1. Presented by Meng Shi The C holesterol in plasma was a important factor to lead Thrombus

2. Presented by Meng Shi The C holesterol in plasma was a important factor to lead Thrombus The Cholesterol was tested as an independent predictor of Thrombus formation 1. Therefore, to study the cholesterol level and some other reverent component in plasma was meaningful in studding about the formation of thrombus 1. Tasneem Z. The American Journal of Cardiology Vol. 84 November 1, 1999

3. Presented by Meng Shi The C holesterol in plasma was a important factor to lead Thrombus Problem The problem was the relationship about cholesterol level in plasma and the thrombus. Moreover, the additional problem was the detection method of the plasma sample. Hypothesis: The Cholesterol in plasma was a important factor to lead Thrombus

4. Presented by Meng Shi The C holesterol in plasma was a important factor to lead Thrombus Research Strategy(proposed data analysis). Concentration of Cholesterol in plasma versus the statistic number of apoplexy patient. Concentration of other component, ions, in plasma versus the statistic number of apoplexy patient. If two were correlated. Mechanism study.

5. Presented by Meng Shi The C holesterol in plasma was a important factor to lead Thrombus Method of Isolating Cholesterol in Plasma # to 0.8 volume parts plasma sample 3 parts of a chloroform/methanol (1:2 mixture (v/v)) are added to give one phase # after vigorously mixing the sample is kept for about an hour at room temperature # add 1 volume part each chloroform and water for phases separation # after mixing the sample is centrifuged at high speed for 15 min # the lower chloroform phase containing the Cholesterol is isolated

6. Presented by Meng Shi The C holesterol in plasma was a important factor to lead Thrombus Isolation There were two candidate isolation method, HPLC and GC. HPLC. Higher resolution, but slower, solvent effect. GC. Faster, relative lower resolution, gas phase. Final decision Since HPLC could provide a liquid phase that helped to using UV-visible spectrum. The HPLC was chosen

7. Presented by Meng Shi The C holesterol in plasma was a important factor to lead Thrombus Detection Method Comparison. IR: 3100 cm for aromatic compounds Not quantitated Fluorescence: The emission wavelength was 470-490nm 3, and the excitation wavelength was 325 and 308 nm. The LOD was 10nM 4 Mass Spectrum; MS-MS with a ESI ionizer The LOD was 0.45 uM 2 suit for the expected analysis. 2. Joni J., Journal of Chromatography B, 821 (2005) 144–152. 3. I. Maarek et.al Photochemistry and Photobiology, 2000, 71(2): 178–187 4. Fernando V., Anal. Chem. 2010, 82, 6775–6781

8. Presented by Meng Shi The C holesterol in plasma was a important factor to lead Thrombus Instrument HPLC was chosen, since the cholesterol should be quantitive measured. HPLC: Agilent. 920-LC gradient analytical system. (Part No. 001009050X) column: Solvent Miser C18, 5µm, 250 x 2.1mm column (Part No. 9945) Detector: Agilent 385-LC cooled evaporative light scattering detector (external),. (Part No. PL08901110 ) 5

9. Presented by Meng Shi The C holesterol in plasma was a important factor to lead Thrombus Other component. There were other component that should be studied in plasma. Most of them was metal ion such as sodium ion, potassium ion, and calcium ion. The good method would be ICE-APS, which was suit to deal with biology sample. The LOD would be 0.05mg/L 4 4. Michaela Z, Microchemical Journal 81 (2005) 171– 176.

10. Presented by Meng Shi The C holesterol in plasma was a important factor to lead Thrombus Instrument and Sample Data 6. ROBERT T., Chemistry and Physics of Lipids, 36 (1984) 1-14. Agilent 920-LC Analytical HPLC system Sample Fluorescence spectrum of Cholesterol, where the A part represented the absorbance, and B represented the corrected fluorescence. 6

11. Presented by Meng Shi The C holesterol in plasma was a important factor to lead Thrombus Further problem Could ion be analysis through HPLC? The metal ion has nearly no retention time and there no difference in stationary phase in HPLC New stationary phase? Metallopolymers 7 7. George R. Adv. Mater. 2007, 19, 3439–3468

12. Presented by Meng Shi The C holesterol in plasma was a important factor to lead Thrombus Conclusion The cholesterol was a independent predictor towards thrombus formation. If other component in plasma contributing to the formation of Thrombus The use of Metallopolymers as stationary phase to help the detection of metal ion and complex