Fatty acids, tall-oil, iron salts

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 June 2001 - 26 September 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Justification for read-across, see attached file.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

GLP compliance:

yes (incl. QA statement)

Type of assay:

other: Testing and evaluation of clastogenic potential - chromosomal aberrations assays.

Test material

Test material information

Constituent 1

Reference substance name:

Tall oil

EC Number:

232-304-6

EC Name:

Tall oil

Cas Number:

8002-26-4

Molecular formula:

Not applicable - UVCB substance

Test material form:

liquid: viscous

Method

Species / strain

Species / strain / cell type:

Chinese hamster Ovary (CHO)

Details on mammalian cell type (if applicable):

The cell line used was Chinese hamster ovary (CHO) cells, obtained from the University of Leiden in 1987.

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced S9 enzymes (supernatant post-mitochondrial fraction obtained after centrifugation at 9000 G) were prepared from the livers of adult, male Fischer rats, as described by Ames et al (1975).

Test concentrations with justification for top dose:

For the first experiment, 9 dose levels, covering a wide concentration range, were tested. The highest dose was 5000 µg.ml-1, the maximum allowable concentration and subsequent dose levels were halving dilutions.

The presence of S9 mix had to be repeated due to toxicity and the following concentrations were tested:

in Test 1: 10, 20, 40, 60 and 80 µg.ml-1

In Test 2, the dose levels selected were:
Presence of S9 mix: 5, 10, 20, 30, 40, 50 and 60 µg.ml-1
Absence of S9 mix: 50, 55, 60, 62.5, 65, 67.5 and 70 µg.ml-1

Vehicle / solvent:

Dimethylsulphoxide

Controls

Untreated negative controls:

other: Historically negative control data

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

cyclophosphamide

methylmethanesulfonate

Results and discussion

Test results

Species / strain:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Genotoxicity:

other: weakly clastogenic

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

other: Historical negative control

Positive controls validity:

valid

Additional information on results:

Tall Oil, CAS No. 8002-26-4, was toxic to Chinese hamster ovary cells in vitro in both the presence and absence of S9 mix. It was tested to the maximum permitted concentration of 5000 µg.ml-1 in the absence of S9 mix and up to 80 µg.ml-1 in the presence of S9 mix (Test 1). Toxicity was noted at 60 and 80 µg.ml-1 in the presence of S9 mix and at 156-5000 µg.ml-1 in the absence of S9 mix. In Test 2, Tall Oil, CAS No. 8002-26-4, was tested up to 60 µg.ml-1 in the presence of S9 mix and up to 70 µg.ml-1 in the absence of S9 mix. Toxicity was noted in cultures treated with 30-60 µg.ml-1 (presence of S9 mix) and in cultures treated with 70 µg.ml-1 (absence of S9 mix).
It was concluded that Tall Oil, CAS No. 8002-26-4, was weakly clastogenic when tested with Chinese hamster ovary cell in vitro in the presence of S9 mix. The response was observed only at a concentration level deemed overtly toxic to the cells.

Applicant's summary and conclusion

Conclusions:

It was concluded that Tall Oil, CAS No. 8002-26-4, was weakly clastogenic when tested with Chinese hamster ovary cell in vitro in the presence of S9 mix.  The response was observed only at a concentration level deemed overtly toxic to the cells.

Executive summary:

Aim:

Tall Oil, CAS No. 8002-26-4, was submitted for testing and evaluation of clastogenic potential.  

Method:

Chromosomal aberrations assays were performed with duplicate, Chinese hamster ovary (CHO) cell cultures.  This study was conducted incorporating two independent tests.  Dimethylsulphoxide was the vehicle and cyclophosphamide and methyl methanesulphonate were the positive controls used in both tests. Both tests were conducted in the presence and absence of a postmitochondrial supernatant fraction obtained from the livers of adult, male rats treated with Aroclor 1254 (S9) and a NADPH-generating system.

Cultures, established approximately 20 hours before testing, were treated for 6 h in the presence or 6 and 22 hours in the absence of S9 mix.  Cultures were harvested at 24 h (Test 1 and 2) or 48 h (Test 2) post treatment.

Results/Conclusion

Tall Oil, CAS No. 8002-26-4, was toxic to Chinese hamster ovary cells in vitro in both the presence and absence of S9 mix.  It was tested to the maximum permitted concentration of 5000 µg.ml-1 in the absence of S9 mix and up to 80 µg.ml-1 in the presence of S9 mix (Test 1).  Toxicity was noted at 60 and 80 µg.ml-1 in the presence of S9 mix and at 156-5000 µg.ml-1 in the absence of S9 mix.  In Test 2, Tall Oil, CAS No. 8002-26-4, was tested up to 60 µg.ml-1 in the presence of S9 mix and up to 70 µg.ml-1 in the absence of S9 mix. Toxicity was noted in cultures treated with 30-60 µg.ml-1 (presence of S9 mix) and in cultures treated with 70 µg.ml-1 (absence of S9 mix).

It was concluded that Tall Oil, CAS No. 8002-26-4, was weakly clastogenic when tested with Chinese hamster ovary cell in vitro in the presence of S9 mix.  The response was observed only at a concentration level deemed overtly toxic to the cells.