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Genetic toxicity: in vitro

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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:
17 January 2005 - 24 January 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was performed in accordance with OECD, EC and Japanese test guidelines, and a certificate of GLP compliance is included in the report.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2005
Report date:
2005

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: "Commission Directive 2000/32/EC, L1362000, Annex 4D", dated May 19, 2000
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japanese Guidelines: "Kanpoan No. 287 - Environment Protection Agency" "Eisei No. 127 - Ministry of Health & Welfare" "Heisei 09/10/31 Kikyoku No. 2 - Ministry of International Trade & Industry"
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Issued by the German National GLP monitoring authority: Hessisches Ministerium für Umwelt, ländlichen Raum und Verbraucherschutz
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Polyol PX
IUPAC Name:
Polyol PX
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): Polyol PX
- Physical state: Yellowish liquid
- Analytical purity: >95% Mix of different polyols
- Composition of test material, percentage of components: Not specified
- Lot/batch No.: 2004-11-18
- Expiration date of the lot/batch: 18 November 2005
- Storage condition of test material: Room temperature

Method

Target gene:
Not applicable
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
- Type and identity of media:
- Properly maintained: yes/no
- Periodically checked for Mycoplasma contamination: yes/no
- Periodically checked for karyotype stability: yes/no
- Periodically "cleansed" against high spontaneous background: yes/no
Species / strain / cell type:
E. coli WP2 uvr A
Details on mammalian cell type (if applicable):
- Type and identity of media:
- Properly maintained: yes/no
- Periodically checked for Mycoplasma contamination: yes/no
- Periodically checked for karyotype stability: yes/no
- Periodically "cleansed" against high spontaneous background: yes/no
Metabolic activation:
with and without
Metabolic activation system:
S9 Mix
Test concentrations with justification for top dose:
Experiment I (Pre-experiment) - 3, 10, 33, 100, 333, 1000, 2500, and 5000 µg/plate
Experiment II - 33, 100, 333, 1000, 2500, and 5000 µg/plate.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: Solvent chosen based on the solubility of the test substance in this solvent.
Controlsopen allclose all
Untreated negative controls:
yes
Remarks:
Untreated control
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
For TA1535 and TA100 without metabolic activation.

Migrated to IUCLID6: 10 µg/plate
Untreated negative controls:
yes
Remarks:
Untreated control
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylene-diamine, 4-NOPD
Remarks:
For TA98 (10µg/plate) and TA1537 (50µg/plate) without metabolic activation
Untreated negative controls:
yes
Remarks:
Untreated control
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
For WP2 uvrA without metabolic activation.

Migrated to IUCLID6: 4µL/plate
Untreated negative controls:
yes
Remarks:
Untreated control
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene, 2-AA
Remarks:
TA strains 2.5µg/plate; WP2 uvrA 10µg/plate, with metabolic activation.
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)


DURATION
- Preincubation period: 60 minutes
- Exposure duration: 48 hours


NUMBER OF REPLICATIONS: 3 plates per dose level per strain.


DETERMINATION OF CYTOTOXICITY
- Method: other: reduction in the number of spontaneous revertants or a clearing of the bacterial background lawn.


OTHER EXAMINATIONS:
- Determination of polyploidy:
- Determination of endoreplication:
- Other:


OTHER:
Evaluation criteria:
Mutagenicity determined by examination of the number of revertants (revertant colonies).
Statistics:
No statistical evaluation of the data was required.

Results and discussion

Test resultsopen allclose all
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:
valid
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
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:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- None reported


RANGE-FINDING/SCREENING STUDIES:
A pre-experiment was performed in the range 3 - 5000 µg/plate. As no toxic effects were observed (no decrease in the number of spontaneous revertants) the results of the pre-experiment were reported as experiment I.

COMPARISON WITH HISTORICAL CONTROL DATA:
In experiment II, without metabolic activation, the data in the negative and solvent control of strain TA 100 were slightly above the lab's historical control range. Since this deviation is rather small, this effect is considered to be based upon biologically irrelevant fluctuations in the number of colonies.

ADDITIONAL INFORMATION ON CYTOTOXICITY:

No evidence of cytotoxicity was seen
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

None

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative With and without metabolic activation.

No evidence of mutagenicity was seen under the conditions of this study
Executive summary:

A bacterial reverse mutation assay was performed by RCC Cytotest Cell Research GmbH, Germany, on behalf of Perstorp Specialty Chemicals AB, Sweden, to determine the potential of the test substance Polyol PX to induce gene mutations. The study was performed according to the relevant OECD 471 Test Guideline, and in accordance with GLP. Four strains of Salmonella typhimurium and one strain of Escherichia coli were tested, both in the presence and absence of metabolic (S9) activation. None of the strains tested at any of the test concentrations showed any signs of cytotoxicity, or signs of increased gene mutation. All appropriate reference mutagens, used as positive controls, produced increased numbers of revertant colonies. Under the test conditions applied, the test substance Polyol PX did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used. Therefore, Polyol PX is considered to be non-mutagenic in this Salmonella typhimurium and Escherichia coli reverse mutation assay.