Methyl myristate

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:

key study

Study period:

1992-03-25 to 1992-04-02

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

, analytical purity of test material not specified; incomplete strain selection;

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Methyl laurate

EC Number:

203-911-3

EC Name:

Methyl laurate

Cas Number:

111-82-0

Molecular formula:

C13H26O2

IUPAC Name:

methyl laurate

Details on test material:

- Name of test material (as cited in study report): Lauric acid methyl ester
- Substance type: Fatty acid methyl ester
- Physical state: colourless liquid
- Analytical purity: not given
- Storage condition of test material: RT

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 TA 100)

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 (Wistery 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: acetone
To prevent toxic effects of the solvent medium the stock solution of the test substance was doubled and 50 µL/plate instead of 100 µl/plate was introduced in the test

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

culture media

Negative solvent / vehicle controls:

yes

Remarks:

Acetone

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA 100 and TA 1535 without metabolic activation

Controls 2

Untreated negative controls:

yes

Remarks:

culture media

Negative solvent / vehicle controls:

yes

Remarks:

Acetone

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA 1537 without metabolic activation

Controls 3

Untreated negative controls:

yes

Remarks:

culture media

Negative solvent / vehicle controls:

yes

Remarks:

Acetone

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylendiamine

Remarks:

TA 98 and TA 1538 without metabolic activation

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

For all strainn with metabolic activation (S9 mix, Aroclor induced)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

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

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

NUMBER OF REPLICATIONS: triplicates

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

Evaluation criteria:

The test material may be considered positive in this test system if the following criteria are met:
For the test item to be considered mutagenic, two-fold (or more) increases in mean revertant numbers must be observed at two consecutive dose levels or at the highest practicable dose level only. In addition, there must be evidence of a dose-response relationship showing increasing numbers of mutant colonies with increasing dose levels.
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:

cytotoxicity

Remarks:

at 5000 µg/plate

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:

cytotoxicity

Remarks:

at 5000 µg/plate

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 (¿g/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		TA 1535	TA100	TA1537	TA1538	TA98
	Buffer	14	132	12	9	21
-	Solvent (Acetone)	15	133	10	13	26
-	8	11	122	9	6	18
-	40	10	96	8	12	16
-	200	10	109	8	11	16
-	1000	9	92	13	11	18
	5000	10	122	10	11	13
Positive controls - S9	Name	SA	SA	9AA	4ND	4ND
	Concentrations (¿g/plate)	2	2	80	40	40
	Number of colonies/plate	590	659	771	2408	1813
+	Buffer	16	158	11	28	35
+	Solvent (Acetone)	17	153	7	32	41
+	8	16	126	13	34	24
+	40	13	209	8	32	22
	200	13	102	9	31	28
+	1000	13	112	7	18	24
+	5000	7	70	5	16	18
Positive controls + S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (¿g/plate)	2.5	5	2.5	5	5
	Number of colonies/plate	112	1532	137	1146	1555

 

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 (¿g/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		TA 1535	TA100	TA1537	TA1538	TA98
	Buffer	20	133	10	6	26
-	Solvent (Acetone)	20	123	12	6	16
-	8	16	128	9	8	21
-	40	16	115	7	6	15
-	200	13	109	7	8	21
-	1000	14	132	13	9	19
	5000	17	110	11	11	28
Positive controls - S9	Name	SA	SA	9AA	4ND	4ND
	Concentrations (¿g/plate)	2	2	80	40	40
	Number of colonies/plate	465	652	978	2055	1816
+	Buffer	18	125	11	23	42
+	Solvent (Acetone)	23	102	7	23	35
+	8	27	137	7	13	43
+	40	17	125	5	24	31
	200	21	119	5	22	30
+	1000	19	105	8	18	27
+	5000	6	39	4	12	22
Positive controls + S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (¿g/plate)	2.5	5	2.5	5	5
	Number of colonies/plate	104	1127	137	738	831

 

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.