Reaction mass of 2-ethylpropane-1,3-diol and 5-ethyl-1,3-dioxane-5-methanol and propylidynetrimethanol

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:

08 May 2008 - 13 June 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed according to OECD, EC, US EPA, Japanese and other international test guidelines, and a certificate of GLP compliance has been included.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries. Test Data for Registration of Agricultural Chemicals, 12 Noshan No. 8417, Guideline 2-1-19-1, Agricultural Production Bureau, November 24, 2000

Deviations:

no

Test guideline 5

Qualifier:

according to guideline

Guideline:

other: Official notice of J MHLW, METI and ME (21 November 2003)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Includes MHRA GLP compliance certificate.

Type of assay:

bacterial reverse mutation assay

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): Polyol TD
- Substance type:
- Physical state: Colourless liquid
- Analytical purity: Hydroxyl number, 760 mg KOH/g
- Impurities (identity and concentrations): Not reported.
- Composition of test material, percentage of components: Not reported.
- Isomers composition: Not reported.
- Purity test date: Not reported.
- Lot/batch No.: 3777937
- Expiration date of the lot/batch: Indefinite
- Stability under test conditions: Not reported.
- Storage condition of test material: Room temperature in the dark, dry
- Other:

Method

Target gene:

N/A

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

5, 15, 50, 150, 500, 1500, 5000 µg/plate, first test.
50, 150, 500, 1500, 5000 µg/plate, second test.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water, purified in-house by reverse osmosis
- Justification for choice of solvent/vehicle: The Sponsor indicated that the test substance was miscible with water.

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

: water

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

In the absence of S9 mix

Migrated to IUCLID6: 2 µg/plate for strains TA100 and TA 1535

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

: water

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

In the absence of S9 mix

Migrated to IUCLID6: 50 µg/plate for strain TA1537

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

: water

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

In the absence of S9 mix

Migrated to IUCLID6: 2 µg/plate for strain TA98

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

: water

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

In the absence of S9 mix

Migrated to IUCLID6: 2 µg/plate for strain WP2 uvrA (pKM101)

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

: water

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene, 5 µg/plate for strains TA 100 and TA 1535; 10 µg/plate for strain WP2 uvrA (pKM101)

Remarks:

In the presence of S9 mix

Controls 6

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

: water

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

In the presence of S9 mix

Migrated to IUCLID6: 5 µg/plate for strains TA98 and TA 1537

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation) - first test; preincubation - second test


DURATION
- Preincubation period: 30 minutes at 37ºC (second test only).
- Exposure duration: Plates incubated for 72 hours
- Expression time (cells in growth medium): 10 hours
- Selection time (if incubation with a selection agent): N/A
- Fixation time (start of exposure up to fixation or harvest of cells): N/A


SELECTION AGENT (mutation assays): Histidine (Salmonella strains) and tryptophan (E Coli strain)
SPINDLE INHIBITOR (cytogenetic assays): N/A
STAIN (for cytogenetic assays): N/A


NUMBER OF REPLICATIONS: Three plates at each concentration in each test.


NUMBER OF CELLS EVALUATED: N/A


DETERMINATION OF CYTOTOXICITY
- Method: Any toxic effects of the test substance would be detected by a substantial reduction in mean revertant colony counts or by a sparse or
absent backgroud bacterial lawn.


OTHER EXAMINATIONS:
- Determination of polyploidy: N/A
- Determination of endoreplication: N/A
- Other:


OTHER:

Evaluation criteria:

If exposure to a test substance produces a reproducible increase in revertant colony numbers of at least twice (three times in the case of strains TA1535 and TA1537) the concurrent vehicle controls, with some evidence of a positive dose-response relationship, it is considered to exhibit mutagenic
activity in this test system. No statistical analysis is performed.
If exposure to a test substance does not produce a reproducible increase in revertant colony numbers, it is considered to show no evidence of mutagenic activity in this test system. No statistical analysis is performed.

Statistics:

If the results obtained fail to satisfy the criteria for a clear “positive” or “negative” response, even after additional testing, the test data may be subjected to analysis to determine the statistical significance of any increases in revertant colony numbers. The statistical procedures used are those described by Mahon et al (1989) and are usually Dunnett’s test followed, if appropriate, by trend analysis. Biological importance should always be considered along with statistical significance. In general, treatment-associated increases in revertant colony numbers below two or three times the vehicle controls (as described above) are not considered biologically important. It should be noted that it is acceptable to conclude an equivocal response if no clear results can be obtained.

Results and discussion

Test results

Test results 1

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: Not reported.
- Effects of osmolality: Not reported.
- Evaporation from medium: Not reported.
- Water solubility: Not reported.
- Precipitation:Not reported.
- Other confounding effects: Not reported.


RANGE-FINDING/SCREENING STUDIES:
No evidence of toxicity was obtained following exposure to Polyol TD. A maximum exposure concentration of 5000 μg/plate was, therefore, selected for use in the second test.

COMPARISON WITH HISTORICAL CONTROL DATA:
The mean revertant colony counts for the vehicle controls were within or close to the 99% confidence limits of the current historical control range of the laboratory.

ADDITIONAL INFORMATION ON CYTOTOXICITY:

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Refer to results tables, attached.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

It is concluded that Polyol TD showed no evidence of mutagenic activity in this bacterial system under the test conditions employed.

Executive summary:

A bacterial reverse mutation test was performed by Huntingdon Life Sciences, UK, on behalf of Perstorp Specialty Chemicals AB, Sweden, to determine the potential of the test substance Polyol TD to induce gene mutation. The test was performed in accordance with OECD, EC, US EPA, Japanese, and other international test guidelines, and the test was carried out to GLP. No evidence of cytotoxicity or mutagenic activity was observed during two tests, either in the presence or absence of metabolic activation at concentrations up to and including the limit concentration of 5000 ug/plate.