Vanillin

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 16 dec 1990 to 09 may 1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study is sufficiently documented and the test conditions are similar to current test guidelines. The study was conducted according to GLP. The purity of the test substance is unknown.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Vanillin

EC Number:

204-465-2

EC Name:

Vanillin

Cas Number:

121-33-5

Molecular formula:

C8H8O3

IUPAC Name:

1-butoxypropan-2-ol

Method

Species / strain

Species / strain / cell type:

other: Salmonella typhimurium TA98, TA100, TA1535, TA1537, TA1538

Metabolic activation:

with and without

Metabolic activation system:

S9 from induced Aroclor 1254

Test concentrations with justification for top dose:

100, 333, 667, 1000, 3330, and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: no data

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 2-nitrofluorene (1 µg/pl): in TA 98 and TA 1538 without S9 mix; sodium azide (2 µg/pl): in TA 100 and TA 1535 without S9 mix; ICR 191 (2 µg/pl): in TA 1537 without S9 mix; 2-aminoanthracene (2.5 µg/pl): in all strains with S9 mix

Details on test system and experimental conditions:

- Method of application: in agar (plate incorporation)
- Duration:
* Preincubation period: none
* Exposure duration: 48 +/- 8 hrs
- Number of replicates: 3

Evaluation criteria:

- in TA 98 and TA 100: a test article is considered as positive if it produce at least a 2-fold increase in the mean revertant per plate of at least one of
these tester strains over the mean revertants per plate of the appropriate vehicle control. This increase in the number of revertant must be
accompanied by a dose response with increasing concentrations of the test article
- in TA 1535, TA 1537 and TA 1538: idem, but a 3-fold increase is required

Statistics:

no data

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

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:

not applicable

Positive controls validity:

valid

Test results 4

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:

not applicable

Positive controls validity:

valid

Test results 5

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:

not applicable

Positive controls validity:

valid

Any other information on results incl. tables

	TA 98			TA 100			TA 1535
Conc. [unit]	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)
0*	14	22	no	94	120	no	17	14	no
100	14	22	no	97	115	no	13	14	no
333	9	27	no	92	111	no	12	6	no
667	10	26	no	79	106	no	10	12	no
1000	12	17	no	85	97	no	15	10	no
3330	13	17	no	65	80	no	10	13	no
5000	12	19	no	77	82	no	10	13	no
Positive control	209	998	no	602	1020	no	542	100		no

	TA 1537			TA 1538
Conc. [unit]	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)
0*	4	6	no	13	17	no
100	7	9	no	18	13	no
333	4	9	no	11	16	no
667	8	7	no	12	16	no
1000	4	6	no	13	17	no
3330	7	5	no	13	13	no
5000	5	6	no	9	15	no
Positive control	194	118	no	243	1112	no

*solvent control with DMSO

Applicant's summary and conclusion

Conclusions:

Negative with and without metabolic activation.

Executive summary:

In a reverse gene mutation assay in bacteria (Lawlor, 1991), strains TA98, TA 100, TA 1535, TA 1537 and TA 1538 of S. typhimurium were exposed to Vanillin (purity unknown), in DMSO at concentrations of 100, 333, 667, 1000, 3330 and 5000 µg/plate in the presence and absence of mammalian metabolic activation (plate co-incubation method). Vanillin was tested up to the limit concentration (5000 µg/plate).

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

This study is classified as acceptable. This study satisfies the requirement for Test Guideline EU B.13/14 for in vitro mutagenicity (bacterial reverse gene mutation) data.