FINAL REPORT 

Study Title

ACUTE TOXICITY STUDY IN DAPHNIA MAGNA WITH

1-PROPANESULFONIC ACID, 2-HYDROXY-3-(2-PROPEN-1-

YLOXY)-, SODIUM SALT (1:1)

(STATIC)

Author

L.M. Bouwman, MSc.

Test Facility

WIL Research Europe B.V.

Hambakenwetering 7

5231 DD ‘s-Hertogenbosch

The Netherlands

 

Laboratory Project Identification

Project 500676

Substance 204240/A

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1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500676

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1. CONTENTS

1. CONTENTS ...................................................................................................................................... 2

2. STATEMENT OF GLP COMPLIANCE............................................................................................. 3

3. QUALITY ASSURANCE STATEMENT ............................................................................................ 4

4. SUMMARY........................................................................................................................................ 5

5. INTRODUCTION .............................................................................................................................. 6

5.1. Preface...................................................................................................................................... 6

5.2. Aim of the study ........................................................................................................................ 6

5.3. Guidelines ................................................................................................................................. 6

5.4. Storage and retention of records and materials........................................................................ 6

5.5. Definitions.................................................................................................................................. 6

6. MATERIALS AND METHODS .......................................................................................................... 7

6.1. Test Substance ......................................................................................................................... 7

6.1.1. Test substance information............................................................................................. 7

6.1.2. Study specific test substance information ...................................................................... 7

6.1.3. Reference substance...................................................................................................... 7

6.2. Test system............................................................................................................................... 7

6.3. Breeding.................................................................................................................................... 8

6.4. Preparation of test solutions...................................................................................................... 9

6.5. Range-finding test ..................................................................................................................... 9

6.6. Limit test.................................................................................................................................... 9

6.6.1. Test concentrations......................................................................................................... 9

6.6.2. Test procedure and conditions ....................................................................................... 9

6.6.3. Sampling for analysis of test concentrations ................................................................10

6.6.4. Measurements and recordings .....................................................................................10

6.7. Electronic data capture ...........................................................................................................10

6.8. Interpretation ...........................................................................................................................10

6.8.1. Data handling................................................................................................................10

6.8.2. Acceptability of the test.................................................................................................10

6.9. List of deviations .....................................................................................................................10

6.9.1. List of protocol deviations .............................................................................................10

6.9.2. List of standard operating procedures deviations.........................................................11

7. RESULTS .......................................................................................................................................11

7.1. Range-finding test ...................................................................................................................11

7.2. Limit test..................................................................................................................................11

7.2.1. Measured concentrations .............................................................................................11

7.2.2. Immobility......................................................................................................................11

7.2.3. Determination of effect concentrations .........................................................................12

7.2.4. Experimental conditions................................................................................................12

8. CONCLUSION................................................................................................................................13

TABLES

Table 1 Incidence of immobility in the range-finding test...........................................................11

Table 2 Acute immobilisation of daphnids after 24 and 48 hours in the limit test......................12

Table 3 Effect parameters..........................................................................................................12

Table 4 pH and oxygen concentrations during the limit test......................................................12

APPENDICES 

APPENDIX 1 REFERENCE TEST......................................................................................................14

APPENDIX 2 CERTIFICATE OF ANALYSIS......................................................................................15

APPENDIX 3 ANALYTICAL REPORT................................................................................................16

Final Report


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4. SUMMARY

Acute Toxicity Study in Daphnia magna with 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, 

sodium salt (1:1). 

The study procedure described in this report was based on the OPPTS 850.1010, 1996. In addition, 

the procedures were designed to meet the test methods of the OECD guideline No. 202, 2004. 

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.

A limit test was performed based on the results of a preceding range-finding test. 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 final test 

concentration was clear and colourless.

Twenty daphnids per concentration (four replicates, five daphnids per replicate) were exposed to a 

nominal concentration of 1000 mg/l and a control. The total test period was 48 hours and samples for 

analytical confirmation of actual exposure concentrations were taken at the start and the end of the 

test.

Analysis of the sample taken at the start of the test showed that the actual test concentration was in 

agreement with nominal (100%). 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.

The study met the acceptability criteria prescribed by the protocol and was considered valid.

1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) did not induce acute 

immobilisation of Daphnia magna at an analytically confirmed nominal concentration of 1000 mg/l.

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

Hence, the 48h-EC50 was >1000 mg test substance/l, corresponding to >386 mg a.i./l. 

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5. INTRODUCTION

5.1. Preface

Sponsor Society of Chemical Manufacturers and Affiliates

1850 M Street, NW Suite 700

WASHINGTON, DC 20036

USA

Study Monitor Dr. C. T. Helmes

Test Facility WIL Research Europe B.V.

Hambakenwetering 7

5231 DD 's-Hertogenbosch

The Netherlands

Study Director L.M. Bouwman, MSc.

Principal Scientist M.J.C. Brekelmans, MSc.

Study Plan Start : 09 April 2013

Completion : 15 May 2013

5.2. Aim of the study

The purpose of the study was to evaluate the test substance for its ability to generate acute toxic 

effects on the mobility of Daphnia magna during an exposure period of 48 hours and, if possible, to 

determine the EC50 at 24 and 48 hours of exposure.

5.3. Guidelines

The study procedure described in this report was based on the EPA Ecological Effects Test 

Guidelines, OPPTS 850.1010, Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids. ‘Public 

Draft’, EPA 712-C-96-114, April 1996.

In addition, the procedure was designed to meet the test methods prescribed by the following 

guideline:

• OECD guidelines for Testing of Chemicals, guideline No. 202: "Daphnia sp., Acute Immobilisation

Test", Adopted April 13, 2004. 

5.4. Storage and retention of records and materials

Records and materials pertaining to the study including protocol, raw data, specimens (except 

specimens requiring refrigeration or freezing) and the final report are retained in the WIL Research 

Europe archives for a period of at least 2 years after finalization of the report. After this period, the 

sponsor will be contacted to determine how the records and materials should be handled. WIL 

Research Europe will retain information concerning decisions made.

Those specimens requiring refrigeration or freezing will be retained by WIL Research Europe for as 

long as the quality of the specimens permits evaluation but no longer than three months after 

finalization of the report.

WIL Research Europe will retain a test substance sample until the expiry date, but no longer than 10 

years after finalization of the report. After this period the sample will be destroyed. 

5.5. Definitions

Immobile are those animals not able to swim within 15 seconds after gentle agitation of the test 

vessel.

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The EC50 is the concentration of test substance estimated to immobilise 50% of the daphnids after a 

defined period of exposure.

6. MATERIALS AND METHODS 

6.1. Test Substance

6.1.1. Test substance information

Identification 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-

yloxy)-, sodium salt (1:1)

Synonyms COPS_1 or CS-AHPS

Structure

O SO3

-

OH

Na+

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 Percent

52556-42-0 38.64

123-34-2 2.26

7732-18-5 59.1

Test substance storage At room temperature in the dark

Stability under storage conditions Stable

Expiry date 13 August 2013

See APPENDIX 2: Certificate of analysis. 

6.1.2. Study specific test substance information

Purity/composition correction factor 

required

No 

Hygroscopic No

Volatile No

Specific Gravity / Density 1.17

Stability in water 1 year

6.1.3. Reference substance

This report includes the results of the most recent reference test with potassium dichromate 

(APPENDIX 1).

6.2. Test system 

Species Daphnia magna (Crustacea, Cladocera) (Straus, 1820), at 

least third generation, obtained by acyclical parthenogenesis 

under specified breeding conditions.

Source In-house laboratory culture with a known history.

Reason for selection This system has been selected as an internationally 

accepted invertebrate species.

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Validity of batch Daphnids originated from a healthy stock, 2nd to 5th brood, 

showing no signs of stress such as mortality >20%, 

presence of males, ephippia or discoloured animals and 

there was no delay in the production of the first brood.

Characteristics For the test selection of young daphnids with an age of < 24 

hours, from parental daphnids of more than two weeks old.

6.3. Breeding

Start of each batch With newborn daphnids, i.e. less than 3 days old, by placing 

about 250 of them into 5 litres of medium in an all-glass 

culture vessel.

Maximum age of the cultures 4 weeks

Renewal of the cultures After 7 days of cultivation half of the medium twice a week.

Temperature of medium 18-22°C

Feeding Daily, a suspension of fresh water algae.

Medium M7, as prescribed by Dr. Elendt-Schneider

(Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An 

ultrastructural approach to antennal damage in Daphnia 

magna Straus. Protoplasma 154, 25-33).

Composition of medium M7: 

Adjusted ISO medium: the following chemicals (analytical grade) are dissolved in tap water purified by Reverse Osmosis (ROwater, GEON Waterbehandeling, Berkel-Enschot, The Netherlands):

Macro salts: CaCl2.2H2O 211.5 mg/l

 MgSO4.7H2O 88.8 mg/l 

 NaHCO3 46.7 mg/l 

 KCl 4.2 mg/l

Medium M7: trace elements, macro nutrients and vitamins are added to freshly prepared ISO medium to reach the following 

concentrations: 

Trace elements: B 0.125 mg/l 

 Fe 0.05 mg/l

 Mn 0.025 mg/l

 Li, Rb and Sr 0.0125 mg/l

 Mo 0.0063 mg/l

Br 0.0025 mg/l

 Cu 0.0016 mg/l

 Zn 0.0063 mg/l

 Co and I 0.0025 mg/l 

Se 0.0010 mg/l

 V 0.0003 mg/l

 Na2EDTA.2H2O 2.5 mg/l 

 Macro nutrients: Na2SiO3. 9H2O 10.0 mg/l 

NaNO3 0.27 mg/l

 KH2PO4 0.14 mg/l

 K2HPO4 0.18 mg/l

 Vitamins: Thiamine 75.0 μg/l 

 B12 1.0 μg/l

 Biotin 0.75 μg/l

The hardness: 180 mg/l expressed as CaCO3 and the pH: 7.7 ± 0.3.

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6.4. Preparation of test solutions

The standard test procedures required generation of test solutions, which should contain completely 

dissolved test substance concentrations or stable and homogeneous mixtures or dispersions. The 

testing of concentrations that disturb the test system should be prevented (e.g. film of the test 

substance on the water surface). No correction was made for the purity/composition of the test 

substance. 

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 test were prepared by subsequent 

dilutions of the 1000 mg/l concentration in test medium. The final test solutions were all clear and 

colourless.

6.5. Range-finding test

A range-finding test was performed to provide information about the range of concentrations to be 

used in the final test. Test procedure and conditions were similar to those applied in the limit test with 

the following exceptions: 

• Ten daphnids per concentration (in duplicate, 5 per vessel) were exposed to nominal 

concentrations of 0.10, 1.0, 10, 100 and 1000 mg/l, containing 0.039, 0.39, 3.9, 39 and 

386 mg a.i. per litre, respectively; 

• Dissolved oxygen concentrations and pH were only measured in the control and the highest 

test concentration; 

• No sampling for determination of actual test concentrations was performed.

6.6. Limit test

6.6.1. Test concentrations

1-Propanesulfonic acid, 1000 mg/l, containing 386 mg a.i. per litre

2-hydroxy-3-(2-propen-1-yloxy)-, 

sodium salt (1:1) 

Control Test medium without test substance or other additives 

6.6.2. Test procedure and conditions

Test duration 48 hours

Test type Static

Test vessels 100 ml, all-glass 

Medium Adjusted ISO medium 

Number of daphnids 20 per concentration

Loading 5 per vessel containing 80 ml of test solution

Light 16 hours photoperiod daily

Feeding No feeding 

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Aeration No aeration of the test solutions.

Introduction of daphnids Within 20 minutes after preparation of the test solutions.

6.6.3. Sampling for analysis of test concentrations

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

the schedule below. The method of analysis is described in the appended Analytical Report 

(APPENDIX 3). 

Frequency at t=0 h and t=48 h 

Volume 1.5 ml from the approximate centre of the test vessels 

Storage Samples were stored in a freezer until analysis.

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

Additionally, reserve samples of 1.5 ml were taken for possible analysis. If not 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.

6.6.4. Measurements and recordings

Immobility (including mortality) At 24 hours and at 48 hours.

pH and dissolved oxygen At the beginning and at the end of the test, for both the 

control and the limit concentration.

Temperature of medium Continuously in a temperature control vessel, beginning at 

the start of the test.

6.7. Electronic data capture

Observations/measurements in the study were recorded electronically using the following 

programme(s):

- REES Centron Environmental Monitoring system version SQL 2.0 (REES Scientific, Trenton, NJ, 

USA): Temperature.

6.8. Interpretation

6.8.1. Data handling

No EC50 could be calculated because the test substance proved to be non-toxic (EC50 > maximum 

concentration tested).

6.8.2. Acceptability of the test

1. In the control, no daphnids became immobilised.

2. The oxygen concentration at the end of the test was ≥3 mg/l in control and test vessels.

6.9. List of deviations

6.9.1. List of protocol deviations

There were no deviations from the protocol. 

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6.9.2. List of standard operating procedures deviations

Any deviations from standard operating procedures were evaluated and filed in the study file. There 

were no deviations from standard operating procedures that affected the integrity of the study. 

7. RESULTS

7.1. Range-finding test

Table 1 shows the responses recorded during the range-finding test. No immobility was observed at 

any of the test concentrations during the test period. Therefore, the expected EC50 was above a 

nominal concentration of 1000 mg/l, containing 386 mg a.i. per litre. 

All test conditions were maintained within the limits prescribed by the protocol.

Table 1 Incidence of immobility in the range-finding test

Nominal conc. 

Test substance1

(mg/l)

Vessel 

number

Number

Daphnia

exposed

Response at 24 h Response at 48 h

number

Total

% number

Total

%

control A 

B

5 

5

0 

0 0 0 0 0 

0.10 A 

B

5 

5

0 

0 0 0 0 0 

1.0 A 

B

5 

5

0 

0 0 0 0 0 

10 A 

B

5 

5

0 

0 0 0 0 0 

100 A 

B

5 

5

0 

0 0 0 0 0 

1000 A 

B

5 

5

0 

0 0 0 0 0 

1 Test substance is 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1)

7.2. Limit test

7.2.1. Measured concentrations

The results of analysis of the samples taken during the limit test are described in Table 2 of the

appended Analytical Report.

Analysis of the sample taken at the start of the test showed that the actual test concentration was in 

agreement with nominal (100%). 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.

7.2.2. Immobility

Table 2 shows the responses recorded during the limit test. No immobility was observed in the limit 

concentration during the test period. The responses recorded in this test allowed for reliable 

determination of an EC50. 

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Table 2 Acute immobilisation of daphnids after 24 and 48 hours in the limit test

Concentration

Test substance1

(mg/l)

Vessel

number

Number

Daphnia

exposed

Response at 24 h Response at 48 h

number

Total

% number

Total

%

control

A 

B 

C 

D

5 

5 

5 

5

0 

0 

0 

0

0 0 

0 

0 

0

0 

1000

A 

B 

C 

D

5 

5 

5 

5

0 

0 

0 

0

0 0 

0 

0 

0

0 

1 Test substance is 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1)

7.2.3. Determination of effect concentrations

Table 3 shows the effect parameters based on the analytically confirmed nominal concentration of the 

test substance and on the corresponding concentration of the active ingredient. 

Table 3 Effect parameters

Parameter Concentration

Test substance1 

(mg/l)

Concentration

Active ingredient2 

(mg/l)

24& 48h-EC50 >1000 >386 1 Test substance is 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) 2 Active ingredient is Sodium-1-allyloxy-2-hydroxypropane sulphonate

7.2.4. Experimental conditions

The results of measurement of pH and oxygen concentrations (mg/l) are presented in Table 4. These 

test conditions remained within the limits prescribed by the protocol (pH: 6.0-8.5, not varying by more 

than 1.5 unit; oxygen: ≥3 mg/l at the end of the test).

The temperature of the test medium ranged between 20.3 and 20.5°C at the start of the test. The 

temperature continuously measured in a temperature control vessel varied between 19.4 and 20.3°C 

during the test, and complied with the requirements as laid down in the protocol (18-22°C, constant 

within 2°C).

Table 4 pH and oxygen concentrations during the limit test

Concentration 

Test substance1

 

(mg/l)

Start (t=0 h) End (t=48 h)

pH O2 pH O2

control 7.8 9.2 7.9 8.8

1000 7.6 9.2 7.8 8.8 1 Test substance is 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1)

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8. CONCLUSION

Under the conditions of the present study 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, 

sodium salt (1:1) did not induce acute immobilisation of Daphnia magna at an analytically confirmed 

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

litre.

Hence, the 48h-EC50 was >1000 mg test substance/l, corresponding to >386 mg a.i./l. 

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APPENDIX 1 REFERENCE TEST

Start: 04 March 2013

End : 06 March 2013

48-hour Acute Toxicity Study in Daphnia magna with potassium dichromate (K2Cr2O7) (WIL Research 

Europe Project 502370).

The study procedures described in this report were based on the OECD guideline No. 202: "Daphnia 

sp., Acute Immobilisation Test", Adopted April 13, 2004 and the ISO International Standard 6341.

The reference test was carried out to check the sensitivity of the test system as used by WIL Research 

Europe. Daphnia were exposed for a maximum of 48 hours to K2Cr2O7 concentrations of 0.10, 0.18, 

0.32, 0.56, 1.0 and 1.8 mg/l and to a control. Twenty daphnia were exposed per concentration.

The reference substance, potassium dichromate (K2Cr2O7, art. 1.04864, batch no. K34869764 607) 

was obtained from Merck, Darmstadt, Germany.

Acute immobilization of daphnia after 24 and 48 hours in the reference test with potassium 

dichromate:

K2Cr2O7

Nominal concentration 

(mg/l)

Number

Exposed

% immobile Expected response (%)

After 48 hours1

24h 48h Minimal Maximal

control 20 0 0 0 102

0.10 20 0 5 0 10

0.18 20 0 0 0 10

0.32 20 0 0 0 30

0.56 20 15 55 0 100

1.0 20 70 100 40 100

1.8 20 100 100 100 100

1: Based on historical data of the previous years (n>60).

2: A maximum response of 10% does not invalidate the results of the test.

 

The actual responses in this reference test with K2Cr2O7 are within the ranges of the expected 

responses at the different concentrations, i.e. the 48h-EC50 was between 0.3 and 1.0 mg/l. Hence, the 

sensitivity of this batch of D. magna was in agreement with the historical data collected at WIL 

Research Europe.

The 24h-EC50 was 0.78 mg/l with a 95% confidence interval between 0.69 and 0.93 mg/l.

The 48h-EC50 was 0.53 mg/l with a 95% confidence interval between 0.49 and 0.62 mg/l.

The raw data from this study are kept in the WIL Research Europe archives. The test described above 

was performed under GLP with a QA-check.

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APPENDIX 2 CERTIFICATE OF ANALYSIS

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APPENDIX 3 ANALYTICAL REPORT

DETERMINATION OF THE 

CONCENTRATIONS

Author

M.J.C. Brekelmans, MSc.

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1. CONTENTS

1. CONTENTS ................................................................................................................................... 17

2. REPORT APPROVAL.................................................................................................................... 18

3. INTRODUCTION ........................................................................................................................... 19

3.1. Preface................................................................................................................................... 19

3.2. Aim of the study ..................................................................................................................... 19

4. MATERIALS AND METHODS ....................................................................................................... 19

4.1. Reagents ................................................................................................................................ 19

4.2. Samples ................................................................................................................................. 19

4.3. Analytical method................................................................................................................... 20

4.3.1. Analytical conditions .................................................................................................... 20

4.3.2. Preparation of solutions ............................................................................................... 20

4.3.3. Sample injections......................................................................................................... 20

4.4. Electronic data capture .......................................................................................................... 20

4.5. Formulas ................................................................................................................................ 21

5. RESULTS ...................................................................................................................................... 22

5.1. Calibration curves .................................................................................................................. 22

5.2. Samples ................................................................................................................................. 22

5.2.1. Procedural recovery samples ...................................................................................... 22

5.2.2. Test samples................................................................................................................ 22

TABLES

Table 1 Procedural recovery samples ........................................................................................... 23

Table 2 Concentrations of the test substance in test medium....................................................... 23

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3. INTRODUCTION

3.1. Preface

Study plan analytical phase Start : 21 May 2013

Completion : 22 May 2013

3.2. Aim of the study

The purpose of the analytical phase was to determine the actual concentrations in samples taken from 

the test solutions used during the ecotoxicity test. 

4. MATERIALS AND METHODS 

4.1. Reagents

Water Tap water purified by a Milli-Q water purification 

system (Millipore, Bedford, MA, USA)

Di-Sodium hydrogenphosphate dihydrate Merck, Darmstadt, Germany

(Na2HPO4 · 2H2O)

Sodium dihydrogenphosphate dihydrate Merck

(NaH2PO4 · 2H2O)

Sodium nitrate Merck

(NaNO3) 

ISO-medium see main report

All reagents were of analytical grade, unless specified otherwise.

4.2. Samples

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.

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4.3. Analytical method

4.3.1. Analytical conditions

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

500674. 

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

Detector Refractive Index Detector 2414 (Waters)

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

4.3.2. Preparation of solutions

Stock and spiking solutions 

Stock solutions of the test substance were prepared in water at concentrations of 4935 and 5945 mg/l. 

Spiking solutions were made up from a stock solution and/or dilutions of this solution. The solvent of 

the spiking solutions was water.

Calibration solutions

Calibration solutions in the concentration range of 80 – 1200 mg/l were prepared from two stock 

solutions. The end solution of the calibration solutions was mobile phase. 

Procedural recovery samples

1.6 ml ISO-medium was spiked with the test substance at a target concentration of 100 or 1000 mg/l. 

The accuracy samples were analysed without further treatment.

4.3.3. Sample injections

Calibration solutions were injected in duplicate. Test samples and procedural recovery samples were 

analysed by single injection. 

4.4. Electronic data capture

System control, data acquisition and data processing were performed using the following programme:

- Empower version 7.00 (Waters, Milford, MA, USA).

Temperature, relative humidity and/or atmospheric pressure during sample storage and/or 

performance of the studies was monitored continuously using the following programme:

- REES Centron Environmental Monitoring system version SQL 2.0 (REES Scientific, Trenton, NJ, 

USA). 

Final Report
1-propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500676

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4.5. Formulas

Response (R) Peak area test substance [units]

Calibration curve N += b C a R

where:

CN = nominal concentration [mg/l]

a = slope [units × l/mg]

b = intercept [units]

Analysed concentration (CA) [mg/l] d

a

(R b) CA × − =

where:

d = dilution factor

Recovery [%] 100 C

C

N

A ×

Relative to nominal concentration [%] 100 C

C

N

A ×

Relative to initial concentration [%] 100 C (t hours) 0

 C (t hours)x 

A

A × =

=

Final Report
1-propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500676

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5. RESULTS

5.1. Calibration curves

A calibration curve was constructed using five concentrations. For each concentration, two responses 

were used. Linear regression analysis was performed using the least squares method with a 

1/concentration2 weighting factor. The coefficient of correlation (r) was > 0.99. 

5.2. Samples

5.2.1. Procedural recovery samples

The results for the procedural recovery samples are given in Table 1.

The mean recoveries of the procedural recovery samples fell within the criterion of 70-110%. It 

demonstrated that the analytical method was adequate for the determination of the test substance in 

the test samples. 

5.2.2. Test samples

The results for the test samples are given in Table 2 . 

Final Report
1-propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500676

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TABLES

Table 1 Procedural recovery samples

Date of 

preparation 

Date of 

analysis

Target 

concentration

[mg/l]

Nominal 

concentration

[mg/l]

Analysed 

concentration

[mg/l]

Recovery

[%]

Mean 

recovery

[%]

21-May-2013 21-May-2013 100 103 106 103 103

103 107 104

21-May-2013 21-May-2013 1000 1032 1048 102 102

1032 1061 103

Table 2 Concentrations of the test substance in test medium 

Time of 

sampling

[hours]

Date of 

sampling

Date of 

analysis 1

Concentration Relative to 

nominal

[%]

Relative to 

initial 

[%]

Nominal

[mg/l]

Analysed

[mg/l]

0 13-May-2013 21-May-2013 0 n.d. n.a.

1000 1000 100

48 15-May-2013 21-May-2013 0 n.d. n.a.

1000 1022 102 102

1 Samples were stored in the freezer (≤ -15°C) until the day of analysis.

n.d. Not detected.

n.a. Not applicable.

 

Final Report