Document ZBje3Zm2ZRG7z1ENyk7G5V6q8

3M Strategic Toxicology Laboratory Study Title: Acute Inhalation Toxicokinetic Study of Perfluoroctanesulfonyl Fluoride (POSF) T-7098.4 Test Number (sample): , T-7098.4 (Perfluoroctanesulfonyl Fluoride (POSF) Laboratory Project Identification: ST64 In-Life Initiation Date: 5/1/2001 In-Life Completion Date: 5/14/2001 Research Client: 3M Specialty Materials Division 3M Center Building 236 Saint Paul, MN 55133-3220 Testing Laboratory: 3M Strategic Toxicology Laboratory 3M Center Building 270-SB-181 Saint Paul, MN 55133-3220 Study Director: Andrew M. Seacat, Ph.D. Toxicologist Specialist 3M Medical Department/Toxicology Services 3M Center Building 220-2E-02 Saint Paul, MN 55133-3220 Study Toxicologist: Tommie Yvette Turner, Ph.D. Post Doctoral Research Fellow 3M Medical Department/Toxicology Services 3M Center Building 220-2E-02 Saint Paul, MN 55133-3220  T-7098.4 Acute Inhalation Toxicokinetic Study o f Perfluoroctanesulfonyl Fluoride (POSF) Final Report Summary: An acute inhalation toxicokinetic screen was conducted using 3 male rats for control (air-only) and 3 male rats exposed to perfluoroctanesulfonyl fluoride (POSF) at a nominal concentration of 1000 ppm for 4 hours. All animals gained weight during the recovery period and showed no clinical pathology. One animal (N=l) per time point from the POSF-treated animals was sacrificed on day 1, day 6 and day 14 post-exposure. All animals tolerated the 4-hour inhalation exposure and gained weight. Organ weights were normal and histological evaluation of the liver, lungs, bladder, kidney and brain revealed no significant findings. Perfluorooctanesulfonate (PFOS) was found in the serum from each animal (N=l) at 8.6,9.3 and 5.0 ppm of days 1, 6 and 14 post dose, respectively. The percentage of inhaled POSF represented by these serum PFOS concentrations were 0.18%, 0.23%, and 0. 10%, respectively. Conclusion: This study demonstrates that there was a small percentage (<0.5%) of total POSF inhaled found as PFOS in serum collected over 14 days. Results show that there was no significant change in PFOS in serum concentration over time. POSF can be absorbed from the lung, and metabolized to PFOS, which is present in serum for greater than 14 days. 1. Study Objective: This study was done to establish toxicokinetic parameters data on an acute inhalation of perfluoroctanesulfonyl fluoride (POSF). The goals of this study were to test the compound via inhalation in rats at 1000 ppm for 4 hours for distribution and metabolism, and to use the results as a guidance for a 13 week inhalation study. II. Test Articles: A. Identification: T# Name 7098'.3 POSF Structure MW (g/mole) Density (g/ml) C8F |7S 0 2F 502 1.69 B. Purity and Stability: Responsibility of research client. This POSF sample was lab fractionated and acid/water washed. Chemical characterization of C8F17SO2F by !HNMR and l9F-NMR Spectroscopy was conducted by 3M Specialty Adhesives & Chemicals Analytical Laboratory/ SMD to determine the purity o f the nominal product and to characterize as many impurity components as possible. The sample was found to be a high purity form of the nominal POSF product (1). C. Handling and Storage: Upon receipt, the test article was stored tightly sealed at room temperature. 2 T X-7098.4 Acule Inhalation Toxicokinetic Study o f Perfluoroctanesulfonyl Fluoride (POSF) Final Repon D. Disposition of Test Material: Any remaining unused portion of the test article was returned to the research client after completion of the study. H I. Test System: Male rats were exposed to test compound in air for 4 hours for one day in a 13 liter glass exposure chamber (3 rats/chamber/compound). The flow-through atmosphere was generated by mixing test compound from a syringe pump into a stream of fresh air flowing at 3.0 L/min into the chamber and exhausted through a vent. The syringe flow rate was calculated based on the Ideal Gas Law equation: ((target concentration)(MW)/(24.79L) x (lg/106pg) x (airflow rate) x (1/density) x (1000 pI/mL)) Given: Target concentration (ppm) MW = g/mole 24.79L: The volume of one mole of an ideal gas at 25 C and 1 atmosphere (760 mm Hg) conversion factor: (1 gram)/(l x 106|ig) airflow rate: (L/min) - density: (mL/g) conversion factor: 1000 pl/mL The target concentrations of each test compound, the syringe flow rate and the airflow rate were calculated using the Ideal Gas Law and are provided in the following table: Dose Group C Syringe Flow Rate Airflow Rate (ppm) (pl/min) (L/min) Treated: POSF 1000 30 2.0 - 3.0 Toxicokinetic group Control: Air only 2.0 - 3.0 Standard target concentrations for each compound were prepared and analyzed by Fourier Transform Infrared Spectrometry (FTIR). Chamber air was monitored by FTIR for concentrations of test compound and oxygen (2). Airflow was increased when necessary to maintain a level of at least 1000 ppm. Control animals were exposed to room air under equivalent chamber conditions as the POSF dose group animals. The control rats used in this study were also used for the control group in the limit test of POSF (T-7098.3) conducted at the same time. . Animals were handled in accordance with an animal usage protocol approved by LARC protocol # 2000-0068. The day of dosing was designated day zero of the study. Body weights were determined immediately prior to dosing and immediately prior to 3 -7098.4 Acute Inhalation Toxicokinetic Study o f Perfluoroctanesulfonyl Fluoride (POSF) Final Report euthanasia. At the termination of the study, all animals were humanely euthanized by CO2 asphyxiation and necropsies were performed. Blood serum was collected from one animal/time point of the toxicokinetic group sacrificed at day 1, day 6, and at day 14 post exposure for PFOS analysis. Analysis of PFOS levels by high-pressure liquid chromatography/tandem electrospray mass spectroscopy (HPLC-MS/MS) was performed according to published methods (3). Serum from the rat sacrificed day one post dose (Animal #R01468) was initially extracted and quantified by LC/MS/MS based on a PFOS standard curve extracted from urine collected from the limit toxicity study in rats (T-7098.3). This was done to get an estimate of serum PFOS levels on day one post-dose assuming an equal extraction of urine PFOS std curve and serum PFOS. Serum from this animal was later extracted and quantified against a standard curve extracted from serum, and therefore this data was not used in any calculations, but it is included here for completeness (Table 4). Lungs, liver, and bladder were collected at sacrifice. A portion of each tissue was immediately fixed in a 10% formalin solution for subsequent histological evaluation. The remaining sections were placed directly into propylene tubes filled with liquid nitrogen for possible future evaluation of POSF metabolites levels. IV. Results and Discussion: Test Atmosphere The POSF atmosphere generated in the exposure chamber, measured continuously for four hours by FTIR, averaged 932 93 ppm for the toxicokinetic group (4). The average measured concentration in the chamber was 93% of the target concentration for the toxicokinetic group. The airflow was increased from 2 to 3 liters/ minutes to adjust chamber concentration. Control Group: The three control group male rats had no adverse clinical observations noted during the 4 hr air-only exposure. The average body weight was 378 12 g at the initiation of the study. The control group gained an average o f 22 g (5.5%) of body weight during the 14 day study period. Liver weights averaged 14.1 1.0 g and the average liver to body weight ratio was 0.035 (Tablel). Necropsy findings and the histopathology report by Dr. Elden Lamprecht, 3M Surgical and Histopathology Services, is found in Appendix 1. All rat lungs (both control and treated groups) had mild to moderate congestion of alveoli with occasional extravasation of blood around arterioles and bronchioles. This is possibly an agonal change associated with CO2 euthanasia. It was noted that some of the lungs were not 4 T-7098.4 Acute Inhalation Toxicoidneuc Study o f Peril uoroctanesulfonyl Fluoride (POSF) Final Report well insufflated. Other than these artifacts, histological analysis revealed no significant changes in lungs and urinary bladder in all three of the controls. POSF Toxicokinetic Group: The three POSF group male rats showed no adverse clinical observations during the in-life phase of the rats. All animals gained weight during the study. At necropsy, no abnormal gross observations were made. Liver weights were equivalent to the control liver weights. Histological analysis in one of the animals revealed that one macroscopic focus of atelectasis (collapsed alveoli) was present in the lung lobe. The change appeared chronic in duration and was not considered to be compound-related. There were no significant changes in the bladder. At necropsy for each animal, there was a small percentage of POSF measured in the serum for each individual animal (Table 3). Perfluorooctanesulfonate (PFOS) was found in the serum from each animal (N =l) at 8.6, 9.3 and 5.0 ppm of days 1, 6 and 14 post dose, respectively. The percentage of inhaled POSF represented by these serum PFOS concentrations were 0.18%, 0.23%, and 0.10%, respectively. These data suggest that the serum PFOS concentrations were at or near their apparent peak concentrations for the first week following exposure and then PFOS began to clear from the serum over the following week. Although there is no statistical power in the present study, the data are consistent with a plasma elimination half-life of l4C following single oral administration of [14C]FC-95 (mean dose 4.2 mg/kg) to male rats of 7.5 days (5). Rat # R01468 was sacrificed on day 1 post dose and sera was collected and analyzed for PFOS. The calculation of serum PFOS was based on urine PFOS standard curve, assuming equal extraction (Figure 1). The amount of PFOS in serum averaged over time, collected on days 1, 6, and 14 post-exposure was 25.90 +_1.10 ppm (Table 4). Serum from this animal was later extracted and quantified against a standard curve extracted from serum. The data dervived from the PFOS standard curve (shown in table 4) extracted from urine was not used in any calculations, but it is included here for completeness. This toxicokinetic study confirmed early findings of a POSF 4-hour rat LCso greater than 1000 ppm in an inhalation limit toxicity study of POSF (6). In the limit study, rats were exposed to 1000 ppm POSF for four hours. All animals survived the duration of the study. The concentration of PFOS that was found in the serum on day 13 post -dose was 4.57 + 0.70 ppm (N=3) which represented approximately 0.10 0.01 % of the inhaled dose of POSF. The average amount of PFOS in the day-one overnight urine was 3.08 1.60 ppm (N=3) which represented approximately 0.05 0.03% on the inhaled dose of POSF (6). Thus, the data from the POSF limit test and the data from the toxicokinetic test presented herein are in agreement with each other and support the same conclusions. 5 tT-7098.4 Acute Inhalation Toxicokineuc Study o f Perfluoroctanesulfonyl Fluoride (POSF) Final'Report V. Conclusion: This study demonstrates that there was a small percentage (<0.5%) of total POSF inhaled found as PFOS in serum collected over 14 days. Results show that there was no significant change in POSF in serum concentration over time. 6 T-7098.4 Acute Inhalation Toxicokinetic Study o f Perfluoroctanesulfonyl Fluoride (POSF) Final' Report VI. References: 1. Kestner, T. Chemical Characterization of POSF (FM-3270) by 'H-NMR & I9F-NMR Spectroscopy. 3M Specialty Materials & Manufacturing Division Analytical Laboratory. Request No. 6370. May 15, 2001. 2. Modified EPA Method 320. Pace Analytical. 3. Hansen, K.J., Clemen, L.A., Ellefson, M.E. and Johnson, H.O. (2001) Compound Specific, Quantitative Characterization of Organic Fluorochemicals in Biological Matrices. Environ Sei and Tech., 35, 766-770. 4. Gutzkow, T. POSF Animal Exposure Test Report. Pace Analytical Services, Inc. 3M Laboratory Request No. E01-0648. 3M ET&SS Field Analytical Services and Technologies. 5. Johnson, J. D. and Ober, R.F . 1979. Absorption of FC-95-14C in rats after a single oral dose. Project No. 8900310200, Riker Laboratories, Inc., St. Paul, MN. 6. Sect, A. and Turner, T. (2001) Acute Inhalation Limit Toxicity Study of Perfluooctanesulfonyl Fluoride (POSF) T-7098.3. 7 T-7098.4 Acute Inhalation Toxicokinetic Study o f Perfluoroctanesulfonyl Fluoride (POSF) Final Report ,, Andrew Seacat, Ph.D. Toxicologist Specialist Study Director Reviewed by: John L. Butenhoffr Ph.D, C IH ,DABT Toxicologist Specialist Study Director - Date Dat e 8 T-7098.4 Acute Inhalation Toxicokinctic Study of Perfluoroclancsulfonyl Fluoride (POSF) Final Report Table 1: Control Group Biological Data Exposure animal number 5/1/01 Animal weight Pre exposure Day 0(g) 5/2/01 5/3/01 Animal weight Animal Post exposure weight Post Day 1 (g) exposure Day 2(g) 5/7/01 Animal weight Post exposure Day 6 (g) 5/14/01 Animal T-Test 5/14/01 p-value Necropsy Liver/BW ratio notes weight (g) Predose Liver weights vs day 6 (9) BW (1) Control 1 R01462 Control 2 R01463 Control 3 R01464 avg SD 374 372 369.1 376 391 0.04 14.1 0.036 368 364.5 360.1 373 386 13.1 0.034 392 376 386 401 422 15 0.036 (overnight fast) 378 371 372 383 400 14.1 0.035 12 6 13 15 20 1 0.001 (1) p-va ue for a 2-tailed Students' T-test, assuming equal variance. Predose vs. day 6 body weight. Statistically significant if P<0.05 T-7098.4 Acute Inhalation Toxicokinetic Study o f Perfluoroctanesulfonyl Fluoride (POSF) Final Report Table 2: Toxicokinetic Group Biological Data Sacrifice date Day 0(5-1-01) Inhaled Dose POSF(1) Body Weight day 1 Body Weight day 6 Body Weight day Organ Weight post dose post dose 14 post dose 5/2/2001 (day 1 post dose) 5/2/01 (POSF ppm) Animal # 1000 R01468 5/7/2001 (day 6 post dose) 1000 R01471 Predose Body weight 382 426.4 (mg/kg) 150.79 135.08 (9) 378.2 425 (9) 431 (g) Liver (g) 14.6 15.6 5/14/2001 (day 13 post dose) 1000 R01472 351 164.10 350 359 376 13.6 Avg 386 149.99 384 395 14.6 STD 38 14.53 38 50.91 1 (I) Available dose (Amount inhaled over time) = alpha x VT x f x C x t. For this calculation assume that alpha is 100%, (I.e. = 1) so that all of the inhaled volume of the test atmosphere (1000 ppm POSF) is available for absorption (I.e. the available dose n the exposure to the "lungs"). T-7098.4 Acute Inhalation Toxicokinetic Study o f Perfluoroctanesulfonyl Fluoride (POSF) Final Report Table 3: POSF Toxicokinetic Serum PFOS Data T-7098.1 P O S F Inhalation toxicokinetic Study Date of Exposure : 5/2/01 Exposure: 1000 ppm P O S F /4h rs , 3 male rats Pos t D o se Dose (PO SF ppm Anim al * Day 0 (5-101) Inhaled Dose PO SF (2) Predose Body weight (mg/kg) Serum serum ppm volume (1 ) (P F O S ) Available P O S F In inhaled air (mg) % ol Inhaled P O S F In serum at necropsy (ml) (ug/m i) P F O S In serum (mg) (mg) (% ol dose In se ru m ) Day 1 Day 6 1000 R01488 1000 R01471 382 150.79 12.22 426.4 135.08 13.92 8.6 0.105 57.600 0.182% 9.3 0.129 57.600 0.225% D ayl 3 Avg STD 1000 R01472 351 164.10 386 149.99 38 14.53 1 1.60 12.58 1.20 5 7.63 0.058 0.097 0.036 57.600 57.600 0.000 0.101% 0.169% 0.063% (1) Given: (32.3 mis serum)/(kg of body weight). Serum volume =(bw in grams/1000) x 32.3 (2) Serum ppm: Analytical work done by Dave Ehresman. 