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Toxicological information

Genetic toxicity: in vitro

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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:
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

Reference
Name:
Unnamed
Type:
Constituent
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 / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535
Species / strain / cell type:
S. typhimurium TA 100
Species / strain / cell type:
S. typhimurium TA 1537
Species / strain / cell type:
S. typhimurium TA 1538
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.