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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
Study period:
September - October 1982
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test procedure according to national standards
Justification for type of information:
The justification for read across is provided as an attachment in IUCLID Section 13.
Cross-referenceopen allclose all
Reason / purpose:
read-across: supporting information
Reference
Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
September - October 1982
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test procedure according to national standards
Justification for type of information:
The justification for read across is provided as an attachment in IUCLID Section 13.
Reason / purpose:
read-across source
Reason / purpose:
reference to same study
Key result
Species / strain:
S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98, TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
Concentrations up to 8000 µg/mL in agar layer cultures did not increase the reverse mutation frequency of S. typhimurium strains either in the presence or absence of a rat liver microsomal activation system.
Cytotoxicity / choice of top concentrations:
other: There was a differential toxicity of the test material to individual strains of Salmonella and the reduction of toxicity in the presence of an S9 mix
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
E. coli, other: WP2, WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
Concentrations up to 8000 µg/mL in agar layer cultures did not increase the reverse mutation frequency of E. coli strains either in the presence or absence of a rat liver microsomal activation system
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
There was no evidence of test compound precipitation in the assay system.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Table: Relative Mutation Rate – Bacterial Experiment I without Metabolic Activation

Concentration

E. coli WP2

E. coli WP2 uvr A

S. typhimurium TA 1535

S. typhimurium TA 1537

S. typhimurium TA 1538

S. typhimurium TA 98

S. typhimurium TA 100

Without S9

3.91

-

-

-

-

-

-

-

7.81

-

-

-

-

-

-

-

15.6

-

-

-

-

-

-

-

31.3

-

-

-

-

-

-

-

62.5

0.8

0.7

1.2

0.1

0.7

0.2

1.4

125

0.9

0.6

0.7

0

0.4

0

0.7

250

0.7

0.8

0.6

0

0

0

0.3

500

0.7

0.7

0.9

0

0

0

0.1

1000

0.8

0.5

0.9

0

0

0

0

2000

0.7

0.6

0.7

0

0

0

0

4000

0.8

0.6

0.6

0

0

0

0

8000

0.6

0.5

0.7

0

0

0

0

Sodium azide 2 µg

-

-

89.8

-

-

-

-

Benzo(a)-pyrene 7 µg

-

-

-

-

1.0

0.7

0.9

4-nitroquinoline-N-oxide 3 µg

39.8

3.2

-

-

-

-

-

Neutral red 7 µg

-

-

-

1.8

-

-

-

Table: Relative Mutation Rate – Bacterial Experiment I with Metabolic Activation

Concentration

E. coli WP2

E. coli WP2 uvr A

S. typhimurium TA 1535

S. typhimurium TA 1537

S. typhimurium TA 1538

S. typhimurium TA 98

S. typhimurium TA 100

Without S9

31.3

-

-

-

-

-

-

-

62.5

0.7

1.3

1.4

0.3

1.5

0.7

1.0

125

0.8

1.0

1.1

0.2

1.3

1.3

0.8

250

0.9

1.0

0.8

0.6

1.5

1.2

1.0

500

0.7

0.8

1.0

0.2

1.0

0.7

1.3

1000

0.7

0.7

1.0

0.4

0.7

0.2

1.4

2000

1.1

0.9

1.1

0

0.4

0

1.2

4000

1.0

1.0

1.0

0

0.7

0

1.1

8000

0.8

1.0

0.5

0

0.7

0

1.1

Sodium azide 2 µg

-

-

15.2

-

-

-

-

Benzo(a)-pyrene 7 µg

-

-

-

-

3.7

5.3

3.1

4-nitroquinoline-N-oxide 3 µg

1.2

3.6

-

-

-

-

-

Neutral red 7 µg

-

-

-

4.4

-

-

-

Table: Relative Mutation Rate – Bacterial Experiment II without Metabolic Activation

Concentration

E. coli WP2

E. coli WP2 uvr A

S. typhimurium TA 1535

S. typhimurium TA 1537

S. typhimurium TA 1538

S. typhimurium TA 98

S. typhimurium TA 100

Without S9

3.91

-

-

-

1.1

-

0.8

-

7.81

-

-

-

0.6

-

0.9

-

15.6

-

-

-

0.7

0.9

0.8

-

31.3

-

-

-

0.4

1.0

0.8

1.1

62.5

1.0

1.0

1.2

0.4

0.7

0.4

1.0

125

1.0

1.0

0.9

0

0.5

0

0.9

250

0.7

0.8

1.4

-

0.2

-

1.0

500

0.9

0.7

1.1

-

0

-

0.7

1000

1.0

0.8

0.9

-

0

-

-

2000

0.7

0.9

0.7

-

-

-

-

4000

0.9

0.8

0.9

-

-

-

-

8000

0.8

0.7

0.9

-

-

-

-

Sodium azide 2 µg

-

-

86.6

-

-

-

-

Benzo(a)-pyrene 7 µg

-

-

-

-

1.1

1.1

1.3

4-nitroquinoline-N-oxide 3 µg

36.7

5.3

-

-

-

-

-

Neutral red 7 µg

-

-

-

1.2

-

-

-

Table: Relative Mutation Rate – Bacterial Experiment II with Metabolic Activation

Concentration

E. coli WP2

E. coli WP2 uvr A

S. typhimurium TA 1535

S. typhimurium TA 1537

S. typhimurium TA 1538

S. typhimurium TA 98

S. typhimurium TA 100

Without S9

31.3

-

-

-

1.1

0.9

0.8

-

62.5

0.9

0.9

0.7

1.2

1.1

1.0

1.1

125

1.1

1.0

1.2

1.1

1.1

0.9

1.0

250

1.4

0.8

0.8

1.4

1.1

0.8

1.1

500

1.2

1.0

0.9

1.0

1.1

0.8

1.2

1000

1.0

1.1

0.8

0.5

0.9

0.4

0.9

2000

0.8

1.0

1.1

0

0.9

0.1

0.9

4000

0.8

0.9

0.8

-

0.5

-

0.9

8000

1.0

0.7

0.7

-

0.4

-

0.9

Sodium azide 2 µg

-

-

18.2

-

-

-

-

Benzo(a)-pyrene 7 µg

-

-

-

-

9.7

6.1

6.5

4-nitroquinoline-N-oxide 3 µg

1.1

2.8

-

-

-

-

-

Neutral red 7 µg

-

-

-

5.2

-

-

-

Conclusions:
Interpretation of results:
negative with metabolic activation
negative without metabolic activation

The purpose of this study was to determine the mutagenicity of the test substance hydrocarbons, C7-C9, n-alkanes, isoalkanes, cyclics. An Ames reverse mutation assay was done on S. typhimurium strains, TA 1535, TA 1537, TA 1538, TA 98 and TA 100, and E. coli strains WP2 uvr A and WP2, both with and without metabolic activation. Species were tested at concentrations ranging from 0.31 to 8000 ug/plate. The test plates were incubated for 48 -72 hrs, and then the number of colonies were counted. For all species, there was no significant increase in the number of revertants as compared to negative controls. Positive controls showed a significant increase in mutations. The test substance is not mutagenic.
Executive summary:

This data is being read across from the source study that tested Hydrocarbons, C7-C9, n-alkanes, isoalkanes, cyclics based on analogue read across.

The purpose of this study was to determine the mutagenicity of the test substance hydrocarbons, C7-C9, n-alkanes, isoalkanes, cyclics. An Ames reverse mutation assay was done on S. typhimurium strains, TA 1535, TA 1537, TA 1538, TA 98 and TA 100, and E. coli strains WP2 uvr A and WP2, both with and without metabolic activation. Species were tested at concentrations ranging from 0.31 to 8000 ug/plate. The test plates were incubated for 48 -72 hrs, and then the number of colonies were counted. For all species, there was no significant increase in the number of revertants as compared to negative controls. Positive controls showed a significant increase in mutations. The test substance is not mutagenic.

Reason / purpose:
reference to same study

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1983
Report Date:
1983
Reference Type:
publication
Title:
The genetic toxicology of some hydrocarbon and oxygenated solvents
Author:
Brooks, T.M. et al.
Year:
1988
Bibliographic source:
Mutagenesis 3(3): 227-232

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
- limited documentation of cytotoxicity data
GLP compliance:
no
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): SBP 100/140 (Special Boiling Point Spirit 100/140)
- Substance type: clear, colourless liquid
- Physical state: liquid
- Analytical purity: 100% pure commercial product
- Composition of test material, percentage of components: Special Boiling Point Spirits, SBP 100/140 is a water-white mixture of paraffins and cyclo-paraffins [65% n and isoparaffins, and 35% naphthenes] in the C7-C9 range with 0.01% aromatic hydrocarbons.
- Lot/batch No.: ex Tank 77 P; Indent 9200/9515; Ref LONO42279
- Stability: verified by IR spectra on 14 July 1982 and 20 January 1983; no differences between two spectra
- Storage condition of test material: stored in the dark at ambient temperatures
- Other: Code Number: Q.5811; IP 446/82
- Source: Shell Nederland Raffinaderij B.V., Rotterdam
- test material received on 9 July 1982

Method

Target gene:
S. typhimurium strains: his-operon
E. coli strains: tryptophan-operon
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98, TA 100
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
E. coli, other: WP2, WP2 uvr A
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
Rat liver homogenate S9 activation, 10 % S9 liver homogenate from Aroclor treated rats
Test concentrations with justification for top dose:
TA 1538 (-S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000; (+S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000 µg/mL
TA 1538 (-S) mix): 0, 15.62, 31.25, 62.5, 125, 250, 500, 1000; (+S9 mix): 0, 31.25, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000 µg/mL
TA 1537 (-S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000; (+S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000 µg/mL
TA 1537 (-S9 mix): 0, 3.9, 7.81, 15.62, 31.25, 62.5, 125; (+S9 mix): 0, 31.25, 62.5, 125, 250, 500, 1000, 2000 µg/mL
TA 1535 (-S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000; (+S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000 µg/mL
TA 100 (-S9 mix): 0, 0.31, 0.62, 1.25, 2.5, 5, 10; (+S9 mix): 0, 7.81, 15.62, 31.25, 62.5, 125, 250 µg/mL
TA 100 (-S9 mix): 0, 31.25, 62.5, 125, 250, 500; (+S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000 µg/mL
TA 100 (-S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000; (+S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000 µg/mL
TA 98 (-S9 mix): 0, 0.31, 0.62, 1.25, 2.5, 5, 10; (+S9 mix): 0, 7.81, 15.62, 31.25, 62.5, 125, 250 µg/mL
TA 98 (-S9 mix): 0, 3.9, 7.81, 15.62, 31.25, 62.5, 125; (+S9 mix): 0, 31.25, 62.5, 125, 250, 500, 1000, 2000 µg/mL
TA 98 (-S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000; (+S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000 µg/mL
EC WP2 (-S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000; (+S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000 µg/mL
EC WP2uvrA (-S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000; (+S9 mix): 0, 62.5, 125, 250, 500, 1000, 2000, 4000, 8000 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Tween 80 and ethanol
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: TA 1538, TA 98, TA 100: benzo(a)pyrene; TA 1537: neutral red; TA 1535: Sodium azide; and E. coli strains: 4-nitroquinoline-N-oxide
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 30 min
- Incubation period: 48-72 hours

NUMBER OF REPLICATIONS: All test were carried out in triplicate.

Two replicate assays were carried out on different days to confirm reproducibility of results.
Statistics:
none

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98, TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
Concentrations up to 8000 µg/mL in agar layer cultures did not increase the reverse mutation frequency of S. typhimurium strains either in the presence or absence of a rat liver microsomal activation system.
Cytotoxicity / choice of top concentrations:
other: There was a differential toxicity of the test material to individual strains of Salmonella and the reduction of toxicity in the presence of an S9 mix
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
E. coli, other: WP2, WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
Concentrations up to 8000 µg/mL in agar layer cultures did not increase the reverse mutation frequency of E. coli strains either in the presence or absence of a rat liver microsomal activation system
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
There was no evidence of test compound precipitation in the assay system.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table: Relative Mutation Rate – Bacterial Experiment I without Metabolic Activation

Concentration

E. coli WP2

E. coli WP2 uvr A

S. typhimurium TA 1535

S. typhimurium TA 1537

S. typhimurium TA 1538

S. typhimurium TA 98

S. typhimurium TA 100

Without S9

3.91

-

-

-

-

-

-

-

7.81

-

-

-

-

-

-

-

15.6

-

-

-

-

-

-

-

31.3

-

-

-

-

-

-

-

62.5

0.8

0.7

1.2

0.1

0.7

0.2

1.4

125

0.9

0.6

0.7

0

0.4

0

0.7

250

0.7

0.8

0.6

0

0

0

0.3

500

0.7

0.7

0.9

0

0

0

0.1

1000

0.8

0.5

0.9

0

0

0

0

2000

0.7

0.6

0.7

0

0

0

0

4000

0.8

0.6

0.6

0

0

0

0

8000

0.6

0.5

0.7

0

0

0

0

Sodium azide 2 µg

-

-

89.8

-

-

-

-

Benzo(a)-pyrene 7 µg

-

-

-

-

1.0

0.7

0.9

4-nitroquinoline-N-oxide 3 µg

39.8

3.2

-

-

-

-

-

Neutral red 7 µg

-

-

-

1.8

-

-

-

Table: Relative Mutation Rate – Bacterial Experiment I with Metabolic Activation

Concentration

E. coli WP2

E. coli WP2 uvr A

S. typhimurium TA 1535

S. typhimurium TA 1537

S. typhimurium TA 1538

S. typhimurium TA 98

S. typhimurium TA 100

Without S9

31.3

-

-

-

-

-

-

-

62.5

0.7

1.3

1.4

0.3

1.5

0.7

1.0

125

0.8

1.0

1.1

0.2

1.3

1.3

0.8

250

0.9

1.0

0.8

0.6

1.5

1.2

1.0

500

0.7

0.8

1.0

0.2

1.0

0.7

1.3

1000

0.7

0.7

1.0

0.4

0.7

0.2

1.4

2000

1.1

0.9

1.1

0

0.4

0

1.2

4000

1.0

1.0

1.0

0

0.7

0

1.1

8000

0.8

1.0

0.5

0

0.7

0

1.1

Sodium azide 2 µg

-

-

15.2

-

-

-

-

Benzo(a)-pyrene 7 µg

-

-

-

-

3.7

5.3

3.1

4-nitroquinoline-N-oxide 3 µg

1.2

3.6

-

-

-

-

-

Neutral red 7 µg

-

-

-

4.4

-

-

-

Table: Relative Mutation Rate – Bacterial Experiment II without Metabolic Activation

