目的:建立测定新西兰兔血浆中鹿角缩酮B的反相高效液相色谱方法,并测定静脉给药后新西兰兔血浆中鹿角缩酮B浓度及药动学参数。方法:血浆样品经酸化后用乙酸乙酯进行液液萃取,化合物1为内标,色谱柱为Phenomenex C18,流动相为甲醇-乙腈-水(30∶30∶40),流速0.8 mL.min-1,检测波长220 nm。结果:本方法中内源性物质不干扰测定,鹿角缩酮B在0.4~50μg.mL-1范围内线性良好(r=0.9995),检测限为0.2μg.mL-1;高中低浓度平均回收率为85.2%~105.5%,日内和日间精密度RSD均小于10%。结论:首次建立了鹿角缩酮B在新西兰兔体内的分析方法,为鹿角缩酮B的体内分析奠定了方法学基础。本方法专属、准确、灵敏,适用于测定新西兰兔的血药浓度;静脉给药后鹿角缩酮B在体内药代动力学符合二室模型。
Objective:To develop a reverse phase HPLC method for the determination of xyloketal B in rabbit plasma and its pharmacokinetics study.Methods:Plasma samples were extracted by liquid-liquid extraction with ethyl acetate.For quantification,compound 1 was chosen as an internal standard.The separation was performed on a Phenomenex C18 column with an isocratic mobile phase of methanol-acetonitrile-water(30∶ 30∶ 40) at a flow rate of 0.8 mL·min-1.The eluent was monitored at 220nm.Results:The method showed a good speicificity between analytes and matrix.A good linearity was observed over the concentration range of 0.4-50 μg·mL-1 with correlation coefficients of 0.9995.The limits of detection(LOD) was 0.2 μg·mL-1.The mean recovery of xyloketal B in QC samples at concentration of 0.8,10 and 40 μg·mL-1 were between 85.2%~105.5%,intra-and inter-day precision RSDs were all less than 10%(n=5).Conclusions:The analysis method of xyloketal B in rabbit plasma was established for the first time,which lay a methodological foundation for the biopharmaceutical analysis of xyloketal B.The developed method was specific,accurate and sensitive.The results indicated that the analytic method was satisfactorily applied to perform preclinical pharmacokinetic study of xyloketal B in rabbit plasma.The plasma concentration-time curve fitted the two-compartment model after intravenous administration in rabbits.