基于UPLC-Q-TOF-MS/MS技术的甘草大鼠体内入血成分分析

Analysis of absorbed components in rat serum after oral administration of Glycyrrhizae Radix et Rhizoma by UPLC-Q-TOF-MS/MS

目的:采用超高效液相色谱-四级杆-飞行时间质谱(ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry, UPLC-Q-TOF-MS/MS)法分析灌胃给药后甘草提取液在大鼠体内的入血成分。方法:大鼠灌胃给予甘草提取液后,腹主动脉取血,分离血清。采用UPLC BEH C色谱柱(2.1 mm×100 mm, 1.7μm)分析,0.1%甲酸水-0.1%甲酸乙腈为流动相进行梯度洗脱。采用电喷雾离子源,正、负离子模式下对甘草提取液、给药血清和空白血清进行数据采集。结果:根据色谱峰保留时间及质谱碎片裂解规律,结合文献相关信息,共鉴定出13个入血原型成分,依次为Tachiogroside B、夏佛托苷、7,4′-二羟基黄酮、6″-O-乙酰甘草苷、芒柄花苷、甘草查尔酮B、金圣草(黄)素、异甘草素、芒柄花黄素、皂皮酸、黄宝石羽扇豆素、甘草酸、半甘草异黄酮B,主要为黄酮类、有机酸类、苷类和香豆素类。结论:该研究通过UPLC-Q-TOF-MS/MS分析技术,明确了甘草提取物中含有的13种入血原型成分,为其药效物质基础的研究提供了科学依据。

OBJECTIVE To analyze the blood components of Glycyrrhizae Radix et Rhizoma extract in rats by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS).METHODS Rats were given liquorice extract by intragastric administration, and blood was taken from abdominal aorta to separate the serum.The analysis was performed on a UPLC BEH Ccolumn(2.1 mm×100 mm, 1.7 μm)with 0.1% formic acid water and 0.1% formic acid acetonitrile as mobile phases.The data of glycyrrhiza extract, drug serum and blank serum were collected in positive and negative ionic modes using electrospray ion source.RESULTS According to the retention time of chromatographic peak, the fragmentation rule of mass spectrometry and related literature information, 13 prototype components were identified into the blood as Tachiogroside B,Schaftoside, 7,4′-Dihydroxyflavone, 6″-O-Acetylliquiritin, Ononin, Licochalcone B,Chrysoeriol, Isoliquiritigenin, Formononetin, Quillaic acid, Topazolin, Glycyrrhizic acid, Semilicoisoflavone B,mainly flavonoids, organic acids, glycosides and coumarins.Mainly flavonoids, organic acids, glycosides and coumarins.CONCLUSION In this study, UPLC-Q-TOF-MS/MS analysis technology was used to identify 13 kinds of prototype components in glycyrrhiza glycyrrhiza extract, which provided a scientific basis for the study of its pharmacodynamic substance basis.