COAGULANT 122 (SOLID)

Genetic toxicity: in vitro

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:

08.12.1994-22.12.1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Details on test material:

Name of test material (as cited in study report): Coagulant 122 (solid)
Substance type: Clear slightly viscous yelow liquid
Physical state: 80.4% solid
Batch number: 9435262-8489-A
Storage condition of test material: 4°C in the dark under nitrogen, desiccated.

Method

Target gene:

In this in vitro assessment of the mutagenic potential of Coagulant 122 (solid), histidine dependent auxotrophic mutants of Salmonella typhimurium (strains TA 1535, TA 1537, TA 98 and TA 100) were exposed to the test substance, diluted in water which was also used as a negative control.

Species / strain

Species / strain / cell type:

bacteria, other: Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver S9

Test concentrations with justification for top dose:

Preliminary Toxicity Assay: (with metabolic activation): 5, 50, 500, 5000 µg/plate
Mutagenic Assay: (without metabolic activation): 50 , 150, 500, 1500, 5000 µg/plate

Vehicle / solvent:

Solvent: Water

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

(DMSO)

True negative controls:

no

Positive controls:

yes

Remarks:

5000 µg/plate was chosen as the top dose level in the mutation tests.

Positive control substance:

ethylmethanesulphonate

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

(DMSO)

True negative controls:

no

Positive controls:

yes

Remarks:

5000 µg/plate was chosen as the top dose level in the mutation tests.

Positive control substance:

cyclophosphamide

Results and discussion

Test results

Species / strain:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

(Toxicity was generally observed at 5000 µg/plate with the plate incorporation method and from 5 to 5000 µg/plate with Samonella with the preincubation method.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Observations:
There were no increases in the number of revertant colonies.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

The revertant colony counts for Coagulant 122 (solid) obtained in the preliminary toxicity test are shown in Table 1. Coagulant 122 (solid) was not toxic towards the tester strains. Therefore 5000 µg/plate was chosen as the top dose level in the mutation tests.
The mean number of revertant colonies, together with the individual plate counts for Coagulant 122 (solid) obtained in the first mutation test with the tester strains are shown in Table 2. positive control mutability checks are shown in Table 3.
The mean number of revertant colonies, together with the individual plate counts for Coagulant 122 (solid) obtained in the second mutation test with the tester strains ari shown in Table 4. positive control mutability checks are shown in Table 5.
No substantial increases in revertant colony numbers of any of the tester strains were observed following treatment with Coagulant 122 (solid) at any dose level, in the presence or absence of S-9 mix, in either mutation test.
The concurrent positive control compounds demonstrated the sensitivity of the assay and the metabolising activity of the liver preparations.
It is concluded that, when tested in water, Coagulant 122 (solid) shows no evidence of mutagenic activity in this bacterial system.

Executive summary:

In this in vitro assessment of the mutagenic potential of Coagulant 122 (solid), histidine dependent auxotrophic mutants of Salmonella typhimurium (strains TA 1535, TA 1537, TA 98 and TA 100) were exposed to the test substance, diluted in water which was also used as a negative control.

Two independent mutation test were performed, in the presence and absence of liver preparations from Aroclor 1254-induced rats. In the preliminary toxicity test with dose levels of up to 5000 µg/plate no toxicity was observed. A top dose level of 5000 µg/plate was chosen for the subsequent mutation study. Other dose levels used in the mutation assays were: 1500, 500, 150, 50 µg/plate.

No evidence of mutagenic activity was seen at any dose level of Coagulant 122 (solid) in either mutation test.

The concurrent positive control compounds demonstrated the sensitivity of the assay and the metabolising activity of the liver preparations. It is concluded that, when tested in water, Coagulant 122 (solid) was not mutagenic in this bacterial system.