REPORT 

Study Title: 

TOXICITY OF 

DEAIACID ANHYDRIDE REACTION PRODUCT 

TO SCENEDESMUS SUBSPICATUS 

IN A 72-HOUR ALGAL GROWTH INHIBITION TEST 

Data Requirements I Test Guidelines: 

OECD No. 201 

EU Commission Directive 92/69/EEC, C.3 

Study Director: 

Roger B~Uscher 

Study Completion Date: 

November 19, 2004 

Test Facility: 

RCC Ltd 

Environmental Chemistry & 

Pharmanalytics 

CH-4452 ltingen I Switzerland 

Sponsor: 

Rockwool International AIS 

Hovedgaden 584 

DK-2640 Hedehusene 

Denmark 

Identification: 

RCC Study Number: 

Page 1 of 59 

848634 
RCC Study Number 848634 

DENAcid anhydride reaction product 

GLP CERTIFICATE 

REPORT 

The Swiss GLP Monitoring Authorities 

Swiss Federal 

Office of 

Public Health 

Swiss Agency for the 

Environment. Forests 

and Landscape 

~ Swissmedic 

r . Swiss Agency for SWlSsrnedlC TherapeuUcProducts 

Statement of GLP Compliance 

It is hereby confirmed that 

during the period of 

the following Test Facilities of 

November 18 - 22, 2002 

RCC Ltd 

4452 ltingen 

Switzerland 

were inspected by the Federal Office of Public Health, the Swiss Agency for Therapeutic 

Products and the Swiss Agency for the Environment, Forests and Landscape with respect to 

the compliance with the Swiss legislation on Good Laboratory Practice. 

Test Facilities 

-Toxicology 

- Environmental Chemistry and 

Pharmanalytics 

Areas of expertise * 

TOX, ACC, MUT, 

OTH (Safety Pharmacology) 

ACC, ECT, ENF, EMN, PCT, 

RES, OTH (Animal metabolism) 

The inspections were performed in agreement with the OECD Guidelines for National GLP 

Inspections and Audits. It was found that the aforementioned test facilities were operating in 

compliance with the Swiss Ordinance relating to Good Laboratory Practice [RS 813.016.5] at 

the time they were inspected. 

Bern, March 2003 

Federal Office of Public Health 

The Director 

Prof. Th. Zeltner 

• TOX =Toxicology: ACC =Analytical and Clinical Chemislly: ECT =Environmental toxicity on aquatic and terresbial 

organisms ; ENF = Behaviour In water, soli and air. Bioaccumulalion ; EMN = Studies on effecls on mesocosms and natural 

ecosystems; MUT = Mutagenicity; PCT = Physical-chemical testing ; RES = Residue studies ; OTH "' Other, to be speclfied. 

Page2 
RCC Study Number 848634 

DEA/Acid anhydride reaction product 

GOOD LABORATORY PRACTICE 

REPORT Page 3 

STATEMENT OF COMPLIANCE 

RCC Study Number: 

Test Item: 

Study Director: 

Study Title: 

848634 

DENAcid anhydride reaction product 

Mr. Roger Bi:itscher 

Toxicity of DENAcid anhydride reaction product 

to Scenedesmus subspicatus 

in a 72-hour algal growth inhibition test 

This study (with the exception of the pre-experiments as mentioned in the report and the 

analytical standards as mentioned in the analytical phase report) has been performed in 

compliance with the Swiss Ordinance relating to Good Laboratory Practice, adopted February, 

2"d, 2000 [RS 813.016.5]. This Ordinance is based on the OECD Principles of Good 

Laboratory Practice, as revised in 1997 and adopted November 26th, 1997 by decision of the 

OECD Council [C(97) 186/Final]. 

There were no circumstances that may have affected the quality or integrity of the data. 

Study Director: Mr. Roger Bi:itscher 

Date: J/oll'lAMbJJ )'J 2oolf I 
RCC Study Number 848634 

DEA/Acid anhydride reaction product 

SIGNATURES 

Study Director: 

Management: 

REPORT 

Mr. Roger B~Uscher 

Date: 

Dr. Uwe Morgenroth 

Page4 

;It; 2oo~ f 
RCC Study Number 848634 

DEA/Acid anhydride reaction product 

QUALITY ASSURANCE 

REPORT Page 5 

RCC Ltd, Environmental Chemistry & Pharmanalytics, CH-4452 ltingen I Switzerland 

STATEMENT 

RCC Study Number: 

Test Item: 

Study Director: 

Study Title: 

848634 

DEA/Acid anhydride reaction product 

Mr. Roger Batscher 

Toxicity of DEA/Acid anhydride reaction product 

to Scenedesmus subspicatus 

in a 72-hour algal growth inhibition test 

The general facilities and activities are inspected periodically and the results are reported to, 

the responsible person and the management. 

