[4-[4,4'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium chloride

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

1977

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Five Histidine requiring strains of Salmonella typhimurium were used in the mutagen assay. Strains TA 1535 and TA 100 are used to detect base pair substitution mutations, whereas strains TA 1537, TA 1538 and TA 98 are used to detect frameshift mutations

Principles of method if other than guideline:

Five Histidine requiring strains of Salmonella typhimurium were used in the mutagen assay. Strains TA 1535 and TA 100 are used to detect base pair substitution mutations, whereas strains TA 1537, TA 1538 and TA 98 are used to detect frameshift mutations (ames, MacCamm and Yamasaki, mutation research 31(1975) 347-364).

GLP compliance:

no

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

[4-[4,4'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium chloride

EC Number:

208-953-6

EC Name:

[4-[4,4'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium chloride

Cas Number:

548-62-9

Molecular formula:

C25H30ClN3

IUPAC Name:

4-{bis[4-(dimethylamino)phenyl]methylidene}-N,N-dimethylcyclohexa-2,5-dien-1-iminium chloride

Test material form:

solid

Details on test material:

- Name (IUPAC): 4-{bis[4-(dimethylamino)phenyl]methylidene}-N,N-dimethylcyclohexa-2,5-dien-1-iminium chloride
- Name of test material (as cited in study report): Gentian Violet
- Molecular formula: C25H30ClN3
- Molecular weight: 407.986 g/mol
- Smiles notation): CN(C)C1=CC=C(C=C1)C(=C2C=CC(=[N+](C)C)C=C2)C3=CC=C(C=C3)N(C)C.[Cl-]
- InChl : InChI=1S/C25H30N3.ClH/c1-26(2)22-13-7-19(8-14-22)25(20-9-15-23(16-10-20)27(3)4)21-11-17-24(18-12-21)28(5)6;/h7-18H,1-6H3;1H/q+1;/p-1
- Substance type: organic
- Physical state: Solid

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 100

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 1537

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 1538

Species / strain 5

Species / strain / cell type:

S. typhimurium TA 98

Metabolic activation:

with and without

Metabolic activation system:

The metabolic activation system involved the addition of 0.5ml of S-9 mixture to the chemical top agar solution.

Test concentrations with justification for top dose:

EtOH = Solvent control
11,295 = Spent Crystal Violet Isomers and Tars
2AA = 2-Aminoanthracene (positive control)

100% EtOH

11,295 µg/plate
-S.9*
2 “
4 “
8 “
10 “
2AA 5 “
10 “
10 “
100 “

Controls

Negative solvent / vehicle controls:

yes

Remarks:

EtOH

True negative controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene

Results and discussion

Test results

Species / strain:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 1538 and TA 100

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: strain/cell type: Five Histidine requiring strains

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table I

MUTAGENIC ACTIVITY OF SPENT CRYSTAL VIOLET ISOMERS ANDS TARS IN

SALMONELLA TYPHIMURIUMSTRAINS TA 1535, TA 1538, TA 98 AND TA 100 WITH METABOLIC ACTIVATION

Histidine Revertants Per Plate **
TA1535	TA1537	TA1538	TA98	TA100
17	5	17	26	125
8	7	19	15	96
14	4	17	22	125
19	4	17	25	123
18	5	51	74	153
17	5	79	81	193
17	11	94	96	185
16	11	111	129	186
				2528
361		1541	1591
	243

Table II

 

MUTAGENIC ACTIVITY OF SPENT CRYSTAL VIOLET ISOMERS ANDS TARS IN

SALMONELLA TYPHIMURIUMSTRAINS TA 1535, TA 1538, TA 98 AND TA 100 WITHOUT METABOLIC ACTIVATION

Histidine Revertants Per Plate **
TA1535	TA1537	TA1538	TA98	TA100
14	7	19	22	138
11	8	19	18	125
15	13	14	20	130
12	13	22	28	128
22	7	19	20	141
10	12	22	20	114
14	10	19	19	114
1388	11	111	129	1247
		1219	1478
	908

TABLE III

MUTAGENIC ACTIVITY OF SPENT CRYSTAL VIOLET ISOMERS AND TARS INSALMONELLA TYPHIMURIUMSTRAINS TA 1538 AND TA 98 WITH METABOLIC ACTIVATION

TRIAL II
Compound added			Histidine + Revertants Per Plate**
			TA 1538	TA98
100%E+OH 11,295 -S^.9*               2AA			17	28
	μg/plate
	5		21	12
	2		31	33
	4		61	70
	6		110	108
	8		139	124
	10		197	222
	12		221	226
	10		1998	1556
= Solvent control 11,295 = Spent Crystal Violet Isomers and Tars 2AA    = 2- Aminoanthracene (positive control) *   = Control plate without S.9 activators ** = Average number of revertants from 2 plates

Applicant's summary and conclusion

Conclusions:

Summary:
Spent Crystal violet isomers and tars were tested in Salmnella typhimurium strains TA1535, TA1537, TA1538, TA98 and TA100 in concentration up to 12g/Petri plate. The compound was mutagenic in the activated microbial assay in strains TA1538 and TA98. In strains TA 1538 this activity represents a 9-fold increase over the spontaneous mutation frequency or 16 revertants per microgram of compound tested.

Executive summary:

Spent Crystal violet isomers and tars were tested inSalmnella typhimuriumstrains TA1535, TA1537, TA1538, TA98 and TA100 in concentration up to 12mg/Petri plate. The compound was mutagenic in the activated microbial assay in strains TA1538 and TA98. In strains TA 1538 this activity represents a 9-fold increase over the spontaneous mutation frequency or 16 revertants per microgram of compound tested. All calculations are based on the average of the two trials.