LC-MS/MS法测定人血浆中白藜芦醇的浓度

目的:建立液相色谱-串联质谱法测定人血浆中白藜芦醇浓度的方法。方法:血浆样品经四硼酸钠溶液碱化后,经甲基叔丁基醚提取处理。分析柱为RestekC1(82.1mm×150mm,5.0μm),保护柱为GminiC1(84mm×3.0mm,10.0μm),流动相为乙腈-水(85∶15,V/V,其中含0.2mmol·L-1乙酸铵),流速为0.3mL·min-1,进样量为5μL,内标为3,5-二羟基-4-异丙基二苯乙烯,使用电喷雾离子源,以负离子多重反应监测(MRM)方式进行检测。用于定量白藜芦醇和内标的MRM扫描离子通道分别为m/z227.0→143.0、253.2→211.0。每个样品的分析时间为5min。结果:白藜芦醇和内标的保留时间分别为1.43、2.55min。血浆中内源性物质对测定无干扰,白藜芦醇检测浓度在0.1～50.0ng·mL-1范围内同其与峰面积之比呈良好线性关系(r=0.9969),定量下限为0.1ng·mL-1。日内、日间RSD均低于11.25%,方法回收率为(91.70±3.81)%～(99.00±3.29)%。结论:本方法特异性强,灵敏度高,测定结果可靠,适用于白藜芦醇的血药浓度测定。

Determination of the Concentration of Resveratrol in Human Plasma by LC-MS/MS

OBJECTIVE： To establish a LC-MS/MS method for the determination of resveratrol concentration in human plasma. METHODS： After alkalified by sodium tetraborate solution, the plasma samples were extracted with methyl tert-butyl ether. The sample was separated on Restek C18 column （2.1 mm×150 mm,5.0 μm） guarded by a Gmini C18 column（4 mm×3.0 mm,10.0 μm）. The mobile phase consisted of acetonitrile-water （85 ∶ 15,V/V,containing 0.2 mmol·L-1 ammonium acetate） at the flow rate of 0.3 mL·min-1. The injection volume was 5 μL and 3,5-dihydroxy-4-isopropyl toluylene was used as the internal standard （IS）. Electrospray ionization （ESI） source was applied and operated in the negative multiple reaction monitoring （MRM） mode. Resveratrol and IR were detected on MRM by the transitions from the precursor to the product ion （m/z 227.0→143.0、253.2→211.0）. And the total running time was 5 min. RESULTS： The retention time of resveratrol and IS were 1.43 min and 2.55 min. Chromatograms showed no endogenous interfering peaks with blank samples. The linear range of resveratrol was 0.1～50.0 ng·mL-1（r=0.996 9）. The limit of quantification was 0.1 ng·mL-1. The RSDs of inter-day and intra-day were less than 11.25% and the average recoveries were （91.70±3.81）%～（99.00±3.29）%. CONCLUSIONS： The method is specific, sensitive and accurate, and it is suitable for plasma concentration determination of resveratrol.