Simultaneous determination of nucleosides, modified nucleosides and mononucleotides in gastric cancer cells by liquid chromatography coupled with tandem mass spectrometry

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-09-30 DOI:10.1016/j.jpba.2025.117177

Juan Li , Piao Zhou , Manni He , Mengxue Liu , Duo Chen , Hongmin Liu

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Abstract

Nucleosides and their modified forms serve as fundamental building blocks of nucleic acids and play critical roles in epigenetic regulation, RNA metabolism, and cellular signaling. In this study, we developed and validated a rapid, sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous quantification of 27 endogenous nucleosides—including 12 canonical nucleosides, 11 modified nucleosides, and 4 mononucleotides—in cellular samples. Under optimized chromatographic conditions, good separations and peak shapes for 27 target compounds were achieved within a 13.0-min gradient elution. The overall LOQs were between 0.2 and 25.0 ng/mL. The intra-day precision of the method was less than 8.2 %, and the inter-day precision was less than 14.2 %. The accuracy was in the range of 89.9–112.2 % for all analytes. The developed method was successfully applied to analyze the metabolic profiles of gastric cancer cells with varying differentiation degrees (NCI-N87, SGC-7901, MGC-803). The results revealed distinct nucleotide metabolism alterations linked to tumor progression and identified potential biomarkers for gastric cancer advancement.

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液相色谱-串联质谱法同时测定胃癌细胞中的核苷、修饰核苷和单核苷酸。

核苷及其修饰形式是核酸的基本组成部分，在表观遗传调控、RNA代谢和细胞信号传导中发挥着关键作用。在这项研究中，我们建立并验证了一种快速、灵敏的超高效液相色谱-串联质谱（UHPLC-MS/MS）方法，用于同时定量细胞样品中的27种内源性核苷，包括12种典型核苷，11种修饰核苷和4种单核苷酸。在优化的色谱条件下，27个目标化合物在13.0 min的梯度洗脱时间内获得了良好的分离效果和峰形。总体loq在0.2 ~ 25.0 ng/mL之间。该方法日内精密度小于8.2% %，日内精密度小于14.2% %。所有分析物的准确度为89.9-112.2 %。该方法已成功应用于不同分化程度胃癌细胞（NCI-N87、SGC-7901、MGC-803）的代谢谱分析。结果显示，不同的核苷酸代谢改变与肿瘤进展有关，并确定了胃癌进展的潜在生物标志物。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of pharmaceutical and biomedical analysis 医学-分析化学

CiteScore

6.70

自引率

5.90%

发文量

588

审稿时长

37 days

期刊介绍： This journal is an international medium directed towards the needs of academic, clinical, government and industrial analysis by publishing original research reports and critical reviews on pharmaceutical and biomedical analysis. It covers the interdisciplinary aspects of analysis in the pharmaceutical, biomedical and clinical sciences, including developments in analytical methodology, instrumentation, computation and interpretation. Submissions on novel applications focusing on drug purity and stability studies, pharmacokinetics, therapeutic monitoring, metabolic profiling; drug-related aspects of analytical biochemistry and forensic toxicology; quality assurance in the pharmaceutical industry are also welcome. Studies from areas of well established and poorly selective methods, such as UV-VIS spectrophotometry (including derivative and multi-wavelength measurements), basic electroanalytical (potentiometric, polarographic and voltammetric) methods, fluorimetry, flow-injection analysis, etc. are accepted for publication in exceptional cases only, if a unique and substantial advantage over presently known systems is demonstrated. The same applies to the assay of simple drug formulations by any kind of methods and the determination of drugs in biological samples based merely on spiked samples. Drug purity/stability studies should contain information on the structure elucidation of the impurities/degradants.