Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro gene mutation study in mammalian cells
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)

Data source

Reference
Reference Type:
publication
Title:
Mutagenic activity of 27 dyes and related chemicals in the Salmonella/microsome and mouse lymphoma TK +/- assays
Author:
T.P. Cameron, T.J. Hughes, P.E. Kirby, V.A. Fung and V.C. Dunkel
Year:
1987
Bibliographic source:
Mutation Research, 189 (1987) 223-261

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: as per mentioned below
Principles of method if other than guideline:
Evaluate mutagenicity of test substance by the mouse lymphoma TK assay.
GLP compliance:
no
Type of assay:
mammalian cell gene mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Disodium 5,5'-(2-(1,3-dihydro-3-oxo-2H-indazol-2-ylidene)-1,2-dihydro-3H-indol-3-one)disulphonate
EC Number:
212-728-8
EC Name:
Disodium 5,5'-(2-(1,3-dihydro-3-oxo-2H-indazol-2-ylidene)-1,2-dihydro-3H-indol-3-one)disulphonate
Cas Number:
860-22-0
Molecular formula:
C16H10N2O8S2.2Na
IUPAC Name:
disodium 3,3'-dioxo-1,1',3,3'-tetrahydro-2,2'-biindole-5,5'-disulfonate
Constituent 2
Reference substance name:
Indigo tin disulfonate sodium
IUPAC Name:
Indigo tin disulfonate sodium
Details on test material:
- Name of test material (as cited in study report): C.I. Acid blue 74
- Molecular formula (if other than submission substance):
C16-H10-N2-O8-S2.2Na
C16-H8-N2-Na2-O8-S2
C16-H8-N2-O8-S2.2Na
- Molecular weight (if other than submission substance): 466.3572 g/mol
- Substance type: Organic
- Physical state: Solid
Purity: 85%

Method

Target gene:
mouse lymphoma assay
Species / strain
Species / strain / cell type:
mouse lymphoma L5178Y cells
Details on mammalian cell type (if applicable):
Type and identity of media: The cells were grown in Fischer's medium for leukemic cells of mice (Gibco, Grand Island, NY) supplemented with 10% horse serum (Gibco, Grand Island, NY), antibiotics (50 U
penicillin/mi and 50 /µg streptomycin/ml), and 0.02% Pluronic F-68 (BASF Wyandotte Corp., Wyandotte, MI). All serum lots were prescreened for their ability to support optimal growth.
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: No data
- Periodically "cleansed" against high spontaneous background: : No data
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Test concentrations with justification for top dose:
Without S9: 971, 971, 1229, 1229, 1486, 1486, 1743, 1743, 2000 and 2000 µg/ml
With S9: 92, 206, 332, 439, 439, 556 and 556 µg/ml
Vehicle / solvent:
Vehicle
- Vehicle(s)/solvent(s) used: Yes
- Justification for choice of solvent/vehicle: No data available
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: Without metabolic activation: Ethyl methanesulfonate at 0.5 µl/ml with metabolic activation: 3-methylcholanthrene at 5.0 or 10.0 µg/ml.
Details on test system and experimental conditions:
Details on test system and conditions
METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: No data available
- Exposure duration: 4 hr
- Expression time (cells in growth medium): 48 hr
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: No data available

NUMBER OF CELLS EVALUATED: No data available

DETERMINATION OF CYTOTOXICITY
- Method: No data available

OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available
Evaluation criteria:
Mutant frequencies were expressed as mutants per 10⁴ surviving cells. In general, a response was considered positive if there was a dose-related increase in the mutant frequency above the spontaneous control frequency, with a 2-fold increase at more than 1 dose and relative total growth greater than 10%.
Statistics:
No data available

Results and discussion

Test results
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
No data available
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'. Remarks: Aroclor-1254-induced male Fischer 344 rats

Any other information on results incl. tables

Concentration µl/ml

S9

Relative suspension growth (%)

Relative cloning efficiency (%)

Relative total growth (%)

Average no. colonies TFT/viable

Mutant frequency (per 10⁴)

0.0

-

 

 

 

 

x0.61

971

-

100

113

112

82/230

0.71

971

-

103

108

111

71/221

0.64

1229

-

45

95

43

59/194

0.61

1229

-

102

84

86

66/172

0.7

1486

-

102

91

93

62/185

0.67

1486

-

100

96

95

49/195

0.50

1743

-

101

88

88

47/179

0.53

1743

-

93

92

86

47/187

0.50

2000

-

96

86

83

44/175

0.50

2000

-

108

94

101

55/192

0.57

Positive control

-

47

29

13

306/58

10.55

Solventᵃ

-

 

 

 

 

x0.56

 

 

 

 

Concentration µl/ml

S9

Relative suspension growth (%)

Relative cloning efficiency (%)

Relative total growth (%)

Average no. colonies TFT/viable

Mutant frequency (per 10⁴)

0.0

+

 

 

 

 

X0.53

92

+

51

45

23

131/106

2.47

206

+

37

44

16

131/102

2.57

332

+

37

49

18

137/114

2.40

439

+

34

40

14

130/94

2.77

439

+

31

46

14

124/108

2.30

556

+

26

50

13

134/116

2.31

556

+

25

40

10

125/93

2.69

Positive control

+

65

56

36

191/96

3.98

Solventb

+

 

 

 

 

X0.61

 

a: Solvent control for test chemical.

b: Solvent for positive control.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative Negative (with and without)

The substance is considered to be not mutagenic in L5178Y TK +/- mouse lymphoma cells with and without metabolic activation.
Executive summary:

Test chemical tested for mutagenicity in mouse lymphoma TK⁺′⁻ assay. toxicity of test chemical was determined both with and without S9 prepared from Aroclor-1254-induced male Fischer 344 rats. L5178Y TK +/- mouse lymphoma cells was exposed to Without S9: 971, 971, 1229, 1229, 1486, 1486, 1743, 1743, 2000 and 2000 µg/ml and positive control is Ethyl methanesulfonate at 0.5 µl/ml. With S9: 92, 206, 332, 439, 439, 556 and 556 µg/ml concentration of test chemical and positive control is 3-methylcholanthrene at 5.0 or 10.0 µg/ml. There was no dose-related increase in the mutant frequency above the spontaneous control frequency, no 2-fold increase at more than 1 dose and not relative total growth greater than 10% at any exposed concentration.

Hence, the substance is considered to be not mutagenic inL5178Y TK +/- mouse lymphoma cells with and without metabolic activation.