基于脂质组学的冬虫夏草特异性脂质标志物筛选

目的：比较野生冬虫夏草（YS）、人工栽培冬虫夏草（RG）及3种常见混伪品（戴氏虫草DS、凉山虫草LS、亚香棒虫草YXB）的脂质组成差异，筛选并鉴定能够精准区分野生冬虫夏草的特异性脂质标志物，为冬虫夏草的真伪鉴别与质量评价提供科学依据。方法：采用非靶向脂质组学策略，使用改良MTBE法提取5类虫草样本中的总脂质，利用超高效液相色谱-电喷雾电离串联质谱（UHPLC-ESI-MS/MS）技术进行脂质图谱采集。通过主成分分析（PCA）和正交偏最小二乘方判别分析（OPLS-DA）筛选组间差异脂质标志物，并结合KEGG数据库对差异脂质进行代谢通路富集分析。结果：实验共鉴定出28个脂质亚类、671种脂质成分。多元统计分析显示，野生品、栽培品及混伪品间的脂质谱图具有显著的物种特异性。研究成功锁定6种仅在野生冬虫夏草中检出的特有脂质分子作为物种鉴别标志物。KEGG分析显示，差异脂质显著富集于甘油酯代谢、甘油磷脂代谢等15条关键通路。结论：野生冬虫夏草与人工栽培品及混伪品在脂质组成上存在显著差异；筛选出的6种特异性脂质标志物可作为区分野生冬虫夏草的有效分子特征。本研究为冬虫夏草的真伪精准鉴别和质量评价提供了科学依据与新的参考。

OBJECTIVE This study compares the lipid composition differences among wild Ophiocordyceps sinensis (YS), artificially cultivated varieties (RG), and common counterfeit varieties (DS, LS, and YXB), aiming to screen and identify specific lipid biomarkers that can accurately distinguish wild Ophiocordyceps sinensis. These findings provide a scientific basis for authenticating and quality evaluating Ophiocordyceps sinensis. METHODS In this study, a comprehensive non-targeted lipidomics strategy was employed. Lipid extraction from five types of Cordyceps samples was performed, followed by analysis using ultra-high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS). Multivariate statistical analyses, including Principal Component Analysis (PCA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA), were used to screen for differential lipid markers. Biological functions and metabolic pathways were further explored through KEGG pathway enrichment analysis. RESULTS A total of 671 lipid species across 28 subclasses were identified and quantified. Significant lipidomic variations were observed among wild, cultivated, and adulterant samples. Six unique lipid molecules were successfully identified as specific biomarkers for wild Ophiocordyceps sinensis. Furthermore, 15 metabolic pathways were significantly enriched, with glycerolipid metabolism and glycerophospholipid metabolism being the most prominent. CONCLUSION There are significant differences in lipid composition between wild Ophiocordyceps sinensis, artificially cultivated varieties, and adulterant samples; the six specific lipid markers selected can serve as effective molecular characteristics for distinguishing wild Ophiocordyceps sinensis. This study provides a scientific basis and new references for the accurate identification and quality evaluation of Ophiocordyceps sinensis.