FINAL REPORT 

Study Title

96-HOUR ACUTE TOXICITY STUDY IN FATHEAD MINNOW 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 500677

Substance 204240/A

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1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500677

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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. Holding ...................................................................................................................................... 8

6.4. Preparation of test solutions...................................................................................................... 8

6.5. Limit test.................................................................................................................................... 9

6.5.1. Test concentrations......................................................................................................... 9

6.5.2. Test procedure and conditions ....................................................................................... 9

6.5.3. Sampling for analysis of test concentrations .................................................................. 9

6.5.4. Measurements and recordings .....................................................................................10

6.6. Electronic data capture ...........................................................................................................10

6.7. Interpretation ...........................................................................................................................10

6.7.1. Data handling ................................................................................................................10

6.7.2. Acceptability of the test.................................................................................................10

6.8. List of deviations .....................................................................................................................10

6.8.1. List of protocol deviations .............................................................................................10

6.8.2. List of standard operating procedures deviations.........................................................10

7. RESULTS .......................................................................................................................................10

7.1. Measured concentrations........................................................................................................10

7.2. Mortality and other effects.......................................................................................................11

7.3. Determination of effect concentrations ...................................................................................11

7.4. Experimental conditions ..........................................................................................................11

8. CONCLUSION................................................................................................................................12

TABLES

Table 1 Incidence of mortality and total mortality ......................................................................11

Table 2 Effect parameters..........................................................................................................11

Table 3 pH-values and dissolved oxygen concentrations (mg/l) ...............................................11

Table 4 Temperatures (°C)........................................................................................................11

APPENDICES 

APPENDIX 1 REFERENCE TEST......................................................................................................13

APPENDIX 2 CERTIFICATE OF ANALYSIS......................................................................................14

APPENDIX 3 ANALYTICAL REPORT................................................................................................15

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4. SUMMARY

96-Hour Acute Toxicity Study in Fathead minnow with 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen1-yloxy)-, sodium salt (1:1). 

The study procedure described in this report was based on the OPPTS 850.1075, 1996. In addition, 

the procedures were designed to meet the test methods of the OECD guideline No. 203, 1992. 

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. 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.

A limit test was performed exposing fourteen fish (two replicates, seven fish per replicate) to a nominal 

concentration of 1000 mg/l. Simultaneously, seven fish were exposed to a control. The total test period 

was 96 hours and samples for analytical confirmation of actual exposure concentrations were taken at 

the start, after 48 hours of exposure and at the end of the test.

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

agreement with nominal (102-103%). This concentration remained stable during the test period (99-

101% 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) induced no visible effects in 

fathead minnow at an analytically confirmed nominal concentration of 1000 mg/l. This concentration 

contained 386 mg of the active ingredient per litre.

Hence, the 96h-LC50 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 : 13 May 2013

Completion : 17 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 in Pimephales promelas during an exposure period of 96 hours and, if possible, to determine 

the LC50 at all observation times.

5.3. Guidelines

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

Guidelines, OPPTS 850.1075, Fish Acute Toxicity Test, Freshwater and marine. ‘Public Draft’, 

EPA 712-C-96-118, 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. 203: "Fish Acute Toxicity Test", Adopted 

17 July, 1992.

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 limit report are retained in the WIL Research 

Europe archives for a period of at least 2 years after finalisation 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 

finalisation of the report.

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

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

5.5. Definitions

Fish were considered to be dead when no reaction was observed after touching the caudal peduncle

and visible breathing movements were absent. In addition, fish that were convulsing or showing other 

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severe forms of distress not considered transient in nature and likely to become more severe before 

the exposure is terminated, will be sacrificed for humane reasons. These fish will be treated as having 

died in the test, based on OECD guideline on humane endpoint (ENV/JM/MONO/(2000)7).

The LC50 is the concentration killing 50% of the fish 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 a reference test with pentachlorophenol (PCP) (APPENDIX 1).

6.2. Test System

Species Fathead minnow (Pimephales promelas, Teleostei, 

Cyprinidae) Rafinesque

Source In house culture (Batch FM 13-02)

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Mean length1 1.9 ± 0.1 cm 

Mean weight1 0.14 ± 0.04 g 

Reason for selection This system has been selected as an internationally 

accepted species.

Total fish used 21

6.3. Holding 

Medium Adjusted ISO medium, formulated using RO-water (tapwater purified by reverse osmosis; GEON 

Waterbehandeling, Berkel-Enschot, The Netherlands) with 

the following composition:

CaCl2.2H2O 211.5 mg/l

MgSO4.7H2O 88.8 mg/l

NaHCO3 46.7 mg/l

KCl 4.2 mg/l

Measurements Conductivity, pH, nitrate, nitrite and ammonia concentration: 

once a week. Temperature: continuous. In addition, pH and 

temperature were measured before transferring the fish to 

the test system.

Water quality parameters Were kept within the optimum limits for the respective fish 

species.

Feeding Daily with pelleted fish food (Cyprico Crumble Excellent 

(300-500 um), Coppens International bv, Helmond, The 

Netherlands) 

Validity of batch In the batch of fish used for the test, mortality during the 

seven days prior to the start of the test was less than 5%.

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. 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.

 

1 Ten fish of the batch used for the test, were weighed and measured prior to the start of the test.

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6.5. Limit test

6.5.1. 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 

6.5.2. Test procedure and conditions 

Test duration 96 hours

Test type Static

Test vessels 6.5 litres, all-glass, containing 5 litres of test solution

 

Test medium Adjusted ISO medium with a hardness of 180 mg CaCO3 per 

litre and a pH of 7.7 ± 0.3.

Number of fish 7 for the control and 14 for the test concentration

Number of replicates 1 for the control;

2 for the test concentration, containing 7 fish each

Loading 0.20 g fish/litre, i.e. 7 fish per 5 litres of test medium

Illumination 16 hours photoperiod daily

Aeration The test media were not aerated during the test.

Feeding No feeding from 24 hours prior to the test and during the 

total test period

Introduction of fish Within 40 minutes after preparation of the test media from a 

holding tank with comparable water quality parameters and 

pH and temperature differences between test and holding 

tank media of less than 0.5 unit and 0.5°C. 

Euthanasia At the end of the test the surviving fish were rapidly killed by 

exposing them to ca. 1.2% ethylene glycol monophenylether 

in water.

6.5.3. Sampling for analysis of test concentrations

Samples for possible analysis were taken from the control and both replicates of 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, t=48 h and t=96 h 

Volume 1.5 ml

Storage Samples were stored in a freezer until analysis.

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.

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6.5.4. Measurements and recordings 

Mortality and other effects At 6, 24, 48, 72 and 96 hours following the start of exposure. 

In addition, every afternoon and morning from day 1, except 

for the afternoon at day 2, to observe for any dead or 

severely distressed fish. 

Dissolved oxygen content, Daily in all vessels with surviving fish, beginning at the

pH and temperature start of the test (day 0).

6.6. Electronic data capture

Temperature during sample storage was monitored continuously using the following programme:

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

USA)

6.7. Interpretation

6.7.1. Data handling 

No LC50 could be calculated because the test substance proved to be non-toxic (LC50 > maximum 

concentration tested).

6.7.2. Acceptability of the test

1. No mortality was observed in the control at the end of the test.

2. Test conditions were maintained constant throughout the test.

3. The dissolved oxygen concentration has been at least 60% of the air saturation value throughout 

the test (>5 mg/l at 22°C).

4. The concentrations of the substance being tested have been at least 80% of the nominal 

concentration throughout the test.

6.8. List of deviations

6.8.1. List of protocol deviations

1. Fish mortality was not recorded at the end of the second day of exposure. 

Evaluation: This is a requirement for the Ethical Committee, not for the endpoints of this study.

The study integrity was not adversely affected by this deviation.

6.8.2. 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. 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 samples taken at the start of the test showed that the actual test concentration was in 

agreement with nominal (102-103%). This concentration remained stable during the test period (99-

101% of initial at the end of the test). Given these results, the effect parameters were based on the 

nominal exposure concentration.

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7.2. Mortality and other effects

Table 1 shows the mortality data recorded during the limit test. No mortality or other clinical effects 

were observed in both the control and the test concentration. The responses recorded in this test 

allowed for reliable determination of an LC50. 

Table 1 Incidence of mortality and total mortality

Concentration

Test substance1

(mg/l)

Vessel

number

Initial

number

of fish

Cumulative mortality Total

Mortality

(%) 6h 24h 48h 72h 96h

control A 7 0 0 0 0 0 0

1000 A 7 0 0 0 0 0 0 B 7 0 0 0 0 0 1 Test substance is 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1)

7.3. Determination of effect concentrations

Table 2 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 2 Effect parameters

Parameter Concentration 

Test substance1 

(mg/l)

Concentration 

Active ingredient2 

(mg/l)

24, 48, 72 & 96h-LC50 >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

7.4. Experimental conditions

The results of measurement of pH and oxygen concentrations are presented in Table 3. The 

temperatures measured during the study in the various test vessels are presented in Table 4. All test 

conditions remained within the ranges prescribed by the protocol (pH: 6.0-8.5, constant within 1 unit; 

temperature 20-24°C, constant within 2°C; oxygen > 60% of air saturation). 

