DL-hexane-1,2-diol

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:

15-Jun, 1998 to 6-August, 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: guideline test with GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

DL-hexane-1,2-diol

EC Number:

230-029-6

EC Name:

DL-hexane-1,2-diol

Cas Number:

6920-22-5

Molecular formula:

C6H14O2

IUPAC Name:

hexane-1,2-diol

Constituent 2

Reference substance name:

1,2-hexanediol

IUPAC Name:

1,2-hexanediol

Test material form:

other: liquid

Method

Target gene:

S. typhimurium histidine (his) reversion system measures his- → his+ reversions.
E. coli WP2 and E.coli wp2 uvr A pKM101

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Additional strain / cell type characteristics:

not specified

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Additional strain / cell type characteristics:

not specified

Species / strain 3

Species / strain / cell type:

E. coli WP2 uvr A pKM 101

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver s9

Test concentrations with justification for top dose:

the preliminary toxicity assay: 6.7, 10, 33, 67, 100,333, 667, 1000, 3333, 5000 µg/plate;
the mutagenicity assays: 25, 75, 200, 600, 1800, 5000 µg/plate

Vehicle / solvent:

water was selected as the solvent of choice based on solubility of the test article and compatibility with the target cell.

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

with s9

Positive control substance:

other: 2-aminoanthracene

Remarks:

1.0 µg/plate for all salmonella strains

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

with s9

Positive control substance:

other: 2-aminoanthracene

Remarks:

10 µg/plate for WP2 uvrA (pKM101)

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

with s9

Positive control substance:

other: sterigmatocystin

Remarks:

100 µg/plate for WP2(pKM)101

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

without s9

Positive control substance:

2-nitrofluorene

Remarks:

1.0 µg/plate for TA98, TA 1538

Controls 5

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

without s9

Positive control substance:

sodium azide

Remarks:

1.0 µg/plate for TA100, TA1535

Controls 6

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

without s9

Positive control substance:

other: 9-aminoacridine

Remarks:

75 µg/plate for TA1537

Controls 7

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

without s9

Positive control substance:

methylmethanesulfonate

Remarks:

1000 µg/plate for Both E.coli strains

Details on test system and experimental conditions:

Test system:
The tester strains used were the salmonella typhimurium histidine auxotrophs TA 98, TA 100, TA 100, TA 1537 and TA 1538 as described by Ames etal.(1975) and Escherichia cloi tester strains WP2 uvrA (PKM 101) and WP2(PKM 101). Salmonella tester strains were received on 11/10/92 directly from Dr. Bruce Ames, University of California, Berkeley. E.coli was received on 07/01/87 and 02/19/93 from the National Collection of Industrial and Marine Bacteria, Aberdeen, Scotland.
Tester strains TA 98, TA 1537 and TA 1538 are reverted from histidine dependence(auxotrophy) to histidine independence(prototrophy) by frameshift mutagens. Tester strain TA 1535 is reverted by mutagens that cause basepair substitutions. Tester strain TA100 is reverted by mutagens that cause both frameshift and basepair substitution mutations. Specificity of the reversion mechanism in E.coli is sensitive to base-pair substitution mutations, rather than frameshift mutations.
Metabolic activation system
Aroclor 1254-induced rat liver s9 was used as the metabolic activation system. The s9 was prepared from male sprague-Dawley rats induced with a single intraperitoneal injection of Aroclor 1254, 500mg/kg, five days prior to sacrifice. The s9 was batch prepared on 02/25/98 and 02/26/98 and stored at <=-70 degree until used. Each bulk preparation of s9 was assayed for its ability to metabolized 2-aminoanthracene and 7,12-dimethylbenz(a)anthracene to forms mutagenic to Salmonella typhimurium TA 100.

Evaluation criteria:

For the test article to be evaluated positive, it must cause a dose-related increase in the mean revertants per plate of at least one tester strain with a minimum of two increasing concentrations of test article. Data sets ofr strains TA1535, TA1537 and TA1538 were judged positive in the increase in mean revertants at the peak of the dose response is equal to or greater than three times the mean vehicle control value. Data sets for strains TA98, TA100 and WP2 uvrA(pKM101) and WP2 (pKM101) were judged positive if the increase in mean revertants at the peak of the dose response is equal or greater than two times the mean vehicle control value.

Statistics:

Revertant colonies for a given tester strain and activation condition, except for positive controls, were counted either entirely by automated colony counter or entirely by hand. For each replicate plating, the mean and standard deviation of the number of revertants per plate were calculated and are reported.

Results and discussion

Test results

Test results 1

Species / strain:

other: TA 98, TA100, TA1535, TA1537 and TA1538

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

toxicity was observed at the 5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

other: TA 98, TA100, TA1535, TA1537 and TA1538

Metabolic activation:

with

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:

other: E. coli WP2 uvr A pKM 101 and E. coli WP2 pKM 101

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

Additional information on results:

slubility test:
This test was conducted with the vehicle. The test article was soluble in water at approximately 500 mg/ml, the maximum concentration tested.
Priliminary toxicty assay
In the preliminary toxicity assay, the maximum dose tested was 5000 µg/plate; this dose was achieved using a concentration of 100 mg/ml and a 50 µl plating aliquot. No precipitate was observed but toxicity was generally observed at 5000 µg/plate with tester strains TA 98, TA100, TA1535, TA1537 and TA1538 in the absence of s9 only. Based on the findings of the toxicity assay, the maximum dosed plated in the mutagenicity assay was 5000 µg/plate.
In Experiment B1, the initial mutagenicity assay, no positive responses were observed with any of the tester strains in the presence of s9 and with tester strains TA1538,WP2uvrA(pKM101) and WP2pKM 101 in the absence of s9.
In experiment B2, no positive responses were observed with tester strains TA1535 and TA1537 without s9.
In experiment B3,no positive responses were observed with tester strain TA98 without s9.
In experiment B4, no positive responses were observed with tester strains TA98, TA100, TA1535, TA1537 and WP2 (pKM101) with s9. and with TA98, TA100, TA1535, TA1537 without s9.
In experiment B5, no positive responses were observed with tester strains TA1538, WP2uvrA(pKM101) and WP2pKM 101 with and without s9.

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 and without metabolic activation

The results of the Salmonella/Mammalian Microsome (Ames test) and Escherichia coli WP2 mutagenesis assay indicate that test article did not cause a positive respone with any of the tester strains with and without metabolic activation under the conditions of this study.

Executive summary:

This study was conducted following OECD guideline 471 with GLP regulations. Test article was tested in the bacterial reverse mutation assay using S.typhimurium tester strains TA98, TA100, TA 1535, TA 1537 TA1538 E.coli tester strains WP2 uvrA (pKM101) and WP2 (pKM101) in the presence and absence of Aroclor-induced rat liver S9. The assay was performed in two phases, using the plate incorporation and preincubation methods.In the preliminary toxicity assay, test article was tested at levels of 6.7, 10, 33, 67, 100, 333, 667, 1000, 3333, 5000 µg/plate. Based on the results of preliminary test, the mutagenicity assays was conducted at dose levels of 25, 75, 200, 600, 1800, 5000 µg/plate. All dose levels of test article and controls were plated in triplicate. All criteria for a valid study were met.

Based on the results of mutagenesis assay, test article caused negative response with any of the tester strains in the absence and presence of metabolic activation under the conditions of this study.