高分离快速液相色谱-四极杆飞行时间质谱法检测人参皂苷Re在大鼠体内的分布

Tissue Distribution of Ginsenoside Re in Rats by Rapid Resolution Liquid Chromatography Coupled With Quadrupole-Time-of Flight Mass Spectrometry

建立了高分离快速液相色谱-四极杆飞行时间质谱（RRLC-Q-TOF MS）法检测大鼠尾静脉注射人参皂苷Re（G-Re）后,不同组织（心、肝、脾、肺、肾、脑）中G-Re的分布情况。样本经蛋白沉淀处理,Supelco AscentisExpress C18色谱柱（50mm×3.0mm×2.7μm）分离,采用含0.1%甲酸的水溶液（A）-乙腈（B）为流动相,梯度洗脱,以人参皂苷Rc（G-Rc）作为内标,在电喷雾负离子模式下检测。结果表明：组织匀浆标准曲线的线性范围为10-20 000μg/L,线性关系良好（r〉0.99）,最低检出限（S/N=3）为3μg/L,最小定量限为10μg/L;准确度、日内和日间精密度、提取回收率均符合生物样品的分析要求。大鼠尾静脉注射20mg/kg G-Re溶液后,在大鼠体内不同组织中均能检测到G-Re,主要分布在肺、脾、肾、心、脑、肝等器官,其中在肺中分布最多,在肝中分布最少。该方法简单、灵敏、快速、检出限低,适用于检测人参皂苷Re在生物体内的分布。

A method of rapid resolution liquid chromatography coupled with quadrupole- time-of flight mass spectrometry （RRLC-Q-TOF MS） was established to investigate the distribution of ginsenoside Re （G-Re） in rat various tissues （heart, liver, spleen, lung,kidney and brarin） after a single administration by intravenous injection. The biological samples were prepared by protein precipitation. Chromatographic separations was achieved on a Supelco Ascentis Express C18 column （50 mm×3. 0 mm×2. 7 jam） with a mobile phase consisting of solvent A （0. 1 % formic acid in water ） and solvent B （acetonitrile） at a flow rate of 0. 3 mL/min. Gradient elution was as follows： 0-15 m in , 20%-40%B; 15-20 min, 4 0%-5 0%B ; 20-22 min, 5 0% -1 0 0% B . Ginsenoside Rc （G -Rc ） was used as internal standard （IS）. All mass spectrometric experiments were performed on RRLC-Q-TOF MS equipped with an electrospray ionization （ESI） source operated in negative mode. The result shows that the calibration curve is linear in the range of 10-20 000 μg/L for tissue homogenates （r〉0. 99）,the lower limit of detection （LLOD） is 3 μg/L, and the lower limit of quantification （LLOQ） is 10μg/L. The accuracy, intra-day, inter-day precision and recovery ratio can meet the requirements of the biologi-cal samples analysis. As a result, G-Re could be widely distributed in organizations with 20 mg/kg solution by caudal vein injection, which could be detected in different organs and distributed mainly in the lung, spleen,kidney and heart. Lung had a highest affinity to G-Re, which was not easy to pass through the blood-brain barrier and hard to accumulate. This method is simple,sensitive and rapid, which is suitable for analyzing ginsenosides Re in vivo.