本研究建立了一个方便,快速,和灵敏的超高效液相色谱-串联质谱(UPLC-MS/MS)方法同时测定山药,(R)-牛磺酸,维可宁Ⅱ号,异spinosin,spinosin,swertisin,N-去甲亮氨酸,6-阿魏酸基多糖素,口服油炸酸枣仁(FZSS)提取物后,比格犬血浆中的枣甙B。WatersHSS-T3C_(18)柱(2.1mm×100mm,使用1.8μm)。采用甲醇-水溶液(含0.01%甲酸)作为梯度洗脱的流动相。九种组分和两种内标在8分钟内完全分离。通过电喷雾电离的正离子和负离子切换在多反应监测(MRM)模式下进行质谱检测。分析方法在特异性方面进行了验证,选择性,线性范围,准确度,精度,recovery,基体效应,和稳定性。Bagle犬口服FZSS提取物后,可以满足药代动力学研究的要求。结果表明,在达到最大浓度(T_(max))时,(R)-牛磺酸,维可宁Ⅱ号,异spinosin,spinosin,6-阿魏酸基多糖素,大枣苷B为2.40-3.20h,对比格犬单次口服FZSS后,消除半衰期(t_(1/2))为2.08-6.79h。马格诺氟林和spinosin的暴露量很高,峰值浓度(C_(max))为76.7和31.5ng·mL〜(-1),曲线下面积(AUC_(0-∞))为581和315ng·h·mL〜(-1),分别。其余5种化合物的暴露量较低,C_(max)为0.81~13.0ng·mL~(-1),AUC_(0-∞)为6.00~106ng·h·mL~(-1)。本研究为FZSS的后续研究提供了参考。
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.