## FINAL REPORT

Study Title



## ACUTE TOXICITY STUDY IN **DAPHNIA MAGNA** WITH

EXP1313100

(STATIC)

Author

Test Facility

WIL Research Europe B.V.

Hambakenwetering 7

5231 DD ‘s

- Hertogenbosch

The Netherlands

Laboratory Project Identification

Project 505715

Substance 205622/A

- Page 1 of 22

-EXP1313100 Project 505715

- Page 2 - 1. CONTENTS

1. CONTENTS

................................................................................................ ................................ 2

2. STATEMENT OF GLP CO MPLIANCE

......................................................................................... 3

3. QUALITY ASSURANCE ST ATEMENT

........................................................................................ 4

4. SUMMARY

................................................................................................ .................................. 5

5. INTRODUCTION

......................................................................................................................... 6

5.1. Preface

................................................................................................ ................................ 6

5.2. Aim of the study

................................................................................................................... 6

5.3. Guideline s

............................................................................................................................ 6

5.4. Storage and retention of records and materials

..................................................................... 6

5.5. Definitions

............................................................................................................................ 7

6. MATERIALS AND METHOD S

..................................................................................................... 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 syst em

......................................................................................................................... 7

6.3. Breeding

.............................................................................................................................. 8

6.4. Preparation of test solutions

................................................................................................ . 9

6.5. C ombined limit/range

- finding 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. Immobility

........................................................................................................................... 11

7.3. Determination of effect concentrations

................................................................................ 11

7.4. Experimental conditions

..................................................................................................... 11

8. CONCLUSION

.......................................................................................................................... 12

TABLES

Table 1 Num ber of introduced daphnids and incidence of immobility

..................................... 11

Table 2 Effect parameters

..................................................................................................... 11

Table 3 pH and oxygen concentrations

................................................................................. 12

APPENDICES

APPENDIX 1 REFERENCE TEST

................................................................................................ . 13

APPENDIX 2 CERTIFICATE OF ANALYSIS

.................................................................................. 14

APPENDIX 3 ANALYTICAL REPORT

............................................................................................ 15EXP1313100 Project 505715

- Page 5 - 4. SUMMARY

Acute Toxicity Study in Daphnia magna with EXP1313100

.

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

, 2004 . In

addition, the procedures were designed to meet the test methods of the Commission Regulation (EC)

No 440/200 8

, Part C.2, 200 8

, the ISO Inter national Standard 6341, 1996 and the OECD series on

testing and assessment number 23, 2000.

The batch of EXP1313100 tested was a brown solid that was not completely soluble in test medium at

the loading rates initially prepared

.

All test solutions were prepared separately. Various loading rates of the test substance were added to

the test medium applying two days of magnetic stirring in order to reach maximum solubility. The

resulting aqueous mixtures were filtered through a 0.45 μm membrane filter where after the clear and

colourless Water Accommodated Fractions (WAFs) were used for testing

.

A combined limit/range

- finding test was performed. Twenty daphnids per group ( four replicate s, five

daphnids per replicate ) were exposed to a WAF prepared at a loading rate of 100 mg/L and to a

control. In addition

, ten daphnids per group (two replicates

, five daphnids per replicate) were exposed

to WAFs prepared at loading rate s of 1.0 and 10 mg/L . The total test period was 48 hours and

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

of the test.

T he initial concentrations in the WAFs prepared at loading rates of 1.0, 10 and 100 mg/L were 0.029,

0.60 and 0.21 mg/L. These concentrations remained stable during the test period (88

- 98% of initial at

the end of the test. The concentrations obtained in the WAFs prepared at loading rates of 10 and

100 mg/l indicated that the maximum solubility in test medium was exceeded at these loading rates.

This conclusion was based on the water solubility of 0.02 mg/L, which was determined in Project

505730.

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

No effect on mobility was recorded at any of the loading rates of EXP1313100 teste d.

The 48h

- EL* 50 was above a loading rate of 100 mg EXP1313100 per litre

. The concentration of

EXP1313100 in the WAF prepared at a loading rate of 100 mg/L was determined to be 0.21 mg/L

based on the initial exposure concentration. This concentration was considered to be above the

maximum solubility of EXP1313100 in the test medium.

*EL = effective loading rate