Reaction mass of glycerol, glycerol 1-formate, glycerol 2-formate, glycerol 1,2-diformate, glycerol 1,3-diformate and glycerol triformate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

This endpoint study record is part of a Weight of Evidence approach comprising of read-across from three analogue source substance studies. The results of the read-across studies agree as to the potential for cytogenicity in mammalian cells and are sufficient to fulfil the information requirements as further explained in the provided genetic toxicity endpoint summary.

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1984

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosomal Aberration Test)

Deviations:

yes

Remarks:

no metabolic activation used

GLP compliance:

not specified

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

Glycerol

EC Number:

200-289-5

EC Name:

Glycerol

Cas Number:

56-81-5

Molecular formula:

C3H8O3

IUPAC Name:

Propane-1,2,3-triol

Details on test material:

GLYCERINE USP grade

Specific details on test material used for the study:

Reported as: Glycerin
Purity: 99.4%

Method

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

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, 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.

Metabolic activation:

without

Test concentrations with justification for top dose:

up to 1.0 mg/mL

Vehicle / solvent:

Physiological saline

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Test involved screening a large number of substances, some of which (e.g., hydrogen peroxide, sodium nitrite) gave positive results but no specific positive control as recommended in the guideline was used.

Details on test system and experimental conditions:

METHOD OF APPLICATION: Colcemid (final concentration 0.2 µg/ml) was added to the culture 2 hr before cell harvesting, trypsinized and suspended in a hypotonic KCI solution (0.075 M) for 13 min at room temperature. The cells were centrifuged, fixed with acetic acid-methanol (I :3, v/v) and spread on clean glass slides, air-dried, and stained with Giemsa solution (1.5%, at pH 6.8) for 12-15 min.

DURATION
- Exposure duration: 24 and 48 h

NUMBER OF CELLS EVALUATED: A hundred well-spread metaphases were observed under the microscope (x 600 with a no-cover objective lens).

OTHER EXAMINATIONS:
- Other: 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.

OTHER: The cells were exposed to each sample at three different doses

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%. For a quantitative evaluation of the clastogenic potential, the dose at which structural aberrations (including gaps) were detected in 20% of the metaphases was calculated (D20) along with the frequency of cells with exchange type aberrations per unit dose (mg/mL - TR value). Low TR values indicate low carcinogenic potential in man.

Results and discussion

Test results

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Treatment of Chinese hamster lung fibroblasts with SS1 at doses of up to 1.0 mg/mL for a period of either 24- or 48-hours was reported to not increase the frequencies of chromosome aberrations. In the 48-hour exposure experiment, the incidence of structural aberrations and polyploidy was 1.0 and 2.0%, respectively.

Applicant's summary and conclusion

Conclusions:

The authors concluded that the substance was not genotoxic in the in vitro mammalian chromosome aberration assay.