HPLC-MS/MS测定龙生蛭胶囊在大鼠血浆中7个活性成分的含量及其药代动力学研究

Determination of seven components of Longshengzhi capsules in rat plasma by HPLC-MS/MS and its application in pharmacokinetic study

目的:建立龙生蛭胶囊在血浆中活性成分的定量分析方法,并对各成分进行药代动力学研究。方法:采用HPLC-MS/MS分析方法,使用Agilent Rapid Resolution HD C_(18)色谱柱(50 mm×2.1 mm,1.8μm),流动相为乙腈-水(含0.1%甲酸),梯度洗脱,流速0.5 mL·min^(-1);采用电喷雾离子源,分别选择正或负离子模式进行多反应监检测扫描(MRM),分析时间正离子模式下6 min,负离子模式下3 min。结果:经方法学验证,各成分线性关系良好(r>0.9977),提取回收率在85.1%-114.5%范围内,基质效应在85.08%-112.95%范围内,日内精密度RSD<10%,日间精密度RSD<9.6%,稳定性RSD<13%,均符合生物样品分析要求,满足大鼠血浆样品中各成分分析测定的需求。用DAS 3.0软件计算龙生蛭胶囊的7个活性成分在大鼠体内的药代动力学参数。结果表明,苦杏仁苷、阿魏酸、异嗪皮啶、毛蕊异黄酮、洋川芎内酯A、氧化芍药苷、芍药苷的达峰时间(T_(max))分别为(0.36±0.31)h、(0.11±0.05)h、(0.10±0.04)h、(3.36±3.69)h、(1.07±0.80)h、(2.35±3.12)h、(0.60±0.44)h;消除半衰期(t_(1/2))分别为(1.02±0.46)h、(7.95±10.74)h、(32.55±15.57)h、(3.67±2.07)h、(1.19±0.12)h、(2.39±1.92)h、(1.50±0.12)h。结论:该研究为龙生蛭胶囊主要成分的药理活性研究及临床用药提供科学数据。

Objective:To establish a method for quantitative analysis of active components in plasma of Longshengzhi capsules and to study the pharmacokinetic of each component.Methods:Using HPLC-MS/MS analysis method,Agilent Rapid Resolution HD C_(18) column(2.1 mm×50 mm,1.8μm)was adopted and the mobile phase was acetonitrile-water(containing 0.1%formic acid)with gradient elution at a flow rate of 0.5 mL·min^(-1).The electrospray ion source was adopted,and the positive or negative ion mode for multiple reaction detection scan(MRM)was selected.In detection mode,the analysis time was 6 min in positive ion mode and 3 min in negative ion mode.Results:Methodologically verified,the linear relationship of each component was good(r>0.9977),the extraction recoveries were in the range of 85.1%-114.5%.The matrix effect was in the range of 85.08%-112.95%,the RSD values for intra-day precision,inter-day precision and stability were below 10%,9.6%and 13%,respectively.They all met the requirements of biological sample analysis and the requirements of analysis and determination of various components in rat plasma samples.The pharmacokinetic parameters of the 7 active ingredients of Longshengzhi capsules in rats were calculated by DAS 3.0 software.The results showed that the peak time(T_(max))of amygdalin,ferulic acid,isofraxidin,calycosin,senkyunolide A,oxypaeoniflora,and paeoniflorin was(0.36±0.31)h and(0.11±0.05)h,(0.10±0.04)h,(3.36±3.69)h,(1.07±0.80)h,(2.35±3.12)h and(0.60±0.44)h,respectively.The elimination half-life(t_(1/2))was(1.02±0.46)h,(7.95±10.74)h,(32.55±15.57)h,(3.67±2.07)h,(1.19±0.12)h,(2.39±1.92)h and(1.50±0.12)h,respectively.Conclusion:This study provides scientific data for the study of pharmacological activity and clinical use of the main components of Longshengzhi capsules.