Fatty acids, C7-9, tetraesters with pentaerythritol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

8. Dec. 1998 - 19. Jan. 1999

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see "remarks"

Remarks:

GLP-Guideline study. According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (Dec 2012)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Justification for type of information:

refer to analogue justification provided in IUCLID section 13

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

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:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Safepharm Laboratories Limited, Derby, UK

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

131459-39-7

Cas Number:

131459-39-7

IUPAC Name:

131459-39-7

Method

Target gene:

his operon and trp operon

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: incapable of DNA excision repair (uvrB-); increased permeability of the cell wall (rfa)

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 Sprague-Dawley rats treated with Aroclor 1254

Test concentrations with justification for top dose:

50, 150, 500, 1500, 5000 µg/plate (first and second experiment)

Vehicle / solvent:

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

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: without S9: N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG; 3 µg/plate TA100, 5 µg/plate TA1535, 2 µg/plate WP2uvrA-), 9-Aminoacridine (9-AA; 80 µg/plate TA 1535), 4-Nitroquinoline-1 (4NQO; 0.2 µg/plate TA98)

Controls 2

Positive controls:

yes

Positive control substance:

other: with S9 mix: 2-Aminoanthracene (2AA; 1 µg/plate TA 100, 2 µg/plate TA 1535 and TA 1537, 10 µg/plate WP2uvrA-); Benzo(a)pyrene (BP; 5 µg/plate TA 98)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 replications each in 2 independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: inspection of bacterial background lawn, number of revertant colonies

Evaluation criteria:

The test material may be considered to be positive in this test system if the following criteria are met: All tester strain cultures exhibit a characteristic number of spontaneous revertants per plate in the vehicle and untreated controls (see table 1); Each positive control value was at least two times the respective vehicle control value for each strain (for historical values see table 2); A minimum of 4 non-toxic dose levels were achieved; The test material should have induced a reproducible, dose-related and statistically significant increase in the revertant count in at least one strain of bacteria.

Statistics:

Dunett's method of linear regression, Kirkland DJ (1989), Statistical Evaluation of Mutagenicity Test Data. UKEMS Sub-comittee on Guidelines for Mutagenicity Testing. Report - Part III - Cambridge University Press

Results and discussion

Test results

Test results 1

Species / strain:

other: 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, but tested up to precipitating concentrations

Remarks:

oily precipitate at 5000 µg/plate which did not prevent scoring of revertant colonies

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, but tested up to precipitating concentrations

Remarks:

oily precipitate at 5000 µg/plate which did not prevent scoring of revertant colonies

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: oily precipitate at 5000 µg/plate

RANGE-FINDING/SCREENING STUDIES: the dose range of the test material used in the preliminary toxicity study was 0, 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500, 5000 µg/plate. The test material was non-toxic to the strains of bacteria used (TA100 and WP2uvrA-).

COMPARISON WITH HISTORICAL CONTROL DATA: Values of positive and negative controls were in range of historical control data.

Any other information on results incl. tables

Table 3. Test results of experiment 1 (plate incorporation).

