液相色谱-三重四极杆串联质谱准确定量技术及实验优化研究—以一种新型利尿药为例

doi:10.5246/jcps.2022.03.016

中国药学(英文版) ›› 2022, Vol. 31 ›› Issue (3): 184-191.DOI: 10.5246/jcps.2022.03.016

液相色谱-三重四极杆串联质谱准确定量技术及实验优化研究—以一种新型利尿药为例

马雯¹^,^*(), 许悦², 王淑园², 李军¹, 杨宝学²^,^*()

1. 北京大学药学院天然药物及仿生药物国家重点实验室, 北京 100191
2. 北京大学基础医学院药理学系, 北京 100191

收稿日期:2021-10-23 修回日期:2021-11-24 接受日期:2021-12-30 出版日期:2022-03-31 发布日期:2022-03-31
通讯作者: 马雯, 杨宝学
作者简介:
+ Tel.: +86-10-82805103, E-mail: wen.ma@bjmu.edu.cn
+ E-mail: baoxue@bjmu.edu.cn
基金资助:
National Natural Science Foundation of China (Grant No. 22004004 and 81620108029).

Research on accurate quantitation technique using liquid chromatography coupled with triple quadrupole mass spectrometry and experiment optimization—take a novel diuretic as an example

Wen Ma¹^,^*(), Yue Xu², Shuyuan Wang², Jun Li¹, Baoxue Yang²^,^*()

1 State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
2 Department of Pharmacology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China

Received:2021-10-23 Revised:2021-11-24 Accepted:2021-12-30 Online:2022-03-31 Published:2022-03-31
Contact: Wen Ma, Baoxue Yang

摘要/Abstract

摘要：

液相色谱-三重四极杆串联质谱(LC-QQQ MS)技术是最常用的液质定量分析技术, 广泛应用于药理学、靶向代谢组学、中药质量控制等研究领域。该技术利用多反应监测(MRM)模式, 只对特定的母离子和子离子通道进行扫描监测, 能够在复杂基体中高特异性、高灵敏度检测到靶标分子。本研究以一种二芳基酰胺类利尿药为实验对象, 介绍液相色谱和质谱部分的方法建立和参数优化。利尿药和内标可以在5分钟梯度内完全分离, 定量线性范围为1–1000 ng/mL (R² = 0.9996)。优化结果表明, 液相色谱的流动相选择、洗脱梯度设置和质谱中化合物的去簇电压、碰撞能的设定是液相色谱-三重四极杆串联质谱方法学建立的关键。

关键词: 液相色谱, 三重四极杆串联质谱, 实验方法建立, 利尿药

Abstract:

Liquid chromatography coupled with the triple quadrupole mass spectrometry (LC-QQQ MS) technique is the most commonly used technique for quantitative analysis. It is widely used in pharmacology, targeted metabolomics, Chinese medicine quality control, and other research fields. The technique monitors only characteristic precursors and product ions through multiple reaction monitoring (MRM) mode and can detect targeted molecules in complex matrix with high specificity and sensitivity. In the present study, a diarylamide derivative diuretic was used as an example to introduce the method establishment and parameter optimization of this liquid chromatography-mass spectrometry technique. Diuretic and its internal standard could be completely separated within a 5-min gradient, and the quantitative linear range was 1–1000 ng/mL with R²= 0.9996 in practical samples. This study showed that the key to the method development for LC-QQQ MS was the selection of the LC mobile phase, the elution gradient, the declustering potential, and the collision energy of compounds in MS.

Key words: Liquid chromatography, Triple quadrupole mass spectrometry, Method establishment, Diuretic

Supporting:

马雯, 许悦, 王淑园, 李军, 杨宝学. 液相色谱-三重四极杆串联质谱准确定量技术及实验优化研究—以一种新型利尿药为例[J]. 中国药学(英文版), 2022, 31(3): 184-191.

Wen Ma, Yue Xu, Shuyuan Wang, Jun Li, Baoxue Yang. Research on accurate quantitation technique using liquid chromatography coupled with triple quadrupole mass spectrometry and experiment optimization—take a novel diuretic as an example[J]. Journal of Chinese Pharmaceutical Sciences, 2022, 31(3): 184-191.

图/表 7

Figure 1. (A) MRM mode in quantitation experiment; (B) structures of APAC and IS; (C) establishment procedure of quantitation method based on LC-MS/MS technique.

Figure 2. Optimization of DP, CE for APAC and IS.

Table 1. MS ion source parameter optimization.

Figure 3. Extracted ion chromatogram of APAC (1 ng/mL) when using (A) ACN containing 10% isopropanol and (B) pure ACN as mobile phase.

Figure 4. Influence of (A) different LC initial gradients and (B) equilibrium times on the results.

Table 2. RPLC-MS gradient for separation.

Figure 5. Plots of the relative mass intensity of APAC/IS vs. APAC concentrations.

参考文献 12

[1]	Lacorte, S.; Fernandez-Alba, A.R. Time of flight mass spectrometry applied to the liquid chromatographic analysis of pesticides in water and food. Mass Spectrom. Rev. 2006, 25, 866–880.
[2]	Li, X.J.; Li, H.M.; Ma, W.; Guo, Z.; Li, X.M.; Li, X.Q.; Zhang, Q.H. Determination of patulin in apple juice by single-drop liquid-liquid-liquid microextraction coupled with liquid chromatography-mass spectrometry. Food Chem. 2018, 257, 1–6.
[3]	Zhang, S.; Zhao, Y.; Wang, S.Y.; Li, M.; Xu, Y.; Ran, J.H.; Geng, X.Q.; He, J.Z.; Meng, J.; Shao, G.Y.; Zhou, H.; Ge, Z.M.; Chen, G.P.; Li, R.T.; Yang, B.X. Discovery of novel diarylamides as orally active diuretics targeting urea transporters. Acta Pharm. Sin. B. 2021, 11, 181–202.
[4]	Chen, S.L.; Kong, H.W.; Lu, X.; Li, Y.; Yin, P.Y.; Zeng, Z.D.; Xu, G.W. Pseudotargeted metabolomics method and its application in serum biomarker discovery for hepatocellular carcinoma based on ultra high-performance liquid chromatography/triple quadrupole mass spectrometry. Anal. Chem. 2013, 85, 8326–8333.
[5]	Yao, Q.Y.; Guo, Y.C.; Xue, J.S.; Kong, D.M.; Li, J.; Tian, X.Y.; Hao, C.Y.; Zhou, T.Y. Development and validation of a LC-MS/MS method for simultaneous determination of six glucocorticoids and its application to a pharmacokinetic study in nude mice. J. Pharm. Biomed. Anal. 2020, 179, 112980.
[6]	Cajka, T.; Fiehn, O. Toward merging untargeted and targeted methods in mass spectrometry-based metabolomics and lipidomics. Anal. Chem. 2016, 88, 524–545.
[7]	Qiao, X.; Ye, M.; Xiang, C.; Wang, Q.; Liu, C.F.; Miao, W.J.; Guo, D.A. Analytical strategy to reveal the in vivo process of multi-component herbal medicine: a pharmacokinetic study of licorice using liquid chromatography coupled with triple quadrupole mass spectrometry. J. Chromatogr. A. 2012, 1258, 84–93.
[8]	Ma, W.; Li, J.; Li, X.J.; Liu, H.W. Enrichment of diamide insecticides from environmental water samples using metal-organic frameworks as adsorbents for determination by liquid chromatography tandem mass spectrometry. J. Hazard. Mater. 2022, 422, 126839.
[9]	Knepper, M.A.; Miranda, C.A. Urea channel inhibitors: a new functional class of aquaretics. Kidney Int. 2013, 83, 991–993.
[10]	Wang, S.Y.; Xu, Y.; Zhao, Y.; Zhang, S.; Li, M.; Li, X.W.; He, J.Z.; Zhou, H.; Ge, Z.M.; Li, R.T.; Yang, B.X. N-(4-acetamidophenyl)-5-acetylfuran-2-carboxamide as a novel orally available diuretic that targets urea transporters with improved PD and PK properties. Eur. J. Med. Chem. 2021, 226, 113859.
[11]	Zhang, Z.Y.; Wang, X.; Liu, D.; Zhang, H.; Zhang, Q.; Lu, Y.Y.; Li, P.; Lou, Y.Q.; Yang, B.X.; Lu, C.; Lou, Y.X.; Zhang, G.L. Development and validation of an LC-MS/MS method for the determination of a novel thienoquinolin urea transporter inhibitor PU-48 in rat plasma and its application to a pharmacokinetic study. Biomed. Chromatogr. 2018, 32, e4157.
[12]	Zhang, Z.P.; Tang, W. Drug metabolism in drug discovery and development. Acta Pharm. Sin. B. 2018, 8, 721–732.

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液相色谱-三重四极杆串联质谱准确定量技术及实验优化研究—以一种新型利尿药为例

Research on accurate quantitation technique using liquid chromatography coupled with triple quadrupole mass spectrometry and experiment optimization—take a novel diuretic as an example

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