RP-HPLC法测定兔血浆中盐酸川芎嗪和阿魏酸

Determination of tetramethylpyrazine hydrochloride and ferulic acid in rabbit plasma by RP-HPLC

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（250mmx4．6mm，5μm），流动相为甲醇-0．5％冰乙酸（35：65），体积流量为1．0mL／min，检测波长280nm，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0．07213．888gg／mL（r=0．9998），最低检测浓度12ng／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0．06～15．36gg／mL（F0．9999），最低检测浓度10ng／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。

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 /am） 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 /ag/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 tests were within 3%. Conelusion The method is rapid and sensitive and could be used for simultaneous determination of TMPH and ferulic acid in plasma of rabbits.