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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-report according to guideline.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Qualifier:
according to
Guideline:
EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
GLP compliance:
yes
Type of assay:
in vitro mammalian chromosome aberration test

Test material

Reference
Name:
Unnamed
Type:
Constituent

Method

Target gene:
not applicable
Species / strain
Species / strain / cell type:
lymphocytes: human, healthy donors without medication
Metabolic activation:
with and without
Metabolic activation system:
liver S9-mix of Aroclor 1254 induced rats
Test concentrations with justification for top dose:
0, 31.25, 62.5, 125, 250, 500 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The solvent was chosen according to its solubility properties and its non-toxicity for the cells.
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
DMSO 1% (v/v)
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
mitomycin C
Remarks:
without metabolic activation

Migrated to IUCLID6: 0.1 or 0.2 µg/mL
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
DMSO 1% (v/v)
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
with metabolic activation

Migrated to IUCLID6: 10 or 20 µg/ mL
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: 48 h
- Exposure duration: 4 h or 24 h without metabolic activation, 4 h with metabolic activation
- Fixation time (start of exposure up to fixation or harvest of cells): 22 h



SPINDLE INHIBITOR (cytogenetic assays): colcemid 0.9 µg/mL
STAIN (for cytogenetic assays): Giemsa stain


NUMBER OF REPLICATIONS: two cultures each two slide preparations


NUMBER OF CELLS EVALUATED: 100 metaphase per slide were evaluated


DETERMINATION OF CYTOTOXICITY
- Method: mitotic index


OTHER EXAMINATIONS:
- Determination of polyploidy: yes
Evaluation criteria:
Comparison of the number of chromosome aberrations of the samples with those of the solvent control. Total number of cells with aberrations exclusive of gap damage are analysed.
evaluation criteria:
The test is regarded to have a positive result, if the number of chromosomal aberrations is significantly increased (p<=0.05) compared with the solvent control, this increase is dose-dependent and both duplicate cultures lead to the same results. The increase should not occur in the severly cytotoxic range (mitotic index < 0.25).
Statistics:
Fisher-test (p<= 0.05)

Results and discussion

Test results
Species / strain:
lymphocytes: human, healthy donors without medication
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
125 µg/ mL 24 h exposure, 250 µg/ mL 4 h exposure
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Remarks on result:
other:
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Treatment

µg/mL

S9-mix

4 h exposure

24 h exposure

Mitotic index#

number of metaphases scored

% of cells with aberrations excluding gaps

Mitotic index#

number of metaphases scored

% of cells with aberrations excluding gaps

DMSO

-

-

1.00

200

1.5

1.00

200

2.0

test substance

31.25

-

-

-

-

0.86

200

2.5

62.5

-

0.92

200

1.5

0.55

200

3.5

125

-

1.27

200

1.5

0.47

164#

3.7

250

-

0.58

183#

4.5

0.28

35#

3.2

500

-

0.43

101#

5.2

-

-

-

Mitomycin C

0.2

-

0.62

200

12.0*

0.55

200

13.0*

the following concentrations were not evaluated:

31.25 µg/ mL (4 h exposure), 500 µg/ mL ( 24 h exposure) 0.1 µg mitomycin C (4 and 24 h exposure)

Treatment

µg/mL

S9-mix

4 h exposure

Mitotic index#

number of metaphases scored

% of cells with aberrations excluding gaps

DMSO

-

+

1.00

200

1.0

test substance

31.25

+

-

-

-

62.5

+

0.77

200

2.0

125

+

1.62

200

1.0

250

+

0.40

183#

4.5

500

+

0.13

11#

0.0

Cyclophosphamide

10

+

0.63

200

10.5*

the following concentrations were not evaluated:

31.25 µg/ mL, 20 µg/ mL cyclophosphamide

#          mitotic index: number of metaphases/ 1000 cells: negative control = 1.00
##        no more metaphase of sufficient quality for evaluation due to cytotoxicity of Triallylcyanurate
*          significantly different from negative control (p=0.05)

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative