Asphalt, sulfonated, sodium salt

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

N/A

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

Materials and methods were limited.

Principles of method if other than guideline:

N/A

GLP compliance:

not specified

Remarks:

HPV document indicated that the test was GLP. However, information was limited on the testing laboratory.

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Asphalt, sulfonated, sodium salt

EC Number:

269-212-0

EC Name:

Asphalt, sulfonated, sodium salt

Cas Number:

68201-32-1

Molecular formula:

UVCB

IUPAC Name:

Asphalt, sulfonated, sodium salt

Details on test material:

Chemical name: Sodium sulfonated asphalt
Purity: 100%
Lot No. 04-27-2005
- Name of test material (as cited in study report): sodium sulfonated asphalt
- Molecular formula (if other than submission substance): N/A
- Molecular weight (if other than submission substance): N/A
- Smiles notation (if other than submission substance): N/A
- InChl (if other than submission substance): N/A
- Structural formula attached as image file (if other than submission substance): N/A
- Substance type: active
- Physical state: N/A

Method

Target gene:

histidine (S. typhimurium strains) and tryptophan locus (E. coli strain)

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 100

Details on mammalian cell type (if applicable):

N/A

Additional strain / cell type characteristics:

not specified

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 98

Details on mammalian cell type (if applicable):

N/A

Additional strain / cell type characteristics:

not specified

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 1537

Details on mammalian cell type (if applicable):

N/A

Additional strain / cell type characteristics:

not specified

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 1535

Details on mammalian cell type (if applicable):

N/A

Additional strain / cell type characteristics:

not specified

Species / strain 5

Species / strain / cell type:

E. coli WP2 uvr A

Details on mammalian cell type (if applicable):

N/A

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

phenobarbital and B-naphthoflavone induced rat liver S9

Test concentrations with justification for top dose:

50, 150, 500, 1500 and 5000 ug/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: N/A

Controls

Controls 1

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO (test substance vehicle)

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracine (2AA): 1 ug/plate for TA100; 2 ug/plate for TA1535 and TA1537; 10 ug/plate for WP2uvrA-

Remarks:

With metabolic activation

Controls 2

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO (test substance vehicle)

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

With metabolic activation 5 ug/plate for TA98

Controls 3

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO (test substance vehicle)

True negative controls:

no

Positive controls:

yes

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Remarks:

Without metabolic activation 2 ug/plate for WP2uvrA-; 3 ug/plate for TA100; 5 ug/plate for TA1535

Controls 4

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO (test substance vehicle)

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

Without metabolic activation 80 ug/plate for TA1537

Controls 5

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO (test substance vehicle)

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

Without metabolic activation 0.2 ug/plate for TA98

Details on test system and experimental conditions:

METHOD OF APPLICATION: N/A


DURATION
- Preincubation period: N/A
- Exposure duration: N/A
- Expression time (cells in growth medium): N/A
- Selection time (if incubation with a selection agent): N/A
- Fixation time (start of exposure up to fixation or harvest of cells): N/A


SELECTION AGENT (mutation assays): N/A
SPINDLE INHIBITOR (cytogenetic assays): N/A
STAIN (for cytogenetic assays): N/A


NUMBER OF REPLICATIONS: 3


NUMBER OF CELLS EVALUATED: N/A


DETERMINATION OF CYTOTOXICITY
- Method: reduction in growth of bacterial background lawn


OTHER EXAMINATIONS:
- Determination of polyploidy: N/A
- Determination of endoreplication: N/A
- Other: N/A


OTHER: N/A

Evaluation criteria:

N/A

Statistics:

N/A

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: N/A
- Effects of osmolality: N/A
- Evaporation from medium: N/A
- Water solubility: N/A
- Precipitation: No test substance precipitate was observed on the plates at any of the doses tested in either the presence or absence of S9-mix. A brown colour was observed from 1500 ug/plate, but this did not prevent the scoring of revertant colonies.
- Other confounding effects: N/A


RANGE-FINDING/SCREENING STUDIES: N/A


COMPARISON WITH HISTORICAL CONTROL DATA: N/A


ADDITIONAL INFORMATION ON CYTOTOXICITY: The test substance caused no visible reduction in the growth of the bacterial background lawn at any dose level, although small decreases in revertant colony frequency were noted to several of the tester strains at 5000 ug/plate, predominantly in the presence of S9. The test substance was, therefore, tested up to the maximum recommended dose level of 5000 ug/plate.

Any other information on results incl. tables

The vehicle (dimethyl sulphoxide) control plates gave counts of revertant colonies within the normal range. All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with or without metabolic activation. Thus, the sensitivity of the assay and the efficacy of the S9 -mix were validated. No significant increases in the frequency of revertant colonies were recorded for any of the strains with any dose of the test substance, either with or without metabolic activation.

Applicant's summary and conclusion

Conclusions:

Interpretation of results:
negative No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test material, either with or without metabolic activation.

The test substance was evaluated for mutagenic potential using the Ames test. Under the conditions of the test, the test substance was determined to be non-mutagenic with or without metabolic activation.

Executive summary:

N/A