Document O112BoYBRO65zbGGjworQoMVM

M s. 2 6 - 3 2 7 7 M 1 2 6 -& 7 7 DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Effects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to T r a u n s p u r g e r et al. (19 97 ) Sponsor Asahi Glass C la r ia n t Daikin In d u str ies Ltd D upont Author Gunda W inkelm ann Test Facility D r.U .N o ack-La bo ra to rium F r A n g e w a n d te B io lo g ie Kthe-Paulus-Strae 1 D-31157 Sarstedt Laboratory Prgiect ID Project-No. Study-No. Study-No.(Study pi, Study Com pleted On 2003-04-14 Report Page 1 of 20 ftnrnnanw SanWaed. Does not contain T S C A CB DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE 1Effects on the D evelopm ent of Caenorhabditis elegans in a W ater-S edim en t System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) Page 2 of 20 MProject-No| Study-No. Title Guideline Test Item Statement of GLP Compliance Effects on the Development ot Caenorhabditis elegans in a W aterSediment System T r aunspurg er et al. (1997) Test Facility D R .U .N O A C K -LA B O R A TO R IU M FOR ANGEW ANDTE BIO LO G IE .Phone: (+49) 5 0 6 6 / 706 70, Fax: (+49) 50*66 / 706 789 Deviations from GLP Principles None W e declare that this study was conducted and reported in compliance with the actual OECD principles of Good Laboratory Practice and the national GLP regulations as specified in the law in force, except deviations mentioned above. 2003-04-14 (Date) (Gunda Winkelmann, Study Director) 2003-04-14 (Date) (Dr. Udo Noack,, Head of Testing Facility) Company Sanfflzed. Does not contain T SC A CBI DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Effects on the D evelopm ent of Caenorhabditis elegans in a W ater-S edim en t System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) Page 3 of 20 ' Project-N Study-No Statement of the Quality Assurance Unit Title Guideline Test Item Effects on the Development of Caenorhabditis elegans in a W a te l Sediment System T r a u n s p u r g e r et al. (1 9 9 7 ) Study Director Gunda Winkelmann T he study was verified as follows: inspection study plan process based report dates 2002-11-18 2002-11-21 2002-11-26 2003-01-13 2003-01-14 2003-01-15 2003-03-06 2003-04-10 date of report 2002-11-18 2002-11-21 2002-11-26 2003-01-14 2003-01-14 2003-01-15 2003-03-06 2003-04-10 The reported results accurately and completely reflect the raw data of the study. Also methods, procedures, and observations are accurately and completely described in the report. T h e accordance of the study with its study plan and the principles of Good Laboratory Practice is guaranteed. 2003-04-14 Gudrun MOhrmann-Kalabokidis Company Sanitized. Does not contain TSCA CBI DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE .R e p o rt Effects on the Developm ent of Caenorhabditis elegans in a W ater-S edim en t System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) Page 4 of 20 Project-N Study-No. Personnel Involved Study Director: Gunda Winkelmann (Engineer of Horticulture) Deputy: Dirk Scheerbaum (Biologist) Technical Staff: Karin Ruthenberg Quality Assurance Unit: Gudrun Mhrmann-Kalabokidis (Biologist) Deputy: Susanne Becker (Biologist) Director of the Testing Facility: Dr. Udo Noack (Biologist) Company Sanitized. Does not contain T SCA CBf DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report effects on the Developm ent of Caenorhabditis elegans in a W ater-S edim en t System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) Page 5 of 20 K ro je c t-N o filfH H S S liid y -N o J p g J l I Table of Content Title Page Statement of GLP Compliance Statement of Quality Assurance Unit Personnel Involved Table of Content List of Tables 1 Summary 2 Characterisation Data of the Test Item 3 Method 4 Results 4.1 Preliminary Test 4.2 Definitive Test 5 Validity Criteria 6 Conclusions 7 Literature / References 8 Certificate of Test Item Analysis 9 Certificate of D r .U.Noack-Laboratorium Page 1 2 3 4 5 6 7 8 9 15 15 16 18 18 18 19 20 XSCA CBl iftized. Does notoontein Company Sam DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE 'E ffects on the Developm ent of Caenorhabditis elegans & in a W ater-S edim en t System acc. to T r a u n s p u r g e r et al. (1997) Page 6 of 20 JSBSLProject Study-N List o f Tables Table 1: Table 2: Table 3: Table 4: Table 5: Growth and Reproduction of Caenorhabditis elegans (72 h) Growth and Reproduction of Caenorhabditis elegans (72 h) Bodylength [pm] after 72 hours Number of eggs after 72 hours Growth, Number of eggs and Fertility after 72 hours Page 7 15 16 16 17 S - r f M 0 M iw f < " TS C * C V DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report Effects on the Developm ent of Caenorhabditis elegans in a W ate r-S e d im e n t System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) Page 7 of 20 Project-N Study-No. 1 Summary The effects of (batch P00/001) on the developmlflt of the free living soil nematode Caenorhabditis elegans were determined in a water-sediment system. The study was carried out according to T r a u n s p u r g e r et al. (1997). Test duration was 72 hours. The study was conducted from 2002-11-22 to 2002-11-25 at D r .U .N o a c k - La b o r a t o r iu m FOR A n g e w a n d t e Bio lo g ie , Sarstedt, Germany. A limit test was conducted with a limit concentration of 1000 mg/kg selected on the basis of a preliminary test. Fifty nematodes were exposed to the limit test concentration. Thirty nematodes were tested in an untreated sediment control. A solvent control (fert.-methyl-butyl-ether, MTBE) and a liquid control were also tested with 2 replicates (10 nematodes per replicate) for each control treatment. The effects of the test ite summarised in Table 1. Table 1: Growth and Reproduction of Caenorhabditis elegans (72 h) Test item Test Object Exposure Caenorhabditis elegans Artificial sediment Liquid Control Sediment Control Survival of nematodes [%] 100 - Bodylength [pm] SD 985.7 8 4 .5 Number of eggs per nematode SD .- 3.43 3 .6 7 Solvent Control - 911.5 2 3 .3 6.00 8 .1 3 {mg producl/kg] 1000 - 901.4 5 9 .0 ** SD = Standard deviation * = Percentage of nematodes with > 1 egg ** = No significant difference compared to the pooled control = Not determined 2.98 3.05** I ^0 Gravid w orm s* [%] - 90.0 0.0 90.0 0 .0 92.0 9 .1 0 Company S anitized ;<yf esKtti fS C M C m DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE fects on the Developm ent of Caenorhabditis elegans in a W ater-S edim en t System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) Page 8 of 20 MProject-No Study-No. Appearance W ater solubility Stability in water Density Yellowish solid wax 134 pg/L at 12 degree C, Std 29 pg/L, n=8 (DuPont, 2002) 137 pg/L at 25 degree C, Std 53 pg/L, n=17 (DuPont, 2002) Stable at test conditions Ca. 1.7g/cm3 Expiry date Recommended storage Storage at testing facility Retention Identification parameter at testing facility Unlimited if stored under the recommended conditions. Room temperature (20 C), protected from moisture Room temperature, protected from moisture and light At least 1 g has been retained. Name, batch number and colour The test item and the information concerning the test item were provided by the sponsor. not contain f SCA CBl Sanfttee d - Does C o m p ly DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE ^Effects on the Developm ent of Caenorhabditis elegans in a W ater-S edim en t System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) Page 9 of 20 Project-No Study-No. 3 Method TEST GUIDELINE TYPE AND PURPOSE OF THE STUDY T r a u n spu r g er et al. (1997): "Ecotoxicological assessment o f aquatic sediments with Caenorhabditis elegans (nematoda) - A method for testing in liquid medium and whole sediment samples." 1Determination of the potential impact of the free livin^olm em atode Caenorhabditis elegans. TEST SYSTEM Reason for the selection Source Food medium Feeding Food Caenorhabditis elegans (nematoda) N2, var. Bristol, wild type lab strain According to the literature (T r a u n s p u r g e r et al., 1997) Caenorhabditis elegans is suitable for this kind of study. Breeding was performed in the test facility at 20 2 C in the dark. Origin: C a en o r h a b d itis G e n e t ic s C e n t e r , University of Minnesota, 250 BioScience Center, 1445 Gortner Avenue, St. Paul, MN 551081095, USA All nematodes used in the test originated from the same delivery of the supplier and the same stock. They were not used in another test. Medium M9, according to G r a t ze r & A hlf (2001) Feeding occurred parallel to test start in the test vessels. Liquid culture of Escherichia coli OP 50, cultured in the test facility in LB-Medium and resuspended in M9 - Medium. (Origin: C a en o r h a b d itis G e n e t ic s C e n t e r , address see above) Company Sanitized. Does not contain T S C A CBf /*'"**% DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE eport Effects on the D evelopm ent of Caenorhabditis elegans in a W ater-S edim en t System acc. to T r a u n s p u r g e r et al. (1997) Page 10 of 20 MProject-No Study-No. J TES T ITEM Pretreatment Test concentration Vehicle Solvent Application technique CONTROL SOLVENT CONTROL , LIQUID CONTROL REFERENCE ITEM The test item was heated to 50 C,'weighed out, dissolved in tert.methyl-butyl-ether and mixed with 70 g quartz sand. The solvent was volatilized out of the quartz sand and the sand mixed with artificial sediment to produce the desired limit test concentration. A limit concentration of 1000 mg /kg artificial sediment was used for the study. 70 g quartz sand ferf.-methyl-butyl-ether 2.4996 g of the test item were weighed out and dissolved in 100 mL /e/t-methyl-butyl-ether. 4 mL of the test item stock solution has been mixed with 70 g quartz sand, necessary for an entire amount of 100 g artificial sediment. After volatilization of the solvent out of the quartz sand, the treated quartz sand was carefully mixed with the artificial sediment to insure a homogeneous distribution of the test item. The amount of quartz sand added was the same for the limit test concentration and the untreated artificial sediment control. After 24 h of equilibration under test conditions 0.5 g of the treated sediment was added to the test vessels. Three replicates with 10 nematodes per replicate using M9-Medium and artificial sediment (without test item) were tested under the same test conditions as the test item replicates. Two replicates with artificial sediment and quartz sand treated with ferf.methyl-butyl-ether without test item were used as solvent control medium. The liquid control solely served as the validity control. M9-Medium (0.5 mL) without sediment was tested using 2 replicates with 10 nematodes per replicate under the same test conditions as the test item replicates. No reference item is recommended according to the literature. TEST METHOD Test duration A static test was performed. 72 h after transfer of the test organisms into the test vessels. Company Sanitized. Does not contain T SCA CBI DR.U.NOACK-IABORATORIUM FR ANGEWANDTE BIOLOGIE Report tects on the Developm ent of Caenorhabditis elegans in a W ater-S edim ent System acc. to T r a u n sp u r g er et al. (1 9 9 7 ) Page 11 of 20 Project-N Study-No. Test vessel Nunc Polystyrol Multiwells (12 wells microtiter plates) Sediment Artificial soil modified according to G r a tzer & A h lf (2 0 0 1 ): - 5 % peat, < 1 mm, air dried and finely ground - 0.5 % dolomite - 70 % quartz sand (with > 50 [%] particles sized between 50 and 200 (jm ) - 4.5 % iron(lil) oxide - 1 .5 % kaolin - 18.5 % aluminium(lll) oxide The pH was determined to be 5.91. Test sediment amount 0.5 g (dry weight, with and without test item) per test well, mixed with 0.5 mL of the food. Food M9-Medium (see above) inoculated with Escherichia coll OP50 (densitiy of approx. 1010 cells/mL) Number of test nematodes per replicate 10 Replicates 5 replicates per limit test concentration, 3 replicates per untreated sediment control, 2 replicates per liquid and solvent control. Age of nematodes First stage juveniles (body length ca. 270 pm) obtained by filtering a fresh culture of nematodes through 10 pm and 5 pm mesh size nylon nets. Preparation of test vessels Prior to test start, 0.5 g sediment mixed with the test item was added into each test vessel. The untreated sediment control vessels were filled with 0.5 g sediment without test item. Introduction of the larvae Prior to test start 0.5 mL of the food medipm was added to the vessels . with the test item treated and the control sediment. 0.5 mL of the food medium was added to the liquid control. Ten selected first stage juveniles were randomly transferred to each test vessel by a sterilized micropipette. After addition of the nematodes, the test vessels were kept on shaking desks and incubated for 72 h. Temperature 20 1 C Light intensity Continuous dark Feeding Food was provided at test start in M9-Medium. posn>fr TSC ftCSl DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE ^Report ' Effects on the Developm ent of Caenorhabditis elegans in a W ater-S edim en t System acc. to T r a u n sp u r g er et al. (1997) Page 12 of 20 fiai?Project-N Study-No KIND AND FREQUENCY OF MEASUREMENT AND OBSERVATIONS Observations were made only at the end of the test (72 h). The number of living nematodes in the liquid control was observed under a microscope at test end to ensure the vitality of the test organisms throughout the exposure. The test vessels were heated (50 C, 5 min) to kill the worms and afterwards the nematodes were stained using an aqueous solution of Rose-Bengal and stored at -20 C until further extraction and measurements. Extraction of the test organisms from sediment The sediment was mixed with approx. 6 mL of diluted Ludox and centrifuged using a swing-out centrifuge (10 min, 700-800 g). The resulting supernatant with the nematodes was decanted in a petri-dish. Criteria of effects The extracted nematodes were observed using a microscope. Body length was measured with a microscale to an accuracy of a t least 0.001 mm. Number of eggs inside the nematodes was counted and recorded. Equipment Laboratory incubator, B 6060 (H eraeus) Microflow biological safety cabinet (MDH) W ater bath, W 350 T (M em m ert) Microscope, LAB 16, (Carl Z eiss) Centrifuge, ZK 364 (Hermle G mbH & Co) Micropipette, Finnpipette Digital (Labsystems OY) Shaking desk, Vortex Genie 2 (Scientific Industries) COURSE OF THE STUDY A preliminary range finding test (NON-GLP) was carried out to determine the concentration range for the main study. Based on the results of this preliminary test, the definitive study was performed at the limit concentration of 1000 mg/kg. _ A sediment control, a solvent control and a liquid control (each without test item) were tested under the same conditions as the test item * treatment. At test end (after 72 h) the worms were examined for survival (liquid control), body length and egg production. Measurements and observations were carried out as described above. tS C A C B i DR.U.NOACK-LABORATORiUM FR ANGEWANDTE BIOLOGIE Report Page 13 of 20 "Effects on the D evelopm ent of Caenorhabditis elegans ** in a W ater-S edim en t System acc. to T r a u n s p u r g e r et al. (1 997) f m !Project-No Study-No. EVALUATION Sediment and solvent control were pooled for statistical analysis since no statistically significant differences were found between these groups using ANOVA. The following evaluations were made: Growth: Body length in pm Egg production: Number of eggs inside the body Fertility: [%] gravid worms (worms with > 1 egg) The number off eggs per worm was calculated for each replicate according to formula 1: R = njnn (1) R = Reproduction ne = Number of eggs at test end n,, = Number of nematodes at test end Fertility [%] of worms was calculated according to formula 2: F = rieX 1 0 0 / nt F = Fertility in % ne = Sum of worms with > 1 egg nt = Sum of worms Introduced per treatment (2) STATISTICAL ANALYSIS ANOVA has been used for comparison of mean values of the replicates. Tests for a normality and homogeneity of variances were conducted prior to ANOVA. P-values for both normality and homogeneity of variances were 0.05. Kru sk al-W allis A N O V A on Ranks was used in case a non-normal data distribution. The a-value for ANOVA and Kr usk a l-W ALUS A NO VA on Ranks was a=0.05. The data for the tables in the report were computer generated and have been rounded for presentation from the full derived data. Consequently if calculated manually based on the given data minor variations may occur from these figures. SOFTWARE Calculations were conducted using the following software: - Sigm aStat rel. 2.0 3 (1 9 9 2 -1 9 9 7 ), S p ss In c . - Microsoft Excel rel. 2 0 0 0 (2000), MICROSOFT CORPORATION C o tn p a W oat conia t s c a g s i DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Effects on the Developm ent of Caenorhabditis elegans in a W a ter-S edim en t System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) Page 14 of 20 m mProject-No, Study-No. VALIDITY CRITERIA Sediment control: - percentage of gravid worms > 80 %. DEVIATIONS FROM THE GUIDELINE DEVIATIONS FROM THE STUDY PLAN REPORTING ARCHIVING None None T h e final report is issued as p aper copy (5 originals) and an electronic record (1 original). The following was retained in the archive of the test facility for the period as specified in the operative national GLP regulations: all raw data study plan (Original 1 of 2) final report paper copy (Original 1 of 5) all records performed by the quality assurance programme including master schedules sample of test and reference items. Microfilms were retained in a safe-deposit by Volksbank, D-31157 Sarstedt. QOtGP&W TSCACB DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report Effects on the Developm ent of Caenorhabditis elegahs w in a W a ter-S edim en t System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) flairPage 15 of 20 Project-N Study-No 4 Results 4.1 Preliminary Test Table 2: Growth and Reproduction of Caenorhabditis etegans (72 h) Test Item Test Object Exposure ^ U Caenorhabditis elegans Artificial sediment L Liquid Control Sediment Control Survival of nematodes 100 Bodylength [pm] SD - Number of eggs per nematode SD - - 1001.4 9 8 .4 4.30 3.61 Gravid worms [%] - 100 [mg product/kg] 10 100 1000 SD = Standard deviation = Not determined - 966.4 74.5 3.27 1 .6 4 - 984.8 67.1 2.60 1 .5 7 - 962.4 67.8 3.60 1.61 100 100 100 noA' CO' pos c&v DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE I Report Effects on the Developm ent of Caenorhabditis elegans mm in a W ater-S edim en t System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) Page 16 of 20 B0Project-N Study-No 4.2 Definitive Test Measurement of bodylength after 72 hours is given in Tab. 3. Data of egg production after 72 hours are presented in Tab. 4. Effect values are given in Tab. 5. Table 3: Bodylength [pm] after 72 hours Concentration fmq/kg] Rep. 1 Rep. 2 1000 Rep. 3 Rep. 4 Rep. 5 Solvent Rep. 1 Control Rep. 2 Rep. 1 Control Rep. 2 Rep. 3 1 672 1200 672 1080 1032 528 1224 1176 1296 936 2 I3 936 1152 696 960 720 1440 1224 1152 696 552 1240 840 576 672 984 1200 768 1128 1248 600 4 720 624 1152 1104 1182 1152 912 816 696 840 Nematode 56 1200 648 1032 672 768 936 576 888 1248 936 1176 672 1008 936 1512 912 1080 504 912 864 7 696 864 1080 936 696 840 1032 984 1008 864 8 912 768 1200 864 888 648 912 1008 1200 1008 9 1104 1128 672 888 579 960 840 1152 984 960 10 696 672 864 696 888 1176 1056 936 936 Table 4: Num ber o f eggs after 72 hours Concentration Nematode [mg/l 1 2 3 4 5 6 7 8 9 10 Rep. 1 1 2 2 1 8 3 1 2 4 1 Rep. 2 7 1 2 0 3 2 2. 1 3 1 1000 Rep. 3 1 1 11 14 2 3 2 7 1 3 Rep. 4 2 4 3 0 0 12 1 2 - Rep. 5 3 2 0 5 9 2 2 3 11 2 Solvent Rep. 1 0 12 1 13 21 3 2 1 Control Rep. 2 27 1 1 1 0 2 19 2 12 1 10 Rep. 1 1 3 0 7 11 1 2 32 2 Control Rep. 2 11 0 7 1 6 1 2 13 2 2 Rep. 3 3 11 I 1 1 2 2 0 3 1 2 - : Nematode could not be found after 72 h DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report Effects on the Developm ent of Caenorhabditis elegans in a W ater-S ed im en t System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) Page 17 of 20 Project-N Study-No Table 5: Growth, Number-of eggs and Fertility after 72 hours (SD = Standard Deviation) Treatment Replicate Bodylength [pm] Clontrol 123 1080 960 917 Mean value SD 985.7 84.5 Solvent Ccintrol 1 2 Mean value SD 928 895 911.5 23.3 Eggs per Nematode 3.2 4.5 2.6 3.43 3.67 5.6 6.4 6.00 8.13 Gravid Nematodes* [%] 90 90 90 90.0 0.0 90 90 90.0 0.0 Treatment Replicate 12 1000 mg/kg 3 4 5 Mean value Sig. SD Dif.** Bodylength [pm] ' 874 862 929 992 850 901.4 59.0 No Eggs per Nematode 2.5 2.2 4.5 1.7 3.9 2.98 3.05 No Gravid Nematodes* 100 90 100 78 [%] * Percentage of Nematodes with > 1 egg ** significantly different from the pooled control (ANOVA, a < 0.05) - not tested 90 92.0 9.10 - o op o p* DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report Effects on the Developm ent of Caenorhabditis elegans in a W ater-S ed im en t System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) Page 18 o f 20 Project-N Study-No. 5 Validity Criteria The study was performed according to T r au n spur g er et al. (1997) and the principles of GLP. The results met the validity criteria: Percentage of gravid worms in the sediment control > 80 % (in this study 90 %). 6 Conclusions After 72 hours the NOEC o i determined to be greater than 1000 mg/kg based on nominal test item concentrations. rwas 7 Literature / References G r a t z e r , H. & W . A hlf (2001): Adjustment of a formulated sediment for sediment testing with Caenorhabditis elegans (Nematoda) Acta hydrochim. Hydrobiol. 29:41-46. - HOss, S., Ha ij z e r , M., T r a u n s p u r g e r , W . & C .E.W . S t e in b e r g (1999): Growth and Fertility of Caenorhabditis elegans (Nematoda) in unpolluted freshwater sediments: Response to particle size distribution and organic content. Environ. Tox. and Chem. 18:2921-2925. - T r a u n s p u r g e r , W ., Ha it z e r , M , HOss , S., B e ie r , S., A h l f , W . & C. St e in b e r g (1997): Ecotoxicological assessment of aquatic sediments with Caenorhabditis elegans (Nematoda) - A method for testing liquid medium and whole-sediment samples. Environ. Tox. and Chem. 16:245250. 