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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in mammalian cells
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:
1983-01-21 to 1983-06-24
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study is classified as reliable without restrictions because it was carried out in a method equivalent/similar to OECD TG 476.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1984
Report Date:
1984

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
GLP compliance:
yes
Type of assay:
mammalian cell gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: low viscosity liquid hydrocarbon
Details on test material:
Test substance: Hydrodesulfurised kerosine Sample API 81-07, CAS No. 64742-81-0

Method

Species / strain
Species / strain / cell type:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Test concentrations with justification for top dose:
3.91 to 6.25 nl/ml with activation, 6.25 to 37.5 nl/ml without activation.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: none
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
corn oil
Positive controls:
yes
Positive control substance:
triethylenemelamine
Details on test system and experimental conditions:
Mouse lymphoma assay

Results and discussion

Test results
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
The test material induced a good range of toxicities for evaluation (relative growths ranged from 2.8% to 65.3%, -S9, and 6.1 to 107.9% relative growths, +S9)
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'. Remarks: mouse lymphoma L5178Y cells

Any other information on results incl. tables

The test material was immiscible with water and DMSO at 100 µl/ml but was miscible with ethanol at the same concentration. Stocks were prepared by performing serial dilutions in ethanol just prior to each assay. The mutation assays were then initiated by performing final dilutions of the stocks into the assay medium containing the lymphoma cells. The test material appeared soluble in the assay medium up to 125 nl/ml but a white precipitate was noted from 250 to 1000 nl/ml.

Two trials of the assay were initiated.
Trial 1 was performed with and without activation but the non-activation assay was NOT used in the evaluation because of unacceptable suspension growth in the negative controls. The non-activation portion of the assay was therefore repeated in Trial 2.

The report summarized here included the acceptable activation assay from Trial 1 and the acceptable non-activation assay from Trial 2.

The results are summarized below.
 

 

Rel

Total

Total

Cloning

Rel

Mutant

 

Susp.

mutant

viable

eff.

growth

frequency

 

growth

  colonies

 

(%)

10E-6units

 

 

 (% of control)

 

 

 

 

 

Non activation assay (Trial 2)

Solvent control (ethanol)

 

100

70

392

100

100

17.9

 

100

74

287

100

100

25.8

 

100

75

367

100

100

20.4

 

100

50

324

100

100

15.4

Untreated control

 

107.1

82

374

109.2

116.7

21.9

 

 97.4

70

392

116.5

111.3

17.9

EMS(µl/ml)

0.5

46.7

851

170

49.6

23.2

500.6

0.5

57.3

753

155

45.3

25.9

485.8

API81-07 (nl/ml)

 6.25

28.3

83

331

96.6

27.3

25.1

12.5

24.7

61

282

82.3

20.3

21.6

12.5

67.4

89

332

96.9

65.3

26.8

25.0

16.3

87

258

75.3

12.3

33.7

25.0

45.4

61

248

72.4

82.7

24.6

37.5

10.3

105

366

106.8

11

28.7

37.5

6.2

77

143

41.7

2.6

53.8

Activation assay

Solvent control (ethanol)

 

100

98

236

100

100

41.5

 

100

92

282

100

100

32.6

Untreated control

 

128.2

99

226

87.2

111.8

43.8

DMN( µl/ml)

 0.3

58.5

151

34

13.1

7.7

444.1

API81-07 (nl/ml)

 3.91

100.1

66

194

74.8

74.9

34

 7.81

194.1

52

144

55.6

107.9

36.1

15.6

101.7

42

259

99.9

101.6

16.2

31.3

 27.3

45

158

61

16.7

28.5

62.5

 9

68

175

67.5

6.1

38.9

 
Under non-activation conditions the test material induced a good range of toxicities for evaluation (relative growths ranged from 2.8% to 65.3%).
None of the assays induced a mutant frequency that exceeded the minimum criterion (40.8 x 10-6). The test material was not mutagenic under non-activation conditions.

In the presence of metabolic activation, a wide range of toxicities were induced (6.1 to 107.9% relative growths). The minimum criterion mutant frequency of 69.0 x 10-6 was not exceeded. The test material was therefore considered non mutagenic under activation conditions.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

The test substance does not induce chromosomal aberrations in rat bone marrow cells under the condition of the study. ... (see attached file)
Executive summary:

In a mammalian cell gene mutation assay, mouse lymphoma L5178Y cells, cultured in vitro, were exposed to hydrodesulfurised kerosine at concentrations from 6.25 nL/mL to 37.5 nL/mL without activation and from 3.91 nl/ml to 62.5 nl/ml with S9 metabolic activation. The cells were exposed for four hours both in the presence and absence of the rat liver S9 metabolic activation.

Under non-activation conditions the test material induced a good range of toxicities for evaluation (relative growths ranged from 2.8% to 65.3%). None of the assays induced a mutant frequency that exceeded the minimum criterion (40.8 x 10-6). The test material was not mutagenic under non-activation conditions. In the presence of metabolic activation, a wide range of toxicities were induced (6.1 to 107.9% relative growths). The minimum criterion mutant frequency of 69.0 x 10-6 was not exceeded. The test material was therefore considered non mutagenic under activation conditions.

This study received a Klimisch score of 1 and is classified as reliable without restriction because it was carried out in a method similar/equivalent to OECD TG 471.