Reaction mass of 1,2-Diacetoxy-3-butene and 1,2-Dihydroxy-3-butene, monoacetate

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:

2002-11-21 - 2003-05-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline Study (OECD)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

445-630-0

EC Name:

-

Molecular formula:

Unspecified

IUPAC Name:

1-(acetyloxy)but-3-en-2-yl acetate; 2-hydroxybut-3-en-1-yl acetate

Details on test material:

- Name of test material (as cited in study report): Vinylglykoldiacetat 75% ig
- Test substance No.: 02/0526-1
- Date of manufacture: October 21, 2001
- Physical state: Colorless liquid
- Analytical purity: 74.8 area %
- Lot/batch No.: Kanister 1, 2, 4, 5
- Storage condition of test material: Room temperature (N2 conditions)

Method

Target gene:

Histidine operon

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 1254-induced rat liver S9-mix

Test concentrations with justification for top dose:

Standard plate test: 0, 27, 135, 675, 3375, 6750 µg/plate
Preincubation test: 0, 5.4, 27, 135, 675, 3.375 µg/plate

Vehicle / solvent:

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

Controls

Controls 1

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene; 2.5 µg/plate, dissolved in DMSO for TA 1535, TA 100, TA 1537, TA 98; 60 µg/plate, dissolved in DMSO for E. coli WP2 uvrA

Remarks:

with metabolic activation

Controls 2

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: N-methyl-N'-nitro-N-nitrosoguanidine (MNNG); 5 µg/plate, dissolved in DMSO for strains TA 1535, TA 100

Remarks:

without metabolic activation

Controls 3

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylendiamine (NOPD); 10 µg/plate, dissolved in DMSO for strain TA 98

Remarks:

without metabolic activation

Controls 4

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

without metabolic activation

Migrated to IUCLID6: 100 µg/plate, dissolved in DMSO for strain TA 1537

Controls 5

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

without metabolic activation

Migrated to IUCLID6: 5 µg/plate, dissolved in DMSO for strain E. coli WP2 uvrA

Details on test system and experimental conditions:

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

DURATION
- Preincubation period: 20 min, 37°C
- Exposure duration: 48 - 72 h, 37°C

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: decrease in the number of revertants, clearing or diminution of the background lawn, reduction in the titer

Evaluation criteria:

- 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 S-9 mix or after adding a metabolizing system.
- A test substance is generally considered nonmutagenic in this test if:
The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in 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:

cytotoxicity

Vehicle controls validity:

valid

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:

cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No test substance precipitation was found

RANGE-FINDING/SCREENING STUDIES:
The experimental procedure was based on the method described by Yahagi et al. (1977) and
Matsushima et al . (1980).
0.1 ml test solution or vehicle, 0.1 ml bacterial suspension and 0 .5 ml S-9 mix or phosphate buffer were incubated at 37'C for the duration of about 20 minutes using a shaker. Subsequently, 2 ml of soft agar was added and, after mixing, the samples are poured onto the agar plates within approx. 30 seconds. After incubation at 37'C for 48 - 72 hours in the dark, the bacterial colonies were counted.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
A weak bacteriotoxic effect (slight decrease in the number of revertants, slight reduction
in the titer) was observed in the standard plate test depending on the strain and test
conditions at doses > 3,375 pg/plate .
In the preincubation assay bacteriotoxicity (reduced background growth, slight decrease in the number of revertants, slight reduction in the titer) was observed depending on the strain and test conditions at doses > 675 pg/plate.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Results standard plate experiment:

Dose µg/plate	metabolic activation	TA98			TA100			TA1535			TA1537			E.coli WP2 uvrA
		Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3
0	+	44	40	34	102	111	106	16	14	19	12	8	12	28	32	34
27	+	41	37	37	110	105	100	17	14	21	16	14	11	30	32	30
135	+	39	39	36	106	104	110	23	19	20	15	11	11	20	31	26
675	+	30	33	36	102	91	103	18	15	17	7	7	7	29	24	31
3375	+	26	37	14	95	90	94	14	18	14	11	12	10	20	c	24
6750	+	17	16	16	70	53	49	8	11	8	6	7	4	17	16	19
2.5 µg 2-Aminoanthracene	+	864	899	838	902	931	874	190	134	146	111	124	131
60 µg 2-Aminoanthracene	+													250	264	134
0	-	28	25	26	100	107	103	18	14	17	9	9	10	25	35	30
27	-	20	28	27	92	107	101	16	17	13	8	3	7	34	26	26
135	-	26	27	26	109	104	106	16	19	19	10	4	7	28	34	31
675	-	26	31	21	104	107	95	20	12	14	5	8	6	24	25	26
3375	-	17	12	15	102	100	91	17	16	12	10	10	7	26	20	26
6750	-	1	1	1	26	17	25	4	4	8	2	4	1	17	19	19
5 µg MNNG	-				742	713	792	612	621	658
10 µg 4-Nitro-o-phenylendiamin	-	613	609	572
100 µg 9-Aminoacridiniumchloride monohydrate	-										431	381	417
5 µg 4-nitroquinoline-N-oxide	-													250	264	134

Results preincubation experiment:

Dose µg/plate	metabolic activation	TA98			TA100			TA1535			TA1537			E.coli WP2 uvrA
		Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3
0	+	38	40	34	110	102	112	15	16	19	12	10	7	29	35	35
5.4	+	39	43	36	101	90	114	14	11	13	11	8	8	28	21	33
27	+	23	24	36	109	103	77	20	14	12	11	18	12	36	24	21
135	+	33	31	45	85	109	84	14	18	13	10	9	6	28	22	17
675	+	33	31	29	68	97	91	9	7	8	11	6	11	26	21	14
3375	+	10b	14b	8b	86b	72b	68b	11b	6b	8b	5b	5b	7b	12b	26b	28b
2.5 µg 2-Aminoanthracene	+	882	890	813	688	763	734	102	97	90	131	140	128
60 µg 2-Aminoanthracene	+													255	211	270
0	-	22	34	34	102	103	107	17	16	12	9	13	10	36	32	30
5.4	-	30	31	27	94	105	115	13	19	13	10	7	9	37	31	20
27	-	30	22	26	92	102	108	15	19	19	7	11	10	27	34	29
135	-	22	19	27	105	101	108	17	17	12	7	6	10	22	32	30
675	-	20	20	23	104	110	120	13	13	19	9	8	9	30	31	29
3375	-	0b	0b	0b	13b	25b	30b	0b	0b	0b	0b	0b	0b	20b	12b	14b
5 µg MNNG	-				1024	887	1149	820	917	754
10 µg 4-Nitro-o-phenylendiamin	-	954	930	892							497	382	426
100 µg 9-Aminoacridiniumchloride monohydrate	-
5 µg 4-nitroquinoline-N-oxide	-													544	632	512

c: contamination

b: reduced backround growth