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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:
2006-09-20 until 2006-10-13
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report date:
2006

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
not specified
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
2-butyloctan-1-ol
EC Number:
223-470-0
EC Name:
2-butyloctan-1-ol
Cas Number:
3913-02-8
Molecular formula:
C12H26O
IUPAC Name:
2-butyloctan-1-ol
Details on test material:
- Name of test material (as cited in study report): Isofol 12
- Substance type: not mentioned
- Physical state: liquid
- Lot/batch No.: A68263
- Expiration date of the lot/batch: 2010-03-29
- Stability under test conditions: stable under storage conditions
- Storage condition of test material: at room temperature

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
liver S9 mix of Phenobarbital/ß-Naphthoflavone induced rats
Test concentrations with justification for top dose:
Experiment I: 3, 10, 33, 100, 333, 1000, 2500 and 5000 µg/plate; Experiment II: 10, 33, 100, 333, 1000, 2500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: not mentioned
Controlsopen allclose all
Untreated negative controls:
yes
Remarks:
Untreated
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
methylmethanesulfonate
other: 4-Nitro-o-phenylene-diamine (10 µg 4-NOPD/plate in TA 98 and 50 µg 4-NOPD/plate in TA 1537
Remarks:
without metabolic activation
Untreated negative controls:
yes
Remarks:
Untreated
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene (2.5 µg 2-AA/plate, TA 1535, TA 1537, TA 98 and TA 100), 10 µg 2-AA/plate in WP2 uvrA
Remarks:
with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: The study was performed in two independent tests in the plate incorporation test (Experiment I) and the pre-incubation test (Experiment II)

DURATION
- Preincubation period: 60 minutes
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 3 per concentration

DETERMINATION OF CYTOTOXICITY
- Method: not mentioned

OTHER EXAMINATIONS:
- Other: Determination of the frequency of induced or spontaneous reversion to histidine independence with negative controls (H2O), solvent controls (DMSO), test substance concentrations and positive controls; determination of the titers of overnight cultures

OTHER: none
Evaluation criteria:
The test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA 98, TA 100, and WP2 uvrA) or thrice (strains TA 1535 and TA 1537) the colony count of the corresponding solvent control is observed. A dose dependent increase is considered biologically relevant if the threshold is exceeded at more than one concentration. An increase exceeding the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment. A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls such an increase is not considered biologically relevant.
Statistics:
no statistics performed

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: not measured
- Effects of osmolality: not applicable
- Evaporation from medium: not applicable
- Water solubility: not mentioned
- Precipitation: no
- Other confounding effects: none

RANGE-FINDING/SCREENING STUDIES: not performed

COMPARISON WITH HISTORICAL CONTROL DATA: not performed

ADDITIONAL INFORMATION ON CYTOTOXICITY: none

Any other information on results incl. tables

Table #1: Pre-Experiment/Experiment I: Number of revertants per plate (mean of 3 plates)

 

[Strain TA 98]

[Strain TA 100]

[Strain TA 1535]

Conc.
[µg/plate]

- MA

+ MA

Cytotoxic
(yes/no)

- MA

+ MA

Cytotoxic
(yes/no)

- MA

+ MA

Cytotoxic
(yes/no)

0*

29 ± 1

44 ± 4

no

164 ± 24

195 ± 32

no

24 ± 4

 33 ± 11

no

3

35 ± 5

29 ± 6

no

186 ± 12

186 ± 24

no

29 ± 3

32 ± 9

no

10

34 ± 7

41 ± 10

no

150 ± 10

191 ± 10

no

27 ± 5

30 ± 7

no

33

30 ± 8

40 ± 5

no

129 ± 25

207 ± 31

no

24 ± 13

26 ± 2

no

100

26 ± 1

31 ± 2

no

128 ± 15

191 ± 13

no

22 ± 2

24 ± 8

no

333

20 ± 3

31 ± 7

no

140 ± 16

145 ± 11

no

24 ± 6

19 ± 4

no

1000

22 ± 6

31 ± 1

no

131 ± 17

110 ± 7

no

27 ± 7

18 ± 4

no

 2500 21 ± 6 5 ± 2 R no 129 ± 20 61 ± 6 R no 14 ± 5 21 ± 4 R no
 5000 23 ± 4  15 ± 1 R no 82 ± 6 51 ± 3 R no 19 ± 2 23 ± 2 R no
 Positive Control 489 ± 35 2581 ± 503 no 2170 ± 80 4310 ± 73 no 1460 ± 14 524 ± 28 no

*solvent control with DMSO; R = Reduced background growth

Table #2: Pre-Experiment/Experiment I: Number of revertants per plate (mean of 3 plates)

 

[Strain TA 1537]

[WP2 uvrA]

Conc.
[µg/plate]

- MA

+ MA

Cytotoxic
(yes/no)

- MA

+ MA

Cytotoxic
(yes/no)

0*

15 ± 2

16 ± 1

no

57 ± 7

73 ± 4

no

3

15 ± 2

16 ± 7

no

53 ± 3

85 ± 19

no

10

17 ± 3

24 ± 9

no

58 ± 11

79 ± 5

no

33

11 ± 5

21 ± 1

no

55 ± 5

70 ± 3

no

100

13 ± 3

14 ± 5

no

56 ± 3

65 ± 2

no

333

11 ± 1

18 ± 7

no

75 ± 20

88 ± 14

no

1000

7 ± 4

11 ± 3

no

62 ± 9

85 ± 16

no

2500 5 ± 3 5 ± 2 R no 48 ± 5 40 ± 17 R no
5000 6 ± 1 0 ± 0 R no 54 ± 8 31 ± 7 R no
 Positive Control 103 ± 30 462 ± 3 no 1085 ± 102 251 ± 42 no

*solvent control with DMSO; R = Reduced background growth

Table #3: Experiment II: Number of revertants per plate (mean of 3 plates)

 

[Strain TA 98]

[Strain TA 100]

[Strain TA 1535]

Conc.
[µg/plate]

- MA

+ MA

Cytotoxic
(yes/no)

- MA

+ MA

Cytotoxic
(yes/no)

- MA

+ MA

Cytotoxic
(yes/no)

0*

35 ± 1

33 ± 7

no

135 ± 2

157 ± 2

no

16 ± 5

19 ± 4

no

10

31 ± 3

38 ± 5

no

121 ± 12

138 ± 5

no

15 ± 1

16 ± 6

no

33

31 ± 2

38 ± 3

no

93 ± 5

147 ± 9

no

9 ± 1

27 ± 4

no

100

25 ± 4

28 ± 3

no

101 ± 3

99 ± 10

no

6 ± 2

17 ± 6

no

333

21 ± 4

25 ± 9

no

81 ± 17

98 ± 21

no

13 ± 4

10 ± 2

no

1000

10 ± 3

23 ± 7

no

52 ± 11

64 ± 8

no

17 ± 4

10 ± 2

no

2500 12 ± 2 MR 21 ± 4 MR no 42 ± 4 MR 39 ± 4 MR no 12 ± 3 MR 7 ± 2 MR no
5000 6 ± 3 MR  11 ± 3 MR no 13 ± 8 MR 16 ± 2 MR no 7 ± 3 MR 5 ± 1 MR no
 Positive Control 500 ± 35 1492 ± 100 no 1863 ± 59 2839 ± 96 no 1656 ± 61 284 ± 15 no

*solvent control with DMSO; M = Manual count; R = Reduced background growth

Table #4: Experiment II: Number of revertants per plate (mean of 3 plates)

 

[Strain TA 1537]

[WP2 uvrA]

Conc.
[µg/plate]

- MA

+ MA

Cytotoxic
(yes/no)

- MA

+ MA

Cytotoxic
(yes/no)

0*

10 ± 4

15 ± 2

no

49 ± 12

53 ± 5

no

10

7 ± 6

20 ± 4

no

51 ± 4

60 ± 4

no

33

10 ± 3

17 ± 4

no

45 ± 4

49 ± 13

no

100

17 ± 10

6 ± 0

no

44 ± 4

55 ± 6

no

333

12 ± 2

10 ± 2

no

47 ± 4

49 ± 20

no

1000

7 ± 2

9 ± 3

no

51 ± 8

52 ± 11

no

2500 6 ± 2 MR 1 ± 1 MR no 35 ± 7 MR 31 ± 3 MR no
5000 2 ± 1 MR 0 ± 0 MR no 28 ± 4 MR 26 ± 6 MR no
 Positive Control 119 ± 13 319 ± 44 no 752 ± 55 208 ± 31 no

*solvent control with DMSO; M = Manual Count; R = Reduced background growth

 

Applicant's summary and conclusion

Conclusions:
Based on the results of this study under the experimental conditions 2-butyloctan-1-ol does not induce gene mutations by base pair
changes or frameshifts in the genome and is therefore not mutagenic.
Executive summary:

Based on the results of this study under the experimental conditions 2-butyloctan-1-ol does not induce gene mutations by base pair

changes or frameshifts in the genome and is therefore not mutagenic.