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AR226-1433 DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoro- Terrestrial Plants Toxicity, Seedling Emergence according to OECD Guideline 208 (Draft July 2000) Sponsor Telomer Research Project Asahi Glass Cl a r ia n t Da ik in In d u s t r ie s Lt d Du p o n t RECEIVED OPPT NCIC 2003 July 18 1:09PM Author Silke Fiebig Test Facility Dr .U.No a c k -La b o r a t o r iu m FR ANGEWANDTE BIOLOGIE Kthe-Paulus-Str. 1 D-31157 Sarstedt Laboratory Project ID Project-No. 020719CK Study-No. TNC84113 Study No. of the study plan: TNC8411- Page 1 of 51 Date 2003-05-02 DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report Page 2 of 51 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- Terrestrial Plants Toxicity, Seedling Emergence Project-No. 020719CK acc. to OECD Guideline 208 (Draft July 2000)_____________________Study-No. TNC84113 Statement of GLP Compliance Title 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoroTerrestrial Plant Toxicity, Seedling Emergence Guideline OECD 208 (Draft document, July, 2000), Terrestrial (Non Target) Plant Test, (Part A) Seedling Emergence Test Item 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoro- (Batch number: P00/001) Test Facility Dr .U.No a c k -La b o r a t o r iu m f r a n g e w a n d t e Bio l o g ie Kthe-Paulus-Str.1, D-31157 Sarstedt Phone: (+49) 050 66 / 706 70, Fax: (+49) 050 66 / 706 789 E-mail: info@noack-lab.de Deviations of GLP Regulations None We declare that this study was conducted and reported in compliance with the actual OECD princi ples of Good Laboratory Practice and the national GLP regulations as specified in the law in force, except deviations mentioned above. 2003-05-02 (Date) (Silke Fiebig, Study Director) 2003-05-02 (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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 3 of 51 Project-No. 020719CK Study-No. TNC84113 Statement of the Quality Assurance Unit Title Guideline Test Item Study Director 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoroTerrestrial Plant Toxicity, Seedling Emergence OECD 208 (Draft document, July, 2000), Terrestrial (NonTaget) Plant Test, (Part A) Seedling Emergence 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoro- (Batch number: P00/001) Silke Fiebig The study was verified as follows: inspection study plan study based report dates 2002-11-13 2002- 1 1 -2 1 2003-02-19 2003-02-20 2003-02-21 2003-04-09 2003-04-11 2003-04-29 date of report 2002-11-13 2002- 1 1 -2 1 2003-02-20 2003-02-20 2003-02-21 2003-04-10 2003-04-11 2003-04-29 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. 2003-05-02 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 4 of 51 Project-No. 020719CK Study-No. TNC84113 Personnel Involved Study Director: Silke Fiebig (Engineer, Biotechnologist) Technical Staff: Kirsten McVean Petra Makus Simone Pioch Barbara Wienker Quality Assurance Unit: 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 5 of 51 Project-No. 020719CK Study-No. TNC84113 Table of Content Title Page Statement of GLP Compliance Page 1 2 Statement of the Quality Assurance Unit 3 Personnel Involved 4 Table of content List of Tables 5 6 List of Figures 7 1 Summary 2 Characterisation Data of the Test Item 8 10 3 Method 11 4 Results 20 4.1 Visual Phytotoxicity 20 4.2 Shoot Height 22 4.3 Fresh W eight (Biomass) 23 4.4 Emerged Seedlings 25 4.5 Effect Levels 26 4.5.1 Inhibition of Shoot Height, Fresh Weight and Number of Emerged Seedlings 26 4.5.2 EC25, EC50and NOEC Values 27 5 Graphs 28 6 Conclusions 29 7 Validity Criteria 29 8 Literature 30 9 Biological Data 31 9.1 Observation Parameter 31 9.2 Emerged Seedlings on Day 7 32 9.3 Emerged Seedlings on Day 14 33 9.4 Emerged Seedlings on Day 21 35 9.5 Visual Phytotoxicity on Day 7 36 9.6 Visual Phytotoxicity on Day 14 38 9.7 Visual Phytotoxicity on Day 21 39 9.8 Shoot Height 40 9.9 Fresh W eight (Biomass) Day 21 47 10 Certificate of Test Item Analysis 50 11 GLP Certificate of Dr .U.No a c k -La b o r a t o r iu m 51 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 6 of 51 Project-No. 020719CK Study-No. TNC84113 List of Tables Table 1: Shoot Height: NOEC, EC25 and EC50 Values with Confidence Range (p) Table 2: Fresh Weight: NOEC, EC25 and EC50 Values with Confidence Range (p) Table 3: Number of Emerged Seedlings: NOEC, EC25 and EC50 Values with Confidence Range (p) Table 4: Oats: Phytotoxic Effects and Rate of Emergence after 21 Days Table 5: Rape: Phytotoxic Effects and Rate of Emergence after 21 Days Table 6: Soybean: Phytotoxic Effects and Rate of Emergence after 21 Days Table 7: Inhibition of Shoot Height Table 8: Inhibition of Fresh Weight Table 9: Inhibition of Emerged Seedlings Table 10: Oats: Emerged Seedlings on Day 7 Table 11: Rape: Emerged Seedlings on Day 7 Table 12: Soybean: Emerged Seedlings on Day 7 Table 13: Oats: Emerged Seedlings on Day 14 Table 14: Rape: Emerged Seedlings on Day 14 Table 15: Soybean: Emerged Seedlings on Day 14 Table 16: Oats: Emerged Seedlings on Day 21 Table 17: Rape: Emerged Seedlings on Day 21 Table 18: Soybean: Emerged Seedlings on Day 21 Table 19: Oats: Visual Observations and Number of Dead Plants on Day 7 Table 20: Rape: Visual Observations and Number of Dead Plants on Day 7 Table 21: Soybean: Visual Observations and Number of Dead Plants on Day 7 Table 22: Oats: Visual Observations and Number of Dead Plants on Day 14 Table 23: Rape: Visual Observations and Number of Dead Plants on Day day 14 Table 24: Soybean: Visual Observations and Number of Dead Plants on Day 14 Table 25: Oats: Visual Observations and Number of Dead Plants on Day 21 Table 26: Rape: Visual Observations and Number of Dead Plants on Day 21 Table 27: Soybean: Visual Observations and Number of Dead Plants on Day 21 Table 28 : Oats: Shoot Height [cm] Pooled Control, 10 and 32 mg/kg DW Table 29 : Oats: Shoot Height [cm] 100, 320 and 1000 mg/kg DW Table 30 : Rape: Shoot Height [cm] Pooled Control, 6.25 and 12.5 mg/kg DW Table 31 : Rape: Shoot Height [cm] 25, 50 and 100 mg/kg DW Table 32 : Soybean: Shoot Height [cm] Pooled Control, 10 and 32 mg/kg DW Table 33 : Soybean: Shoot Height [cm] 100, 320 and 1000 mg/kg DW Page 9 9 9 20 21 21 26 27 27 32 32 33 33 34 34 35 35 36 36 37 37 38 38 39 39 40 40 41 42 43 44 45 46 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 7 of 51 Project-No. 020719CK Study-No. TNC84113 Table 34: Table 35: Table 36: Oats: Fresh W eight on Day 21 Rape: Fresh W eight on Day 21 Soybean: Fresh W eight on Day 21 List of Figures Figure 1: Oats: Shoot Height after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil Figure 2: Rape: Shoot Height after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil Figure 3: Soybean: Shoot Height after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil Figure 4: Oats: Fresh W eight after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil Figure 5: Rape: Fresh W eight after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil Figure 6: Soybean: Fresh W eight after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil Figure 7: Oats: Emerged Seedlings after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil Figure 8: Rape: Emerged Seedlings after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil Figure 9: Soybean: Emerged Seedlings after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil Figure 10: Rape: Shoot Height after 21 d Figure 11: Rape: Fresh W eight after 21 d 47 48 49 Page 22 22 23 23 24 24 25 25 26 28 28 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 8 of 51 Project-No. 020719CK Study-No. TNC84113 1 Summary The phytotoxicity of the test item 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoro- (batch number P00/001) to three terrestrial plant species was determined in a seedling emergence test over a period of 21 days, from November 2 1,2 00 2 to February 07, 2003 (for detailed test periods see test conditions) according to OECD 208 Draft document, July 2000 at the Dr .U.No a c k -La b o r a t o r iu m f r a n g e w a n d t e Bio l o g ie , Sarstedt, Germany. The test was conducted with the nominal concentrations 10.0 - 32.0 - 100 - 320 - 1000 soil dry weight (DW) (oats and soybean) 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/kg soil dry weight (DW) (rape) selected on the basis of a preliminary phytotoxicity test. To reach a sorption equilibrium the application of the test medium was performed 24 h be fore the seeds were sown. Test systems were one monocotyledon (oats (Gramineae)) and two dicotyledons (rape (Brassicaceae) and soybean (Leguminosae)). The test containers were bottom watered with nutrient solution as needed. A solvent control without test item (quartz sand only spiked with tert.-buthyl-methyl-ether) was tested under the same conditions as the test groups. To exclude effects of the solvent tert.-buthyl-methyl-ether a second control without solvent was tested. The solvent tert.buthyl-m ethyl-ether caused no inhibitory effects in shoot height, biomass growth, and rate of emergence. No statistically significant differences were found between the control and the solvent control. For rape, enhanced growth was observed for the solvent control relative to the control. This promotion was not significant for this study. Therefore for calculations the replicates of the control and solvent control were summarized in a pooled control group. The toxic effects of the test item were determined on day 7, 14 and 21 by visual observa tions (number of emerged seedlings, phytotoxic effects and number of dead plants) and on day 21 by shoot height and shoot fresh weight determination. Shoot height, fresh weight and number of emerged seedlings were checked for statistically significant differences relative to the controls and the NOEC, the EC25- and EC50-values were calculated (Table 1-3). 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 9 of 51 Project-No. 020719CK Study-No. TNC84113 Table 1: Shoot Height: NOEC, EC25 and EC50 Values with Confidence Range (p) S p e c ie s NOEC* EC25 [m g/kg DW] [m g/kg DW] p = 95% EC50 [m g/kg DW] [m g/kg DW] p = 95% [m g/kg DW] oats rape 1000 50.0 > 1000 80.2 65.6 - 97.9 > 1000 > 100 - soybean 1000 > 1000 - > 1000 - Table 2: Fresh Weight: NOEC, EC25 and EC50 Values with Confidence Range (p) S p e c ie s NOEC* EC25 [m g/kg DW] [m g/kg DW] p = 95% EC50 [m g/kg DW] [m g/kg DW] p = 95% [m g/kg DW] oats 1000 > 1000 - > 1000 - rape 25.0 51.7 44.0 - 60.8 73.7 62.6 - 86.7 soybean 320 > 1000 - > 1000 - Table 3: Number of Emerged Seedlings: NOEC, EC25 and EC50 Values with Confidence Range (p) S p e c ie s NOEC* EC25 [mg/kg DW] [m g/kg DW] p = 95% EC50 [m g/kg DW] [m g/kg DW] p = 95% [m g/kg DW] oats 1000 > 1000 - > 1000 - rape 100 > 100 - > 100 - soybean 1000 > 1000 - > 1000 - - not determinable * The highest tested concentration of the test item at which no statistically significant effect is observed relative to control 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 10 of 51 Project-No. 020719CK Study-No. TNC84113 2 Characterisation Data of the Test Item TEST ITEM 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoro- Batch Number P00/001 CAS RN 678-39-7 Chemical characterization 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluorodecane-1ol (EINECS) Purity C8-perfluoroalkylethanol: 99.8 % (a/a), sum perfluoroalkylethanol 99.9 % (a/a) Appearance Yellowish solid wax Water solubility 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) Stability in water Stable at test conditions Density Ca. 1.7 g/cm 3 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 11 of 51 Project-No. 020719CK Study-No. TNC84113 3 Method GUIDELINE OECD 208 (Draft document, July 2000), Terrestrial (Non Target) Plants Testing, (Part A) Seedling Emergence TYPE AND PURPOSE OF THE STUDY Principle of the test TEST SYSTEMS Reason for the selection of the test systems Origin Storage Determination of toxic effects of soil incorporated chemicals on the emergence of seedlings and the early stages of growth of three terrestrial plant species. The test item was incorporated into the soil in which the seeds were sown. During the test period the number of seedlings emerged were counted every 7 days. The seedlings also were observed for visual phytotoxic symptoms. A fter 21 days the plants were harvested and shoot height and shoot fresh weight were measured. Three plant species of different families were tested: Monocotyledonae: Avena sativa (oats)1, Flamingsstern Gramineae Dicotyledonae: Glycine max (soybean) , Jutro Brassica napus (rape) , Pollen Leguminosae Brassicaceae The species are recommended species according to the test guideline. 1 Lo c h o w Pe t k u s G m b H, Postfach 1197, D-29296 Bergen 2 Su d w e s t s a a t Gbr., Im Rheinfeld 1-13, D-76437 Rastatt 3 G r o t z n e r Pf l a n z e n z u c h t G m b H & Co . KG, M argaretenhof 23, D-22397 Hamburg The seeds were stored at the test facility at room temperature (20 5 C) and protected from light and moisture until use. 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 12 of 51 Project-No. 020719CK Study-No. TNC84113 TEST ITEM 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heptadecafluoro- Pretreatment The test item was heated to 50 C, weighed out, dissolved in fe/f.-m ethyl-buthyl-ether and mixed with quartz sand. After the solvent had volatilised out of the quartz sand, the quartz sand was mixed with test soil to provide the test treatments. Test concentration Oats, soybean: 1000 - 320 - 100 - 32 - 10 mg/kg soil dry weight (DW) (factor V10) Rape: 100 - 50 - 25 - 12.5 - 6.25 mg/kg soil dry weight (DW) (factor 2) Vehicle Max. 1 % Quartz sand Solvent fe /f.-m e thyl-b uth yl-e the r Stock solutions for oats study The soil test concentrations of 10 - 100 mg/kg DW were made using two freshly prepared stock solutions dissolved in fe/f.-m ethyl-butyl-ether having test concentrations of 25 and 10 g/L. Appropriate stock solution volumes were mixed with quartz sand and finally with the soil to give specified soil con centrations (see table below). The test concentrations 320 and 1000 mg/kg DW were prepared by adding directly weighed quantities of the test item to measured quantity of solvent (fe/f.-methyl-butyl-ether) (see table below). The volume of stock solution mixed with quartz sand is given below. Soil te st concentration [m g/kg DW] Amount of DW soil [kg] Amount of Conc. of quartz Test Item sand S to c k (1 %) solution [g] [g/L] 1000 10.5 105.2 - 320 10.5 105.2 - S to c k solution volum e* [m L ] - D irect weighing nominal actual [g] 10.521 3.367 [g] 10.5213 3.3671 Stock solution volum e prepared [m L ] - - S o lv e n t [m L ] 35 20 100 1 5 .8 1) 157.8 25 63.1 2.5 2.4996 100 - 32 10.5 105.2 25 13.6 - - -- 10 10.5 105.2 10 11.0 0.5 0.5003 50 - * Volume of stock solution (ferf.-methyl-butyl-ether) mixed with quartz sand DW = dry weight Conc. = concentration 1) = larger amount of DW soil prepared because rape tested additionally 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 13 of 51 Project-No. 020719CK Study-No. TNC84113 Stock solutions for soybean study The soil test concentrations of 10 - 100 mg/kg DW were made using two freshly prepared stock solutions dissolved in fe/f.-m ethyl-butyl-ether having test concentrations of 50 and 5 g/L. Appropriate stock solution volumes for the concentrations 32 and 100 mg/kg DW were mixed with quartz sand and finally with the soil to give specified soil concentrations (see table below). The test concentrations 320 and 1000 mg/kg DW were prepared by adding directly weighed quantities of the test item to measured quantity of solvent (fe/f.-m ethyl-butyl-ether) (see table below). The stock solution for the concentration 10 mg/kg DW was prepared by 1:10 dilution of the stock solution 50 g/L (see ta ble below). The volume of stock solution mixed with quartz sand is given below. Soil te st concentration [m g/kg DW] Amount of DW soil [kg] Amount of Conc. of quartz Test Item sand S to c k (1 %) solution [g] [g/L] 1000 5.2 51.9 - 320 5.2 51.9 - 100 5.2 51.9 50 32 5.2 51.9 50 10 5.2 51.9 5 S to c k solution volum e* [m L ] - D irect weighing nominal actual S to c k solution volum e prepared [g] [g] [mL] 5.186 5.1863 - S o lv e n t [m L ] 30 - 1.659 1.6591 - 25 10.4 2.5 2.4998 50 - 3.4 - - - - 10.0 -- - - * Volume of stock solution (ferf.-methyl-butyl-ether) mixed with quartz sand DW = dry weight Conc. = concentration Stock solutions for rape study A stock solution for 1-Decanol,3,3,4,4,5,5,6,6,7,7,8,8,9,9, 10 ,10 ,10-heptadecafluoro- for soil concentrations 10 - 1000 mg/kg DW (57.8 g/L; nominal: 2.89 g, actual: 2.8901 g dissolved in 50 mL of ferf.-methyl-butyl-ether) was freshly prepared. The soil test concentrations 6.25 - 50 mg/kg DW were pre pared by 1:2 dilution of the stock solution with a final volume of 50 mL each. 10 mL of the appropriate stock solution was mixed with quartz sand. 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 14 of 51 Project-No. 020719CK Study-No. TNC84113 Soil test concen Amount of tration DW soil [mg/kg DW] 100 50 25 12.5 [kg] 5.8 5.8 5.8 5.8 Amount of Stock solu quartz sand tion volum e* (1 %) [g] 57.8 57.8 57.8 57.8 [mL] 10 10 10 10 Conc. of Test Item Stock solu tion [g/L] 57.8 28.9 14.5 7.2 6.25 5.8 57.8 10 3.6 * Volume of stock solution (ferf.-methyl-butyl-ether) mixed with quartz sand DW = dry weight Conc. = concentration REFERENCE ITEM No reference item is recommended for this test according to the test guideline. CONTROL 8 replicates applied with quartz sand. SOLVENT CONTROL 8 replicates applied with quartz sand and ferf.-methyl-buthyl- ether. Amount solvent added per treatment: oats: 63.1 mL soybean: 30 mL rape: 10 mL TEST CONDITIONS Duration A: oats Incorporation of the quartz sand into the soil: 2002- 11-2 1 Sowing of the seeds: 2002- 11-22 Completion date: 2002-12-13 B:soybean Incorporation of the quartz sand into the soil: 2002-11-28 Sowing of the seeds: 2002-11-29 Completion date: 2002- 12-20 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 15 of 51 Project-No. 020719CK Study-No. TNC84113 Number of replicates Number of seeds Test medium Test container Sowing Application Application technique C: rape Incorporation of the quartz sand into the soil: 2003-01-16 Sowing of the seeds: 2003-01-17 Completion date: 2003-02-07 8 replicates per dose level, solvent control and control were tested in a randomized block design. 5 seeds were sown per replicate. 2/3 certified LUFA soil No. 2.2 and V3 quartz sand (Non-GLP data) Overall carbon content: 1.31 % Certified LUFA soil No. 2.2 (batch-no. Sp 223302 / 224602) (Non-GLP data) Soil type: loamy sand (DIN) Grain size: < 2 mm Carbon content: 2.3 0.2 % pH value 5.6 0.4 The soil has been stored at the test facility at green house conditions until use. Origin: La n d w ir t s c h a f t l ic h e U n t e r s u c h u n g s - u n d Fo r s c h u n g s a n s t a l t S p e y e r (LUFA), Obere Langgasse 40, D-67346 Speyer. Plastic containers (standard flower pots) with a diameter of 12 cm were used. Test containers were filled with 700 g treated soil each (see below) and seeding holes were made with a seeding pistil. One seed was placed in every hole and covered with soil. Dur ing cultivation the test containers were bottom watered and fertilized as necessary. The control, solvent control and test item were applied once at the beginning of the study. To reach a sorption equilibrium the test item was incorporated into the soil 24 h prior to test start. The concentrations of the test item are expressed in terms of the dry weight of the soil. 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 16 of 51 Project-No. 020719CK Study-No. TNC84113 The test item was weighed out and dissolved in ferf.-buthylmethyl-ether to produce a number of stock solutions. A speci fied amount of the appropriate test item stock solution was mixed with quartz sand. After the solvent had volatilised out of the quartz sand, the treated quartz sand was mixed with the soil. The soil was mixed carefully with a hand-mixer (Kr u p s ) to insure a homogeneous distribution of the test item in the soil. The amount of quartz sand added was the same for each test concentration (1 %). A fter 24 h the treated soil was placed in the test container and the seeds were sown (see above). ENVIRONMENTAL CONDITIONS Source of light Watering / Fertilisation Nutrient solution nominal Oats Soybean Rape Temperature [C] 20 t 10 16 - 24 16 - 24 17 - 26 Relative humidity [%] > 60 68 - 100 70 - 100 74 - 100 Photoperiod [h] 16 16 16 16 Illumination [lx] > 3500 3600t 1100 5300t 1400 3400t 1100 Special-fluorescent tube Flu o r a (Art-No. 25414925) Special-fluorescent tube Da y l ig h t (Art.-No. 25414922) Origin: Ho k la t h e r m GMBH, Postfach 206, D-26687 Apen Bottom watering of the test containers with nutrient solution as necessary. Flory 9 hydro (0.5 %o) Origin: g r t n e r e ib e d a r f A s p e r g E.G., Niederlassung Han nover, Ringstrasse 7, D-30457 Hannover Nutrient Ammonium nitrogen Nitrate nitrogen P2 O5 Potassium oxide Magnesium oxide Boron Iron (Chelate) Copper Manganese Molybdenum Zinc Concentration [mg/L] 50 100 70 220 60 0.3 1.2 0.02 0.5 0.05 0.1 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 17 of 51 Project-No. 020719CK Study-No. TNC84113 TYPE AND FREQUENCY OF MEASUREMENTS AND OBSERVATIONS The room temperature and relative humidity were recorded continuously throughout the test with a thermohygrograph. During the observation period the replicates were observed every 7 days for number of emerged seedlings, number of dead plants and visual phytotoxic effects. Observations included all variations, either inhibitory or stim u latory, between the treated replicates and the pooled controls. Such variations were phytotoxic symptoms (e.g. chlorosis, ne crosis, wilting), formative effects and growth and development rates. At the end of the study the number of emerged seedlings, number of dead plants and phytotoxic effects were determined and shoot height and shoot fresh weight were measured. TEST DESCRIPTION Day -1: Start (day 0): Every working day: Day 7, 14, 21: Test end: A preliminary NON-GLP test was performed with 3 soil concentrations (10 - 100 - 1000 mg/kg DW), and additionally for rape a single concentration of 1 mg/kg DW was tested. Based on the results of the preliminary test, the main study was performed with five concentrations from 10 to 1000 mg/kg DW with a factor of V10 (oats and soybean) and from 6.25 to 100 mg/kg DW with a factor of 2 (rape). The test medium was heated to 50 C, weighed out, dissolved in fe/f.-buthyl-m ethyl-ether and mixed with quartz sand. After the solvent had volatilised out of the quartz sand (ca. 1 h), the treated quartz sand was mixed with the soil. The treated soil was placed in the test containers and the seeds were sown. Test containers were bottom watered and fertilised as needed. The plants were observed for number of emerged seedlings, visual phytotoxic effects and number of dead plants. Final shoot heights and fresh weight measurements were done. 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 18 of 51 Project-No. 020719CK Study-No. TNC84113 STATISTICS AND CALCULATIONS NOEC of biomass/ be emerged seedlings One Way Analysis of Variance (ANOVA) and Dunnett's Test were carried out for the determination of statistically signifi cant differences compared to pooled control replicates. When running a One Way Analysis of Variance a Normality Test and an Equal Variance Test were done first. P-values for both Normality and Equal Variance Test are 0.05. The a-value for ANOVA and Dunnett's Test (acceptable probability of incorrectly concluding that there is a difference) is a=0.05. No statistically significant differences were found between the control and the solvent control. Therefore for calculations the replicates of the control and solvent control were summarized in a pooled control group. All dead plants were measured and weighed and included in calculations if possible according to the guideline. EC-values and confidence Intervaly of growth Calculations were carried out via probit analysis. Probit values were taken from W eber (1986). Confidence intervals were calculated according to a standard procedure (Breitig & T mpling 1982). Software SigmaPlot (windows) rel. 6.0 (2000), SPSS Incorporation SigmaStat (windows) rel. 2.03 (1997), SPSS Incorporation Excel rel. 9.0 (2000), Microsoft Corporation VALIDITY CRITERIA The following validity criteria must be found in the control rep licates: - a minimum of 65 % emergence - the mean seedling growth should not exhibit visible phytotoxic effects - the mean survival of the plants is at least 90 % at the end of the test DEVIATIONS FROM THE GUIDELINE None 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 19 of 51 Project-No. 020719CK Study-No. TNC84113* DEVIATIONS FROM THE STUDY PLAN REPORTING ARCHIVING 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 regula tions: 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 item Microfilms will be retained in a safe-deposit by Volksbank Sarstedt, 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 20 of 51 Project-No. 020719CK Study-No. TNC84113 4 Results No statistically significant differences were found between the control and the solvent con trol. Therefore for calculations the replicates of the control and solvent control were summarized in a pooled control group. 4.1 Visual Phytotoxicity The plants were observed on day 7, 14 and 21 for visual phytotoxic effects, number of dead plants and emerged seedlings. Observations are given in Table 19 - 21 (day 7), Table 22 24 (day 14) and Table 25 - 27 (day 21). Number of emerged seedlings is given in Table 10 12 (day 7), 13 - 15 (day 14) and 16 - 18 (day 21). Phytotoxic effects and rate of emergence after 21 days are given in Table 4 - 6. Table 4: Oats: Phytotoxic Effects and Rate of Emergence after 21 Days Phytotoxic effects Conc. Phytotoxic effects Replicate [mg/kg DW] 12345678 Pooled control 10 32 100 320 1000 - - - ------- ------- ------- ------- ------- ------- ------- Conc. = Concentration - = not occurred Rate of Emergence [% ] 88 88 90 85 68 78 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 21 of 51 Project-No. 020719CK Study-No. TNC84113 Table 5: Rape: Phytotoxic Effects and Rate of Emergence after 21 Days Phytotoxic effects Conc. Phytotoxic effects* Replicate [mg/kg DW] 123456 Pooled control 6.25 12.5 25 50 100 - (E) (2.1) - ----- ----- ----- ----- ----- ----++ ++ + ++ ++ + Rate of Emergence 78 [%] ----1 --+ ++ 99 95 98 95 95 98 Table 6: Soybean: Phytotoxic Effects and Rate of Emergence after 21 Days Phytotoxic effects Conc. Phytotoxic effects* Replicate Rate of Emergence [mg/kg DW] 1 2 34 56 7 8 [% ] Pooled control 10 32 100 320 1000 (E) (E) (E) - - - - - 11- - ------- - - 1- - - - - - - 1- - - ------- ------- ------- 93 93 85 90 95 80 Conc. = Concentration * The numbers in brackets correspond to the observations as follows: (2.1) : Decrease in length (E) : Dead plant - = not occurred +/- = slight effect + = medium effect ++ = strong effect 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 22 of 51 Project-No. 020719CK Study-No. TNC84113 4.2 Shoot Height The shoot height was determined on day 21. Test item related statistically significant inhibition (marked with *) of the shoot heights was found at the concentration 100 mg/kg DW for rape (Figure 2). No test item related inhibition was determined in the tested concentration range of 10 - 1000 mg/kg DW for the plant species oats and soybean (Figure 1, 3). concentration [mg/kg DW] Figure 1: Oats: Shoot Height after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil Figure 2: Rape: Shoot Height after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 23 of 51 Project-No. 020719CK Study-No. TNC84113 Figure 3: Soybean: Shoot Height after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil 4.3 Fresh Weight (Biomass) The biomass was determined as fresh weight on day 21. Test item related statistically significant inhibition (marked with *) of the fresh weight was found at concentrations > 50 mg/kg DW for rape and at the concentration 1000 mg/kg DW for soybean (Figure 5, 6). No test item related statistically significant inhibition of the fresh weight was determined in the tested concentration range 10 - 1000 mg/kg DW for the plant species oats (Figure 4). Figure 4: Oats: Fresh Weight after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 24 of 51 Project-No. 020719CK Study-No. TNC84113 Figure 5: Rape: Fresh Weight after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil Figure 6: Soybean: Fresh Weight after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 25 of 51 Project-No. 020719CK Study-No. TNC84113 4.4 Emerged Seedlings No statistically significant decrease was found for the plant species oats and soybean at the concentration range 10 - 1000 mg/kg DW and for the plant species rape at the concentration range 6.25 - 100 mg/kg DW (Figure 7 - 9). Figure 7: Oats: Emerged Seedlings after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil Figure 8: Rape: Emerged Seedlings after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 26 of 51 Project-No. 020719CK Study-No. TNC84113 Figure 9: Soybean: Emerged Seedlings after 21 Days (Mean SD) in Relation to the Test Item Concentration in Soil 4.5 Effect Levels 4.5.1 Inhibition of Shoot Height, Fresh W eight and Number of Emerged Seedlings The tabulated results represent rounded results calculated on the exact raw data. Table 7: Inhibition of Shoot Height C oncentration [m g/kg DW] Oats Pooled control 34.4 Shoot height [cmn] Inhib. Soy [%] bean - 20.2 C oncentration Inhib. [m g/kg DW ] Shoot height [cmn] Rape Inhib. [%] [%] - pooled co n tro l 10.2 - 10 37.0 -8 21.8 -8 6.25 10.8 -6 32 35.4 -3 20.6 -2 12.5 9.8 4 100 34.7 -1 21.2 -5 25 10.7 -5 320 34.2 0 20.6 -2 50 9.4 8 1000 32.4 6 19.1 6 100 6.4* 37 Inhib. = Inhibition negative values = enhanced growth * Significantly different, P < 0.05, ANOVA a = 0.05 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 27 of 51 Project-No. 020719CK Study-No. TNC84113 Table 8: Inhibition of Fresh Weight C oncentration [m g/kg DW] Oats Pooled control 10 32 100 320 1000 816 979 917 795 759 718 Fresh weight [m g] Inhib. Soy [%] bean - 1792 -20 1737 -12 1614 3 1750 7 1553 12 1452 C oncentration Inhib. [m g/kg DW ] Fresh weight [m g] Rape Inhib. [%] [%] - pooled control 1035 - 3 6.25 1081 -4 10 12.5 975 6 2 25 1001 3 13 50 798* 23 19* 100 287* 72 Table 9: Inhibition of Emerged Seedlings C oncentration [m g/kg DW] Erm erged seedling s Oats [% ] Inhib. Soy [%] bean Inhib. [% ] C oncentration [m g/kg DW] Eme 'ged seedlin gs [%] Rape Inhib. [% ] P ooled co n tro l 88 - 93 - pooled control 99 - 10 88 0 93 0 6.25 95 4 32 90 -2 85 9 12.5 98 1 100 85 3 90 3 25 95 4 320 68 23 95 -2 50 95 4 1000 78 11 80 14 100 98 1 Inhib. = Inhibition negative values = enhanced growth * Significantly different, P < 0.05, ANOVA a = 0.05 4.5.2 EC2 5 , EC50 and NOEC Values EC25 and EC50 values with confidence range of shoot height, fresh weight and emerged seedlings were calculated out of percentage inhibition. Results were presented in Table 1-3. NOEC values were calculated using One W ay Analysis of Variance (An o v a ) and Du n n e t t 's test. Results were also presented in Table 1-3. 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 28 of 51 Project-No. 020719CK Study-No. TNC84113 5 Graphs Figure 10: Rape: Shoot Height after 21 d F igure 11: Rape: Fresh W eight after 21 d 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 29 of 51 Project-No. 020719CK Study-No. TNC84113 6 Conclusions A fter 21 days no test item related phytotoxic effects were observed for the plant species oats and soybean at the tested concentration range 10 - 1000 mg/kg DW. For rape decrease in length was observed at the concentration 100 mg/kg DW as medium to strong effect. For details see Table 25 - 27. The EC50-value for shoot height was not calculable in the tested concentration range for the tested plant species because no test item related inhibition occurred. The EC50- values for shoot height were > 100 mg/kg DW for rape and > 1000 mg/kg DW for oats and soybean, respectively (Table 1). Significant differences in comparison to the control were determined at the concentration 100 mg/kg for rape. The EC50- values for fresh weight was determined for the plant species rape at 73.7 mg/kg DW (Table 2). The EC50- values for fresh weight were > 1000 mg/kg DW for oats and soybean (Table 2). Significant differences in comparison to the control were determined at the concentration 50 and 100 mg/kg for rape and at the concentration 1000 mg/kg for soybean. The EC50- value for seedling emergence was not calculable in the tested concentration range for the tested plant species. The EC50- values for seedling emergence were > 100 mg/kg DW for rape and > 1000 mg/kg DW for oats and soybean, respectively (Table 3). 7 Validity Criteria The control treatm ent validity criteria according to the test guideline were fullfilled: - a minimum of 65 % emergence - the mean seedling growth exhibited no visible phytotoxic effects - the mean survival of the plants was at least 90 % at the end of the test 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 30 of 51 Project-No. 020719CK Study-No. TNC84113 8 Literature (1) Br e it ig & T m p l in g (1982): Methoden der W asseruntersuchungen Bd.II, VEB Gu s t a v Fis c h e r V e r l a g , Jena. (2) DuPont, Determination of physio-chemical parameters of Perfluorooctylethanol, CAS No. 678-39-7, Unpublished results, Update Feb. 15, 2002 (3) F l l C., J u n g S. u n d C. Sc h u l t e (1999): Prfanforderungen des Umweltbundesamtes zur Bewertung der Auswirkungen von Pflanzenschutzmitteln auf terrestrische Pflan zen. UWSF - Z. Umweltchem. kotox. 11 (3) 145-149 (4) OECD 208 (draft document, July, 2000), Terrestrial (Non-Target) Plant Testing, Part A Seedling Emergence Test (5) W e b e r . E.: Grundri der biologischen Statistik, 9. Auflage (1986). 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 31 of 51 Project-No. 020719CK Study-No. TNC84113 9 Biological Data 9.1 Observation Parameter (2) Disturbance of growth (2.1) Decrease in length (2.2) Promoted growth (3) (3.1) (3.2) (3.3) (3.4) Alteration of form and appearance (deformation) Curling / twisting / sticking or other deformation of the leafs Change of appearance Shoot deformations W ilting (4) (4.1) (4.3) (4.4) Discolouration of the leafs Lighter green (4.2) Chlorosis (4.3.1) (4.3.2) (4.3.3) (4.3.4) Loss of colour or discolouration darker green spots marginal of the veins between the veins (5) (5.1) Alteration of the leaf cells Necrosis (5.1.1) (5.1.2) (5.1.3) (5.1.4) (5.1.5) spots marginal apical between the veins of the whole leaf (E) Dead plant Assessment of the effects: - = no effects observed +/- = slight effects + = medium effects ++ = strong effects 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 32 of 51 Project-No. 020719CK Study-No. TNC84113 9.2 Emerged Seedlings on Day 7 Table 10: Oats: Emerged Seedlings on Day 7 Emerged seedlings Concentration Emergence Replicate [mg/kg DW] [% ] 12 34 56 7 8 Pooled control 10 32 100 320 1000 85 83 90 85 65 70 54543434 5 54 53 54 5 5 54 4 5 34 3 5 54 534 5 5 53533555 4 3442 153 34333255 Table 11: Rape: Emerged Seedlings on Day 7 Emerged seedlings Concentration Emergence Replicate [mg/kg DW] [% ] 12 34 56 7 8 Pooled control 96 54555555 55555535 6.25 93 5 3 4 5 5 5 5 5 12.5 95 5 5 5 5 5 4 5 4 25 95 5 5 5 4 5 4 5 5 50 95 4 5 5 5 5 4 5 5 100 95 5 5 5 5 4 5 5 4 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 33 of 51 Project-No. 020719CK Study-No. TNC84113 Table 12: Soybean: Emerged Seedlings on Day 7 Emerged seedlings Concentration Emergence Replicate [mg/kg DW] [% ] 12 34 56 7 8 Pooled control 10 32 100 320 1000 83 83 80 83 90 78 4 35 54 554 34335544 54544 155 2 54 35 54 4 4 54 54 34 4 5 34 55 554 4 32 54 55 3 9.3 Emerged Seedlings on Day 14 Table 13: Oats: Emerged Seedlings on Day 14 Emerged seedlings Concentration Emergence Replicate [mg/kg DW] [% ] 12 34 56 7 8 Pooled control 85 54543434 5 54 53 54 5 10 88 5 5 4 4 5 4 4 4 32 90 5 5 4 5 3 4 5 5 100 85 5 3 5 3 3 5 5 5 320 68 4 3 4 4 2 2 5 3 1000 78 4 4 3 4 3 3 5 5 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 34 of 51 Project-No. 020719CK Study-No. TNC84113 Table 14: Rape: Emerged Seedlings on Day 14 Emerged seedlings Concentration Emergence Replicate [mg/kg DW] [% ] 12 34 56 78 Pooled control 6.25 12.5 25 50 100 98 95 98 95 95 98 55555555 55555535 53555555 55 55 54 55 55545455 45555455 55555554 Table 15: Soybean: Emerged Seedlings on Day 14 Emerged seedlings Concentration Emergence Replicate [mg/kg DW] [% ] 12 34 56 7 8 Pooled control 91 44555554 4 54 55 54 4 10 93 5 4 5 5 5 3 5 5 32 85 2 5 4 3 5 5 5 5 100 90 5 5 4 5 5 4 4 4 320 95 5 4 4 5 5 5 5 5 1000 80 4 4 2 5 4 5 5 3 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 35 of 51 Project-No. 020719CK Study-No. TNC84113 9.4 Emerged Seedlings on Day 21 Table 16: Oats: Emerged Seedlings on Day 21 Emerged seedlings Concentration Emergence Sig. Replicate [mg/kg DW] [% ] Dif. 1 2 3 4 5 6 7 8 Pooled control 10 32 100 320 1000 88 88 90 85 68 78 54 54 34 3 4 55553555 No 5 5 4 4 5 4 4 4 No 5 5 4 5 3 4 5 5 No 5 3 5 3 3 5 5 5 No 4 3 4 4 2 2 5 3 No 4 4 3 4 3 3 5 5 Table 17: Rape: Emerged Seedlings on Day 21 Emerged seedlings Concentration Emergence Sig. Replicate [mg/kg DW] [% ] Dif. 1 2 3 4 5 6 7 8 Pooled control 99 55555555 55555545 6.25 95 No 5 3 5 5 5 5 5 5 12.5 98 No 5 5 5 5 5 4 5 5 25 95 No 5 5 5 4 5 4 5 5 50 95 No 4 5 5 5 5 4 5 5 100 98 No 5 5 5 5 5 5 5 4 Significantly different, ANOVA, a = 0.05 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 36 of 51 Project-No. 020719CK Study-No. TNC84113 Table 18: Soybean: Emerged Seedlings on Day 21 Emerged seedlings Concentration Emergence Sig. Replicate [mg/kg DW] [% ] Dif. 1 2 3 4 5 6 7 8 Pooled control 10 32 100 320 1000 93 93 85 90 95 80 44555554 45455554 No 5 4 5 5 5 3 5 5 No 2 5 4 3 5 5 5 5 No 5 5 4 5 5 4 4 4 No 5 4 4 5 5 5 5 5 No 4 4 2 5 4 5 5 3 Significantly different, ANOVA, a = 0.05 9.5 Visual Phytotoxicity on Day 7 Table 19: Oats: Visual Observations and Number of Dead Plants on Day 7 Phytotox ic effec ts Concentration Phytotoxic Rep icate [mg/kg DW] effects 1 2 3 4 5 6 7 8 Pooled control 10 32 100 320 1000 - - -------------------------------------------------- - = not occurred 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 37 of 51 Project-No. 020719CK Study-No. TNC84113 Table 20: Rape: Visual Observations and Number of Dead Plants on Day 7 Phytotoxic effects Concentration Phytotoxic Replicate [mg/kg DW] effects 1 2 3 4 5 6 Pooled control 6.25 12.5 25 50 100 - - ------------------------------------ 7 - 8 - Table 21: Soybean: Visual Observations and Number of Dead Plants on Day 7 Phytotox ic effec ts Concentration Phytotoxic Rep icate [mg/kg DW] effects 1 2 3 4 5 6 7 Pooled control 10 32 100 320 1000 - - ------------------------------------------- - = not occurred 8 - 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 38 of 51 Project-No. 020719CK Study-No. TNC84113 9.6 Visual Phytotoxicity on Day 14 Table 22: Oats: Visual Observations and Number of Dead Plants on Day 14 Phytotoxic effects Concentration Phytotoxic Replicate [mg/kg DW] effects 1 2 3 4 5 6 Pooled control 10 32 100 320 1000 - - ------------------------------------ 7 - 8 - Table 23: Rape: Visual Observations and Number of Dead Plants on Day day 14 Phytotox ic effec ts Concentration Phytotoxic Rep icate [mg/kg DW] effects 1 2 3 4 5 6 7 Pooled control 6.25 12.5 25 50 100 - - ------------------------------------------- - = not occurred 8 - 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 39 of 51 Project-No. 020719CK Study-No. TNC84113 Table 24: Soybean: Visual Observations and Number of Dead Plants on Day 14 Phytotoxic effects Concentration Phytotoxic Replicate [mg/kg DW] effects* 1 2 3 4 5 6 7 Pooled control (E) 10 32 100 320 1000 (E) - - - - - 11------------- - - - 1- ------------------- 8 - 9.7 Visual Phytotoxicity on Day 21 Table 25: Oats: Visual Observations and Number of Dead Plants on Day 21 Phytotoxic effects Concentration Phytotoxic Replicate [mg/kg DW] effects 1 2 3 4 5 6 Pooled control 10 32 100 320 1000 - - ------------------------------------ * The numbers in brackets correspond to the observations as follows: (E) : Dead plants - = not occurred 7 - 8 - 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 40 of 51 Project-No. 020719CK Study-No. TNC84113 Table 26: Rape: Visual Observations and Number of Dead Plants on Day 21 Concentration Phytotoxic [mg/kg DW] effects* Pooled control - 6.25 12.5 25 50 100 (E) (2.1) 1 ++ 2 ++ Phytotox ic effec ts Rep icate 3456 ------------------+ ++ ++ + 7 + 8 1 ++ Table 27: Soybean: Visual Observations and Number of Dead Plants on Day 21 Phytotoxic effects Concentration Phytotoxic Replicate [mg/kg DW] effects* 1 2 3 4 5 6 Pooled control (E) ----1 ----- 10 (E) - - - 1 - 32 (E) - - - - 1 100 - - - - - - 320 - - - - - - 1000 - ----- * The numbers in brackets correspond to the observations as follows: (2.1) : Decrease in length (E) : Dead plant - = not occurred +/- = slight effect + = medium effect ++ = strong effect 1 - 7 - 8 - 9.8 Shoot Height Statistical analysis was carried out using ANOVA and DUNNETT's Method if applicable (ANOVA a = 0.05). Multiple Comparisons vs. Control Group were done. Results see table 28 - 33. 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 41 of 51 Project-No. 020719CK Study-No. TNC84113 Table 28 : Oats: Shoot Height [cm] Pooled Control, 10 and 32 mg/kg DW Repl. Pooled control Control Height [cm] Mean SD Solvent control Height [cm] Mean SD Height [cm] Concentration [mg/kg DW] 10 32 Height Mean SD [cm] Mean SD 1 2 3 4 5 6 7 8 Mean SD Sig. Dif. P< 0.05 4.9 40.6 37.5 38.4 34.8 38.7 36.2 32.9 39.7 37.0 39.8 33.6 43.6 44.2 42.2 35.6 34.8 38.4 37.7 31.5 35.0 32.5 35.3 36.0 36.4 36.1 35.4 33.9 29.9 37.9 17.4 24.7 - 31.2 14.9 36.9 3.0 39.6 4.5 37.8 3.3 34.7 3.1 35.1 1.8 35.1 1.1 27.5 8.6 37.4 37.0 38.4 37.7 30.2 38.5 41.3 36.0 38.7 33.9 37.5 39.8 39.0 32.6 3.4 38.4 33.6 35.4 33.4 33.3 37.8 40.6 34.1 36.1 37.1 39.4 38.6 33.9 3.6 29.8 32.0 37.9 36.3 32.0 32.6 27.0 33.5 34.6 34.5 3.6 36.1 3.4 37.7 2.8 30.5 15.4 34.8 2.2 37.5 3.3 37.0 2.2 27.9 14.0 31.9 2.9 42.9 38.6 36.7 39.1 36.3 37.8 43.0 40.4 36.8 36.8 38.3 37.4 39.6 38.4 48.7 46.7 39.5 42.6 33.8 32.6 39.0 41.9 36.0 9.3 35.8 41.3 43.5 16.7 38.1 41.6 36.2 7.3 38.4 43.3 42.4 - 38.7 2.6 39.0 2.7 38.4 0.9 44.4 4.1 36.7 3.8 32.5 15.8 33.2 11.2 32.9 17.2 37.0 4.1 No 3.4 35.6 38.8 38.0 33.4 35.1 37.7 36.6 34.5 34.5 35.5 33.4 32.7 37.4 41.8 43.6 41.5 34.1 44.4 33.7 36.7 41.0 34.3 36.1 37.1 43.6 36.3 34.1 29.6 35.9 35.0 39.7 34.9 32.9 38.1 19.3 29.8 14.9 35.7 1.4 34.8 2.1 41.1 4.1 37.1 3.7 37.8 4.1 34.2 2.7 33.0 8.1 35.4 3.4 No - not emerged Sig. Dif. = Significantly different 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 42 of 51 Project-No. 020719CK Study-No. TNC84113 Table 29 : Oats: Shoot Height [cm] 100, 320 and 1000 mg/kg DW Repl. 1 2 Concentration [mg/kg DW] Height [cm] 100 Mean SD Height [cm] 320 Mean SD Height [cm] 1000 Mean SD 34.0 36.3 32.5 30.0 34.4 42.8 29.3 38.2 - 33.4 2.4 36.8 6.9 31.4 32.0 28.8 37.9 - 35.1 32.2 33.2 - 32.5 3.8 33.5 1.5 34.9 25.9 35.3 32.2 - 38.0 36.7 35.9 30.6 32.1 4.3 35.3 3.3 35.8 36.0 39.2 35.2 37.0 32.7 3 36.9 35.7 3.3 39.2 36.6 2.0 33.7 35.2 3.5 30.9 34.3 - 39.9 - - 35.1 31.4 27.4 33.1 33.1 38.1 4 35.6 34.6 1.3 33.3 32.8 0.9 26.9 31.8 5.5 - 33.3 34.8 32.7 37.1 35.9 34.8 36.7 31.8 5 38.2 35.2 2.8 36.9 0.3 17.9 28.5 9.4 - 6 7 8 Mean SD Sig. Dif. P< 0.05 34.3 33.0 36.0 32.0 35.0 36.6 36.1 25.6 37.3 29.1 38.6 34.1 39.6 33.9 26.7 34.1 1.6 32.9 5.3 34.6 5.1 34.7 1.3 No - not emerged Sig. Dif. = Significantly different 26.4 35.7 37.9 30.0 35.1 36.3 37.4 35.4 34.0 36.0 - 31.1 6.6 35.3 3.2 35.1 1.0 34.2 2.1 No 33.6 27.6 29.3 35.7 39.8 29.4 35.7 32.9 32.8 19.2 35.2 37.0 33.6 30.2 3.1 34.7 3.9 31.6 7.1 32.4 2.5 No 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 43 of 51 Project-No. 020719CK Study-No. TNC84113 Table 30 : Rape: Shoot Height [cm] Pooled Control, 6.25 and 12.5 mg/kg DW Pooled control Repl. Control Height [cm] Mean SD Solvent control Height [cm] Mean SD 1 2 3 4 5 6 7 8 Mean SD Sig. Dif. P< 0.05 11.5 8.7 9.9 5.7 9.4 9.4 9.3 11.2 9.5 2.5 13.5 10.8 10.2 10.9 9.1 11.3 9.2 9.1 10.0 10.8 10.3 10.9 8.1 10.9 4.7 8.6 7.6 8.5 9.5 8.6 10.5 9.4 9.2 10.1 11.1 8.3 9.3 9.9 7.8 10.5 9.0 2.1 8.4 3.4 10.9 1.6 10.1 1.0 9.0 2.7 8.6 0.7 10.1 0.8 9.2 1.1 10.8 8.9 10.4 10.7 11.3 12.0 9.4 12.2 12.1 10.7 10.3 13.3 11.6 9.7 10.0 13.2 12.9 12.6 10.4 11.8 11.2 11.5 11.1 13.1 11.1 10.8 10.8 11.1 10.9 8.4 11.9 13.0 10.5 3.4 - 11.2 11.5 11.6 10.5 11.2 10.2 1.1 10.4 0.9 11.3 1.2 11.0 1.5 12.2 1.1 11.6 0.9 10.4 1.1 9.7 4.3 11.2 0.4 - not emerged Sig. Dif. = Significantly different Concentration [mg/kg DW] 6.25 Height [cm] Mean SD Height [cm] 12.5 Mean SD 11.6 12.0 11.3 11.4 10.7 11.6 11.9 9.2 - - 9.7 11.9 12.4 3.9 11.2 11.3 11.0 11.1 11.6 10.8 14.1 10.6 11.0 11.6 9.3 11.7 11.4 9.2 10.5 10.5 11.7 10.2 10.8 11.2 11.2 11.3 7.8 10.2 10.6 10.1 11.4 0.5 10.9 1.5 9.8 3.5 11.2 0.3 11.3 1.8 10.7 1.0 11.0 0.6 10.0 1.3 10.7 10.8 9.6 10.2 10.3 11.6 13.4 10.9 11.1 12.3 9.7 11.8 9.8 8.9 10.5 7.4 11.1 9.8 9.7 9.8 8.9 9.0 10.1 8.9 10.8 8.4 11.2 8.7 9.4 - 10.9 10.6 10.8 11.1 10.2 2.5 9.1 5.4 8.2 7.4 10.3 0.5 11.9 1.0 10.1 1.1 9.6 1.3 9.5 0.9 9.4 1.3 10.7 0.3 6.5 2.6 10.8 0.6 9.8 1.5 No No 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 44 of 51 Project-No. 020719CK Study-No. TNC84113 Table 31 : Rape: Shoot Height [cm] 25, 50 and 100 mg/kg DW Concentration [mg/kg DW] Repl. Height [cm] 25 Mean SD Height [cm] 50 Mean SD Height [cm] 100 Mean SD 1 2 3 4 5 6 7 8 Mean SD Sig. Dif. P< 0.05 10.3 5.2 11.0 12.4 7.3 11.4 12.6 10.1 12.8 12.9 11.0 10.7 9.8 9.5 10.3 11.3 11.3 11.0 12.2 - 11.5 10.2 10.6 10.8 10.6 12.0 11.4 10.9 10.0 - 9.3 10.4 9.8 10.8 11.2 11.5 12.4 10.3 9.5 9.9 9.2 2.9 12.0 1.2 10.3 0.6 11.5 0.5 10.7 0.5 11.1 0.8 10.3 0.8 10.7 1.2 10.7 0.8 No 10.8 7.6 9.6 9.3 - 10.8 11.0 7.0 11.5 8.0 8.7 10.9 11.1 10.5 10.3 8.3 7.4 8.8 9.3 6.5 11.7 9.2 10.3 9.0 10.0 8.5 11.0 11.1 9.3 - 10.0 9.4 9.2 5.5 9.5 10.2 8.7 10.3 10.8 6.1 9.3 1.3 9.7 2.0 10.3 0.9 8.1 1.1 10.0 1.1 10.0 1.3 8.7 1.8 9.2 1.9 9.4 0.7 No 5.1 6.3 7.5 5.8 1.2 4.5 5.5 8.1 3.2 5.5 5.5 2.1 7.1 3.7 7.4 4.7 9.4 7.1 1.8 8.1 6.1 3.8 4.9 5.5 5.0 0.9 6.1 4.9 8.7 6.8 7.2 7.0 1.8 8.3 4.1 7.9 5.0 5.7 6.1 1.5 7.5 4.4 8.7 6.3 7.2 7.8 1.1 8.9 7.7 6.6 6.6 7.7 6.6 1.0 5.3 - 6.4 0.9 Yes - not emerged Sig. Dif. = Significantly different 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 45 of 51 Project-No. 020719CK Study-No. TNC84113* Table 32 : Soybean: Shoot Height [cm] Pooled Control, 10 and 32 mg/kg DW Repl. Pooled control Control Height [cm] Mean SD Solvent control Height [cm] Mean SD Height [cm] Concentration [mg/kg DW] 10 32 Height Mean SD [cm] Mean SD 1 2 3 4 5 6 7 8 Mean SD Sig. Dif. P< 0.05 20.6 19.2 22.1 23.2 - 18.7 21.6 22.5 12.7 19.5 21.2 24.1 25.5 16.7 4.0 4.5 22.8 23.8 21.7 12.0 24.0 22.7 24.5 * 25.8 24.3 23.1 23.6 * 24.4 23.6 23.9 23.8 22.3 6.2 23.6 25.3 25.1 - 21.3 1.7 18.9 4.4 21.4 3.5 15.4 10.2 20.8 5.9 24.2 1.2 23.6 0.8 20.1 9.3 22.3 25.6 18.8 18.0 23.5 23.1 24.7 23.7 21.1 22.7 8.7 9.8 22.7 - 4.5 11.1 24.4 25.2 21.9 8.5 21.7 23.1 19.7 23.8 22.7 3.2 22.1 23.9 21.1 24.0 23.0 22.3 22.2 12.8 23.7 23.9 24.2 15.9 - 20.3 2.5 21.2 3.5 23.2 1.3 16.0 7.8 17.4 9.2 19.4 6.3 18.6 8.7 20.9 4.6 21.9 4.0 12.0 23.8 20.9 18.5 23.1 21.5 18.1 21.4 21.6 25.2 15.5 27.0 23.4 19.2 23.0 23.5 25.6 27.5 * 19.7 4.8 20.7 1.7 22.1 4.7 24.9 2.1 25.9 24.4 18.8 23.9 23.4 16.1 16.7 23.3 17.9 22.6 24.3 23.4 25.3 21.4 24.7 21.6 24.2 20.9 23.3 2.7 18.7 4.0 22.7 2.9 22.6 1.8 21.8 2.0 No 16.8 21.3 16.9 15.1 22.0 23.8 22.6 22.2 20.3 22.0 22.9 - 24.5 22.2 23.5 - 3.2 2.9 25.5 24.2 22.4 20.6 22.3 23.3 20.1 24.0 24.4 24.6 23.5 23.3 13.7 21.8 11.9 20.1 23.6 25.2 19.1 3.2 20.1 3.8 21.9 1.1 23.4 1.2 15.6 11.5 22.1 1.7 21.9 4.6 20.5 5.2 20.6 2.4 No * not determinable, plant decomposed - not emerged Sig. Dif. = Significantly different 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 46 of 51 Project-No. 020719CK Study-No. TNC84113* Table 33 : Soybean: Shoot Height [cm] 100, 320 and 1000 mg/kg DW Repl. 1 2 3 Concentration [mg/kg DW] Height [cm] 100 Mean SD Height [cm] 320 Mean SD Height [cm] 1000 Mean SD 3.9 25.6 23.1 22.7 23.2 7.7 23.8 20.9 22.5 22.1 23.1 21.9 21.1 23.4 19.7 8.9 19.4 6.6 22.4 1.1 22.0 24.6 22.2 23.7 21.1 19.8 20.4 20.3 22.4 - 17.0 7.4 22.2 18.7 22.7 1.4 20.7 1.1 16.3 6.3 18.4 20.3 11.5 21.5 - 16.7 22.1 15.4 21.1 - 22.8 21.4 - - 17.9 4.5 18.8 3.3 22.1 1.0 4 5 6 7 8 Mean SD Sig. Dif. P< 0.05 25.5 23.4 24.4 24.7 25.5 24.4 24.0 16.2 20.8 22.8 10.0 15.1 13.1 25.0 - 20.4 22.8 22.6 23.0 - 23.2 22.6 24.5 23.6 - 24.7 0.9 21.6 3.3 15.8 6.5 22.2 1.2 23.5 0.8 21.2 2.8 No 21.3 20.1 23.9 15.0 23.1 21.9 20.7 23.9 26.2 23.7 4.1 24.0 22.0 20.3 23.7 21.5 23.3 17.8 22.7 22.9 18.9 22.8 23.3 18.4 19.5 20.7 3.5 23.3 2.1 18.8 8.4 21.6 2.2 20.6 2.3 20.6 2.2 No * not determinable, plant decomposed - not emerged Sig. Dif. = Significantly different 21.0 21.6 19.3 19.5 11.8 22.0 19.2 19.0 22.1 - 21.8 10.9 21.9 19.6 17.0 23.9 18.8 22.8 22.6 22.1 2.6 21.0 20.7 - - 18.6 3.9 20.6 1.7 18.2 4.6 22.0 1.9 14.8 10.5 19.1 2.4 No 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 47 of 51 Project-No. 020719CK Study-No. TNC84113 9.9 Fresh W eight (Biomass) Day 21* Table 34: Oats: Fresh Weight on Day 21 Conc. [m g /kg DW] Repl. No. Number of plants per replicate* Fresh w e ig h t mean value per replicate [m g ] Fresh w e ig h t mean value SD [m g ] Signif. Diff. Conc. Number of [m g /kg DW] plants per replicate* Fresh weight mean value per replicate [mg] o o o c o o ~o <D O O o CL = o o = > 10 32 1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8 5 4 5 4 3 4 3 4 5 5 5 5 3 5 5 5 5 5 4 4 5 4 4 4 5 5 4 5 3 4 5 5 765 1017 1092 1086 783 907 823 509 926 748 834 787 747 1002 572 465 1057 831 980 1318 1001 850 766 1030 795 858 879 1256 985 1054 757 749 100 320 817 189 1000 979 172 No 917 174 No 5 3 5 3 3 5 5 5 4 3 4 4 2 2 5 3 4 4 3 4 3 3 5 5 628 896 901 815 798 702 843 780 614 828 886 572 1134 601 630 804 576 790 1071 787 530 517 837 633 * number of plants per replicate included in calculations Conc. = Concentration SD = Standard deviation Signif. Diff. = Significantly different, ANOVA, a = 0.05. Fresh weight mean value SD [mg] 795 93 759 193 718 190 Signif. Diff. No No No 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 48 of 51 Project-No. 020719CK Study-No. TNC84113 Table 35: Rape: Fresh Weight on Day 21 Conc. [m g /kg DW] Repl. No. Number of plants per replicate* Fresh w e ig h t mean value per replicate [m g ] Fresh w e ig h t mean value SD [m g ] Signif. Diff. Conc. Number of [m g /kg DW] plants per replicate* Fresh weight mean value per replicate [mg] o o o c o o ~o <D O O o CL = o o = > 6.25 12.5 1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8 5 5 5 5 5 5 5 5 5 5 5 5 5 5 4 5 5 3 5 5 5 5 5 5 5 5 5 5 5 4 5 5 871 830 1160 1118 995 863 1339 848 918 1287 1103 1052 1202 817 1081 1071 1254 1259 1036 1232 1142 826 1022 878 965 1414 1031 765 1030 950 1232 416 25 50 1035 166 - 100 1081 169 No 975 298 No 5 5 5 4 5 4 5 5 4 5 5 5 5 4 5 5 5 5 5 5 5 5 5 4 885 1227 857 1184 907 1016 715 1213 768 950 834 459 829 1096 655 796 154 292 461 208 280 275 394 228 * number of plants per replicate included in calculations Conc. = Concentration SD = Standard deviation Signif. Diff. = Significantly different, ANOVA, a = 0.05. Fresh weight mean value SD [mg] 1001 191 798 189 287 100 Signif. Diff. No Yes Yes 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 49 of 51 Project-No. 020719CK Study-No. TNC84113 Table 36: Soybean: Fresh Weight on Day 21 Conc. [m g /kg DW] Repl. No. Number of plants per replicate* Fresh w e ig h t mean value per replicate [m g ] Fresh w e ig h t mean value SD [m g ] Signif. Diff. Conc. Number of [m g /kg DW] plants per replicate* Fresh weight mean value per replicate [mg] o o o c o o ~o <D O O o CL = o o = > 10 32 1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8 4 4 5 5 5 5 5 4 4 5 4 5 5 5 5 4 5 4 5 5 5 3 5 5 2 5 4 3 5 5 5 5 1755 1657 1701 1477 1393 1743 2003 1988 1856 2359 1441 1833 1747 1728 1952 2032 1792 1496 1838 1765 1703 1556 1745 2004 1414 1290 1332 2174 1234 1811 1908 1746 100 320 1792 246 1000 1737 159 No 1614 343 No 5 5 4 5 5 4 4 4 5 4 4 5 5 5 5 5 4 4 2 5 4 5 5 3 1616 1433 1921 1715 1720 1433 1910 2251 1656 1477 1252 1435 2075 1493 1671 1368 1146 1502 1755 1492 1624 1293 1579 1225 * number of plants per replicate included in calculations Conc. = Concentration SD = Standard deviation Signif. Diff. = Significantly different, ANOVA, a = 0.05. Fresh weight mean value SD [mg] 1750 274 1553 252 1452 211 Signif. Diff. No No Yes 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-heptadecafluoroTerrestrial Plants Toxicity, Seedling Emergence acc. to OECD Guideline 208 (Draft July 2000) Page 50 of 51 Project-No. 020719CK Study-No. TNC84113 10 Certificate of Test Item Analysis DR.U.NOACK-LABORATORIUM FR ANGEWANDTE BIOLOGIE Report Page 51 of 51 1-Decanol, 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-heptadecafluoro- Terrestrial Plants Toxicity, Seedling Emergence Project-No. 020719CK acc. to OECD Guideline 208 (Draft July 2000)_____________________Study-No. TNC84113 11 GLP Certificate of Dr .U.No a c k -Labo rato rium o Niederschsisches Umweltministerium Gute Laborpraxis/Good Laboratory Practice GLP-Bescheinigung/Statement of GLP Compliance (gem/according to 19 b Abs.1 Chemikaliengesetz) Eine GLP-Inspektion zur berwachung der Einhaltung der GLP-Grundstze gem Chemikaliengesetz bzw. Richtlinie 88/320/EG wurde durchgefhrt in: Assessment of conformity with GLP according to Chemikaliengesetz and Directive 88/320/E E C at: X Prfeinrichtung/Test facility Prfstandort / Test site Dr. U. NOACK LABORATORIUM fr angewandte Biologie Kthe-Paulus-Str. 1 D-31157 Sarstedt Dr. U. NOACK LABORATORY of applied biology Kthe-Paulus-Str. 1 D-31157 Sarstedt (Unverwechselbare Bezeichnung und Adresse/Unequivocal name and address) Prfungen nach Kategorien/Areas of Expertise (gem/according ChemVwV-GLP Nr. 5.3/O EC D guidance) 1 - Prfungen zur Bestimmung der physikalisch-chemischen Eigenschaften und Gehaltsbestimmungen 3 - Prfungen zur Bestimmung der erbgutverndernden Eigenschaften (in vitro und in vivo) 4 - Umwelttoxikologische Prfungen zu Auswirkungen auf aquatische und terrestrische Organismen 5 - Prfungen zum Verhalten im Boden, im Wasser und in der Luft, Prfungen zur Bioakkumulation und zur Metabolisierung 6 - Prfungen zur Bestimmung von Rckstnden 7 - Prfungen zur Bestimmung der Auswirkungen auf Mesokosmen und natrliche kosysteme Datum der Inspektion / Date of Inspection (Tag.Monat.Jahr / day.month.year) 25. - 27. Juni 2001 / June 25th- 27th, 2001 29. August und 21. September 2001 / August 29thand September 21*` , 2001 iDie genannte befindet sich im nationalen GLPberwachungsverfahren und wird regelmig auf Einhaltung der GLP-Grundstze berwacht. Auf der Grundlage des Inspektionsberichtes wird hiermit besttigt, dass in dieser Prfeinrichtung/diesem Prfstandort die oben genannten Prfungen unter Einhaltung der GLP-Grundstze durchgefhrt werden knnen. Niederschsisches Umweltministerium Referat 33 Archivstrae 2 30169 Hannover The above mentioned test facility / test site 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 / test- site is able to conduct the aforementioned studies in compliance with the Principles of GLP. 11. M rz 2002 Im Auftrge Dr. Braedt