Yue Peng, Tao Tao, Zhao Yan, Ren Jinfeng, Chai Xuyu
Division of Pharmaceutics, Shanghai Institute of Pharmaceutical Industry, ZhongShanBeiYi Road 1111, Shanghai 200437, China.
J Pharm Biomed Anal. 2007 May 9;44(1):309-12. doi: 10.1016/j.jpba.2007.02.019. Epub 2007 Feb 23.
A simple reversed-phase high-performance liquid chromatography (HPLC)-fluorescence method for the determination of Huperzine A in rat plasma was developed and validated. Separation was achieved on Kromasil C(8) column (5 microm, 150 mm x 4.6mm i.d.). The mobile phase, methanol-water-triethanol amine (45:55:0.05, v/v/v), was delivered at a flow rate of 1.0 ml/min. The eluent was monitored by a fluorescence detector with excitation wavelength at 310 nm and emission wavelength at 370 nm. No interfering peaks were observed in rat blank plasma. The relationship between Huperzine A concentration and peak-area ratio of Huperzine A to the IS was linear over the range of 2.5-250 ng/ml. The intra- and inter-day coefficients of variation were <or=10.1% and <or=14.1%, respectively. The method and extraction recovery of Huperzine A were 101.9-108.9% and 73.5-84.6%, respectively. Huperzine A was found to be stable for at least 5h at RT and 1 week at -20 degrees C. The method was applied to a pharmacokinetic study of Huperzine A in rats following intranasal administration.
建立并验证了一种简单的反相高效液相色谱（HPLC）-荧光法用于测定大鼠血浆中石杉碱甲。采用Kromasil C(8)柱（5μm，150mm×4.6mm内径）进行分离。流动相为甲醇-水-三乙醇胺（45:55:0.05，v/v/v），流速为1.0ml/min。用荧光检测器监测洗脱液，激发波长为310nm，发射波长为370nm。在大鼠空白血浆中未观察到干扰峰。石杉碱甲浓度与石杉碱甲与内标峰面积比之间在2.5-250ng/ml范围内呈线性关系。日内和日间变异系数分别≤10.1%和≤14.1%。石杉碱甲的方法回收率和提取回收率分别为101.9-108.9%和73.5-84.6%。发现石杉碱甲在室温下至少稳定保存5小时，在-20℃下稳定保存1周。该方法应用于大鼠鼻腔给药后石杉碱甲的药代动力学研究。
