HILIC/ESI-Orbitrap HRMS法结合Compound Discoverer软件快速智能化鉴定瓜蒌皮注射液中的化学成分

Rapid and intelligentized identification of the constituents in Gualoupi injection by HILIC/ESI-Orbitrap HRMS combined with Compound Discoverer software

本研究采用亲水作用色谱柱串联四极杆-静电场轨道阱-线性离子阱三合一组合式高分辨质谱仪(HILIC/Orbitrap Fusion Lumos HRMS)建立了快速智能化鉴定瓜蒌皮注射液中化学成分的分析方法。采用Compound Discoverer软件,检索在线数据库ChemSpider和mzCloud,共鉴定了48个化学成分,包括生物碱、氨基酸、核苷及其碱基等,其中25个通过与对照品对照进行了准确鉴定,基本阐明了瓜蒌皮注射液的化学物质基础。采用该方法采集21批样品数据,以48个成分的峰面积为观察值,进行主成分分析,考察不同批次样品间的稳定性,结果显示注射液批间差异性的主要来源为原料药材,注射液制备工艺相对稳定。本研究为瓜蒌皮注射液的药效物质基础研究及质量标准提高研究提供了重要参考。

A rapid identification of the constituents in Gualoupi injection was developed by HILIC/Orbitrap Fusion Lumos HRMS. ACQUITY XBridge Amide column was used to isolate the constituents with large polarity. ESI with positive ion mode was employed and ＂Top Speed＂ DDA was applied in the MS^2 scan. Compound identification was carried out by using Compound Discoverer software through comparing the precursor and product ions information with those in ChemSpider and mzCloud database. As a result, 48 compounds was identified, including alkaloids, amino acid, nucleosides, nucleobases, etc., among which 25 was unambiguously identified by comparing the retention time and mass spectra information with those of reference standards. In general, the main constituents from Gualoupi injection were explained in this study. Additionally, full-scan mass spectra of 21 batches of the injection were collected by the established method. Subsequently, principal component analysis（PCA） with the peak area as the observation ID was employed to analyze the discrimination of different bathes of samples. The results indicated that the batch-to-batch difference was generally from the crude drug, and the preparation process of this injection was relatively stable. In conclusion, a rapid and effective method for the identification of compounds in Gualoupi injection was established by using UHPLC-Orbitrap Fusion Lumos HRMS and the inter-bath stability was also investigated, which may make a great contribution to the study of the bioactive ingredients in Gualoupi injection and also provide vital evidence for the standard promotion.