Fatty acids, C16 and C16-18-unsatd., mixed esters with adipic acid and pentaerythritol

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:

26 July - 09 Aug 2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP - Guideline study. According to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", 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:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(adopted July 1997)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

(2000/32/EC)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Comunidad de Madrid

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane

EC Number:

306-085-3

EC Name:

Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane

Cas Number:

95912-89-3

IUPAC Name:

95912-89-3

Details on test material:

- Name of test material (as cited in study report):Trade name given
- Analytical purity: 100%
- Physical state: liquid, orange/brown
- Batch No.: ES090146
- Storage condition of test material: room temperature

Method

Target gene:

his operon (S. typhimurium) and trp operon (E. coli)

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 pKM 101

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:

0.06, 0.19, 0.56, 1.67 and 5 µL/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: corn oil

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-amino-anthracene (2AA; 1.5 µg/plate in DMSO, +S9, TA98, 2.5 µg/plate in DMSO, +S9, TA100 and TA1537, 30 µg/plate in DMSO, +S9, TA1535 and WP2)

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

Migrated to IUCLID6: (2NF; 5 µg/plate in DMSO, -S9, TA98)

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Migrated to IUCLID6: (SA; 2.5 µg/plate in H2O, -S9, TA100; 3.5 µg/plate in H2O, -S9, TA1537)

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

Migrated to IUCLID6: (4NQO; 0.4 µg/plate in DMSO, -S9, WP2)

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

Migrated to IUCLID6: (9AA, 45 µg/plate in DMSO, -S9, TA1537)

Details on test system and experimental conditions:

METHOD OF APPLICATION: plate incorporation (main study) ; preincubation (confirmatory study)
DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: triplicates each in two independent experiments

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

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 (TA98, TA100, WP2) or three-fold (TA1535, TA1537) 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 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:

not specified

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A pKM 101

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

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Test Results of Experiment 1 (plate incorporation)

With or without S9-Mix	Test substance concentration (μL/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	WP2	TA98	TA1537
–	0	103	23	184	22	6
–	0.06	110	25	188	17	4
–	0.19	94	30	222	20	5
–	0.56	91	24	212	18	5
–	1.67	95	19	207	18	4
–	5	93	17	232	18	5
Positive controls, –S9	Name	SA	SA	4NQO	2NF	9AA
	Concentrations (μg/plate)	2.5	3.5	0.4	5	45
	Mean No. of colonies/plate (average of 3)	762	798	1776	418	188
+	0	92	23	234	22	4
+	0.06	87	21	208	20	5
+	0.19	90	17	203	17	6
+	0.56	95	18	207	20	6
+	1.67	92	21	212	20	7
+	5	97	23	213	22	8
Positive controls, + S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	2.5	30	30	1.5	2.5
	Mean No. of colonies/plate (average of 3)	1677	419	1605	1012	190

4NQO = 4-nitroquinoline-N-oxide

9AA = 9-aminoacridine

2AA = 2-aminoanthracene

2NF = 2-nitroflurene

SA = sodium azide

 

 

Table 2: Test Results of Experiment 2 (pre-incubation)

With or without S9-Mix	Test substance concentration (μL/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	WP2	TA98	TA1537
–	0	83	24	138	23	5
–	0.06	87	19	187	18	7
–	0.19	79	27	194	17	7
–	0.56	85	24	170	19	4
–	1.67	89	22	181	23	5
–	5	91	19	193	20	5
Positive controls, –S9	Name	SA	SA	4NQO	2NF	9AA
	Concentrations (μg/plate)	2.5	3.5	0.4	5	45
	Mean No. of colonies/plate (average of 3)	792	1036	2482	409	161
+	0	82	33	163	26	9
+	0.06	91	23	199	21	6
+	0.19	96	26	219	20	7
+	0.56	95	27	182	19	9
+	1.67	83	26	208	22	6
+	5	91	23	208	20	8
Positive controls, + S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	2.5	30	30	1.5	2.5
	Mean No. of colonies/plate (average of 3)	1007	320	2118	1049	189

4NQO = 4-nitroquinoline-N-oxide

9AA = 9-aminoacridine

2AA = 2-aminoanthracene

2NF = 2-nitroflurene

SA = sodium azide

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative