首乌苷制剂中二苯乙烯苷含量测定及其在小鼠体内组织分布研究

Determination of 2,3,5,4′-tetrahydroxy-tilbene-2-O-β-D-glucoside in Shouwugan Preparation and Its Tissue Distribution in Mouse

关键词:  二苯乙烯苷  高效液相色谱法  含量测定  组织分布

Key words:  stilbene glucoside  HPLC  content determination  tissue distribution

摘要:

目的 建立首乌苷制剂中二苯乙烯苷(TSG)含量测定方法；观察其在小鼠体内的组织分布。方法 采用反相高效液相色谱法，色谱柱：Agilent XDB C₁₈(150 mm×4.6 mm，5 μm)，流动相：乙腈-水(20∶80)；检测波长：320 nm。取小鼠，灌胃给予首乌苷样品溶液60 min后放血处死，取心、肝、脾、肺、肾、脑、睾丸，制备组织匀浆，甲醇除蛋白，测定各脏器中TSG的含量。结果 TSG在0.053~0.489 μg内线性关系良好(r=0.999 3)，低、中、高浓度的回收率分别为(101.6±1.3)%，(100.9±2.3)%，(100.8±2.4)%。TSG在肝组织中分布最多，其次为肺、肾、心等。结论 TSG含量测定方法简便可靠、重复性好，可用于首乌苷制剂的质量控制。TSG在小鼠多个脏器中均有分布，以肝脏中分布最多，可通过血脑屏障。

Abstract:

OBJECTIVE To establish a method for the determination of 2,3,5,4′-tetrahydroxy-tilbene-2-O-β- D-glucoside (TSG) in Shouwugan preparation, and to study the tissue distribution of TSG in mouse. METHODS Analyses was performed at 25 ℃ on Agilent XDB C₁₈(150 mm×4.6 mm, 5 μm) column with acetonitrile-water (20∶80) as mobile phase at a flow rate of 1.0 mL·min^-1. The detection wavelength was set at 320 nm. RP-HPLC method was developed for determination of TSG in organs of mouse. RESULTS Linearity of TSG was good at 0.053-0.489 μg. The recovery of TSG at low, middle and high concentration was (101.6±1.3)%, (100.9±2.3)%, (100.8±2.4)%, respectively. The TSG was distributed on liver mostly, the following was lung, kidney, heart and so on. CONCLUSION The method is simple and accurate, which can be used for quality control of Shouwugan preparation. The TSG is distributed on many organs, especially on liver, and it can affect brain stem by passing throw blood-brain barrier.