LC-MS/MS法同时测定大鼠血浆中6种独一味成分

Simultaneous determination of 6 components from Lamiophlomis rotata in rat plasma by LC-MS/MS

目的:开发建立独一味药材中芹菜素、木犀草素、木犀草苷、绿原酸、毛蕊花糖苷和连翘酯苷B6种成分在大鼠血浆中的同步定量分析方法。方法:采用蛋白沉淀法进行样品前处理,选用安捷伦ZORBAX SB-C18(2.1mm×150mm i.d.,3.5μm)色谱柱及梯度洗脱法进行色谱分离,运用液相色谱串联质谱(LC-MS/MS)技术在多重反应监测(MRM)模式下进行样品测定,并从专属性、线性范围、准确度、精密度、回收率、介质效应和稳定性等方面对方法学进行了系统的验证。结果:在13min的分析时间内,能够实现对6种目标化合物的准确定量,方法学验证中的各项指标均符合生物样品分析的要求。结论:所建立的分析方法准确、灵敏、可靠,可用于研究独一味药材给药后相应成分在大鼠血浆中的暴露情况,为药材的质量分析和控制提供科学依据,也为该药材体内过程和效应机制的研究奠定了基础。

Objective： To develop and establish a quantitation method for the simultaneous determination of apigenin, luteolin, cynaroside, chlorogenic acid, verbascoside and forsythoside B from Lamiophlomis rotata in rat plasma. Methods： Simple protein precipitation was used for sample pre-treatment, an Agilent ZORBAX SB-C18 （2.1mmxl50mm i.d., 3.5μm） column and a gradient elution method were chosen for chromatographic separation, tandem mass spectrometer （LC-MS/MS） technique using multiple reaction monitoring mode （MRM） was applied for determination; a systematic method validation was performed through specificity, linear range, accuracy, precision, recovery, matrix effect and stability. Results： Accurate quantitation of the 6 components was realized within an analytical run time of 13min, and in method validation, each index met the required criteria of bio-sample analysis. Conclusion： The analytical method established was accurate, precise and reliable, which could be used to reveal the plasma exposure after rat administration of Lamiophlomis rotata, provide further scientific evidence to its quality analysis and control, and also promote the physiological disposition and pharmacodynamic mechanism study of this medicinal material.