3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylic acid

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 Jul - 03 Aug 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted in 1997

Deviations:

no

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted in 2020

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Version / remarks:

adopted in 1998

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Ministerium für Arbeit, Gesundheit und Soziales, Düsseldorf, Germany

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylic acid

EC Number:

700-093-4

Cas Number:

176969-34-9

Molecular formula:

C6H6F2N2O2

IUPAC Name:

3-(difluoromethyl)-1-methyl-1H-pyrazole-4-carboxylic acid

Method

Target gene:

His operon

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

Cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of male Sprague-Dawley rats treated with Aroclor 1254.
Prior to first use, each batch was checked for its metabolising capacity by using reference mutagens; appropriate activity was demonstrated.

Test concentrations with justification for top dose:

First and second experiment: 16, 50, 158, 500, 1581 and 5000 µg/plate with and without metabolic activation
Dose levels were routinely determined on the basis of a standard protocol: If not limited by solubility, 5000 µg/plate or 5 µL/plate were used as highest dose. The results of the first experiment were considered as a pre-test for toxicity. Concentrations of the second experiment were chosed based on the results of the first experiment.

Vehicle / solvent:

Vehicle/solvent used: DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

cumene hydroperoxide

mitomycin C

other: nitrofurantoin, -S9, 0.2 and 0.4 µg/plate in DMSO for TA100; 4-nitro-1,2-phenylene diamine, -S9, 10 and 20 µg/plate in DMSO for TA1537, 0.5 and 1 µg/plate in DMSO for TA98; 2-aminoanthracene, +S9, 3 and 6 µg/plate in DMSO for all strains

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration: triplicates
- Number of independent experiments : 2

METHOD OF APPLICATION: in agar (plate incorporation, Experiment 1) and preincubation (Experiment 2)

TREATMENT AND HARVEST SCHEDULE:
- Preincubation period: 20 min
- Exposure duration: 48 h

METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method: Cytotoxicity was determined as background growth inhibition, dose-dependent reduction in mutant count per plate and by titer determination.

Evaluation criteria:

A reproducible and dose-related increase in mutant counts of at least one strain is considered to be a positive result. For TA1535, TA100 and TA98 this increase should be about twice that of negative controls, whereas for TA1537, at least a threefold increase should be reached. For TA102 an increase of about 100 mutants should be reached. Otherwise, the result is evaluated as negative.

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium, other: TA98, TA100, TA102, TA1535 and TA1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

evident as a reduction in titer at ≥ 1581 µg/plate for all strains in the first experiment (plate incorporation test) and for strain TA 100 at 5000 µg/plate in the second experiment (pre-incubation test).

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS :
- Precipitation and time of the determination: There was no precipitation at any test item concentration in any experiment, neither with nor without S9 mix.

CYTOTOXICITY:
Cytotoxicity, evident as a reduction in titer was noted for all strains in the first experiment at ≥ 1581 µg/plate (plate incorporation test) and for strain TA 100 at 5000 µg/plate in the second experiment (pre-incubation test). There was no inhibition of the bacterial background lawn and no decrease in the number of spontaneous revertant colonies.

HISTORICAL CONTROL DATA: please refer to Table 3 under “Any other information on results incl. tables”

Any other information on results incl. tables

Table 1: Mean number of revertant colonies per plate observed in the first experiment (plate incorporation test)

Concentration (µg/plate)	TA 1535		TA 100		TA 1537		TA 98		TA 102
Metabolic activation	(-S9)	(+S9)	(-S9)	(+S9)	(-S9)	(+S9)	(-S9)	(+S9)	(-S9)	(+S9)
DMSO	8 ± 1	11 ± 2	110 ± 15	177 ± 22	6 ± 2	8 ± 1	23 ± 2	39 ± 2	294 ± 5	313 ± 64
16	7 ± 1	8 ± 1	101 ± 19	149 ± 23	6 ± 2	7 ± 1	22 ± 6	44 ± 7	301 ± 34	326 ± 76
50	8 ± 2	9 ± 1	96 ± 14	159 ± 29	5 ± 2	8 ± 0	22 ± 4	44 ± 2	269 ± 7	389 ± 28
158	8 ± 2	9 ± 2	102 ± 3	144 ± 13	5 ± 1	7 ± 2	21 ± 5	46 ± 9	279 ± 8	280 ± 90
500	7 ± 2	8 ± 2	115 ± 2	165 ± 21	4 ± 1	8 ± 1	18 ± 3	40 ± 4	205 ± 3	299 ± 51
1581	7 ± 1	8 ± 2	96 ± 15	114 ± 17	4 ± 1	8 ± 1	22 ± 5	43 ± 7	195 ± 60	333 ± 71
5000	8 ± 4	7 ± 2	98 ± 16	91 ± 11	5 ± 2	6 ± 1	21 ± 9	33 ± 5	206 ± 65	259 ± 78
Positive control^#	NaN3	2-AA	NF	2-AA	4-NPDA	2-AA	4-NPDA	2-AA	MMC	2-AA
a)	772 ± 28	113 ± 8	300 ± 59	1298 ± 138	35 ± 3	193 ± 38	109 ± 7	1731 ± 324	846 ± 18	582 ± 18
b)	917 ± 24	82 ± 2	398 ± 27	1396 ± 258	36 ± 8	48 ± 6	132 ± 6	1821 ± 210	1003 ± 117	1269 ± 170
#: NaN3: Sodium azide, NF: Nitrofurantoin, 4-NPDA: 4-Nitro-1,2-phenylene diamine, MMC: Mitomycin C, 2-AA: 2-Aminoanthracene a) lower concentration, 10 µg/plate for NaN3, 0.2 µg/plate for NF, 10µg/plate for 4-NDPA for TA1537, 0.5 µg/plate for TA98, 0.2 µg/plate for MMC, 3 µg/platefor 2-AA b) upper concentration, 20 µg/plate for NaN3, 0.4 µg/plate for NF, 20µg/plate for 4-NDPA for TA1537, 1 µg/plate for TA98, 0.4 µg/plate for MMC, 6 µg/plate for 2-AA

Table 2: Mean number of revertant colonies per plate observed in the second experiment (pre-incubation test)

Concentration (µg/plate)	TA 1535		TA 100		TA 1537		TA 98		plate
Metabolic activation	(-S9)	(+S9)	(-S9)	(+S9)	(-S9)	(+S9)	(-S9)	(+S9)	(-S9)	(+S9)
DMSO	9 ± 3	13 ± 3	134 ± 24	177 ± 21	7 ± 1	10 ± 3	16 ± 1	30 ± 4	249 ± 20	316 ± 11
16	9 ± 2	14 ± 3	146 ± 3	190 ± 20	7 ± 1	10 ± 4	17 ± 3	27 ± 4	251 ± 14	303 ± 15
50	8 ± 2	13 ± 2	131 ± 15	191 ± 7	7 ± 1	8 ± 1	17 ± 6	34 ± 3	251 ± 19	277 ± 47
158	11 ± 3	13 ± 2	146 ± 27	218 ± 21	7 ± 1	8 ± 1	18 ± 5	32 ± 2	216 ± 15	335 ± 18
500	8 ± 1	11 ± 4	122 ± 11	214 ± 31	6 ± 1	7 ± 1	20 ± 2	27 ± 4	250 ± 8	342 ± 27
1581	7 ± 2	8 ± 1	153 ± 5	188 ± 3	7 ± 2	8 ± 3	18 ± 1	31 ± 4	239 ± 9	335 ± 13
5000	8 ± 1	11 ± 2	132 ± 10	173 ± 10	7 ± 1	7 ± 1	19 ± 3	26 ± 6	254 ± 29	277 ± 36
Positive control^#	NaN3	2-AA	NF	2-AA	4-NPDA	2-AA	4-NPDA	2-AA	Cumene	2-AA
a)	1074 ± 15	112 ± 13	503 ± 48	2459 ± 451	34 ± 2	273 ± 36	72 ± 6	1538 ± 94	447 ± 8	539 ± 39
b)	850 ± 13	68 ± 21	927 ± 23	2553 ± 325	50 ± 4	183 ± 12	114 ± 17	2019 ± 117	454 ± 29	874 ± 35
#: NaN3: Sodium azide, NF: Nitrofurantoin, 4-NPDA: 4-Nitro-1,2-phenylene diamine, 2-AA: 2-Aminoanthracene a) lower concentration, 10 µg/plate for NaN3, 0.2 µg/plate for NF, 10µg/plate for 4-NDPA for TA1537, 0.5 µg/plate for TA98, 50 µg/plate for Cumene, 3 µg/platefor 2-AA b) upper concentration, 20 µg/platefor NaN3, 0.4 µg/plate for NF, 20µg/plate for 4-NDPA for TA1537, 1 µg/plate for TA98, 75µg/plate for Cumene, 6 µg/plate for 2-AA

Table 3: Historical control data generated in the testing laboratory from 1999 to 2008

1. Plate incorporation method Summary of Historical Negative and Positive Controls Using Mean Values Presented as Medians (Z) and Semi-Q Range (QR)

1999

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	8		82	22	7	1	21	3	185	28
DMSO	-	8	1	73	13	7	2	17	3	174	30
Ethanol	-	7	-	99	-	5	-	19	-	292	-
EGDE	-	12	-	90	-	6	-	19	-	298	-
DMF	-	7	-	70	-	5	-	22	-	198	-
Na-azid	-	642	79
NF	-			217	28
4-NPDA	-					105	15	137	20
Cumene	-									351	50
Water	+	9	2	102	31	7	2	26	7	273	22
DMSO	+	9	2	92	17	8	1	24	2	248	31
Ethanol	+	7	-	123	-	8	-	24	-	307	-
EGDE	+	10	*	115	-	8	-	26	-	385	-
DMF	+	10	-	98	-	8	-	24	-	266	-
2-AA	+	184	43	1290	304	221	57	1295	194	477	79

2000

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	9	1	81	11	7	2	25	6	196	32
DMSO	-	9	2	73	17	7	1	23	6	209	30
Ethanol	-	12	-	97	-	6	-	31	-	209	-
Na-azid	-	629	70
NF	-			226	35
4-NPDA	-					106	15	173	21
Cumene	-									352	43
Water	+	11	1	104	14	10	2	37	5	260	41
DMSO	+	10	2	94	20	8	2	32	7	270	34
Ethanol	+	12	-	106	-	13	-	42	-	230	-
2-AA	+	207	38	1468	223	252	88	1524	160	539	66

2001

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	9	3	100	22	6	1	39	4	223	42
DMSO	-	9	2	100	13	6	2	34	13	208	35
Acetone	-	8	-	113	-	8	-	38	-	233	-
Na-azid	-	633	76
NF	-			264	26
4-NPDA	-					94	13	172	19
Cumene	-									365	24
Water	+	11	2	119	22	9	2	50	6	283	30
DMSO	+	10	2	125	19	9	2	41	9	267	32
Acetone	+	12	-	129	-	10	-	45	-	317	-
2-AA	+	193	26	1614	97	270	78	1443	67	532	71

2002

Compound						Strain
and		TA 1535		TA 100		TA 1537		TA 98		TA 102
S9 Mix		Z	QR	Z	QR	Z	QR	Z	QR	Z	QR
Water	-	14	3	130	29	8	3	35	8	222	50
DMSO	-	14	5	136	15	7	2	31	9	216	35
EGDE	-	21	7	146	8	8	2	27	5	262	38
Na-azid	-	735	80
NF	-			323	30
4-NPDA	-					88	11	156	24
Cumene	-									556	92
Water	+	11	3	151	25	10	2	48	14	275	36
DMSO	+	12	4	151	19	8	2	39	12	270	24
EGDE	+	9	3	144	19	7	1	33	6	306	45
2-AA	+	158	30	1506	126	256	60	1283	139	554	64

2003

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	14	5	140	20	7	2	28	4	252	44
DMSO	-	14	3	144	13	7	1	24	5	227	28
EGDE	-	16	2	167	-	7	-	26	-	243	-
Ethanol	-	14	-	150	-	8	-	18	-	221	-
Acetone	-	15	-	134	-	9	-	42	-	279	-
Na-azid	-	679	90
NF	-			358	42
4-NPDA	-					91	12	157	18
MMC	-									530	63
Water	+	13	1	177	30	10	2	41	6	281	32
DMSO	+	11	2	169	22	9	2	34	6	294	32
EGDE	+	11	1	184	-	10	-	39	-	280	-
Ethanol	+	25	-	177	-	9	-	38	-	227	-
Acetone	+	10	-	172	-	12	-	31	*	311	-
2-AA	4-	170	28	1526	143	286	78	1247	138	721	114

2004

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	16	3	150	12	8	1	24	7	227	13
DMSO	-	16	4	135	15	7	1	26	9	199	14
EGDE	-	16	-	156	-	7	-	26	-	263	-
Ethanol	-	16	*	135	-	7	-	27	-	218	-
Na-azid	-	651	121
NF	-			339	28
4-NPDA	-					91	18	153	16
MMC	-									541	59
Water	+	13	2	158	20	11	2	42	11	255	20
DMSO	+	11	2	153	23	10	2	40	9	255	24
EGDE	+	12	-	191	-	8	-	48	-	301	-
Ethanol	+	20	-	170	-	10	-	40	-	251	-
2-AA	+	150	29	1436	116	259	57	1164	112	573	79

2005

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	19	7	162	22	7	1	25	5	223	20
DMSO	-	16	3	142	17	7	1	24	8	214	22
EGDE	-	19	3	152	33	7	1	29	7	254	27
Ethanol	-	11	-	138	-	7	-	21	-	238	-
DMF	-	10	-	125	-	6	-	24	-	212	-
Na-azid	-	652	66
NF	-			310	33
4-NPDA	-					89	12	149	17
MMC	-									518	38
Water	+	13		205	41	8	1	41	8	260	44
DMSO	+	11	2	178	22	8	2	44	8	284	30
EGDE	+	16	2	152	17	8	1	43	7	310	31
Ethanol	+	15	-	127	-	11	-	31	-	287	-
DMF	+	10	-	171	-	7	-	51	-	264	-
2-AA		168	28	1426	142	185	58	1157	162	718	99

2006

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	17	5	139	22	7	1	18	6	205	18
DMSO	-	15	3	129	18	7	1	25	7	214	24
EGDE	-	20	-	164	-	7	-	37	-	188	-
Na-azid	-	654	101
NF	-			303	37
4-NPDA	-					78	8	142	33
MMC										560	222
Water	+	11	2	144	28	8	1	28	4	268	26
DMSO	+	10	2	157	26	8	1	31	7	261	26
EGDE	+	10	-	178	-	8	-	37	-	279	-
2-AA	+	134	29	1359	126	181	71	1017	107	596	105

2007

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	16	7	143	19	7	1	24	3	213	13
DMSO	-	13	5	138	22	7	1	21	4	237	33
EGDE	-	14	-	119	-	7	-	27	-	186	-
Na-azid	-	681	127
NF	-			355	32
4-NPDA	-					87	9	152	15
MMC	-									610	72
Water	+	11	1	172	33	8	2	34	4	288	37
DMSO	+	11	2	174	28	9	1	29	5	285	45
EGDE	+	13	-	149	-	10	-	35	-	252	-
2-AA	+	153	33	1490	343	173	51	1079	204	568	105

2008

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	13	7	143	21	6	1	24	3	220	15
DMSO	-	13	3	149	10	8	1	21	5	250	23
Ethanol	-	14	-	145	-	6	-	29	-	285	-
Acetone	-	15	-	108	-	7	-	34	-	225	-
Na-azid^1	-	916	127
Na-azid^2		979	163
NF^1	-			382	30
NF^2				563	60
4-NPDA^1	-					87	10	149	16
4-NPDA^2						161	8	259	15
MMC^1	-									818	79
MMC^2										1000	35
Water	+	11	2	188	9	10	2	33	3	275	35
DMSO	+	11	1	168	20	10	2	35	6	309	27
Ethanol	+	13	-	142	-	7	-	41	-	359	-
Acetone	+	10	-	139	-	9	-	34	-	291	-
2-AA^1	+	112	22	2183	311	185	77	1619	259	726	74
2-AA^2		99	28	1691	374	143	60	1737	298	1393	205

1 low dose of respective positive control ^2high dose of respective positive control

2. Pre-incubation method

Summary of Historical Negative and Positive Controls Using Mean Values Presented as Medians (Z) and Semi-Q Range (QR)

1999

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	10	2	117	10	9	2	22	6	247	32
DMSO	-	7	2	72	12	6	1	15	3	223	20
Ethanol	-	7	-	90	-	7	-	24	-	294	-
EGDE	-	10	-	120	-	16	-	13	-	279	-
DMF	-	12	-	135	-	9	-	15	-	282	-
Na-azid	-	638	59
NF	-			258	29
4-NPDA	-					136	17	139	12
Cumene	-									525	54
Water	+	11	2	144	30	12	2	28	7	294	15
DMSO	+	8	2	88	13	8	1	23	3	298	22
Ethanol	+	11	-	96	*	6	-	28	-	312	-
EGDE	+	11		173	-	13	-	22	-	329	-
DMF	+	14	-	158	-	12	-	29	-	312	-
2-AA	+	149	32	1190	181	203	42	1117	149	467	53

2000

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	11	3	135	8	10	2	24	5	239	23
DMSO	-	8	2	80	16	8	2	21	6	237	20
Ethanol	-	9	-	103	-	11	-	44	-	228	-
Na-azid	-	620	72
NF	-			294	55
4-NPDA	-					136	14	176	22
Cumene	-									469	43
Water	+	12	2	148	15	12	2	31	4	297	28
DMSO	+	9	1	90	14	9	2	29	6	296	20
Ethanol	+	13	-	127	-	14	-	51	-	211	-
2-AA	+	178	36	1353	213	239	54	1349	160	466	39

2001

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	10	2	121	24	9	1	40	6	269	25
DMSO	-	8	2	107	16	7	1	32	6	229	24
Acetone	-	13	-	120	-	10	-	37	-	271	-
Na-azid	-	614	92
NF	-			403	44
4-NPDA	-					121	17	182	20
Cumene	-									429	48
Water	-	11	1	136	36	10	1	43	9	305	32
DMSO	+	10	2	123	20	9	2	40	6	281	27
Acetone	+	15	-	142	-	14	-	51	-	315	-
2-AA	+	196	28	1627	93	294	46	1442	120	470	47

2002

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	17	5	153	9	8	2	32	13	272	18
DMSO	-	12	3	145	13	8	2	29	6	255	32
EGDE	-	18	4	158	12	9	2	33	7	264	42
Na-azid	-	740	95
NF	-			447	31
4-NPDA	-					114	14	165	17
Cumene	-									480	35
Water	+	15	2	167	20	11	2	45	11	322	19
DMSO	+	11	2	155	16	9	1	38	9	314	38
EGDE	+	12	10	179	20	9	2	46	10	342	35
2-AA	+	170	22	1550	122	316	57	1234	149	509	34

2003

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	18	2	154	19	10	3	31	10	246	26
DMSO	-	15	3	137	18	8	1	20	5	236	24
EGDE	-	16	-	161	-	8	-	26	-	262	-
Ethanol	-	14	-	142	-	7	-	23	-	249	-
Acetone	-	20	-	134	-	12	-	33	-	340	-
Na-azid	-	664	88
NF	-			463	37
4-NPDA	-					120	18	164	17
Cumene	-									477	44
Water	+	13	5	173	32	9	1	39	11	303	63
DMSO	+	11	2	159	24	9	2	34	5	297	28
EGDE	+	15	-	213		12	-	52	-	288	-
Ethanol	+	21	-	181	-	8	-	39	-	308	-
Acetone	+	9	-	159	-	17	-	26	-	398	-
2-AA	+	187	28	1527	159	345	60	1255	144	613	94

