胆类中药体内代谢产物甘氨胆酸硫酸盐同分异构体的区分  

Differentiation of Isomers of Glycine-conjugated Bile Acid Sulfates——In vivo Metabolites of Animal Gallbladderderived Chinese Medicine

作　　者：张珂 李玮 王艳敏 龚兴成 曹丽波 宋月林 宋青青[1] 屠鹏飞[1] ZHANG Ke;LI Wei;WANG Yan-min;GONG Xing-cheng;CAO Li-bo;SONG Yue-lin;SONG Qing-qing;TU Peng-fei(Modern Research Center for Traditional Chinese Medicine,School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学中药学院中药现代研究中心,北京100029

出　　处：《中国现代中药》2021年第11期1883-1888,共6页Modern Chinese Medicine

基　　金：国家自然科学基金项目(81973444,82003911,81773875)。

摘　　要：目的:通过在线能量分辨质谱法(Online ER-MS)快速区分胆类中药在体内的代谢产物甘氨胆酸硫酸盐同分异构体。方法:采用高效液相色谱-质谱法(HPLC-MS)采集3个甘氨胆酸硫酸盐异构体甘氨熊去氧胆酸-3-磺酸盐(GUDCA-3-S)、甘氨鹅去氧胆酸-3-磺酸盐(GCDCA-3-S)、甘氨去氧胆酸-3-磺酸盐(GDCA-3-S)的高分辨多级质谱信息,推导质谱裂解途径并归属各碎片离子,构建候选离子对m/z 528.3→528.3、528.3→448.3、528.3→404.3、528.3→386.3、528.3→330.2、528.3→97.0和528.3→74.0,每个候选离子对进一步衍生1组拟离子对(PITs),对应1组在–5~–180 eV梯度变化(步长为1 eV)的碰撞能;通过Online ER-MS采集所有拟离子对质谱响应,采用高斯曲线拟合每一组拟离子对的响应值,构建裂解曲线组,进而在同分异构体间比较同一离子对的裂解曲线。结果:3个同分异构体的MS1和MS2基本一致,难以区分。各组拟离子对的响应值均呈现高斯分布。同一化合物的不同候选离子对呈现出不同的裂解曲线,同分异构体的相同离子对的裂解曲线也存在差异。结论:利用Online ER-MS可以实现甘氨胆酸硫酸盐同分异构体的快速、有效区分,为胆类中药体内药效物质和作用机制的阐明提供参考。Objective:To differentiate glycine-conjugated bile acid sulfate isomers,the in vivo metabolites of animal gallbladder-derived Chinese medicine,by online energy-resolved mass spectrometry(Online ER-MS).Methods:highperformance liquid chromatography-mass spectrometry was employed to acquire the accurate m/z values of precursor and product ions of three glycine bile acid sulfates isomers,namely,glycoursodeoxycholic acid-3-sulfate,glycochenodeoxycholic acid-3-sulfate,and glycodeoxycholic acid-3-sulfate,and then,the mass fragmentation patterns were analyzed,accompanied with ion assignment.Seven ion transition candidates were constructed as m/z 528.3→528.3,528.3→448.3,528.3→404.3,528.3→386.3,528.3→330.2,528.3→97.0,and 528.3→74.0,and each afterwards derived a set of pseudo-ion transitions(PITs),corresponding to a series of progressive collision energies from–5 to–180 eV(step size:1 eV).Mass responses of all PITs were recorded via online ER-MS,and corresponding breakdown graph sets were developed by Gaussian curve fitting.The breakdown graph of the same ion pair was compared between isomers.Results:The MS1 and MS2 data of three isomers were basically the same,which failed to differentiate them.The mass response of each set of PITs took on Gaussian distribution.Different ion transitions of the same compound showed different breakdown graphs,so did the same ion pairs of isomers.Conclusion:Online ER-MS is a promising method to facilitate rapid and efficient isomer identification,and provides reliable information for clarifying the pharmacodynamic substances and mechanism of animal gallbladder-derived Chinese medicine.

关 键 词：胆类中药 在线能量分辨质谱 甘氨胆酸硫酸盐 同分异构体 裂解曲线组 

分 类 号：R284[医药卫生—中药学]