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Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

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Administrative data

in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Reason / purpose for cross-reference:
reference to same study

Data source

Reference Type:
Primary mutagenicity screening of food additives currently used in Japan
M. Ishidate Jr, T. Sofuni, K. Yoshikawa, M. Hayashi, T. Nohmi, M. Sawada and A. Matsuoka
Bibliographic source:
Fd Chem. Toxic. Vol. 22, No. 8, pp. 623-636

Materials and methods

Test guidelineopen allclose all
according to guideline
other: method of Ishidate & Odashima (1977)
equivalent or similar to guideline
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
GLP compliance:
not specified
Type of assay:
other: in vitro mammalian chromosome aberration test

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
EC Name:
Cas Number:
Molecular formula:


Species / strain
Species / strain / cell type:
other: Chinese Hamster Lung Fibroblast (CHL)
Details on mammalian cell type (if applicable):
The cell line was originally established from the lung of a newborn female at the Cancer Research Institute, Tokyo (Koyama, Utakoji & Ono, 1970), and was maintained by 4-day passages in Minimum Essential Medium (MEM; GIBCO) supplemented by 10% calf serum.
The modal chromosome number is 25 and the doubling time was approximately 15 hr.
Additional strain / cell type characteristics:
not specified
Metabolic activation:
Test concentrations with justification for top dose:
3 different concentrations tested but not specified in the publication.
The maximum dose of each sample was selected by a preliminary test in which the dose needed for 50% cell-growth inhibition was estimated using a cell densitometer (Monocellater, Olympus Co., Ltd).
Previous studies indicated that the osmotic pressure of the medium generally rose with sample concentrations of more than 10 mM, so that the maximum dose for some samples was limited to around this level, at which cytotoxic effects were not necessarily ubserved.
Vehicle / solvent:
No justification given
Untreated negative controls:
untreated cultures
Negative solvent / vehicle controls:
same treatment but exposed only to DMSO
True negative controls:
not specified
Positive controls:
not specified
Positive control substance:
not specified
Details on test system and experimental conditions:
The cells were exposed at three different doses for 24 and 48 hr. In the present study, no metabolic activation systems were applied.

Chromosome preparations were made as follows:
Colcemid (final concn 0.2 µg/ml) was added to the culture 2 hr before cell harvesting. The cells were then trypsinized and suspended in a hypotonic KCI solution (0.075 M) for 13 min at room temperature. After centrifugation the cells were fixed with acetic acid-methanol (1:3, vJv) and spread on clean glass slides. After air-drying, the slides were stained with Giemsa solution (1.5%, at pH 6.8; E. Merck) for 12-15 min. A hundred well-spread metaphases were observed under the microscope (x 600 with a no-cover objective lens). The incidence of polyploid cells as well as of cells with structural chromosomal aberrations such as chromatid or chromosome gaps, breaks, exchanges, ring formations, fragmentations and others, was recorded on each culture plate.

Untreated cells and solvent-treated cells served as negative controls, in which the incidence of aberrations was usually less than 3.0%.
Evaluation criteria:
The results were considered to be negative if the incidence was less than 4.9%, equivocal if it was between 5.0 and 9.9%, and positive if it was more than 10.0%.
no documentation available

Results and discussion

Test results
Species / strain:
other: Chinese Hamster Lung Fibroblast (CHL)
Metabolic activation:
at the highest non cytotoxic dose tested (0.2 mg/ml)
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
not specified
Untreated negative controls validity:
Positive controls validity:
not specified
Additional information on results:
Percentage of structure aberrations after 48 hours: 4%
no further details available

Any other information on results incl. tables

no detailed table given

Applicant's summary and conclusion

Interpretation of results: negative

The test material is not clastogenic under the used test conditions