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Diss Factsheets

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:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data from handbook or collection of data

Data source

Reference
Reference Type:
publication
Title:
Chemical Mutagenesis at the Thymidine Kinase Locus in C5178Y Mouse Lv;nPhoma #B Cells: Results for 31 Coded Compounds in the National Toxicology Program
Author:
Mhyr and Caspary
Year:
1991
Bibliographic source:
Environmental and Molecular Mutagenesis 1861-83

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Principles of method if other than guideline:
Equivalent or similar to OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
GLP compliance:
not specified
Type of assay:
in vitro mammalian cell gene mutation tests using the thymidine kinase gene

Test material

Constituent 1
Chemical structure
Reference substance name:
Dodecan-1-ol, ethoxylated
EC Number:
500-002-6
EC Name:
Dodecan-1-ol, ethoxylated
Cas Number:
9002-92-0
Molecular formula:
C58H118024
IUPAC Name:
Dodecan-1-ol, ethoxylated
Test material form:
liquid
Details on test material:
Name: Dodecan-1-ol, ethoxylated
CAS No. 9002-92-0
Molecular Weight: 230.389 g/mol
Molecular Formula: (C2-H4-O)mult-C12-H26-O

Method

Target gene:
Thymidine Kinase
Species / strain
Species / strain / cell type:
mouse lymphoma L5178Y cells
Details on mammalian cell type (if applicable):
No Data Available
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
not specified
Metabolic activation:
with and without
Metabolic activation system:
Type and composition of metabolic activation system:
- source of S9 : Freshly prepared S9 from the livers of either Aroclor 1254-induced or non-induced male Fischer 344 rats.
Test concentrations with justification for top dose:
0,5,19,15,20,30,30,40,50 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Water
- Justification for choice of solvent/vehicle: The test substance is soluble in water
Controlsopen allclose all
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
Water
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
Water
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
3-methylcholanthrene
Details on test system and experimental conditions:
NUMBER OF REPLICATIONS:
- Number of cultures per concentration: Triplicate
- Number of independent experiments

METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable):
- Test substance added in medium

TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable: No Data Available
- Exposure duration/duration of treatment: 4 hours
- Harvest time after the end of treatment (sampling/recovery times): No Data Available


FOR GENE MUTATION:
- Expression time (cells in growth medium between treatment and selection): 48 hours
- Selection time (if incubation with a selective agent): 10 to 12 days
- Fixation time (start of exposure up to fixation or harvest of cells): After the 48-hour expression period, 3 x 10^6 cells were plated in medium and soft agar supplemented with trifluorothymidine (TFT) for selection of TFT-resistant cells (TK-/-) and in nonselective medium and soft agar to determine cloning efficiency. Plates were incubated at 37 C. in 5% CO2 for 10 to 12 days.
- Method used: Microwell plates for the mouse lymphoma assay.
- If a selective agent is used: trifluorothymidine was used
- Number of cells seeded and method to enumerate numbers of viable and mutants cells: 3 x 10^6 cells
- Criteria for small (slow growing) and large (fast growing) colonies: No Data Available

METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method: Relative total growth (RTG)
- Any supplementary information relevant to cytotoxicity: No Data Available

METHODS FOR MEASUREMENTS OF GENOTOXICITY: No Data Available

- OTHER: No Data Available
Rationale for test conditions:
No Data Available
Evaluation criteria:
The mammalian cells were observed for mutagenic frequency in cells.
Statistics:
Yes, SD ± Mean was observed.

Results and discussion

Test results
Key result
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
not specified
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
No Data Available
Remarks on result:
other: No mutagenic effect were observed

Any other information on results incl. tables

Nonactivation Trial:2 Experiment Call:Negative

 

Conc.
µg/mL

Cloning
Efficiency

Relative Total
Growth

Mutant Colonies

Mutant Frequency

AVG Mutant Frequency

Vehicle Control

Water

0         

110

113

128

39

33

 

 

108

83

114

35

 

 

116

124

84

24

 

 

108

80

115

36

Test Chemical

Test chemical

5         

106

107

113

35

30

 

 

113

109

97

29

 

 

115

119

93

27

10         

105

80

139

44

40

 

 

107

97

171

53

 

 

112

117

78

23

15         

105

107

136

43

41

 

 

114

82

144

42

 

 

115

71

125

36

20         

104

72

218

70

48

 

 

105

74

101

32

 

 

117

79

149

42

30         

107

29

114

35

51*

 

 

110

34

208

63

 

 

108

29

180

56

40         

LETHAL

 

 

 

LETHAL

 

 

LETHAL

Positive Control

Methyl Methane Sulfonate

5         

105

60

1132

360

392*

 

 

115

56

1120

326

 

 

86

23

1257

489

Induced S9 Trial:2Experiment Call:Negative

 

Conc.
µg/mL

Cloning
Efficiency

Relative Total
Growth

Mutant Colonies

Mutant Frequency

AVG Mutant Frequency

Vehicle Control

Water

0         

112

65

106

32

31

 

 

113

94

107

32

 

 

105

144

81

26

 

 

116

98

129

37

Test Chemical

Test chemical

5         

97

96

112

39

33

 

 

106

108

92

29

 

 

106

95

102

32

10         

105

100

91

29

30

 

 

100

78

90

30

 

 

116

98

112

32

15         

97

56

118

41

39

 

 

117

70

121

35

 

 

113

61

144

42

20         

105

76

129

41

30

 

 

121r

72

85

23

 

 

112

51

67

20

30         

101

32

120

40

40

 

 

105

43

139

44

 

 

111

46

117

35

40         

104

13

84

27

39

 

 

107

32

164

51

 

 

LETHAL

50         

LETHAL

 

 

 

LETHAL

 

 

LETHAL

Positive Control

Methyl Methane Sulfonate

5         

111

43

630

189

360*

 

 

78

31

1010

432

 

 

47

12

646

460

Applicant's summary and conclusion

Conclusions:
Based on all the observations and results, it was concluded that the test chemical was non-mutagenic in nature and did not cause any genotoxicity at the tested dose levels.
Executive summary:

An in vitro mammalian cell gene mutation assay using the mouse lymphoma L5178Y cells was performed to observe the genotoxic potential of the test chemical. In this study, the average mutant frequency was observed on the Thymidine Kinase gene locus. The study was performed using with and without metabolic activation. Freshly prepared S9 from the livers of either Aroclor 1254-induced or non-induced male Fischer 344 rats. The concentration of test material used in two experiment were 0,5,19,15,20,30,30,40,50 µg/mL. The test chemical was dissolved in Water which was consider as a vehicle control. Methyl methanesulphonate was used a positive control. The average mutant frequency was considered as a measure for mutagenicity. However, the test chemical did not induce mutagenicity at the gene locus and therefore was considered to be non-mutagenic in mouse lymphoma cells by in vitro mammalian cell gene mutation assay. Therefore, based on all the observations and results, it was concluded that the test chemical was non-mutagenic in nature and did not cause any genotoxicity at the tested dose levels.