Study procedures (with the exception of the pre-experiments as mentioned in the report) were 

periodically inspected. The study plan and this report were audited by the Quality Assurance. 

The dates are given below. 

Dates and Types of QA Inspections Dates of Reports 

to the Study Director 

and to the Management 

December 12, 2003 Study plan December 12, 2003 

July 16, 2004 Process based (analytical work) July 16, 2004 

August30, 2004 Report September 01, 2004 

September 01, 2004 

This statement also confirms that this final report reflects the raw data. 

Quality Assurance: FMrs. Andrea Gildel 
RCC Study Number 848634 

DEA/Acid anhydride reaction product 

CONTENTS 

REPORT Page6 

GLP CERTIFICATE ......................................................................................................... 2 

STATEMENT OF COMPLIANCE .................................................................................... 3 

SIGNATURES ................................................................................................................. 4 

QUALITY ASSURANCE .................................................................................................. 5 

CONTENTS ..................................................................................................................... 6 

CONTENTS CONTINUED (TABLES & FIGURES) .......................................................... ? 

PREFACE ....................................................................................................................... 8 

DATA REQUIREMENTS I TEST GUIDELINES ............................................................... 9 

SUMMARY OF STUDY PLAN AMENDMENTS ............................................................... 9 

SUMMARY .................................................................................................................... 10 

1 PURPOSE ............................................................................................................ 11 

2 MATERIALS AND METHODS .............................................................................. 11 

2.1 DEFINITIONS ............................................................................................. 11 

2.2 TEST ITEM ................................................................................................. 12 

2.3 ANALYTICAL STANDARD .......................................................................... 12 

2.4 TEST SYSTEM ........................................................................... , ................ 12 

2.5 STUDY DESIGN ......................................................................................... 13 

2.5.1 Experimental Conditions ................................................................ 13 

2.5.2 Preparation of Test Solutions and Concentrations ......................... 14 

2.6 EVALUATIONS ........................................................................................... 15 

2.6.1 Counting and Examination of the Algal Cells ................................. 15 

2.6.2 Determination of the Algal Growth Inhibition and the EC 

Values ........................................................................................... 15 

2.6.3 Water Quality Criteria .................................................................... 16 

2.6.4 Analysis of the Test Item Concentrations ....................................... 16 

3 RESULTS AND DISCUSSION ............................................................................. 18 

4 TABLES ............................................................................................................... 20 

5 FIGURE ................................................................................................................ 23 

6 REFERENCES ..................................................................................................... 24 

ATTACHMENT .............................................................................................................. 25 

ANALYTICAL PHASE REPORT (Pages 1 to 34) 
RCC Study Number 848634 

DENAcid anhydride reaction product 

REPORT 

CONTENTS CONTINUED (TABLES & FIGURES) 

Page 7 

Table 1: Algal cell densities during the test period of 72 hours ................................ 20 

Table 2: Areas under the growth curves (AUG) and percentage inhibition 

of AUG (lAue) during the test period ........................................................... 21 

Table 3: Growth rates (r) and percentage inhibition of r (lr) 

during the test period ................................................................................ 21 

Table 4: Appearance of the test medium ................................................................ 22 

Table 5: pH values in the test medium and in the control ........................................ 22 

Table 6: Water temperature during the test period .................................................. 22 

Figure 1: Growth curves of Scenedesmus subspicatus, incubated for 72 hours ....... 23 
RCC Study Number848634 

DEA/Acid anhydride reaction product 

PREFACE 

GENERAL 

Study Title: 

Sponsor: 

Study Monitor: 

Test Facility: 

Analytical Phase No.: 

RESPONSIBILITIES 

Study Director: 

Deputy Study Director: 

Principal Investigator for 

Analytical Phase: 

Technical Coordinator: 

Head of RCC Quality Assurance: 

SCHEDULE 

Experimental Starting Date: 

Experimental Completion Date: 

Study Completion Date: 

ARCHIVING 

REPORT Page 8 

Toxicity of DEA/Acid anhydride reaction product 

to Scenedesmus subspicatus 

in a 72-hour algal growth inhibition test 

Rockwool International A/S 

Hovedgaden 584 

DK-2640 Hedehusene 

Denmark 

Mrs. Anne Jensen (until end of May 2004) 

