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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information
The substance C.I. Acid blue 74 is considered to be not mutagenic in Salmonella typhimurium strain TA1535, TA1537, TA1538, TA98 and TA100 with and without metabolic activation.
Link to relevant study records
Reference
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)
Reference:
Composition 0
Qualifier:
according to
Guideline:
other: as per mentioned below
Principles of method if other than guideline:
Evaluate mutagenicity of C.I. Acid blue 74 by the standard plate-incorporation assay (Ames et al. 1975).
GLP compliance:
no
Type of assay:
bacterial gene mutation assay
Test material information:
Composition 1
Target gene:
Standard Plate-incorporation
Species / strain:
other: TA1535, TA1537, TA1538, TA98 and TA100
Details on mammalian cell lines (if applicable):
Not applicable
Additional strain characteristics:
not specified
Metabolic activation:
with and without
Test concentrations with justification for top dose:
3333, 1000, 6666 and 10000 µg/plate
Vehicle:
Vehicle- Vehicle(s)/solvent(s) used: Yes- Justification for choice of solvent/vehicle: No data available
Negative controls:
not specified
Solvent controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: sodium azide for strains TA1535 & TA100:(1.0/1µg/plate), 9-aminoacridine for TA1537 (50.0/µg/plate), and 2-nitrofluorene for TA1538 &TA98 (1.0, 2.0 or 5.0 /µg/plate) 2-aminoanthracene (2.5/µg/plate for TA1535 and TA1537, 1.0/µg/plate for TA1538 and TA98
Details on test system and conditions:
Details on test system and conditionsMETHOD OF APPLICATION: in agar (plate incorporation)DURATION- Preincubation period: No data available- Exposure duration: No data available- Expression time (cells in growth medium): No data available- 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 availableSELECTION AGENT (mutation assays): No data availableSPINDLE INHIBITOR (cytogenetic assays): No data availableSTAIN (for cytogenetic assays): No data availableNUMBER OF REPLICATIONS: No data availableNUMBER OF CELLS EVALUATED: No data availableDETERMINATION OF CYTOTOXICITY- Method: No data availableOTHER EXAMINATIONS:- Determination of polyploidy: No data available- Determination of endoreplication: No data available- Other: No data available
Evaluation criteria:
Dose-related increase in the number of revertants above spontaneous solvent controls, with a 2-fold increase for strains TA1535, TA1538, TA98 and TA100, and a 3-fold increase for TA1537.
Statistics:
No data available
Species / strain:
other: TA1535, TA1537, TA1538, TA98 and TA100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not specified
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'. Remarks: Aroclor 1254-induced male Fischer 344 rats and Syrian golden hamsters
Additional information on results:
No data available

MUTAGENICITY OF MISCELLANEOUS DYE IN THE S. typhimurium ASSAY:

Revertants per plate:

µg/plate

TA1535

TA1537

(-)S9

(R)S9

(H)S9

 (-)S9

(R)S9

(H)S9

Solvent control

23±3

9±5

15±5

9±2

10±3

9±4

Positive control

814±42

156±17

171±14

269±28

294±14

157±11

333

23±2

13±4

11±3

7±1

7±4

Toxic

1000

17±3

10±4

13±5

4±1

8±4

6±3

3333

21±2

11±2

9±3

3±2

7±3

3±1

6666

19±7

12±0

14±3

5±1

5±2

6±1

10000

19±3

22±4

10±4

3±1

Toxic

4±2

 

 

µg/plate

TA1538

 

TA98

TA100

 

 (-)S9

(R)S9

(H)S9

(-)S9

(R)S9

(H)S9

(-)S9

(R)S9

(H)S9

Solvent control

20±6

19±6

22±7

20±4

33±7

30±6

161±26

160±17

183±4

Positive control

677±27

750±33

449±23

269±12

1725±66

775±60

1104±36

2818±14

1469±35

333

10±2

17±3

27±7

18±2

23±6

32±4

160±7

147±20

171±17

1000

6±5

15±5

19±6

17±1

22±6

28±3

158±10

161±7

187±2

3333

5±3

13±3

13±3

13±6

16±7

22±3

158±16

169±11

205±48

6666

7±3

9±2

18±2

13±3

20±5

15±4

193±28

182±15

246±46

10000

7±1

7±5

13±3

23±11

17±2

15±7

167±31

198±17

201±11

Conclusions:
Interpretation of results (migrated information):negative Negative (with and without)The substance C.I. Acid blue 74 is considered to be not mutagenic in Salmonella typhimurium strain TA1535, TA1537, TA1538, TA98 and TA100 with and without metabolic activation.
Executive summary:

Plate-incorporation was carried out according to the method of Prival and Mitchell, 1982; Prival et al., 1984, to observe genetic effect of C.I. Acid blue 74 in S.typhimurium strainTA1535, TA1537, TA1538, TA98 and TA100. All strain were tested at dose 3333, 1000, 6666 and 10000 µg/plate both with and without metabolic activation (Aroclor 1254-induced male Fischer). Positive control and solvent control as used. There is no dose-related increase in the number of revertants above spontaneous solvent controls, with a 2-fold increase for strains TA1535, TA1538, TA98 and TA100, and a 3-fold increase for TA1537.Hence, the substanceC.I. Acid blue 74is considered to be not mutagenic inSalmonella typhimurium strain TA1535, TA1537, TA1538, TA98 and TA100 with and without metabolic activation.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Gene toxicity in vitro:

Peer reviewed articles were viewed to determine the mutagenic nature of the test compound C.I. Acid blue 74 (CAS no 860-22-0). The studies are summarized as below:

 

Genetic toxicity test was performed by T.P. Cameron et al, 1987 for Standard Plate-incorporation onS typhimurium(Strain TA1535, TA1537, TA1538, TA98 and TA100).All strain were tested at dose 3333, 1000, 6666 and 10000 µg/plate both with and without metabolic activation (Aroclor 1254-induced male Fischer). Positive control and solvent control as used. There is no dose-related increase in the number of revertants above spontaneous solvent controls, with a 2-fold increase for strains TA1535, TA1538, TA98 and TA100, and a 3-fold increase for TA1537.Hence, the substanceC.I. Acid blue 74is considered to be not mutagenic inSalmonella typhimurium strain TA1535, TA1537, TA1538, TA98 and TA100 with and without metabolic activation.