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	WP2uvrA-	TA98	TA1537
–	0	74 ± 9.6	21 ± 2.3	22 ± 2.0	23 ± 4.0	8 ± 2.0
–	50	75 ± 7.1	20 ± 2.1	24 ± 2.1	24 ± 4.0	6 ± 2.0
–	150	79 ± 13.1	20 ± 6.8	22 ± 1.5	19 ± 2.6	10 ± 1.0
–	500	76 ± 4.2	15 ± 4.0	23 ± 7.0	21 ± 4.9	9 ± 3.8
–	1500	71 ± 6.1	20 ± 3.8	18 ± 5.5	21 ± 5.7	10 ± 3.5
–	5000	73 ± 1.5 P	21 ± 5.5 P	24 ± 6.0 P	21 ± 5.9 P	9 ± 2.5 P
Positive controls, –S9	Name	ENNG	ENNG	ENNG	4NQO	9AA
	Concentrations (μg/plate)	3	5	2	0.2	80
	Mean No. of colonies/plate (average of 3 ± SD)	323 ± 11.7	283 ± 10.0	503 ± 24.2	166 ± 5.3	1027 ± 358.2
+	0	87 ± 1.5	16 ± 1.5	28 ± 2.3	34 ± 6.7	18 ± 0.6
+	50	94 ± 6.1	13 ± 1.2	29 ± 5.8	31 ± 0.6	23 ± 7.4
+	150	88 ± 4.0	13 ± 2.6	25 ± 5.6	35 ± 6.4	19 ± 7.0
+	500	87 ± 3.5	12 ± 1.2	29 ± 3.8	32 ± 9.5	20 ± 5.5
+	1500	81 ± 6.5	10 ± 0.6	30 ± 3.6	33 ± 3.8	15 ± 4.6
+	5000	77 ± 5.1 P	12 ± 0.6 P	30 ± 2.6 P	31 ± 4.7 P	18 ± 3.8 P
Positive controls, +S9	Name	2AA	2AA	2AA	BP	2AA
	Concentrations (μg/plate)	1	2	10	5	2
	Mean No. of colonies/plate (average of 3 ± SD)	920 ± 153.0	198 ± 9.3	555 ± 27.8	511 ± 132.0	196 ± 6.5

ENNG = N-ethyl-N-nitro-N-nitrosoguanidine

4NQO = 4-nitroquinoline-N-oxide

9AA = 9-aminoacridine

2AA = 2 -aminoanthracene

BP = Benzo(a)pyrene

P = Precipitate

Table 4. Test results of experiment 2 (plate incorporation).

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	WP2uvrA-	TA98	TA1537
–	0	103 ± 5.3	26 ± 3.8	31 ± 8.5	29 ± 7.0	12 ± 2.9
–	50	105 ± 10.8	21 ± 4.9	30 ± 7.4	28 ± 7.1	11 ± 1.0
–	150	97 ± 7.0	26 ± 1.0	35 ± 2.9	22 ± 1.2	13 ± 3.5
–	500	104 ± 9.1	25 ± 4.6	34 ± 5.1	26 ± 5.6	15 ± 1.7
–	1500	110 ± 9.5	23 ± 6.0	25 ± 0.6	27 ± 3.8	13 ± 1.7
–	5000	112 ± 5.7 P	28 ± 0.6 P	35 ± 2.6 P	20 ± 1.0 P	11 ± 1.5 P
Positive controls, –S9	Name	ENNG	ENNG	ENNG	4NQO	9AA
	Concentrations (μg/plate)	3	5	2	0.2	80
	Mean No. of colonies/plate (average of 3 ± SD)	880 ± 31.5	257 ± 34.4	1191 ± 59.3	166 ± 4.6	1384 ± 263.4
+	0	103 ± 8.6	13 ± 3.6	37 ± 2.1	27 ± 6.2	20 ± 3.8
+	50	92 ± 21.0	12 ± 1.5	33 ± 4.0	27 ± 7.2	20 ± 2.6
+	150	91 ± 7.5	14 ± 1.5	34 ± 2.6	29 ± 4.0	22 ± 2.6
+	500	113 ± 15.0	14 ± 2.3	39 ± 2.1	27 ± 6.7	15 ± 3.2
+	1500	110 ± 7.6	13 ± 2.6	36 ± 3.2	31 ± 4.9	21 ± 3.1
+	5000	97 ± 9.0 P	17 ± 5.8 P	30 ± 2.5 P	28 ± 4.7 P	22 ± 3.0 P
Positive controls, +S9	Name	2AA	2AA	2AA	BP	2AA
	Concentrations (μg/plate)	1	2	10	5	2
	Mean No. of colonies/plate (average of 3 ± SD)	865 ± 43.2	246 ± 7.5	1051 ± 90.0	611 ± 51.5	182 ± 7.0

ENNG = N-ethyl-N-nitro-N-nitrosoguanidine

4NQO = 4-nitroquinoline-N-oxide

9AA = 9-aminoacridine

2AA = 2 -aminoanthracene

BP = Benzo(a)pyrene

P = Precipitate

Applicant's summary and conclusion

Conclusions:

negative