Mrs. Lea Tobiassen, DVM (from June 01, 2004) 

Danish Toxicology Centre 

Kogle Aile 2 

DK-2970 H0RSHOLM 

Denmark 

RCC Ltd 

Environmental Chemistry & Pharmanalytics 

Zelgliweg 1 

CH-4452 ltingen I Switzerland 

848635 

Mr. Roger B~itscher 

Dr. Birgit Seyfried 

Dr. Tobias Schoop 

Mrs. Helene Eckenstein 

Mrs. Iris Wuthrich 

June 04, 2004 

June 21, 2004 

November 19, 2004 

RCC Ltd, CH-4452 ltingen, Switzerland will retain the study plan, raw data, a sample of the 

test item and the final report of the present study for at least ten years. No data will be 

discarded without the Sponsor's consent. 
RCC Study Number 848634 

DENAcid anhydride reaction product 

1 PURPOSE 

REPORT Page 11 

The purpose of this test was to determine the inhibitory effect of the test item DENAcid 

anhydride reaction product on the growth of the freshwater green algal species Scenedesmus 

subspicatus. Exponentially growing cultures of this algal species were exposed to a single test 

concentration of nominal 100 mg/L under defined conditions (limit test). The limit test was 

based on a range-finding test and was performed in compliance with the EU Commission 

Directive 92/69/EEC to demonstrate that the test item has no toxic effect on Scenedesmus 

subspicatus up to at least this concentration. 

A static, non-renewal exposure system was used. The inhibition of growth in relation to control 

cultures was determined over a test-period of 72 hours and, thus, over several algal 

generations. 

The test method and the test species are recommended by the test guidelines of OECD and 

EU. 

2 MATERIALS AND METHODS 

2.1 DEFINITIONS 

Cell density: 

Growth: 

Biomass (b): 

Growth rate (r): 

NOEC (No Observed 

£_ffect Concentration): 

LOEC (1_owest Observed 

S.ffect Concentration): 

the number of cells per ml 

the increase of cell density over the test period 

the actual number of cells per volume of medium (cells/ml) 

calculated as the g_rea ynder the growth _gurve (AUC) 

the increase of cell density per unit time 

the calculated concentration of test item that results in an x% 

reduction of biomass (b) relative to the control 

the calculated concentration of test item that results in an x% 

reduction of growth rate (r) relative to the control 

the highest test concentration at which no significant inhibition 

of growth is observed relative to the control 

the lowest test concentration at which a significant inhibition of 

growth is observed relative to the control 
RCC Study Number 848634 

DEA/Acid anhydride reaction product 

2.5 STUDY DESIGN 

2.5.1 Experimental Conditions 

REPORT Page 13 

The algae were cultivated and tested in synthetic test water, prepared according to the test 

guidelines. Analytical grade salts were dissolved in sterile purified water to obtain the following 

final nominal concentrations: 

Macro-nutrients: 

Trace elements: 

NaHC03 

CaCb x 2 H20 

NH4CI 

MgS04 x 7 HzO 

MgCI2 x 6 HzO 

KHzP04 

Na2EDTA x 2 H20 

FeCb x 6 HzO 

MnCI2 x 4 HzO 

H3B03 

NazM004 x 2 HzO 

ZnCI2 

CoCb x 6 H20 

CuCI2 x 2 HzO 

50.0 mg/L 

18.0 mg/L 

15.0 mg/L 

15.0 mg/L 

12.0 mg/L 

1.6 mg/L 

100.0 !Jg/L 

80.0 !Jg/L 

415.0 !Jg/L 

185.0 !Jg/L 

7.0 !Jg/L 

3.0 !Jg/L 

1.5 !Jg/L 

0.01 !Jg/L 

Calculated water hardness of the test water: 0.24 mmoi/L (= 24 mg/L as CaC03). 

The test was started (0 hours) by inoculation of 10,000 algal cells per mL test medium. The 

algal cells were taken from an exponentially growing pre-culture which was set up four days 

prior to the test under the same conditions as in the test. One day before the start of the test, 

the pre-culture was diluted threefold to keep the algae in exponential growth. 

The test design included three replicates of the test concentration and six replicates of the 

control. Volumes of 15 mL algal suspension for each replicate were continuously stirred by 

magnetic stirrers in 50 mL Erlenmeyer flasks. 

