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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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

in vitro gene mutation study in bacteria
Type of information:
other: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
The test was conducted by means of Read Across approach. The reliability of the source study report is 1. Further information was attached at section 13

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Acid Black 071:2 - Similar Substance 02
Acid Black 071:2 - Similar Substance 02
Test material form:
solid: particulate/powder


Target gene:
Salmonella T: his
E. Coli: trp
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Details on mammalian cell type (if applicable):
For testing, deep-frozen (-700C to -800C) bacterial cultures (Salmonella typhimurium TA 1535, TA 100, TA 1537, TA 98 and E. coli WP2 uvrA) are thawed at room temperature, and 0.1 ml of this bacterial suspension is inoculated in nutrient broth solution (8 g Difco nutrient broth + 5 g NaCI/Iiter) and incubated in the shaking water bath at 370C for about 10 - 12 hours. As a rule, a germ density of 10^9 bacteria/mI is reached.
These cultures grown overnight are kept in iced water from the beginning of the experiment until the end in order to prevent further growth.
The use of the strains mentioned is in accordance with the current scientific recommendations for the conduct of this assay.
The Salmonella strains were obtained from KNOLL Aktiengesellschaft on October 30,
1989. The Escherichia coli strain was obtained from Merck on September 9,1991.

The Salmonella strains are checked for the following characteristics at regular intervals
deep rough character (rfa); UV sensitivity (ta, uvrB); ampicillin resistance (R factor plasmid).
E. coli WP2 uvrA is checked for UV sensitivity.
Histidine and tryptophan auxotrophy is automatically checked in each experiment via the spontaneous rate.
Metabolic activation:
with and without
Test concentrations with justification for top dose:
first experiment: 0,20,100,500,2500 and 5000 ug/plate for Ames test
second experiment: 0,4,20,100,500 and 2500 ug/plate for Prival preincubation test
Vehicle / solvent:
Untreated negative controls:
Negative solvent / vehicle controls:
Positive controls:
Positive control substance:
congo red
other: 2-aminoanthracene, benzidine, 4-nitro-o-phenylendiamine
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

- Preincubation period: 30 minutes at 30°C
- Exposure duration: 48-72h

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
Untreated negative controls validity:
Positive controls validity:
Additional information on results:
A slight decrease in the number of revertants was occasionally observed depending on the strain and test conditions from about 500 ug - 2,500 ug/plate onward. Test substance precipitatian was found from about 500 pg/plate onward.

According to the results of the present study, the test substance did not lead to an increase in the number of revertant colonies either without S-9 mix or after adding a metabolizing system in two experiments carried out independently of each other (Ames standard plate test and Prival preincubation assay).
Remarks on result:
other: all strains/cell types tested
Migrated from field 'Test system'.

Applicant's summary and conclusion

Under the experimental conditions the substance did not show any potential to be a mutagenic agent.
Executive summary:

The substance was tested for mutagenicity potential following OECD 471 (Ames test and Prival modification). Salmonella T. strains ( TA1535, TA 100, TA 1537, TA 98) and E. Coli strain (WP2 uvrA) were chosen and tested with and without S9 mix (rat and hamster livers). Negative and positive control were concurrent during the experiment. Under the experimental conditions the substance did not show any potetial to be a mutagenic agent.