基于UPLC-MS/MS测定比格犬血浆中炒酸枣仁提取物9种成分及其药代动力学研究

Determination of nine components of fried Ziziphi Spinosae Semen extract in beagle dog plasma by UPLC-MS/MS and its application in pharmacokinetic research

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 C_(18)色谱柱(2.1 mm×100 mm,1.8μm),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T_(max)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,C_(max)分别为76.7、31.5 ng·mL^(-1),AUC_(0-∞)分别为581、315 ng·h·mL^(-1);其余5种暴露较弱的化合物的C_(max)为0.81~13.0 ng·mL^(-1),AUC_(0-∞)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。

This study established a convenient, rapid, and sensitive ultra-performance liquid chromatography tandem mass spectrometry(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 Spinosae Semen(FZSS) extract. The Waters HSS-T3 C_(18) column(2.1 mm×100 mm, 1.8 μm) was used. The methanol-aqueous solution(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 specificity, selectivity, linear range, accuracy, precision, recovery, matrix effect, and stability. It could meet the requirement of pharmacokinetic research after oral administration of FZSS extract to beagle dogs. The results showed that the time to reach the peak concentration(T_(max)) of magnoflorine,(R)-coclaurine, vicenin Ⅱ, isospinosin, spinosin, 6'''-feruloylspinosin, and jujuboside B was 2.40-3.20 h, and the elimination halflife(t_(1/2)) 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(C_(max)) of 76.7 and 31.5 ng·mL^(-1) and an area under the curve(AUC_(0-∞)) of 581 and 315 ng·h·mL^(-1), respectively. The exposure of the remaining five compounds was lower, with a C_(max) of 0.81-13.0 ng·mL^(-1) and an AUC_(0-∞) of 6.00-106 ng·h·mL^(-1). This study provides a reference for the follow-up research of FZSS.