HPLC-MS/MS法测定鸡骨草与毛鸡骨草中主要成分在大鼠尿液和胆汁中的浓度及其排泄动力学比较研究

目的：建立同时测定大鼠尿液中9种成分和胆汁中6种成分的HPLC-MS/MS法，并对鸡骨草与毛鸡骨草中夏佛塔苷、异夏佛塔苷、相思子碱、芒柄花素、木犀草素、维采宁-2、下箴刺桐碱、牡荆素、异牡荆素在大鼠尿液和胆汁中的排泄动力学进行比较。方法：SD大鼠按一定剂量灌胃给予鸡骨草与毛鸡骨草提取物后，在24 h内收集胆汁，96 h内收集尿液。色谱分离采用ZORBAX SB-C₁₈（4.6 mm&#215;250 mm，5 μm）柱，流动相为甲醇-0.05%乙酸水溶液，梯度洗脱，流速1.0 mL&#183;min^-1；质谱检测采用电喷雾离子化（ESI）源，正负离子切换，多反应监测（MRM）模式。结果：尿液的胆汁中各待测成分在测定浓度范围内呈良好的线性关系（r≥0.996 6），日内、日间精密度RSD均小于15%，准确度RE在-11.2%~14.0%，提取回收率均不低于85.9%，基质效应基质因子RSD均不大于15%，样品稀释效应RSD<15%。实验结果表明，鸡骨草与毛鸡骨草中的多数成分在尿液中的排泄情况相似，但在胆汁中差异较大。结论：建立的HPLC-MS/MS方法简单、快速，灵敏度高。测定结果为鸡骨草与毛鸡骨草相互替代合理性的全面评价提供了一定的参考依据。

Objective: To establish an HPLC-MS/MS method for the simultaneous analysis of 9 ingredients in urine and 6 ingredients in bile after oral administration of Abri Herba and Abri Mollis Herba extract,and the comparative study of the compositions like hypaphorin,abrine,vicenin-2,schaftoside,isoschaftoside,vitexin,isovitexin,luteolin,formononetin,and sulfamethoxazole excretion kinetics.Methods: After a single gavage administration of Abri Herba and Abri Mollis Herba extract given to rats,bile samples were collected within 24 h and urine samples were collected within 96 h.The chromatographic separation was performed using ZORBAX SB-C₁₈ column(4.6 mm&#215;250 mm,5 μm),which was eluted with a gradient of methanol and 0.05% acetic acid (v/v) at a flow rate of 1.0 mL&#183;min^-1. The detection of the target compounds was performed in multiple-reaction monitoring (MRM) mode using a hybrid triple quadrupole linear ion trap mass spectrometer equipped with positive/negative ion-switching ESI source.Results: The calibration curves exhibited good linearity for all the compounds within the concentration range with correlation coefficients(r)≥0.996 6.The intra-day and inter-day precisions (RSD) were both less than 15.0% and the accuracies (RE) ranged from -11.2% to 14.0%.The mean extraction recoveries were both no less than 85.9% and the matrix factor of the matrix effects (RSD) were not more than 15%.Dilution effect (RSD) was less than 15%.All of the validation met the requirements for biopharmaceutical analysis.Conclusion: The developed method was simple,selective and specific.The results of this study provided a certain theoretical basis for the comprehensive evaluation of the rationality of the substitution between Abri Herba and Abri Mollis Herba.