1 HPLC Column and  System Troubleshooting 色谱柱和色谱系统的故障检修   What Every HPLC User Should Know   每个HPLC使用者应该知道什么  HPLC Components HPLC 的组成     Pump 泵     Injector/Autosampler 进样器/自动进样器     Column 色谱柱     Detector 检测器     Data System/Integrator 数据系统/积分仪表   All of these components can have problems and require troubleshooting.  所有这些部件可能出现故障而需要检修。 Categories of Column Problems  色谱柱问题的类型        A. Pressure 压力    B. Peak shape 峰形    C. Retention 保留时间 Pressure Issues压力问题  Column Observations       Potential Problems 色谱柱观测                            潜在的问题        High pressure                    Plugged frit   压力高                                  堵塞滤头                                            Column contamination                                            色谱柱污染                                            Piugged packing                                            堵塞填料 Determining the Cause and Correcting High Back Pressure   查出原因 纠正压力 Check pressure with/without column - many pressure problems are due to blockages in the system or guard 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 If Column Pressure is high 如果柱压高: Wash column - Eliminate column contamination and  冲洗柱子           plugged packing  排除柱污染和填料堵塞                      - high molecular weight/adsorbed                        compounds  高分子量／被吸附化合物                      - precipitate from sample or buffer                         样品或缓冲液中的沉淀物 Back flush column - Clear plugged frit    反冲柱子－清洁阻塞的滤头 Change frit - Clear plugged frit    更换滤头 Column Cleaning 柱子清洗 F lush with stronger solvents than your mobile phase 用比你的流动相更强的溶剂冲洗    Reversed-Phase Solvent Choices in Order of     Increasing Strength    为了增加强度选择反相溶剂 Use at least 25 ml of each solvent for analytical columns                                   对于分析柱每种溶剂至少用25 ml 冲洗 Mobile phase without buffer salts  没有缓冲盐的流动相 100% Methanol  甲醇 100% Acetonitrile  乙腈 75% Acetonitrile :25% Isopropanol  75乙腈 ： 25％异丙醇 100% Isopropanol  异丙醇 100% Methylene Chloride★  二氯甲烷 100% Hexane ★  已烷 ★  When using either Hexane or Methylene Chloride the column must be flushed with  Isopropanol before returning to your resvered-phase mobile phase. 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 Column Cleaning柱子清洗 Normal Phase Solvent Choices in Order of Increasing Strength 为了增加强度选用正相溶剂  Use at least 50 ml of each solvent            每种溶剂至少用50 ml   50% Methanol : 50% Chloroform              50% 甲醇 : 50% 氯仿 100% Ethyl Acetate    乙酸乙酯 How to Change a Frit 怎样更换滤头 Column Inlet Frit 柱进口滤头 Column Body 柱身 Compression Ferrule 压缩金属套圈 Wear gloves 戴手套 Do not allow bed to dry 不要让底座干燥 Do not touch the column-body heat will extrude packing       不要使柱身受热,否则填料会喷出 Do not over tighten 不要旋得太紧 Female End Fitting 旋好尾端螺母 Male End Fitting 旋好尾端螺头 Preventing Back Pressure Problems压力问题的预防 Use column protection 柱保护  - Guard columns 保护柱/预柱  - In-line filters 在线过滤器  Sample Preparation 样品预处理 Appropriate column flushing                       适当的柱冲洗 Filter buffered mobile phase                      过滤缓冲流动相 Preventing Back Pressure Problems:In-Line Devices 压力问题的预防:在线装置 Mobile Phase    Pre-Column    Injector  F
色谱柱和色谱系统的故障检修.ppt