Manganese bis(phosphinate)

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from peer reviewed publication

Data source

Reference

Reference 1

Reference Type:

publication

Title:

Gene mutation toxicity study of the test chemical

Author:

Prival et al

Year:

1991

Bibliographic source:

Mutation Research

Reference 2

Reference Type:

publication

Title:

Gene mutation toxicity study of the test chemical

Author:

Mortelmans et al

Year:

1980

Bibliographic source:

Department of Health, Education and Welfare, Food and Drug Administration

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Manganese bis(phosphinate)

EC Number:

233-143-4

EC Name:

Manganese bis(phosphinate)

Cas Number:

10043-84-2

Molecular formula:

H3O2P1/2Mn

IUPAC Name:

manganese bis(phosphinate)

Details on test material:

- Name of test material :manganese(2+) bis(phosphinate)
- Molecular formula : H3O2P1/2Mn
- Molecular weight : 184.9136 g/mol
- Smiles notation : [Mn+2].[O-]P=O.[O-]P=O
- InChl : 1S/Mn.2H3O2P/c;2*1-3-2/h;2*3H2,(H,1,2)/q+2;;/p-2
- Substance type: Inorganic
- Physical state: Solid

Method

Target gene:

Histidine for Salmonella and tryptophan for E. coli

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium, other: TA98, TA100, TA1535, TA1537 and TA1538

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

other: Histidine auxotroph

Species / strain 2

Species / strain / cell type:

E. coli WP2

Details on mammalian cell type (if applicable):

No data

Additional strain / cell type characteristics:

other: Tryptophan auxotroph

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

10% Aroclor 1254-induced liver S9 from male Sprague-Dawley rats.

Test concentrations with justification for top dose:

0, 0.033, 0.10, 0.33, 1.0, 3.3 and 10 mg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: 0.067 M sodium phosphate buffer
- Justification for choice of solvent/vehicle: The chemical was soluble in sodium phosphate buffer

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Sodium phosphate buffer

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

other: AF-2 (furylfuramide) or N-methyl-N'-nitro-N-nitrosoguanidine (E. coli WP2;without S9), -Anthramine (All strains; with S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: No data
- Exposure duration: No data
- Expression time (cells in growth medium): No data
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: All platings were performed in duplicate and all tests were repeated.

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data

OTHER: No data

Rationale for test conditions:

No data

Evaluation criteria:

Test results were considered valid only if the positive control compounds induced increases in mutant counts to at least twice background.

Statistics:

None--straight counting of colonies on plates.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium, other: TA98, TA100, TA1535, TA1537 and TA1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data
- Effects of osmolality: No data
- Evaporation from medium: No data
- Water solubility: No data
- Precipitation: No data
- Definition of acceptable cells for analysis: No data
- Other confounding effects: No data

RANGE-FINDING/SCREENING STUDIES: The test compound was first tested in a range finding assay at dose levels between 0.3 and 10000 µg/plate

CYTOKINESIS BLOCK (if used)
- Distribution of mono-, bi- and multi-nucleated cells: No data

NUMBER OF CELLS WITH MICRONUCLEI
- Number of cells for each treated and control culture: No data
- Indication whether binucleate or mononucleate where appropriate: No data

HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%)
- Positive historical control data: No data
- Negative (solvent/vehicle) historical control data: No data

ADDITIONAL INFORMATION ON CYTOTOXICITY:
- Measurement of cytotoxicity used: No data
- Other observations when applicable: No data

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table: In vitro assay results

Compound	Metabolic activation	Dose (µg/plate)	Histidine revertants/plate TA1535
			Experiment 1			Experiment 2
					Average			Average
Negative control	-		16	18	20	18	15	24
	-		26	21		34	29
	+		16	15	15	14	13	10
	+		14	14		6	8
Positive control
Sodium azide	-	1.0	91	103	97	382	435	409
2-Anthramine	-	2.5	25	19	22	24	20	22
	+	2.5	304	281	293	322	325	324
Manganese hypophosphite	-	33.0	15	20	18	17	26	22
	-	100.0	27	24	26	19	25	22
	-	333.3	18	26	22	18	14	16
	-	1000.0	17	31	24	21	24	23
	-	3333.3	25	24	25	20	21	21
	-	10000.0	19	13	16	20	19	20
	+	33.0	17	18	18	9	10	10
	+	100.0	20	18	19	10	12	11
	+	333.3	18	15	17	14	10	12
	+	1000.0	13	14	14	10	13	12
	+	3333.3	19	18	19	7	12	10
	+	10000.0	21	11	16	14	7	11

 

Compound	Metabolic activation	Dose (µg/plate)	Histidine revertants/plate TA1537
			Experiment 1			Experiment 2
					Average			Average
Negative control	-		8	7	7	10	6	6
	-		6	8		2	4
	+		6	7	10	9	15	11
	+		12	16		10	11
Positive control
9- Aminoacridine	-	50.0	201	195	198	306	269	288
2-Anthramine	-	2.5	20	6	13	7	8	8
	+	2.5	190	220	205	89	77	83
Manganese hypophosphite	-	33.0	8	7	8	6	7	7
	-	100.0	2	3	3	6	11	9
	-	333.3	6	8	7	10	14	12
	-	1000.0	5	8	7	7	9	8
	-	3333.3	8	10	9	6	7	7
	-	10000.0	7	5	6	9	8	9
	+	33.0	8	8	8	12	15	14
	+	100.0	8	15	12	20	17	19
	+	333.3	14	7	11	8	11	10
	+	1000.0	5	9	7	18	9	14
	+	3333.3	9	8	9	7	10	9
	+	10000.0	14	7	11	15	11	13

 

Compound	Metabolic activation	Dose (µg/plate)	Histidine revertants/plate TA1538
			Experiment 1			Experiment 2			Experiment 3
					Avrg			Avrg			Avrg
Negative control	-		28	28	29	19	14	16	25	18	22
	-		36	26		15	16
	+					16	22	1	19	25	20
	+					14	15		18	16
Positive control
2-Nitrofluorene	-	5.0	483	530	507	897	977	937	796	813	805
2-Anthramine	-	1.0	17	17	17	11	14	13	12	13	13
	+	1.0				131	197	164	417	437	427
Manganese hypophosphite	-	33.0	16	8	12	10	9	10
	-	100.0	15	C*	15	10	17	14
	-	333.3	17	19	18	13	13	13
	-	1000.0	30	26	28	8	9	8
	-	3333.3	25	21	23	10	9	10
	-	10000.0	32	20	26	13	7	10
	+	33.0	C	C		8	7	8	21	16	19
	+	100.0	C	C		12	23	18	10	28	19
	+	333.3	C	C		26	17	22	20	18	19
	+	1000.0	C	C		9	18	14	16	27	22
	+	3333.3	C	C		20	13	17	22	13	18
	+	10000.0	C	C		11	18	15	22	21	22

*C: contaminated

 

Compound	Metabolic activation	Dose (µg/plate)	Histidine revertants/plate TA98
			Experiment 1			Experiment 2
					Average			Average
Negative control	-		68	48	51	40	33	38
	-		48	40		45	33
	+		43	31	41	42	60	48
	+		38	51		40	51
Positive control
2- Nitrofluorene	-	5.0	356	452	404	418	524	471
2-Anthramine	-	1.0	39	25	32	48	35	42
	+	1.0	178	219	198	212	206	209
Manganese hypophosphite	-	33.0	21	40	31	35	32	34
	-	100.0	30	34	32	30	35	33
	-	333.3	30	38	34	45	51	48
	-	1000.0	19	40	30	35	41	38
	-	3333.3	27	45	36	39	41	40
	-	10000.0	33	52	43	38	24	31
	+	33.0	21	23	22	37	40	39
	+	100.0	27	43	35	47	48	48
	+	333.3	22	37	30	42	67	55
	+	1000.0	33	28	31	40	51	46
	+	3333.3	43	18	31	51	49	50
	+	10000.0	C*	C	C	39	43	41

*C: contaminated

 

Compound	Metabolic activation	Dose (µg/plate)	Histidine revertants/plate TA100
			Experiment 1			Experiment 2
					Average			Average
Negative control	-		99	100	95	93	93	97
	-		100	81		84	117
	+		122	96	109	104	91	95
	+		104	112		101	82
Positive control
Sodium azide	-	1.0	499	441	470	354	356	355
2-Anthramine	-	1.0	108	109	109	102	120	111
	+	1.0	349	352	351	371	405	388
Manganese hypophosphite	-	0.3	116	98	107	35	32	34
	-	3.3	108	98	98	30	35	33
	-	33.3	92	101	97	102	105	104
	-	100.0	113	111	112	112	97	105
	-	333.3	136	100	118	99	104	102
	-	1000.0	91	109	100	105	98	102
	-	3333.3	93	93	93	126	125	126
	-	10000.0	108	108	108	108	100	104
	+	0.3	122	88	105
	+	3.3	101	97	99
	+	33.3	90	105	98	104	102	103
	+	100.0	105	101	103	110	115	113
	+	333.3	92	92	92	86	120	103
	+	1000.0	101	72	87	109	104	107
	+	3333.3	87	81	84	115	101	108
	+	10000.0	90	84	87	114	125	120

 

Compound	Metabolic activation	Dose (µg/plate)	Histidine revertants/plate E. coli WP2
			Experiment 1			Experiment 2
					Average			Average
Negative control	-		20	21	26	41	49	48
	-		32	31		50	50
	+		45	49	47	50	48	52
	+		53	42		72	39
Positive control
AF-2	-	0.1	516	574	545	146	152	149
2-Anthramine	-	10.0	27	24	26	18	30	24
	+	10.0	605	662	634	263	285	274
Manganese hypophosphite	-	33.3	29	24	27	30	39	35
	-	100.0	32	27	30	42	38	40
	-	333.3	33	32	33	39	45	42
	-	1000.0	21	26	24	61	40	51
	-	3333.3	41	37	39	43	48	46
	-	10000.0	37	32	35	52	52	52
	+	33.3	41	34	38	51	60	56
	+	100.0	42	48	45	64	64	64
	+	333.3	52	49	51	52	52	52
	+	1000.0	39	51	45	26	62	44
	+	3333.3	33	68	51	65	77	71
	+	10000.0	49	65	57	89	82	86

 

Applicant's summary and conclusion

Conclusions:

The test chemical did not induce mutation in the Salmonella typhimurium strain TA98, TA100, TA1535, TA1537 and TA1538 and Escherichia coli strain WP2 with and without rat liver S9 mix and hence the chemical is negative for gene mutation in vitro.

Executive summary:

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed as per the plate incorporation protocol and the test chemical dissolved in 0.067M sodium phosphate buffer and was used at dose levels of 0, 0.033, 0.10, 0.33, 1.0, 3.3 and 10 mg per plate. Concurrent solvent and positive controls were run with the test chemical. Test results were considered valid only if the positive control compounds induced increases in mutant counts to at least twice background. The test chemical did not induce mutation in the Salmonella typhimurium strain TA98, TA100, TA1535, TA1537 and TA1538 and Escherichia coli strain WP2 with and without rat liver S9 mix and hence the chemical is negative for gene mutation in vitro.