REPORT 

Study Title 

ACUTE TOXICITY OF 

DEAIACID ANHYDRIDE REACTION PRODUCT 

TO DAPHNIA MAGNA 

IN A 48-HOUR IMMOBILIZATION TEST 

Data Requirements I Test Guidelines: 

OECD No. 202, Part I 

EU Commission Directive 92/69/EEC, C.2 

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 A/S 

Hovedgaden 584 

DK-2640 Hedehusene 

Denmark 

Identification: 

RCC Study Number: 

Page 1 of 52 

848632 
RCC Study Number 848632 

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 SWlSSrnedtc 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 terreslrlal 

organisms ; ENF = Behaviour In water, soil and air. BioaccumulaUon ; EMN = Studies on effects on mesocosms and natural 

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

Page2 
RCC Study Number 848632 

DENAcid anhydride reaction product 

GOOD LABORATORY PRACTICE 

REPORT Page 3 

STATEMENT OF COMPLIANCE 

RCC Study Number: 

Test Item: 

Study Director: 

Study Title: 

848632 

DEA/Acid anhydride reaction product 

Mr. Roger Batscher 

Acute toxicity of DEA/Acid anhydride reaction product 

to Daphnia magna in a 48-hour immobilization 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"ct, 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]. 

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

Study Director: Mr. Roger Batscher 

Date: J;o~reiM/aJ.J .A5, 2.oof(-
RCC Study Number 848632 

DEA/Acid anhydride reaction product 

SIGNATURES 

Study Director: 

Management: 

REPORT Page4 

Mr. Roger Batscher 

Date: JJo lrltM~tJ )~, too~ 

Dr. Uwe Morgenroth 

.. fL ..... . 

Date: 
RCC Study Number 848632 

DEA/Acid anhydride reaction product 

REPORT Page 5 

QUALITY ASSURANCE 

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

STATEMENT 

RCC Study Number: 

Test Item: 

Study Director: 

Study Title: 

848632 

DEA/Acid anhydride reaction product 

Mr. Roger B~itscher 

Acute toxicity of DEA/Acid anhydride reaction product 

to Daphnia magna in a 48-hour immobilization 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 22, 2004 Process based July 22, 2004 

(test system, test conditions) 

October 16/17/18, 2004 Report October 18, 2004 

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

Quality Assurance: ~rs. Andrea Gudel 
RCC Study Number 848632 

DENAcid anhydride reaction product 

CONTENTS 

REPORT Page6 

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

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

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

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

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

CONTENTS CONTINUED (TABLES) .............................................................................. ? 

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 Determination of the Immobility, NOEC, ECO, EC50, 

and EC100 .................................................................................... 15 

2.6.2 Water Quality Criteria .................................................................... 15 

2.6.3 Analysis of the Test Item Concentrations ....................................... 15 

3 RESULTS AND DISCUSSION ............................................................................. 16 

4 TABLES ............................................................................................................... 17 

ATTACHMENT .............................................................................................................. 19 

ANALYTICAL PHASE REPORT (Pages 1 to 33) 
RCC Study Number 848632 

DENAcid anhydride reaction product 

REPORT 

CONTENTS CONTINUED (TABLES) 

Table 1: Effect of DEN Acid anhydride reaction product on the mobility 

Page 7 

of Daphnia magna ....................................................................................... 17 

Table 2: Appearance of the test medium during the test period ................................ 17 

Table 3: Dissolved oxygen concentrations, pH values, and temperature 

in the test medium and the control .............................................................. 18 
RCC Study Number 848632 

DENAcid 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 

Acute toxicity of DEN Acid anhydride reaction 

product to Daphnia magna in a 48-hour 

immobilization test 

Rockwoollnternational NS 

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 

KogleAIIe 2 

DK-2970 H0RSHOLM 

Denmark 

RCC Ltd 

Environmental Chemistry & Pharmanalytics 

Zelgliweg 1 

CH-4452 ltingen I Switzerland 

848633 

Mr. Roger B~itscher 

Dr. Armin Peither 

Dr. Tobias Schoop 

Mr. Frederic Levrechon 

Mrs. Iris Wuthrich 

June 07,2004 

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 848632 

DENAcid anhydride reaction product 

REPORT Page 11 

1 PURPOSE 

The purpose of the 48-hour toxicity test was to evaluate the effect of the test item 

DEA/Acid anhydride reaction product on the mobility of Daphnia magna. Young daphnids 

were exposed in a 48-hour static test to an aqueous test medium containing the test item at 

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

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

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

daphnids up to at least this concentration. The recorded effect was the immobility of the 

daphnids. 

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

EU. 

2 MATERIALS AND METHODS 

2.1 DEFINITIONS 

Immobilization: 

ECO: 

EC50: 

EC100: 

NOEC (No Observed 

,!;ffect Concentration): 

those organisms which are not able to swim within 15 seconds 

after gentle agitation of the test container are considered to be 

immobile 

the highest test concentration without a significant number of 

immobilized test organisms 

the calculated concentration of test item which results in 50% 

immobilization 

the lowest test concentration at which all test organisms are 

immobile 

the highest test concentration at which no significant toxic effect 

on the test organisms is observed 
RCC Study Number 848632 

DEA/Acid anhydride reaction product 

2.5 STUDY DESIGN 

2.5.1 Experimental Conditions 

REPORT Page 13 

The test was performed in 100 ml glass beakers filled with 50 ml of test medium. The 

beakers were covered with glass plates to reduce the loss of water and to avoid the entry of 

dust into the solutions. The test vessels were labeled with the RCC study number and all 

necessary additional information to ensure unmistakable identification. 

At the single test concentration and for the control, 20 daphnids were used divided into four 

replicates of five daphnids each. The daphnids were randomly distributed to the test vessels 

at initiation of the test. The loading rate was less than one daphnia per 5 ml of test solution. 

A static test without test medium renewal was performed. 

Water temperature: 20 - 21 oc during the test period (Table 3). The test was performed in a 

temperature-controlled room (room temperature continuously 

monitored). 

Light conditions: A 16-hour light to 8-hour dark photoperiod (with a 30 minute transition 

period). Light intensity during the light period was between 

approximately 520 and 710 Lux. 

Test duration: 48 hours 

Test water: Reconstituted test water: analytical grade salts were dissolved in purified 

water to obtain the following nominal concentrations: 

CaCI2 x 2H20 

MgS04 x 7H20 

NaHC03 

KCI 

Water Hardness 

Alkalinity 

Ratio of Ca : Mg 

Na:K 

2.0 mmoi/L (= 294 mg/L) 

0.5 mmoi/L (= 123 mg/L) 

0.75 mmoi/L (= 65 mg/L) 

0.075 mmoi/L (= 5.8 mg/L) 

2.5 mmoi/L (= 250 mg/L as CaC03) 

0.8 mmoi/L 

= 4 : 1 (based on molarity) 

= 1 0 : 1 (based on molarity) 

The test water was aerated prior to the start of the study until oxygen 

saturation was reached. During the test period, the test water was not 

aerated. 

Feeding: None 

Positive Control: For evaluation of the quality of the daphnia clone and the experimental 

conditions, potassium dichromate is tested as a positive control at least 

once a year. The latest result of the positive control test in 2004 

(48-hour EC50: 0.83 mg/L, RCC Study no. 852355) showed that the 

toxic performance was valid and within the historical range of the RCC 

laboratory (from 1996 to 2004: 48-hour EC50: 0.55 - 1.1 mg/L). 
RCC Study Number 848632 

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 Daphnia magna 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 by mixing 50.2 mg of 

the test item into 500 ml test water 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 2.6.3). 

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 848632 

DENAcid anhydride reaction product 

4 TABLES 

REPORT Page 17 

Table 1: Effect of DEN Acid anhydride reaction product on the mobility of Daphnia magna 

Nominal No. of Immobilized 

test item daphnids daphnids after 

concentration tested 24 hours 

(mg/L) No. % 

Control 20 0 0 

100 20 0 0 

Table 2: Appearance of the test medium during the test period 

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 test beaker 

Nominal 

test item Exposure time 

concentration 

(mg/L) 0 hours 24 hours 

100 0* 0 

* few fine particles observed 

Immobilized 

daphnids after 

48 hours 

No. % 

0 0 

0 0 

48 hours 

0 
RCC Study Number 848632 

DEA/Acid anhydride reaction product 

REPORT 

Table 3: Dissolved oxygen concent~.ations, pH values, and temperature 

in the test medium and the control 

Nominal 

Page 18 

test item Start (0 hours) End (48 hours) 

concentration 

pH Oxygen Temperature pH Oxygen Temperature 

(mg/L) (mg/L) (OC) (mg/L) (OC) 

Control 7.9 9.2 20 7.9 8.6 21 

100 7.4 9.1 20 7.8 8.6 21 
RCC Study Number 848632 

DENAcid anhydride reaction product 

REPORT 

ATTACHMENT 

ANALYTICAL PHASE REPORT 

Page 19 
ANALYTICAL PHASE REPORT 

Analytical Phase to: 

ACUTE TOXICITY OF 

DEAIACID ANHYDRIDE REACTION PRODUCT 

TO DAPHNIA MAGNA 

IN A 48-HOUR IMMOBILIZATION TEST 

Study Director: 

Mr. Roger B~Hscher 

Principal Investigator Analytical Phase: 

Dr. Tobias Schaap 

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: 848632 

RCC Phase Number: 848633 

Page 1 to 33 
RCC Study Number 848632 ANALYTICAL PHASE REPORT Page2 

DENAcid anhydride reaction product 

RCC Phase Number 848633 

GOOD LABORATORY PRACTICE 

STATEMENT OF COMPLIANCE 

RCC Study No.: 

Study Director: 

Analytical Phase Number: 

Test Item: 

Principal Investigator for 

Analytical Phase: 

Analytical Phase to: 

848632 

Mr. Roger Batscher 

RCC Ltd 

Environmental Chemistry & Pharmanalytics 

848633 

DENAcid anhydride reaction product 

Dr. Tobias Schoop 

Acute toxicity of DEN Acid anhydride reaction product to 

Daphnia magna in a 48-hour immobilization test 

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 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]. 

Principal Investigator for 

Analytical Phase: Dr. Tobias Schoop 

.l.?.-. .. ~ .................... . 

Date: .tf/OVfL-(.)y 41 r ?_roC, 
RCC Study Number 848632 ANALYTICAL PHASE REPORT Page 3 

DEA/Acid anhydride reaction product 

RCC Phase Number 848633 

CONTENTS 

STATEMENT OF COMPLIANCE .................................................................................... 2 

CONTENTS .................................................................................................................... 3 

PREFACE ....................................................................................................................... 5 

GENERAL ....................................................................................................................... 5 

RESPONSIBILITIES ........................................................................................................ 5 

SCHEDULE OF ANALYTICAL PHASE ........................................................................... 5 

1 PURPOSE .............................................................................................................. 6 

2 MATERIALS AND METHODS ................................................................................ 6 

2.1 TEST ITEM ............................................................................................................ 6 

2.2 ANALYTICAL PROCEDURE .................................................................................. 7 

2.2.1 Storage ....................................................................................................... 7 

2.2.2 Reagents and Solvents ............................................................................... 7 

2.2.3 Standard Solutions used for Sample Quantification .................................... 7 

2.2.4 Preparation of Spiked Test Water Samples ................................................ 7 

2.2.5 Analysis of Samples .................................................................................... 8 

2.3 MEASUREMENT CONDITIONS ............................................................................ 8 

2.3.1 Method 1 (diethanolamine) ......................................................................... 8 

2.3.2 Method 2 (dimers, trimers and polymers) .................................................... 9 

2.3.3 Method 3 (tetrahydrophthalic acid and trimellitic acid) ................................. 9 

2.4 EVALUATION OF RESULTS ............................................................................... 10 

2.4.1 DEA/Acid anhydride reaction product on the base of diethanolamine ....... 10 

2.4.2 DEA/Acid anhydride reaction product on the base of dimers, trimers 

and polymers ............................................................................................ 11 

2.4.3 DEA/Acid anhydride reaction product on the base of 

tetrahydrophthalic acid .............................................................................. 12 

2.4.4 DEA/Acid anhydride reaction product on the base of trimellitic acid .......... 13 

3 RESULTS AND DISCUSSION ............................................................................. 14 

3.1 METHOD 1 (DIETHANOLAMINE) ........................................................................ 14 

3.2 METHOD 2 (DIMERS/TRIMERS AND POLYMERS) ............................................ 14 

3.3 METHOD 3 (TETRAHYDROPHTHALIC ACID AND TRIMELLITIC ACID) ............ 15 

4 TABLES ............................................................................................................... 17 

Table 1: Example of calibration data of test item standards for diethanolamine ........ 17 

Table 2: Calibration data of test item standards for dimers, trimers and 

polymers ..................................................................................................... 18 

Table 3: Example of calibration data of test item standards for 

tetrahydrophthalic acid ( cis-4-cyclohexene-1 ,2-dicarbonic acid) .................. 19 

Table 4: Example of calibration data of test item standards for trimellitic acid 

(1 ,2,4-Benzenetricarboxylic acid) ................................................................ 20 

Table 5: Results obtained for the concentrations of the test item in the test 

medium on the base of diethanolamine ....................................................... 21 
RCC Study Number 848632 ANALYTICAL PHASE REPORT 

DEA/Acid anhydride reaction product 

RCC Phase Number 848633 

Table 6: Results obtained for the concentrations of the test item in the test 

medium on the base of dihlers, trimers and polymers ................................. 22 

Table 7: Results obtained for the concentrations of the test item in the test 

medium on the base of tetrahydrophthalic acid (cis-4-cyclohexene-1 ,2-

dicarbonic acid) ........................................................................................... 23 

Page4 

Table 8: Results obtained for the concentrations of the test item in the test 

medium on the base of trimellitic acid (1 ,2,4-Benzenetricarboxylic 

acid) ............................................................................................................ 24 

5 FIGURES ............................................................................................................. 25 

5.1 Method 1 (diethanolamine) ....................................................................... 25 

Figure 1: HPLC-chromatogram of standard solution (low-level) ..................................... 25 

Figure 2: HPLC-chromatogram of standard solution (high-level) ................................... 25 

Figure 3: HPLC-chromatogram of biological control sample .......................................... 26 

Figure 4: HPLC-chromatogram of spiked test water sample .......................................... 26 

Figure 5: HPLC-chromatogram of non-aged treatment sample ..................................... 27 

Figure 6: HPLC-chromatogram of aged treatment sample ............................................ 27 

5.2 Method 2 (Dimers/trimers and Polymers) .................................................. 28 

Figure 7: HPLC-chromatogram of standard solution (low-level) ..................................... 28 

Figure 8: HPLC-chromatogram of standard solution (high-level) ................................... 28 

Figure 9: HPLC-chromatogram of biological control sample .......................................... 29 

Figure 10: HPLC-chromatogram of spiked test water sample ........................................ 29 

Figure 11: HPLC-chromatogram ofnon-aged treatment sample ................................... 30 

Figure 12: HPLC-chromatogram of aged treatment sample .......................................... 30 

5.3 Method 3 (tetrahydrophthalic acid and trimellitic acid) ............................... 31 

Figure 13: HPLC-chromatogram of standard solution (low-level) ................................... 31 

Figure 14: HPLC-chromatogram of standard solution (high-level) ................................. 31 

Figure 15: HPLC-chromatogram of biological control sample ........................................ 32 

Figure 16: HPLC-chromatogram of spiked test water sample ........................................ 32 

Figure 17: HPLC-chromatogram of non-aged treatment sample ................................... 33 

Figure 18: HPLC-chromatogram of aged treatment sample .......................................... 33 
RCC Study Number 848632 ANALYTICAL PHASE REPORT Page 5 

DEA/Acid anhydride reaction product 

RCC Phase Number 848633 

PREFACE 

GENERAL 

Analytical Phase to: Acute toxicity of DEA/Acid anhydride reaction product to 

Daphnia magna in a 48-hour immobilization test 

Sponsor: 

Study Monitor: 

Test Facility: 

RESPONSIBILITIES 

Study Director: 

Principal Investigator 

for analytical phase: 

Technical Coordinator: 

Reporting: 

Head of RCC 

Quality Assurance: 

Rockwoollnternational 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 B~itscher 

Dr. Tobias Schoop 

Mrs. Mandy Broda 

Dr. Tobias Schoop 

Mrs. Iris Wuthrich 

SCHEDULE OF ANALYTICAL PHASE 

Experimental starting date: 

Experimental completion date: 

Completion date of the Analytical Phase Report: 

June 08, 2004 

June 21, 2004 

November 19, 2004 
RCC Study Number 848632 ANALYTICAL PHASE REPORT Page 16 

DEA/Acid anhydride reaction product 

RCC Phase Number 848633 

Concurrent with the sample analysis, recoveries of spiked test water samples at relevant 

concentrations (101 mg test item/L) were performed in duplicate. The average concentration 

was found to be 97% of the spiked values (n = 4). No correction for any losses during the 

analytical procedure was made. 

The routine limit of quantification (LOQ) for the test item in test medium was derived from the 

lowest standard solution, which fits into the calibration curve: the value is 10.9 mg/L. With a 

factor of 1.111 from sample dilution a LOQ of approximately 10 mg/L of the test item results. 

The limit of detection (LOD) was calculated to be at 2 mg/L of the test item (1 fifth of the 

LOQ). 

The biological control samples and an analyzed analytical blank (test water) did not affect the 

HPLC-chromatogram at the retention time of the test item. 

The average concentrations found in the treatment samples were 98 % of the nominal 

concentrations at test start and 99 % of the nominal concentrations at test end. 

It can be seen from Table 8 that trimellitic acid was stable during the performance of the 

biological test. 

Typical HPLC chromatograms are shown in the attached Figure 13 to Figure 18.