Fatty acids, C8-14, triesters with trimethylolpropane

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 based on grouping of substances (category approach)

Adequacy of study:

weight of evidence

Study period:

31 Aug - 09 Sep 1999

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions S. typhimurium TA102 or E. coli WP2 were not tested

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted in 1983

Deviations:

yes

Remarks:

S. typhimurium TA102 or E. coli WP2 were not tested

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C8-18 and C18-unsatd., esters with trimethylolpropane

EC Number:

286-075-2

EC Name:

Fatty acids, C8-18 and C18-unsatd., esters with trimethylolpropane

Cas Number:

85186-89-6

IUPAC Name:

85186-89-6

Details on test material:

Name of test material (as cited in study report): Only trade name given
- Physical state: Liquid, yellow
- Analytical purity: 100%
- Lot/batch No.: 44538224
- Expiration date of the lot/batch: August 2000
- Stability under test conditions: not given
- Storage condition of test material: room temperature

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

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

Cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers treated with Phenobarbital (intraperitoneally) and beta-Naphthoflavone (orally)

Test concentrations with justification for top dose:

8, 40, 200, 1000, 5000 µg/plate (2 experiments, each with and without S9 mix)

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

acetone

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Other: -S9: sodium azide (2 µg/plate) for TA100 and TA1535; 9-aminoacridine (80µg/plate) for TA537; 4-nitro-o-phenylendiamine (40 µg/plate) for TA98 and TA1538; +S9: 2-aminoanthracene (2.5 or 5 µg/plate)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: duplicates

DETERMINATION OF CYTOTOXICITY
- Method: growth reduction of background bacteria was considered

Evaluation criteria:

The test material may be considered positive in this test system if the following criteria are met:
The plate background of non-converted bacteria did not show any growth reduction versus the respective negative controls.
The spontaneous mutation rates of each tester strain per plate were within the characteristic spontaneous mutation range.
The positive controls showed mutation rates exceeding the control values of TA 100 at least by the factor 2.0 and those of the other tester strains by the factor 3.0.
At more than one dose tested, the test substance caused at least a 2.0-fold increase in comparison with negative controls in the tester strain TA 100. For the other tester strains, and increase in the mutation rats of more than 3.0 above the corresponding negative controls was considered positive.

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 nor precipitates, but tested up to recommended limit concentrations

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 nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Table 1: Experiment 1:

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		TA 100	TA1535	TA1537	TA1538	TA98
-	Medium	113	11	11	8	20
-	Vehicle	78	8	7	6	18
-	8	78	8	5	6	16
-	40	80	8	4	6	18
-	200	80	10	5	6	28
-	1000	90	11	8	5	23
-	5000	90	6	5	5	22
Positive controls - S9	Name	SA	SA	9AA	4NP	4NP
	Concentrations (μg/plate)	2.0	2.0	80	40	40
	Number of colonies/plate	745	518	773	1875	1531
	Medium	119	16	8	13	21
+	Vehicle	91	11	6	7	18
+	8	118	17	9	6	21
+	40	110	10	6	10	18
+	200	109	14	5	9	23
+	1000	118	12	4	11	17
+	5000	110	10	9	8	23
Positive controls + S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	5.0	2.5	2.5	5.0	5.0
	Number of colonies/plate	318	264	257	2155	1808

SA: Sodium azide

9AA : 9-Aminoacridine

4NP : 4-Nitro-o-phenylenediamine

2-AA: 2-Aminoanthracene

 

 

Table 2: Experiment 2:

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		TA 100	TA1535	TA1537	TA1538	TA98
-	Medium	100	10	7	8	20
-	Vehicle	98	9	9	10	9
-	8	85	13	11	4	9
-	400	92	10	5	9	11
-	200	84	7	6	11	12
-	1000	77	13	6	8	13
-	5000	99	10	2	9	11
Positive controls - S9	Name	SA	SA	9AA	4NP	4NP
	Concentrations (μg/plate)	2.0	2.0	80	40	40
	Number of colonies/plate	790	583	922	2031	1631
	Medium	99	15	10	13	22
+	Vehicle	80	15	8	17	31
+	8	88	14	8	11	28
+	40	91	16	13	18	27
+	200	110	12	9	16	26
+	1000	83	12	14	17	27
+	5000	103	16	12	15	30
Positive controls + S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	5.0	2.5	2.5	5.0	5.0
	Number of colonies/plate	1339	84	215	1192	1069

SA: Sodium azide

9AA : 9-Aminoacridine

4NP : 4-Nitro-o-phenylenediamine

2-AA: 2-Aminoanthracene

 

The test substance was not genotoxic under the conditions tested.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative