卵类糖蛋白固定相高效液相色谱法测定血浆中尼伐地平对映体(英文)

Determination of nilvadipine enantiomers in plasma by HPLC based on ovoglycoprotein as stationary phase

目的 建立血浆中尼伐地平对映体的HPLC法。方法在弱碱性条件下，血浆样品用环己烷-异丙醇（96．5：3．5，V/V）混合溶剂提取,以卵类糖蛋白为固定相，以含乙醇22．5％的50mmol·L^-1磷酸盐缓冲液（pH5．1）为流动相，采用HPLC法分离测定，检测波长236nm。结果在本实验条件下，血浆内源性物质无干扰，尼伐地平对映体可在30min内得到基线分离。检测信号峰面积与（S）-（＋）或（R）-（-）尼伐地平在10—300μg·L^-1内呈良好的线性关系，检测限约为3μg·L^-1，方法回收率为91．7％～100．5％，RSD〈9．8％。结论本方法具有简便快速、精密度和准确度好的优点，可用于血浆中尼伐地平对映体的分离和测定。

AIM To establish a HPLC method for the separation and determination of nilvadipine enantiomer in plasma. METHODS Plasma samples were extracted with a mixed solvent of cyclohexane-isopropanol （96.5：3.5, V/V） under weak alkaline condition. Then the separation was performed in a column packed with ovoglycoprotein as the stationary phase and 50 mmool·L^-1 sodium phosphate buffer （pH 5.1 ）, containing 22.5 % ethanol, as the mobile phase. The column effluent was monitored by UV detection and the wavelength of 236 nm was selected. RESULTS Under experimental conditions, no endogenous interference was observed, and the separation and determination of nilvadipine enantiomer in plasma were achieved within 30 rain. The detector response （peak area） was linear in the range of 10 - 300 μg.L^-l for （S） - （ ＋ ） - or （R） - （ - ）-nilvadipine, with the detection limits around 3 μg. L^-1. The recoveries were in the range of 91.7 % - 100.5 %, with relative standard deviation （RSD） less than 9.8 %. CONCLUSION The suggested procedure was validated and found to be rapid, precise and accurate for the separation and determination of nilvadipine enantiomer in plasma.