0 60s G o' DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Effects on the D evelopm ent of Caenorhabditis elegans in a W ater-S edim ent System acc. to T r a u n s p u r g e r et al. (1 9 9 7 ) Page 19 of 20 lo tProject-N Study-No. 8 Certificate of Test Item Analysis G iriu u GmbH. W e it Gendoif CQG / QuatsmenagemeM W 450A Burgfcirdtea F u : +49 8679 / 2676 Clariant Inspection certificate a c c o rd in g to EN10204-3.1B D ate: 13.11.2001 Page: 1 / 1 Our consignm ent M aterial M ate rial-r.o . Baton No. Cti m e bate:*./ o r w n ic r 't h e co n sig n m en t i s a p a r t , th e f o llo w in g v a lu e s were detarmir.ee. They contoxm to vii* agreed product specification. In sp ectio n ch a ra cter actit/ '-fn e th o d S p e cifica tio n R esult The above p a r t ic u la r s do not r e le a s e th e custom er from th e o b lig a t io n c a r r y o u t ar. inspection or goods received. to Ms. Haumg-srrner - Wcr<sinspector Gom pm tf Snrtized. Does not contain TSC A C BI DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report Page 20 of 20 Effects on the Developm ent of Caenorhabditis elegans 00 in a W ater-S edim en t System acc. to T ra u n sp u r g er et al. (1 9 9 7 ) SlProject-N Study-No. 9 Certificate of Dr.U.Noack-Laboratorium n N iederschsisches U m w eltm inisterium Gute Laborpraxis/Good Laboratory Practice GLP-Bescheinigung/Statement o f GLP Compliance (gema/according to 1 9 b Abs.1 Chemikaliengesetz) Eine GLP-Inspektlon zur berwachung der Einhaltung der GLP-Grundstze gem Chemikaliengesetz bzw. Richtlinie 88/320/EG wurde durchgefhrt ln: Assessment of conformity with GLP according to Chemikaliengesetz and Directive 88/320/EEC a t X Prfeinrichtung / Test facility Prfstandort /T e s t site D r. U . N O A C K LA B O R A TO RIUM fQr angew andte Biologie K the-P aulus-S tr. 1 D -3 1157 Sarstedt D r. U. NOACK LABO RATO RY o f applied biology K the-Paulus-Str. 1 D -3 1 157 Sarstedt (Unverwechselbare Bezeichnung und Adresse/Unequlvocal name snd address) Prfungen nach Kategorien/Areas of Expertise (flsmift/Seconfing ChetriVwV-GLP Nr. SJ/OECD guidance) 1 - Prfungen zu r Bestim m ung der physikalisch-chem ischen Eigenschaften und Gehaltsbestim m ungen 3 - Prfungen zu r Bestim m ung der erbgutverndem den Eigenschaften (in vitro und in vivo) 4 - Um w eittoxikologische Prfungen zu Auswirkungen au f aquatische und terrestrische O rganism en 5 - Prfungen zum V erhalten im Boden, im W asser und in der Luft, Prfungen zu r Bioakkum ulation und zur M etabollsierung 6 - Prfungen zu r Bestim m ung von Rckstnden 7 - Prfungen zu r Bestim m ung der Auswirkungen auf Mesokosm en und natrliche kosystem e Datum d e r Inspektion / D ate o f Inspectlon (Tas.MonaUahr / day-month.year) 2 5 .-2 7 . Juni 2001 / June 25 * - 27* 2001 29. August und 21. September 2001 / August 29 * and September 2 1 *, 2001 Die genannte PrOfelnrichtung/Por genannte Prttfstandoit befindet sich Im nationalen GLP- Oberwachungsverfahren und wird regelmig auf Einhaltung dor GLP-Grundstze berwach!. Auf der Grundlage des Inspekdonabedchtes wird hlenntt besttigt, dass kt dieser PrtMelnricNung/dleeem Prftfetandort die oben genannten Prfungen unter Einhaltung der GLP-Grundstze durchgefOhit werden knnen. Niederschsisches Umweltministerium Referat 33 Archivstrae 2 30169 Hannover The above mentioned teat facility / test cilo is Included in the national GLP Compliance Programme and Is Inspected on a regular basis. Based on the inspection report It can be confirmed, that this test facility I toot..silo Is able to conduct the aforementioned studies In cemplianca with the Principles of GLP. 11. Marz 2002 Im Auftraga s Dr. Braedt :^6 G & & <6*