Printing Date 2013-09-02 13:19:38 UTC 

Restriction of specific regulatory purposes 

 

Confidentiality 

 

Owner (TS 204240) 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) / WIL Research Europe B.V. / 'sHertogenbosch / Netherlands 

Legal entity 

owner 

WIL Research Europe B.V. / 's-Hertogenbosch / Netherlands 

Endpoint study record: Toxicity to aquatic algae and cyanobacteria. WIL Research 50... Page 1 of 5

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Endpoint study record: Toxicity to aquatic algae and 

cyanobacteria. WIL Research 500678 

Administrative Data 

Data source 

Reference 

Materials and methods 

Test guideline 

GLP compliance 

Test materials 

Identity of test material same as for substance defined in section 1 (if not read-across) 

Test material identity 

Test material form 

Details on test material 

UUID 

 

IUC5-67803ad7-b0b9-448b-ab71-0f1826137e47 

Dossier UUID 

 

0 

Author 

 

BOL / WIL Research Europe B.V. / 's-Hertogenbosch / Netherlands 

Date 

 

2013-09-02 13:16:45 UTC 

Remarks 

 

 

Purpose flag key study; robust study summary 

Study result type experimental result Study period 08 April 2013 - 17 May 2013 

Rationale for reliability incl. 

deficiencies 

This study has been performed according to EPA and OECD guidelines and according to GLP 

principles. 

Reference 

type 

Author Year Title Bibliographic 

source 

Testing laboratory Report 

no. 

Owner 

company 

Company 

study no. 

Report 

date 

study 

report 

L.M. 

Bouwman 

2013 FRESH WATER 

ALGAL GROWTH 

INHIBITION TEST 

WITH 1-

PROPANESULFONIC 

ACID, 2-HYDROXY-3-

(2-PROPEN-1-

YLOXY)-, SODIUM 

SALT (1:1) 

 WIL Research 

Europe B.V., 

Hambakenwetering 

7, 5231 DD, 'sHertogenbosch, 

The Netherlands 

500678 Society of 

Chemical 

Manufacturers 

and Affiliates, 

1850 M Street, 

NW Suite 700, 

WASHINGTON, 

DC 20036, USA 

 

2013-

07-16 

Qualifier Guideline Deviations 

according to other guideline: OCSPP 850.4500, Algal Toxicity, EPA 712-C-006 no 

according to OECD Guideline 201 (Alga, Growth Inhibition Test) (2006) no 

yes 

yes 

Identifier Identity 

CAS number 52556-42-0; 123-34-2 and 7732-18-5 

other: Clear colourless liquid 

Molecular formula: C6H11NaO5S 

Molecular weight: 218.22 

Description: Clear colourless liquid 

Batch: 12E1101 

Purity/Composition: 

Sodium-1-allyloxy-2-hydroxypropane sulphonate 

3-allyl-1,2-propanediol 

Water 

CAS Number: 

52556-42-0 

123-34-2 

7732-18-5 

Percent: 

38.64 

2.26 

59.1 

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Analytical monitoring 

Details on sampling 

Details on analytical methods 

Vehicle 

Details on test solutions 

Test organisms 

Test organisms (species) 

Details on test organisms 

Test substance storage: At room temperature in the dark

Stability under storage conditions: Stable 

Expiry date: 13 August 2013 

Purity/composition correction factor required No 

Hygroscopic: No 

Volatile: No 

Specific Gravity / Density: 1.17 

Stability in water: 1 year 

yes 

Samples for possible analysis were taken from both the control and the test concentration according to the schedule below. 

Frequency: at t=0 h, t=24 h and t=96 h 

Volume: 1.5 ml 

Storage: Samples were stored in a freezer until analysis. 

At the end of the exposure period, the replicates with algae were pooled at each concentration before sampling. 

Compliance with the Quality criteria regarding maintenance of actual concentrations was demonstrated by running a test vessel at the highest 

substance concentration but without algae and samples for analysis were taken at the start, after 24 hours of exposure and at the end of the 

test period. 

Additionally, reserve samples of 1.5 ml were taken from all test solutions for possible analysis. If not already used, these samples were stored 

in a freezer for a maximum of three months after delivery of the draft report, pending on the decision of the sponsor for additional analysis. 

The samples were stored in the freezer (≤ -15°C). Storage stability of samples under these conditions was demonstrated in project 500674.

On the day of analysis, the samples were defrosted at room temperature and analysed. 

Quantitative analysis was based on the analytical method validated for the test substance in project 500674. 

Analytical method 

Instrument: Alliance Separation Module 2695 (Waters, Milford, MA, USA) 

Detector: Refractive Index Detector 2414 (Waters) Column 

Column 

Stationary phase: Synergi Hydro-RP 80A 

Dimensions: 250 mm × 4.6 mm i.d., dp =4 μm 

Brand: Phenomenex, Torrance, CA, USA 

Column temperature: 40°C ± 1°C 

Injection volume: 50 μl 

Mobile phase water containing: 0.05M Na2HPO4, 0.05M NaH2PO4 and 0.05M NaNO3 

Flow: 1 ml/min 

RI detection: 

Temperature: 30°C 

Sensitivity: 4 

Time constant: 1 seconds 

no 

The batch of 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) tested was a clear colourless liquid and completely 

soluble in test medium at the concentration tested.

Preparation of test solutions started with a nominal concentration of 1000 mg/l, containing 386 mg of the active ingredient (a.i.) Sodium-1-

allyloxy-2-hydroxypropane sulphonate. Approximately fifteen minutes of magnetic stirring was applied to accelerate the dissolving of the test 

substance in the test medium. The lower test concentrations for the range-finding tests were prepared by subsequent dilutions of the 1000 mg/l 

concentration in test medium. The final test solutions were all clear and colourless. 

Pseudokirchnerella subcapitata 

Species: Pseudokirchneriella subcapitata, strain: NIVA CHL 1 

Source: In-house laboratory culture. 

Reason for selection: This system is an unicellular algal species sensitive to toxic substances in the aquatic ecosystem and has been selected 

as an internationally accepted species. 

Stock culture: Algae stock cultures were started by inoculating growth medium with algal cells from a pure culture on agar. The suspensions 

were continuously aerated and exposed to light in a climate room at a temperature of 21-24°C. 

Light intensity: 60 to 120 μE/m2/s when measured in the photosynthetically effective wavelength range of 400 to 700 nm. 

Pre-culture: 3 to 4 days before the start of the test, cells from the algal stock culture were inoculated in culture medium at a cell density of 1 x 

104 cells/ml. The pre-culture was maintained under the same conditions as used in the test. The cell density was measured immediately 

before use. 

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Study design 

Test type 

Water media type 

Limit test 

Total exposure duration 

Post exposure observation period 

Test conditions 

Hardness 

Test temperature 

pH 

Dissolved oxygen 

Salinity 

Nominal and measured concentrations 

Details on test conditions 

Reference substance (positive control) 

Results and discussions 

Effect concentrations 

static 

freshwater 

yes 

96 h 

Microscopic observations at the end of the test revealed a normal and healthy appearance of the exposed cells when compared to the control. 

0.24 mmol (24 mg CaCO3/L) 

23.5 -24.9°C 

8.0-9.6 

not measured 

not measured 

Nominal: 1000 mg/l and control 

Analysis of the sample taken at the start of the test showed that the actual test concentration was in agreement with nominal (101%). This 

concentration remained stable during the test period (102% of initial at the end of the test). Given these results, the effect parameters were 

based on the nominal exposure concentration. 

Test duration: 96 hours 

Test type: Static 

Test vessels: 125 ml, all-glass, containing 50 ml of test solution 

Medium: M2 

Cell density: An initial cell density of 10000 cells/ml. 

Introduction of algal cells: Immediately after preparation of the test solutions. 

Illumination: Continuously using TLD-lamps of the type ‘Cool-white’ of 30 Watt, with a light intensity within the range of 59 to 66 μE.m-2.s-1. 

Incubation Capped vessels were distributed at random in the incubator and as such were daily repositioned. During incubation the algal cells 

were kept in suspension by continuous shaking. 

Control end cell density: 

Replicates: 

6 replicates of both the control and the test concentration; 

1 extra replicate of both the control and the test concentration for sampling purposes; 

2 replicates of the test concentration without algae. 

Medium: M2 (OECD 201 guideline) 

Cell density: An initial cell density of 1 x 104 cells/ml. 

Introduction of algal cells: Immediately after preparation of the test solutions. 

Illumination: Continuously using TLD-lamps of the type ‘Cool-white’ of 30 Watt, with a light intensity within the range of 59 to 66 micro E.m-2.s1. 

Incubation: Capped vessels were distributed at random in the incubator and as such were daily repositioned. During incubation the algal cells 

were kept in suspension by continuous shaking. 

Control end cell density: 2709000 cells/ml 

yes Potassium dichromate (K2Cr2O7) 

Duration Endpoint Effect 

conc. 

Nominal/Measured Conc. based on Basis for 

effect 

Remarks (e.g. 95% CL) 

96 h NOEC 1000 

mg/L 

nominal test mat. (analytically confirmed 

nominal concentration) 

growth rate NOEC: >386 mg active 

ingredient /L 

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Details on results 

Results with reference substance (positive control) 

Reported statistics and error estimates 

Applicant's summary and conclusion 

Validity criteria fulfilled 

Conclusions 

96 h IC50 > 1000 

mg/L 

nominal test mat. (analytically confirmed 

nominal concentration) 

growth rate IC50: >386 mg active 

ingredient /L 

96 h NOEC 1000 

mg/L 

nominal test mat. (analytically confirmed 

nominal concentration) 

other: yield NOEC: >386 mg active 

ingredient /L 

96 h IC50 > 1000 nominal test mat. (analytically confirmed 

nominal concentration) 

other: yield IC50: >386 mg active 

ingredient /L 

96 h NOEC 1000 

mg/L 

nominal test mat. (analytically confirmed 

nominal concentration) 

biomass (cell 

growth) 

NOEC: >386 mg active 

ingredient /L 

96 h IC50 > 1000 nominal test mat. (analytically confirmed 

nominal concentration) 

biomass (cell 

growth) 

IC50: >386 mg active 

ingredient /L 

-Exponential growth in the control :yes

EC50: The EC50 for growth rate reduction (ERC50: 0-72h) was 0.97 mg/l with a 95% confidence interval ranging from 0.62 to 1.5 mg/l. The 

historical ranges for growth rate reduction lie between 0.82 and 2.3 mg/l. Hence, the ERC50: 0-72h for the algal culture tested corresponds 

with this range. 

Statistical analysis of the data was not needed as the effects recorded were not significant (<10%). No IC50-values could be calculated 

because the test substance proved to be non-toxic (IC50 > maximum concentration tested). 

yes 

No reduction of growth rate, inhibition of yield or inhibition of cell growth was recorded at an analytically confirmed nominal concentration of 

1000 mg/l. This concentration contained 386 mg of the active ingredient per litre. 

The IC50 for growth rate reduction (96h-IRC50), yield inhibition (96h-IYC50) and cell growth inhibition (96h-IBC50) was >1000 mg test 

substance/l, corresponding to >386 mg a.i./l. 

The 96h-NOEC for growth rate reduction, yield inhibition and cell growth inhibition was 1000 mg/l. 

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