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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
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report Date:
1993

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
other: In Vitro Mammalian Chromosome Aberration Test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
- Lot/batch No.: 12-208-222

Method

Species / strain
Species / strain / cell type:
lymphocytes: human
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
Experiment I:
without S9 mix (exposure period 24 h and 48 h)
24h: 10, 30, 100 µg/mL
48h: 100 µg/mL
with S9 mix (exposure period 4 h)
24h: 300, 600, 1680 µg/mL
48h: 1680 µg/mL

Experiment II
without S9 mix (exposure period 24 h and 48 h)
24h: 30, 100, 150 µg/mL
48h: 200 µg/mL
with S9 mix (exposure period 4 h)
24h: 500, 1000, 1680 µg/mL
48h: 1680 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
ethylmethanesulphonate
Remarks:
without S9 mix (0.30 mg/mL)
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
with S9 mix (30 µg/mL)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 4, 24, 48 hours
- Fixation time (start of exposure up to fixation or harvest of cells): 3 hours before harvesting

SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): Giemsa

NUMBER OF REPLICATIONS: 2

NUMBER OF CELLS EVALUATED: 200 per dose

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index
Statistics:
Statistical significance at the five per cent level (p < 0.05) was evaluated by means of the chi-square test. Evaluation was performed only for cells carrying aberrations exclusive gaps.

Results and discussion

Test results
Key result
Species / strain:
lymphocytes: human
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid

Any other information on results incl. tables

Experiment I (exposure period 24 h)

fixation interval 24 h after start of the treatment

Test groups

S9 mix

Analyses of chromosomes

Incl. Gaps %

Excl. gaps %

Negative control

-

2.00

1.00

1% Solvent control DMSO

-

1.50

0.50

0.33 mg/ml Positive contol

-

16.00

13.0

10 µg/ml Test article

-

1.00

1.0

30 µg/ml Test article

-

0.00

0.00

100 µg/ml Test article

-

1.00

0.50

 

Experiment I (exposure period 48 h)

fixation interval 48 h after start of the treatment

Test groups

S9 mix

Analyses of chromosomes

Incl. Gaps %

Excl. gaps %

1% Solvent control DMSO

-

1.50

1.00

100 µg/ml Test article

-

1.50

0.50

 

Experiment II (exposure period 24 h)

fixation interval 24 h after start of the treatment

Test groups

S9 mix

Analyses of chromosomes

Incl. Gaps %

Excl. gaps %

Negative control

-

3.50

1.50

1.0% Solvent control DMSO

-

2.00

1.50

0.33 mg/ml Positive contol

-

14.50

10.00

30 µg/ml Test article

-

2.00

1.00

100 µg/ml Test article

-

1.50

0.50

150 µg/ml Test article

-

1.00

0.50

 

Experiment II (exposure period 48 h)

fixation interval 48 h after start of the treatment

Test groups

S9 mix

Analyses of chromosomes

Incl. Gaps %

Excl. gaps %

1.0% Solvent control DMSO

-

0.00

0.00

150 µg/ml Test article

-

3.00

2.00

 

 

 

 

 

 

Experiment I (exposure period 4 h)

fixation interval 24 h after start of the treatment

Test groups

S9 mix

Analyses of chromosomes

Incl. Gaps %

Excl. gaps %

Negative control

+

0.50

0.00

1.0% Solvent control DMSO

+

0.50

0.50

30.0 µg/ml Positive contol CPA

+

17.00

14.00

300 µg/ml Test article

+

0.50

0.00

600 µg/ml Test article

+

2.00

1.00

1680 µg/ml Test article

+

1.00

0.50

 

Experiment I (exposure period 4 h)

fixation interval 48 h after start of the treatment

Test groups

S9 mix

Analyses of chromosomes

Incl. Gaps %

Excl. gaps %

1.0% Solvent control DMSO

+

1.00

0.50

1680 µg/ml Test article

+

0.00

0.00

 

Experiment II (exposure period 4 h)

fixation interval 24 h after start of the treatment

Test groups

S9 mix

Analyses of chromosomes

Incl. Gaps %

Excl. gaps %

Negative control

+

2.00

1.00

1.0% Solvent control DMSO

+

1.50

0.00

30.0 µg/ml Positive contol CPA

+

10.50

10.00

500 µg/ml Test article

+

1.00

0.00

1000 µg/ml Test article

+

1.00

0.50

1680 µg/ml Test article

+

0.00

0.00

 

Experiment II (exposure period 4 h)

fixation interval 48 h after start of the treatment

Test groups

S9 mix

Analyses of chromosomes

Incl. Gaps %

Excl. gaps %

1.0% Solvent control DMSO

+

0.50

0.50

1680 µg/ml Test article

+

2.50

1.50

 

Applicant's summary and conclusion