Reaction mass of 3-(octanoyloxy)propyl decanoate and propane-1,3-diyl didecanoate and propane-1,3-diyl dioctanoate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30.06.1995 - 17.07.1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted May 1983

Principles of method if other than guideline:

Only 4 bacterial strains used.

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Decanoic acid, mixed diesters with octanoic acid and propylene glycol

EC Number:

271-516-3

EC Name:

Decanoic acid, mixed diesters with octanoic acid and propylene glycol

Cas Number:

68583-51-7

Molecular formula:

C21H44O6

IUPAC Name:

Decanoic acid, mixed diesters with octanoic acid and propylene glycol

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

Type and composition of metabolic activation system:
- source of S9 : CCR (Cytotest Cell Research GmbH & Co. KG), Roßdorf, Germany
- method of preparation of S9 mix: according to Ames et al., 1975 (obtained from the livers of male Wistar rats which received a single i.p. injection of Aroclor 1254 (500 mg/kg body weight) in olive oil 5 days prior to the preparation of the S9 fraction. The livers were removed from the animals, homogenized, and the supernatant of the 9000 x g centrifugation step (the "59 fraction") was frozen immediately. Protein content, sterility and activity of the preparation in the S. typhimurium gene mutation assay were certified by CCR.)

Test concentrations with justification for top dose:

50, 160, 500, 1600 and 5000 µg/plate

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-aminoanthracene (with S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 h at 37°C

SELECTION AGENT: - Salmonella typhimurium: L-Histidine

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: reduced growth of the bacterial background lawn

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

A reduced growth of the bacterial background lawn, indicative of test compound induced cytotoxicity, was not detectable with any of the tester strains. In both tests with and without metabolic activation by S9 mix, the test item formed a precipitate at the highest concentration tested (5000 µg/plate). The precipitate did not interfere with colony counting and obviously did not effect the mutant frequency of any of the S. typhimurium strains. All four bacterial strains exhibited a positive mutagenic response with the positive controls tested both with and without metabolic activation by S9 mix. Negative (solvent) controls were also tested with each strain, and the mean numbers of spontaneous revertants were considered acceptable. In both experiments, no indication of test compound induced mutagenicity was observed with either one of the four tester strains TA 98, TA 100, TA 1535, and TA 1537, with or without metabolic activation.

Any other information on results incl. tables

Table 1: Mean number of revertants with S9 mix

	Dose/Plate (µg)	TA 1535	TA1537	TA 98	TA 100
Deionised water	-	13	16	42	125
acetone	-	9	19	44	125
Test substance	50	13	19	47	130
	160	11	15	57	120
	500	9	15	60	117
	1600	9	15	59	117
	5000	10	18	64	125
Positive control 1	2.5	197	113	1396	1617

Positive Control 1: 2 –Aminoanthracene

  

Table 2: Mean number of revertants without S9 mix

	Dose/Plate (µg)	TA 1535	TA1537	TA 98	TA 100
Deionised water	-	8	11	20	108
acetone		9	11	22	98
Test substance	50	5	17	17	108
	160	7	16	20	122
	500	7	16	21	105
	1600	5	15	23	89
	5000	6	15	20	113
Positive control 1	2.5			115
Positive control 2	2.5	330			372
Positive control 3	25		47

Positive control 1:2-Nitronuorene

Positive control 2:Sodium azide

Positive control 3:9-Aminoacridine

Applicant's summary and conclusion

Conclusions:

The mutagenicity of the test item was determined in a study according to OECD guideline 471. The results indicate that the test item under the experimental conditions described, was not mutagenic to Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 in the presence and absence of a metabolizing system.

Executive summary:

The mutagenicity of the test substance was studied with the mutant strains of Salmonella typhimurium (TA1535, TA1537, TA98 and TA100) according to OECD guideline 471. The investigations were carried using the standard plate incorporation assay with and without liver homogenate (S9) from Aroclor 1254 pretreated male rats as metabolic activation system. The test substance was dissolved in DMSO and tested in concentrations of 50 to 5000 µg per plate in the presence and absence of S9. Precipitation of the test compound an the plates was observed at 5000 µg/plate. All four bacterial strains exhibited a positive mutagenic response with the positive controls tested both with and without metabolic activation by S9 mix. Negative (solvent) controls were also tested with each strain, and the mean numbers of spontaneous revertants were considered acceptable. In the concentration range investigated, the test substance did not induce a significant increase in the mutation frequency of the tester strains in the presence and absence of a metabolic activation system. In conclusion, these results indicate that under the experimental conditions described, the test item was not mutagenic to Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 in the presence and absence of a metabolizing system.