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Toxicological information

Genetic toxicity: in vitro

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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study report

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1987
Report Date:
1987

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
see Principles
Principles of method if other than guideline:
The strains subcultured from storage dishes have been cultivated at 37°C in a nutritive medium (oxoid No 2 CM67). They have been collected in the morning from the shelf (approx. 2 x 10 x e9 bacteria/ml) and used the same day after approx. 16 hours of culture. The minimum medium used to evi-dence the mutants His+ is the V.B. medium.
The spreadings have been made with an overlayer containing a concentra-tion of 0.6 .% agar oxoid, 0.5 % sodium chloride, 0.05 mM biotine and L- Histidine, for a total volume of 2.5 ml. The amount of histidine has been cal-culated so as to allow the bacteria to divide 2 or 3 times on the plates since some mutagene agents operate only on growing cells.
A 500 mg/kg dose of Araclar 1254 dissolved in maize oil has been injected to 5prague-Oawley rats, through the peritoneum. On the 5th day after injec-tion, the rats were slaughtered. Livers were taken and homogenized at 4 •C in 0,15 M potassium chloride, and the mix was centrifuged 20 minutes at 9000 g. After centrifugation, the supernatant which constitutes the 59 frac-tion was kept in liquid nitrogen in 2 ml fractions.
The product Lab 870 has been solubilized in water for injection (Aguettant, batch 7428) at the concentration of 20 mg/ml. Successive dilutions have been carried out in the same solvent at following concentrations 10 - 5 - 1 - 0.1 mg/ml. The solutions and the pure product have been kept at 5°C for the trial duration.
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
PRODUCT Lab 870 (Beta-Cyclodextrin)
LOT 392344

Method

Target gene:
not applicable
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Species / strain / cell type:
S. typhimurium TA 1538
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
Pretest: 0.01 - 0.05 - 0.1 - 0.5 - 1 - 2 -4 mg /plate
Test: 0.1 - 0.5 - 1 - 2 - 4 mg/plate.
Vehicle / solvent:
water for injection
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: amino-2-anthracene (2-anthramine)
Remarks:
all strains with S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
TA98, TA1538 without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
TA1537 without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
TA100 without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
ethylmethanesulphonate
Remarks:
TA1535 without S9
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation);

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3



DETERMINATION OF CYTOTOXICITY
- Method: mitotic index

Evaluation criteria:
None of the values obtained in presence of the product tested has been higher than, or equal to twice the value obtained in the presence of a soLvent with and without metabol ic activation on the 5 bacterial strains used.
Statistics:
not reported

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
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
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
not applicable
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: not reported
- Effects of osmolality: not reported
- Evaporation from medium: not reported
- Water solubility: not reported
- Precipitation: not reported
- Other confounding effects: not reported


RANGE-FINDING/SCREENING STUDIES: yes


COMPARISON WITH HISTORICAL CONTROL DATA: not reported


ADDITIONAL INFORMATION ON CYTOTOXICITY: not reported
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Substance is not mutagenic under the conditions used.
Executive summary:

The substance Beta-Cyclodextrin (LAB 870) has been tested on the 5 strains of Salmonella typhimurium (TA 98, TA 100, TA 1535, TA 1537, TA 1538) with and without metabolic activation (S9 -mix) according to the protocol of Ames and similar to OECD Guideline 471. A range of non-toxical concentrations had been determined in a preliminary experimentation on the strain TA 100 without metabolic activation. The 5 concentrations chosen (0.1, 0.5, 1, 2 and 4 mg/plate)have been tested in triplicate on the 5 strains quoted above, with and without metabolic activation. The results were confirmed in a second experiment separately from the first one.In each test a negative control (solvent) and a positive one (specific standard mutagene) had been included. The values obtained were consistent with the standards. Under these circumstances, the product did not show any mutagene power towards thestrains TA98, TA 100, TA 1535, TA 1537, TA .1538 with and without metabolic activation.