2004

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	18	3	151	9	11	2	25	5	224	14
DMSO	-	15	3	134	10	8	1	24	6	209	19
EGDE	-	14	•	148	-	9	-	21	-	253	-
Ethanol	-	18	-	130	-	6	-	22	-	218	-
Na-azid	-	609	107
NF	-			460	56
4-NPDA	-					111	18	157	18
Cumene	-									470	34
Water	+	14	2	182	10	10	2	35	5	266	11
DMSO	+	12	2	156	24	10	2	36	7	251	25
EGDE	+	13	-	206	-	11	-	51	-	298	-
Ethanol	+	24	-	182	-	9	-	34	-	221	-
2-AA	+	163	14	1515	134	293	25	1145	120	512	56

2005

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	18	4	158	15	8	3	29	6	241	38
DMSO	-	17	4	144	18	7	2	27	8	228	19
EGDE	-	15	-	144	-	8	-	23	-	239	-
Ethanol	-	13	-	139	-	8	-	30	-	260	-
DMF	-	12	-	93	-	8	-	32	-	270	-
Na-azid	-	637	72
NF	-			451	44
4-NPDA	-					107	21	155	17
Cumene	-									428	39
Water	+	14	2	205	30	11	2	44	8	289	47
DMSO	+	11	3	183	31	9	2	44	7	289	33
EGDE	+	13	-	193	-	11		32	-	308	-
Ethanol	+	21	-	191	-	10	-	29	*	276	-
DMF	+	10	-	124	-	9	-	51	-	284	-
2-AA	+	161	18	1448	159	297	46	1142	97	607	99

2006

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	19	5	148	20	7	1	19	7	242	26
DMSO	-	15	3	130	14	7	1	19	4	217	23
EGDE	-	22	-	151	-	7	-	26	-	274	-
Na-azid	-	668	87
NF	-			444	33
4-NPDA	-					98	10	142	10
Cumene	-									443	26
Water	+	12	3	182	39	9	1	29	6	275	29
DMSO	+	10	2	142	28	9	1	30	5	273	29
EGDE	+	10	-	191	-	8	-	39	-	252	-
2-AA	+	142	25	1397	101	217	50	982	91	528	62

2007

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	16	6	157	20	8	2	26	4	256	37
DMSO	-	13	3	137	20	8	1	20	4	237	24
EGDE	-	14	-	160	-	8	-	27	-	284	-
Na-azid	-	683	118
NF	-			489	48
4-NPDA	-					107	12	156	13
Cumene	-									482	35
Water	+	11	2	214	29	9	1	33	4	322	35
DMSO	+	11	2	176	21	9	2	30	4	293	33
EGDE	+	13	-	205	-	8	-	34	-	306	-
2-AA	+	150	22	1473	231	203	54	1007	172	514	42

2008

Compound and S9 Mix		TA 1535 Z       QR		TA 100 Z       QR		Strain TA 1537 Z       QR		TA 98 Z       QR		TA 102 Z       QR
Water	-	17	6	151	16	9	1	21	6	249	21
DMSO	-	12	5	136	15	8	2	20	4	248	24
Ethanol		16	-	160	-	9	-	30	-	311	-
Acetone	-	16	-	145	-	10	-	25	-	216	-
EGDE	-	15	-	148	-	8	-	19	-	281	-
Na-azid^1	-	869	139
Na-azid^2		1063	82
NF^1	-			568	39
NF^2				884	60
4-NPDA^1	-					107	12	163	16
4-NPDA^2						211	15	271	13
Cumene^1	-									559	52
Cumene^2										595	41
Water	+	12	3	178	24	11	2	39	6	310	34
DMSO	+	13	2	164	28	11	2	33	5	309	28
Ethanol		15	-	159	-	13	-	40	*	353	-
Acetone	+	10	-	143	-	11	-	29	-	248	-
EGDE	+	15	*	182	-	14	-	36	-	322	-
2-AA^1	+	132	26	2244	283	245	67	1426	246	640	78
2-AA^2		108	10	1900	294	81	143^3	1461	118	1318	288

^1low dose of respective positive control ^2high dose of respective positive control ^3 high variations due to cytotoxicity

 

Applicant's summary and conclusion

Conclusions:

The test item was tested for bacterial mutagenicity in the Ames test according to OECD guideline 471. Under the conditions of the test, the test item was not mutagenic in S. typhimuirum strains TA98, TA100, TA102, TA1535 and TA1537 with and without metabolic activation.