色谱柱和色谱系统的故障检修.pptx

HPLC Column and System Troubleshooting 色谱柱和色谱系统的故障检修What Every HPLCUser 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 Inl