实验6 兔血清中茶碱的高效液相色谱法测定.ppt

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1、2012-11ExperimentⅥDeterminationoftheophyllineinrabbitplasmabyHPLC1.Purpose(1)Tolearntheserumsampletreatmentmethodandtobefamiliarwiththebloodsamplecollectionmethod(2)TolearntheexperimentonthedeterminationoftheophyllineinrabbitserunbyHPLC2.IntroductionTheophyllineCaffineNO

2、NH3C—NNOCH3H3.ProcedureandMethod(1)PreparationofreferencestandardssolutionTheophylline（THP）standardsolution:1.0mg/mL(2)PreparationofinternalstandardsolutionCaffeine(CAF):1.0g/mL(Preparedalready)(Preparedalready)(3)Standardsolution(Preparedalready)50.0,125.0,250.0,500.0a

3、nd1000.0g/mL(i)Theophyllinestandardsolution:(ii)Theophyllineserumsampleprepareartion(3)Standardsolution(Preparedalready)0,50.0,125.0,250.0,500.0and1000.0g/mL(i)Theophyllinestandardsolution:(ii)TheophyllineserumsamplepreparationThecalibrationcurvewasconstructedbyplottin

4、gthepeakarearatiosofeachanalytetotheISversustheconcentrationusingweighedlinearleastsquareregression.(4)Calibrationcurve(5)HPLCconditionsColumn:ODScolumn(5m,150mm×4.6mmi.d)Mobilephase：VCH3OH/VH2O=35/65Flowrate:1.0mLmin-1Wavelength:254nmInjectionvolumn:20L温馨提示：高效液相色谱在本层楼

5、的东侧403房间结束后值日生记得清扫卫生哦！ResultsCalibrationcurve,thelinearequationandRNO.CTHPTHPCAFATHP/ACAFtRAtRA空白123454.Attention高效液相色谱403房间高效液相色谱403房间ThankYou!2012-11实验结束后请您照上图摆放好