Strontium 4-[(4-chloro-5-methyl-2-sulphonatophenyl)azo]-3-hydroxy-2-naphthoate (1:1)

Genetic toxicity: in vitro

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

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

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

235-471-3

EC Name:

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

Cas Number:

12238-31-2

Molecular formula:

C18H11N2O6SClMn

IUPAC Name:

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

Test material form:

solid: nanoform

Details on test material:

- 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.)

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): 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.)

Method

Target gene:

his, trp

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:

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.

Controls

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

Results and discussion

Test results

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

Any other information on results incl. tables

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