色谱柱和色谱系统故障检修.ppt

**olumnandSystemTroubleshooting色谱柱和色谱系统的故障检修WhatEveryHPLCUserShouldKnow每个HPLC使用者应该知道什么**ponentsHPLC的组成Pump泵Injector/Autosampler进样器/自动进样器Column色谱柱Detector检测器DataSystem/Integrator数据系统/。****PressureIssues压力问题ColumnObservationsPotentialProblems色谱柱观测潜在的问题HighpressurePluggedfrit压力高堵塞滤头Columncontamination色谱柱污染Piuggedpacking堵塞填料**DeterminingtheCauseandCorrectingHighBackPressure查出原因纠正压力Checkpressurewith/withoutcolumn-uard有/无柱子时检查压力-许多压力问题是由于系统或保护柱的阻塞IfColumnPressureishigh如果柱压高:Washcolumn-Eliminatecolumncontaminationand冲洗柱子pluggedpacking排除柱污染和填料堵塞-highmolecularweight/pounds高分子量/被吸附化合物-precipitatefromsampleorbuffer样品或缓冲液中的沉淀物Backflushcolumn-Clearpluggedfrit反冲柱子-清洁阻塞的滤头Changefrit-Clearpluggedfrit更换滤头**ColumnCleaning柱子清洗Flushwithstrongersolventsthanyourmobilephase用比你的流动相更强的溶剂冲洗Reversed-PhaseSolventChoicesinOrderofIncreasingStrength为了增加强度选择反相溶剂Useatleast25mlofeachsolventforanalyticalcolumns对于分析柱每种溶剂至少用25ml冲洗Mobilephasewithoutbuffersalts没有缓冲盐的流动相100%Methanol甲醇100%Acetonitrile乙***75%Acetonitrile:25%Isopropanol75乙***:25%异丙醇100%Isopropanol异丙醇100%MethyleneChloride★二***甲烷100%Hexane★已烷★WhenusingeitherHexaneorMethyleneChloridethecolumnmustbeflushedwithIsopropanolbeforereturningtoyourresvered-***甲烷柱子时,必须先用异丙醇冲洗,然后再用你的反相流动相**ColumnCleaning柱子清洗NormalPhaseSolventChoicesinOrderofIncreasingStrength为了增加强度选用正相溶剂Useatleast50mlofeachsolvent每种溶剂至少用50ml50%Methanol:50%Chloroform50%甲醇:50%***仿100%EthylAcetate乙酸乙酯**HowtoChangeaFritpressionFerrule压缩金属套圈Weargloves戴手套Donotallowbedtodry不要让底座干燥Donottouchthecolumn-bodyheatwillextrudepacking不要使柱身受热,否则填料会喷出Donotovertighten不要旋得太紧FemaleEndFitting旋好尾端螺母MaleEndFitting旋好尾端螺头**PreventingBackPressureProblems压力问题的预防Usecolumnprotection柱保护-Guardcolumns保护柱/预柱-In-linefilters在线过滤器SamplePreparation样品预处理Appropriatecolumnflushing适当的柱冲洗Filterbufferedmobilephase过滤缓冲流动相**PreventingBackPressureProblems:In-LineDevices压力问题的预防:在