Aluminum magnesium vanadium oxide

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 - 18 Oct 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP - Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

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, Mainz, Germany

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

divanadium(5+) dialuminium(3+) magnesium(2+) nonaoxidandiide

EC Number:

692-448-4

Cas Number:

170621-28-0

Molecular formula:

Mg2xAl2xVyOz

IUPAC Name:

divanadium(5+) dialuminium(3+) magnesium(2+) nonaoxidandiide

Test material form:

solid: particulate/powder

Details on test material:

light yellow

Method

Target gene:

his operon (S. thyphimurium)

Species / strain

Species / strain / cell type:

other: TA 1535, TA97a, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

co-factor supplemented post-mitochondrial fraction (S9 mix) (Trinova Biochem, Gießen, Germany), prepared from the livers of male Sprague Dawley rats intraperitoneally treated with 500 mg/kg bw Aroclor 1254

Test concentrations with justification for top dose:

First experiment: 51, 150, 499, 1502 and 5002 µg/plate with and without metabolic activation
Second experiment: 315, 625, 1253, 2503 and 5001 µg/plate with and without metabolic activation

Vehicle / solvent:

- Vehicle/solvent used: sterile, demineralised water (0.1 mL for the plate incorporation and the preincubation treatment)
- Justification for choice of solvent/vehicle: water was chosen as vehicle, since the test substance was not soluble in any other of the recommended vehicles and no effects on viability.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-1,2-phenylene diamine (20 µg/plate, -S9, TA 97a, TA98 and TA 102); sodium azide (1 µg/plate, -S9, TA100 and TA 1535); 2-amino-anthracene (1 µg/plate, +S9, TA 97a, TA100, TA 102 and TA 1535); benzo-a-pyrene (20 µg/plate, +S9, TA98)

Details on test system and experimental conditions:

METHOD OF APPLICATION 1: in agar (plate incorporation) for first experiment

DURATION
- Exposure duration: 48 h

METHOD OF APPLICATION 2: preincubation followed by plate incorporation for the second experiment

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

NUMBER OF REPLICATIONS: quadruplicate each in both experiments

DETERMINATION OF CYTOTOXICITY
- Method: determination of titre (cytotoxicity test) and observation of background lawn (plate incorporation and preincubation experiment)

Evaluation criteria:

The test substance was considered to have mutagenic potential, if a significant, reproducible increase in revertant colonies per plate (increase factor ≥ 2) in at least one of the test strains was observed. A concentration-dependent increase over a range of test concentrations was considered to be a sign of mutagenic activity.

Statistics:

Mean values and standard deviations of revertant colonies from each test group were calculated.

Results and discussion

Test results

Key result

Species / strain:

other: TA 1535, TA97a, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: the test substance was only suspendable in water. Thus, it had to be stirred during application. All concentrations were freshly prepared.

COMPARISON WITH HISTORICAL CONTROL DATA:
The mean numbers of spontaneous revertants in the solvent/vehicle controls of the strains used were all within the normal historical ranges.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
In a preliminary study using the plate incorporation method, each strain was incubated with 1511 and 5004 µg/plate of the test substance for 48 h. Two replicates for each concentration were performed both in the presence or absence of metabolic activation. For the determination of toxicity, the titre (colonies/plate) was determined in control and treated cultures. In control culture, the titre should give a number of at least 1E+9 cells/mL, correlating to 100 colonies/plate after dilution. A substance is considered non-toxic, if the quotient control titre/treatment titre is below 2. This criterion was fulfilled after treatment with 1511 and 5004 µg/plate of the test substance. In the main study, no signs of toxicity were observed in any strain and at any concentration tested in both experiments.

Any other information on results incl. tables

Table1. Test results of first experiment (plate incorporation)

Aluminium magnesium vanadium oxide: Bacterial Reverse Mutation Assay, mean revertant colonies/plate (n=4 ± SD)
EXPERIMENT I (plate incorporation)
S9-Mix	Without
Concentration (per plate)	TA 97a	TA98	TA100	TA102	TA1535
SC (water)	110 ± 3	16 ± 2	125 ± 14	159 ± 12	14 ± 4
SC (DMSO)	107 ± 5	12 ± 3	144 ± 33	155 ± 20	15 ± 3
Test item
5002 µg	107 ± 8	15 ± 2	101 ± 7	145 ± 8	11 ± 4
1502 µg	110 ± 8	15 ± 2	94 ± 4	149 ± 6	14 ± 1
499 µg	100 ± 10	14 ± 1	92 ± 5	148 ± 5	11 ± 4
150 µg	106 ± 6	15 ± 4	105 ± 7	145 ± 5	9 ± 2
51 µg	106 ± 4	14 ± 3	96 ± 10	143 ± 7	9 ± 1
PC
4NOPD (20 µg)	703 ± 26	275 ± 37	-	622 ± 64	-
NaN3 (1 µg)	-	-	645 ± 69	-	254 ± 25
S9-Mix	With
Concentration (per plate)	TA 97a	TA98	TA100	TA102	TA1535
SC (water)	101 ± 7	16 ± 2	99 ± 7	195 ± 14	17 ± 1
SC (DMSO)	102 ± 3	13 ± 2	104 ± 12	186 ± 23	16 ± 0
Test item
5002 µg	118 ± 7	14 ± 2	99 ± 4	149 ± 7	12 ± 2
1502 µg	101 ± 2	16 ± 3	100 ± 8	151 ± 14	12 ± 2
499 µg	108 ± 6	14 ± 1	92 ± 3	153 ± 4	10 ± 2
150 µg	114 ± 1	14 ± 2	101 ± 8	148 ± 6	8 ± 2
51 µg	117 ± 4	14 ± 1	104 ± 12	149 ± 14	11 ± 14
PC
2AA (1 µg)	490 ± 113	-	548 ± 50	511 ± 90	143 ± 3
BP (20 µg)	-	251 ± 35	-	-	-
SC = Solvent control; PC = Positive control substances; SD = standard deviation; NaN3 = sodium azide; 4NOPD = 4-nitro-o-phenylene-diamine; 2AA = 2-aminoanthracene; BP = benzo-a-pyrene

Table2. Test results of second experiment (pre-incubation)

Aluminium magnesium vanadium oxide: Bacterial Reverse Mutation Assay, mean revertant colonies/plate (n=4 ± SD)
EXPERIMENT II (pre-incubation)
S9-Mix	Without
Concentration (per plate)	TA 97a	TA98	TA100	TA102	TA1535
SC (water)	111 ± 3	16 ± 6	81 ± 9	202 ± 14	23 ± 5
SC (DMSO)	115 ± 4	15 ± 4	130 ± 14	197 ± 27	21 ± 8
Test item
5001 µg	109 ± 4	10 ± 0	115 ± 6	209 ± 17	19 ± 1
2503 µg	102 ± 13	13 ± 2	133 ± 58	196 ± 12	14 ± 3
1253 µg	114 ± 11	15 ± 4	122 ± 13	181 ± 7	16 ± 6
625 µg	111 ± 5	10 ± 0	119 ± 15	203 ± 13	21 ± 4
315 µg	133 ± 19	10 ± 0	111 ± 4	199 ± 15	16 ± 4
PC
4NOPD (20 µg)	479 ± 70	229 ± 23	-	499 ± 31	-
NaN3 (1 µg)	-	-	338 ± 52	-	254 ± 66
S9-Mix	With
Concentration (per plate)	TA 97a	TA98	TA100	TA102	TA1535
SC (water)	118 ± 21	20 ± 8	96 ± 16	210 ± 8	19 ± 3
SC (DMSO)	115 ± 15	18 ± 4	96 ± 9	210 ± 21	20 ± 7
Test item
5001 µg	101 ± 13	11 ± 2	111 ± 3	192 ± 11	18 ± 3
2503 µg	113 ± 6	15 ± 2	142 ± 19	206 ± 12	18 ± 6
1253 µg	113 ± 6	13 ± 4	123 ± 3	199 ± 17	20 ± 1
625 µg	112 ± 8	10 ± 0	119 ± 18	200 ± 15	21 ± 5
315 µg	114 ± 6	13 ± 3	113 ± 8	182 ± 16	19 ± 1
PC
2AA (1 µg)	439 ± 47	-	987 ± 169	485 ± 100	145 ± 19
BP (20 µg)	-	324 ± 75	-	-	-
SC = Solvent control; PC = Positive control substances; SD = standard deviation; NaN3 = sodium azide; 4NOPD = 4-nitro-o-phenylene-diamine; 2AA = 2-aminoanthracene; BP = benzo-a-pyrene

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative