Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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
Study period:
07 May 2003 - 10 Sep 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to the appropriate OECD test guideline and in compliance with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003
Report date:
2003

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: requirements of UK Department of Health Committee on Mutagenicity Guidelines for the Mutagenicity Testing of Chemicals
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
The Department of Health of the Government of the United Kingdom
Type of assay:
in vitro mammalian chromosome aberration test

Test material

Constituent 1
Reference substance name:
Amides, C18, branched and linear
Molecular formula:
Not applicable as UVCB
IUPAC Name:
Amides, C18, branched and linear
Test material form:
semi-solid (amorphous): gel
Remarks:
migrated information: paste
Details on test material:
- Name of test material (as cited in study report): PERFAD FM 3337
- Physical state: cream coloured paste
- Analytical purity: 95%
- Lot/batch No.: 607459
- Storage condition of test material: at room temperature protected from light

Method

Target gene:
Not applicable.
Species / strain
Species / strain / cell type:
lymphocytes: of fresh heparinised whole blood from peripheral circulation of a volunteer
Details on mammalian cell type (if applicable):
- Type and identity of media: Eagle's minimal essential medium with HEPES buffer (MEM), supplemented with L-glutamine, penicillin/streptomycin, amphotericin B, and 15% foetal calf serum (FCS).
- Properly maintained: yes
- The lymphocytes were stimulated to divide by the addition of phytohaemagglutinin (PHA) at 90 µg/mL final concentration.
Additional strain / cell type characteristics:
other: The volunteer donor had not been exposed to high levels of radiation or hazardous chemicals and had not knowingly recently suffered from a viral infection.
Metabolic activation:
with and without
Metabolic activation system:
co-factor supplemented S9-Mix from male Sprague-Dawley rats induced with phenobarbitone (80 mg/kg bw) and ß-naphtoflavone (100 mg/kg bw); Experiment 1: 2% final concentration and Experiment 2: 1 % final concentration
Test concentrations with justification for top dose:
- Preliminary Toxicity Test: 19.53, 39.06, 78.13, 156.25, 312.5, 625, 1250, 2500, and 5000 µg/mL
- Experiment 1: 9.77, 19.53, 39.06, 78.13, 156.25, and 312.5 µg/mL (-/+ S9)
- Experiment 2: 9.77, 19.53, 39.06, 78.13, 156.25, and 312.5 µg/mL (-S9); 4.88, 9.77, 19.53, 39.06, 58.60, and 78.13 µg/mL (+S9)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
cyclophosphamide
mitomycin C
Remarks:
-S9: Mitomycin C (MMC) 0.4 and 0.2 µg/mL in MEM; +S9: cyclophosphamide (CP) 10 µg/mL in DMSO
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: none
- Exposure duration: 4 h (Experiment 1 and 2, +S9 and Experiment 2 +S9); 24 h ( Experiment 2 -S9)
- Expression time (cells in growth medium): 20 h (only Experiment 1 and 2, +S9 and Experiment 2 +S9)
- Fixation time (start of exposure up to fixation or harvest of cells): 24 h

SPINDLE INHIBITOR (cytogenetic assays): colcemid 0.1 µg/mL
STAIN (for cytogenetic assays): 5% Gurrs Giemsa

NUMBER OF REPLICATIONS: 2 independent experiments

NUMBER OF CELLS EVALUATED: Mitotic Index: 2000 lymphocyte cell nuclei; aberrations: 100 consecutive well-spread metaphases from each culture were counted (termination at 50 metaphases in case there were approximately 50% cells with aberrations)

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index

OTHER EXAMINATIONS:
- Determination of polyploidy: yes
Evaluation criteria:
A positive response was recorded for a particular treatment if the % cells with aberrations, excluding gaps, markedly exceed that seen in the concurrent control, either with or without a clear dose response relationship. For modest increase in aberration frequency a dose response relationship is generally required and appropriate statistical tests may be applied in order to record a positive response.
Statistics:
The frequency of cells with aberrations excluding gaps and the frequency of polyploid cells was compared, where necesary, with the concurrent vehicle control value using Fisher's Exact Test.

Results and discussion

Test results
Species / strain:
lymphocytes:
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Experiment 1: +S9 at 156.25 µg/mL, -S9 at 78.13 µg/mL; Experiment 2: +S9 at 39.06 µg/mL, - S9 at 19.53 µg/mL
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no
- Effects of osmolality: no
- Precipitation: yes (at 156.25 µg/mL and above)

COMPARISON WITH HISTORICAL CONTROL DATA: The positive and vehicle control data are within the range of the laboratory's historical control data.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Tab. 1: Experiment 1: 4 h treatment, 24 h fixation - Without Metabolic Activation

Concentration

Mitotic index [%]

Polyploid cells

Aberrant cells

incl. gaps

excl. gaps

Solvent control

100

0.0

0.5

0.5

9.77 µg/mL

96

0.0

2.5

2.0

19.53 µg/mL

126

0.0

2.0

0.5

39.06 µg/mL

66

2.0

3.0

1.5

MMC 0.4 µg/mL

34

0.0

44.7

38.0

MMC: mitomycin C; solvent control: DMSO

 

Tab. 2: Experiment 1: 4 h treatment, 24 h fixation - With Metabolic Activation (2% S9-Mix)

Concentration

Mitotic index [%]

Polyploid cells

Aberrant cells

incl. gaps

excl. gaps

Solvent control

100

0.5

1.0

0.5

19.53 µg/mL

110

1.0

1.5

0.5

39.06 µg/mL

85

0.0

0.5

0.5

78.13 µg/mL

72

0.0

0.5

0.5

CP 10 µg/mL

15

0.5

29.5

22.0

CP: cyclophosphamide; solvent control: DMSO

 

Tab. 3: Experiment 2: 24 h treatment, 24 h fixation - Without Metabolic Activation

Concentration

Mitotic index [%]

Polyploid cells

Aberrant cells

incl. gaps

excl. gaps

Solvent control

100

0.0

4.0

2.0

4.88 µg/mL

100

0.0

0.0

0.0

9.77 µg/mL

73

0.0

2.0

0.5

19.53 µg/mL

48

0.0

1.1

0.0

MMC 0.2 µg/mL

26

0.0

55.0

45.0

MMC: mitomycin C; solvent control: DMSO

 

Tab. 4: Experiment 2: 4 h treatment, 24 h fixation - With Metabolic Activation (1% S9-Mix)

Concentration

Mitotic index [%]

Polyploid cells

Aberrant cells

incl. gaps

excl. gaps

Solvent control

100

0.0

1.5

1.0

9.77 µg/mL

138

1.0

4.0

2.0

19.53 µg/mL

111

0.0

1.0

1.0

39.06 µg/mL

134

0.5

3.5

1.0

CP 10 µg/mL

47

0.0

48.0

36.0

CP: cyclophosphamide; solvent control: DMSO

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative