基于UPLC-Q-TOF-MS/MS技术对藏药宽筋藤中生物碱类成分的快速辨识  

Rapid identification of alkaloids component in Tibetan medicine Tinospora sinensis based on UPLC-Q-TOF-MS/MS technique

作　　者：张武岗[1] 郎一帆 姚云 杨武亮 杨世林[1] 陈海芳 冯育林[1] ZHANG Wu-gang;LANG Yi-fan;YAO Yun;YANG Wu-liang;YANG Shi-lin;CHEN Hai-fang;FENG Yu-lin(National Pharmaceutical Engineering Center(NPEC)for Solid Preparation in Chinese Herbal Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004,China;Key Laboratory of Modern Chinese Medicine Preparations,Ministry of Education,Jiangxi University of Chinese Medicine,Nanchang 330004,China)

机构地区：[1]江西中医药大学中药固体制剂制造技术国家工程研究中心,江西南昌330004 [2]江西中医药大学现代中药制剂教育部重点实验室,江西南昌330004

出　　处：《中草药》2023年第24期7977-7988,共12页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(81903917);江西省中医药管理局科技计划项目(2022Z023);江西省自然科学基金项目(20192BAB205109);江西省教育厅科学技术研究项目(GJJ2200908,GJJ170723);江西中医药大学博士科研启动基金项目(2018WBZR002);江西中医药大学科技创新团队项目(CXTD22001);江西民族传统药现在科技与产业发展协同创新中心开放基金项目(JXXT201402008);中央引导地方科技发展资金项目(20212ZDD02009)。

摘　　要：目的 采用超高效液相色谱-四级杆-飞行时间质谱(ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry,UPLC-Q-TOF-MS/MS)对藏药宽筋藤Tinospora sinensis藤茎中生物碱类成分进行分析和鉴定。方法 采用色谱柱Waters ACQUITY UPLC C_(18)色谱柱(100 mm×2.1 mm,1.7μm),以流动相乙腈-0.1%甲酸水溶液进行梯度洗脱。使用电喷雾离子源(ESI),正离子模式下采集数据;通过测定宽筋藤生物碱对脂多糖诱导小鼠巨噬RAW264.7细胞释放一氧化氮的抑制能力,评价其抗炎活性。结果 通过二级高分辨质谱分析结合对照品数据及相关文献,共鉴定出73个生物碱,包括12个原小檗碱型生物碱、23个四氢原小檗碱型生物碱、20个苄基异喹啉类生物碱、9个阿朴啡类生物碱、9个其他类生物碱。抗炎活性检测表明,宽筋藤生物碱对脂多糖刺激的RAW264.7细胞的一氧化氮具有较强的抑制作用。结论对藏药宽筋藤生物碱类成分进行较为全面系统地解析,为药效物质基础研究及开发应用奠定基础。Objective Ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry(UPLC-Q-TOF-MS/MS)method was developed to analyze and identify the alkaloids from Tibetan medicine Tinospora sinensis.Methods The analysis was performed on a Waters ACQUITY UPLC C_(18) column(100 mm×2.1 mm,1.7µm)with acetonitrile and 0.1%formic acid water solution as the mobile phase for gradient elution.The electrospray ion source(ESI)was adopted to collect the data in positive ionization mode.The anti-inflammatory effect of alkaloids from Tinospora sinensis was evaluated with the inhibition on the release of nitric oxide in RAW264.7 cells induced by lipopolysaccharide.Results A total of 73 alkaloids,including 12 protoberberine-type alkaloids,23 tetrahydroprotoberberine-type alkaloids,20 benzylisoquinoline alkaloids,9 aporphine alkaloids and 9 other alkaloids were identified by secondary high resolution mass spectrometry combined with reference substances data and related literature.The results of anti-inflammatory activity showed that the alkaloids of Tinospora sinensis showed strong inhibition effect on the release of nitric oxide in RAW264.7 cell induced by lipopolysaccharide.Conclusion The established method systematically analyzed the alkaloids of the Tibetan medicine. It provides a reference for the subsequent study of material basis and development of Tinospora sinensis.

关 键 词：UPLC-Q-TOF-MS/MS 宽筋藤 生物碱 成分分析 小檗碱 巴马汀 四氢巴马汀 黄柏碱 木兰花碱 木兰箭毒碱 

分 类 号：R284.1[医药卫生—中药学]