摘要
该研究建立了一种简便、快速、灵敏的超高效液相色谱-串联质谱法(UPLC-MS/MS),能够同时测定给药炒酸枣仁提取物后比格犬血浆中木兰花碱、乌药碱、维采宁Ⅱ、异斯皮诺素、斯皮诺素、当药黄素、N-去甲基荷叶碱、6‴-阿魏酰斯皮诺素和酸枣仁皂苷B的含量.采用Waters HSS-T3 C18 色谱柱(2.1 mm×100 mm,1.8 μm),流动相为甲醇-水(含0.01%甲酸),梯度洗脱,9 种成分和 2 种内标于 8 min内完全分离;质谱检测在电喷雾离子源正、负离子切换模式下,采用多反应监测模式进行检测.分析方法经特异性、选择性、线性范围、准确度、精密度、回收率、基质效应和稳定性验证,可满足犬灌胃炒酸枣仁提取物后的药代动力学的研究要求.结果显示,比格犬单次灌胃给药后,木兰花碱、乌药碱、维采宁Ⅱ、异斯皮诺素、斯皮诺素、6‴-阿魏酰斯皮诺素和酸枣仁皂苷B的Tmax 为2.40～3.20 h,t1/2 为2.08～6.79 h,其中木兰花碱和斯皮诺素的暴露量相对较高,Cmax 分别为 76.7、31.5 ng·mL-1,AUC0-∞分别为 581、315 ng·h·mL-1;其余 5 种暴露较弱的化合物的 Cmax 为 0.81～13.0 ng·mL-1,AUC0-∞为 6.00～106 ng·h·mL-1.该研究为炒酸枣仁后续相关药理研究提供了参考依据.
Abstract
This study established a convenient,rapid,and sensitive ultra-performance liquid chromatography tandem mass spec-trometry(UPLC-MS/MS)method for simultaneous determination of magnoflorine,(R)-coclaurine,vicenin Ⅱ,isospinosin,spinosin,swertisin,N-nornuciferine,6‴-feruloylspinosin,and jujuboside B in beagle dog plasma after oral administration of fried Ziziphi Spi-nosae Semen(FZSS)extract.The Waters HSS-T3 C18 column(2.1 mm×100 mm,1.8 μm)was used.The methanol-aqueous solu-tion(containing 0.01%formic acid)was adopted as the mobile phase for gradient elution.The nine components and two internal standards were completely separated within 8 min.The mass spectrometry detection was performed in multiple reaction monitoring(MRM)mode by positive and negative ion switching of electrospray ionization.The analytical method was validated in terms of speci-ficity,selectivity,linear range,accuracy,precision,recovery,matrix effect,and stability.It could meet the requirement of pharmaco-kinetic research after oral administration of FZSS extract to beagle dogs.The results showed that the time to reach the peak concentra-tion(Tmax)of magnoflorine,(R)-coclaurine,vicenin Ⅱ,isospinosin,spinosin,6‴-feruloylspinosin,and jujuboside B was 2.40-3.20 h,and the elimination halflife(t1/2)was 2.08-6.79 h after a single-dose oral administration of FZSS to beagle dogs.The exposure of magnoflorine and spinosin was high,with a peak concentration(Cmax)of 76.7 and 31.5 ng·mL-1 and an area under the curve(AUC0-∞)of 581 and 315 ng·h·mL-1,respectively.The exposure of the remaining five compounds was lower,with a Cmax of 0.81-13.0 ng·mL-1 and an AUC0-∞ of 6.00-106 ng·h·mL-1.This study provides a reference for the follow-up research of FZSS.
基金项目
中国中医科学院科技创新工程项目(CI2021A04910)
中国中医科学院科技创新工程项目(CI2021B015)
NETL
NSTL
万方数据