2-Propenoic acid, 2-propylheptyl ester

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:

2008-02-14 to 2008-04-04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study according to OECD TG No.471 and in compliance with GLP regulations.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

21 Jul 1997

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-propylheptyl acrylate

EC Number:

604-669-5

Cas Number:

149021-58-9

Molecular formula:

C13 H24 O2

IUPAC Name:

2-propylheptyl acrylate

Test material form:

other: liquid

Details on test material:

- Name of test material: 2-Propylheptylacrylat rein
- Test substance number: 07/0846-1
- Analytical purity: 98.5 area-%
- Isomers composition: about 87 % 2 Propylheptylacrylate and 10 % 4-methyl-2-propylhexylacrylate
- Analytical report no.: 07L00384
- Lot/batch No.: B4112/13 - 03122007
- Expiration date of the lot/batch: 2008-12-03
- pH-value: Ca. 5.5 (undiluted test substance)

Method

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:

S9-Mix (Aroclor 1254-induced rat liver S9-fraction)

Test concentrations with justification for top dose:

331.3 μg - 5 300 μg/plate (Preincubation test)
21.2 μg - 5 300 μg/plate (Standard plate test)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of vehicle: Due to the limited solubility of the test substance in water, DMSO was used, which has been demonstrated to be suitable in bacterial reverse mutation tests and for which historical control data are available.

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (2-AA)

Remarks:

TA 1535, TA 100, TA 1537,TA 98, WP2 uvrA with S9 mix

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Remarks:

TA 1535, TA 100 without S9 mix

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylenediamine (NOPD)

Remarks:

TA 98 without S9 mix

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA 1537 without S9 mix

Controls 5

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

E.coli WP2 uvrA without S9 mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation
1st Experiment
Type of test: Standard plate test with and without S9 mix
2nd Experiment
Type of test: Preincubation test with and without S9 mix
3rd Experiment
Type of test: Preincubation test with and without S9 mix

- Preincubation period: 20 min
- Exposure duration: 48 - 72 h

NUMBER OF REPLICATIONS:
3 plates per dose or control


DETERMINATION OF CYTOTOXICITY
- Toxicity detected by a
• decrease in the number of revertants
• clearing or diminution of the background lawn (= reduced his- or trp- background growth)
• reduction in the titer
is recorded for all test groups both with and without S9 mix in all experiments.

Evaluation criteria:

Acceptance criteria
Generally, the experiment is considered valid if the following criteria are met:
- The number of revertant colonies in the negative controls was within the range of the historical negative control data for each tester strain.
- The sterility controls revealed no indication of bacterial contamination.
- The positive control substances both with and without S9 mix induced a distinct increase in the number of revertant colonies within the range of the historical positive control data or above.
- The titer of viable bacteria was >= 10E +08/mL.

The test chemical is considered positive in this assay if the following criteria are met:
- A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least one tester strain either without S9 mix or after adding a metabolizing system.

A test substance is generally considered non-mutagenic in this test if:
- The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in at least two experiments carried out independently of each other.

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Standard plate test: at 5 300 μg/plate. Preincubation test: from 2650 µg/plate onwards.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No
- Effects of osmolality: No
- Evaporation from medium: No
- Water solubility: Due to the limited solubility of the test substance in water, DMSO was used as vehicle.
- Precipitation: tested up to precipitation. Precipitation concentration was 5300 µg/plate in the standard plate test.

COMPARISON WITH HISTORICAL CONTROL DATA:
In this study with and without S9 mix, the number of revertant colonies in the negative controls was nearby or within the range of the historical negative control data for each tester strain.
The positive control substances both with and without S9 mix induced a significant increase in the number of revertant colonies within the range of the historical positive control data or above.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
A weak bacteriotoxic effect (slight decrease in the number of his+ revertants, slight reduction in the titer) was occasionally observed in the standard plate test depending on the strain and test conditions at 5300 μg/plate.
In the preincubation assay bacteriotoxicity (slight decrease in the number of his+ revertants, slight reduction in the titer) was occasionally observed depending on the strain and test conditions from about 2650 μg/plate onward.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Ames test, 1^stexperiment, Standard plate test:

Substance	Concentration (µg/plate)	TA1535		TA100		TA1537		TA98		E. coli
		-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
DMSO		1(*15)	1	1 (*102)	1	1 (*9)	1	1(*28)	1	1(*35)	1
Test substance	21.2	0.9	0.9	0.9	1.1	1.1	1	1	1	1.1	1
	106	0.7	0.9	1	0.9	0.8	1.1	0.9	1.2	1.1	1
	530	0.9	1.1	1	1.1	0.9	1	0.8	1.1	1.2	1
	2650	1.0	1.1	1.1	0.7	0.9	0.9	1	1.0	1	1
	5300 (P)	0.9	0.9	1.0	0.8	0.7	0.5	0.8	0.7	0.9	1
MNNG	5	37.2		9.1		44.5		19.7		16.9
2-AA	2.5		9.8		6.5		15.7		24.1		6.9

Ames test, 2^stexperiment, Preincubation test:

Substance	Concentration (µg/plate)	TA1535		TA100		TA1537		TA98		E. coli
		-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
DMSO		1(*20)	1	1(*99)	1	1(*7)	1(*11)	1(*15)	1	1(*41)	1
Test substance	331.2	0.7	0.9	1	0.9	2.2	0.7	1.4	1.6	0.7	1
	662.5	0.6	0.7	0.9	0.8	1.3	0.9	1.0	1.6	0.9	1
	1325	0.9	0.7	1.0	0.8	0.8	0.7	1.2	1.3	0.8	1
	2650	0.8	0.7	0.9	0.8	0.7	0.5	0.9	1	0.8	1
	5300	0.8	0.8	0.9	0.4	0.7	0.6	1.1	1	0.8	0.9
4-NQO	5	29.2		9.5		36.8		22.9		12.8
2-AA	60		4.2		6.4		8.1		38.1		4

Ames test, 3^stexperiment, Preincubation test:

Substance	Concentration (µg/plate)	TA1535		TA100		TA1537		TA98		E. coli
		-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
DMSO						1(*8)	1	1(*28)	1
Test substance	331.2					0.9	0.8	0.9	1
	662.5					1.1	0.8	1	0.8
	1325					1.1	0.8	1	1.1
	2650					1.1	1	0.8	1
	5300					0.6	0.6	0.9	0.8
NOPD	10					44		18.7
2-AA	2.5						14.6		19.2

*= number of revertants in Solvent control

P= precipitation of substance