Glycerol trioctanoate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

Acceptable, well documented study report which meets basic scientific principles.

Data source

Reference

Reference 1

Reference Type:

publication

Title:

Unnamed

Year:

1989

Reference 2

Reference Type:

publication

Title:

Prediction of Salmonella mutagenicity

Author:

Zeiger, E. et al.

Year:

1996

Bibliographic source:

Mutagenesis Vol. 11, No. 5: 471-484

Reference 3

Reference Type:

publication

Title:

Unnamed

Year:

1994

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted in 1981

Deviations:

yes

Remarks:

S9 mix from hamster used

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted in 1997

Deviations:

yes

Remarks:

2-Aminoanthracene was used as the sole indicator of the efficacy of the S9-mix; only four strains were tested and no strain was included to cross-linking mutagens (e.g. TA102 or E.coli); lack of cytotoxicity data.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Glycerol trioctanoate

EC Number:

208-686-5

EC Name:

Glycerol trioctanoate

Cas Number:

538-23-8

Molecular formula:

C27H50O6

IUPAC Name:

1,3-bis(octanoyloxy)propan-2-yl octanoate

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 97, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats and hamsters treated with Aroclor 1254

Test concentrations with justification for top dose:

TA100 / TA97/ TA 98: 100, 333, 1000, 3333, 10000 µg/plate with and without S9 mix
TA 1535: 100, 333, 1000, 3333, 6666, 10000, 16666 µg/plate with and without S9 mix

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: -S9: 2-nitrofluorene (TA98), sodium azide (TA100 and TA1535), 9-aminoacridine (TA97); +S9: 2-aminoanthracene or occasionally sterigmatocystin (all strains)

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation, agar plates

DURATION
- Preincubation period: 20 min. at 37°C
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: triplicates

DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency

Evaluation criteria:

If the test chemical was mutagenic to any particular strain of bacterium, the number of histidine-independent colonies arising on those plates must be significantly greater than the corresponding control plates for that strain of bacteria. The positive control plates were also counted, and the number of mutant colonies appearing on them must be significantly increased over the spontaneous control number for the test to be considered valid. Failure of the positive control chemical to induce mutation is reason to discard the experiment.
In analyzing the data, the pattern and the strength of the mutant response are taken into account in determining the mutagenicity of a chemical. If no increase in mutant colonies is seen after testing several strains under several different culture conditions, the test chemical is considered to be nonmutagenic in the Ames test.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium, other: TA 97, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Remarks:

but tested up to limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

at concentrations >= 6666 µg/plate

Cytotoxicity / choice of top concentrations:

not specified

Remarks:

but tested up to limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Remarks:

but tested up to limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other:

Remarks:

in the presence of hamster or rat S9 mix

Any other information on results incl. tables

Table 1: Mutant frequency (revertants per plate ± standard error from 3 plates) in TA1535, TA97, TA98, TA100 after treatment with the test substance.

Strain	Dose (µg/plate)	-S9		+30% Hamster S9						+30% rat S9
				Trial 1		Trial 2		Trial 3		Trial 1		Trial 2		Trial 3
		Mean	± SEM	Mean	± SEM	-	-	-	-	Mean	± SEM	-	-	-	-
TA100	0	144	17.7	137	7	-	-	-	-	193	8.5	-	-	-	-
	100	169	5.2	135	4.9	-	-	-	-	197	4.8	-	-	-	-
	333	134	17	142	6.2	-	-	-	-	189	6.5	-	-	-	-
	1000	108	1.9	147	5.7	-	-	-	-	190	3.8	-	-	-	-
	3333	112	1	137	5	-	-	-	-	176	4.5	-	-	-	-
	10000	131	5.2	176	2.2	-	-	-	-	188	9.5	-	-	-	-
	Positive Control*	955	37.8	837	39.5	-	-	-	-	440	26.1	-	-	-	-
Strain	Dose (µg/plate)	-S9		+30% Hamster S9						+30% rat S9
				Trial 1		Trial 2		Trial 3		Trial 1		Trial 2		Trial 3
		Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM
TA1535	0	12	0.9	15	0.9	11	0	13	2.6	18	3.8	15	1.2	20	3.8
	100	14	0.9	11	2.3	-	-	-	-	16	3.8	-	-	-	-
	333	13	3.4	14	2	-	-	-	-	17	1.2	-	-	-	-
	1000	15	2.6	12	0.9	12	2.9	24	1.2	17	2.3	15	1.5	28	5.1
	3333	14	1.3	15	0.3	18	1.5	45	4.5	20	2.6	19	1.7	37	1.5
	6666	-	-	-	-	50	6	42	1.9	-	-	37	2.2	39	3.4
	10000	17	1.8	44	9.1	52	2.8	55	1.9	42	1.5	49	5.5	54	3.8
	16666	-	-	-	-	85	2.8	107	10.7	-	-	76	2.4	95	3.2
	Positive Control	958	34.3	607	16.8	408	39.8	351	8.9	105	3.6	91	9.1	90	10.5
Strain	Dose (µg/plate)	-S9		+30% Hamster S9						+30% rat S9
				Trial 1		Trial 2		Trial 3		Trial 1		Trial 2		Trial 3
		Mean	± SEM	Mean	± SEM	-	-	-	-	Mean	± SEM	-	-	-	-
TA97	0	222	6.9	185	6.9	-	-	-	-	216	5.3	-	-	-	-
	100	228	3.8	198	7.3	-	-	-	-	205	20.4	-	-	-	-
	333	223	6.3	210	7.6	-	-	-	-	224	11.9	-	-	-	-
	1000	210	2.6	216	1.8	-	-	-	-	184	6.5	-	-	-	-
	3333	212	7.6	203	21.5	-	-	-	-	168	3.7	-	-	-	-
	10000	225	4	197	19.4	-	-	-	-	152	10.4	-	-	-	-
	Positive Control	742	42.2	436	2.2	-	-	-	-	442	3.5	-	-	-	-
Strain	Dose (µg/plate)	-S9		+30% Hamster S9						+30% rat S9
				Trial 1		Trial 2		Trial 3		Trial 1		Trial 2		Trial 3
		Mean	± SEM	Mean	± SEM	-	-	-	-	Mean	± SEM	-	-	-	-
TA98	0	30	4.7	35	4	-	-	-	-	38	1.9	-	-	-	-
	100	27	1.5	34	2.4	-	-	-	-	41	0.6	-	-	-	-
	333	27	0.9	33	6.7	-	-	-	-	36	3.8	-	-	-	-
	1000	28	4.6	29	1.7	-	-	-	-	31	3.7	-	-	-	-
	3333	28	1.5	28	3.4	-	-	-	-	34	1.7	-	-	-	-
	10000	28	2	31	4.3	-	-	-	-	28	1.5	-	-	-	-
	Positive Control	677	20.6	770	11.3	-	-	-	-	168	3.5	-	-	-	-

* The positive controls in the absence of metabolic activation were sodium azide (TA100 and TA1535), 9-aminoacridine (TA97), and 4-nitro-o-phenylenediamine (TA98). The positive control for metabolic activation with all strains was 2-aminoanthracene.

Applicant's summary and conclusion

Conclusions:

Negative in TA 97, TA98 and TA 100 with and without metabolic activation (hamster S9 mix or rat S9 mix).
Positive with metabolic activation in TA1535, but only at very high concentrations of 6666 to 16666 µg/plate, which is above the required limit concentration of 5000 µg/plate.