Potassium 3-sulphopropyl methacrylate

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:

25 January - 31 January 1983

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP - guideline study

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1983

Report date:

1983

Reference 2

Reference Type:

other: Amendment to study report project 015041

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD Guideline Draft No. 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

(strains: Salmonella typhimurium TA1535, TA 1537, TA 98, TA100 and TA 1538 were used)

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EU Method B.14 (Draft) , (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

yes

Remarks:

(strains: Salmonella typhimurium TA1535, TA 1537, TA 98, TA100 and TA 1538 were used)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Potassium 3-sulphonatopropyl acrylate

EC Number:

250-465-0

EC Name:

Potassium 3-sulphonatopropyl acrylate

Cas Number:

31098-20-1

Molecular formula:

C6H10O5S.K

IUPAC Name:

potassium 3-(acryloyloxy)propane-1-sulfonate

Details on test material:

- Name of test material (as cited in study report): 3-Sulfopropyl-methacrylsauereester , SPM
- Substance type: white crystal
- Analytical purity: 98%
- Lot/batch No.: 31098-21-2
- Storage condition of test material: at room temperature , in the dark
- Other: To avoid any light effects on the test compound, all experimentation was performed under yellow light.

Method

Target gene:

his-

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:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

Liver microsomal fraction (S9 mix), prepared from the livers of male Wistar rats treated with Aroclor 1254

Test concentrations with justification for top dose:

1.58, 5, 15.8, 50, 158, 500, 1580 and 5000 µg/plate with and without metabolic activation

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: distilled water
- On the day of the experiment, the test material was dissolved in distilled water at the highest investigated dose. The other doses were dilutions from this stock solution with the solvent in half-log intervals.

Controls

Untreated negative controls:

other: solvent control (distilled water)

Negative solvent / vehicle controls:

yes

Remarks:

distilled water

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

N-ethyl-N-nitro-N-nitrosoguanidine

benzo(a)pyrene

methylmethanesulfonate

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: The bacteria were grown overnight in a shaking water bath for 16 hours at 37°C in 2.5% Nutrient Broth No. 2. After centrifugation, the bacteria were resuspended to a concentration of approximately 1 x 10E8 to 2 x 10E9 cells per millilitre in 0.16% Nutrient Broth and 0.5% Sodium chloride. The concentration of germs was controlled photometrically and determined in an experimental test with Histidine-rich potassium chloride solution on selective agar plates.

DETERMINATION OF CYTOTOXICITY
- To estimate the toxicity of the test material prototrophic bacteria (spontaneous revertants of TA 1537 = RTA) were added as an internal standard of to the selective agar plates together with TA 1537. The difference in the number of colonies on plates with and without added prototrophic bacteria at each dose level of the test material was compared to the number of colonies obtained with the negative control. The ratio of the two values was expressed as relative survival rate with the negative control at a 100 % survival rate.
Additionally, the toxicity of the test material can be evidenced in the mutagenicity assay, by a reduction in the number of revertants or by a clearing of the background lawn.

Evaluation criteria:

A material is identified as a mutagen in this test system if there is a reproducible demonstration of a dose effect relation with a 2-fold increase in the number of revertants over the controls in at least one strain . With the strain TA 100 a 1.5-fold increase is the criterion for a positive result .

Statistics:

The difference in the number of colonies on plates with and without added prototrophic bacteria at each dose level of the test material was compared to the number of colonies obtained with the negative control. The ratio of the two values was expressed as relative survival rate with the negative control at a 100 % survival rate.
Additionally, the toxicity of the test material can be evidenced in the mutagenicity assay, by a reduction in the number of revertants or by a clearing of the background lawn.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

(No relevant increase of the revertant colony numbers was obtained in Salmonella typhimurium strain used at all dose levels tested in comparison with the corresponding controls. The presence of microsomal activation did not influence these findings.)

Cytotoxicity / choice of top concentrations:

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

Remarks:

(In the toxicity experiment with 3-Sulfopropyl-methacrylsauereester (SPM), neither quantitative nor qualitative evidence of a toxic effect was observed.)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

(No relevant increase of the revertant colony numbers was obtained in Salmonella typhimurium strain used at all dose levels tested in comparison with the corresponding controls. The presence of microsomal activation did not influence these findings.)

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

(In the toxicity experiment with 3-Sulfopropyl-methacrylsauereester (SPM), neither quantitative nor qualitative evidence of a toxic effect was observed.)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

In the current study, the following results have been compiled:

GERM CONCENTRATIONS :
The concentrations of Salmonella typhimurium used in this assay were as follows:
TA 98 = 9.8 x 10 exp. 8 cells/ml
TA 100 = 5.7 x 10 exp. 8 cells/ml
TA 1535 = 1.5 x 10 exp. 9 cells/ml
TA 1537 = 9.9 x 10 exp. 8 cells/ml
TA 1530 = 1.3 x 10 exp. 9 cells/ml
in the experiment without metabolic activation,

TA 98 = 1.5 x 10 exp. 9 cells/ml
TA 100 = 7.4 x 10 exp. 8 cells/ml
TA 1535 = 1.3 x 10 exp. 9 cells/ml
TA 1537 = 1.5 x 10 exp. 9 cells/ml
TA 1538 = 1.3 x 10 exp. 9 cells/ml
in the experiment with metabolic activation.

Control plates with the used solvent (negative control) showed numbers of spontaneous revertant colonies per plate which were within the normal range of those cited in the literature .
Control plates with reference mutagens (positive controls) showed a distinct increase of the revertant colonies with the tester strains. This confirmes the reversion properties of each strain. The positive results of the mutagens 2-Amino-anthracene and Benzo(a)pyrene indicate that the metabolizing system was functioning.
The aseptic control showed no contamination for the S-9 mix.

Remarks on result:

other: all strains/cell types tested

Applicant's summary and conclusion

Conclusions:

Interpretation of results:
negative with and without metabolic activation

The study was performed according to the OECD Guideline 471 with deviations (strain TA 1538 instead of TA 102) according to the principles of the good laboratory practice and therefore considered to be of high quality (reliability Klimisch 2). The vehicle and the positive control substances fulfilled validity criteria of the test system. The test material did not induce significant increases in the frequency of revertant colonies in any of the bacterial strains. The test material was considered to be non-mutagenic under the conditions of the test.

Executive summary:

The test substance 3-Sulfopropyl-methacrylsauereester (SPM) was investigated according to OECD TG471 for its potential to cause gene mutations in Salmonella typhimurium strains (TA98, TA100, TA1537, TA1535 and TA1538). Under the conditions of this experiment 3-Sulfopropyl-methacrylsauereester (SPM) was found to cause no toxic effects. Up to the highest investigated dose, no relevant increase of the revertant colony numbers was obtained in any Salmonella typhimurium strain in comparison with the corresponding controls. The presence of microsomal activation did not influence these findings. In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test material caused neither base-pair substitutions, nor frameshift mutations. Therefore the test results with 3-Sulfopropyl-methacrylsauereester (SPM) revealed no indication of gene mutagenic activity.