基于液相色谱-高分辨质谱技术和多重质量亏损过滤技术的大鼠血浆中大黄素代谢产物鉴定

Identification of Emodin Metabolites in vivo by HPLC-HRMS and MMDF Technique

目的:利用高效液相色谱-高分辨质谱法(HPLC-HRMS)和多重质量亏损过滤法(MMDF)识别和鉴定大鼠血浆中大黄素的代谢产物。方法:利用HPLC-HRMS技术自动采集生物样品中化合物的高分辨质谱数据集;采用高分辨提取离子流(HREIC)提取可能的大黄素可预知代谢产物;根据可预知代谢产物的结果,采用MMDF将体内高暴露量的4个可预知代谢物(M9、M11、M15和M18)与大黄素作为模板化合物,采用窄范围过滤(质量数:±100,质量亏损过滤范围:±0.025),进行不可预知的代谢产物分析。结果:快速识别并鉴定了大黄素的18个代谢物,其中3个(M4、M13和M16)为首次发现。与传统的质量亏损过滤(MDF)技术相比,该方法能够更显著地降低假阳性结果。结论:建立了一种简便、可靠、高效率的大黄素体内代谢物鉴定方法,证实大黄素主要代谢途径为葡萄糖醛酸化、磺酸化等结合反应,为大黄素体内研究提供参考。

Objective:To discover and identify the metabolites of emodin in rat plasma by using high performance liquid chromatography-high resolution mass spectrometer(HPLC-HRMS)and multiple mass defect filter(MMDF)technology.Methods:HRMS datasets of compounds in the plasma samples were automatically collected by using HPLC-HRMS technology.High resolution-extracted ion chromatography(HREIC)was adopted to extract the possible predictable metabolites of emodin.According to the results of the predictable metabolites,MMDF technology was then used to conduct narrow-range filtering(mass number range:±100;mass defect filtering range:±0.025)by taking the four predictable metabolites with high exposure in vivo(M9,M11,M15 and M18)and emodin as template compounds,and unpredictable metabolites were analyzed.Results:18 metabolites were recognized and identified quickly,and 3 of them(M4,M13 and M16)were first found.Compared to traditional MDF technology,this method can cut down false positive results more significantly.Conclusion:In this study,a simple,reliable and efficient method for identifying in vivo metabolites of emodin was developed,and it was confirmed that the major metabolic pathways of emodin are glucuronic acid conjugation and sulfation,thus providing a basis for future studies on emodin.In addition,this method has a great potential in the analysis of in vivo metabolites of traditional Chinese medicines.