大鼠灌胃当药苦苷后血浆中含氮代谢物R⁃gentiandiol和S⁃gentiandiol药代动力学研究

Pharmacokinetics of nitrogen⁃containing metabolites R⁃gentiandiol and S⁃gentiandiol in rat plasma after oral administration of swertiamarin

目的:建立血浆中当药苦苷的一对对映异构含氮代谢物R‑gentiandiol和S‑gentiandiol的手性拆分方法,并对这两个异构体的药代动力学进行研究。方法:采用LC‑MS/MS对当药苦苷的体内代谢物进行检测,使用Astec CyclobondⅡ环糊精手性色谱柱(4.6 mm×100 mm,5μm),选择乙腈‑水为流动相的梯度洗脱方式,且在正离子模式下采用多级反应监测展开分析。研究采用的定量分析反应离子对分别为R‑gentiandiol(m/z 210.04→192.06)、S‑gentiandiol(m/z 210.04→192.06),内标龙胆碱酮(m/z 192.02→162.08)。结果:两种含氮代谢物的大鼠血浆样品测定方法的线性相关系数均大于0.999,精密度小于7.00%,回收率为99.57%~102.65%,基质效应为90.94%~91.34%。当药苦苷灌胃大鼠后血浆中R‑gentiandiol和S‑gentiandiol达峰时间tmax分别为(1.63±0.23)、(1.58±0.21)h;半衰期t1/2分别为(6.23±0.52)、(5.46±0.38)h,最大血药浓度Cmax分别为(86.79±20.81)、(60.72±18.95)ng/mL,药时曲线下面积AUC0‑24分别为(1094.58±86.37)、(724.67±58.38)(ng·h)/mL。结论:该方法灵敏度高、精确度好,成功用于当药苦苷的体内代谢物R‑gentiandiol和S‑gentiandiol的手性拆分及其药代动力学研究,本研究方法及实验结果可为当药苦苷药效作用实体研究和药效物质基础研究提供科学依据,为中药单体的临床应用及资源开发奠定基础。

Objective:A chiral resolution method for enantiomers of two chiral nitrogen‑containing metabolites R‑gentiandiol and S‑gentiandiol of swertiamarin in plasma was developed,and the pharmacokinetics of the metabolites were studied.Meth⁃ods:The metabolites of swertiamarin in vivo were detected by LC‑MS/MS using Astec CyclobondⅡCyclodextrin column(4.6 mm×100 mm,5μm),gradient elution with acetonitrile‑water as mobile phase,and monitored by multiple reaction monitoring(MRM)method in positive mode.The ion pairs for quantitative analysis are R‑gentiandiol(m/z 210.04→192.06),S‑gentiandiol(m/z 210.04→192.06)and gentianone(m/z 192.02→162.08).Results:The linear correlation coefficients of the method devel‑oped were greater than 0.999,the precision was less than 7.00%,the recovery was 99.57%‑102.65%,and the matrix effect was 90.94%‑91.34%.The peak tmax of R‑gentiandiol and S‑gentiandiol in rat plasma after oral administration of swertiamarin were(1.63±0.23 h and(1.58±0.21)h,t1/2 was(6.23±0.52)h and(5.46±0.38)h,Cmax was(86.79±20.81)ng/mL and(60.72±18.95)ng/mL,and the AUC0‑24 were(1094.58±86.37))(ng·h)/mL and(724.67±58.38)(ng·h)/mL,respectively.Conclu⁃sion:The method was highly sensitive with good accuracy and precision,and it was successfully applied for chiral resolution and pharmacokinetics study of R‑gentiandiol and S‑gentiandiol in plasma.The method developed and experimental results will provide scientific basis for the study of pharmacodynamics and pharmacodynamic material basis of swertiamarin,and lay a foundation for clinical application and resource development of TCM monomer.