Table 3 pH-values and dissolved oxygen concentrations (mg/l)

Concentration

Test substance1 

(mg/l)

Vessel

number

Day 0 Day 1 Day 2 Day 3 Day 4

pH2 O2 pH O2 pH O2 pH O2 pH O2

control A 7.9 9.2 7.8 7.7 7.6 7.2 7.6 7.1 7.8 7.1

1000 A 7.7 9.0 7.7 7.6 7.6 7.1 7.6 6.9 7.7 6.9

B 7.6 9.1 7.6 7.4 7.5 7.0 7.5 6.8 7.7 6.8 1 Test substance is 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) 2 pH of culture medium was: 7.7 

Table 4 Temperatures (°C)

Concentration

Test substance1 

(mg/l)

Vessel

Number

Day 02 Day 1 Day 2 Day 3 Day 4

control A 22.8 22.6 22.7 22.5 22.7

1000 A 22.9 22.8 22.8 22.8 22.7

B 23.0 22.9 22.8 22.8 22.8 1 Test substance is 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) 2 Temperature of culture medium was: 23.3°C. Therefore, the fish were acclimatized to 22.9°C before the start of the test.

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8. CONCLUSION

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

sodium salt (1:1) induced no visible effects in fathead minnow at an analytically confirmed nominal 

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

Hence, the 96h-LC50 was >1000 mg test substance/l, corresponding to >386 mg a.i./l.

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APPENDIX 1 REFERENCE TEST

96-hour acute toxicity study in the Fathead minnow with pentachlorophenol (PCP); WIL Research 

Europe Project 502823 (Batch FM 13-02).

The study procedure described in this report was based on the OECD guideline No. 203, 1992. In 

addition, the procedures were designed to meet the test methods of the ISO International Standard 

7346-1: Static method, 1996.

Start: 13 May 2013

End: 17 May 2013

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

Research Europe. The reference substance was pentachlorophenol (PCP, SIGMA, Art. P2604, Batch 

06324ED-405). 

Concentrations: 0.10, 0.22 and 0.46 mg/l in ISO-medium. 

Incidence of mortality observed in the reference study:

Concentration

PCP (mg/l)

Nominal

Initial

Number

Of fish

Cumulative number of dead fish recorded at 

various time points after start of exposure

Total

Mortality

(%) 13⁄4h 24h 48h 72h 96h

0.10 5 0 0 0 0 0 0

0.22 5 0 3 3 3 3 60

0.46 5 0 4 4 5 5 100

During the test the pH, oxygen concentration and the temperature of the medium were within the 

optimal ranges for fish.

Under the conditions of the present test with Fathead minnow exposed to PCP, the 96h-LC50 was 

0.20 mg/l based on nominal concentrations, with a 95% confidence interval between 0.16 and 

0.32 mg/l. This effect was already reached within 72 hours of exposure.

No historical data is available on the range of the 96h-LC50 for Fathead minnow. However, this test 

shows that toxicity is similar for Fathead minnow and carp (Teleostei Cyprinidae). For Carp the 

96h-LC50 is generally between 0.10 and 0.46 mg/l based on historical data of reference tests 

performed approximately every 3 months from April 1988 until the end of 2000, and annually since 

then.

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

described above was performed under GLP-conditions 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 ................................................................................................................................... 16

2. REPORT APPROVAL.................................................................................................................... 17

3. INTRODUCTION ........................................................................................................................... 18

3.1. Preface................................................................................................................................... 18

3.2. Aim of the study ..................................................................................................................... 18

4. MATERIALS AND METHODS ....................................................................................................... 18

4.1. Reagents ................................................................................................................................ 18

4.2. Samples ................................................................................................................................. 18

4.3. Analytical method................................................................................................................... 19

4.3.1. Analytical conditions .................................................................................................... 19

4.3.2. Preparation of solutions ............................................................................................... 19

4.3.3. Sample injections......................................................................................................... 19

4.4. Electronic data capture .......................................................................................................... 19

4.5. Formulas ................................................................................................................................ 20

5. RESULTS ...................................................................................................................................... 21

5.1. Calibration curves .................................................................................................................. 21

5.2. Samples ................................................................................................................................. 21

5.2.1. Procedural recovery samples ...................................................................................... 21

5.2.2. Test samples................................................................................................................ 21

TABLES

Table 1 Procedural recovery samples ........................................................................................... 22

Table 2 Concentrations of the test substance in test medium....................................................... 22

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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). 

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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 × =

=

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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 . 

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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 1034 2 103

1000 1024 3 102

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

1000 1011 2 101 98

1000 1006 3 101 98

96 17-May-2013 21-May-2013 0 n.d. n.a.

1000 1025 2 103 99

1000 1037 3 104 101

1 Samples were stored in the freezer (≤ -15°C) until the day of analysis. 2 Sample A

3 Sample B

n.d. Not detected. 

n.a. Not applicable.

 

Final Report