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1、Pharmaceutical Analysis2012-11Experiment Determination of theophylline in rabbit plasma by HPLCPharmaceutical Analysis1. 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 i
2、n rabbit serun by HPLCPharmaceutical Analysis2. Introduction Theophylline CaffineNONH3CNNOCH3HPharmaceutical Analysis3. Procedure and Method (1) Preparation of reference standards solutionTheophylline （THP）standard solution: 1.0 mg/mL(2) Preparation of internal standard solution Caffeine (CAF): 1.0
3、g/mL (Prepared already)(Prepared already)Pharmaceutical Analysis(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 prepareartionPharmaceutical Analysis(3) Standard solution(Prepared already)0, 50.0, 125.0
4、, 250.0, 500.0 and 1000.0 g/mL(i) Theophylline standard solution: (ii) Theophylline serum sample preparationPharmaceutical AnalysisThe 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
5、) Calibration curve Pharmaceutical Analysis(5)HPLC conditions Column:ODS column (5 m, 150mm4.6mm i.d)Mobile phase：VCH3OH/VH2O=35/65 Flow rate: 1.0 mL min-1Wavelength: 254 nmInjection volumn: 20 L 温馨提示：高效液相色谱在本层楼的东侧温馨提示：高效液相色谱在本层楼的东侧403房间房间 结束后值日生记得清扫卫生哦！结束后值日生记得清扫卫生哦！Pharmaceutical AnalysisResultsCalibration curve , the linear equation and RNO.CTHPTHPCAFATHP/ACAFtRAtRA空白空白12345Pharmaceutical Analysis4. Attention 高效液相色谱高效液相色谱403房间房间高效液相色谱高效液相色谱403房间房间Pharmaceutical Analysis2012-11实验结束后实验结束后 请您照上图摆放好请您照上图摆放好
