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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP compliant OECD Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
3013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Version / remarks:
1997
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:
EPA OPPTS 870.5375 - In vitro Mammalian Chromosome Aberration Test
Deviations:
no
GLP compliance:
yes
Remarks:
Hessischens Ministerium für Umwelt, Energie, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany
Type of assay:
in vitro mammalian chromosome aberration test

Test material

Constituent 1
Reference substance name:
70955-07-6
EC Number:
615-218-7
Cas Number:
70955-07-6
IUPAC Name:
70955-07-6
Details on test material:
- Name of test material (as cited in study report): Alcohols, tallow, propoxylated (>1<2.5 mol PO)
- Physical state: lowly viscous fluid, yellowish, clear
- Analytical purity: 96.6%
- Lot/batch No.: CP11090009
- Expiration date of the lot/batch: April 18, 2013
- Storage condition of test material: Room temperature, avoid temperatures > 40°C

Method

Target gene:
not applicable
Species / strain
Species / strain / cell type:
Chinese hamster lung fibroblasts (V79)
Details on mammalian cell type (if applicable):
Species/strain: lymphocytes: cultured peripheral human lymphocytes
Details on mammalian cell lines:
- Type and identity of media:
MEM medium (minimal essential medium) containing Hank’s salts,
glutamine and Hepes (25 mM) with glutamine supplemented with
- 10%(v/v) fetal bovine serum (FBS)
- 1% (v/v) penicillin/streptomycin (100 U/mL/100 Q g/mL)
During exposure to the test substance (4-hour treatment), MEM medium was used
without FBS supplementation.
- Properly maintained: yes/no
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: no
Metabolic activation:
with and without
Metabolic activation system:
cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with phenobarbital/beta-naphtoflavone
Test concentrations with justification for top dose:
4 h treatment (IA), without S9 mix: 20.3, 40.6, 81.3, 162.5, 325.0 (Phase separation = PS), 650.0 PS, 1300.0 PS, 2600.0 PS, 5200 PS
18 h treatment (IIA) without S9 mix: 0.8, 1.6, 3.1, 6.3, 12.5, 25.0, 50.0, 100.0 PS, 200.0 PS
28 h treatment (IIA) without S9 mix: 0.8, 1.6, 3.1, 6.3, 12.5, 25.0, 50.0, 100.0 PS, 200.0 PS
4 h treatment (IA), with S9 mix: 20.3, 40.6, 81.3, 162.5, 325.0, 650.0, 1300.0 PS, 2600.0 PS, 5.2 PS
4 h treatment (IB) with S9 mix: 75.0, 150.0, 300.0 PS, 600.0 PS, 1000.0 PS, 1400.0 PS, 1800.0, 2000.0 PS, 2600.0 PS
4 h treatment (IC) with S9 mix75.0, 150.0, 300.0 PS, 600.0 PS, 1000.0 PS, 1400.0 PS, 1800.0, 2000.0 PS, 2600.0 PS
4 h treatment (IIB) with S9 mix: 30.0, 60.0, 80.0, 100.0, 120.0, 140.0, 160.0, 180.0, 200.0



Vehicle / solvent:
- Vehicle used: ethanol
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
Ethanol
True negative controls:
no
Positive controls:
yes
Positive control substance:
ethylmethanesulphonate
Remarks:
1000 µg/mL (Exp IA) and 600 µg/mL (Exp. IIA), + S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
2.0 µg/mL (Exp. IB), 1.4 µg/mL (Exp. IC), and 2.4 µg/mL (Exp. IIA and IIB), -S9
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4, 18 and 28 h
- Fixation time (start of exposure up to fixation or harvest of cells): 4h treatment: 18 and 28 h. 18 h treatment. 28 h treatment: 28 h
SPINDLE INHIBITOR (cytogenetic assays): Colcemid 0.2 µg/mL in medium
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: two replications each in 5 independent experiments
NUMBER OF CELLS EVALUATED: 100 per culture, except for the positive controls in Experiment IIA and IIB after 28 h preparation interval with metabolic activation, where only 50 metaphases were evaluated
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index
OTHER EXAMINATIONS:
- Determination of polyploidy: yes
Evaluation criteria:
A test substance was considered positive (clastogenic) in the chromosome aberration test if:
a) It induced a dose-related or statistically significant (Fisher´s exact test, p < 0.05) increase in the number of cells with chromosome aberrations.
b) The number of induced structural chromosome aberrations is not in the range of laboratory control historical data. However, both biological and statistical significance should be considered together.
A test substance was considered negative (not clastogenic) in the chromosome aberration test if no significant increase in the number of cells with chromosome is observed and if the number of induced structural chromosome aberration in all evaluated dose groups is in the range of the laboratory historical control data.
Statistics:
Fisher´s exact test, p < 0.05

Results and discussion

Test results
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
At the highest evaluated concentration in experiment IA (-S9 mix) and in experiment IC (+S9 mix) the mitotic index was 49.7, 45.3 % of control, respectively
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1 Summary of the chromosome aberration study without metabolic activation

Exp

Preparation interval

Test item concentration (µg/mL)

Polyploid cells in %

Endomitotic

cells in %

Cell numbers in % of control

Mitotic indices in % of control

Aberrant cells in %

Incl. gaps*

excl. gaps*

With exchanges

Exposure period 4h without S9 mix

IA

 

 

 

 

18 h

 

 

 

 

Solvent control 1

2.9

0.0

100.0

100.0

3.0

2.5

0.0

Positive control 2

n.d.

n.d.

n.d.