1:100 dilution of PFOS in serum measured by HPLC/MS. (3) PFOS in serum (mg): [(Serum voIume)(Serum PFOS)] /I000 (4) POSF in inhaled air (mg): (2.0 ml/breath) x (120 breaths/min) x (1 mg/ 1000ml) x (240 min) = 57.6 mg POSF in inhaled air (5) % inhaled POSF in serum: [(PFOS in serum)/(POSF in inhaled air)]* 100 T-7098.4 Acute Inhalation Toxicokinetic Study o f PerfluooctancsulfonyI Fluoride (POSF) Final Report 13 Table 4: POSF Toxicokinetic Serum Data, dav one post-dose, based on urine std curve. Serum from the rat sacrificed day one post dose (Animal #R01468) was initially extracted and quantified by LC/MS/MS based on a PFOS standard curve extracted from urine collected Irom the limit toxicity study in rats (T-7098.3). This was done to get an estimate of serum PFOS levels on day one post-dose assuming an equal extraction of urine PFOS std curve and serum PFOS. Serum from this animal was later extracted and quantified against a standard curve extracted for serum, and therefore this data was not used in any calculations, but it is included here for completeness. Serum PFOS from day 1 post dose toxicokinetic group animal based on urinary PFOS standard curve. PFOS in serum (ppb) using Standard cuive equation toim combination of two urine extracted PFOS standard curves. Dilution lactor R01468 d1 PD R01468 d1 PD Average SD Diluted Y(AUC) X * ppb of PFOS in serum = (Y61687)/564.53 serum PFOS (ppm) 3827145 6670 6.67 3608110 6282 6.28 3717628 6476 6.48 154881 274 0.27 PFOS in undiluted serum (ppm) 4 26.68 4 25.13 2590 1.10 (I) Assuming a I:I relationship between POSF and PFOS. 100% inhaled dose of POSF could be PFOS in urine and serum. 13 T -7098-4 Acute Inhalation Toxicokinetic Study of Pwfluoroctancsulfonyl Fluoride (POSF) Final Report AUC (Ml) 700000 600000 500000 400000 300000 200000 100000 Figure V. extracted urine combined standard curve for PFOS 400 500 ppb PFOS in urine Series t -- I ineai (Senesi) 900 T-7098.4 Acute Inhalation Toxicokinetic Study of Perfluooctanesulfonyl Fluoride (POSF) Final Report Figure 2: Extracted PFOS standard curve in serum Quart B iim sci * Itn m sc i - 1M. siti (l) to c k e l 1 , V iew All) Brockcl 1 r y fG a to ^ i n n 'S # |E ^ tit^ C ifib i& b tv f S j g i i Pfos_AS_JB03 j i Z j j i PIoj_AS_JB04 .7 3 A V P)os_AS_JB05 P)qj_AS_JBOG t g 5 W Plo_AS_JB07 ' Pio AS JBOB S ^ S a m p l W 5 ng/m L Std 10 ng/m L Std 25 ng/m L Sld 50 ng/m L Std lOOnq/m L Std 250 ng/m L Std IOE & iS se sS ftolSTD jgBI p S 'p c in d K - ii.C lc l te a i Mfcr'7''* 1 |f;|A r n fi m tF s | pf|A m pu ntg S 34923 313701 5 000j 4.896 36058 222760 10.000= 14.669 76487 368159 25.000| 23.748 85130 231311 50 000i 57.291 181310 350304 100 000! 91.527 424330 414210 250.000' 253.642 UUnifi MB S ^*r||'^^llpnS^h)fRf^|ii,. 6 fEcxXcLliuudFue-HV^i,* .07Response Low 4 10 ____ 4$.63 Excluded 414^E S 501! 1 4 .5 8 !............ 8 47' 1.46! Responto High 15 15 - .-ZV'; T-7098.4 Acute Inhalation Toxicokinetic Study o f PerfluoroctanesuJfonyl Fluoride (POSF) Final Report Appendix 1 Histopathologv Results . Necropsy 5/3/01 All rats lungs have generalized mild to moderate congestion of alveoli with occasional extravasation of blood around arterioles and bronchioles. This is possibly an agonal change associated with C 02 euthanasia. Some lungs were not well insufflated. Animal # Control R01462: No significant changes in lung and bladder. Toxicokinetic R01468: No significant changes in lung and bladder. Toxicokinetic R01471: One macroscopic focus of atelectasis (collapsed alveoli) is present in one lung lobe. The change was chronic in duration. No significant change is present is bladder. Toxicokinetic R01472: No significant changes in lung and bladder. T-7098.4 Acute Inhalation Toxicokinetic Study o f Perfluooctanesulfonyl Fluoride (POSF) Final Report 18 Appendix 2 Serum PFOS To: Andrew Seacat/US-Corporate/3M/US@3M-Corporate Tommie Tumer/US-Corporate/3M/US cc: John L. Butenhoff/US-Corporate/3M/US@3M-Corporate Paul Lieder/US-Corporate/3M/US@3M-Corporate Sue Tanaka/US-Corporate/3M/US@3M-Corporate Kathy Thompson/US-Corporate/3M/US Deanna