Chen Jun, Hou Ting, Fang Yun, Chen Zhi-peng, Liu Xiao, Cai Hao, Lu Tu-lin, Yan Guo-jun, Cai Bao-chang
College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, China.
Yakugaku Zasshi. 2011;131(5):721-9. doi: 10.1248/yakushi.131.721.
A simple and low-cost HPLC method with UV absorbance detection was developed and validated to simultaneously determine strychnine and brucine, the most abundant alkaloids in the processed Semen Strychni, in rat tissues (kidney, liver, spleen, lung, heart, stomach, small intestine, brain and plasma). The tissue samples were treated with a simple liquid-liquid extraction prior to HPLC. The LOQs were in the range of 0.039-0.050 µg/ml for different tissue or plasma samples. The extraction recoveries varied from 71.63 to 98.79%. The linear range was 0.05-2 µg/ml with correlation coefficient of over 0.991. The intra- and inter-day precision was less than 15%. Then the method was used to measure the tissue distribution of strychnine and brucine after intravenous administration of 1 mg/kg crude alkaloids fraction (CAF) extracted from the processed Semen Strychni. The results revealed that strychnine and brucine possessed similar tissue distribution characterization. The highest level was observed in kidney, while the lowest level was found in brain. It was indicated that kidney might be the primary excretion organ of prototype strychnine and brucine. It was also deduced that strychnine and brucine had difficulty in crossing the blood-brain barrier. Furthermore, no long-term accumulation of strychnine and brucine was found in rat tissues.
建立并验证了一种简单且低成本的高效液相色谱法（HPLC），该方法采用紫外吸光度检测，用于同时测定大鼠组织（肾脏、肝脏、脾脏、肺、心脏、胃、小肠、脑和血浆）中马钱子炮制后含量最高的两种生物碱士的宁和马钱子碱。在进行HPLC分析之前，组织样品采用简单的液-液萃取法进行处理。不同组织或血浆样品的定量下限（LOQ）在0.039 - 0.050 µg/ml范围内。萃取回收率在71.63%至98.79%之间。线性范围为0.05 - 2 µg/ml，相关系数超过0.991。日内和日间精密度均小于15%。然后，该方法用于测定静脉注射1 mg/kg从马钱子炮制后提取的总生物碱组分（CAF）后士的宁和马钱子碱的组织分布。结果表明，士的宁和马钱子碱具有相似的组织分布特征。在肾脏中观察到的含量最高，而在脑中含量最低。这表明肾脏可能是原型士的宁和马钱子碱的主要排泄器官。还推断出土的宁和马钱子碱难以穿过血脑屏障。此外，在大鼠组织中未发现士的宁和马钱子碱的长期蓄积。
