Fatty acids, vegetable-oil, esters with dipropylene glycol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

26 Mar - 05 Apr 1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP - Guideline study with acceptable restrictions. No S. typhimurium TA102 or E. coli strain was included.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

no S. typhimurium TA102 or E. coli strain was included

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

no S. typhimurium TA102 or E. coli strain was included

GLP compliance:

yes

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

IUPAC Name:

68583-51-7

Details on test material:

- Name of test material (as cited in study report): [trade name], Decanoic acid, mixed diesters with octanoic acid and propylene glycol
- Analytical purity: 100%
- Physical state: colourless liquid
- Lot/batch No.: 3409
- Production date: November 04, 1990
- Expiration date of the lot/batch: May 04, 1992
- Storage condition of test material: at room temperature
- Stability:
Pure: 18 months
In Solvent: not given
- Receipt of the test substance: February 08, 1991

Method

Target gene:

his operon

Species / strain

Species / strain 1

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

- Type and identity of media: Standard I nutrient broth for microbiology

Additional strain / cell type characteristics:

other: rfa-; uvrB- (R+ for TA98 and TA100)

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Details on mammalian cell type (if applicable):

- Type and identity of media: Standard I nutrient broth for microbiology

Additional strain / cell type characteristics:

other: rfa-; uvrB-

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:

First and second experiment: 8, 40, 200, 1000 and 5000 µg/plate with and without metabolic activation

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Suspension medium Tween 80/bidest. water
- Justification for choice of solvent/vehicle: According to the author, the suspension medium was chosen based on the solubility properties preliminary tested before start of the study.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

Tween 80/bidest. water

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: sodium azide (2 µg/plate, ±S9, TA1535 and TA100); 9-Aminoacridine (80 µg/plate, ±S9, TA1537); 4-Nitro-o-phenylendiamine (40 µg/plate, ±S9,TA98 and TA1538); 2 Amino-anthracene (2.5 or 5 µg/plate, ±S9, TA1535 and TA1357; TA100, TA1538 and TA98)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 replications each in 2 independent experiments

Evaluation criteria:

The test material may be considered positive in this test system if a combination of the following criteria are met:
- the plate background of non-reverted bacteria does not show any growth reduction versus the respective negative controls.
- the spontaneous mutation rates of each tester strain per plate are within the characteristic spontaneous mutation range.
- the positive controls show mutation rates exceeding the control values of TA100 at least two fold and those of the other strains at least by the factor 3.
- at more than one dose tested, at least a two-fold (or more) increase in comparison with the negative controls in the tester strain TA100. For the other strains, an increase in the mutation rate of more than 3 above the corresponding negative controls.

Statistics:

Mean values and standard deviation were calculated.

Results and discussion

Test results

Test results 1

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

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and 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:

COMPARISON WITH HISTORICAL CONTROL DATA:
- see Table 1 and Table 2

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

COMPARISON WITH HISTORICAL DATA

Table 1. Characteristic spontaneous mutation range of the test batches without S9-mix.

Tester strains	TA1535	TA100	TA1537	TA1538	TA98
Historical laboratory values (Mean values) Extreme values	6 1-25	87 35-201	7 1-24	15 6-27	20 5-39
Ames et al.* (Mean values) Extreme values	20 5-50	160 60-220	7 3-25	25 5-40	40 15-75

 *Ames, BN, McCann, J, Yamasaki, E 1977, ´Methods for detecting carcinogens and mutagens with the Salmonella/mammalian microsome mutagenicity test.`in Kilbey et al. Handbook of Mutagenicity Test Procedures, Elsevier, Amsterdam, pp. 1 -17.

Table 2. Characteristic spontaneous mutation range of the test batches containing S9-mix.

Tester strains	TA1535	TA100	TA1537	TA1538	TA98
Historical laboratory values (Mean values) Extreme values	8 1-25	105 54-252	6 1-23	18 3-48	27 7-76

STUDY RESULTS

Table 3. Test results of experiment 1 (plate incorporation).

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type		Frameshift type
		TA1535	TA100	TA1537	TA1538	TA98
–	Negative control	11.7 ± 2.3	99.7 ± 11.7	6.7 ± 3.8	8.7 ± 0.6	22.7 ± 1.5
–	Solvent control	12.7 ± 5.5	113.3 ± 6.4	6.3 ± 1.5	11.30 ± 2.1	27.0 ± 3.5
–	8	13.7 ± 1.5	103.3 ± 3.1	8.0 ± 3.6PM	7.7 ± 1.5	30.0 ± 5.3
–	40	9.3 ± 2.1	104.7 ± 12.0	6.3 ± 1.5	10.7 ± 3.8	26.3 ± 6.7
–	200	8.7 ± 3.1	113.0 ± 7.0	8.0 ± 2.0	11.0 ± 2.0	26.0 ± 2.7
–	1000	13.7 ± 4.2	104.7 ± 7.1	8.0 ± 3.0	13.3 ± 4.0	27.3 ± 3.5
–	5000	9.7 ± 0.6	110.0 ± 8.9	7.0 ± 3.6	7.3 ± 2.3	24.3 ± 2.1
Positive controls, –S9	Name	SA	SA	9AA	4NPD	4NPD
	Concentrations (μg/plate)	2	2	80	40	40
	Mean No. of colonies/plate (average of 3 ± SD)	534.7 ± 38.4	324.0 ± 19.2	352.0 ± 57.1	1428.0 ± 19.0	699.0 ± 99.0
	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	2.5	5	2.5	5	5
	Mean No. of colonies/plate (average of 3 ± SD)	9.7 ± 0.6	110.7 ± 7.6	10.7 ± 2.1	18.7 ± 6.1	27.3 ± 1.2
+	Negative control	12.7 ± 3.1	101.7 ± 7.8	9.3 ± 2.1	18.7 ±3.2	42.3 ± 5.0
+	Solvent control	14.7 ± 3.5	107.0 ± 15.4	9.3 ± 3.2	15.3 ± 2.1	45.0 ± 5.0
+	8	14.0 ± 4.4	103.7 ± 23.0	8.0 ± 1.7	12.7 ± 2.1	42.7 ± 4.7
+	40	14.3 ± 2.1	104.3 ± 12.7	6.3 ± 1.2	15.7 ± 2.9	49.3 ± 2.1
+	200	11.7 ± 1.5	115.0 ± 10.4	7.0 ± 1.0	14.7 ± 4.7	38.7 ± 2.3
+	1000	16.0 ± 2.0	116.7 ± 11.9	5.3 ± 1.5	17.0 ± 2.0	45.3 ± 8.1
+	5000	6.3 ± 0.6	81.0 ± 4.6	6.3 ± 3.2	16.0 ± 2.7	31.3 ± 0.6
Positive controls, +S9	Name	SA	SA	9AA	4NPD	4NPD
	Concentrations (μg/plate)	2	2	80	40	40
	Mean No. of colonies/plate (average of 3 ± SD)	143.0 ± 17.8	180.3 ± 8.1	268.3 ± 18.5	1170.3 ± 84.0	617.3 ± 169.5
	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	2.5	5	2.5	5	5
	Mean No. of colonies/plate (average of 3 ± SD)	181.0 ± 19.3	1059.3 ± 129.7	95.7 ± 16.3	1219.7 ± 152.3	875.0 ± 40.5

SA = sodium azide

4NPD = 4-nitro-o-phenylendiamine

9AA = 9-aminoacridine

2AA = 2-Amino-anthracene

PM = in part manual evaluation

Table 4. Test results of experiment 2 (plate incorporation). 

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type		Frameshift type
		TA1535	TA100	TA1537	TA1538	TA98
–	Negative control	9.7 ± 1.5	88.3 ± 10.8	10.3 ± 4.0	9.7 ± 2.5	28.7 ± 2.5
–	Solvent control	8.3 ± 1.5	86.7 ± 19.7	8.7 ± 1.2	10.7 ± 1.2	26.3 ± 4.0
–	8	9.3 ± 1.5	86.0 ± 3.5	7.3 ± 0.6	9.0 ± 1.0	24.7 ± 4.0
–	40	10.3 ± 0.6	91.7 ± 18.3	8.7 ± 2.1	9.3 ± 1.5	27.0 ± 6.6
–	200	6.7 ± 2.5	92.7 ± 4.5	9.3 ± 3.1	8.0 ± 2.7	20.7 ± 1.5
–	1000	10.3 ± 3.1	92.7 ± 5.9	8.0 ± 4.6	8.7 ± 5.5	31.3 ± 1.2
–	5000	7.7 ± 3.1PM	92.3 ± 20.0	9.0 ± 4.0	8.7 ± 1.2	28.3 ± 5.1
Positive controls, –S9	Name	SA	SA	9AA	4NPD	4NPD
	Concentrations (μg/plate)	2	2	80	40	40
	Mean No. of colonies/plate (average of 3 ± SD)	357.3 ± 26.8	216.7 ± 15.3	692.0 ± 186.2	1242.3 ± 56.2	535.0 ± 49.7
	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	2.5	5	2.5	5	5
	Mean No. of colonies/plate (average of 3 ± SD)	9.3 ± 2.1	80.3 ± 8.2	6.7 ± 2.9	14.3 ± 2.5	32.3 ± 4.6
+	Negative control	7.0 ± 2.7	72.3 ± 8.0	7.0 ± 1.0	12.7 ± 0.6M	30.0 ± 7.6M
+	Solvent control	9.7 ± 2.3	80.0 ± 12.1	7.0 ± 1.0	10.7 ± 0.6M	37.3 ± 4.7M
+	8	6.7 ± 1.2	64.7 ± 5.5	8.7 ± 1.5M	14.3 ± 1.5M	36.3 ± 6.7M
+	40	5.3 ± 0.6	72.3 ± 9.7	7.3 ± 1.2M	11.7 ± 1.5M	31.7 ± 3.2M
+	200	8.7 ± 1.5	77.0 ± 14.8	7.7 ± 2.1M	11.3 ± 2.5M	30.7 ± 9.5M
+	1000	7.7 ± 1.5	69.7 ± 4.0	6.0 ± 1.0M	11.0 ± 1.0M	30.7 ± 8.5M
+	5000	5.0 ± 1.0	58.0 ± 2.0	3.3 ± 1.5M	6.3 ± 0.6M	22.7 ± 4.0M
Positive controls, +S9	Name	SA	SA	9AA	4NPD	4NPD
	Concentrations (μg/plate)	2	2	80	40	40
	Mean No. of colonies/plate (average of 3 ± SD)	57.0 ± 3.6	98.0 ± 11.4	428.3 ± 29.3	986.0 ± 89.9M	720.0 ± 16.0M
	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	2.5	5	2.5	5	5
	Mean No. of colonies/plate (average of 3 ± SD)	90.3 ± 20.8	467.0 ± 22.5	105.3 ± 12.1	1072.0 ±57.7M	909.3 ± 159.8M

SA = sodium azide

4NPD = 4-nitro-o-phenylendiamine

9AA = 9-aminoacridine

2AA = 2-Amino-anthracene

M = manual evaluation

PM = in part manual evaluation

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative