Butane-1,3-diol

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:

not stated

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Reference 2

Reference Type:

publication

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

not stated

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

Version / remarks:

not stated

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: “Methods of Testing New Chemical Substances” (Environmental Protection Bureau notification no. 237, Pharmaceutical Affairs Bureau notification no. 306, Basic Industries Bureau 1987 notification no. 303, dated 31 March, 1987

Version / remarks:

31 March 1987

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Butane-1,4-diol

EC Number:

203-786-5

EC Name:

Butane-1,4-diol

Cas Number:

110-63-4

Molecular formula:

C4H10O2

IUPAC Name:

butane-1,4-diol

Test material form:

liquid

Specific details on test material used for the study:

Purity of test material: 99.8 wt% (impurities: 0.06 wt% 1,4-acetoxyhydroxybutane-2, 0.07 wt% 2-(4-hydroxy-butyloxy)tetrahydrofuran)

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Additional strain / cell type characteristics:

not specified

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 (Kikkoman Co., Ltd., lot no: RAA-333, manufactured 8 September, 1995 and lot no: RAA-338, 15 December, 1995) extracted from oxygen-induced 7-week old Sprague-Dawley male rats administered phenobarbital (PB) and 5,6-benzoflavone (BF

Test concentrations with justification for top dose:

313~5000 μg/plate with a common ratio of 2, top dose according to guideline

Vehicle / solvent:

water for injection use

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: AF2: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide; SA: Sodium azide; 9AA: 9-aminoacridine; 2AA: 2-aminoanthracene

Details on test system and experimental conditions:

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

DURATION
- Preincubation period: 20 min at 37 °C
- Exposure duration: 48 h
- Fixation time (start of exposure up to fixation or harvest of cells): 48 h

NUMBER OF REPLICATIONS:main test and repeatability test, 3 plates were used for both control groups and for each dose

DETERMINATION OF CYTOTOXICITY
not stated

Evaluation criteria:

If, for 1 or more of the 5 strains of bacteria used, under conditions without S9 mix and with S9 mix, the mean value of the revertant colony count on the surface of plates containing the test substance was 2 or more times that of the solvent control, and, if that increase could be repeated or dose-dependency was noted, then the test substance in this series of tests would be determined to be mutagenic (positive). However, if the dose showed that the mean value of the revertant colony count at the second time of testing was more than 2 times that of the solvent control, but the solvent control value was less than 10 and a dose-dependent increase in the revertant colony count was not noted, then the determination would be negative.

Statistics:

not performed

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Remarks:

negative in range finding test with same top dose

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Remarks:

negative in range finding test with same top dose

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Remarks:

negative in range finding test with same top dose

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Remarks:

negative in range finding test with same top dose

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Remarks:

negative in range finding test with same top dose

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other:

Remarks:

Without the addition of S9 mix, the result in Test I for the 625 μg/plate showed an increase in revertant colony count which was more than twice that of the solvent control value. However, in the testing of TA1537 with the addition of S9 mix and in the tests with all the other bacteria, there was no increase in revertant colony count more than twice that of the solvent control. The main test was repeated using identical doses. As a result, no increase in revertant colony count more than twice that of the solvent control was noted.

Applicant's summary and conclusion

Conclusions:

Under the conditions of this study the test substance was not mutagenic to bacteria with and without metabolic activation.

Executive summary:

A bacterial mutagenicity test (according to OECD guidelines 471 and 472 and GLP-compliant) was performed in Salmonella typhimurium TA1535, TA1537, TA98 and TA100, and in Escherichia coli WP2 uvrA as preincubation test, with and without metabolic activation.

Tests with and without S9 mix were conducted twice, within the range of 313 -5000 μg/plate with a common ratio of 2.

For the test of TA1537, without the addition of S9 mix, the result in Test I for the 625 μg/plate showed an increase in revertant colony count which was more than twice that of the solvent control value. However, in the testing of TA1537 with the addition of S9 mix and in the tests with all the other bacteria, there was no increase in revertant colony count more than twice that of the solvent control.

For TA1537, since the results without S9 mix were different for Test I and Test II, the main test was repeated using identical doses. As a result, no increase in revertant colony count more than twice that of the solvent control was noted.

In all of the tests conducted, the positive control group showed an increase in revertant colony count for all of the test bacteria, which meant that, along with the values measured for the solvent control group, the revertant colony count was within the range of historical values, so the efficacy of this test series was confirmed.

It is concluded, based on the above results that the test material is not mutagenic (negative) for the test strains used.