1,3-Benzenediol, 4-butyl-

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

Study period:

2010-03-04 to 2010-05-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study completed according to OECD guideline 471 (Bacterial Reverse Mutation Assay).

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Method

Target gene:

Histidine locus

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA1535, TA97A, TA98, TA100 and TA102

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

other: see below

Metabolic activation:

with and without

Metabolic activation system:

Phenobarbital/B-naphthoflavone induced rat liver S9

Test concentrations with justification for top dose:

5, 15, 50, 150, 500 ug/plate

Vehicle / solvent:

Vehicle used : dimethyl sulfoxide

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Without metabolic activation

Migrated to IUCLID6: Used for strain TA1535 (2ug/plate)

Controls 2

Positive controls:

yes

Positive control substance:

other: ICR. Used for strain TA97A (1ug/plate)

Remarks:

Without metabolic activation

Controls 3

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

Without metabolic activation

Migrated to IUCLID6: Used for strain TA98 (0.5ug/plate)

Controls 4

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

Without metabolic activation

Migrated to IUCLID6: Used for strains TA100 and TA102 (1ug/plate)

Controls 5

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene. Used for strain TA1535 (10ug/plate)

Remarks:

With metabolic activation

Controls 6

Positive controls:

yes

Positive control substance:

other: 2-Aminofluorene. Used for strains TA97A, TA98 and TA100 (20ug/plate)

Remarks:

With metabolic activation

Controls 7

Positive controls:

yes

Positive control substance:

other: Danthron. Used for strain TA102

Remarks:

With metabolic activation

Details on test system and experimental conditions:

The plate incorporation method was followed for the test. Suspensions of bacterial cells were exposed to the test article in the presence and in the absence off an exogenous metabolic activation system. These suspensions were mixed with an overlay agar and plated immediately onto minimal medium. After 48 to 72 hours incubation period revertant colonies were counted and compared to the number of spontaneous revertant colonies on vehicle control plates. In order to confirm the reproducibility of the results the entire study was carried out twice as experiment number 1 and experiment number 2.

Evaluation criteria:

A test article is considered as mutagenic if it produces a concentration related increase over the range tested and/or a reproducible increase at one or more concentrations in the number of revertant colonies per plate in at least one strain with or without metabolic activation system. The biologically significant increase would be assumed if the average colony counts are at least twice as compared to those in vehicle control group for strain TA97A, TA98, TA100 and TA102 and at least thrice as compared to those in vehicle control group for strain TA1535. A test article is considered as non-mutagenic, if it does not induce any increase in the number of revertants and does not show any dose response relationship, in two separate experiments with any bacterial strain, either with or without metabolic activation.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA1535, TA97A, TA98, TA100 and TA102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

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

Salmonella typhimurium, Reverse Mutation Assay of 4-butylbenzene-1,3-diol was carried out in compliane with the OECD Guidelines for Testing of Chemicals (No 471, section 4, Health Effects) on conduct of Bacterial Reverse Mutation Test, adopted 21 July 1997, OPPTS Guideline 870.5100 and as per mutually agreed protocol. Vivinol was evaluated in the Ames/Salmonella Plate Incorporation Assay to determine its ability to induce reverse mutation at selected histidine loci in five tester strains of Salmonella typhimurium TA1535, TA97A, TA98, TA100 and TA102 in the presence and absence of metabolic activation system (S9). Based upon the preliminary tests conducted to assess the solubility/precipitation and cytotoxicity of the test substance, the tester strains were exposed to the test article in triplicate cultures at the doses of 500ug, 150ug, 50ug, 15ug and 5ug/plate, both with and without metabolic activation system (S9). Liver S9, induced in Sprague Dawley rats by phenobarbitone with B-naphthoflavone, was used for this purpose. Dimethyl sulfoxide was used as a vehicle. The exposed bacteria were plated onto minimal glucose agar medium supplemented with L-histidine. The plates were incubated at 37 deg C for 48-72 hours after which the histidine revertant colonies were counted and their frequency was compared with that in the vehicle control group. Concurrent negative control group and positive control groups were also included in the experiment, as specified by the test guideline. Results of this test indicated that the frequencies of histidine revertant colonies at all concentrations of 4-butylbenzene-1,3-diol in strains TA1535, TA97A, TA98, TA100 and TA102, with and without the presence of metabolic activation system, were comparable to those observed in the vehicle control group, as per criteria employed for evaluation of mutagenic potential, and this observation was confirmed by repetition of the experiments. Plate counts for the spontaneous histidine revertant colonies in the vehicle control groups were found to be within the frequency ranges expected from the laboratory historical control data at the test facility. They also compared well with the range reported in the literature. Concurrent positive controls demonstrated sensitivity of the assay with and without metabolic activation. It is concluded that, under the conditions of the study, 4-butylbenzene-1,3-diol is non-mutagenic in Salmonella typhimurium strains TA1535, TA97A, TA98, TA100 and TA102.

Executive summary:

Salmonella typhimurium, Reverse Mutation Assay of 4 -butylbenzene-1,3 -diol was carried out in compliane with the OECD Guidelines for Testing of Chemicals (No 471, section 4, Health Effects) on conduct of Bacterial Reverse Mutation Test, adopted 21 July 1997, OPPTS Guideline 870.5100 and as per mutually agreed protocol. 4 -Butylbenzene-1,3 -diol was evaluated in the Ames/Salmonella Plate Incorporation Assay to determine its ability to induce reverse mutation at selected histidine loci in five tester strains of Salmonella typhimurium TA1535, TA97A, TA98, TA100 and TA102 in the presence and absence of metabolic activation system (S9). Based upon the preliminary tests conducted to assess the solubility/precipitation and cytotoxicity of the test substance, the tester strains were exposed to the test article in triplicate cultures at the doses of 500ug, 150ug, 50ug, 15ug and 5ug/plate, both with and without metabolic activation system (S9). Liver S9, induced in Sprague Dawley rats by phenobarbitone with B-naphthoflavone, was used for this purpose. Dimethyl sulfoxide was used as a vehicle. The exposed bacteria were plated onto minimal glucose agar medium supplemented with L-histidine. The plates were incubated at 37 deg C for 48-72 hours after which the histidine revertant colonies were counted and their frequency was compared with that in the vehicle control group. Concurrent negative control group and positive control groups were also included in the experiment, as specified by the test guideline. Results of this testindicated that the frequencies of histidine revertant colonies at all concentrations of 4 -butylbenzene-1,3 -diol in strains TA1535, TA97A, TA98, TA100 and TA102, with and without the presence of metabolic activation system, were comparable to those observed in the vehicle control group, as per criteria employed for evaluation of mutagenic potential, and this observation was confirmed by repetition of the experiments. Plate counts for the spontaneous histidine revertant colonies in the vehicle control groups were found to be within the frequency ranges expected from the laboratory historical control data at the test facility. They also compared well with the range reported in the literature. Concurrent positive controls demonstrated sensitivity of the assay with and without metabolic activation. It is concluded that, under the conditions of the study, 4 -butylbenzene-1,3 -diol is non-mutagenic in Salmonella typhimurium strains TA1535, TA97A, TA98, TA100 and TA102.