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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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

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
Study period:
From 03 December 2012 to 21 December 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The reliability is rated 1 because the study followed the standard guideline of reference (OECD 471), which describes a procedure designed to evaluate this endpoint, the results were reviewed for reliability and assessed as valid, and the study was conducted under GLP condition.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
[bis(phenylamino)methylidene]azanium 3-[(1E)-2-[4-(phenylamino)phenyl]diazen-1-yl]benzene-1-sulfonate
EC Number:
941-151-0
Cas Number:
1690331-63-5
Molecular formula:
C31H28N6SO3
IUPAC Name:
[bis(phenylamino)methylidene]azanium 3-[(1E)-2-[4-(phenylamino)phenyl]diazen-1-yl]benzene-1-sulfonate
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): Sepisol Fast Yellow MG-DPG
- Substance type: monoconstituent
- Physical state : powder
- Stability under test conditions: stable
- Storage condition of test material: room temperature (23 +/- 5°C) and protected from light

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A pKM 101
Metabolic activation:
with and without
Metabolic activation system:
S9 from liver of rodent treated with Aroclor
Test concentrations with justification for top dose:
C1 : 0.007 mg/plate
C2 : 0.021 mg/plate
C3 : 0.062 mg/plate
C4 : 0.187 mg/plate
C5 : 0.560 mg/plate

Concentration C4 to C1 were prepared by 1:3 serial dilutions in the selected solvent from the C5 concentrations.
Vehicle / solvent:
- Solvent used: ethanol (96%)
- Justification for choice of solvent: The solubility of the test item was evaluated in a standard solvent panel (milliQ water, ethanol 96%, DMSO and corn oil). Observation of precipitation by the naked eye indicated that the test item is not soluble. The ethanol (96%) was chosen as the test item was soluble in it at a concentration of 50.0 mg/mL
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
2-nitrofluorene
sodium azide
other: 2-amino-anthracene
Remarks:
The positive controls used are different for tests with and without metabolic activation or for different tester strains (see the "section Any other information on materials and methods incl.tables" for more details)
Details on test system and experimental conditions:
METHOD OF APPLICATION: under the direct incorporation ofagar medium plate and the pre-incubation procedures

DURATION
- Preincubation period: 20 min at 37°C when the pre-incubation procedure is used.
- Exposure duration: 48 hr at 37°C
- Expression time (cells in growth medium): counting completed at the end of the exposure period.

NUMBER OF REPLICATIONS: Triplicate, in parallel with vehicale and reference item controls.

DETERMINATION OF CYTOTOXICITY
- Method: decrease in the number of revertant colonies or a clearing or diminution of the background lawn.
Evaluation criteria:
A positive result involved taking into account a dose-response effect in the range tested and/or a reproducible increase at one or more concentrations in the number of revertant colonies per plate in at least one strain with or without metabolic activation system.
The result is considered positive whenever the number of revertants of the test item treated plates is higher than those observed in the solvent treated plates, according to the following criteria:
- TA98 strain : 2 fold higher
- TA100 strain: 2 fold higher
- TA1535 strain : 3 fold higher
- TA1537 strain : 3 fold higher
- WP2 (pkM101) : 2 fold higher

The biological relevance of the results was also considered.
Statistics:
no statistic was used

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at the highest dose of 0.560 mg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A pKM 101
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at the highest dose of 0.560 mg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
COMPARISON WITH HISTORICAL CONTROL DATA:
The mean solvent control and reference item control counts complied with the laboratory historical data for each strain (see attachment)

ADDITIONAL INFORMATION ON CYTOTOXICITY:
During the cytotoxicity assay, a decrease in the number of revertant colonies > 50% compared to the solvent reference was observed indicating cytotoxicity of the test item. The lowest cytotoxic concentration was 0.560 mg/plate and used as the highest exposure concentration to perform the test (see the table in the field below)
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Results of test item cytotoxicity assay

Amount/plate

Revertants/plate

Mean

S.D.

R

Solvent:

ethanol

(96%)

-

92

80

95

89

7.9

-

Test item

Sepisol

5.00

1

2

1

1

0.6

0.0

(mg)

Fast

1.67

0

1

0

0

0.6

0.0

 

Yellow

0.56

6

4

9

6

2.5

0.1

 

MG-DPG

0.19

95

82

84

87

7.0

1.0

 

 

0.06

89

98

96

94

4.7

1.1

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

The test item does not induce point mutations or frame-shifts in the genome of the bacterial strains with or without metabolic activation regardless of the procedure. Therefore, at the an exposure dose range of 0.56 mg/plate to 0.007 mg/plate, the test item is considered to be non mutagenic/non pro-mutagenic under the experimental conditions assayed.
Executive summary:

The bacterial reverse mutation test (Ames test) assesses the mutagenic or promutagenic potential of the test item SEPISOL FAST YELLOW MG-DPG in several bacterial strains.

The test was performed in accordance with OECD Guideline 471 for the Testing of Chemicals (Bacterial Reverse Mutation Test. Adopted 21st July 1997) and the test Method B13/B14 of Commission Regulation (EC) No. 440/2008, dated May 30, 2008.

In a cytotoxicity assay, a decrease in the number of revertant colonies >50% compared to solvent reference was observed, indicating cytotoxicity of the test item. The lowest cytotoxic concentration was 0.560mg/plate and used as the highest exposure concentration for to perform the test.

Five test item doses ranging from 0.560 and 0.007 mg/plate were assayed.

None of the concentrations assayed for the test item showed an increase in the R value either with or without S9 metabolic activation regardless of the procedure.

No dose response for the test item SEPISOL FAST YELLOW MG-DPG was observed in any of the tested bacterial strains.

Based on the results obtained in this study, it can be concluded that the test item does not induce point mutations or frame-shifts in the genome of the bacterial strains with or without metabolic activation regardless of the procedure. Therefore, the test item SEPISOL FAST YELLOW MG-DPG is considered to be NON MUTAGENIC / NON PRO-MUTAGENIC under the experimental conditions assayed.