目的 建立RP-HPLC法測(cè)定兔血漿中鹽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），流動(dòng)相為甲醇–0.5%冰乙酸（35∶65），體積流量為1.0 mL/min，檢測(cè)波長(zhǎng)280 nm，柱溫30℃，香豆素作內(nèi)標(biāo)，采用甲醇沉淀蛋白法處理血漿樣品。結(jié)果 鹽酸川芎嗪線性范圍為0.072～13.888 μg/mL（r=0.999 8），最低檢測(cè)濃度12 ng/mL，日內(nèi)、日間精密度試驗(yàn)的RSD值均小于2%；阿魏酸線性范圍為0.06～15.36 μg/mL（r=0.999 9），最低檢測(cè)濃度10 ng/mL，日內(nèi)、日間精密度試驗(yàn)的RSD值均小于3%。結(jié)論 該分析方法簡(jiǎn)便、靈敏，可用于同時(shí)測(cè)定兔血漿中鹽酸川芎嗪和阿魏酸。

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.

國(guó)家重大新藥創(chuàng)制科技重大專項(xiàng)（2011ZX09102-003-05）