目的:建立大鼠血浆中水黄皮素的定量分析方法,并对其药代动力学进行研究.方法:通过灌胃给予SD大鼠水黄皮素(10mg·kg-1),血浆样品经蛋白沉淀法处理后进行HPLC-MS/MS定量分析,以五味子醇甲为内标,采用Agilent Poroshell 120 EC-C18(50 mm × 3.0 mm,2.7 μm)色谱柱,乙腈(A)-0.1%甲酸溶液(B)为流动相,流速0.3 mL·min-1,梯度洗脱;采用电喷雾离子源(ESI),正离子多反应监测模式(MRM)扫描,水黄皮素[M+H]+m/z 293.1→277.0,五味子醇甲[M+H]+m/z 433.2→384.2;使用DAS软件计算相关药代动力学参数.结果:在1.5625~1600 ng·mL-1浓度范围内,水黄皮素线性关系良好(r=0.9997),定量下限为1.5625 ng·mL-1,精密度RSD和准确度RE均小于20%;低、中、高质控样品精密度RSD和准确度RE均小于15%,提取回收率在91.70%~100.28%,基质效应在88.35%~97.55%,稳定性RSD均小于15%.水黄皮素在雌性SD大鼠体内Tmax、t1/2、Cmax和AUC0→t分别为3h、1.95 h、1214.85 ng·mL-1 和 11920.43 ng·mL-1·h;在雄性SD大鼠体内Tmax、t1/2、Cmax和AUC0→t分别为3 h、1.19 h、1221.57 ng·mL-1和9983.40 ng·mL-1·h.结论:所建方法符合生物样品定量分析的基本要求,可用于水黄皮素在大鼠血浆中的含量测定及其药代动力学评价;水黄皮素在雌性SD大鼠体内的生物利用度更高.
Objective:To establish a quantitative analysis method for the determination of karanjin in rat plasma and study its pharmacokinetics.Methods:SD rats were intragastrically administered with karanjin(10 mg·kg-1).The plasma samples were pretreated by protein precipitation method and then detected quantitatively by HPLC-MS/MS.Schisandrin was employed as an internal standard.The separation was performed on an Agilent Poroshell 120 EC-C18(50 mm×3.0 mm,2.7 μm)column with acetonitrile(A)-0.1%methanoic acid solution(B)as mobile phase at a flow rate of 0.3 mL·min1.Electrospray ionization(ESI)source and positive ion multiple reaction monitoring(MRM)mode were used for scanning,and the results showed that karanjin[M+H]+m/z 293.1→277.0,schisandrin[M+H]+m/z 433.2→384.2;the pharmacokinetic parameters were calculated by DAS software.Results:In the range of 1.5625-1600 ng·mL-1,karanjin showed a good liner relationship in plasma(r=0.9997),with a low limit of quantification of 1.5625 ng·mL-1,precision(RSD)and accuracy(RE)were less than 20%.The precision RSD and accuracy RE of low medium and high quality control samples were less than 15%.The Tmax,t1/2,Cmax and AUC0→t of female SD rats were 3 h,1.95 h,1214.85 ng·mL-1 and 11920.43 ng·mL-1·h respectively,while that of male rats were 3 h,1.19 h,1221.57 ng·mL-1 and 9983.40 ng·mL-1·h respectively.Conclusion:The established method meets the basic requirements of quantitative analysis of biological samples,and can be used for the content determination and pharmacokinetic evaluation of karanjin in rat plasma.The bioavailability of karanjin in female SD rats is higher.