Isooctadecanoic acid, monoester with glycerol

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:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP-Guideline study. Tested with the source substance 2,3-dihydroxypropyl oleate. According to the ECHA guidance document 'Practical guide 6: How to report read-across and categories (ECHA, 2012)', the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,3-dihydroxypropyl oleate

EC Number:

203-827-7

EC Name:

2,3-dihydroxypropyl oleate

Cas Number:

111-03-5

IUPAC Name:

2,3-dihydroxypropyl octadec-9-enoate

Details on test material:

- Name of test material (as cited in study report): glycerol monooleate
- Physical state: light yellow pellets with characteristic odour
- Analytical purity: 99.93%
- Stability under test conditions: verified after test

Method

Target gene:

his operon (for S. typhimurium strains)
trp operon (for E. coli strain)

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:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with phenobarbital and 5,6-benzoflavone

Test concentrations with justification for top dose:

-S9 mix:
4.88, 9.77, 19.5, 39.1, 78.1, 156 μg/plate (TA100, TA98)
156, 313, 625, 1250, 2500, 5000 μg/plate (TA1535, WP2 uvrA)
0.153, 0.305, 0.610, 1.22, 2.44, 4.88, 9.77 μg/ plate (TA1537)

+S9 mix:
9.77, 19.5, 39.1, 78.1, 156, 313, 625 μg/plate (TA100)
156, 313, 625, 1250, 2500, 5000 μg/plate (TA1535, WP2 uvrA, TA98)
9.77, 19.5, 39.1, 78.1, 156, 313 μg/plate (TA1537)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test substance was not soluble in water.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: -S9: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (AF2; 0.01 or 0.1 µg/plate, TA100, TA98, E. coli); sodium azide (SA; 0.5 µg/plate, TA1535); 9-aminoacridine (9AA; 80 µg/plate, TA1537); +S9: 2-Aminoanthracene (2AA; 0.5-10 µg/plate, all strains)

Details on test system and experimental conditions:

METHOD OF APPLICATION: Pre-incubation method

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 replications in one experiment

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

Evaluation criteria:

Revertant colonies were counted for 3 plates per bacterial strain. The test result was considered positive, if the mean value out of 3 plates was at least twice as high as the negative vehicle control and a dose-dependency was found.

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:

cytotoxicity

Remarks:

-S9: at 3.13 (TA1537), 78.1 (TA98) and 100 (TA100) µg/plate and above; +S9: at 128 (TA1537) and 156 (TA100) µg/plate and above

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: the test substance was not soluble in water.
- Precipitation: in the dose-finding studies and the main study, precipitation was observed at the end of the exposure period at ≥ 625 and ≥ 1250 µg/plate in the presence and absence of metabolic activation, respectively.

RANGE-FINDING/SCREENING STUDIES:
The highest concentrations analysed in the main study were selected based on the solubility and/or cytotoxicity of the test substance in the cell culture medium. Three dose-finding studies were performed:

[Dose-finding study]
-S9 mix: 8.19, 20.5, 51.2, 128, 320, 800, 2000, 5000 μg/plate (all strains)
+S9 mix: 8.19, 20.5, 51.2, 128, 320, 800, 2000, 5000 μg/plate (all strains)
[Dose-finding restudy]
-S9 mix: 8.19, 20.5, 51.2, 128, 320, 800, 2000, 5000 μg/plate (all strains)
+S9 mix: 8.19, 20.5, 51.2, 128, 320, 800, 2000, 5000 μg/plate (all strains)
[Dose-finding additional study]
-S9 mix
6.25, 12.5, 25.0, 50.0, 100, 200 μg/plate (TA100),
3.13, 6.25, 12.5, 25.0, 50.0, 100 μg/plate (TA98),
0.0977, 0.195, 0.391, 0.781, 1.56, 3.13, 6.25, 12.5 μg/plate (TA1537)
+S9 mix:
6.25, 12.5, 25.0, 50.0, 100, 200 μg/plate (TA1537)

ADDITIONAL INFORMATION ON CYTOTOXICITY:
The test substance was not cytotoxic to tester strains TA1535 and WP2uvrA at any concentration with and without metabolic activation.
In the dose-finding studies and in the main study, the minimum cytotoxic concentrations determined for the tester strains TA100, TA98 and TA1537 were as follows:

[Dose-finding study]
-S9 mix: 8.19 (TA1537) and 128 (TA100) μg/plate
+S9 mix: 128 (TA1537) and 320 (TA100) μg/plate
[Dose-finding restudy]
-S9 mix: 8.19 (TA1537), 51.2 (TA98) and 128 (TA100) μg/plate
+S9 mix: 128 (TA1537), 320 (TA100) μg/plate
[Dose-finding additional study]
-S9 mix: 3.13 (TA1537) and 100 (TA100, TA98) μg/plate
+S9 mix: 200 (TA1537) μg/plate
[Main study]
-S9 mix: 4.88 (TA1537), 78.1 (TA98) and 156 (TA100) µg/plate
+S9 mix: 156 (TA100) and 313 (TA1537) µg/plate

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1. Results of the bacterial reverse mutation test

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
		TA 100	TA1535	WP2urvA	TA98	TA1537
–	0	129 ± 7	13 ± 2	25 ± 5	21 ± 2	9 ± 1
–	0.153					9 ± 2
–	0.305					7 ± 1
–	0.610					9 ± 1
–	1.22					12 ± 1
–	2.44					7 ± 2
–	4.88	129 ± 5			21 ± 1	10 ± 2*
–	9.77	139 ± 4			18 ± 3	10 ± 3*
–	19.5	125 ± 5			20 ± 4
–	39.1	134 ± 3			17 ± 2
–	78.1	142 ± 6			18 ± 3*
–	156	131 ± 6 *	12 ± 2	16 ± 3	24 ± 6*
–	313		10 ± 3	22 ± 3
–	625		12 ± 2	21 ± 4
–	1250 (+)		10 ± 2	16 ± 2
–	2500 (+)		10 ± 2	16 ± 2
–	5000 (+)		9 ± 1	13 ± 3
Positive controls, –S9	Name	AF2	SA	AF2	AF2	9AA
	Concentrations (μg/plate)	0.01	0.5	0.01	0.1	80
	Mean No. of colonies/plate (average of 3 plates ± SD)	886 ± 5	549 ± 8	148 ± 19	689 ± 53	200 ±7
+	0	121 ± 4	13 ± 1	26 ± 5	29 ± 1	25 ± 3
+	9.77	132 ± 7				20 ± 2
+	19.5	130 ± 6				20 ± 2
+	39.1	125 ± 8				21 ± 2
+	78.1	121 ± 4				13 ± 4
+	156	123 ± 4*	14 ± 0	25 ± 4	24 ± 3	12 ± 2
+	313	118 ± 5*	12 ± 2	19 ± 5	25 ± 3	10 ± 2*
+	625 (+)	106 ± 8*	13 ± 2	23 ± 4	26 ± 6
+	1250 (+)		8 ± 2	16 ± 2	19 ± 3
+	2500 (+)		9 ± 1	16 ± 2	19 ± 1
+	5000 (+)		11 ± 1	16 ± 2	12 ± 2
Positive controls, +S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	1	2	10	0.5	2
	Mean No. of colonies/plate (average of 3 plates ± SD)	1018 ± 35	336 ± 19	732 ± 65	370 ± 10	142 ± 5

(+): Visible precipitation was observed at the end of exposure period.

*: Growth inhibition was observed

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative