Document VJvYGBv76oNm7am6xE6njBYmw

ADVANCED BIOANALYTICAL SERVICES, INC. T 'prA r-T i'.n-pr^ 95MYHW01.3M : AUS 3 ' m 1 UC Ion Spray LC/MS Determination of Perfluoro Analytical Standards Provided by 3M Medical Department Prepared for 3M Medical Department, Toxicology Services, St. Paul, MN 55133-3220 Henry Y. Wu, Ph.D. Jack D. Henion, Ph.D. Advanced BioAnalytical Services, Inc. Date: August 30, 1995 005902 ADVANCED BIDANALYTICAL SERVICES, INC. TABLE OF CONTENT 95MYHW01.3M Page SIGNATURE PAGE ................................................................................................................. i SUMMARY ............................................................................................................................... ii 1. INTRODUCTION ............................................................................................................... 1.1 Study O bjectives.................................................................................................... 1.2 Chemical Structures, Names or 3M Identifications, and Molecular W eights........ 1 1 1 2. MATERIALS AND METHODS .......................................................................................... 2.1 Chemicals and M aterials........................................................................................ 2.2 LC/MS Instrumentation .......................................................................................... 2.3 Preparation of Stock and Test Solutions ................................................................ 2 2 3 5 3. RESULTS AND DISCUSSION .................................................................................. 3.1 T-6295 .................................................................................................................... 3.2 T-6294 .................................................................................................................... 3.3 T-6293 .................................................................................................................... 3.4 Fose Amide ............................................................................................................. 3.5 Perfluorooctanamide ............................................................................................... 5 6 6 7 7 8 4. CONCLUSION .................................................................................................................... 8 5. FIGURES .............................................................................................................................. Figure 1. Total Ion Chromatogram (TIC) for the Injection of 1 |iL of DMSO ............ Figure 2. TIC for the Injections of 1 |iL of Top, Middle and Bottom Day Zero Solution of T-6295 ........................................................................................... Figure 3. TIC for the Injections of 1 |lL of Top, Middle and Bottom Day Seven Solution of T-6295 ........................................................................................... Figure 4. Extracted Ion Chromatograms for the Injection of 1 fiL of Day Zero Top Solution of T-6295 ........................................................................................... Figure 5. Full-Scan Mass Spectrum of Peak 1 in Figure 4 9 9 10 11 12 13 005903 ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 6. Full-Scan Mass Spectrum of Peak 2 in Figure 4 Figure 7. Full-Scan Mass Spectrum of Peak 3 in Figure 4 Figure 8. Full-Scan Mass Spectrum of Peak 4 in Figure 4 Figure 9. Full-Scan Mass Spectra of Peaks 5a and 5b in Figure 4 Figure 10. Full-Scan Mass Spectrum of Peak 6 in Figure 4 ........................................ 18 Figure 11. Full-Scan Mass Spectrum of Peak 7 in Figure 4 ........................................ 19 Figure 12. TIC for the Injections of 1 pL of Top, Middle and Bottom Day Zero Solution of T-6294 ........................................................................................... 20 Figure 13. TIC for the Injections of 1 jiL of Top, Middle and Bottom Day Seven Solution of T-6294 ............................................................................................. 21 Figure 14. Extracted Ion Chromatograms for the Injection of 1 pL of Day Seven Top Solution of T-6294 ................................................................................... 22 Figure 15. Full-Scan Mass Spectrum of Peak 1 in Figure 1 4 ........................................ 23 Figure 16. Full-Scan Mass Spectrum of Peak 2 in Figure 1 4 ........................................ 24 Figure 17. Full-Scan Mass Spectrum of Peak 3 in Figure 1 4 ........................................ 25 Figure 18. Full-Scan Mass Spectrum of Peak 4 in Figure 1 4 ........................................ 26 Figure 19. Full-Scan Mass Spectrum of Peak 5 in Figure 1 4 ........................................ 27 Figure 20. Total Ion Chromatogram (TIC) for the Injection of 1 pL of Water ............ 28 Figure 21. TIC for the Injections of 1 pL of Top, Middle and Bottom Day Zero Solution of T-6293 ........................................................................................... 29 Figure 22. TIC for the Injections of 1 pL of Top, Middle and Bottom Day Seven Solution of T-6293 ........................................................................................... 30 Figure 23. Extracted Ion Chromatograms for the Injection of 1 pL of Day Seven Bottom Solution of T-6293 ................................................................................ 31 Figure 24. Full-Scan Mass Spectrum of Peak 1 in Figure 2 3 ........................................ 32 Figure 25. Full-Scan Mass Spectrum of Peak 2 in Figure 2 3 ........................................ 33 Figure 26. Full-Scan Mass Spectrum of Peak 3 in Figure 2 3 ........................................ 34 Figure 27. TIC for the Injections of 1 pL of Top, Middle and Bottom Day Zero Solution of Fose Amide ................................................................................... 35 Figure 28. TIC for the Injections of 1 pL of Top, Middle and Bottom Day Seven Solution of Fose Amide ................................................................................... 36 Figure 29. Extracted Ion Chromatograms for the Injection of 1 pL of Day Zero Top Solution of Fose Amide ................................................................................... 37 Figure 30. Full-Scan Mass Spectra of Peaks la and lb in Figure 2 9 .......................... 38 1 1 1 17 00S904 ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 31. Full-Scan Mass Spectrum of Peak 2 in Figure 2 9 ...................................... 39 Figure 32. Full-Scan Mass Spectrum of Peak 3 in Figure 2 9 ...................................... 40 Figure 33. Full-Scan Mass Spectrum of Peak 4 in Figure 2 9 ...................................... 41 Figure 34. Full-Scan Mass Spectrum of Peak 5 in Figure 2 9 ...................................... 42 Figure 35. Full-Scan Mass Spectrum of Peak 6 in Figure 2 9 ................... 43 Figure 36. Full-Scan Mass Spectrum of Peak 7 in Figure 2 9 ...................................... 44 Figure 37. TIC for the Injections of 1 (iL of Top, Middle and Bottom Day Zero Solution of Perfluorooctanamide..................................................................... 45 Figure 38. TIC for the Injections of 1 |iL of Top, Middle and Bottom Day Seven Solution of Perfluorooctanamide..................................................................... 46 Figure 39. Extracted Ion Chromatograms for the Injection of 1 \x L of Day Zero Top Solution of Perfluorooctanamide..................................................................... 47 Figure 40. Full-Scan Mass Spectrum of Peak 1 in Figure 3 9 ...................................... 48 Figure 41. Full-Scan Mass Spectrum of Peak 2 in Figure 3 9 ...................................... 49 Figure 42. Full-Scan Mass Spectrum of Peak 3 in Figure 3 9 ...................................... 50 Figure 43. Full-Scan Mass Spectrum of Peak 4 in Figure 3 9 ...................................... 51 Figure 44. Full-Scan Mass Spectrum of Peak 5 in Figure 3 9 ...................................... 52 Figure 45. Full-Scan Mass Spectrum of Peak 6 in Figure 3 9 ...................................... 53 Figure 46. Full-Scan Mass Spectrum of Peak 6 in Figure 3 9 ...................................... 54 i 005905 ADVANCED BIOANALYTICAL SERVICES, INC. ADVANCED BIOANALYTICAL SERVICES 95MYHW01.3M SIGNATURE PAGE Title: Ion Spray LC/MS Determination of Perfluoro Analytical Standards Provided by 3M Medical Department Reported by: Senior Research Scientist 8W ?Jr Date Reviewed by: tils / "Mary. Quality Assurance Manager Authorized for Release by: (^Jacl^H enion, Ph.D. ABS Laboratory Director 'Date U S 9 0 6 ADVANCED BIOANALYTICAL SERVICES, INC. SUMMARY 95MYHW01.3M Five perfluoro compounds including T-6293, T-6294, T-6295, fose amide and perfluorooctanamide were analyzed by full-scan ion spray LC/MS for composition identification and ambient temperature stability in DMSO and water (only for T-6293). The on-line LC/MS determination of the listed test articles was carried out under unit mass resolution conditions following mass axis calibration using PPG reference compounds. Chromatography was performed on a 2 x 100 mm DeltabondTM C8 column using linear gradient elution with acetonitrilewater from 5:95 to 95:5 in 15 minutes. The mass spectrometer was operated in the negative ion mode. The LC/MS experiments showed that all five perfluoro compounds contain some impurities. In addition, all five perfluoro compounds are stable in DMSO and water for at least a week at ambient temperature at a concentration of 10 mM. No apparent degradation was observed by LC/MS analyses of these samples. Attempts to obtain molecular ion information for compounds T-6292 and butyl fose by ion spray and atmospheric pressure chemical ionization (APCI) were unsuccessful. However, poor response from both ionization modes did not provide sufficient sensitivity for the LC/MS analysis of these two compounds. m 005907 ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M 1. INTRODUCTION Seven perfluoro compounds including T-6292, T-6293, T-6294, T-6295, fose amide, butyl fose and perfluorooctanamide were the target molecules for qualitative composition and stability determination. Among them, five perfluoro compounds including T-6293, T-6294, T-6295, fose amide and perfluorooctanamide were analyzed by full-scan ion spray LC/MS for composition identification and ambient temperature stability in DMSO and water (only for T-6293). The on-line LC/MS determination of the listed test articles was carried out under unit mass resolution conditions following mass axis calibration using PPG reference compounds. Chromatography was performed on a 2 x 100 mm DeltabondTM C8 column using linear gradient elution with acetonitrile-water from 5:95 to 95:5 in 15 minutes. The mass spectrometer was operated in the negative ion mode. The LC/MS experiments showed that all five perfluoro compounds contain some impurities. In addition, all five perfluoro compounds are stable in DMSO and water for at least a week at ambient temperature at a concentration of 10 mM. No apparent degradation was observed by LC/MS analyses of these samples. Attempts to obtain molecular ion information for compounds T-6292 and butyl fose by ion spray and atmospheric pressure chemical ionization (APCI) were unsuccessful. Poor response from both positive and negative ionization modes did not provide sufficient sensitivity for the LC/MS analysis of these two compounds. 1.1 Study Objectives The objectives of this study were: * to qualitatively determine the composition of each perfluoro compound. * to qualitatively determine the stability of each compound in DMSO or water (for T-6293 only) as a stock solution at 10 mM concentration. 1.2 Chemical Structures, Names or 3M Identifications, and Molecular Weights O C8Fn -----sII--- O K + o T-6295 MW = 538.22 anion m/z = 499 005908 ADVANCED BIOANALYTICAL SERVICES, INC. CgFi? Co8MF 7 C8SrF17 Co8F"17 C7F,5 o CH,2C^H3 / SN H O o CH2CH3 s-- nCT II ^ ch2ch2o - o o c h 2c h 3 - s - N oII ch2ch2oh o II - s - NH, o o II NH, o II P---- O' n h 4+ o n h 4+ Co8rF17 O c h 2c h 2c h 2 ch3 n : c h 2c h 2oh o 95MYHW01.3M T-6294 MW = 527.20 anion m/z = 526 T-6293 MW = 685.29 monoanion m/z = 650 T-6292 MW = 571.25 fose amide MW = 499.14 anion m/z = 498 Perfluorooctan amide MW = 413.09 anion m/z = 412 butyl fose MW = 599.31 2. MATERIALS AND METHODS 2.1 Chemicals and Materials All seven perfluoro compounds were provided by 3M Medical Department, Toxicology Services, St. Paul, MN 55133-3220. Methyl sulfoxide (DMSO, Cat #27,043-1) was obtained from SigmaAldrich Chemicals, Milwaukee, WI 53233. Acetonitrile (Cat #015-4) was obtained from 2 005909 ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M Baxter/Scientific Products, McGaw Park, IL 60085. Ammonium Acetate (Cat #JT0596-0I) was purchased from J.T. Baker, Danvers, MA 01923. High purity (NANOpure) water was made by Bamstead Model D7331 Ultrapure Water System, Dubuque, IA 52001. All materials and equipment may be substituted by equivalent materials and equipment. Fisher Vortex Genie 2TM, Cat #12-812, Fisher Scientific, Springfield, NJ 07081 Microbalance, Model MT5, Mettler-Toledo, Inc., Hightstown, NJ 08520 Siliconized Graduated Tubes, 2 mL, Cat #M-720, Phnix Research Products, Hayward, CA 94545 Pipettes, Cat #P-1000, P-200, P-100, P-20, Rainin Instrument Co., Woburn, MA 01888 Siliconized Green Slick Tips, 1 mL, Cat #TGS-415BRS, Phnix Research Products, Hayward, CA 94545 Siliconized Clear Tips, 250 p.L, Cat #T-051 BRCS, Phnix Research Products, Hayward, CA 94545 Polypropylene autosampler vials (siliconized in house), Cat #500 102, Sun Brokers, Inc., Wilmington, NC 28402 Snap Cap, 11 mm, Cat #69158-P, Scientific Resources Inc., Eatontown, NJ 07724 Solvent Filtration Apparatus, Cat #32-00610, Krackeler Scientific, Albany, NY 12201 Vacuum Pump, Cat #54-907, VWR Scientific, Rochester, NY 14603 0.45 pm Nylon Titan Membrane Filters, Cat #74547-NN, Scientific Resources Inc., Eatontown, NJ 07724 Liquid Nitrogen, Cat #SS180N-NIT4052, Empire Airgas, Elmira, NY 14901 U.H.P. Nitrogen, Empire Airgas, Elmira, NY 14901 U.H.P. Argon, Empire Airgas, Elmira, NY 14901 Fused-silica, 100 p. i.d. x 170 p. o.d., Polymicro Technologies, Phoenix, AZ 85023 PEEK tubing, 0.007" i.d. x 1/16" o.d., Cat. #1536, Upchurch Scientific, Inc., Oak Harbor, WA 98277 2.2 LC/MS Instrumentation The liquid chromatography part of the system included a Hewlett-Packard Model 1090L pump, Hewlett-Packard, Avondale, PA 19311. DeltabondTM C8 columns, 5 pm, 2 x 100 mm, and DeltabondTM C8 guard cartridges (5 pm, 2 x 1 0 mm) were obtained from Keystone Scientific, Inc., 3 005910 ADVANCED BIOANALYTICAL SERVICES. INC. 95MYHW01.3M Bellefonte, PA 16823. The mass spectrometer was a PE-SCIEX API III+ from PE-SCIEX, Thornhill, Ontario. HPLC Conditions: Flow Rate Column Temperature Injection Volume Column Guard Cartridge Pressure Eluents t (min) 0 15 200 pL/min Ambient 1 pL 2 x 100 mm DeltabondTM C8 2 x 1 0 mm DeltabondTM C8 70 bar %acetonitrile 5 95 %water 95 5 This program was used for the analyses of T-6294, T-6295, fose amide and perfluorooctanamide. For the analysis of T-6293 samples, water was replaced by 10 mM ammonium acetate with the same gradient program. Mass Spectrometer Conditions: Ion Spray Voltage Orifice Voltage Declustering Potential Multiplier Voltage Curtain Gas (U.H.P. Nebulizer Gas (Nz) Interface Heater Turbospray Temperature Turbospray Gas Flow Dwell Time Mass Scan Range Scan Rate -4200 V -90 V (-70 V for T-6293) -60 V (-40 V for T-6293) 4000 V 1.4 L/min 50-70 psi 60 C 500 C 4.0 L/min 1 msec 100-900 Da 1.69 sec/scan 4 005911 ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M Count Controller (CC) 10 Mass calibration of Q1 in the range of 100-900 was performed daily by the infusion of a 10-4M PPG (polypropylene glycols, average MW 425, 1000 and 2000) calibration solution (PE-SCIEX API IH+ operator's manual) at a flow of 10 jiL/min. Calibration was performed on m/z 59.0, 325.3, 500.38 and 906.7. The mass spectrometric parameters, MS/MS peak widths and sensitivity are optimized daily. Peak widths are approximately 0.7 amu at half-height in the single MS mode. 2.3 Preparation of Stock and Test Solutions Stock Solutions: Accurately weigh approximately 5 mg of one of the perfluoro compounds (assuming purity 100%) on a micro balance in a siliconized graduated tube (2 mL). Dissolve the solid sample in an appropriate amount of DMSO or water (for T-6293 only) to yield a 10 mM solution. Store the solution at ambient temperature. Test Solutions: Carefully take 10 |nL of the 10 mM stock solution from the top, middle and bottom parts of the solution, and dilute them with DMSO or water (for T-6293 only) by 10- to 100-fold to give test solutions with concentrations of 100-1000 pM. Vortex-mix the diluted solutions, and analyze them immediately after preparation. The test solutions with same concentrations for the analytes were prepared on day zero and day seven in the same way from the same stock solution. The stock solutions were stored at ambient temperature on the lab bench without disturbance. 3. RESULTS AND DISCUSSION As requested by 3M, the polypropylene tubes and vials used in this work were siliconized. The pipette tips used in this work were also siliconized. The LC/MS conditions for the analyses of different parts of a solution on both day zero and day seven were the same. To avoid cross contamination, solvent blanks (DMSO or water) were injected before the injection of a different type of test solution. No LC/MS data were obtained for compounds T-6292 and butyl fose, because the molecular ion signal was too weak to be used to perform LC/MS experiments. The on-line LC/MS determination of the five compounds tested was carried out under unit mass 5 005912 ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M resolution conditions following mass axis calibration using PPG reference compounds. To achieve sufficiently fast full-scan acquisition scan rates for these relatively higher molecular weight compounds, a compromise of 0.5 Da step size acquisition was used in these experiments. These conditions produce plots of corresponding mass spectra from the proprietary PE SCIEX software that do not include the isotopic peaks. Mass spectral plots of individual test article mass spectra from these acquisitions show reliable molecular weight measurements. In particular, differentiation of molecular weights differing by only one dalton, e.g. the sulfonamide vs. the sulfonic acid at m/z 498 and 499, may be reliably determined under these conditions. 3.1 T-6295 A typical total ion chromatogram (TIC) from the injection of 1 pL of DMSO is shown in Figure 1. The TIC from the injections of top, middle and bottom parts of the T-6295 solution on day zero and day seven were summarized in Figures 2-3. Extracted ion chromatograms from the injection of 1 pL of day zero test solution (100 pM) diluted from the top part of the stock solution are shown in Figure 4. Full-scan single MS spectra of the components detected from Figure 4 are shown in Figures 5-11. The identical mass spectra in Figure 9 for peaks 5a and 5b suggest that these two peaks are likely from two structural isomers. By carefully examining the TIC from the injections of solvent blank and different parts of the T-6295 solutions on both day zero and day seven, there is no apparent difference among the three parts of the solution and there is no apparent change in the composition of the stock solution at ambient temperature for a week. Although trace impurities at m/z 299, 349, 399, 449, 549 and 607 are detected in T-6295, the major species present is the m/z 499 anion. 3.2 T-6294 The TIC from the injections of top, middle and bottom parts of the T-6294 solution on day zero and day seven were summarized in Figures 12-13. Extracted ion chromatograms from the injection of 1 pL of day seven test solution (100 pM) diluted from the top part of the stock solution are presented in Figure 14. Full-scan single MS spectra of the components detected from Figure 14 are shown in Figures 15-19. By carefully examining the TIC from the injections of solvent blank and different parts of the T-6294 solutions, it appears that the bottom part of the solution contains fewer impurities. In addition, ions at m/z 297.5, 311.5, 325.5 and 339.5 showed higher relative responses (compared to the ion at m/z 526 for [M-H]'of T-6294) for the day seven 6 005913 ADVANCED BIDANALYTICAL SERVICES, INC. 95MYHW01.3M solutions than those obtained for the day zero solutions (See Figures 12 and 13 in the regions of 5.6-8.5 min.). Again, the m/z 526 anion is the major species in the sample. 3.3 T-6293 T-6293 was not soluble in DMSO. Therefore, water was used to prepare the stock and test solutions for T-6293. A typical TIC from the injection of 1 pL of water is shown in Figure 20. Total ion chromatograms from the injections of top, middle and bottom parts of the T-6293 solution on day zero and day seven are summarized in Figures 21-22. Extracted ion chromatograms from the injection of 1 |iL of day seven test solution (1 mM) diluted from the bottom part of the stock solution are presented in Figure 23. Full-scan single MS spectra of the components detected from Figure 23 are shown in Figures 24-26. Components at m/z 650, 630 and 369 were detected. Based on infusion experiments, the ion at m/z 526 in Figure 25 is a fragment of m/z 650. By carefully examining the TIC from the injections of solvent blank and different parts of the T-6293 solutions on both day zero and day seven, there is no apparent difference among the three parts of the solution and there is no apparent change in the composition of the stock solution at ambient temperature for a week, although the chemical background for the day seven solutions seems to be noisier possibly due to lower sensitivity of the mass spectrometer. For unknown reasons, the chemical background for the day zero top and middle solutions are somewhat higher than that for the day zero bottom solution (see Figure 21). 3.4 Fose Amide The TIC from the injections of top, middle and bottom parts of the solution of fose amide on day zero and day seven were summaried in Figures 27-28. Extracted ion chromatograms from the injection of 1 p.L of day zero test solution (100 |iM) diluted from the top part of the stock solution are presented in Figure 29. Full-scan single MS spectra of the components detected in Figure 29 are shown in Figures 30-36. Components at m/z 606, 499, 498, 398, 339.5, 325.5, and 311.5 were detected. Peaks marked with an X are from the chemical background of the LC eluent (see Figures 34 and 36). Interestingly, the top portion of the day zero solution showed much higher relative response for the ions at m/z 499, 339.5, 325.5 and 311.5 than those obtained from the middle and bottom portions of the same day solution. On the contrary, the day seven solution showed the opposite trend for the responses of those ions. The ion at m/z 499 appeared at two different retention times (see peak la and lb in Figure 29). Peak la is possibly from an impurity, while peak lb is the isotopic form of the major component at m/z 498 (see Figures 30 and 35). By 7 005914 ADVANCED BIDANALYTICAL SERVICES, INC. 95MYHW01.3M carefully examining the TIC from the injections of solvent blank and different parts of the solutions of fose amide on both day zero and day seven, there is no significant difference between the three parts of the solution and there is no apparent change in the composition of the stock solution at ambient temperature for a week. 3.5 Perfluorooctanamide The TIC from the injections of top, middle and bottom parts of the solution of perfluorooctanamide on day zero and day seven are summarized in Figures 37-38. Extracted ion chromatograms from the injection of 1 pL of day zero test solution (1 mM) diluted from the top part of the stock solution are presented in Figure 39. Full-scan single MS spectra of the components detected in Figure 39 are shown in Figures 40-46. Major components at m/z 412, 392, 369, 339.5, 325.5, 311.5 and 297 were detected. Peaks marked with an X are from the chemical background of the LC eluent (see Figure 46). Again, the top portion of the day zero solution showed much higher relative response for the ions at m/z 339.5, 325.5 and 311.5 than those obtained from the middle and bottom portions of the same day solution. The relative responses for the ions at m/z 339.5, 325.5 and 311.5 for the day seven solution were significantly lower than those obtained for the day zero solution. It is clear from Figure 39 that this sample has several components present in addition to the test article of interest. 4. CONCLUSION Five perfluoro compounds were analyzed by ion spray LC/MS. In general, all five compounds are stable in DMSO or water (for T-6293 only). No significant degradation products were detected for any of the compounds analyzed. However, it appears that the low molecular weight impurities are not uniformly distributed in some solutions. 8 005915 ADVANCED BIDANALYTICAL SERVICES, INC. 5. FIGURES 95MYHW01.3M Figure 1. Total Ion Chromatogram (TIC) for the Injection of 1 |iL of DMSO TIC of all masses, 100 to 900 DMSO LC/MS-3- 7/5/95 -10:45 AM 1 uL of DMSO.deltabond C8.ACN/H20 gradient(5% to 95% in 15 min) 55,000,000 Scan/Time (min) 9 005916 ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 2. TIC for the Injections of I (iL of Top, Middle and Bottom Day Zero Solution of T-6295 Relative Intensity (%) Relative Intensity (%) Relative Intensity (%) Scan/Time (min) 10 005917 Relative Intensity (%) Relative Intensity (%) ADVANCED B IOANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 3. TIC for the Injections of 1 jiL of Top, Middle and Bottom Day Seven Solution of T-6295 11 00591$ Relative Intensity (%) ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 4. Extracted Ion Chromatograms for the Injection of 1 jiL of Day Zero Top Solution of T-6295 T-6295 LC/MS-1 - 7/5/95 -1 1 :11 AM 1 uL(100 uM/top)T-6295,deltabond C8,ACN/H20(5% to 95% in 15 min) 100/900 TIC 5,400,000 299/299 u 349/349 i 399/399 >. w0c) 449/449 > 1] &t 499/499 A A 5b 20,000 60,000 130,000 100,000 2 , 100,000 549/549 607/607 1 0.0 2.5 1 90 30,000 6 l, 7 30,000 | 5.0 7.5 10.0 12.5 15.0 178 267 356 444 533 Time (min)/Scan 12 005919 ADVANCED BIOANALYTICAL SERVICES, INC. Figure 5. Full-Scan Mass Spectrum of Peak 1 in Figure 4 T-6295 LC/MS-1/Scans 40-42 -Profile Q1 SCAN Scans 40-42 minus 1-35 & 45-700 Time=1.13 min T-6295 LC/MS-1 - 7/5/95 -1 1 :11 AM 1 uL(100 uM/top)T-6295,deltabond C8,ACN/H20(5% to 95% in 15 min) 2 peaks 95MYHW01.3M 9970 13 005920 ADVANCED BIOANALYTICAL SERVICES, INC. Figure 6. Full-Scan Mass Spectrum of Peak 2 in Figure 4 T-6295 LC/MS-1/Scans 57-63 -Profile Q1 SCAN Scans 57-63 minus 1-54 & 72-700 Time=1.66 min T-6295 LC/MS-1 - 7/5/95 -1 1 :11 AM 1 uL(100 uM/top)T-6295,deltabond C8,ACN/H20(5% to 95% in 15 min) 1 peak 100n 349.0 95MYHW01.3M 22,857 75- ccn 50 (>1) jCj(Co0 25 1--r . 200 400 600 800 m/z 14 005921 ADVANCED BIO A N ALYTIC AL S E R V IC E S , INC. Figure 7. Full-Scan Mass Spectrum of Peak 3 in Figure 4 -Profile Q1SCAN T-6295 LC/MS-1/Scans 143-159 Scans 143-159 minus 1-135 & 178-700 Time=4.23 min T-6295 LC/MS-1 - 7/5/95 -1 1 :11 AM 1 uL(100 uM/top)T-6295,deltabond C8,ACN/H20(5% to 95% in 15 min) 1 peak 95MYHW01.3M 74,118 005922 15 C a t h e r w o o d Road It haca. New York 14850 (607) 2 6 6 - 0 6 6 5 Fax (607) 2 6 6 - 0 7 4 9 ADVANCED BIOANALYTICAL S E R V IC E S , INC. Figure 8. Full-Scan Mass Spectrum of Peak 4 in Figure 4 -Profile Q1 SCAN T-6295 LC/MS-1/Scans 197-205 Scans 197-205 minus 1-178 & 214-700 Time=5.64 min T-6295 LC/MS-1 - 7/5/95 -11:11 AM 1 uL(100 uM/top)T-6295,deltabond C8,ACN/H20(5% to 95% in 15 min) 2 peaks 95MYHW01.3M 63,333 16 00592a - 15 C a t h e r w o o d Road It haca. New York 14850 (607) 2 6 6 - 0 6 6 5 Fax (607) 2 6 6 - 0 7 4 9 ADVANCED B IO A N A LY TIC AS E R V IC E S , INC. Figure 9. Full-Scan Mass Spectra of Peaks 5a and 5b in Figure 4 95MYHW01.3M -Profile Q1SCAN T-6295 LC/MS-1/Scans 220-225 Scans 220-225 minus 1-196 & 285-712 Time=6.25 min T-6295 LC/MS-1 - 7/5/95 -1 1 :11 AM 1 uL(100 uM/top)T-6295,deltabond C8,ACN/H20(5% to 95% in 15 min) 1 peak 1,204,860 m/z -Profile Q1SCAN T-6295 LC/MS-1/Scans 228-236 Scans 228-236 minus 1-180 & 400-700 Time=6.51 min T-6295 LC/MS-1 - 7/5/95 - 11:11 AM 1 uL(100 uM/top)T-6295,deltabond C8,ACN/H20(5% to 95% in 15 min) 005924 15 Cat her wood Road Ithaca. New York 14850 (607) 26 6- 0 6 6 5 Fax (607) 266- 0749 ADVANCED BID ANALYTIC AL S E R V IC E S , INC. Figure 10. Full-Scan Mass Spectrum of Peak 6 in Figure 4 T-6295 LC/MS-1/Scans 243-245 -Profile Q1SCAN Scans 243-245 minus 1-240 & 248-712 Time=6.85 min T-6295 LC/MS-1 - 7/5/95 -1 1 :11 AM 1 uL(100 uM/top)T-6295,deltabond C8,ACN/H20(5% to 95% in 15 min) 4 peaks 95MYHW01.3M 156,683 00S92S " 15 Cat her wood Road Ithaca. New York 14850 (607) 2 6 6- 0665 Fax (607) 266- 0749 ADVANCED BIOANALYTICAL S E R V IC E S , INC. Figure 11. Full-Scan Mass Spectrum of Peak 7 in Figure 4 T-6295 LC/MS-1/Scans 260-262 -Profile Q1SCAN Scans 260-262 minus 1-256 & 268-700 Time=7.33 min T-6295 LC/MS-1 - 7/5/95 -11:11 AM 1 uL(100 uM/top)T-6295,deltabond C8,ACN/H20(5% to 95% in 15 min) 3 peaks 95MYHW01.3M 101,898 005926 15 Cat h er wo o d Road Ithaca. New York 14850 (607) 266- 0665 Fax (607) 266- 0749 ADVANCED WiWBBIH B IO A N A L Y T I C A L fia H H S E R V I C E S , I N C . 95MYHW01.3M Figure 12. TIC for the Injections of 1 |i.L of Top, Middle and Bottom Day Zero Solution of T-6294 Scan/Time (min) > 0 0 5 9 27 15 Cat her wood Road Ithaca, New York 14850 (607) 266- 0665 Fax (607) 266-074S ADVANCED BIOANALYTICAL S E R V IC E S . INC. 95MYHW01.3M Figure 13. TIC for the Injections of 1 fiL of Top, Middle and Bottom Day Seven Solution of T-6294 Relative Intensity (%) Relative Intensity (%) Relative Intensity (%) Scan/TIme (min) 005928 15 Ca t h e r w o o d Road Ithaca. New York 14850 (607) 2 6 6 - 0 6 6 5 Fax (607) 2 6 6- 0 7 4 9 ADVANCED B D A N A L Y T IC A L S E R V IC E S , INC. 95MYHW01.3M Figure 14. Extracted Ion Chromatograms for the Injection of 1 |xL of Day Seven Top Solution of T-6294 T-6294 LC/MS-1 - 7/12/95 -1:08 PM 1 ul of T-6294(100 uM/top),delta C8,ACN/H20{5%ACN to 95%ACN in 15 min) 100/900 3,710,000 311.5/311.5 10O 50 DaC> 0 325.5/325.5 100 50 tr O 339.5/339.5 100 50 QC 0 526/526 1 250,000 190,000 4M 70,000 1,460,000 Time (min)/Scan 005929 22 15 C a t h e r w o o d Road Ithaca. New York 14850 (607) 2 6 6 - 0 6 6 5 Fax (607) 2 6 6- 0749 ADVANCED BIDANALYTICAL S E R V IC E S , INC. Figure 15. Full-Scan Mass Spectrum of Peak 1 in Figure 14 -Profile Q1SCAN T-6294 LC/MS-1/Scans 212-221 Scans 212-221 minus 1-210 & 225-712 Time=6.08 min T-6294 LC/MS-1 - 7/12/95 -1:08 PM 1 ul of T-6294(100 uM/top),delta C8,ACN/H20(5%ACN to 95%ACN in 15 min) 2 peaks 95MYHW01.3M 36,447 23 005930 15 C a t h e r w o o d Road It haca. New York 14850 (607) 2 6 6 - 0 6 6 5 Fax (607) 2 6 6- 0 7 4 9 ADVANCED BIOANALYTICAL S E R V IC E S , INC. Figure 16. Full-Scan Mass Spectrum of Peak 2 in Figure 14 T-6294 LC/MS-1/Scans 230-234 -Profile Q1SCAN Scans 230-234 minus 1-220 & 245-712 Time=6.51 min T-6294 LC/MS-1 - 7/12/95 - 1 :08 PM 1 ul of T-6294(100 uM/top),delta C8,ACN/H20(5%ACN to 95%ACN in 15 min) 3 peaks 311.5 95MYHW01.3M 163,658 24 005931 15 C a t h e r w o o d Road It haca. New York 1485 0 (607) 2 6 6 - 0 6 6 5 Fax (607) 2 6 6 - 0 7 4 9 ADVANCED BIOANALYTICAL S E R V IC E S , INC, Figure 17. Full-Scan Mass Spectrum of Peak 3 in Figure 14 T-6294 LC/MS-1/Scans 243-247 -Profile Q1SCAN Scans 243-247 minus 1-232 & 258-712 Time=6.88 min T-6294 LC/MS-1 - 7/12/95 -1:08 PM 1 ul of T-6294(100 uMAop),delta C8,ACN/H20(5%ACN to 95%ACN in 15 min) 4 peaks 325.5 95MYHW01.3M 158,440 25 0 5 3 . 