Pentapotassium bis(peroxymonosulphate) bis(sulphate)

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:

2001-03-15 - 2001-10-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Pentapotassium bis(peroxymonosulphate) bis(sulphate)

EC Number:

274-778-7

EC Name:

Pentapotassium bis(peroxymonosulphate) bis(sulphate)

Cas Number:

70693-62-8

Molecular formula:

H3K5O18S4

IUPAC Name:

pentapotassium bis((hydroperoxysulfonyl)oxidanide) hydrogen sulfate sulfate

Test material form:

solid

Method

Target gene:

not indicated

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535

Additional strain / cell type characteristics:

other: histidine missense mutation (hisG46), deficient in a DNA repair system (uvrB), defective lipopolysaccharide coat on the wall (rfa)

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1537

Additional strain / cell type characteristics:

other: histidine frameshift mutation (hisC3076), defective in a DNA repair system and lipopolysaccharide coat

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 98

Additional strain / cell type characteristics:

other: histidine frameshift mutation (hisD3052), defective in a DNA repair system and lipopolysaccharide coat

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 100

Additional strain / cell type characteristics:

other: histidine missense mutation (hisG46), deficient in a DNA repair system (uvrB), defective lipopolysaccharide coat on the wall (rfa)

Species / strain 5

Species / strain / cell type:

E. coli WP2 uvr A pKM 101

Additional strain / cell type characteristics:

other: tryptophan dependent mutant

Metabolic activation:

with and without

Metabolic activation system:

S9 mix from Aroclor 1254 induced male Sprague-Dawley rats

Test concentrations with justification for top dose:

First test (range finding):
5000, 1500, 500, 150, 50, 15 and 5 µg/plate (with and without metabolic activation). Based on the results of the range-finding (cytotoxicity) test, the following concentrations were selected for the main test:

Second (main) test:
5000, 1500, 500, 150 and 50 µg/plate with metabolic activation
500, 150, 50, 15 and 5 µg/plate without metabolic activation

Vehicle / solvent:

not indicated

Controls

Untreated negative controls:

yes

Remarks:

water

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: Without metabolic activation: Sodium azide, 9 Aminoacridine, 2 Nitrofluorene, 2 (2 Furyl) 3 (5 nitro 2 furyl)acrylamide With metabolic activation: 2-Aminoanthracene, Benzo[a]pyrene

Details on test system and experimental conditions:

METHOD OF APPLICATION:
Aliquots of 0.1 mL of the test dilution (in water), positive or negative control, were placed in glass vessels.
0.5 mL S9 mix or 0.5 mL 0.1 M phosphate buffer (pH 7.4) was added, followed by 0.1 mL of a 10 hour bacterial culture and 2 mL of agar containing histidine (0.5 mM) and tryptophan (0.5 mM) at 45+/-2°C. The mixture was thoroughly shaken and overlaid onto previously prepared petri dishes containing 25 mL minimal agar. Three petri dishes were used for each concentration. All plates were incubated at 37°C for ca. 72 hours.

DURATION
- Preincubation period: In the second test:
Tubes, which contained mixtures of bacteria, buffer or S9 mix and test dilution, were incubated at 37°C for 30 min. with shaking before the addition of the agar overlay.
- Exposure duration: 72 hours

NUMBER OF CELLS EVALUATED: 10 E09

Evaluation criteria:

For a test to be considered valid the mean of the solvent/vehicle control revertant colony number for each strain should lie within the historical control range.
The positive control compounds must cause at least a doubling of mean revertant colony number over the negative control.
The mean number of revertant colonies for each treatment group was compared with those obtained for the solvent/vehicle control groups.

Statistics:

not indicated

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

5000 µg/plate with S9 mix; 500 µg/plate without S9 mix

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

5000 µg/plate with S9 mix; 500 µg/plate without S9 mix

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

5000 µg/plate with S9 mix; 500 µg/plate without S9 mix

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

5000 µg/plate with S9 mix; 500 µg/plate without S9 mix

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

5000 µg/plate with S9 mix; 1500 µg/plate without S9 mix;

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

GENOTOXICITY:
Please refer to tables 1 and 2, which are presented under "Remarks on results including tables and figures"
- without metabolic activation: No increase in the number of revertants/plate observed
- with metabolic activation: No increase in the number of revertants/plate observed

CYTOTOXICITY:
Toxicity was seen in all strains following exposure to KMPS triple salt at 5000 µg/plate in the presence of S9 mix, and at 1500 µg/plate (E. coli) or 500 µg/plate (S. typhimurium) in the absence of S9 mix.

Any other information on results incl. tables

Table 1: Results test 1 (plate-incorporation method)

Plate	S9 mix	Revertant colony mean counts
		TA98	TA100	TA1535	TA1537	WP2uvr/pKM101
S9mix, sterility check	+	0	0	0	0	0
Buffer, sterility check	-	0	0	0	0	0
KMPS triple salt, sterility check (5000 µg/plate)	-	0	0	0	0	0
KMPS triple salt, 5000 µg/plate	+	4±3*	8±7*	3±4*	1±1*	34±9
KMPS triple salt, 1500 µg/plate	+	39±4	129±7	15±2	15±2	130±12
KMPS triple salt, 500 µg/plate	+	40±5	133±9	17±6	20±7	124±9
KMPS triple salt, 150 µg/plate	+	38±6	145±15	22±2	20±4	150±19
KMPS triple salt, 50 µg/plate	+	39±5	135±14	17±5	20±6	163±22
KMPS triple salt, 15 µg/plate	+	45±10	140±11	20±5	22±3	141±12
KMPS triple salt, 5 µg/plate	+	48±5	143±10	19±2	18±2	143±12
Water, 0.1 mL/plate	+	45±3	148±3	21±3	22±2	162±8
KMPS triple salt, 5000 µg/plate	-	0±0*	0±0*	0±0*	0±0*	0±0
KMPS triple salt, 1500 µg/plate	-	0±0*	0±0*	0±0*	0±0*	12±9
KMPS triple salt, 500 µg/plate	-	1±1*	13±6*	2±2*	1±1*	129±12
KMPS triple salt, 150 µg/plate	-	37±4	121±4	13±4	14±3	117±1
KMPS triple salt, 50 µg/plate	-	37±4	139±8	15±4	11±3	114±3
KMPS triple salt, 15 µg/plate	-	36±6	132±13	17±3	11±2	131±12
KMPS triple salt, 5 µg/plate	-	37±2	115±16	20±4	14±2	125±15
Water, (0.1 ml/plate)	-	39±5	135±8	18±3	14±2	131±10
Benzo[a]pyrene, (5 µg/plate)	+	702±77	989±24	452±56	209±13	806±72
2-Nitrofluorene, (1 µg/plate)	-	319±25	622±58	291±46	382±20	527±21
10^-6 dilution of overnight culture, plated on nutrient agar	-	104±6	106±1	130±8	110±11	169±16

*   cytotoxicity

Table 2: Results test 2 (with pre‑incubation)

Plate	S9 mix	Revertant colony mean counts
		TA98	TA100	TA1535	TA1537	WP2uvr/pKM101
S9mix, sterility check	+	0	0	0	0	0
Buffer, sterility check	-	0	0	0	0	0
KMPS triple salt, sterility check (5000 µg/plate)	-	0	0	0	0	0
KMPS triple salt, 5000 µg/plate	+	13±4	41±5	6±2	2±1	21±2
KMPS triple salt, 1500 µg/plate	+	41±6	138±6	14±1	14±3	115±25
KMPS triple salt, 500 µg/plate	+	42±8	134±8	19±7	14±1	138±21
KMPS triple salt, 150 µg/plate	+	49±8	139±11	18±4	18±4	136±8
KMPS triple salt, 50 µg/plate	+	38±7	122±14	16±4	15±2	141±5
Water, 0.1 mL/plate	+	49±4	151±5	22±3	18±3	146±4
KMPS triple salt, 500 µg/plate	-	12±3	73±11	4±1	2±2	8±5
KMPS triple salt, 150 µg/plate	-	31±4	122±11	17±5	8±1	108±6
KMPS triple salt, 50 µg/plate	-	31±9	114±13	19±2	12±3	117±13
KMPS triple salt, 15 µg/plate	-	33±6	131±10	18±7	9±2	121±12
KMPS triple salt, 5 µg/plate	-	32±5	122±15	16±5	10±1	107±10
Water, (0.1 ml/plate)	-	33±6	123±6	19±1	13±3	120±13
Benzo[a]pyrene, (5 µg/plate)	+	778±71	1006±67	251±32	386±20	1461±396
2-Nitrofluorene, (1 µg/plate)	-	268±65	711±189	419±38	967±177	772±146
10^-6 dilution of overnight culture, plated on nutrient agar	-	116±3	176±10	131±16	120±8	180±14

Applicant's summary and conclusion

Conclusions:

The study and the conclusions which are drawn from it fulfill the quality criteria (validity, reliability, repeatability). Under the test conditions employed, KMPS triple salt showed no evidence of mutagenic activity in this bacterial system in the absence or presence of metabolic activation.

Executive summary:

Materials and methods

The mutagenic potential of KMPS triple salt was investigated in the Ames test using the histidine auxotroph  S. typhimurium  strains TA98, TA100, TA1535, TA1537, and a tryptophan dependent mutant of  E. coli, WP2uvrA/pKM101, all with and without metabolic activation by rat liver S9 mix from Aroclor 1254 induced male Sprague-Dawley rats. KMPS triple salt was tested at concentrations of 5000, 1500, 500, 150 and 50 µg/plate with metabolic activation and 500, 150, 50, 15 and 5 µg/plate without metabolic activity. The solvent used for the test substance was water.

 

Results and discussion

No substantial increases in revertant colony numbers over control counts were obtained with any of the tester strains following exposure to KMPS triple salt at any concentration in the presence or absence of S9 mix.

Toxicity was seen in all strains following exposure to KMPS triple salt at 5000 µg/plate in the presence of S9 mix, and at 1500 µg/plate (E. coli) or 500 µg/plate (S. typhimurium) in the absence of S9 mix.