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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
19-Sep-1996 to 24-Sep-1996
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
The restriction was that no cross linking strain was used, so does not comply with current guidelines.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report Date:
1996

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
(no TA102 or E coli WP2 uvrA, 2-AA only positive control with S9)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid

Method

Target gene:
histidine
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 1538
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced rat liver S9
Test concentrations with justification for top dose:
Test 1: 15(-S9 only), 50, 150, 500, 1500, and 5000 µg/plate; Test 2: 50, 150, 500, 1500, and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Well-known solvent
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Remarks:
TA100 (3 ug/plate) and TA1535 (5 ug/plate) without S9
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Remarks:
TA1537 (8 ug/plate) without S9
Positive control substance:
9-aminoacridine
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Remarks:
TA1538 (5 ug/plate) without S9
Positive control substance:
other: 4-nitro-o-phenylenediamine
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Remarks:
TA98 (0.2 ug/plate) without S9
Positive control substance:
4-nitroquinoline-N-oxide
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Remarks:
all strains (0.5, 1 or 2 ug/plate) with S9
Positive control substance:
other: 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium; in agar (plate incorporation)

DURATION
- Exposure duration: approximately 48 hours

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Thinning or absence of background lawn of non-revertant cells

OTHER: The mutation experiment was repeated on a separate day using fresh cultures and fresh test material formulations.
Evaluation criteria:
After incubation, numbers of revertant colonies were counted. For a substance to be considered positive in this test system, it should have induced a dose-related and statistically significant increase in mutation rate in one or more strains of bacteria in the presence and/or absence of S9 in both experiments at sub-toxic dose levels. For a negative result the numbers of induced revertants should be less than two fold compared to controls.
Statistics:
Dunnetts test was used and showed no statistically significant differences between test and control plates.

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no data
- Effects of osmolality: no data
- Evaporation from medium: no data
- Water solubility: no data
- Precipitation: precipitation of test substance was observed at dose levels of 1500 ug/plate and above. Plates were counted manually at 1500 ug/plate and above.
- Other confounding effects: no data

RANGE-FINDING/SCREENING STUDIES: Strain TA 100 was exposed to the test substance at concentrations of 0, 50, 150, 500, 1500 and 5000 ug/plate in a preliminary toxicity study.

COMPARISON WITH HISTORICAL CONTROL DATA: no data

CYTOTOXIC CONCENTRATION: Slight cytotoxicity was indicated in a preliminary toxicity screen with TA100 at dose levels >= 500 ug/plate without 
metabolic activation. In the actual mutation study there was no evidence of cytotoxicity up to 5000 ug/plate with or without S9.
Remarks on result:
other: No mutagenic potential

Any other information on results incl. tables

Table 1:Number of revertants per plate (mean of 3 plates) for Test 1

 

Conc.
[
µg /plate]

 

[TA 100]

[TA 1535]

[TA 1538]

[TA 98]

[TA 1537]

-MA

+MA

Cytotoxic
(yes/no)

- MA

+MA

Cytotoxic
(yes/no)

- MA

+MA

Cytotoxic
(yes/no)

-MA

+ MA

Cytotoxic
(yes/no)

-MA

+ MA

Cytotoxic
(yes/no)

0*

72

116

no

14

15

no

14

25

no

20

29

no

6

9

no

15

71

-

no

12

-

no

11

-

no

18

-

no

7

-

no

50

65

112

no

12

15

no

11

27

no

17

24

no

6

12

no

150

64

98

no

12

12

no

12

22

no

18

35

no

7

12

no

500

57

89

no

12

14

no

11

28

no

21

27

no

8

10

no

1500

55

72

no

12

17

no

11

22

no

19

27

no

7

9

no

5000

52

74

no

12

11

no

12

22

no

23

30

no

10

7

no

Positive control

597

831

no

348

195

no

636

488

no

217

485

no

550

282

no

*solvent control with DMSO

 

Table 2: Number of revertants per plate (mean of 3 plates) for Test 2

 

Conc.
[
µg /plate]

 

[TA 100]

[TA 1535]

[TA 1538]

[TA 98]

[TA 1537]

-MA

+MA

Cytotoxic
(yes/no)

-MA

+MA

Cytotoxic
(yes/no)

-MA

+MA

Cytotoxic
(yes/no)

-MA

+MA

Cytotoxic
(yes/no)

-MA

+MA

Cytotoxic
(yes/no)

0*

86

85

no

19

12

no

11

16

no

15

24

no

7

9

no

50

67

83

no

15

13

no

10

14

no

19

28

no

7

8

no

150

70

66

no

11

9

no

9

13

no

20

29

no

5

7

no

500

66

71

no

17

10

no

9

14

no

15

26

no

6

11

no

1500

66

72

no

9

11

no

7

12

no

13

20

no

5

8

no

5000

66

58

no

9

10

no

6

13

no

12

23

no

7

9

no

Positive control

459

1005

no

211

208

no

524

344

no

175

345

no

762

266

no

*solvent control with DMSO

 

 



Applicant's summary and conclusion

Conclusions:
In a reliable study, conducted in accordance with OECD guideline 471 and under GLP, the C14 alcohol Kalcol 4098 did not increase the reverse mutation rate in histidine dependent bacterial strains of Salmonella typhimurium in the presence or absence of metabolic activation at concentrations up to 5000 µg/plate. The top concentration was not cytotoxic. It is concluded that the test substance is not mutagenic to bacteria under the conditions of the test.
Executive summary:

In the bacterial mutagenicity study, the ability of tetradecan-1 -ol to induce mutations in bacteria was tested.

In test I, 50, 150, 500, 1500, and 5000 µg/plate of test material were applied to S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 1538 without metabolic activation. In test II, the same concentrations of the test material were applied to

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 1538, in the presence and absence of metabolic activation system. The exposure duration was approximately 48 hours.

After incubation, numbers of revertant colonies were counted. For a substance to be considered positive in this test system, it should have induced a dose-related and statistically significant increase in mutation rate in one or more strains of bacteria in the presence and/or absence of metabolic activation in both experiments at sub-toxic dose levels. For a negative result the numbers of induced revertants should be less than two fold compared to controls.

The study reports tetradecan-1 -ol to be not mutagenic to S. typhimurium when tested up to limit concentration. The study was conducted according to an appropriate OECD test guideline, with acceptable restriction. The restriction was that no cross linking strain was used, so does not comply with current guidelines. It was compliant with GLP.