高效液相荧光色谱法测定人血浆中细辛脑的浓度

Determination of Asarone in human plasma by high performance liquid chromatography with fluorescence

目的:建立测定人血浆中细辛脑浓度的高效液相荧光色谱法。方法:血浆样品经10%高氯酸沉淀后,以甲醇-0.5%醋酸液(78:22)为流动相,流速为1 ml·min^(-1),色谱柱为BaiAn-C_(18)(150 mm×4.6 mm,5μm),柱温为35℃,荧光检测波长:激发波长(λex)265 nm,发射波长(λem)395 nm。结果:血浆内源性杂质不干扰待测物测定,细辛脑的线性范围为1～300μg·L^(-1),定量下限为1μg·L^(-1),日内、日间RSD均小于5%。样品3次冻融及在沉淀后,4℃下12 h内稳定性良好。结论:该法灵敏、快速、准确,操作简便,线性范围宽,可用于细辛脑的临床药动学研究。

Objective： To establish a high performance liquid chromatography with fluorescence method for determination of Asarone in human plasma. Method： Plasma samples were treated by perchloric acid （ 10% ）. The BaiAn-C18 column （ 150 mm × 4.6mm, 5 μm） at 35 ℃ was used. The mobile phase consisted of methanol：0.5% acetic acid （78： 22） pumped at a flow rate of 1 ml·min^-1. The fluorescence detector was operated at,an excitation and emission wavelengths 265 nm and 395 nm, respectively. Result： The intrinsic impurity of plasma did not interfere with the determination of analyte. The linear range of Asarone was 1 - 300 μg· L^-1. Lower limit of quantification was 1 μg· L^-1. The precision of inter-day were less than 5%. The stability of unprocessed and processed samples stored at 4 ℃ for 12 hours was good. After 3 cycles of freeze and thaw processes,the plasma samples were stable. Conclusion： The established HPLC-FLU method is fast, sensitive, and accurate and can be applied to the clinical pharmacokinetics investigations of Asarone.