1,2,3-propanetriyl triisooctadecanoate

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:

key study

Study period:

07 Mar-21 Mar 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study with acceptable restrictions. Only 4 S. typhimurium strains tested, no TA102 or E. coli strain was included.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

only 4 S. typhimurium strains tested, no TA102 or E. coli strain included

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

yes

Remarks:

only 4 S. typhimurium strains tested, no TA102 or E. coli strain included

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,2,3-propanetriyl triisooctadecanoate

EC Number:

248-122-5

EC Name:

1,2,3-propanetriyl triisooctadecanoate

Cas Number:

26942-95-0

Molecular formula:

C57H110O6

IUPAC Name:

1,2,3-Propanetriyl tris(16-methylheptadecanoate)

Details on test material:

- Name of test material (as cited in study report): only trade name given
- Physical state: yellow liquid
- Analytical purity: no data
- Lot/batch No.: 8779
- Storage condition of test material: at room temperature in the dark

Method

Target gene:

his operon

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:

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

Test concentrations with justification for top dose:

Preliminary experiment: 3, 10, 33, 100, 333, 1000, 3330 and 5000 µg/plate with and without metabolic activation
First experiment: 100, 333, 1000, 3330 and 5000 µg/plate with and without metabolic activation
Second experiment: 100, 333, 1000, 3330 and 5000 µg/plate with and without metabolic activation

Vehicle / solvent:

- Vehicle used: DMSO (0.1 mL per plate)

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: without S9: 9-aminoacridine (9AC; 60 µg/plate in saline, TA1537); sodium azide (SA; 1 µg/plate in saline, TA1535); daunomycine (DM; 4 µg/plate in saline, TA98); methylmethanesulfonate (MMS; 650 µg/plate in DMSO, TA100)

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: with S9: 2-amino-anthracene (2-AA; 0.5 or 5 µg/plate in DMSO for all strains)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: triplicates each in two independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: Inspection of bacterial background lawn

Evaluation criteria:

A test substance is considered positive (mutagenic) in the test if:
a) It induces at least a 2-fold, dose related increase in the number of revertants with respect to the number induced by the solvent control in any of the tester strains, either with of without metabolic activation. However, any mean plate count of less than 20 revertants is considered to be not significant.
b) The positive response should be reproducible in at least one independently repeated experiment.

Statistics:

Mean values and standard deviation were calculated.

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: yes, at concentrations of 5000 µg/plate.

RANGE-FINDING/SCREENING STUDIES:
Strain TA100 was used for testing up to 5000 µg/plate with and without S9-mix. The results of the dose range finding experiment were included into the first experiment.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
No reduction of the bacterial background lawn and no decrease in the number of revertants was observed. Therefore, the test substance was tested up to a concentration of 5000 µg/plate in the absence or presence of S9-mix.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1. Ames Test Results of Experiment 1.

With or without S9-Mix	Test substance concentration	Mean number of revertant colonies per plate
	(μg/plate)	(average of 3 plates ± Standard deviation)
		Base-pair substitution type		Frameshift type
		TA 100	TA1535	TA98	TA1537
–	0	63 ± 6	15 ± 2	13 ± 1	7 ± 3
–	3	59 ± 1
–	10	70 ± 6
–	33	73 ± 11
–	100	65 ± 1	13 ± 5	15 ± 5	7 ± 3
–	333	57 ± 8	15 ± 3	18 ± 4	7 ± 1
–	1000	53 ± 12	15 ± 4	15 ± 3	9 ± 2
–	3330	54 ± 2	12 ± 2	16 ± 4	8 ± 1
–	5000*	54 ± 5	15 ± 3	17 ± 5	7 ± 4
Positive controls, –S9	Name	MMS	SA	DM	9AC
	Concentrations (μg/plate)	650	1	4	60
	Mean No. of colonies/plate (average of 3 ± SD)	804 ± 25	6142 ± 5	241; 186**	238 ± 113
+	0	76 ± 7	13 ± 6	18 ± 6	7 ± 1
+	3	66 ± 18
+	10	71 ± 6
+	33	67 ± 11
+	100	54 ± 9	17 ± 1	18 ± 7	8 ± 2
+	333	63 ± 13	11 ± 2	22 ± 2	4 ± 1
+	1000	59 ± 16	15 ± 2	23 ± 1	6 ± 4
+	3330	66 ± 15	12 ± 6	20 ± 4	6 ± 3
+	5000*	58 ± 6	9 ± 3	18 ± 6	7 ± 2
Positive controls, +S9	Name	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	0.5	5	0.5	5
	Mean No. of colonies/plate (average of 3 ± SD)	647 ± 69	246 ± 14	471 ± 20	572 ± 39

* Test subtance precipitated slightly on the plates

**One plate infected with other bacteria

SA = Sodium azide

9AC = 9 Aminoacridine

DM = Daunomycine

MMS = Methylmethanesulfonate

2AA = 2 -Aminoanthracene

Table 2. Ames Test Results of Experiment 2.

With or without S9-Mix	Test substance concentration	Mean number of revertant colonies per plate
	(μg/plate)	(average of 3 plates ± Standard deviation)
		Base-pair substitution type		Frameshift type
		TA 100	TA1535	TA98	TA1537
–	0	60 ± 1	6 ± 3	14 ± 3	4 ± 1
–	100	56 ± 8	7 ± 2	15 ± 6	5 ± 1
–	333	68± 3	9 ± 2	14 ± 5	5 ± 1
–	1000	68 ± 9	6 ± 1	13 ± 1	7 ± 1
–	3330	71 ± 6	9 ± 3	15 ± 3	6 ± 2
–	5000*	58 ± 10	11 ± 3	14 ± 1	9 ± 3
Positive controls, –S9	Name	MMS	SA	DM	9AC
	Concentrations (μg/plate)	650	1	4	60
	Mean No. of colonies/plate (average of 3 ± SD)	1047 ± +65	178 ± 9	194 ± 11	204 ± 43
+	0	73 ± 3	10 ± 3	20 ± 2	7 ± 3
+	100	70 ± 15	6 ± 2	17 ± 4	5 ± 1
+	333	72 ± 13	11 ± 1	22 ± 9	6 ± 2
+	1000	95 ± 9	13 ± 2	24 ± 2	8 ± 1
+	3330	89 ± 15	7 ± 1	23 ± 6	7 ± 1
+	5000*	81 ± 4	9 ± 4	19 ± 3	7 ± 2
+	Name	2AA	2AA	2AA	2AA
+	Concentrations (μg/plate)	0.5	5	0.5	5
+
Positive controls, +S9	Mean No. of colonies/plate (average of 3 ± SD)	1169 ± 87	475 ± 12	593 ± 6	669 ± 93

SA = Sodium azide

9AC = 9 -Aminoacridine

DM = Daunomycine

MMS = Methylmethanesulfonate

2AA = 2 -Aminoanthracene

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative