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

摘要

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 μm)with a mobile phase consisting of solvent A (0.1% formic acid in water)and solvent B (ace-tonitrile)at a flow rate of 0.3 mL/min.Gradient elution was as follows:0-15 min, 20%-40%B;15-20 min,40%-50%B;20-22 min,50%-100%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 10 μg/L.The accuracy,in-tra-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.%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(50 mm×3.0 mm×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10~20000μg/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20 mg/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。