80.0

17.0

17.0s

11.0

20.3

3.8

0.0

100.2

96.4

1.5

0.5

0.5

40.6

2.5

0.0

79.6

95.2

1.0

1.0

0.0

81.3

2.2

0.0

73.4

49.7

1.5

1.5

0.0

Exposure period 18h without S9 mix

II A

 

 

 

 

18 h

 

 

 

 

Solvent control 1

2.1

0.0

100.0

100.0

4.0

3.5

0.5

Positive control 3

n.d.

n.d.

n.d.

66.1

29.0

28.5s

14.0

1.6

2.2

0.0

87.4

86.0

0.5

0.5

0.0

3.1

2.3

0.0

90.6

88.3

2.0

1.0

0.0

6.3

2.4

0.0

103.0

95.1

1.0

0.5

0.0

Exposure period 28h without S9 mix

II A

 

 

 

 

28 h

 

 

 

 

Solvent control 1

3.0

0.0

100.0

100.0

0.0

0.0

0.0

Positive control 3

n.d.

n.d.

n.d.

66.6

34.5

34.5s

25.5

1.6

2.6

0.0

110.1

86.5

3.5

2.0

0.0

3.1

2.5

0.0

91.6

82.6

1.5

0.5

0.0

6.3

1.3

0.0

91.8

68.0

0.0

0.0

0.0

* Including cells carrying exchanges

n.d. Not determined

S Aberration frequency statistically higher than corresponding control values

1 Ethanol 0.5% (v/v)

2 EMS 1000.0 µg/mL

3 EMS 600.0 µg/mL

Table 2 Summary of the chromosome aberration study with metabolic activation

Exp

Preparation interval

Test item concentration (µg/mL)

Polyploid

cells in %

Endomitotic

cells in %

Cell numbers in % of control

Mitotic indices in % of control

Aberrant cells in %

Incl. gaps*

excl. gaps*

With exchanges

Exposure period 4 hours with S9 mix

IB

 

 

 

 

18 h

 

 

 

 

Solvent control 1

1.9

0.0

100.0

100.0

3.5

2.0

0.0

Positive control 2

n.d.

n.d.

n.d

60.7

26.0

26.0s

10.5

75.0

1.1

0.0

116.5

106.5

3.0

3.0

1.5

150.0

0.9

0.0

87.0

82.1

2.0

2.0

0.5

300.0

1.2

0.0

96.9

66.3

3.0

3.0

1.0

IC

 

 

 

 

18 h

 

 

 

 

Solvent control 1

2.3

0.0

100.0

100.0

2.0

2.0

1.5

Positive control 3

n.d.

n.d.

n.d.

59.6

23.5

23.5s

14.5

150.0

2.9

0.0

85.8

109.1

2.0

1.5

0.0

300.0

2.9

0.0

77.4

80.5

1.5

1.5

1.0

350.0

1.4

0.0

77.0

45.3

2.0

2.0

1.0

IIA

 

 

 

 

28 h

 

 

 

 

Solvent control 1

1.3

0.0

100.0

100.0

2.5

2.0

0.5

Positive control 4#

n.d.

n.d.

n.d.

109.1

54.0

54.0S

24.0

25.0

2.5

0.0

76.1

99.7

1.5

1.5

0.5

50.0

1.9

0.0

106.2

106.1

3.5

3.0

0.0

100.0

1.7

0.0

90.6

68.5

2.5

1.5

0.0

IIB

 

 

 

 

28 h

 

 

 

 

Solvent control 1

2.3

0.0

100.0

100.0

2.5

2.5

0.0

Positive control 4#

n.d

n.d.

n.d.

107.7

54.0

53.0s

22.0

120.0

1.5

0.0

106.1

97.8

3.0

2.5

1.0

160.0

2.9

0.0

75.9

101.9

2.0

1.5

0.5

200.0

3.0

0.0

74.0

56.9

0.5

0.5

0.0

* Including cells carrying exchanges

n.d. Not determined

S Aberration frequency statistically higher than corresponding control values

PS Phase separation occurred at the end of treatment

# 50 metaphases per culture were evaluated

1 Ethanol 0.5% (v/v)

2 CPA 2.0 µg/mL

3 CPA 1.4 µg/mL

4 CPA 2.4 µg/mL

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative