文档介绍：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
请访问中国色谱网()
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1
ponents 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 ponents can have problems and require troubleshooting.
所有这些部件可能出现故障而需要检修。
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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
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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
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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
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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***
75% Acetonitrile :25% Isopropanol 75乙***: 25%异丙醇
100% Isopropanol 异丙醇
100% Methylene Chloride★二***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***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
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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%