色谱柱和色谱系统故障排除.pptVIP

* * 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% Hexan