苗药红禾麻提取物在离体人肠道菌群中的代谢研究
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篇名: 苗药红禾麻提取物在离体人肠道菌群中的代谢研究
TITLE: Study on Metabolism of Miao Medicine Laportea bulbifera Extract in Isolated Human Intestinal Flora
摘要: 目的:探索苗药红禾麻提取物在离体人肠道菌群中的代谢特征。方法:取红禾麻药材,以70%乙醇回流提取,经浓缩、正丁醇萃取、干燥后,得红禾麻提取物。将0.05g/mL红禾麻提取物溶液1mL与离体人肠道菌液10mL混合后,在厌氧环境下共同培养36h(同时设置不含药物或人肠道菌液的空白对照),模拟该提取物在人体肠道内的代谢过程。采用超高效液相色谱-四极杆-飞行时间质谱联用技术(UPLC-Q-TOF/MS)对厌氧反应后的代谢产物进行检测。色谱柱为AgilentEclipsePlusC18RRHD,流动相为0.01%甲酸水溶液-0.01%甲酸乙腈溶液(梯度洗脱),柱温为40℃,流速为0.25mL/min,进样量为1µL;采用电喷雾离子源(ESI),以负离子模式(ESI-)扫描,毛细管电压为4.5kV,离子源温度为120℃,碰撞能为15~32V,扫描范围为m/z50~1000。采用MassLynxV4.1软件中的“Strip”模块分析获得红禾麻提取物反应液与空白对照的差异图谱。根据质谱数据和UNIFI软件进行代谢产物的相对分子量和分子式预测,结合对照品信息和相关文献报道,推测红禾麻提取物在离体人肠道菌群中的代谢产物结构及其可能的生物转化途径。结果与结论:红禾麻提取物经离体人肠道菌群代谢后,共检出3个原型产物(芦丁、槲皮苷、山柰酚-3-O-芸香糖苷)和22个代谢产物(主要为槲皮素、单咖啡酰基奎宁酸、异槲皮苷等的代谢产物),其主要生物转化途径是以还原、氧化、水解为主的Ⅰ相反应。
ABSTRACT: OBJECTIVE:To explore the metabolic charact eristics of Miao medicine Laportea bulbifera extract in isolated human intestinal flora. METHODS :L. bulbifera was extracted with 70% ethanol reflux extraction. After concentration,extraction with n-butanol and drying ,L. bulbifera extract was obtained. Taking 0.05 g/mL L. bulbifera extract 1 mL mixed with isolated human intestinal flora fluid 10 mL and cultured for 36 h in anaerobic environment (setting up blank control without drugs or human intestinal bacterial solution ),so as to simulate the metabolic process of the extract in human intestine. The metabolites were detected by UPLC-Q-TOF/MS. The determination was performed on Agilent Eclipse Plus C 18 RRHD column with mobile phase consisted of 0.01% formic acid water solution- 0.01% formic acid acetonitrile solution (gradient eluetion )at the flow rate of 0.25 mL/min. The column temperature was set at 40 ℃,and the sample size was 1 µL. ESI detection was adopted and scanned by negative ion mode (ESI-);the capillary voltage was 4.5 kV,the ion source temperature was 120 ℃,the collision energy was 15-32 V,and the scanning range was m/z 50-1 000. The “Strip”module of MassLynx V 4.1 software was used to analyze the differential chromatograms between the reaction solution and the blank control of L. bulbifera extract. Mass spectrum data and UNIFI so ftware were used to predict relative molecular weight and formula ;based on the information of substance control and related literature reports , the structure and biotransformation pathway of L. bulbifera metabolites in isolated human intestinal flora were predicted and analyzed. RESULTS & CONCLUSIONS : A total of 3 prototype : products(rutin,quercetin,kaempferol-3-O-rutinoside)and 22metabolites (mainly the metabolites of quercetin ,mono- caffeoylquinic acid ,isoquercitrin,etc.) were detected after metabolized in isolated human intestinal flora. Itsbiotransformation pathway is phase Ⅰ reaction,which mainly consisted of reduction ,oxidation and hydrolysis.
期刊: 2020年第31卷第14期
作者: 薛寸,吴耽,巩仔鹏,陈思颖,唐娟,李月婷,王爱民,李勇军,兰燕宇,王永林
AUTHORS: XUE Cun,WU Dan,GONG Zipeng,CHEN Siying,TANG Juan,LI Yueting,WANG Aimin,LI Yongjun,LAN Yanyu,WANG Yonglin
关键字: 红禾麻;离体人肠道菌群;代谢;超高效液相色谱-四极杆-飞行时间质谱联用技术;鉴定
KEYWORDS: Laportea bulbifera ;Isolated human intestinal flora ;Metabolism;UPLC-Q-TOF/MS;Identification
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