Manganese, 4-[(4-chloro-5-methyl-2-sulfophenyl)azo]-3-hydroxy-2-naphthalenecarboxylic acid complex

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Pigment Red 52:2(Mn) is not genotoxic in vitro based on experimental data with Pigment Red 52:2(Mn) and pigments of the same category.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Justification for type of information:

Please see the category read-across justification in the category object.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Species / strain:

Chinese hamster lung (CHL/IU)

Metabolic activation:

with and without

Genotoxicity:

other: carcinogenicity negative

Cytotoxicity / choice of top concentrations:

not specified

Species / strain:

Chinese hamster lung (CHL/IU)

Metabolic activation:

with and without

Genotoxicity:

other: polyploidy positive at precipitation

Cytotoxicity / choice of top concentrations:

not specified

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Reference 2

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Justification for type of information:

Please see the category read-across justification in the category object.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Reference 3

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

- Name of test material (as cited in study report): LITHOL Fast Maroon L 4763, Test substance No.: 06/0430-1
- Physical state: Solid (powder), red
- purity and composition: see confidential details on test item
- Impurities (identity and concentrations):
- Lot/batch No.: Standardmuster 030009P040 vom 15.07.2003
- Stability under test conditions: The stability of the test substance at room temperature in the vehicle DMSO and in water each over a period of 4 hours were determined analytically.
- Storage condition of test material: Room temperature
- Composition of test material, percentage of components: CAS-Nr. 12238-31-2: 54% (Pigment Red 52 Mn) (The dose used for testing was adjusted for the low concentration.)

Target gene:

his, trp

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:

Aroclor 1254 induced rat liver S-9 mix

Test concentrations with justification for top dose:

20 µg - 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO;
To achieve a solution of the test substance in the vehicle, the test substance preparation was treated with ultrasonic waves and was shaken thoroughly.

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

other: 2-aminoanthracene (2-AA), with S9 mix ; N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), without S9 mix; 4-nitro-o-phenylendiamine (NOPD), without S9;

Details on test system and experimental conditions:

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

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth.

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.


Toxicity detected by a
− decrease in the number of revertants
− clearing or diminution of the background lawn (= reduced his- or trp- background growth)
− reduction in the titer
is recorded for all test groups both with and without S-9 mix in all experiments.

Statistics:

The data were not statistically analyzed

Species / strain:

other: TA 1535, TA 100, TA 1537, TA 98 and E. coli WP2 uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Weak bacteriotoxic effect in the standard plate test depending on the strain and test conditions from about 4630 μg/plate onward. In the preincubation assay bacteriotoxicity was observed depending on the strain and test conditions from about 926 μg/plate

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Test substance precipitation was found from about 926 μg/plate onward.

RANGE-FINDING/SCREENING STUDIES:
In general, five doses of the test substance are tested with a maximum of 5 mg/plate, and triplicate plating is used for all test groups at least in the 1st experiment. Dose selection and evaluation as well as the number of plates used in repeat studies or further experiments are based on the findings of the 1st experiment.

COMPARISON WITH HISTORICAL CONTROL DATA:
The number of revertant colonies in the negative controls was within the normal range of the historical control data for each tester strain.
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

Standard plate test (20 - 5000 µg/plate) 1.Exp
Strain	Metabolic activation system	mean revertants in Controls	maximum revertant factor	dose dependency	Assessment
TA 98	no	29	0.9	no	negative
	yes	38	0.8	no	negative
TA 1537	no	10	0.7	no	negative
	yes	10	0.9	no	negative
TA 1535	no	16	1.0	no	negative
	yes	18	0.8	no	negative
TA 100	no	103	1.0	no	negative
	yes	118	1.0	no	negative
E.coli WP2 uvrA	no	31	1.1	no	negative
	yes	34	0.9	no	negative

Preincubation test (20 - 5000 µg/plate) 2. Exp.
Strain	Metabolic activation system	mean revertants in Controls	maximum revertant factor	dose dependency	Assessment
TA 98	no	32	0.9	no	negative
	yes	31	1.0	no	negative
TA 1537	no	9	0.9	no	negative
	yes	10	1.0	no	negative
TA 1535	no	17	1.0	no	negative
	yes	17	0.8	no	negative
TA 100	no	110	1.0	no	negative
	yes	113	1.1	no	negative
E.coli WP2 uvrA	no	32	1.3	no	negative
	yes	40	1.0	no	negative

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Only an Ames test is available for Pigment Red 52:2(Mn), however sufficient data on genotoxicity is available for read-across substances and the sulfonated aromatic amine metabolite.

 

In vitro mutagenicity in bacteria

Experimental data is available for mutagenicity in bacteria. A formulation of Pigment Red 52:2 was tested for mutagenicity in bacteria in the Ames test (GLP, OECD 471) (BASF AG 2006). The concentration was adjusted for the lower content of the pigment. The substance was tested for mutagenicity in the Salmonella typhimurium / Escherichia coli reverse mutation assay both in the standard plate test and in the preincubation test with and without the addition of a metabolizing system (S-9 mix) obtained from rat liver using the Salmonella strains TA 1535, TA 100, TA 1537, TA 98 and Escherichia coli WP2 uvrA.

 

In addition, experimental data is available data from Pigment Red 57:1(Ca), 57(Sr), 48:1(Ba), 48:2(Ca), and 48:4(Mn). These Pigments are non-mutagenic in the Ames tests.The difference lies in the sulfonated phenyl part (CAS 88-53-9) that would be derived from azo reduction of Pigment Red 52:2(Ca). This part was found to be non-mutagenic in two tests. One test was sponsored by MHWL and its short summary as well as the report in Japanese language have been published by the Japanese Competent Authority. From the available information, this study appears to be valid without restriction. The other test was sponsored by BASF in 1981 and is considered to be valid with restrictions as it was performed prior to the introduction of GLP and did not use the standard five tester strains.

 

In vitro mutagenicity in mammalian cells

Mutagenicity in mammalian cells in vitro was investigated of Pigment Red 48:2(Ca) and Pigment Red 57:1(Ca) in HPRT tests following OECD testing guideline 476 (adopted July 21, 1997) and the principles of GLP. For Pigment Red 48:2(Ca), the test was performed with a sample synthesized without additives and a purity of 99.4%. No indication of a mutagenic effect was observed at concentrations up to 720mg/plate, at least the two highest concentrations being in the precipitating range. For Pigment Red 57:1(Ca) no indication of a mutagenic effect was observed at concentrations up to 1000 µg/mL, the two highest concentrations being in the precipitating range. The absence of genotoxicity is consistent with the unscheduled DNA synthesis tests performed with both substances.

 

In vitro clastogenicity in mammalian cells

For clastogenicity in vitro, experimental data from studies with samples of Pigment Red 57:1(Ca) is taken into account. Clastogenicity in mammalian cells was investigated in two guideline (OECD 473. adopted July 21, 1997) and GLP compliant studies (Hoffmann 2008 and MHLW 1993). The highest evaluable concentrations were chosen based on visual observation of precipitates. No indication of clastogenicity or polyploidy were observed in either study. For both studies, commercial samples of known composition were tested.

Justification for classification or non-classification

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