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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:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
1982-1983
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1983
Report Date:
1982
Reference Type:
publication
Title:
Unnamed
Year:
1988

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
No specific method or guideline was noted; similar to OECD guideline 471; limited documentation
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Heptane
- Analytical purity: 100% pure commercial product

Method

Target gene:
His-operon (Salmonellla), Trp-operon (E. coli)
Species / strainopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
S9 mix of livers from Aroclor1254-pretreated rats
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Metabolic activation system:
S9 mix of livers from Aroclor1254-pretreated rats
Species / strain:
E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
S9 mix of livers from Aroclor1254-pretreated rats
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
S9 mix of livers from Aroclor1254-pretreated rats
Test concentrations with justification for top dose:
max. conc. tested: 250 µg/mL
Vehicle:
- Vehicle(s)/solvent(s) used: Tween80/ethanol
Controls
Negative controls:
not specified
Solvent controls:
not specified
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: benzo[a]pyrene in DMSO, 4-nitroquinoline-N-oxide in DMSO, sodium azide, neutral red, potassium dichromate
Details on test system and conditions:
METHOD OF APPLICATION: preincubation


DURATION
- Preincubation period: no data
- Exposure duration: not applicable, preincubation method
- Expression time (cells in growth medium): 48-72 hours


DETERMINATION OF CYTOTOXICITY
- Method: other: toxicity screening test

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:
not specified
Vehicle controls valid:
not specified
Negative controls valid:
not specified
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
not specified
Negative controls valid:
not specified
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Species / strain:
E. coli WP2
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
not specified
Negative controls valid:
not specified
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
not specified
Negative controls valid:
not specified
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

The addition of heptane at amounts up to 250 µg per mL to cultures of Escherichia coli WP2 and WP2 uvr A, Salmonella typhimurium TA 1535, TA 1537, TA 1538, TA 98, and TA 100 did not lead to an increase in the reverse gene mutation frequency in any of these strains, either in the presence or in the absence of rat liver S9 fraction.

Table: Relative Reverse Mutation Rate – E. coli

Concentration

(µg/ml)

E. coliWP2

Assay 1

E. coliWP2

Assay 2

E. coliWP2

Assay 3

E. coliWP2 uvr A

Assay 1

E. coliWP2 uvr A

Assay 2

Without S9

3.91

-

1.6

0.9

-

0.9

7.81

-

2.3

1.0

-

0.9

15.6

1.2

1.3

0.8

0.5

0.8

31.3

1.1

1.4

0.7

0.8

0.6

62.5

1.0

1.0

0.8

0.9

0.5

125

0.7

0.6

1.2

0.3

0.5

250

0.5

0.8

1.0

0.2

0.5

4-nitroquinoline-N-oxide

31.7

7.7

4.8

1.9

6.3

With S9

3.91

-

1.5

0.9

-

1.0

7.81

-

2.4

0.9

-

1.4

15.6

0.7

3.1

1.0

1.8

0.9

31.3

0.7

2.8

0.8

0.8

0.9

62.5

1.0

2.4

0.6

1.0

0.9

125

0.9

2.2

1.1

2.0

0.9

250

0.7

1.3

0.9

-

0.8

4-nitroquinoline-N-oxide

1.0

3.9

0.7

9.0

19.7

Table: Relative Mutation Rate – S. typhimurium TA 1535, TA 1537, TA 1538

Concentration

(µg/ml)

TA 1535

Assay 1

TA 1535

Assay 2

TA 1537

Assay 1

TA 1537

Assay 2

TA 1538

Assay 1

TA 1538

Assay 2

TA 1538

Assay 3

Without S9

3.91

-

-

-

-

-

-

-

7.81

-

1.2

-

0.7

-

0.8

1.0

15.6

1.6

1.2

1.3

0

0.5

1.0

1.0

31.3

0.1

0.6

0.4

0

0

0.4

0.7

62.5

0

0

0

0

0

0

0

125

0

0

0

0

0

0

0

250

0

0

0

0

0

0

0

Sodium azide 1.7 µg

48.0

73.1

-

-

-

-

-

Benzo(a)-pyrene 6.7 µg

-

-

-

-

1.4

1.1

0.9

Neutral red 6.7 µg

-

-

1.5

1.6

-

-

-

With S9

3.91

-

-

-

-

-

-

-

7.81

-

0.9

-

0.7

-

-

0.8

15.6

2.1

1.0

2.4

0.7

1.5

-

1.0

31.3

1.4

0.9

1.3

0.9

2.1

-

0.7

62.5

1.7

0.6

1.8

0.9

1.8

-

1.0

125

1.7

1.3

2.1

0.6

1.2

-

1.1

250

1.1

0.7

2.3

0.8

1.3

-

0.9

Sodium azide 1.7 µg

19.3

78.3

-

-

-

-

-

Benzo(a)-pyrene 6.7 µg

-

-

-

-

2.4

-

10.6

Neutral red 6.7 µg

-

-

3.0

6.0

-

-

-

Table: Relative Mutation Rate – S. typhimurium TA 98, TA 100

Concentration

(µg/ml)

TA 98

Assay 1

TA 98

Assay 2

TA 100

Assay 1

TA 100

Assay 2

Without S9

3.91

-

0.5

-

1.1

7.81

-

0.1

-

1.0

15.6

0.6

0

1.1

0.8

31.3

0

0

1.1

0.1

62.5

0

0

1.0

0

125

0

0

1.2

0

250

0

0

0.6

0

Benzo(a)-pyrene 6.7 µg

0.8

1.2

1.6

1.0

With S9

3.91

-

0.8

-

1.0

7.81

-

1.1

-

0.9

15.6

0.9

1.1

0.9

1.0

31.3

0.9

1.0

0.8

1.1

62.5

0.8

0.9

0.7

0.9

125

0.7

0.8

0.9

1.2

250

0.4

0.5

0.9

0.9

Benzo(a)-pyrene 6.7 µg

12.6

4.7

2.8

5.4

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

The purpose of this study was to determine the mutagenicity of the test substance Normal-Heptane. A reverse mutation assay was done using S. typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100, and E. coli strains WP2 and WP2 uvr A. The strains were exposed to concentrations of 0, 3.91, 7.81, 15.6, 31.3, 62.5, 125, and 250 ug/mL for 48 -72 hrs both with and without metabolic activation. The number of revertant colonies was then counted.
No significant increases in the ratio of mutations over controls was seen. The test substance is not mutagenic in either the presence or absence of metabolic activation.
Executive summary:

The purpose of this study was to determine the mutagenicity of the test substance Normal-Heptane. A reverse mutation assay was done using S. typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100, and E. coli strains WP2 and WP2 uvr A. The strains were exposed to concentrations of 0, 3.91, 7.81, 15.6, 31.3, 62.5, 125, and 250 ug/mL for 48 -72 hrs both with and without metabolic activation. The number of revertant colonies was then counted.

No significant increases in the ratio of mutations over controls was seen. The test substance is not mutagenic in either the presence or absence of metabolic activation.