2-(2H-benzotriazol-2-yl)-4-(1,1,3,3-tetramethylbutyl)phenol

Genetic toxicity: in vitro

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: Guideline study (non-GLP), no confirmatory experiment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

(no confirmatory experiment)

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-(2H-benzotriazol-2-yl)-4-(1,1,3,3-tetramethylbutyl)phenol

EC Number:

221-573-5

EC Name:

2-(2H-benzotriazol-2-yl)-4-(1,1,3,3-tetramethylbutyl)phenol

Cas Number:

3147-75-9

Molecular formula:

C20H25N3O

IUPAC Name:

2-(2H-1,2,3-benzotriazol-2-yl)-4-(2,4,4-trimethylpentan-2-yl)phenol

Test material form:

solid

Details on test material:

- Name of test substance (as cited in study report): Tinuvin 329
- Analytical purity: >99%
- Batch No.: EN 171744.92
- Physical state: solid
- Storage conditions: room temperature
- Validity: November 1993

Specific details on test material used for the study:

- Name of test substance (as cited in study report): Tinuvin 329
- Analytical purity: >99%
- Batch No.: EN 171744.92
- Stability: checkt via reanalysis

Method

Target gene:

his- and trp-operon

Species / strain

Species / strain / cell type:

other: Salmonella typhimurium: TA 98, TA 100, TA 1535, TA 1537; Escherichia coli: WP2uvrA

Metabolic activation:

with and without

Metabolic activation system:

S9-mix: Aroclor-induced rat liver S9 fraction (1 part) and co-factors: NADP, MgCl2, KCl, Glucose-6-phosphate in Na-phosphate buffer) (9 parts)

Test concentrations with justification for top dose:

Concentration range in the cytotoxicity/range finding test: 20.6 - 5,000 µg/plate
Concentration range in the mutagenicity test: 625 - 5,000 µg/plate (with and without metabolic activation)

Vehicle / solvent:

DMSO

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: (- S9-mix): sodium azide (TA 100 and TA 1535), 4-nitroquinoline-N-oxide (WP2uvrA), 2-nitrofluorene (TA 98), 9(5)-aminoacridine (TA1537); (+S9-mix): 2-aminoanthracene (TA 100, TA 98, TA 1537, WP2uvrA), cyclophosphamide (TA 1535)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation)

DURATION
- Exposure duration: 48 h at 37 °C

NUMBER OF REPLICATIONS: 2 plates/dose

DETERMINATION OF CYTOTOXICITY
- Method: number of colonies and determination of background lawn in comparison to the corresponding control values

OTHER: range finding test
- Strains: S. typhimurium TA 100 and E. coli WP2uvrA
- Concentrations: 20.6, 61.7, 185.2, 555.6, 1666.7 and 5000.0 µg/plate
- Number of replications: 1 plate/dose
- Exposure duration: 48 h
5000 µg/plate was chosen as highest dose for the mutagenicity assay.

Evaluation criteria:

Criteria for a positive response: The test substance is considered to be mutagenic in this test system if:
- A positive effect is observed in one strain and the effect can be reproduced in a confirmatory experiment.
- A positive effect is observed in two or more strains.
A positive effect is defined as an increase of the mean number of revertants per plate for any concentration above that of the negative control by at least a factor of 2.0 with strains TA 98, TA 1535, TA 1537 and WP2uvrA, or by a factor of at least 1.5 with strain TA 100. Generally a concentration-related effect should be demonstrable.

Acceptance critaria: A test is considered acceptable if the mean colony counts of the control values of all strains are within the acceptable ranges and if the results of the positve controls meet the criteria for a positive resoponse.

Results and discussion

Test results

Species / strain:

other: Salmonella typhimurium: TA 98, TA 100, TA 1535, TA 1537; Escherichia coli: WP2uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

(reduced number of revertants at 5000 µg/plate in TA 98 without metabolic activation: factor 0.39 compared to control)

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

- Precipitation: Precipitation occurred at concentrations >= 1666.7 µg/plate

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Standard plate test:

Dose (µg/plate)	TA100		TA1535		TA1537		TA98		E. coli WP2 uvrA
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
0	166.5	140.0	10.0	12.5	6.5	12.5	20.5	40.5	19.5	22.0
625	161.5	141.5	8.5	13.0	8.0	12.0	15.5	31.5	13.0	23.5
1250	147.0	123.0	11.0	13.0	6.5	13.0	18.5	27.5	13.5	15.0
2500	143.5	120.0	6.5	9.0	7.0	11.5	18.0	25.5	14.5	13.0
5000	128.5	113.0	6.5	9.0	6.0	7.5	8.0	30.0	18.0	15.5
Sodium azide	1778.0	-	1452.5	-	-	-	-	-	-	-
4-NQO	-	-	-	-	-	-	-	-	847.0	-
2-nitrofluorene	-	-	-	-	-	-	1278.5	-	-	-
AAC	-		-	-	1491.5	-	-	-	-	-
2-AA	-	1936.0	-	-	-	249.0	-	2239.5	-	1015.0
Cyclophosphamide	-	-	-	377.0	-	-	-	-	-	-

Mean ± SD 

X: reduced background growth

4-NQO: 4-nitroquinoline-N-oxide

AAC: 9-aminoacridine chloride monohydrate

2-AA: 2-aminoanthracene

 

 

According to the results of the study, the test substance is not mutagenic in the Ames test and in the Escherichia coli - reverse mutation assay under the experimental conditions chosen here. The positive controls gave the expected values.

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative