Merbromin

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

data from handbook or collection of data

Data source

Reference

Reference Type:

publication

Title:

Mutagenicity Testing of Some Drug and Cosmetic Dye Lakes with the Salmonella-Mammalian Microsome Assay

Author:

J.P. Brown, P.S. Dietrich and C.M. Bakner

Year:

1979

Bibliographic source:

Mutation Research, 66 ,181-185, 1979

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

The Ames Salmonella/mammalian-microsome assay was used to evaluate the bacterial mutagenicity of test substance.

GLP compliance:

not specified

Type of assay:

other: Salmonella/mammalian microsome assay

Test material

Test material information

Constituent 1

Reference substance name:

3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid

EC Number:

242-355-6

EC Name:

3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid

Cas Number:

18472-87-2

Molecular formula:

C20H4Br4Cl4O5.2Na

IUPAC Name:

2',4',5',7'-tetrabromo-4,5,6,7-tetrachloro-3',6'-dihydroxy-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one

Test material form:

solid: particulate/powder

Details on test material:

SOURCE OF TEST MATERIAL
- Test Item: 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No.18472-87-2)
- Source of test material: Sustainability Support Services (Europe) AB
- Lot/batch No.of test material: FG/15-16/0790
- Manufacturing Date: June, 2015
- Expiration date of the lot/batch: May, 2023
- Purity test date: No data
- Consistency: Solid, powder
- Colour: Red

RADIOLABELLING INFORMATION (Not applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Test Item was stored at ambient temperature.
- Stability under test conditions: No data
- Solubility and stability of the test substance in the solvent/vehicle: No data
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: No data

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test item was dissolved in distilled water. Test item was dissolved in distilled water.
The formulation was prepared fresh on the day of dosing.
- Preliminary purification step (if any): No data
- Final dilution of a dissolved solid, stock liquid or gel: No data
- Final preparation of a solid: No data

FORM AS APPLIED IN THE TEST (if different from that of starting material) : No data

OTHER SPECIFICS:
Safety Precautions: Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).

Specific details on test material used for the study:

- Name of test material (as cited in study report): Phloxine B
- Molecular formula: C20H2Br4Cl4O5Na2
- Molecular weight: 829.66 g/mol
- Substance type: Organic
- Physical state: Red powder
- Purity: 7-9%

Method

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA1535, TA100, TA1537, TA1538,TA98

Details on mammalian cell type (if applicable):

Not applicable.

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Liver-microsome preparation from rats injected with Aroclor 1254, 0.25 ml "S-9 mix"-plate with 0.2 ml S9/ml 'S9 mix'.

Test concentrations with justification for top dose:

10,50 and 100 µg/plate

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

ethylmethanesulphonate

methylmethanesulfonate

other: Anthragallol (For strains TA-1538 and TA98 without S9); 2-Anthramine (For all strains with S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar

DURATION
- Preincubation period: No data available
- Exposure duration:3 days

Evaluation criteria:

Mutations in Salmonella typhimurium i.e. Number of His + Revertants/plate

Statistics:

Rounded mean ± standard deviation

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA1535, TA100, TA1537, TA1538 and TA98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: Slight toxicity was noted

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

COMPARISON WITH HISTORICAL CONTROL DATA: Yes, historical negative control were used.

Remarks on result:

other: no mutagenic potential

Any other information on results incl. tables

Table: Mutagenicity Testing Of D And C Red No. 27 With The Salmonella/Microsome Test

Chemical	Dose	Microsome activation	Number of His^+Revertants/plate
			TA1535	TA100	TA1537	TA1538	TA98
Negative controls
A)		-	35±22 (265)	98 ± 26 (271)	8 ± 5 (297)	13 ± 5 (297)	33 ± 20 (282)
		±	20 ± 19 (239)	90 ± 25 (252)	11 ± 4 (283)	34 ± 15 (280)	27 ± 13 (308)
B) This study		-	42 ± 14 (9)	126 ± 24 (12)	11 ± 5 (9)	13 ± 2 (9)	24 ± 8 (9)
		±	20 ± 19 (8)	104 ± 27 (11)	12 ± 4 (9)	24 ± 6 (12)	28 ± 10 (12)
Positive controls
EMS	2µL	-	440	-	-	-	-
MMS	1µL	-	-	4860	-	-	-
9 AA	100	-	-	-	103	-	-
Anthragallol	50	-	-	-	-	47	81
2- Anthramine	1	±	824	7600	488	6400	7600
D and C Red No. 27	10	-	43	113	8	12	14^h
		±	7	114	16^h	30	24
	50	-	51	30	9	18	11
		±	6	98	12	19	23
	100	-	41	109	7	11	11^h
		±	16	90	5	19	24

 

Applicant's summary and conclusion

Conclusions:

The test substance did not showed any mutagenic activity at concentrations up to 100 μg/plate in S.typhimurium strains TA1535, TA100, TA1537, TA1538 and TA98 in the presence and absence of S9 metabolic activation system and hence, it can be concluded that test substance is non genotoxic.

Executive summary:

The Ames Salmonella/mammalian-microsome assay was used to evaluate the bacterial mutagenicity of test substance . The test substance did not showed any mutagenic activity at concentrations up to 100 μg/plate in S.typhimurium strains TA1535, TA100, TA1537, TA1538 and TA98 in the presence and absence of S9 metabolic activation system and hence the test substance is non genotoxic.