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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)
Rationale for reliability incl. deficiencies:
other: Guideline study, non-GLP, strain for AT substitution was not included

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1988
Report date:
1988

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
No strain to detect AT substitution is included in the study design
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
3,3'-[ethylenebis[(3-sulpho-p-phenylene)azo]]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonic] acid, compound with 2,2',2''-nitrilotriethanol (1:6)
Cas Number:
75701-36-9
IUPAC Name:
3,3'-[ethylenebis[(3-sulpho-p-phenylene)azo]]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonic] acid, compound with 2,2',2''-nitrilotriethanol (1:6)
Test material form:
solid: particulate/powder
Details on test material:
Name of test substance (as cited in the study report): Fastusol Blau PR 8516 (gefriergetrocknet)
Storage: at 4 °C

Method

Target gene:
histidine
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
S9 fraction from Aroclor 1254 induced rat liver (prepared on Dec 7 1987)
Test concentrations with justification for top dose:
exp 1 (plate incorporation): S. typhimurium TA 1535, TA 1537, TA 98 and TA 100: 0, 20, 100, 500, 2500 and 5000 ug/plate
exp 2 (preincubation) S. typhimurium TA 1535, TA 1537, TA 98 and TA 100: 0, 20, 100, 500, 2500 and 5000 ug/plate
exp 3 (preincubation) S. typhimurium TA 98: 0, 20, 100, 500, 2500 and 5000 ug/plate
Vehicle / solvent:
aqua dist.
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
yes
Remarks:
sterility control
Positive controls:
yes
Positive control substance:
other: N-methyl-N-nitro-N-nitrosoguanidine
Remarks:
for TA 100 and TA 1535 without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
yes
Remarks:
sterility control
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylendiamine
Remarks:
for TA98 without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
yes
Remarks:
sterility control
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
for TA 1537 without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
yes
Remarks:
sterility control
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
for all strains with S9 (no additional control is included)
Details on test system and experimental conditions:
METHOD OF APPLICATION: exp 1 plate incorporation; exp 2 and 3 preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 hours at 37°C

NUMBER OF REPLICATIONS: 3 per concentration

DETERMINATION OF CYTOTOXICITY
- Method: titer/bacterial backgound lawn

S-9 fraction: ratio 3/7 as S9 fraction/cofactors prepared on the day of the experiment
concentration of cofactors:
MgCl 8 mM
KC1 33 mM
glucose-6-phosphate 5 mM
NADP 4 mM
phosphate buffer(
Evaluation criteria:
positive if:
-doubling of the spontaneous mutation rate (control)
-dose-response relationship
-reproducibility of the results
Statistics:
NA

Results and discussion

Test results
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:
slight reduction of the titer in some of the strains at 5000 ug/plate
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
The 3rd experiment was performed because in the 2nd experiment a decrease in bacterial background lawn was observed at all concentrations.

Applicant's summary and conclusion

Conclusions:
negative without metabolic activation
negative with metabolic activation

The substance is negative in an Ames test with and without metabolic activation
Executive summary:

The substance was tested at concentrations between 0 and 5000 ug/plate in an Ames test with and without metabolic activation. Both a plate incorporation and pre-incubation assay were perfomed. The substance did not induce mutations in Salmonella Typhimurium strains TA 98, TA100, TA1535 and TA1537. Therefore it is concluded that the substance is not mutagenic in bacterial cells.