The flasks were covered with glass dishes. They were incubated in a temperature controlled 

water bath at a temperature between 22 and 23 oc (Table 6) and continuously illuminated at a 

measured light intensity of about 8500 Lux (mean value), range: 8010 to 8910 Lux (minimum 

and maximum value of measurements at nine places distributed over the experimental area at 

the surface of the test media). This illumination was achieved by fluorescent tubes (Philips 

TLD 36W/840), installed above the test flasks. The test vessels were labeled with the RCC 

study number and all necessary additional information to assure unmistakable identification. 

The test duration was 72 hours. 

For evaluation of the algal quality and the experimental conditions, potassium dichromate is 

tested as a positive control at least once a year to demonstrate satisfactory conditions of the 

test. The latest result of the positive control test in 2003 (72-hour EC50 for the growth rate: 

0.69 mg/L, RCC Study 849621) showed that the toxic performance was valid and within the 

historical range of the RCC laboratory (from 1996 to 2003: 72-hour EC50: 0.44- 1.16 mg/L). 
RCC Study Number 848634 

DENAcid anhydride reaction product 

REPORT 

2.5.2 Preparation of Test Solutions and Concentrations 

Page 14 

A limit test was performed in accordance with the EU Commission Directive 92/69/EEC to 

demonstrate that the test item has no toxic effect on the algae up to and including the nominal 

test item concentration of 100 mg/L. Thus, a single test item concentration of nominal 

100 mg/L was tested. Additionally, a control was tested in parallel (test water without addition 

of the test item). 

The test medium was prepared immediately before the start of the test as follows: The amount 

of 60.3 mg of the test item was mixed into 600 ml test water as homogeneously as possible 

using ultrasonic treatment for 15 minutes and intense stirring for 15 minutes at room 

temperature. 

The actual concentration of the test item (i.e. of the different components of the test item) in 

the test medium was analytically determined (see Section 2.6.4). 

The limit test was based on the results of a range-finding test and on the results of a preexperiment to determine the solubility of the test item (without GLP). However, concentrations 

above nominal 100 mg/L were not tested according to the EU Commission Directive 

92/69/EEC. 
RCC Study Number 848634 

DENAcid anhydride reaction product 

2.6 EVALUATIONS 

REPORT 

2.6.1 Counting and Examination of the Algal Cells 

Page 15 

Small volumes of the test medium and the control (1.0 ml) were taken out of all test flasks 

after 24, 48, and 72 hours of exposure, and were not replaced. The algal cell densities in the 

samples were determined by counting with an electronic particle counter (Coulter Counter®, 

Model ZM), with at least two measurements per sample. 

In addition, after 72 hours exposure, a sample was taken from the control and from the single 

test concentration. The shape of the algal cells was examined microscopically in these 

samples. 

2.6.2 Determination of the Algal Growth Inhibition and the EC Values 

Inhibition of algal growth was determined from: 

a) the area under the growth curves AUC (= biomass, b) 

b) the specific growth rates (r) for exponentially growing cultures 

using the following equations: 

a) Area under the growth curve (AUC): 

where: No 

N1 

Nz 

Nn 

t1 

tz 

tn 

= nominal number of cells/ml at time t0 (start of the test) 

= mean measured number of cells/ml at time t1 

= mean measured number of cells/ml at time t2 

= mean measured number of cells/ml at time tn 

= time of first measurement after start of the test 

= time of second measurement after start of the test 

= time of nth measurement after start of the test 

Percentage inhibition of AUC (lAue): 

I - AUCC - AUCt 1 QQOI AUC- . to 

AUCC 

where: AUCc = mean area under the control growth curve 

AUCt = mean area under the growth curve at treatment t 
RCC Study Number 848634 

DEA/Acid anhydride reaction product 

REPORT Page 17 

Duplicate samples per sampling date were taken for each analysis, resulting in a total number 

of six samples per sampling date from 'the test medium and the control. 

The samples for analysis by method 1 and 3 were analyzed directly after the sampling without 

prior storage. The samples for analysis by method 2 were deep-frozen at approximately -20 oc 

immediately after sampling and were kept deep-frozen until analysis. The stability of the 

components to be analyzed by method 2 under the storage conditions was confirmed in a preexperiment (without GLP). 

The components of the test item were analyzed in all duplicate test medium samples. From 

the control samples, one of the duplicate samples was analyzed per sampling date. 

The analytical procedures and the results are described in the attached analytical phase 

report. 
RCC Study Number 848634 

DENAcid anhydride reaction product 

REPORT Page 19 

No remarkable observations were ma9e concerning the appearance of the test medium. The 

test medium appeared to be a clear solution throughout the test duration (Table 4). 

