L-gluco-Octitol, 1,5-anhydro-6,8-dideoxy-, (7.xi.)-

Genetic toxicity: in vitro

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:

23 November - 16 December 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP study conducted according to OECD Guideline 471 with minor deviations: no data on number of cells per culture

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

no data on number of cells per culture

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material: Mexoryl SBF
- Physical state: Thick brownish paste (very sticky)
- Analytical purity: 92 %
- Lot/batch No.: 003D001
- Date of receipt: 18 November 2005
- Expiration date of the lot/batch: October 2006
- Storage condition of test material: Stored at +4 °C and protected from light

Method

Target gene:

His+ for S. typhimurium

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

other: see table 7.6.1/1

Metabolic activation:

with and without

Metabolic activation system:

10 % (v/v) S9 mix; S9 fraction prepared from liver homogenates of rats induced with Aroclor 1254 (500 mg/kg bw) by intraperitoneal route

Test concentrations with justification for top dose:

Preliminary toxicity test: 10, 100, 500, 1000, 2500 and 5000 µg/plate, with and without S9 mix in TA 98, TA 100 and TA 102 strains (direct plate incorporation method)
Mutagenicity tests:
- Experiment 1: 312.5, 625, 1250, 2500 and 5000 μg/plate, with and without S9 mix in all 5 strains (direct plate incorporation method)
- Experiment 2: 312.5, 625, 1250, 2500 and 5000 μg/plate, with S9 mix (preincubation method) and without S9 mix (direct plate incorporation method) in all 5 strains

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Water for injections
- Batch No.: GVD21A
- Source: Fresenius-Kabi, Sèvres, France
Formulation procedure:
- Prior to use, the test item was maintained at room temperature for 2-3 h, before manipulation.
- Test item was dissolved in the vehicle at a concentration of 50 mg/mL for the preliminary toxicity test and both mutagenicity experiments. The preparations were made immediately before use.

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water for injections

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: sodium azide (1 µg/plate in TA 100 and TA 1535); 9-aminoacridine (50 µg/plate in TA 1537); 2-nitrofluorene (0.5 µg/plate in TA 98); mitomycin C (0.5 µg/plate in TA 102)

Remarks:

without metabolic activation

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water for injections

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Anthramine (2 µg/plate in TA 100, TA 1535, TA 1537 and TA 98); 2-Anthramine (10 µg/plate in TA 102)

Remarks:

with metabolic activation

Details on test system and experimental conditions:

SOURCE OF TEST SYSTEM: All five strains of Salmonella typhimurium were supplied by B.N. Ames' Laboratory (University of California, Berkeley, USA).

METHOD OF APPLICATION: In agar (direct plate incorporation and preincubation method)

DURATION
- Preincubation period: 60 minutes at 37 °C
- Incubation period: 48-72 h at 37 °C for both direct plate incorporation and preincubation methods

NUMBER OF REPLICATIONS:
- Preliminary experiment: 1 plate/dose
- Mutagenicity experiments: 3 plates/dose

DETERMINATION OF CYTOTOXICITY
- Method: Evaluation of the toxicity was performed on the basis of the observation of the decrease in the number of revertant colonies and/or a thinning of the bacterial lawn.

OTHER: After 48 to 72 h of incubation at 37 °C, revertants were scored with an automatic counter (Cardinal counter, Perceptive Instruments, Suffolk, UK).

Evaluation criteria:

- A reproducible 2-fold increase (for the TA 98, TA 100 and TA 102 strains) or 3-fold increase (for the TA 1535 and TA 1537 strains) in the number of revertants compared with the vehicle controls, in any strain at any dose-level and/or evidence of a dose-relationship was considered as a positive result.
- Reference to historical data along with biological relevance was considered during the evaluation.

Statistics:

None

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: Test substance was freely soluble in the vehicle at 50 mg/L and non toxic at the doses used (10, 100, 500, 1000, 2500 and 5000 µg/plate).

MUTAGENICITY TESTS:
- No precipitate was observed in the petri plates when scoring the revertants at any dose-level.
- No toxicity was noted towards all the strains used, both with and without S9 mix.
- Test item did not induce any noteworthy increase in the number of revertants, both with and without S9 mix, in any of the five strains.

OTHERS:
- Sterility of the S9 mix was checked before the beginning and at the end of each experiment and was found to be satisfactory.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

See file attached

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

Under the test conditions, Mexoryl SBF is not considered as mutagenic in S. typhimurium strains (TA 1535, TA 1537, TA 98, TA 100 and TA 102).

Executive summary:

In a reverse gene mutation assay in bacteria, performed according to the OECD Guideline 471 and in compliance with GLP, strains of Salmonella typhimurium (TA 1535, TA 1537, TA 98, TA 100 and TA 102) were exposed to Mexoryl SBF at the following concentrations:

Preliminary toxicity test: 10, 100, 500, 1000, 2500 and 5000 µg/plate, with and without S9 mix in TA 98, TA 100 and TA 102 strains (direct plate incorporation method).

Mutagenicity tests:

- Experiment 1: 312.5, 625, 1250, 2500 and 5000 μg/plate, with and without S9 mix in all 5 strains (direct plate incorporation method).

- Experiment 2: 312.5, 625, 1250, 2500 and 5000 μg/plate, with S9 mix (preincubation method) and without S9 mix (direct plate incorporation method) in all 5 strains.

Metabolic activation system used in this test was10 % (v/v) S9 mix. S9 fraction was prepared from liver homogenates of rats induced with Aroclor 1254 (500 mg/kg bw) by intraperitoneal route. Vehicle control and positive control groups were also included in mutagenicity tests.

The test substance did not induce precipitation or toxicity at any of the doses used either in the preliminary toxicity test or mutagenicity tests. The positive and vehicle controls induced the appropriate responses in the corresponding strains. Mexoryl SBF showed no substantial increases in revertant colony numbers over control count obtained with any of the tested strains at any concentrations in either presence or absence of S9 mix

Under the test conditions, Mexoryl SBF is not considered as mutagenic in this bacterial system.