coconut oil, reaction products with glycerol esters of 3,5-bis(1,1-dimethylethyl)-4-hydroxybenzenepropanoic acid

Genetic toxicity: in vitro

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: Guideline study (OECD, GLP, QAU)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Version / remarks:

adopted May 26, 1983

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

425-400-6

EC Name:

-

Cas Number:

179986-09-5

Molecular formula:

UVCB substance

IUPAC Name:

methyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl)propanoate; propane-1,2,3-triol; tetradecanoic acid

Details on test material:

- Physical state: liquid
- Analytical purity: 95%
- Storage condition of test material: room temperature

Method

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Metabolic activation system:

liver S9-mix from Phenobarbital/p-Naphthoflavone treated rats

Test concentrations with justification for top dose:

experiment I: 30-5000 µg/ml (with metabolic activation)
experiment I: 50-300 µg/ml (without metabolic activation)
experiment II: 30-5000 µg/ml (with metabolic activation)
experiment II: 50-200 µg/ml (without metabolic activation)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: solubilty and its non-toxicity to the cells

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: cyclophosphamide (2.5 µM, with metabolic activation); ethylmethanesulphonate (4.8 mM, without metabolic avtivation)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 4 hours with metabolic activation, 18 or 28 hours without metabolic activation

SPINDLE INHIBITOR (cytogenetic assays): colcemide (treatment of 2.5 h)
STAIN (for cytogenetic assays): Giemsa

NUMBER OF METAPHASES EVALUATED: 100 well spread metaphases per culture were scored for cytogenetic damage on coded slides.

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index

OTHER EXAMINATIONS:
- Determination of polyploidy: yes

Evaluation criteria:

A test article is classified mutagenic if it induces reproducibly either a significant concentration-related increase in the number of structural chromosome aberrations or a significant and reproducible positive response for at least one of the test concentrations. A test article producing reproducibly neither a significant concentration-related increase in the number of structural chromosome aberrations nor a significant and reproducibly positive response at any one of the test points is considered non-mutagenic in this system.

Statistics:

Fischer's exact test

Results and discussion

Test results

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

ambiguous

Remarks:

some aberrations were noted but no statistically significant increases in the frequency of aberrations per cell or in the proportion of aberrant metaphases at any dose level evaluated with or without S9 Mix were observed.

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

without metabolic activation: mitotic indices were reduced to values below 60 % of the corresponding control in the evaluated cultures (at interval 18 and 28 h); with metabolic activation: mitotic index was slightly reduced at interval 28 h (about 30 %)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

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 chromosomal aberration in V79 cells after treatement with test substance

Experiment	Preparation interval (hours)	S9 Mix	Concentration in µg/ml	Aberrant cells (incl./excl. gaps, exchanges, %)
I	18	-	solvent control	6.5
		-	50	16
		-	100	20
		-	300^P	3
II	18	-	solvent control	5.5
		-	50	4.5
		-	100	8
		-	200	14
I	28	-	solvent control	5.5
		-	300^P	9.5
II	28	-	solvent control	5
		-	100	4
I	18	+	solvent control	5.5
		+	30	7.5
		+	100	5
		+	300^P	11.5
		+	5000^P	15.5
II	18	+	solvent control	3
		+	30	4.5
		+	100	2.5
		+	300^P	5.5
		+	5000^P	5.5
I	28	+	solvent control	3
		+	100	13.5
		+	5000^P	5.5
II	28	+	solvent control	4.5
		+	100	5
		+	5000^P	4

p: visible precipitation was observed