LC-MS/MS同时测定犬干血浆样品中栀子苷、山栀苷及京尼平龙胆双糖苷的浓度

目的：建立LC-MS/MS法同时测定犬干血浆样品中栀子苷、山栀苷及京尼平龙胆双糖苷浓度的分析方法，并应用于犬药代动力学研究中。方法：采用Ecosil C18 （4.6 mm×150 mm，5 μm）色谱柱，流动相为甲醇-20 mmol/L 甲酸铵（0.1%甲酸），梯度洗脱；使用电喷雾离子源（ESI），正离子多反应监测模式检测。应用干血浆采样技术收集犬静脉输注栀子提取液后的血浆样品，超声提取后进行LC-MS/MS分析。结果： 3种环烯醚萜苷类成分在线性范围内的相关性良好（r〉0.99），日内、日间变异均〈15%，绝对回收率在 79.9%~91.4%之间，4 ℃ 放置和室温放置 24 h、75 d 稳定性良好。药代动力学参数显示栀子苷、山栀苷、京尼平龙胆双糖苷的t1/2分别为 6.86、0.687、2.11 h，AUC0→t分别为158、9.16、34.0 μg·mL^-1·h。结论：建立的同时检测干血浆样品中栀子苷、山栀苷、京尼平龙胆双糖苷浓度的液相色谱-串联质谱分析法，灵敏度好、专属性强。干血浆样品室温放置稳定性好，易于储存运输，适用于药代动力学研究中。

Simultaneous determination of geniposide,shanzhiside and genipin geniobioside concentration in dog dried plasma spots by LC-MS/MS

AIM：To establish a method that combine dried plasma spots and LC-MS/MS for simultaneous determination geniposide,shanzhiside and genipin geniobioside in dog plasma and study pharmacokinetic in dogs. METHODS：Chromatographic separation was achieved on an Ecosil C18（4.6mm×150mm,5μm）column and the mobile phase consisting of methanol and 20mmol/L ammonium formate（containing 0.1%formic acid）,gradient elution.An electrospray ionization（ESI）source was applied and operated in the positive multiple reaction monitoring（MRM）mode.Using dried plasma spots technology collect plasma samples after intravenous administration of Zhizi.The assay employed simple solvent extraction and sonication of the DPS sample,followed by LC-MS/MS system for analysis.RESULTS：Good linearity was achieved for the three compounds（r〉0.99）,the intraday and inter-day precision（RSDs）were below 15%,the absolute recoveries were between 79.9%-91.4%.The main pharmacokinetic parameters of geniposide,shanzhi side,genipin geniobioside as follows：the t1/2were 6.86,0.687,2.11 h,the AUC0→t were 158,9.16,34.0μg·mL^-1·h. CONCLUSION：The developed LC-MS/MS method is sensitive and specific for simultaneous determination of geniposide,shanzhiside and genipin geniobioside in dried plasma spots.The dried plasma spots were stable at room temperature,easy transportation and preservation,and it can be used for pharmacokinetic research.