反相HPLC法测定大鼠组织中异钩藤碱含量

目的建立测定大鼠组织中异钩藤碱的方法。方法应用甲醇沉淀蛋白,然后氮吹仪吹干甲醇复溶后用反相HPLC法直接测定。以WatersTM RP18色谱柱(4.6mm×250mm,5μm)为固定相,前置保护柱为WondasilC18(4.0mm×10mm,5μm),甲醇-水(70∶30)含0.03%的三乙胺为流动相,流速为0.8mL/min,检测波长为254nm。卡马西平做内标。结果异钩藤碱在大鼠组织中的含量标准曲线方程为:Y=48.558X+0.2076(n=6),r=0.9996,在0.16～16.0μg/mL范围内线性关系良好,平均加样回收率为99.66%,RSD=1.56%。结论该方法灵敏、快速、简便,专一性和重复性好。

Determination of the Concentration of Isorhynchophylline in Tissue of Rats by RP-HPLC

Objective To establish a pretreatment mthod of the tissue sample of rat and a determination method of the concentration of isorhynchophylline in tissue of rats by RP-HPLC. Method The tissue sample were treated by adding methanol, and the supernatant was dried by nitrogen gas, the residue was redissolved in methanol, then detected by RP-HPLC directly. Waters？ RP18 （4.6 mm×250 mm, 5 μm） and pre-column Wondasil C18 （4.0 mm×10 mm, 5 μm） were used as the stationary phase. Methanol-water （70∶30） added 0.03% triethylamine was taken as mobile phase with flow rate of 0.8 mL/min. The wavelength of detection was 254 nm. Results The standard curve of Isorhynchophylline concentraton in rat tissues：Y=48.558X＋0.207 6 （n=6）, r=0.999 6, it appears a good linear relationship within the range of 0.16～16.0 μg/mL. The average recovery of was 99.66%, RSD was 1.56%. Conclusion This mehod is sensitive, rapid, simple, specific and reproducible.