At the start of the test, the pH values in the test medium and the control were 7. 7 and 8.1, 

respectively. At the end of the test, pH values of the test medium and the control were 8.2 and 

8.4, respectively {Table 5). The water temperature ranged from 22 to 23 oc (Table 6). 
RCC Study Number 848634 

DENAcid anhydride reaction product 

REPORT Page 20 

4 TABLES 

The tabulated values represent rounded results obtained by calculation using the exact raw data. 

Table 1: Algal cell densities during the test period of 72 hours 

Nominal Flask Density of algal cells (cell number x 1 0,000/ml) after 

test item No. 

concentration 

(mg/L) 24 h 

1 3.3 3.1 11.5 

Control 2 2.7 2.5 9.5 

3 2.9 3.0 11.5 

4 2.8 2.7 10.5 

5 2.8 2.7 9.5 

6 3.5 3.3 9.3 

m 2.94 

s 0.31 

n 6 

1 2.9 3.2 11.0 

100 2 2.5 2.4 10.5 

3 2.8 3.0 11.0 

m 2.80 

s 0.31 

n 3 

m: mean value; s: standard deviation; n: number of flasks 

At the start, 10,000 algal cells/ml were incubated. 

48 h 72 h 

10.8 69.7 

9.7 49.1 

11.4 71.4 

10.2 58.6 

9.5 49.2 

8.7 44.1 

10.18 56.99 

0.98 11.46 

6 6 

11.0 64.2 

10.1 60.5 

11.1 60.2 

10.78 61.88 

0.42 2.46 

3 3 

69.8 

49.4 

71.1 

58.0 

50.0 

43.5 

65.2 

61.1 

60.1 
RCC Study Number 848634 

DEA/Acid anhydride reaction product 

REPORT 

Table 2: Areas under the growth curves (AUC) and percentage inhibition 

of AUC (lAue) during the test period 

Page 21 

Nominal Areas under the growth curves (AUC) and % inhibition of AUC 

test item 

concentration 0-24 h 0-48h 0-72 h 

(mg/L) AUC lAue(%) AUC lAue(%) AUC lAue(%) 

Control 23 0.0 157 0.0 939 0.0 

100 22 7.3 161 -2.5 1009 -7.4 

AUC x 10,000 

- % inhibition: increase in growth relative to that of control 

Table 3: Growth rates (r) and percentage inhibition of r (lr) during the test period 

Nominal Growth rate rand % inhibition of r 

test item 

concentration 0-24 h 0-48 h 0-72 h 

(mg/L) r (1/day) lr (%) r (1/day) lr (%) r (1/day) lr (%) 

Control 1.07 0.0 1.16 0.0 1.34 0.0 

100 1.03 4.6 1.19 -2.6 1.37 -2.4 

- % inhibition: increase in growth relative to that of control 
RCC Study Number 848634 

DEA/Acid anhydride reaction product 

REPORT 

Table 4: Appearance of the test me,~ium 

Abbreviations: 

0: no remarkable observations, clear test medium 

1: homogeneous dispersion in the water, turbidity observable 

2: noticeable turbidity caused by the test item 

3: noticeable coloration caused by the test item 

4: inhomogeneous dispersion of the test item 

5: precipitation of the test item 

6: test item at the surface 

7: test item lying at the bottom of the flask 

Nominal test item Exposure time concentration 

(mg/L) Oh 24 h 48 h 

100 0 0 0 

Table 5: pH values in the test medium and in the control 

Nominal test item pH values 

concentration 

(mg/L) Start End 

Control 8.1 8.4 

100 7.7 8.2 

Table 6: Water temperature during the test period 

Temperature (

0

C} 

Day 0 (Start) 22 

Day 1 23 

Day2 22 

Day 3 (End) 23 

Page22 

72h 

0 
RCC Study Number 848634 

DENAcid anhydride reaction product 

5 FIGURE 

REPORT 

Figure 1: Growth curves of Scenedesmus subspicatus, incubated for 72 hours 

100-.-----------------------------------------. 

::::J 

E 

""" --

0 

>< 

!!2 

Qj 

.s. 

>-

"iii 

- 10 c 

Q) 

"0 

Qj 

(.) 

c 

m 

Q) 

E 

1 

0 24 48 72 

test period (hours) 

··•· Control -D- 100 mg/L 

Page 23 
RCC Study Number 848634 

DENAcid anhydride reaction product 

6 REFERENCES 

REPORT 

1 AN, S.S., FRIEDL, T., HEGEWALD, E. (1999): 

Page 24 

Phylogenetic Relationships of Scenedesmus and Scenedesmus-like Coccoid Green Algae 

as Inferred from ITS-2 rONA Sequence Comparisons, 

Plant Biology 1, 418-428 

2 HEGEWALD, E. (2000): 

New Combinations in the Genus Oesmodesmus (Chlorophytaceae, Scenedesmaceae) 

Algological Studies 96, 1-18 
RCC Study Number 848634 

DENAcid anhydride reaction product 

REPORT 

ATTACHMENT 

ANALYTICAL PHASE REPORT 

Page 25 
ANALYTICAL PHASE REPORT 

Analytical Phase to: 

TOXICITY OF 

DEAIACID ANHYDRIDE REACTION PRODUCT 

TO SCENEDESMUS SUBSPICATUS 

IN A 72-HOUR ALGAL GROWTH INHIBITION TEST 

Study Director: 

Mr. Roger Batscher 

Principal Investigator Analytical Phase: 

Dr. Tobias Schoop 

Completion Date of the Analytical Phase Report: 

November 19, 2004 

Test Facility: 

RCC Ltd 

Environmental Chemistry & Pharmanalytics 

CH-4452 ltingen/Switzerland 

Identification: 

RCC Study Number: 848634 

RCC Phase Number: 848635 

Page 1 to 34 
RCC Study Number 848634 ANALYTICAL PHASE REPORT 

DENAcid anhydride reaction product 

RCC Phase Number 848635 

GOOD LAB ORA TORY PRACTICE 

STATEMENT OF COMPLIANCE 

RCC Study No.: 

Study Director: 

Analytical Phase Number: 

Test Item: 

Principal Investigator for 

Analytical Phase: 

Analytical Phase to: 

848634 

Mr. Roger B~itscher 

RCC Ltd 

Environmental Chemistry & Pharmanalytics 

848635 

DENAcid anhydride reaction product 

Dr. Tobias Schaap 

Toxicity of DENAcid anhydride reaction product 

to Scenedesmus subspicatus 

in a 72-hour algal growth inhibition test 

Page 2 

The analytical phase (with the exception of the pre-experiments and the standards used for 

identification of the interesting signals as mentioned in the phase report) has been performed 

in compliance with the Swiss Ordinance relating to Good Laboratory Practice, adopted 

February 2nd, 2000 [RS 813.016.5]. This Ordinance is based on the OECD Principles of Good 

Laboratory Practice, as revised in 1997 and adopted November 261

h, 1997 by decision of the 

OECD Council [C(97)186/Final]. 

Principal Investigator for 

Analytical Phase: Dr. Tobias Schaap 

i?;Gn0/ 

···················~···· .. ················· .. 

Date: ;f/vvA-Gv .-A"1 / Q_(f!Y~ 
RCC Study Number 848634 ANALYTICAL PHASE REPORT 

DEA/Acid anhydride reaction product 

RCC Phase Number 848635 

PREFACE 

GENERAL 

Analytical Phase to: 

Sponsor: 

Study Monitor: 

Test Facility: 

RESPONSIBILITIES 

Study Director: 

Principal Investigator 

for analytical phase: 

Technical Coordinators: 

Reporting: 

Head of RCC 

Quality Assurance: 

Toxicity of DEA/Acid anhydride reaction product 

to Scenedesmus subspicatus 

in a 72-hour algal growth inhibition test 

Rockwool International A/S 

Hovedgaden 584 

DK-2640 Hedehusene 

Denmark 

Mrs. Anne Jensen (until end of May 2004) 

Mrs. Lea Tobiassen, DVM (from June 01, 2004) 

Danish Toxicology Centre 

Kogle Aile 2 

DK-2970 H0RSHOLM 

Denmark 

RCC Ltd 

Environmental Chemistry & Pharmanalytics 

Zelgliweg 1 

CH-4452 ltingen/Switzerland 

Mr. Roger Batscher 

Dr. Tobias Schoop 

Mrs. Mandy Broda, Mr. Roman Klaus 

Dr. Tobias Schoop 

Mrs. Iris Wuthrich 

SCHEDULE OF ANALYTICAL PHASE 

Experimental starting date: 

Experimental completion date: 

Completion date of the Analytical Phase Report: 

June 07, 2004 

June 21, 2004 

November 19, 2004 

Page 5