高效液相色谱法是天然化合物定性定量的重要方法之一,高效液相色谱仪也是一类贵重仪器。介绍了高效液相色谱仪的分离原理、特点及其定性定量的方法,样品前处理方法;同时在实验教学中,建立HPLC内标法测定茶叶中咖啡因的含量。通过本实验学生不仅能掌握样品前处理方法、还能更好地理解和掌握HPLC的分离原理、操作方法、定性、定量方法及其在天然化合物分析研究中的重要性;通过实验教学中每人一份样品,提高了学生的学习兴趣,增强了其动手能力;此外该实验还特别培养了学生对贵重仪器的维护与保养意识。
High performance liquid chromatography(HPLC) method is one of the important methods for native compound analysis.This paper introduces the separation principle,characteristics and the qualitative and quantitative methods of HPLC and sample preparation methods.In the experimental teaching,the determination method of the content of caffeine in tea was established utilizing HPLC by taking benzyl alcohol as an internal standard.Through this experiment,students not only learn the method of sample preparation,but also better understand and master the analysis principles,operation methods,qualitative and quantitative methods,and importance of HPLC in the analysis of natural compounds.One sample is tested by one student in the experimental teaching,which increases students' learning interest and enhances their ability.In addition,this experiment also cultivated students' awareness in maintaining valuable instruments.