Control 1	577	1108	377	122	120	151	490	596	375	905	183	344
Positive Control 2		1122		26		80		166		583		63

p = visible precipitation of the test compound on the plates

CONTROLS:

- positive control 1 without S9:

Sodium azide (1 µg/plate) for TA 100 and TA 1535

9 -Aminoacridine (50 µg/plate) for TA 1537

2 -Nitrofluorene (2.5 µg/plate) for TA 1538 and TA 98

MNNG (2.5 µg/plate) for WP2uvrA

- positive control 1 with S9:

2 -Aminoanthracene (0.5 µg/plate for TA 100, TA 1538 and TA 98; 1 µg/plate for TA 1535 and TA 1537; 10 µg/plate for WP2uvrA)

- positive control 2 with S9:

Benzo[a]pyrene (10 µg/plate) for all strains

Applicant's summary and conclusion

Conclusions:

Under the conditions chosen, the test substance was found to be not mutagenic in the reverse bacteria mutation assay.

Executive summary:

The test article was tested for its mutagenic potential in an Ames test performed under GLP and according to a protocol equivalent to OECD guidelines 471 and 472 in Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538 and TA98 and Escherichia coli WP2 uvrA in the absence and presence of metabolic activation. In the toxicity test all strains were exposed to 4 to 10000 µg/plate and proved to be not toxic to the bacterial strains at doses up to 5000 µg/plate. A standard plate test was performed in which all test strains were exposed to 4 - 5000 µg/plate. Visible precipitation of the test compound on the plates has been observed at 500 µg/plate. The test compound did not cause a significant increase in the number of revertant colonies with any of the tester strains either in the absence or presence of metabolic activation. No dose dependent effect was obtained. Under the conditions tested, the test article is not mutagenic.