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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

No indication of mutagenicity in Ames test.

Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
July - September 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
21 July 1997
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Details on mammalian cell type (if applicable):
CELLS USED
- Source of cells: Molecular Toxicology Inc., 157 Industrial Park Dr. Boone, NC 28607, U.S.A.
- Suitability of cells: standard cells, genotype confirmed by the laboratory

Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254 induced rat liver S9 fraction
Test concentrations with justification for top dose:
Based on initial experiment: 156.25, 312.5, 625, 1250 , 2500, and 5000 µg/plate both in the absence and presence of metabolic activation system (10% v/v S9 mix) for all the tester strains.
In the initial experiment, normal bacterial lawn was observed up to the tested concentration of 5000 µg/plate both in the absence and presence of the metabolic activation system (5% v/v S9 mix) in the tester strains of TA1537, TA1535, TA98, TA100 and TA102.
Vehicle / solvent:
dimethyl sulfoxide (DMSO)
Untreated negative controls:
yes
Remarks:
DMSO
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
mitomycin C
other: 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Incubation period: 48 hrs
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):

SELECTION AGENT (mutation assays): crystal violet for rfa wall mutation, biotin for growth requirement (except TA 102), UV exposure sensitivity (except for TA102), histidine for growth requirement, resistance to ampicillin (TA98, 100, 102) for pKM101 plasmid, resistance to tetracycline (TA102) for pAQ1 plasmid

NUMBER OF REPLICATIONS: 3 per each concentration

NUMBER OF CELLS EVALUATED: plate density 1 - 2 x 10^9 bacteria/mL


DETERMINATION OF CYTOTOXICITY : decrease in the number of revertant colonies per plate and/or by a thinning of the bacterial background lawn
Rationale for test conditions:
Guideline requirements
Evaluation criteria:
A result is considered positive if a concentration-related increase over the range tested and/or a reproducible increase at one or more concentrations in the number of revertant colonies per plate in at least one strain with or without metabolic activation system.

Strains TA1535, TA1537
Data sets were judged positive, if the increase in mean revertants at the peak of the dose response was equal to or greater than 3.0 times the mean negative control value.

Strain TA98, TA100 and TA102
Data sets were judged positive, if the increase in mean revertants at the peak of the dose response was equal to or greater than 2.0 times the mean negative control value.

Statistical analysis was used as an aid in the evaluation of the dose response.
A response that did not meet all three of the above criteria (magnitude, concentration-responsiveness, reproducibility) was not judged as positive.
Negative results obtained in the initial toxicity-mutation test were confirmed by a second trial, using the same method as specified above, with an alteration in concentration spacing
Statistics:
Simple linear regression analysis was performed for TA1537, TA1535, TA98, TA100 and TA102, separately, to assess the dose dependent nature of any increase in revertant colonies.
Key result
Species / strain:
S. typhimurium TA 1535
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
Key result
Species / strain:
S. typhimurium TA 1537
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
Key result
Species / strain:
S. typhimurium TA 98
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
Key result
Species / strain:
S. typhimurium 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
Key result
Species / strain:
S. typhimurium TA 102
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:
Normal bacterial background lawn was observed up to the tested concentration of 5000 µg/plate both in the absence and presence of the metabolic activation system (10% v/v S9 mix) in tester strains. Results revealed that there was no positive mutagenic effect in tester strains TA1537, TA1535, TA98, TA100, and TA102, at the tested concentration levels 156.25, 312.5, 625, 1250, 2500, and 5000 µg/plate of ethyl vanillin glucoside, in the absence and presence of the metabolic activation system (10% v/v S9 mix), when compared with the concurrent negative control. Statistical analysis did not reveal any significant effect.
Conclusions:
From results of this study, it is concluded that ethyl vanillin glucoside is non-mutagenic to any strains of Salmonella typhimurium , viz., TA1537, TA1535, TA98, TA100, and TA102 when tested under the specified experimental conditions.
Executive summary:

From results of this study, it is concluded that ethyl vanillin glucoside is non-mutagenic to any strains of Salmonella typhimurium , viz., TA1537, TA1535, TA98, TA100, and TA102 when tested under the specified experimental conditions.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification