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 / Nethe rlands 

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# Endpoint study record: Toxicity to aquatic algae an d

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 def ined 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., Hambakenwetering7, 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 Inhib ition 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 ana lysis. At the end of the exposure period, the replicates w ith algae were pooled at each concentration before sampling. Compliance with the Quality criteria regarding main tenance of actual concentrations was demonstrated b y running a test vessel at the highest substance concentration but without algae and sampl es for analysis were taken at the start, after 24 h ours 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 n ot already used, these samples were stored in a freezer for a maximum of three months after de livery 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 c onditions 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 m ethod validated for the test substance in project 5 00674. Analytical method Instrument: Alliance Separation Module 2695 (Waters , Milford, MA, USA) Detector: Refractive Index Detector 2414 (Waters) C olumn Column Stationary phase: Synergi Hydro-RP 80A Dimensions: 250 mm × 4.6 mm i.d., dp =4 μ m Brand: Phenomenex, Torrance, CA, USA Column temperature: 40°C ± 1°C Injection volume: 50 μ l 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 c lear colourless liquid and completely soluble in test medium at the concentration tested. Preparation of test solutions started with a nomina l 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 t o accelerate the dissolving of the test substance in the test medium. The lower test concen trations for the range-finding tests were prepared by subsequent dilutions of the 1000 mg/l concentration in test medium. The final test soluti ons were all clear and colourless.

Pseudokirchnerella subcapitata Species: Pseudokirchneriella subcapitata, strain: N IVA 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 te st, cells from the algal stock culture were inocula ted 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 d ensity 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 rev ealed a normal and healthy appearance of the expose d 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 te st showed that the actual test concentration was in agreement with nominal (101%). This concentration remained stable during the test perio d (102% of initial at the end of the test). Given t hese 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 o f test solution Medium: M2 Cell density: An initial cell density of 10000 cell s/ml. Introduction of algal cells: Immediately after prep aration of the test solutions. Illumination: Continuously using TLD-lamps of the t ype ‘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 rando m in the incubator and as such were daily repositio ned. 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 conce ntration; 1 extra replicate of both the control and the test concentration for sampling purposes; 2 replicates of the test concentration without alga e. Medium: M2 (OECD 201 guideline) Cell density: An initial cell density of 1 x 104 ce lls/ml. Introduction of algal cells: Immediately after prep aration of the test solutions. Illumination: Continuously using TLD-lamps of the t ype ‘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 rand om in the incubator and as such were daily repositi oned. 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. (analyticall y 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. (analyticall y 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 r anging from 0.62 to 1.5 mg/l. The historical ranges for growth rate reduction lie bet ween 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%). N o 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 analy tically confirmed nominal concentration of 1000 mg/l. This concentration contained 386 mg of t he active ingredient per litre. The IC50 for growth rate reduction (96h-IRC50), yie ld inhibition (96h-IYC50) and cell growth inhibitio n (96h-IBC50) was >1000 mg test substance/l, corresponding to >386 mg a.i./l. The 96h-NOEC for growth rate reduction, yield inhib ition and cell growth inhibition was 1000 mg/l. 

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