J. Luebker/US-Corporate/3M/US@3M-Corporate Subject: PFOS Levels from rats exposed to POSF via inhalation T-7098.3 Andrew and Tommie: The 1 to 100 estimate for the serum levels of PFOS was right on target. The following results are based on the rat serum being diluted 1:100 prior to extraction. The extraction was carried out at pH 3.0 using a single shake out in Ethyl Acetate. The EA was transferred to a clean polypropylene tube and blown to dryness using nitrogen gas. The resultant residue was re-dissolved in 100 uL of 25% acetonitrile and 75% 10 mm ammonium acetate. Of this solution 20 uL were injected onto a Betas'll C18 reversed phase HPLC column (50 x 2 mm, 5 micron particle size). The flow rate was maintained at 0.5 ml/min. A gradient flow of acetonitrile was used to elute the compounds of interest starting with 15% acetonitrile and ending with 65% acetonitrile over 5.5 minutes. Return to initial operational parameters and column equilibration requires and additional 2.5 minutes for a run time cycle of 8.0 minutes per run. The MS was operated in the Electro Spray negative ionization mode with a constant 3.0 kV potential applied to the source. The method used was an MS/MS method involving the transition of the following ions: Compound Q-2 Offset Voltage Base Peak MS/MS Ion Internal Standard (PFHS) 60 volts 299 amu 30 amu 60 volts PFOS 499 amu 80 amu The capillary heater was held at a constant 300 degrees C. All quantitative calculations were based on the MS/MS ion rations between the compound of interest (PFOS) and the internal standard (PFHS). The standard curve ranged from 5 ng/mL (ppb) to 250 ng/mL (ppb). A quadratic curve fit was used with the standard value weighted 1 / X . This curve fit produced an R squared value of 0.9935. i&&L- * - - . T7Q98.4- Acute Inhalation Toxicokinetic .Study of Perfluooctanesuifonyl Fluoride (POSF) Final'Report 19 .. fi k * - The first tube of two available lor control animal number t R01464 was diluted and . run with the initial extractions. This tube (1:100 dilution) had a PFOS level of 2.3 ppm. Repeat of this tube dilution produced comparable results (2.1 ppm). The second tube labeled 1R01464 was diluted 1:100 and extracted. The second tube on . this control animal had a result of less than the lowest standard (5.0 ppb on the ^diluted serum). Only one serum tube was available for animal 1R01465, unfortunately this tube was labeled 1R0465 with the correct date and T # recorded. I diluted and extracted this tube and recorded results under the animal number 1R01465. The sample for 1R01468 was noted as hemolyzed (2+ hemolysis). The initial result of 1R01472 appeared to be significantly lower than 1R01468 and 1R01471. This ~ sample was also re-extracted and the analysis repeated. No significant difference . was noted on re-extraction. Results: Rat# 1R01462 1R01463 1R01464 Control yes yes yes PFOS Level <5.0 <5.0 <5.0 ppb (on diluted serum) n 1R01465 1R01466 1R01467 1RD1468 1R01471 1R01472 no no no no no no 5.1 ppm .... 4.8 ppm 3.8 ppm 8.6 ppm 9.3 ppm 5.0 ppm (5.1 ppm on duplicate) dje David J. Ehresman 3M Corporate Toxicology "Bldg! 236-1B -22' ' Phone 651-733-5070 FAX 651-737-4754 djehresman @ mmm.com 19 SU BJECT PAGE A PPEN D IX E - D EV IA TIO N S FRO M TH E PR O TO C O L AND TH E STA N D A R D O PER A TIN G PR O C ED U R ES OF TH E TESTIN G FA CILITY E -l to E-3 A PPEN D IX F - C ER TIFIC A TE OF A N A LY SIS F -l to F-3 A PPEN D IX G - A N A LY TICA L AND B IO A N A LY TIC A L REPORT G -I to G-153 A PPEN D IX H - T E M PE R A T U R E A N D R E LA TIV E H U M ID ITY REPORT H -l A PPEN D IX I - PO SITIV E C O N TR O L D A TA 1-1 to 1-4 A PPEN D IX J - H ISTO PA TH O LO G Y R EPO R T J-l toJ-105 A PPEN ID X K - H EM A TO LO G Y A N D C L IN IC A L CH EM ISTRY REPORTS K -l to K -150 A PPEN D IX L - STA TEM EN T OF TH E STU D Y D IRECTO R L -l A PPEN ID IX M - Q U A LITY A SSU R A N C E STA TEM EN T M -l to M -2