Document N2j79p6R80BOvOyLXQvXVQR7b

AR226-1431 DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- Effects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Sponsor Telomer Research Project ASAHI GLASS CLARIANT DAIKIN INDUSTRIES LTD DUPONT RECEIVED OPPT NCIC 2003 July 18 1:09PM Author Gunda Winkelmann Test Facility DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Kthe-Paulus-Strae 1 D-31157 Sarstedt Laboratory Project ID Project-No. 020719CK Study-No. RNA84111 Study-No.(Study plan) RNA8411- Study Completed On 2003-04-14 Report Page 1 of 20 DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 2 of 20 Project-No. 020719CK Study-No. RNA84111 Statement of GLP Compliance Title 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- Effects on the Development of Caenorhabditis elegans in a Water- Sediment System Guideline TRAUNSPURGER et al. (1997) Test Item 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro(Batch number: P00/001) Test Facility DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Kthe-Paulus-Str.1, D-31157 Sarstedt Phone: (+49) 50 66 / 706 70, Fax: (+49) 50 66 / 706 789 E-mail: info@noack-lab.de Deviations from GLP Principles None We 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) DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 3 of 20 Project-No. 020719CK Study-No. RNA84111 Statement of the Quality Assurance Unit Title Guideline Test Item 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a WaterSediment System TRAUNSPURGER et al. (1997) 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro(Batch number: P00/001) Study Director Gunda Winkelmann The 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. The accordance of the study with its study plan and the principles of Good Laboratory Practice is guaranteed. 2003-04-14 ...pp..................................................... Gudrun Mhrmann-Kalabokidis DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 4 of 20 Project-No. 020719CK Study-No. RNA84111 Personnel Involved Study Director: Deputy: Technical Staff: Quality Assurance Unit: Gunda Winkelmann (Engineer of Horticulture) Dirk Scheerbaum (Biologist) Karin Ruthenberg Gudrun Mhrmann-Kalabokidis (Biologist) Deputy: Susanne Becker (Biologist) Director of the Testing Facility: Dr. Udo Noack (Biologist) DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 5 of 20 Project-No. 020719CK Study-No. RNA84111 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 DR.U.NOACK-LABORATORIUM Page 1 2 3 4 5 6 7 8 9 15 15 16 18 18 18 19 20 DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 6 of 20 Project-No. 020719CK Study-No. RNA84111 List of 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 [m] after 72 hours Number of eggs after 72 hours Growth, Number of eggs and Fertility after 72 hours Page 7 15 16 16 17 DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 7 of 20 Project-No. 020719CK Study-No. RNA84111 1 Summary The effects of the test item 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- (batch no. P00/001) on the development of the free living soil nematode Caenorhabditis elegans were determined in a water-sediment system. The study was carried out according to TRAUNSPURGER et al. (1997). Test duration was 72 hours. The study was conducted from 2002-11-22 to 2002-11-25 at DR.U.NOACKLABORATORIUM FR ANGEWANDTE BIOLOGIE, 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 (tert.-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 item 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- are summarised in Table 1. Table 1: Growth and Reproduction of Caenorhabditis elegans (72 h) Test Item Test Object 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroCaenorhabditis elegans Exposure Artificial sediment Liquid Control Sediment Control Solvent Control Survival of nematodes [%] 100 - - Bodylength [m] SD - 985.7 84.5 911.5 23.3 Number of eggs per nematode SD - 3.43 3.67 6.00 8.13 Gravid worms* [%] - 90.0 0.0 90.0 0.0 [mg product/kg] 1000 - 901.4 59.0** SD = Standard deviation * = Percentage of nematodes with > 1 egg ** = No significant difference compared to the pooled control - = Not determined 2.98 3.05** 92.0 9.10 DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 8 of 20 Project-No. 020719CK Study-No. RNA84111 2 Characterisation Data of the Test Item TEST ITEM Batch Number CAS RN Chemical characterization Purity Appearance Water solubility Stability in water Density 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- P00/001 678-39-7 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluorodecane-1-ol (EINECS) C8-perfluoroalkylethanol: 99.8 % (a/a), sum perfluoroalkylethanol 99.9 % (a/a) Yellowish solid wax 134 g/L at 12 degree C, Std 29 g/L, n=8 (DuPont, 2002) 137 g/L at 25 degree C, Std 53 g/L, n=17 (DuPont, 2002) Stable at test conditions Ca. 1.7 g/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. DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 9 of 20 Project-No. 020719CK Study-No. RNA84111 3 Method TEST GUIDELINE TYPE AND PURPOSE OF THE STUDY TEST SYSTEM Reason for the selection Source Food medium Feeding Food TRAUNSPURGER et al. (1997): "Ecotoxicological assessment of aquatic sediments with Caenorhabditis elegans (nematoda) - A method for testing in liquid medium and whole sediment samples." Determination of the potential impact of 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- on the free living soil nematode Caenorhabditis elegans. Caenorhabditis elegans (nematoda) N2, var. Bristol, wild type lab strain According to the literature (TRAUNSPURGER 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: CAENORHABDITIS GENETICS CENTER, 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 GRATZER & AHLF (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: CAENORHABDITIS GENETICS CENTER, address see above) DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 10 of 20 Project-No. 020719CK Study-No. RNA84111 TEST ITEM Pretreatment Test concentration Vehicle Solvent Application technique CONTROL SOLVENT CONTROL LIQUID CONTROL REFERENCE ITEM TEST METHOD Test duration 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- 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 tert.-methyl-butyl-ether 2.4996 g of the test item were weighed out and dissolved in 100 mL tert.-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 tert.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. A static test was performed. 72 h after transfer of the test organisms into the test vessels. DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 11 of 20 Project-No. 020719CK Study-No. RNA84111 Test vessel NUNC Polystyrol Multiwells (12 wells microtiter plates) Sediment Artificial soil modified according to GRATZER & AHLF (2001): - 5 % peat, < 1 mm, air dried and finely ground - 0.5 % dolomite - 70 % quartz sand (with > 50 [%] particles sized between 50 and 200 m) - 4.5 % iron(III) oxide - 1.5 % kaolin - 18.5 % aluminium(III) 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 coli OP50 (densitiy of approx. 1010 cells/mL) Number of test nematodes 10 per replicate 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 m) obtained by filtering a fresh culture of nematodes through 10 m and 5 m 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 medium 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. DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 12 of 20 Project-No. 020719CK Study-No. RNA84111 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 at least 0.001 mm. Number of eggs inside the nematodes was counted and recorded. Equipment Laboratory incubator, B 6060 (HERAEUS) Microflow biological safety cabinet (MDH) Water bath, W 350 T (MEMMERT) Microscope, LAB 16, (CARL ZEISS) Centrifuge, ZK 364 (HERMLE GMBH & 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. DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 13 of 20 Project-No. 020719CK Study-No. RNA84111 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 m - 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 = ne/nn (1) R = Reproduction ne = Number of eggs at test end nn = Number of nematodes at test end Fertility [%] of worms was calculated according to formula 2: F = nex 100 / nt (2) F = Fertility in % ne = Sum of worms with > 1 egg nt = Sum of worms introduced per treatment STATISTICAL ANALYSIS SOFTWARE 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. KRUSKAL-WALLIS ANOVA on Ranks was used in case a non-normal data distribution. The a-value for ANOVA and KRUSKAL-WALLIS ANOVA 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. Calculations were conducted using the following software: - SigmaStat rel. 2.03 (1992-1997), SPSS INC. - Microsoft Excel rel. 2000 (2000), MICROSOFT CORPORATION DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 14 of 20 Project-No. 020719CK Study-No. RNA84111 VALIDITY CRITERIA Sediment control: - percentage of gravid worms > 80 %. DEVIATIONS FROM THE GUIDELINE DEVIATIONS FROM THE STUDY PLAN REPORTING ARCHIVING None None The final report is issued as paper 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. DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 15 of 20 Project-No. 020719CK Study-No. RNA84111 4 Results 4.1 Preliminary Test Table 2: Growth and Reproduction of Caenorhabditis elegans (72 h) Test Item Test Object Exposure Liquid Control Sediment Control 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroCaenorhabditis elegans Artificial sediment Survival of nematodes 100 - Bodylength [m] SD - 1001.4 98.4 Number of eggs per nematode SD - 4.30 3.61 Gravid worms [%] - 100 [mg product/kg] 10 100 1000 - 968.4 74.5 3.27 1.64 100 - 984.8 67.1 2.60 1.57 100 - 962.4 67.8 3.60 1.61 100 SD = Standard deviation - = Not determined DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 16 of 20 Project-No. 020719CK Study-No. RNA84111 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 [m] after 72 hours Concentration [mg/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 2 936 696 720 1440 696 1240 576 984 768 1248 3 1152 960 1224 1152 552 840 672 1200 1128 600 4 720 624 1152 1104 1182 1152 912 816 696 840 Nematode 5 6 1200 648 1032 672 768 936 576 888 1248 936 1176 1008 672 936 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: Number of eggs after 72 hours Concentration Nematode [mg/kg] 1 2 3 4 5 6 Rep. 1 1 2 2 1 8 3 Rep. 2 7 1 2 0 3 2 1000 Rep. 3 1 1 11 14 2 3 Rep. 4 2 4 3 0 0 1 Rep. 5 3 2 0 5 9 2 Solvent Rep. 1 0 12 1 13 21 3 Control Rep. 2 27 1 1 1 0 2 Rep. 1 1 3 0 7 11 1 Control Rep. 2 11 0 7 1 6 1 Rep. 3 3 11 1 1 2 2 - : Nematode could not be found after 72 h 7 8 9 10 1 2 4 1 2 1 3 1 2 7 1 3 2 1 2 - 2 3 11 2 2 1 1 2 19 2 1 10 2 3 2 2 2 13 2 2 0 3 1 2 DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 17 of 20 Project-No. 020719CK Study-No. RNA84111 Table 5: Growth, Number of eggs and Fertility after 72 hours (SD = Standard Deviation) Treatment Replicate Bodylength [m] Control 1 2 3 1080 960 917 Mean value SD 985.7 84.5 Solvent Control 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 1000 mg/kg Replicate 1 2 3 4 5 Bodylength [m] 874 862 929 992 850 Eggs per Nematode 2.5 2.2 4.5 1.7 3.9 Gravid Nematodes* 100 90 100 78 90 [%] * Percentage of Nematodes with > 1 egg ** significantly different from the pooled control (ANOVA, a < 0.05) - not tested Mean value Sig. SD Dif.** 901.4 No 59.0 2.98 3.05 No 92.0 9.10 - DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 18 of 20 Project-No. 020719CK Study-No. RNA84111 5 Validity Criteria The study was performed according to TRAUNSPURGER 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 of 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- was determined to be greater than 1000 mg/kg based on nominal test item concentrations. 7 Literature / References - GRATZER, H. & W. AHLF (2001): Adjustment of a formulated sediment for sediment testing with Caenorhabditis elegans (Nematoda) Acta hydrochim. Hydrobiol. 29:41-46. - HSS, S., HAITZER, M., TRAUNSPURGER, W. & C.E.W. STEINBERG (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. - TRAUNSPURGER, W., HAITZER, M, HSS, S., BEIER, S., AHLF, W. & C. STEINBERG (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. DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 19 of 20 Project-No. 020719CK Study-No. RNA84111 8 Certificate of Test Item Analysis DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoroEffects on the Development of Caenorhabditis elegans in a Water-Sediment System acc. to TRAUNSPURGER et al. (1997) Page 20 of 20 Project-No. 020719CK Study-No. RNA84111 9 Certificate of DR.U.NOACK-LABORATORIUM