超高效液相色谱与高效液相色谱方法转换及验证

Method conversion and validation between HPLC and UPLC

超高效液相色谱(UPLC)分离原理与高效液相色谱(HPLC)完全一致,但分离度更好,灵敏度更高,并且速度快,消耗溶剂少。如果实现两者方法的转换,并进行必要的验证工作,可在不变更标准的前提下合规应用UPLC技术,对药品质量进行高效精准的控制。中国药典2015年版四部关于UPLC的应用有简单说明,但是涉及与HPLC方法的转换和验证没有细节规定,使该技术推广应用受阻。本文从2种仪器的差异进行分析,阐述了方法转换时如何进行参数变更,包括色谱柱、流速、进样体积和梯度洗脱程序的转换。基于转换前方法已经经过全面验证,转换后的方法验证应主要着眼于仪器带来的差异,需验证分离度和灵敏度2个方面。本文根据转换前方法的不同情况,对方法验证细节要求进行了分析讨论,以确保方法转换后准确有效,合乎规范。

Ultra-high performance liquid chromatography（UPLC）and high performance liquid chromatography（HPLC）share the same basic principle.However,compared to HPLC,UPLC provides separation with higher resolution at higher speed with reduced solvent consumption.The increased demand on high efficiency drug quality control has made the conversion between HPLC and UPLC an obvious task,yet guidance of validation has not been stated.The 2015 edition of the Chinese pharmacopeia（ChP）part Ⅳ offered a brief description about the application of UPLC;however,no specific requirements was elaborated for the conversion methodology and the validation/verification process between HPLC and UPLC.In this article,we compared the differences between HPLC and UPLC and explained how to modify the parameters during the conversion such as column packing,flow rate,inject volume and gradient process etc.Considering the fact that original method has been thoroughly validated,the necessary validation for method after conversion should focus on the separation accuracy and sensitivity which could be solely affected by the equipment differences.Based on the different situation of original method,this paper discussed the specific requirements of validation to ensure the accuracy,efficiency and compliance of the method after conversion.