 

Mutagenicity testing of C.I. Acid blue 74 was performed by T.P. Cameron et al, 1987 by using the FMN preincubation modification of the Salmonella assay. Salmonella typhimurium strain TA98 and TA100 was exposed to C.I. Acid blue 74 at 33, 100, 333, 1000 and 3333 µg/plate. For metabolic activation Hamster liver $9 (30% V/V), plus cofactors (FMN, NADH, G6PD, G6P) was used and Trypan blue (228 /µg/plate) used as a positive control substance. No positive mutagenic response obtained at any tested concentration. Hence,the substance is considered to be not mutagenic inSalmonella typhimurium strain TA98 and TA100 with metabolic activation.

 

C.I. Acid blue 74 tested for mutagenicity was performed by T.P. Cameron et al, 1987, in mouse lymphoma TKassay. 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.

 

Mutagenicity of FD&C Blue no. 2 was determined by the Angela E. Auletta et al. 1977 with Salmonella typhimurium strain TA1535, TA1537, TA1538, TA98 and TA100. All strain were tested at dose 1, 10, 100, 1000 and 10000 µg/plate both with and without metabolic activation (Aroclor 1254-induced male Fischer). Positive control and solvent control as used. There is no dose-related increase in the number of revertants colonies per plate either with or without metabolic activation with either of these strains. Therefore,the substanceFD&C Blue no. 2is considered to be not mutagenic inSalmonella typhimurium strain TA1535, TA1537, TA1538, TA98 and TA100 with and without metabolic activation.

 

Reverse mutation assay was of Food blue 2 (Indigo carmine) y the M. ISHIDATE et al 1984, carried out using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 by method of Ames, McCann & Yamasaki (1975). Duplicate plates were used for each of six different concentrations of the sample. All strain were tested at dose 3333, 1000, 6666 and 10000 µg/plate both with and without metabolic activation (Aroclor 1254-induced male Fischer). All trains were exposed to six different concentrations of the chemical having 5000 µg/plate (5 mg/plate) is highest concentration. No significant increases in the number of revertant colonies were detected in any S. typhimurium strains at the maximum dose.  Therefore, the substanceFood blue 2 (Indigo carmine)is considered to be not mutagenic inSalmonella typhimurium strain TA92, TA1535, TA100, TA1537, TA94 and TA98 with and without metabolic activation.

 

Chromosomal aberration tests were performed by M. ISHIDATE et al (1984) carried out on Food blue 2 using a Chinese hamster fibroblast cell line. The cells were exposed to three different concentration having highest concentration is 12000 µg/ml (12 mg/ml). Test substance induced polyploid cells at 48 hr after treatment. On the basis of this, the substance is considered to be mutagenic in Chinese hamster fibroblast cell line without metabolic activation.

 

Mutagenicity of Food blue 2 (Indigo carmine) performed by Joseph P. Brown et al (1978) was evaluate in S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 by method of by Ames et al. All strain were tested at dose 50, 250 and 1000 µg/plate both with and without metabolic activation). No significant increases in the number of revertant colonies were detected in any S. typhimurium strains with and without metabolic activation.Therefore, the substanceFood blue 2 (Indigo carmine)is considered to be not mutagenic inSalmonella typhimurium strain TA92, TA1535, TA100, TA1537, TA94 and TA98 with and without metabolic activation.

 

Mutagenicity test was conducted to observe genetic effect ofC.I. Acid blue 74 by H. E. Seifried (2006) in S.typhimurium strainTA98, TA100, TA1535, TA1537, and TA1538 by plate Incorporation methodology. All strain were tested at dose Five doses (100-10000 µg/plate) both with and without metabolic activation (Aroclor 1254-induced male Fischer). Positive control and solvent control as used. There is no dose-related increase in the number of revertants.Hence,the substance C.I. Acid blue 74is considered to be not mutagenic inSalmonella typhimurium/TA98, TA100, TA1535, TA1537, and TA1538with and without metabolic activation.

 

C.I. Acid blue 74 tested for mutagenicity in L5178Y TK+/- 3.7.Cby H. E. Seifried (2006) inmouse lymphoma cells. 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 92-2000 µg/ml concentration of test chemical and positive control for without metabolic activation: ethyl methylsulfonate at 4.7Χ10-6 M (or methyl methanesulfonate at 10-20 µg/ mL), for metabolic activation: 3-methylcholanthrene at 1.86 Χ 10-5 M (or dimethylbenz[a]- anthracene at 0.5-4 µg/mL). There is dose related 2-fold increase in mutant frequency was observed at 92 µg/ml with S9.Hence, the substanceC.I. Acid blue 74is considered to be mutagenic inL5178Y TK +/- mouse lymphoma cells with metabolic activation. And, not mutagenic without activation.

 

From the available studies, though there was presence of two positive results but based on the majority of studies reviewed, the test material C.I. Acid blue 74 is not mutagenic in vitro.


Justification for selection of genetic toxicity endpoint
Data is from peer reveiw publications.

Justification for classification or non-classification

Gene toxicity in vitro:

Based on the key study used and its relative supporting data, the test materialC.I. Acid blue 74is not mutagenic in vitro.