[关键词]
[摘要]
目的 建立RP-HPLC法测定兔血浆中盐酸川芎嗪和阿魏酸的方法。方法 采用RP-HPLC法,使用Diamonsil C18柱(250 mm×4.6 mm,5 μm),流动相为甲醇–0.5%冰乙酸(35∶65),体积流量为1.0 mL/min,检测波长280 nm,柱温30℃,香豆素作内标,采用甲醇沉淀蛋白法处理血浆样品。结果 盐酸川芎嗪线性范围为0.072~13.888 μg/mL(r=0.999 8),最低检测浓度12 ng/mL,日内、日间精密度试验的RSD值均小于2%;阿魏酸线性范围为0.06~15.36 μg/mL(r=0.999 9),最低检测浓度10 ng/mL,日内、日间精密度试验的RSD值均小于3%。结论 该分析方法简便、灵敏,可用于同时测定兔血浆中盐酸川芎嗪和阿魏酸。
[Key word]
[Abstract]
Objective To establish a RP-HPLC method for simultaneous determination of tetramethylpyrazine hydrochloride (TMPH) and ferulic acid in plasma of rabbits. Methods Separation was carried out on a Diamonsil C18 reverse phase column (250 mm × 4.6 mm, 5 μm) at a flow rate of 1.0 mL/min. Chromatography was performed at 30 ℃. TMPH and ferulic acid were monitored at a wavelength of 280 nm with coumarin as internal standard, and injection volume was 20 μL. Methanol - 0.5% acetic acid solution (35∶65) was used as mobile phase. Results The calibration curves for TMPH in plasma were linear in the range of 0.072 — 13.888 μg/mL (r=0.999 8), and the detection limit of TMPH in plasma was 12 ng/mL. RSD values of the intra-day and inter-day precision tests were within 2%. The calibration curves for ferulic acid in plasma were linear in the range of 0.06 — 15.36 μg/mL (r=0.999 9), and the detection limit of ferulic acid in plasma was 10 ng/mL. RSD values of the intra-day and inter-day precision testswere within 3%. Conclusion The method is rapid and sensitive and could be used for simultaneous determination of TMPH and ferulic acid in plasmaof rabbits.
[中图分类号]
[基金项目]
国家重大新药创制科技重大专项(2011ZX09102-003-05)