LC-MS/MS测定大鼠血浆中的野漆树苷及其药代动力学研究

Determination of rhoifolin in rat plasma by LC-MS/MS and its application to a pharmacokinetic study

目的建立大鼠血浆中野漆树苷的LC-MS/MS测定方法,并研究静脉注射后野漆树苷在大鼠体内的药代动力学特征。方法大鼠血浆样品经乙酸乙酯萃取处理,用色谱柱Phenomenex C8（30 mm×2.00 mm,3μm）进行分离,以甲醇-水为流动相梯度洗脱,采用电喷雾离子源（ESI）,负离子模式,多反应监测（MRM）,选择离子分别为野漆树苷（m/z 577.6→269.1）,内标柚皮素（m/z 271.0→151.0）,将建立的方法应用于测定大鼠静脉注射野漆树苷后血浆的药物浓度,并采用DAS 2.0计算得主要药代动力学参数。结果大鼠血浆中野漆树苷在1~2000μg/L浓度范围内线性关系良好,日内、日间精密度（RSD）均〈10%,准确度（RE）在±7%之间,低、中、高3个浓度下提取回收率为86.8%~91.0%,无明显基质效应且稳定性良好。大鼠静注该药（5 mg/kg）后AUC0-t为（72627.8±18067.9）μg·min/L,t1/2为（52.1±14.3）min,CLz为（0.07±0.02）L/（min·kg）。结论该检测方法简单、快速、灵敏、准确,适用于野漆树苷血药浓度监测及其药动学研究。药代动力学参数表明大鼠静注该药后体内消除较快。

Objective To develop and validate a LC-MS/MS method for determination of rhoifolin in rat plasma and investigate its pharmacokinetic properties after intravenous administration to rats. Methods The analyte was isolated from rat plasma by liquid-liquid extraction with ethyl acetate. Separation was performed on a Phenomenex C8column（30 mm×2.00 mm,3 μm）with gradient elution using water-methanol as mobile phase. Electrospray ionization（ESI）was adopted in the negative ion mode for multiple reaction monitoring（MRM）. The mass transitions were m/z 577.6→269.1 for rhoifolin,and m/z 271.0→151.0 for naringenin（internal standard）,respectively. Results The method showed good linearity over the range of 1-2000 μg/L. Intra and inter-day precisions were both less than 10%,the relative error was within ± 7%. The extraction recoveries of three concentrations（low,middle and high）ranged from 86.8% to 91.0%. Main pharmacokinetic parameters were calculated by DAS 2.0. AUC0-twas（72627.8± 18067.9）μg·min/L,t1/2was（52.1±14.3）min and CLzwas（0.07±0.02）L/（min·kg）. Conclusion The established LC-MS/MS method is specific and sensitive,and can be applied to pharmacokinetic study of rhoifolin. The pharmacokinetic parameters show that rhoifolin is eliminated rapidly in rats.