【摘要】 基于液相色谱-高分辨质谱联用技术（LC-HRMS）整合代谢组学与脂质组学分析的方法已成为生物标志物筛选的重要技术手段。细胞外囊泡（EVs）是广泛存在于生物体液中的脂质双层包被的纳米级颗粒，凭借其内容物稳定性及与病理生理状态的高度关联性，已成为癌症生物标志物的重要载体。本研究构建了一种基于双色谱柱串联的极性扩展液相色谱系统（Polarity extended liquid chromatography, PELC）。此系统经一次进样过程即可实现对宽极性范围（涵盖极性与非极性）化合物的高效覆盖，同时有效降低了分析变异性。本研究首次将PELC-HRMS技术应用于EVs的代谢组-脂质组同步分析。结合MS-DIAL软件及人类代谢组数据库（Human Metabolome Database）进行代谢物注释与鉴定，从尿液EVs中共鉴定出193种代谢物。通过多元统计分析筛选出17种在前列腺癌中丰度存在显著差异的代谢物。这些差异代谢物主要富集于嘌呤代谢、淀粉与蔗糖代谢、半乳糖代谢、半胱氨酸与甲硫氨酸代谢以及不饱和脂肪酸生物合成等通路，提示这些代谢通路在前列腺癌中可能发生紊乱。其中，对肌酸（AUC=0.92）与甘油二酯（AUC=0.80）均表现出优异的诊断效能。本研究结果表明，PELC技术可作为复杂生物样本多组学整合分析的可靠平台，为深入解析代谢重编程机制及相关疾病的病理生理学提供了有效工具。更多还原

【Abstract】 Integrated metabolomic and lipidomic profiling, utilizing liquid chromatography coupled with highresolution mass spectrometry(LC-HRMS),has emerged as a pivotal strategy for biomarker discovery. However, the inherent polarity disparity between metabolites and lipids complicates simultaneous analysis. To address this, a dual-stationary phase polarity-extended liquid chromatography(PELC) system was developed, which surpassed conventional one-dimensional LC(1D-LC) by enabling comprehensive coverage of both polar and non-polar compounds within a single injection. This system enhanced chromatographic resolution, peak capacity, and throughput while minimizing analytical variability. Extracellular vesicles(EVs), lipid bilayer-enclosed nanoparticles ubiquitously present in biofluids, had gained prominence as reservoirs of cancer biomarkers due to their cargo stability and pathophysiological relevance. Herein, the application of PELC-HRMS for concurrent metabolome-lipidome profiling in EVs was pioneered. A total of 193 metabolites were identified using this technique coupled with MS-DIAL software and Human Metabolome Database. Subsequently, this technique was employed to explore potential biomarkers for prostate cancer(PCa). Multivariate analysis identified 17 differentially abundant metabolites in PCa, implicating dysregulated pathways including purine metabolism, starch and sucrose metabolism, galactose metabolism, cysteine and methionine metabolism, and biosynthesis of unsaturated fatty acids. Notably, creatine(AUC=0.92) and DG 42:5(AUC=0.80) demonstrated robust diagnostic efficacy, attributable to their broad polarity ranges and EV-specific enrichment. This study established PELC as a high-fidelity platform for multi-omics integration in complex biospecimens, advancing mechanistic insights into metabolic rewiring and disease pathophysiology.更多还原