两种前处理方法对LC - MS/MS测定家兔血清中淫羊藿黄酮类化合物基质效应的影响

摘要

An investigation was carried out for the impact of two pretreatment methods on the matrix effect of Epimedium-derived flavonoid compounds in rabbit serum by liquid chromatography - tandem mass spectrometry (LC - MS/MS). Before and after the pretreatment of rabbit blank serum via the extractions of ethyl acetate(liquid -liquid extraction, LLE)and C18 cartridge(solid-phase extraction, SPE ) respectively, a certain concentration solution of mixed reference substances containing four Epimedium-derived flavonoid compounds (icariin, icariside I , icariside H and iicaritin) was added exactly , and LC - MS/MS was adopted for sample analysis. Then, the absolute matrix effects, the recoveries and the process efficiencies of Epimedium-derived flavonoid compounds in rabbit serum were evaluated. The result showed that, after the sample pretreatment of LLE with ethyl acetate, the absolute matrix effects of Epimedium-derived flavonoid compounds in rabbit serum were more than 100% , the recoveries and the process efficiencies were much higher and stable, while after the sample pretreatment of SPE with C18 cartridge, the absolute matrix effects of Epimedium-derived flavonoid compounds in rabbit serum were less than 100% , the recoveries and the process efficiencies were much lower and unstable. The above results indicated that ionization enhancement can be realized for LC - MS/MS analysis of Epimedium-derived flavonoid compounds in rabbit serum while the recoveries and the process efficiencies of Epimedium-derived flavonoid compounds were much higher and more stable when using ethyl acetate for sample pretreatment, and this pretreatment method was also suitable for the pharmacokinetic study of Epimedium-derived flavonoid glycosides in rabbits as the serum sample pretreatment.%考察了两种前处理方法对采用液相色谱-串联质谱联用法( LC - MS/MS)测定家兔血清中淫羊藿黄酮类化合物时基质效应的影响.家兔空白血清分别以乙酸乙酯液-液萃取和C18小柱固相萃取,提取前、后各自加入一定浓度的4种淫羊藿黄酮类化合物的混合对照品溶液(淫羊藿苷、淫羊藿次苷I、淫羊藿次苷Ⅱ、淫羊藿素),评价家兔血清中淫羊藿黄酮类化合物的LC - MS/MS测定结果的绝对基质效应、提取回收率和方法过程效率.研究发现,以乙酸乙酯液-液萃取法进行样品前处理后,家兔血清中淫羊藿黄酮类化合物的绝对基质效应大于100％,提取回收率和方法过程效率均较高且稳定；以C18小柱固相萃取法进行样品前处理后,家兔血清中淫羊藿黄酮类化合物的绝对基质效应低于100％,提取回收率和方法过程效率均较低且不稳定.因此,采用乙酸乙酯液-液萃取法进行样品前处理,不仅可以增加LC - MS/MS测定家兔血清中淫羊藿黄酮类化合物的离子化效率,且对淫羊藿黄酮类化合物的提取回收率更高且稳定,适合作为样品前处理方法用于淫羊藿总黄酮苷在家兔体内的药代动力学研究.