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目的:建立测定大鼠大血管中异钩藤碱含量的方法。方法:Wista大鼠灌胃钩藤水煎液后取出大血管,液氮研磨,用乙醇沉淀组织匀浆中的蛋白质,取上清液用氮气吹干,加甲醇复溶后取上清液直接进样,用高效液相色谱仪测定。RP18WatersTM色谱柱(4.6 mm×250 mm,5μm),流动相甲醇-水(70∶30),三乙胺调pH 8.0,流速0.8 mL·min-1,检测波长254 nm,异钩藤碱保留时间约为12 min,柱温30℃。结果:大血管中异钩藤碱的质量浓度在0.04~20.0 mg·L-1,大鼠峰面积之比Y与异钩藤碱的质量浓度X具有较好的线性关系,回归方程为Y=4.644 1X+0.161 2(R2=0.999 2);平均提取回收率为99.34%,RSD<8.0%。结论:方法简便、稳定、易行,可以用于测定大鼠大血管中异钩藤碱的含量。
Objective: To establish a method for the determination of isorhline in rats’ large blood vessels. Methods: The Wistar rats were gavaged with Uncaria rhynchophylla decoction and the blood vessels were removed. The blood was ground with nitrogen, and the protein in the tissue homogenate was precipitated with ethanol. The supernatant was blown dry with nitrogen. The supernatant Injection, measured by high performance liquid chromatography. The mobile phase consisted of methanol-water (70:30), RP18WatersTM (4.6 mm × 250 mm, 5 μm), triethylamine adjusted to pH 8.0 and the flow rate was 0.8 mL · min-1. The detection wavelength was 254 nm. About 12 min, column temperature 30 ℃. Results: The concentration of isorhline in large blood vessels was between 0.04 and 20.0 mg · L-1. The linear relationship between the peak area ratio (Y) of rat and the concentration of iso-rhynchophylline had a good linearity. The regression equation was Y = 4.644 1X + 0.161 2 (R2 = 0.999 2). The average extraction recovery was 99.34% with RSD <8.0%. Conclusion: The method is simple, stable and easy to determine, and can be used to determine the content of isorhline in rats’ large blood vessels.