实验6 兔血清中茶碱的高效液相色谱法测定ppt参考课件.ppt

Experiment Ⅵ Determination of theophylline in rabbit plasma by HPLC * * 1. Purpose (1) To learn the serum sample treatment method and to be familiar with the blood sample collection method (2) To learn the experiment on the determination of theophylline in rabbit serun by HPLC * * 2. Introduction Theophylline Caffine N O N H3C—N N O CH3 H * * 3. Procedure and Method (1) Preparation of reference standards solution Theophylline （THP）standard solution: 1.0 mg/mL (2) Preparation of internal standard solution Caffeine (CAF): 1.0 ?g/mL (Prepared already) (Prepared already) * * (3) Standard solution (Prepared already) 50.0, 125.0, 250.0, 500.0 and 1000.0 ?g/mL (i) Theophylline standard solution: (ii) Theophylline serum sample prepareartion * * (3) Standard solution (Prepared already) 0, 50.0, 125.0, 250.0, 500.0 and 1000.0 ?g/mL (i) Theophylline standard solution: (ii) Theophylline serum sample preparation * * The calibration curve was constructed by plotting the peak area ratios of each analyte to the IS versus the concentration using weighed linear least square regression. (4) Calibration curve * * (5)HPLC conditions Column:ODS column (5?m, 150mm×4.6mm i.d) Mobile phase：VCH3OH/VH2O=35/65 Flow rate: 1.0 mL min-1 Wavelength: 254 nm Injection volumn: 20 ?L 温馨提示：高效液相色谱在本层楼的东侧403房间 结束后值日生记得清扫卫生哦！ * * Results Calibration curve , the linear equation and R NO. CTHP THP CAF ATHP/ACAF tR A tR A 空白 1 2 3 4 5 * * 4. Attention * * * * * *