Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
adopted July 21, 1997
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
dated May 30, 2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Version / remarks:
August, 1998
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
D-Glucose, reaction products with alcohols C16-18 (even numbered)
IUPAC Name:
D-Glucose, reaction products with alcohols C16-18 (even numbered)
Test material form:
solid: particulate/powder

Method

Target gene:
histidine operon
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
S9-liver mix
Test concentrations with justification for top dose:
10.0, 31.6, 100, 316, 1000, 2500 and 5000 pg/plate, the max. concentration was chosen due to results from a pre-experiment.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The solvent was compatible with the survival of the bacteria and the S9 activity.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
methylmethanesulfonate
other: 4-NOPD; 4-nitro-o-phenylene-diamine; 4-NOPD; without metabolic activation, 2-AA; 2-aminoanthracene; with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); preincubation
- Cell density at seeding (if applicable): approx. 1E09 cells/mL

DURATION
- Preincubation period: 60 min
- Exposure duration: 48 h in the dark

NUMBER OF REPLICATIONS: three


DETERMINATION OF CYTOTOXICITY
- Method: other: Cytotoxicity can be detected by a clearing or rather diminution of the background lawn or a reduction in the number of revertants down to a mutation factor of approximately < 0.5 in relation to the solvent control.

Rationale for test conditions:
The OECD Guideline for Testing of Chemicals, Section 4, No. 471 - Bacterial Reverse Mutation Test - recommends using a combination of S. typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 102.
Evaluation criteria:
A test item is considered as mutagenic if
- a clear and dose-related increase in the number of revertants occurs and/or
- a biologically relevant positive response for at least one of the dose groups occurs in at least one tester strain with or without metabolic activation.
A biologically relevant increase is described as follows:
- if in tester strains TA 98, TA 100 and TA 102 the number of reversions is at least twice as high
- if in tester strains TA 1535 and TA 1537 the number of reversions is at least three times higher than the reversion rate of the solvent control.

Results and discussion

Test results
Key result
Species / strain:
S. typhimurium, other: TA 98, TA 100, TA 102, TA 1535, TA 1537
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
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no data
- Effects of osmolality: no data
- Evaporation from medium: no data
- Precipitation: not described
- Other confounding effects: no

RANGE-FINDING/SCREENING STUDIES: yes, a range finding study was performed in the strains TA 98 and TA 100.

HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%)
- Positive historical control data: yes, the positive control plates were within the historical data range.
- Negative (solvent/vehicle) historical control data: yes, the negative control plates were within the historical data range.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
Cytotoxicity was observed in TA 100, TA 1535 and TA 1537 strains at a concentration of 5000 µg/plate, regardless whether the substance was tested with or without metabolic activation. However, there were no relevant increases in revertant colony numbers of any of the five tester strains following treatment with the test item at any concentration level, neither in the presence nor absence of metabolic activation.

Applicant's summary and conclusion

Conclusions:
D-Glucose, reaction products with alcohols C16-18 was evaluated in the bacterial reverse mutation assay (Ames test) using Salmonella typhimurium tester strains TA98, TA100, TA102, TA1535 and TA 1537 in the presence and absence of rat liver S9 mix. Under the conditions of the study, the test substance was negative for mutagenic potential.
Executive summary:

In a reverse gene mutation assay in bacteria according to OECD guideline 471 (adopted 21 July 1997), Salmonella typhimurium strains TA98, TA100, TA102, TA1535 and TA 1537 were exposed either to D-Glucose, reaction products with alcohols C16-18 in DMSO in concentrations of 0 (control), 31.6, 100, 316, 1000, 2500 and 5000 µg/plate in all strains in the absence and presence of mammalian metabolic activation (rat liver S9 mix) and in concentrations of 0 (control), 10.0, 31.6, 100, 316, 1000, 2500 and 5000 µg/plate in all strains in the absence and presence of mammalian metabolic activation (rat liver S9 mix). The assay was performed using the plate incorporation method and the preincubation method.

The test substance was tested up to cytotoxic concentrations. Cytotoxicity was observed in TA 100, TA 1535 and TA 1537 strains at a concentration of 5000 µg/plate. Precipitation was not observed.The positive controls induced the appropriate responses in the corresponding strains. The mean numbers of revertant colonies in the negative controls were within the ranges of the historical control data.

There was no evidence of an increase in the number of revertant colonies that exceeded twice background in the tester strains TA98, TA 100, TA102 examined at dose levels up to 5000 µg/plate in the absence of a metabolic activation source (S9) or in the presence of S9. Therefore, test substance was considered to be non-genotoxic (non-mutagenic) in Salmonella tester strains TA98, TA100, TA102, TA 1535 and TA 1537 under the conditions employed.

There was no evidence of an increase of revertant colonies after treatment with the test item.

Under the conditions of the study, the test substance was negative for mutagenic potential.