3 2 15 Cat her wood Road Ithaca, New York 14850 (607) 266- 0665 Fax (607) 266- 0749 ADVANCED B IO A N A LY T IC A L S E R V I C E S , IN C . Figure 18. Full-Scan Mass Spectrum of Peak 4 in Figure 14 -Q1 SCAN T-6294 LC/MS-1/Scans 257-260 Scans 257-260 minus 1-250 & 270-712 Time=7.26 min T-6294 LC/MS-1 - 7/12/95 -1:08 PM 1 ul of T-6294(100 uM/top),delta C8,ACN/H20(5%ACN to 95%ACN in 15 min) 3 peaks 100i 339.5 95MYHW01.3M 59,865 75- Relative Intensity (%) 50183.0 25- 343.0 200 400 600 800 m/z 00; 933 26 15 C a t h e r w o o d Road haca. New York 14850 a ( 607) 2 6 6 - 0 6 6 5 a Fax ( 607) 2 6 6 - 0 7 4 9 ADVANCED B IO A N ALYTIC AL S E R V IC E S , IN C . Figure 19. Full-Scan Mass Spectrum of Peak 5 in Figure 14 -Profile Q1 SCAN T-6294 LC/MS-1/Scans 432-435 Scans 432-435 minus 1-410 & 460-712 Time=12.20 min T-6294 LC/MS-1 - 7/12/95 -1:08 PM 1 ul of T-6294(100 uM/top),delta C8,ACN/H20(5%ACN to 95%ACN in 15 min) 3 peaks 95MYHW01.3M 1,232,500 005934 15 C a t h e r w o o d Road It haca. New York 14850 (607) 2 6 6 - 0 6 6 5 Fax (607) 2 6 6 - 0 7 4 9  ADVANCED W lC T W IH b io a n a l y t ic a l K f l H S E R V IC E S , INC. 95MYHW01.3M Figure 20. Total Ion Chromatogram (TIC) for the Injection of 1 |iL of Water TIC of all masses, 100 to 900 H 2 0 LC/MS-4- 7/13/95 -11:56 AM 1 ul of H20,delta C8.ACN/H20 gradlent(5%ACN to 95%ACN in 15 min) 100i 15,000,000 Relative Intensity (%) 201 301 5.6 8.5 Scan/Time (min) 28 0 0 5 3 3 5 15 Cat herwood Road Ithaca. New York 14850 (607) 266-0665 Fax (607) 266-074S ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 21. TIC for the Injections of 1 |liL of Top, Middle and Bottom Day Zero Solution of T-6293 1001 Top 39,800,000 75' 501 Relative Intensity (%) loot Middle 24,400,000 Relative Intensity (%) Relative Intensity (%) 30,700,000 201 301 5.6 8.5 Scan/Time (min) 29 C 0593S ADVANCED BIDANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 22. TIC for the Injections of 1 pL of Top, Middle and Bottom Day Seven Solution of T-6293 Relative Intensity (%) Relative Intensity (%) Relative Intensity (%) 1 101 201 301 401 501 0.0 2.8 5.6 8.5 11.3 14.1 Scan/Tme (min) 30 0Cr>337 ADVANCED BIDANALYTICAL SERVICES. INC. 95MYHW01.3M Figure 23. Extracted Ion Chromatograms for the Injection of 1 jjJL of Day Seven Bottom Solution of T-6293 T-6293 LC/MS-3- 7/13/95 -1 1 :34 AM 1 ul of T-6293(1 mM in H20/bottom),delta C8.ACN/H20 gradient(5%ACN to 95%ACN in 15 .. 100/900 1001 T IC 1,260,000 4-5 C <D CC 369/369 100 75 50 <D CC 25 0 650/650 10O 75 50 Q> CC 25 O 630/630 10O 75 50 05 CC 25 0.0 1 I1 2.5 90 50,000 630,000 ^VTfnfnrMTnwiAnwl, - 300,000 'I 7" 5.0 7.5 10.0 12.5 15.0 178 267 355 444 532 Time (min)/Scan 31 0CS93S ADVANCED BIOANALYTICAL SERVICES, INC. Figure 24. Full-Scan Mass Spectrum of Peak 1 in Figure 23 95MYHW01.3M T-6293 LC/MS-3/Scans 284-285 -Profile Q1SCAN Scans 284-285 minus 1-278 & 289-712 Time=8.00 min T-6293 LC/MS-3 - 7 /1 3 /9 5 -1 1 :34 AM 1 ul of T-6293(1 mM in H20/bottom),delta C8.ACN/H20 gradient(5%ACN to 95%ACN in 15 .. 1 peak 40,000 Relative Intensity (%) 32 V Q 3 33 ADVANCED BIOANALYTICAL SERVICES, INC. Figure 25. Full-Scan Mass Spectrum of Peak 2 in Figure 23 95MYHW01.3M T-6293 LC/MS-3/Scans 314-323 -Profile Q1 SCAN Scans 314-323 minus 1-284 & 450-712 Time=8.96 min T-6293 LC/MS-3 - 7/13/95 - 1 1 :34 AM 1 ul of T-6293(1 mM in H20/bottom),delta C8.ACN/H20 gradient(5%ACN to 95%ACN in 15 min) 2 peaks 650.0 505,068 33 005340 ADVANCED BIOANALYTICAL SERVICES, INC. Figure 26. Full-Scan Mass Spectrum of Peak 3 in Figure 23 95MYHW01.3M T-6293 LC/MS-3/Scans 412-420 -Profile Q1 SCAN Scans 412-420 minus 1-391 & 440-712 Time=11.72 min T-6293 LC/MS-3 - 7/13/95 - 1 1 :34 AM 1 ul of T-6293(1 mM in H20/bottom),delta C8,ACN/H20 gradient(5%ACN to 95%ACN in 15 min) 2 peaks 630.0 218,889 34 0 0 5 5 4 1 ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 27. TIC for the Injections of 1 |iL of Top, Middle and Bottom Day Zero Solution of Fose Amide Relative Intensity (%) Relative Intensity (%) Relative Intensity (%) Scan/Time (min) 35 605342 ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 28. TIC for the Injections of 1 pJL of Top, Middle and Bottom Day Seven Solution of Fose Amide Relative Intensity (%) Relative Intensity (%) Relative Intensity (%) Scan/Time (min) 36 3 ADVANCED BO ANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 29. Extracted Ion Chromatograms for the Injection of 1 (iL of Day Zero Top Solution of Fose Amide Fose Amide LC/MS-1- 7/5/95 - 4:32 PM 1 uL of Fose Amide(100 uM/top),delta C8,ACN/H20(5% to 95% in 15 min) 100/900 ^ T IC 499/499 311.5/311.5 ] \ 325.5/325.5 1, 339.5/339.5 CE 1 398/398 , 1 498/498 , 1 606/606 , , A ... lb la ... .........- A 2 JS ....., . A , 3 A 4 A 5 II , 6 A 7 4,360,000 410,000 120,000 * 170,000 110,000 50,000 1,850,000 30,000 1_--__--___I_--___--____ 1--__I_--___ ^ 0.0 2.5 5.0 7.5 10.0 12.5 15.0 1 90 178 267 355 444 533 Time (min)/Scan 37 005944 ADVANCED BIOANALYTICAL SERVICES. INC. 95MYHW01.3M Figure 30. Full-Scan Mass Spectra of Peaks la and lb in Figure 29 -Profile Q1SCAN Fose Amide LC/MS-1 /Scans 221-225 Scans 221-225 minus 1-196 & 440-712 Time=6.26 min Fose Amide LC/MS-1 - 7/5/95 - 4:32 PM 1 uL of Fose Amide(100 uM/top),delta C8,ACN/H20(5% to 95% in 15 min) 3 peaks [P e a k la 141,722 i i -Profile Q1SCAN Scans 390-397 minus 1-178 & 405-712 Time=11.07 min Fose Amide LC/MS-1 - 7/5/95 - 4:32 PM 1 uL of Fose Amide(100 uM/top),delta C8,ACN/H20(5% to 95% in 15 min) 38 C05.94S ADVANCED BIOANALYTICAL SERVICES, INC, Figure 31. Full-Scan Mass Spectrum of Peak 2 in Figure 29 Fose Amide LC/MS-1/Scans 236-245 -Profile Q1 SCAN Scans 236-245 minus 1-231 & 250-712 Time=6.76 min Fose Amide LC/MS-1 - 7/5/95 - 4:32 PM 1 uL of Fose Amide(100 uM/top),delta C8,ACN/H20(5% to 95% in 15 min) 2 peaks 95MYHW01.3M 393,298 39 005946 ADVANCED BIDAN ALVTICAL S E R V IC E S . INC. Figure 32. Full-Scan Mass Spectrum of Peak 3 in Figure 29 Fose Amide LC/MS-1 /Scans 250-258 -Profile Q1SCAN Scans 250-258 minus 1-235 & 270-712 Time=7.14 min Fose Amide LC/MS-1 - 7/5/95 - 4:32 PM 1 uL of Fose Amide(100 uM/top),delta C8,ACN/H20(5% to 95% in 15 min) 2 peaks 95MYHW01.3M 349,901 40 005947 ADVANCED BIOANALYTICAL SERVICES. INC. Figure 33. Full-Scan Mass Spectrum of Peak 4 in Figure 29 Fose Amide LC/MS-1/Scans 262-271 -Profile Q1 SCAN Scans 262-271 minus 1-250 & 277-712 Time=7.49 min Fose Amide LC/MS-1 - 7/5/95 - 4:32 PM 1 uL of Fose Amide(100 uM/lop),delta C8,ACN/H20(5% to 95% in 15 min) 2 peaks 95MYHW01.3M 77,834 41 0 0 5 9 4 s ADVANCED BIOANALYTICAL SERVICES, INC. Figure 34. Fuli-Scan Mass Spectrum of Peak 5 in Figure 29 Fose Amide LC/MS-1 /Scans 343-344 -Profile Q1SCAN Scans 343-344 minus 1-338 & 355-712 Time=9.66 min Fose Amide LC/MS-1- 7/5/95 - 4:32 PM 1 uL of Fose Amide(100 uM/top),delta C8,ACN/H20(5% to 95% in 15 min) 2 peaks 95MYHW01.3M 45,000 42 005949 ADVANCED BIQANALYTICAL SERVICES, INC. Figure 35. Full-Scan Mass Spectrum of Peak 6 in Figure 29 -Profile Q1 SCAN Fose Amide LC/MS-1/Scans 388-398 Scans 388-398 minus 1-200 & 420-712 Time=11.06 min Fose Amide LC/MS-1 - 7/5/95 - 4:32 PM 1 uL of Fose Amide(100 uM/top),delta C8,ACN/H20(5% to 95% in 15 min) 1 peak 95MYHW01.3M 1,269,023 43 005950 ADVANCED BIDANALYTICAL SERVICES, INC. Figure 36. Full-Scan Mass Spectrum of Peak 7 in Figure 29 Fose Amide LC/MS-1 /Scans 425-426 -Profile Q1SCAN Scans 425-426 minus 1-420 & 430-712 Time=11.98 min Fose Amide LC/MS-1- 7/5/95 - 4:32 PM 1 uL of Fose Amide(100 uM/top),delta C8,ACN/H20(5% to 95% in 15 min) 3 peaks 95MYHW01.3M 25,000 Relative Intensity (%) 44 05951 ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 37. TIC for the Injections of 1 jxL of Top, Middle and Bottom Day Zero Solution of Perfluorooctanamide 1001 Top 17,500,000 75' 50- Relative Intensity (%) 1001 Middle 25,400,000 Relative Intensity (%) 17,300,000 Relative Intensity (%) 201 301 5.6 8.5 Scan/Time (min) 45 401 11.3 501 14.1 005952 ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 38. TIC for the Injections of 1 |iL of Top, Middle and Bottom Day Seven Solution of Perfluorooctanamide 1001 Top 15,100,000 Relative Intensity (%) Relative Intensity (%) Relative Intensity (%) Scan/Time (min) 46 005953 ADVANCED BIOANALYTICAL SERVICES, INC. 95MYHW01.3M Figure 39. Extracted Ion Chromatograms for the Injection of 1 }iL of Day Zero Top Solution of Perfluorooctanamide C7F15CONH2 LC/MS-2- 7/5/95 - 6:25 PM 1 uL of C7F15CONH2(1 mM/top),delta C8,ACN/H20(5% to 95% in 15 min; 100/900 , TIC 990,000 369/369 297/297 l 220,000 20,000 311.5/311.5 tr\ i* CcO CD 325.5/325.5 5 I0T) 339.5/339.5 1 A 70,000 Jl 90,000 40,000 Jt 392/392 120,000 6 I K______________________________________________ _____________________ 412/412 l 0.0 1 2.5 90 370,000 7 ,A 5.0 7.5 10.0 12.5 15.0 178 267 354 443 531 Time (min)/Scan 47 0S 954 ADVANCED BIOANALYTICAL SERVICES, INC. Figure 40. Full-Scan Mass Spectrum of Peak 1 in Figure 39 C7F15CONH2 LC/MS-2/Scans 224-226 -Profile Q1SCAN Scans 224-226 minus 1-224 & 227-712 Time=6.32 min C7F15CONH2 LC/MS-2 - 7/5/95 - 6:25 PM 1 uL of C7F15CONH2(1 mM/top),delta C8,ACN/H20(5% to 95% in 15 min) 2 peaks 95MYHW01.3M 84,243 48 00595S ADVANCED BIOANALYTICAL SERVICES, INC. Figure 41. Full-Scan Mass Spectrum of Peak 2 in Figure 39 C7F15CONH2 LC/MS-2/Scans 164-167 -Profile Q1SCAN Scans 164-167 minus 1-140 & 200-712 Time=4.64 min C7F15CONH2 LC/MS-2 - 7/5/95 - 6:25 PM 1 uL of C7F15CONH2(1 mM/top),delta C8,ACN/H20(5% to 95% in 15 min) 4 peaks 95MYHW01.3M 221,740 49 005956 ADVANCED BIOANALYTICAL SERVICES. INC. Figure 42. Full-Scan Mass Spectrum of Peak 3 in Figure 39 C7F15CONH2 LC/MS-2/Scans 239-245 -Profile Q1 SCAN Scans 239-245 minus 1-235 & 250-500 Time=6.80 min C7F15CONH2 LC/MS-2 - 7/5/95 - 6:25 PM 1 uL of C7F15CONH2(1 mM/top),delta C8,ACN/H20(5% to 95% in 15 min) 2 peaks 95MYHW0I.3M 180,925 50 005957 ADVANCED BIOANALYTICASERVICES, INC. Figure 43. Full-Scan Mass Spectrum of Peak 4 in Figure 39 C7F15CONH2 LC/MS-2/Scans 251-261 -Profile Q1SCAN Scans 251-261 minus 1-240 & 360-712 Time=7.19 min C7F15CONH2 LC/MS-2 - 7/5/95 - 6:25 PM 1 uL of C7F15CONH2(1 mM/top),delta C8,ACN/H20{5% to 95% in 15 min) 2 peaks 95MYHW01.3M 169,107 51 005958 ADVANCED BIOANALYTICAL SERVICES, INC. Figure 44. Full-Scan Mass Spectrum of Peak 5 in Figure 39 C7F15CONH2 LC/MS-2/Scans 265-271 -Profile Q1 SCAN Scans 265-271 minus 1-262 & 274-712 Time=7.53 min C7F15CONH2 LC/MS-2 - 7/5/95 - 6:25 PM 1 uL of C7F15CONH2(1 mM/top),delta C8,ACN/H20(5% to 95% in 15 min) 2 peaks 95MYHW01.3M 45,726 52 005959 ADVANCED BIOANALYTICAL SERVICES, INC. Figure 45. Full-Scan Mass Spectrum of Peak 6 in Figure 39 C7F15CONH2 LC/MS-2/Scans 356-357 -Profile Q1 SCAN Scans 356-357 minus 1-354 & 370-712 Time=10.07 min C7F15CONH2 LC/MS-2 - 7/5/95 - 6:25 PM 1 uL of C7F15CONH2(1 mM/top),delta C8,ACN/H20(5% to 95% in 15 min) 3 peaks 95MYHW01.3M 105,000 53 005960 ADVANCED BIOANALYTICAL SERVICES, INC. Figure 46. Full-Scan Mass Spectrum of Peak 7 in Figure 39 C7F15CONH2 LC/MS-2/Scans 366-367 -Profile Q1 SCAN Scans 366-367 minus 1-362 & 380-712 Time=10.35 min C7F15CONH2 LC/MS-2- 7/5/95 - 6:25 PM 1 uL of C7F15CONH2(1 mM/top),delta C8,ACN/H20(5% to 95% in 15 min 2 peaks 95MYHW01.3M 329,613 Relative Intensity (%) 54 005961