Fatty acids, rape-oil, Me esters

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:

February 10th, 1988 - February 26th, 1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1988

Report date:

1988

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

, incomplete strain selection; lack of details on test substance specification;

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C14-18 and C16-18-unsatd., Me esters

EC Number:

267-007-0

EC Name:

Fatty acids, C14-18 and C16-18-unsatd., Me esters

Cas Number:

67762-26-9

Molecular formula:

UVCB substance, not univocal molecular formula available

IUPAC Name:

Fatty acids, C14-18 and C16-18 unsaturated, methyl esters

Details on test material:

- Name of test material: Methyl Oleate
- Physical state: milky white, waxy

Method

Target gene:

Genes involved in Histidine synthesis

Species / strain

Species / strain 1

Species / strain / cell type:

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

Additional strain / cell type characteristics:

other: rfa-; uvrB- (R+ for TA 98 and TA100)

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Additional strain / cell type characteristics:

other: rfa-; uvrB-

Metabolic activation:

with and without

Metabolic activation system:

S9-Mix (Spraque Dawley rats, male, Aroclor 1254 induced)

Test concentrations with justification for top dose:

0, 8, 40, 200, 1000 and 5000 µg/plate in both experiments

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Tween 80/bidest. water

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA1535 and TA100 without metabolic activation

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA1537 without metabolic activation

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: 4-Nitro-o-phenyldiamin

Remarks:

TA1538 and TA98 without metabolic activation

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracen

Remarks:

All test strains with metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation)

DURATION
- Exposure duration: 48 h
- Expression time (cells in growth medium): 48 h

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: Inspection of the bacterial background lawn


OTHER: The spontaneous mutation rates of each tester strain were within the characteristic spontaneous mutation rates

Evaluation criteria:

Revertant colonies were counted and the mean and standard deviation were calculated and compared to the controls. The tester strains were regulary tested for their genetic markers.
All cultures must demonstrate the characteristic mean number of spontaneous revertants in the vehicle controls.

Statistics:

Mean 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

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

Positive controls validity:

valid

Additional information on results:

COMPARISON WITH HISTORICAL CONTROL DATA: Yes

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Mutagenicity on bacteria - experiment I

 

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		TA 1535	TA100	TA1537	TA1538	TA98
-	Solvent (Tween80/H2O)	5	109	6	12	13
-	8	6	108	8	3	14
-	40	4	107	4	7	12
-	200	7	101	5	5	15
-	1000	5	111	9	8	14
	5000	7	120	9	12	16
Positive controls - S9	Name	SA	SA	9AA	4ND	4ND
	Concentrations (μg/plate)	2	2	80	40	40
	Number of colonies/plate	241	420	156	892	351
+	Solvent (Tween80/H2O)	11	114	7	13	17
+	8	11	112	5	11	16
+	40	12	112	6	10	15
	200	6	115	4	8	18
+	1000	10	127	4	7	18
+	5000	10	119	5	8	12
Positive controls + S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	2.5	5	2.5	5	5
	Number of colonies/plate	117	1316	170	187	826

 

9AA = 9-Aminoacridine

2AA = 2-Aminoanthracene

SA = Sodium Acide

4ND = 4-Nitro-o-phenylendiamine

 

 

Table 2: Mutagenicity on bacteria - experiment II

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		TA 1535	TA100	TA1537	TA1538	TA98
-	Solvent (Tween80/H2O)	10	81	5	7	18
-	8	7	87	6	9	22
-	40	7	90	5	6	17
-	200	12	97	5	11	21
-	1000	8	93	6	10	19
	5000	7	87	8	7	20
Positive controls - S9	Name	SA	SA	9AA	4ND	4ND
	Concentrations (μg/plate)	2	2	80	40	40
	Number of colonies/plate	251	262	230	1038	401
+	Solvent (Tween80/H2O)	11	91	7	12	16
+	8	14	96	10	11	20
+	40	9	109	7	13	23
	200	18	105	7	13	17
+	1000	13	107	8	13	20
+	5000	11	101	6	18	12
Positive controls + S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	2.5	5	2.5	5	5
	Number of colonies/plate	131	1285	51	366	618

 

9AA = 9-Aminoacridine

2AA = 2-Aminoanthracene

SA = Sodium Acide

4ND = 4-Nitro-o-phenylendiamine

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative Under the tested experimental conditions the test substance did not induce gene mutations in S. typhimurium strains up to the maximum dose.