Concentration

E. coli WP2

E. coli WP2 uvr A

S. typhimurium TA 1535

S. typhimurium TA 1537

S. typhimurium TA 1538

S. typhimurium TA 98

S. typhimurium TA 100

Without S9

3.91

-

-

-

1.1

-

0.8

-

7.81

-

-

-

0.6

-

0.9

-

15.6

-

-

-

0.7

0.9

0.8

-

31.3

-

-

-

0.4

1.0

0.8

1.1

62.5

1.0

1.0

1.2

0.4

0.7

0.4

1.0

125

1.0

1.0

0.9

0

0.5

0

0.9

250

0.7

0.8

1.4

-

0.2

-

1.0

500

0.9

0.7

1.1

-

0

-

0.7

1000

1.0

0.8

0.9

-

0

-

-

2000

0.7

0.9

0.7

-

-

-

-

4000

0.9

0.8

0.9

-

-

-

-

8000

0.8

0.7

0.9

-

-

-

-

Sodium azide 2 µg

-

-

86.6

-

-

-

-

Benzo(a)-pyrene 7 µg

-

-

-

-

1.1

1.1

1.3

4-nitroquinoline-N-oxide 3 µg

36.7

5.3

-

-

-

-

-

Neutral red 7 µg

-

-

-

1.2

-

-

-

Table: Relative Mutation Rate – Bacterial Experiment II with Metabolic Activation

Concentration

E. coli WP2

E. coli WP2 uvr A

S. typhimurium TA 1535

S. typhimurium TA 1537

S. typhimurium TA 1538

S. typhimurium TA 98

S. typhimurium TA 100

Without S9

31.3

-

-

-

1.1

0.9

0.8

-

62.5

0.9

0.9

0.7

1.2

1.1

1.0

1.1

125

1.1

1.0

1.2

1.1

1.1

0.9

1.0

250

1.4

0.8

0.8

1.4

1.1

0.8

1.1

500

1.2

1.0

0.9

1.0

1.1

0.8

1.2

1000

1.0

1.1

0.8

0.5

0.9

0.4

0.9

2000

0.8

1.0

1.1

0

0.9

0.1

0.9

4000

0.8

0.9

0.8

-

0.5

-

0.9

8000

1.0

0.7

0.7

-

0.4

-

0.9

Sodium azide 2 µg

-

-

18.2

-

-

-

-

Benzo(a)-pyrene 7 µg

-

-

-

-

9.7

6.1

6.5

4-nitroquinoline-N-oxide 3 µg

1.1

2.8

-

-

-

-

-

Neutral red 7 µg

-

-

-

5.2

-

-

-

Applicant's summary and conclusion

Conclusions:
Interpretation of results:
negative with metabolic activation
negative without metabolic activation

The purpose of this study was to determine the mutagenicity of the test substance hydrocarbons, C7-C9, n-alkanes, isoalkanes, cyclics. An Ames reverse mutation assay was done on S. typhimurium strains, TA 1535, TA 1537, TA 1538, TA 98 and TA 100, and E. coli strains WP2 uvr A and WP2, both with and without metabolic activation. Species were tested at concentrations ranging from 0.31 to 8000 ug/plate. The test plates were incubated for 48 -72 hrs, and then the number of colonies were counted. For all species, there was no significant increase in the number of revertants as compared to negative controls. Positive controls showed a significant increase in mutations. The test substance is not mutagenic.
Executive summary:

The purpose of this study was to determine the mutagenicity of the test substance hydrocarbons, C7-C9, n-alkanes, isoalkanes, cyclics. An Ames reverse mutation assay was done on S. typhimurium strains, TA 1535, TA 1537, TA 1538, TA 98 and TA 100, and E. coli strains WP2 uvr A and WP2, both with and without metabolic activation. Species were tested at concentrations ranging from 0.31 to 8000 ug/plate. The test plates were incubated for 48 -72 hrs, and then the number of colonies were counted. For all species, there was no significant increase in the number of revertants as compared to negative controls. Positive controls showed a significant increase in mutations. The test substance is not mutagenic.