1-Propene, hydroformylation products, high-boiling

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study (OECD guideline 471).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): EP202
- Physical state: Liquid, colorless, clear
- Analytical purity: The test substance is a mixture containing different components.
- Test substance No.: 06/0723-1
- Batch identification: F701501mH
- Stability under test conditions: The stability of the test substance under storage conditions throughout the study period is guaranteed.
- Storage condition of test material: Room temperature

Method

Target gene:

his- (S. typhimurium)/trp- (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

Metabolic activation:

with and without

Metabolic activation system:

Aroclor-induced rat liver S9 mix

Test concentrations with justification for top dose:

- standard plate test: 20.0 μg - 5000 μg/plate (all strains);
- preincubation test: 37.5 μg - 5000 μg/plate (TA 1535 with and without S9 mix); 18.8 μg - 5000 μg/plate (TA 100, without S9 mix); 37.5 μg - 5000 μg/plate (TA 100 with S9 mix); 6.3 μg - 2000 μg/plate (TA 1537 without S9 mix); 15.6 μg - 2000 μg/plate (TA 1537 with S9 mix); 62.5 μg - 5000 μg/plate (TA 98 without S9 mix); 31.3 μg - 5000 μg/plate (TA 98 with S9 mix); 312.5 μg - 5000 μg/plate (E. coli with and without S9 mix);

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Remarks:

(historical control data)

Positive controls:

yes

Positive control substance:

other: see Details on test system and conditions

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation;

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


DETERMINATION OF CYTOTOXICITY
- Method: decrease in the number of revertants, clearing or diminution of the background lawn (= reduced his- or trp- background growth), reduction in the titer


OTHER:
- positive controls: - with S9 mix: 2-aminoanthracene (2-AA), 2.5 μg/plate, dissolved in DMSO (TA 1535, TA 100, TA 1537, TA 98), 60 μg/plate, dissolved in DMSO (E. coli WP2 uvrA); - without S9 mix: N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 5 μg/plate, dissolved in DMSO (TA 1535, TA 100); 4-nitro-o-phenylendiamine (NOPD), 10 μg/plate, dissolved in DMSO (TA 98); 9-aminoacridine (AAC), 100 μg/plate, dissolved in DMSO (TA 1537); 4-nitroquinoline-N-oxide (4-NQO), 5 μg/plate, dissolved in DMSO (E. coli WP2 uvrA)

Evaluation 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 > 10^8/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

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:

other: A bacteriotoxic effect was observed in the standard plate test from about 2 500 μg/plate onward, in the preincubation assay from about 100 μg/plate onward (depending on the strain and test conditions).

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:

other: A bacteriotoxic effect was observed in the standard plate test from about 2 500 μg/plate onward, in the preincubation assay from about 100 μg/plate onward (depending on the strain and test conditions).

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

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

Standard plate test (20 - 5000 µg/plate)
Strain	Metabolic activation system	mean his^+/trp^+revertant colonies (negative control)	maximum revertant factor (conc. (µg/plate))	dose dependency	Assessment	maximum revertant factor (positive control)
TA 98	no	30	1.0 (20/100)	no	negative	27 (NQPD)
	yes	36	1.1 (100/500)	no	negative	16.5 (2-AA)
TA 100	no	104	1.0 (100/500)	no	negative	7.3 (MNNG)
	yes	110	1.0 (20/100)	no	negative	7.3 (2-AA)
TA 1537	no	11	0.9 (20/100)	no	negative	34.3 (AAC)
	yes	10	100 (500)	no	negative	13 (2-AA)
TA1535	no	15	1.2 (17)	no	negative	55.5 (MNNG)
	yes	16	1.0 (2500)	no	negative	7.6 (2-AA)
E. coli WP2 uvrA	no	44	1.2 (100)	no	negative	27.8 (4-NQO)
	yes	54	1.1 (100)	no	negative	3.2 (2-AA)
reduced background growth/precipitation at 5000 µg/plate in all strains tested
Preincubation test, experiment 1
Strain	Metabolic activation system	mean his^+/trp^+revertant colonies (control)	maximum revertant factor (conc. (µg/plate))	dose dependency	Assessment	maximum revertant factor (positive control)	dose interval (µg/plate)
TA 98*	no	28	1.0 (312.5)	no	negative	30.3 (NQPD)	312.5 - 5000
	yes	46	0.7 (312.5)	no	negative	12.1 (2-AA)
TA 100*	no	124	0.5 (312.5)	no	negative	7.7 (MNNG)	312.5 - 5000
	yes	136	0.7 (312.5)	no	negative	5.5 (2-AA)
TA 1537**	no	13	0.1 (125)	no	negative	37.7 (AAC)	125 - 2000
	yes	10	1.1 (125)	no	negative	11.3 (2-AA)
TA 1535*	no	19	0.8 (312.5)	no	negative	62.3 (MNNG)	312.5 - 5000
	yes	19	0.7 (312.5)	no	negative	7 (2-AA)
E. coli WP2 uvrA***	no	41	1.0 (312.5)	no	negative	13.4 (4-NQO)	312.5 - 5000
	yes	46	1.0 (625)	no	negative	4.7 (2-AA)
* > 312.5 µg/plate: reduced background growth; 5000 µg/plate:reduced background growth/precipitation
**>/= 125 µg/plate: reduced background growth
*** 5000 µg/plate:reduced background growth/precipitation
Preincubation test, experiment 2
Strain	Metabolic activation system	mean his^+revertant colonies (control)	maximum revertant factor (conc. (µg/plate))	dose dependency	Assessment	maximum revertant factor (positive control)	dose interval (µg/plate)
TA 98****	no	32	1.1 (125)	no	negative	17.3 (NQPD)	62.5 - 1000
	yes	31	1.2 (31.3/62.5)	no	negative	17.3 (2-AA)	31.3 - 500
TA 100**	no	100	1.1 (75)	no	negative	9.5 (MNNG)	18.8 - 300
	yes	107	1.0 (37.5/75/150)	no	negative	7.3 (2-AA)	37.5 - 600
TA 1537***	no	10	1.0 (12.5)	no	negative	34.7 (AAC)	6.3 - 100
	yes	9	1.3 (31.3)	no	negative	15.5 (2-AA)	15.6 - 250
TA 1535*	no	17	1.0 (16)	no	negative	55.4 (MNNG)	37.5 - 600
	yes	17	0.9 (37.5/150)	no	negative	6.8 (2-AA)
* 600 µg/plate: reduced background growth
** 300 µg/plate (without S9) / 600 µg/plate (with S9): reduced background growth
*** 100 µg/plate (without S9) / 250 µg/plate (with S9): reduced background growth
**** 1000 µg/plate (without S9) / 500 µg/plate (with S9): reduced background growth

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

According to the results of the present study, the test substance EP202 is not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay under the experimental conditions chosen here.

Executive summary:

The mutagenic potential of EP202 was determined in the Salmonella typhimurium/Escherichia coli reverse mutation assay. Under the chosen conditions, the test substance EP202 was not mutagenic.