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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Test protocol comparable to OECD Guideline 471 with acceptable restrictions
Cross-reference
Reason / purpose:
reference to other study

Data source

Referenceopen allclose all

Reference Type:
other: Database entry
Title:
Unnamed
Year:
1982
Reference Type:
study report
Title:
Unnamed
Year:
2003
Reference Type:
publication
Title:
Salmonella Mutagenicity Tests: III . Results From the Testing of 255 Chemicals
Author:
Zeiger E, Anderson B, Haworth S, Lawlor T, Mortelmans K, Speck W
Year:
1987
Bibliographic source:
Environ. Mutagen. 9, Suppl. 9, 1-110
Reference Type:
other: abstract
Title:
TR-505 Toxicology and Carcinogenesis Studies of Citral (Microencapsulated) (CAS No. 5392-40-5) in F344/N Rats and B6C3F1 Mice (Feed Studies) - Draft Abstract
Author:
National Toxicology Program NTP
Year:
2003
Bibliographic source:
http://ntp-server.niehs .nih .gov/htdocs/LT-studies/tr505 .html

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
(four tested strains: TA1535, TA1537, TA98, TA100)
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Citral
- Analytical purity: 89% (data from Zeiger et al., 1987)

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
rat and hamster liver S9 (Aroclor induced)
Test concentrations with justification for top dose:
-S9: 1-67 µg/plate
+S9: 3.3-220 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: see remarks
Remarks:
without metabolic activation: TA98, 2-nitrofluorene; TA100 and TA1535, sodium azide; TA1537, 9-aminoacridine; with metabolic activation: all strains, 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 2 days

NUMBER OF REPLICATIONS: 3 per trial, 2 trials

DETERMINATION OF CYTOTOXICITY
- Method: decrease in number of his+ colonies or clearing in the density of background lawn
Evaluation criteria:
mutagenic response:
Individual trial: dose-related increase above solvent control
all trials combined: reproducible, dose-related increase over solvent control in replicate trials

Results and discussion

Test results
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:
cytotoxicity
Remarks:
without S9-mix: 67 µg/plate, with S9-mix: 160 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Mutagenicity of citral in Salmonella typhimurium

Strain TA 1535

Dose

No Activation
(Trial 1)

No Activation
(Trial 2)

10% HLI
(Trial 1)

10% HLI
(Trial 2)

10% RLI
(Trial 1)

10% RLI
(
Trial 2)

µg/plate

Mean

± SEM

Mean

± SEM

Mean

± SEM

Mean

± SEM

Mean

± SEM

Mean

± SEM

0         

29

4.5

25

2.6

17

2.6

9

1.7

14

1.5

7

1.9

1         

25

1.9

18

3

 

 

 

 

 

 

 

 

3.3       

27

3.8

23

1.7

14

2

11

1.2

17

0.6

12

0.9

10         

27

3.8

23

4.1

13

2.9

12

3.8

8

0.3

9

2.1

33         

26

3.8

23

4.3

16

2.3

12

0.3

13

1.5

13

3.8

50         

 

 

16S

2

 

 

 

 

 

 

 

 

67         

19S

2.3

 

 

 

 

 

 

 

 

 

 

100         

 

 

 

 

18

3

14

2.7

11

1.2

8

0.6

160         

 

 

 

 

 

 

11S

3.7

 

 

10S

1.3

220         

 

 

 

 

T

 

 

T

 

 

Trial summary

Negative

Negative

Negative

Negative

Negative

Negative

Positive Control

1277

17.6

1098

39.7

119

5.2

57

2.8

99

6.2

71

5.3

Strain TA 100

Dose

No Activation
(Trial 1)

No Activation
(Trial 2)

10% HLI
(Trial 1)

10% HLI
(Trial 2)

10% RLI
(Trial 1)

10% RLI
(
Trial 2)

µg/plate

Mean

± SEM

Mean

± SEM

Mean

± SEM

Mean

± SEM

Mean

± SEM

Mean

± SEM

0         

155

3.2

154

6.1

148

9

146

5.8

122

7.9

150

8.4

1         

152

16.8

146

9.4

 

 

 

 

 

 

 

 

3.3       

136

7.2

141

2.4

157

4.8

149

2.2

111

11.3

128

3.5

10         

140

6.9

143

7.1

145

9.7

134

8.4

126

2.8

122

1.7

33         

134

3.8

147

14

145

0.9

142

5.2

123

8.1

138

2.1

50         

 

 

132S

12.9

 

 

 

 

 

 

 

 

67         

T

 

 

 

 

 

 

 

 

 

 

100         

 

 

 

 

151

5.9

142

6.3

129

1

150

9.4

160         

 

 

 

 

 

 

141S

1.9

 

 

126T

5.5

220         

 

 

 

 

T

 

 

T

 

 

Trial summary

Negative

Negative

Negative

Negative

Negative

Negative

Positive Control

1386

11.9

1349

18.9

900

38.1

439

24.6

774

9.1

434

32.4

Strain TA 98

Dose

No Activation
(Trial 1)

No Activation
(Trial 2)

10% HLI
(Trial 1)

10% HLI
(Trial 2)

10% RLI
(Trial 1)

10% RLI
(
Trial 2)

µg/plate

Mean

± SEM

Mean

± SEM

Mean

± SEM

Mean

± SEM

Mean

± SEM

Mean

± SEM

0         

33

2.6

15

1.8

40

1.5

24

2

29

1.5

23

0.9

1         

28

1

18

2.3

 

 

 

 

 

 

 

 

3.3       

25

2.2

13

2.8

32

4.1

25

4.4

34

2.7

20

3.6

10         

26

5.4

17

1.8

38

3.4

28

4.6

34

2.5

22

0.9

33         

30

2.8

16

0.6

40

5.5

25

3.3

31

6.2

21

1.2

50         

 

 

13

1.2

 

 

 

 

 

 

 

 

67         

23S

0.9

 

 

 

 

 

 

 

 

 

 

100         

 

 

 

 

30

1.5

29

4.1

34

3

21

1

160         

 

 

 

 

 

 

21S

3.4

 

 

23S

1.5

220         

 

 

 

 

T

 

 

T

 

 

Trial summary

Negative

Negative

Negative

Negative

Negative

Negative

Positive Control

1510

60.1

1171

55

890

39.3

288

6.8

679

19.5

328

20.3

Strain TA 1537

Dose

No Activation
(Trial 1)

No Activation
(Trial 2)

10% HLI
(Trial 1)

10% HLI
(Trial 2)

10% RLI
(Trial 1)

10% RLI
(
Trial 2)

µg/plate

Mean

± SEM

Mean

± SEM

Mean

± SEM

Mean

± SEM

Mean

± SEM

Mean

± SEM

0         

10

0.3

5

0.9

10

3.2

7

0.9

9

1.5

7

1

1         

9

1.5

5

0.7

 

 

 

 

 

 

 

 

3.3       

9

0.9

6

1.7

10

3.7

3

0.7

9

1.2

6

1

10         

8

1.7

6

0.9

13

2.8

8

1

8

0.9

7

1.2

33         

6

1.3

7

0.3

12

2

5

0.9

6

0.6

5

1

50         

 

 

3S

1.2

 

 

 

 

 

 

 

 

67         

T

 

 

 

 

 

 

 

 

 

 

100         

 

 

 

 

9

0.6

6

1.5

9

0.7

7

1.9

160         

 

 

 

 

 

 

5S

0.7

 

 

7S

0.6

220         

 

 

 

 

T

 

 

T

 

 

Trial summary

Negative

Negative

Negative

Negative

Negative

Negative

Positive Control

547

36.5

516

58.9

74

1.2

30

0.3

54

3.2

34

6.4

Abbreviations:
RLI = induced male Sprague Dawley rat liver S9
HLI = induced male Syrian hamster liver S9
s = Slight Toxicity; p = Precipitate; x = Slight Toxicity and Precipitate; T = Toxic; c = Contamination

Applicant's summary and conclusion