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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: The purity of the test substance was known. The authors cited the Ames et al. (1975) methods paper with a detailed protocol following the cited procedure. The results were described in details.

Data source

Reference
Reference Type:
publication
Title:
Mutagenicity testing in the Salmonella typhimurium assay of phenolic compounds and phenolic fractions obtained from smokehouse smoke condensates
Author:
Pool B.L., Lin P.Z.
Year:
1982
Bibliographic source:
Food and Chemical Toxicology, 20, 383-391.

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Guaiacol was purchased from Kluka, Buchs and was 98% pure or of analytical grade.

Method

Species / strain
Species / strain:
other: TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254 pre-treated Sprague-Dawley rats.
Test concentrations with justification for top dose:
0, 0.5, 5, 50, 500, 5000 µg/plate
Vehicle:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Negative controls:
no
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: see details below
Details on test system and conditions:
POSITIVE CONTROL(S) SUBSTANCE:
2-Nitrofluorene: 3 µg/plate for TA 98 and TA 1538 without metabolic activation
Sodium azide: 2 µg/plate for TA 1535 and TA 100 without metabolic activation
9-Aminoacridine: 60 µg/plate for TA 1537 without metabolic activation
2-Aminoanthracene: 5 µg/plate for all strains with metabolic activation

DETAILS ON TEST SYSTEM AND CONDITIONS:
S. Typhimurium strains TA 1535, TA 1537, TA 1538, TA 98, and TA 100 were kindly supplied by Prof. B.N. Ames (University of Berkeley, Calif., USA).
The test for their reversion to histidine prototrophy was performed using the plate incorporation assay as previously described (Ames, McCann & Yamasaki, 1975). Each compound was dissolved in DMSO and tested in 5 concentrations in the presence of buffer ph 7.4 or S9 mix.

METHOD OF APPLICATION: in agar (plate incorporation)
Exposure duration: 48 hours
Evaluation criteria:
The number of histidine revertants were scored and compared to the number of spontaneously arising revertants.

Results and discussion

Test results
Species / strain:
other: TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
the concentration of 5000 µg/plate resulted in toxicity which was apparent as a thinning of the background lawn.
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table of results:

Concentration [µg/plate]

No. of revertants per plate

No. of revertants per plate

Cytotoxicity

(yes/no)

 

— MA

+ MA

 

TA 100

0

67

76

no

0,5

66

94

no

5

71

93

no

50

65

81

no

500

67

95

no

5000

57

64

no

Sodium azide / 2-Aminoanthracene

502

723

no

TA 1535

0

24

9

no

0,5

21

16

no

5

26

13

no

50

22

19

no

500

15

16

no

5000

25

9

no

Sodium azide / 2-Aminoanthracene

526

306

no

TA 1537

0

5

14

no

0,5

7

12

no

5

7

16

no

50

4

13

no

500

4

9

no

5000

0

13

yes (-S9)

9-Aminoacridine / 2-Aminoanthracene

810

269

no

TA 98

0

19

19

no

0,5

20

34

no

5

18

36

no

50

23

14

no

500

22

16

no

5000

16

5

yes (+S9)

2-Nitrofluorene/ 2-Aminoanthracene

275

1093

no

TA 1538

0

5

10

no

0,5

13

12

no

5

9

11

no

50

10

11

no

500

14

9

no

5000

13

10

no

2-Nitrofluorene/ 2-Aminoanthracene

284

1027

no

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with and without metabolic activation

- INTERPRETATION OF RESULTS:
Regarding results and applying actual criteria evaluation, the test item presented no mutagenic activity with and without S9 mix in
Salmonella Typhimurium in all strains.
Reproductibility between the experiment and positive control results in accordance with positive criteria, validated the study.
Executive summary:

In a reverse gene mutation assay in bacteria (Pool et al., 1982), strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538 of S. typhimurium were exposed to guaiacol at concentrations of 0 to 5000 µg/plate in the presence and absence of mammalian metabolic activation.

Guaiacol was tested up to cytotoxic concentrations and was negative.

Cytotoxicity was observed at 5000 µg/plate.

The positive controls induced the appropriate responses in the corresponding strains. There was no evidence of induced mutant colonies over background.