基于液相色谱分离技术的核酸类药物生物分析方法研究进展

孙淑萌; 林琳; 张岱州; 李欣; 关永霞; 陈凯

doi:10.6039/j.issn.1001-0408.2024.23.20

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基于液相色谱分离技术的核酸类药物生物分析方法研究进展

综述 | 更新时间：2024-12-09

- 基于液相色谱分离技术的核酸类药物生物分析方法研究进展
- Liquid chromatography-based bioanalytical technologies for nucleic acid drugs
- 中国药房 2024年35卷第23期页码：2959-2964
- 作者机构：
  
  1.山东省药学科学院，济南　250101
  2.山东省人工噬菌体药物技术创新中心，济南　250101
  3.鲁南制药集团股份有限公司，山东临沂　276000
  4.山东中医药大学中医药创新研究院，济南　250001
- 作者简介：
  
  工程师，硕士。研究方向：新药药代动力学。电话：0531-88562151。E-mail：sunshumeng.123@163.com
  高级工程师，硕士生导师，博士研究生。研究方向：新药药代动力学。电话：0531-88562177。E-mail：chenkai@sdaps.cn
- 基金信息：
  
  山东省重点研发计划（重大科技创新工程）项目(2021CXGC010515)
- DOI：10.6039/j.issn.1001-0408.2024.23.20
  中图分类号： R917
- 纸质出版日期：2024-12-15，
  
  收稿日期：2024-04-17，
  
  修回日期：2024-11-19，
- 稿件说明：
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孙淑萌,林琳,张岱州等.基于液相色谱分离技术的核酸类药物生物分析方法研究进展 ^Δ[J].中国药房,2024,35(23):2959-2964.

SUN Shumeng,LIN Lin,ZHANG Daizhou,et al.Liquid chromatography-based bioanalytical technologies for nucleic acid drugs[J].ZHONGGUO YAOFANG,2024,35(23):2959-2964.
孙淑萌,林琳,张岱州等.基于液相色谱分离技术的核酸类药物生物分析方法研究进展 ^Δ[J].中国药房,2024,35(23):2959-2964. DOI： 10.6039/j.issn.1001-0408.2024.23.20.

SUN Shumeng,LIN Lin,ZHANG Daizhou,et al.Liquid chromatography-based bioanalytical technologies for nucleic acid drugs[J].ZHONGGUO YAOFANG,2024,35(23):2959-2964. DOI： 10.6039/j.issn.1001-0408.2024.23.20.

摘要

核酸类药物的生物分析方法主要有基于配体结合分析、基于聚合酶链反应和基于液相色谱分离技术3大类，前两者灵敏度虽高，但选择性较差，很难区分寡核苷酸本身及其较短的代谢产物。基于液相色谱分离技术生物分析方法的灵敏度虽略有逊色，但由于其具有高选择性，能够区分寡核苷酸本身及其代谢产物，故在核酸类药物的临床前和临床研究中展现出了广阔的应用前景。本文综述了该类方法中的高效液相色谱-紫外检测（HPLC-UV）法、高效液相色谱-荧光（HPLC-FL）法、液相色谱-串联质谱（LC-MS/MS）法、液相色谱-高分辨质谱法、微流液相色谱-串联质谱（microflow LC-MS/MS）法、杂交液相色谱-串联质谱法的特点及其在核酸类药物分析中的应用情况，发现除HPLC-UV法的检测灵敏度较低以外，其余方法均具有较高的灵敏度，但尚存在一些不足，如HPLC-FL法需要设计合适的探针、LC-MS/MS法需要标准物质、microflow LC-MS/MS法成本较高等。另外，一些相关策略及技术（如非特异性吸附解决策略、样品前处理技术等）的发展，在提高方法灵敏度的同时，也进一步加速了基于液相色谱分离技术的核酸类药物生物分析方法的发展。

Abstract

There are three types of bioanalytical methods for nucleic acid drugs， including ligand binding assay， quantitative polymerase chain reaction and liquid chromatography-based bioanalytical technologies. Although the first two assays have high sensitivity， they have poor selectivity and can not differentiate between intact and truncated metabolites. Liquid chromatography-based bioanalytical technologies which are less sensitive， offer high selectivity for the identification of intact and truncated metabolites. They have broad application prospects in both preclinical and clinical investigations of therapeutic nucleic acid drugs. This paper provides a critical review on the characteristics of these technologies and their application to analyze nucleic acid drugs， including high performance liquid chromatography-ultraviolet detection （HPLC-UV）， high performance liquid chromatography-fluorescence （HPLC-FL）， liquid chromatography-tandem mass spectrometry （LC-MS/MS）， liquid chromatography-high resolution-mass spectrometry， microflow liquid chromatography-tandem mass spectrometry （microflow LC-MS/MS） and hybridization liquid chromatography-tandem mass spectrometry. Although these technologies have high sensitivity except for HPLC-UV， they still have some shortcomings， such as suitable probes need to be designed for HPLC-FL， standard substance for LC-MS/MS， and high cost for microflow LC-MS/MS. In addition， the development of some related strategies or technologies （e.g. non-specific adsorption strategy， sample pretreatment） which can improve the sensitivity， has hastened the development of liquid chromatography-based bioanalytical technologies for nucleic acid drugs.

关键词

核酸类药物寡核苷酸生物分析液相色谱分离技术

Keywords

oligonucleotidebioanalysisliquid chromatographyseparation technology

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