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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
October 20-23, 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature, protected from light.
- Stability under test conditions: Expected to be stable for duration of testing.
- Solubility and stability of the test substance in the solvent/vehicle: Precipitation was noted at the 1580 and 5000 µg/plate concentrations.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test substance was ground, then made into solutions for the appropriate concentrations.
- Final preparation of a solid: Solutions were ground, vortexed, and sonicated in a water bath prior to use.

FORM AS APPLIED IN THE TEST (if different from that of starting material): Solution

Method

Target gene:
his, rfa, uvrA/B, R-factor, trp
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
chemically-induced rat liver S9
Test concentrations with justification for top dose:
1.58, 5.0, 15.8, 50, 158, 500, 1580, 5000 µg/plate
The top dose was the OECD standard limit dose.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
methylmethanesulfonate
other: ICR 191 Acridine 10 µg/mL, Daunomycin 60 µg/mL, 2-Aminoanthracene 100 µg/mL
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)
- Cell density at seeding (if applicable): 1 x 10^9 bacteria/mL

DURATION
- Preincubation period: 30 minutes (confirmatory test)
- Exposure duration: 65 hrs
- Expression time (cells in growth medium): 65 hrs

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth
Rationale for test conditions:
OECD guideline
Evaluation criteria:
Validity: Vehicle controls should appear normal with mean revertant colony counts should be close to historical controls or published values. Positive controls should produce a substantial increase in revertant colonies.
Toxicity: Partial or complete absence of background lawn of non-revertant bacteria, or a substantial dose-related reduction in revertant colony counts.
Mutagenicity: Mutation factor 2 or greater for Salmonella strains TA98, TA100, and E. coli strain WP2 uvrA; mutation factor 3 or greater for Salmonella strains TA1535 and 1537. These increases must be dose-related and/or reproducible.
Statistics:
Means and standard deviations.

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating 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
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, but tested up to precipitating 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:
cytotoxicity
Remarks:
500 ug/plate and above
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation was seen in the some tests at the 1580 and 5000 µg/plate dose levels.
- Other confounding effects: Contamination was also observed in some tests at concentrations of 158 µg/plate and above.



Applicant's summary and conclusion

Conclusions:
The test substance in not mutagenic to bacteria.
Executive summary:

The test substance Sucrose Palmitate Stearate MDT Grade was tested for mutagenicity in the Ames test according to OECD Guideline 437. The strains Salmonella TA98, TA100, TA1535, and TA1537, in addition to E. coli strain WP2 uvrA were tested with and without metabolic activation to doses of 1.58, 5, 15.8, 50, 158, 500, 1580, and 5000 µg/plate. Negative controls and positive controls were also tested. The results showed the test substance is not mutagenic to bacteria.