Fatty acids C16-18 (even numbered), mono, di and tri esters with sucrose

Genetic toxicity: in vitro

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

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

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 results

Test results 1

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

Test results 2

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

Test results 3

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

Test results 4

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

Test results 5

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.