色谱柱和色谱系统故障.ppt

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1、HPLCColumnandSystemTroubleshooting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2021/8/51HPLCComponentsHPLC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/积分仪表Allofthesecomponentscanhaveproblemsandrequiretroubleshooting.所有这些部件可能

2、出现故障而需要检修。2021/8/52CategoriesofColumnProblems色谱柱问题的类型A.Pressure压力B.Peakshape峰形C.Retention保留时间2021/8/53PressureIssues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2021/8/54DeterminingtheCauseandCorre

3、ctingHighBackPressure查出原因纠正压力Checkpressurewith/withoutcolumn-manypressureproblemsareduetoblockagesinthesystemorguard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/adsorbedc

4、ompounds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2021/8/55ColumnCleaning柱子清洗Flushwithstrongersolventsthanyourmobilephase用比你的流动相更强的溶剂冲洗Reversed-PhaseSolventChoicesinOrderofIncreasingStreng

5、th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乙腈75%Acetonitrile:25%Isopropanol75乙腈：25％异丙醇100%Isopropanol异丙醇100%MethyleneChloride★二氯甲烷100%Hexane★已烷★WhenusingeitherHexaneorMe

6、thyleneChloridethecolumnmustbeflushedwithIsopropanolbeforereturningtoyourresvered-phasemobilephase.当用已烷蔌二氯甲烷柱子时,必须先用异丙醇冲洗,然后再用你的反相流动相2021/8/56ColumnCleaning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

7、:50%Chloroform50%甲醇:50%氯仿100%EthylAcetate乙酸乙酯2021/8/57HowtoChangeaFrit怎样更换滤头ColumnInletFrit柱进口滤头ColumnBody柱身CompressionFerrule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

8、螺母MaleEndFitting旋好尾端螺头2021/8/58PreventingBackPressureProblems压力问题的预防Usecolumnprotection柱保