Molybdenum nickel tetraoxide

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:

12 Apr - 05 Nov 2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study with acceptable restrictions (methodic deficiencies in cytotoxicity testing (determination of titre);

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

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:

Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht, Kaiser-Friedrich-Straße 7, 55116 Mainz, Germany

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Molybdenum nickel tetraoxide

EC Number:

238-034-5

EC Name:

Molybdenum nickel tetraoxide

Cas Number:

14177-55-0

Molecular formula:

MoO3*nNiO (with n in a range of 0.6 - 1)

IUPAC Name:

Reaction mass of molybdenum oxide and nickel oxide

Constituent 2

Reference substance name:

238-034-0

IUPAC Name:

238-034-0

Details on test material:

- Name of test material (as cited in study report): Molybdenum nickel tetraoxide
- Molecular formula: NiMoO4
- Physical state / appearance: light yellow powder
- Analytical purity: >99%
- Lot/batch No.: GRA20100315
- Expiration date of the lot/batch: 02/2012
- Storage condition of test material: in a closed vessel at room temperature (20 +/- 5 °C)

Method

Target gene:

His operon

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 97a, TA 98, TA 100 and TA 102

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 was produced from the livers of male Sprague-Dawley rats which were treated with 500 mg Aroclor 1254/kg bw intraperitoneally

Test concentrations with justification for top dose:

first experiment: 5010, 1508, 505, 151 and 51 µg/plate (nominal concentrations)
second experiment: 4979, 2529, 1252, 626, 354 µg/plate (nominal concentrations)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: deionised water
- Justification for choice of solvent/vehicle: water was chosen, because this solvent does not have any effects on the viability of the bacteria or the number of spontaneous revertants
The test item was not sufficiently soluble in the accepted solvents for the Ames-test. Therefore, suspensions of the test item in deionised water were prepared. All suspensions were constanly stirred before and during the test performance.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: -S9: TA 97a, TA 98, TA 102: 4-Nitro-1,2-phenylene diamine (NPD; 20 µg/plate); TA 100, TA 1535: sodium azide (NaN3; 1 µg/plate); +S9: TA 97a, TA 100, TA 102 and TA 1535: 2-Amino-anthracene (2-AA; 1 µg/plate); TA 98: Benzo-a-pyrene (BaP; 20 µg/plate)

Details on test system and experimental conditions:

First experiment:
METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Incubation time: 48 hours at 37 °C

Second experiment:
METHOD OF APPLICATION: pre-incubation method
DURATION
- Preincubation period: 20 minutes
- Incubation time: 48 hours at 37 °C

DETERMINATION OF CYTOTOXICITY
- Determination of titre: The titre was determined by dilution of the overnight culture using sodium chloride solution and placing 0.1 mL on maximal-soft agar. It should give a density of 10E9 cells/mL (at the least), correlating to 100 colonies/plate after dilution.
- Toxicity control: performed in experiment 1 only analogously to the titre control with the maximum dose of the test item (5010 µg/plate) with and without S9 mix on maximal soft agar (two replicates each). An additional toxicity control using 5 concentrations (53, 154, 497, 1499, 5003 µg/plate) was tested separately in the same manner.

Evaluation criteria:

A test item is considered to have mutagenic potential, if a significant, reproducible increase of revertant colonies per plate (increase factor ≥ 2) in at least one strain can be observed. A concentration-related increase over the range tested can also be taken as a signs of mutagenic activity.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA 97a, TA 98, TA 100, TA 102 and TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: Number of revertants of TA97, TA98 and TA100 was reduced compared to the control in the 1. experiment. The number of revertants of TA100, TA102 and TA1535 was reduced compared to the control in the 2. experiment. However, cytotoxicity testing was negative

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

Cytotoxicity of the test item was not detected. The background lawn was visible. As the revertants in the treatments of both experiments were decreased in comparison to the sponataneous revertants of the solvent control, an additional cytotoxicity test was performed. In this test no cytotoxicty was observed. Therefore it can be considered, that the test item is not cytotoxic under the conditions of the test.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Mean revertants of the first experiment

Strain		TA97a		TA98		TA100		TA102		TA1535
Induction		-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
H2O	Mean	159	121	17	17	138	128	188	185	14	13
	sd	21.5	10.2	0.5	2.4	22.3	47.2	19.4	60.0	1.3	3.9
DMSO	Mean	167	123	16	15	111	105	144	252	13	19
	sd	13.4	15.4	4.1	1.4	7.6	11.7	39.7	116.3	4.6	3.2
Pos.Contr.	Mean	510	694	562	121	658	607	1054	695	541	873
	sd	44	151	29	10	166	178	182	434	261	173
	f(I)	3.05	5.64	35.13	8.07	4.77	5.78	7.32	2.76	38.64	45.95
5010 µg/pl.	Mean	109	101	10	9	79	95	160	222	10	11
	sd	8	4	1	1	4	18	7	26	4	4
	f(I)	0.69	0.83	0.59	0.53	0.57	0.74	0.85	1.20	0.71	0.85
1508 µg/pl.	Mean	121	107	9	8	80	80	104	183	12	16
	sd	6	7	2	1	6	1	9	17	3	5
	f(I)	0.76	0.88	0.53	0.47	0.58	0.63	0.55	0.99	0.86	1.23
505 µg/pl.	Mean	107	108	9	10	98	99	167	266	17	14
	sd	11	7	2	1	23	8	23	24	5	2
	f(I)	0.67	0.89	0.53	0.59	0.71	0.77	0.89	1.44	1.21	1.08
151 µg/pl.	Mean	112	105	6	7	98	92	227	263	17	14
	sd	2	13	1	2	27	19	72	39	5	5
	f(I)	0.70	0.87	0.35	0.41	0.71	0.72	1.21	1.42	1.21	1.08
51 µg/pl.	Mean	115	111	9	8	81	98	184	175	10	23
	sd	6	5	1	1	5	15	52	35	1	8
	f(I)	0.72	0.92	0.53	0.47	0.59	0.77	0.98	0.95	0.71	1.77

sd = standard deviation

f(I) = increase factor

 

 

Table 2: Mean revertants of the second experiment

Strain		TA97a		TA98		TA100		TA102		TA1535
Induction		-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
H2O	Mean	135	122	9	11	105	127	162	236	10	17
	sd	24.7	12.4	5.0	3.5	2.2	29.9	46.5	20.3	1.3	2.9
DMSO	Mean	130	155	10	12	113	115	158	193	11	18
	sd	25.2	34.7	2.5	3.0	4.5	20.8	37.0	44.9	1.3	7.8
Pos.Contr.	Mean	603	680	640	341	261	379	510	619	317	374
	sd	142	152	64	20	64	62	161	213	181	182
	f(I)	4.64	4.39	64.00	28.42	2.49	3.30	3.23	3.21	31.70	20.78
4979 µg/pl.	Mean	130	139	10	10	143	83	161	158	11	10
	sd	12	6	1	1	37	14	10	31	2	3
	f(I)	0.96	1.14	1.11	0.91	1.36	0.65	0.99	0.67	1.10	0.59
2592 µg/pl.	Mean	143	116	10	10	117	107	131	160	13	10
	sd	6	17	1	4	33	14	12	19	2	2
	f(I)	1.06	0.95	1.11	0.91	1.11	0.84	0.81	0.68	1.30	0.59
1252 µg/pl.	Mean	124	133	11	12	88	121	170	135	13	13
	sd	11	12	3	2	9	33	14	31	3	5
	f(I)	0.92	1.09	1.22	1.09	0.84	0.95	1.05	0.57	1.30	0.76
626 µg/pl.	Mean	122	109	12	14	102	118	155	173	9	14
	sd	13	3	4	4	15	6	25	8	2	1
	f(I)	0.90	0.89	1.33	1.27	0.97	0.93	0.96	0.73	0.90	0.82
354 µg/pl.	Mean	105	112	11	14	105	93	127	209	8	11
	sd	9	12	4	3	7	18	8	21	3	2
	f(I)	0.78	0.92	1.22	1.27	1.00	0.73	0.78	0.89	0.80	0.65

sd = standard deviation

f(I) = increase factor

 

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with and without metabolic activation