[[(phosphonomethyl)imino]bis[ethylenenitrilobis(methylene)]]tetrakisphosphonic acid, potassium salt

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

02-Jul-2001 to 26-Jul-2001

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was conducted according to the appropriate OECD test guideline, and in compliance with GLP. The original study was considered reliability 1. Read-across to the registered substance is considered scientifically justified and is reliability 2.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

(only 2-AA used as positive control +S9)

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Japanese Guidelines on Industrial Chemicals

GLP compliance:

yes

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

DTPMP-xNa

IUPAC Name:

DTPMP-xNa

Constituent 2

Reference substance name:

[[(phosphonomethyl)imino]bis[(ethylenenitrilo)bis(methylene)]]tetrakisphosphonic acid, sodium salt

EC Number:

244-751-4

EC Name:

[[(phosphonomethyl)imino]bis[(ethylenenitrilo)bis(methylene)]]tetrakisphosphonic acid, sodium salt

Cas Number:

22042-96-2

IUPAC Name:

sodium hydrogen [10,10-dihydroxy-10-oxido-2,5,8-tris(phosphonomethyl)-2,5,8-triaza-10-phosphadec-1-yl]phosphonate

Method

Target gene:

histidine (Salmonella typhimurium)
tryptophan (Escherichia coli)

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:

phenobarbital- and 5,6-benzoflavone-induced rat liver S9

Test concentrations with justification for top dose:

Test 1-1: 78-5000 µg mixed sodium salts/plate -S9, 156-5000 µg/plate +S9
Test 1-2: 20-5000 µg mixed sodium salts/plate -S9 for S typhimurium
Test 2: 20-5000 µg mixed sodium salts/plate -S9 for S typhimurium, 78-5000 µg mixed sodium salts/plate -S9 for E coli, 156-5000 µg mixed sodium salts/plate +S9
20-5000ug/plate -S9; 156-5000ug/plate +S9

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: solubility

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

without S9; TA98, 0.1 µg/plate; TA100, 0.01 µg/plate

Positive control substance:

furylfuramide

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

without S9 ; TA1535, 0.5 µg/plate

Positive control substance:

sodium azide

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

without S9; TA1537, 80 µg/plate

Positive control substance:

9-aminoacridine

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

without S9, WP2 uvrA, 2 µg/plate

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

with S9; TA98, 0.5 µg/plate; TA100, 1 µg/plate; TA1535, TA1537, 2 µg/plate; WP2 uvrA, 10 µg/plate

Positive control substance:

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

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

NUMBER OF REPLICATIONS: triplicate plates

DETERMINATION OF CYTOTOXICITY
- Method: other: growth inhibition

Evaluation criteria:

Positive: mean number of revertant colonies more than double the negative control value and significant concentration-dependent Increase

Statistics:

No statistical analysis performed

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

Remarks:

>= 625 µg mixed sodium salts/plate -S9

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

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

Remarks:

>=1250 µg mixed sodium salts/plate -S9

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

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: soluble in water
- Precipitation: no precipitation observed
- Other confounding effects: no data

RANGE-FINDING/SCREENING STUDIES:
- Concentrations of 5-5000 µg/plate
- No growth inhibition in any strain
- Bacteriostatics observed at 1250 µg/plate -S9 in all strains
- No precipitation

COMPARISON WITH HISTORICAL CONTROL DATA:
- Negative/solvent control: within the range of historical data
- Positive controls: within the range of historical data

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

All treated cultures had less than 2-fold increase in mutant frequency and no evidence of a dose-response.  All positive controls gave a greater than 2-fold increase in mutant numbers.

Table 1 Experiment 1 Mean number of revertants (average of 3 plates)

 

Concentration (µg/plate)	+/- S9 mix	TA 100	TA 1535	WP2uvrA	TA 98	TA 1537
Control	-	102	10	23	14	7
78	-	102	8	21	16	9
156	-	88	7	18	16	6
313	-	71	5	18	8	1
625	-	0	0	7	0	0
1250	-	0	0	0	0	0
2500	-	0	0	0	0	0
5000	-	0	0	0	0	0
Positive control	-	528	422	631	559	194
Control	+	111	11	25	24	6
156	+	133	11	30	24	10
313	+	148	11	21	22	6
625	+	140	9	23	17	6
1250	+	122	6	21	11	3
2500	+	108	4	15	9	3
5000	+	98	5	9	8	2
Positive control	+	501	260	648	446	154
Mean number or revertants 16-19.07.2001

Table 2 Experiment 2 Mean number of revertants (average of 3 plates)

Concentration (µg/plate)	+/- S9 mix	TA 100	TA 1535	WP2uvrA	TA 98	TA 1537
Control	-	104	11	-	16	7
20	-	101	6	-	16	8
39	-	94	8	-	12	8
78	-	106	8	-	14	8
156	-	97	9	-	14	5
313	-	81	6	-	14	4
1250	-	0	0	-	0	0
5000	-	0	0	-	0	0
Positive control	-	522	411	-	604	211

 

Table 3 Experiment 3 Mean number of revertants (average of 3 plates)

Concentration (µg/plate)	+/- S9 mix	TA 100	TA 1535	WP2uvrA	TA 98	TA 1537
Control	-	99	8	20	21	7
20	-	105	10	-	18	7
39	-	107	11	-	19	6
78	-	110	10	22	19	6
156	-	103	11	14	17	7
313	-	86	8	16	15	4
625	-	-	-	13	-	-
1250	-	0	0	0	0	0
2500	-	-	-	0	-	-
5000	-	0	0	0	0	0
Positive control	-	510	419	655	601	183
Control	+	127	10	21	23	8
156	+	130	9	29	26	9
313	+	132	12	17	19	7
625	+	127	12	22	18	6
1250	+	114	8	15	14	7
2500	+	119	6	13	6	3
5000	+	103	6	12	3	2
Positive control	+	679	304	758	517	152

 

Applicant's summary and conclusion

Conclusions:

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

DTPMP-xNa has been tested in a reliable bacterial mutagenicity assay, conducted according to OECD guideline 471 and in compliance with GLP. No increase in the number of revertants was observed in any strain with or without metabolic activation, in either the initial or the repeat assay. It is concluded that the substance is negative for mutagenicity to bacteria under the conditions of the test.