固相萃取-液相色谱-串联质谱法测定人血浆中卢帕他定的浓度

Determination of Rupatadine in Human Plasma by Solid Phase Extraction and LC-MS/MS

目的:建立人血浆中卢帕他定浓度的液相色谱-串联质谱(LC/MS/MS)检测方法。方法:以氯雷他定为内标,血浆样本经0.1 mol/L氢氧化钠碱化后用固相小柱萃取处理;分析柱为Kromasil C18柱(150 mm×4.6 mm,5μm),保护柱为PhenomenexGemini C18柱(4 mm×3.0 mm,10μm),流动相为甲醇-10 mmol/L甲酸铵水溶液(85∶15);使用电喷雾离子源(ESI),正离子多反应监测(MRM)方式进行检测。结果:每个样本分析时间为5 min,血浆中内源性物质对测定无干扰,卢帕他定线性范围为0.011 9～15.000 0μg/L,回归方程Y=2.883X+0.009,r=0.999,定量下限为0.011 9μg/L。日内、日间精密度(RSD)均小于10%,卢帕他定与内标的平均回收率分别为102.0%和104.1%,平均基质效应为103.7%和106.2%,且均不存在浓度依赖性。结论:本方法特异性强,灵敏度高,测定结果可靠,适用于临床试验中血浆样本的高通量分析。

Objective： To establish LC-MS/MS method for the determination of rupatadine in human plasma by using a solid phase extraction technique. Methods： Loratadine was used as internal standard and plasma samples were alkalified by 0. 1 mol/L sodium hydroxide solution, and were extracted with solid phase extraction. The extractive was separated on a Kromasill C18 analytical column （150 mm×4.6 mm, 5μm） guarded by a Phemomenex Gemini C18 column （4 mm×3.0 mm, 10μm）. The mobile phase consisted of methanol-10 mmol/L ammonium formate （85 ： 15, v/v）. Electrospray ionization （ESI） source was applied and operated in the positive multiple reactions monitoring （ MRM ） mode. Results： Each analysis was completed within five minutes. Chromatogram showed no endogenous interfering peaks with blank samples. Good linearity was found within 0. 0119 - 15. 0000 p.g/L and with the regression equation of Y= 2. 883X＋0. 009, r = 0. 999. The limit of quantification was 0.0119 μg/L. The inter-and intra-day precision （RSD） were both less than 10%. The average recovery of rupatadine and loratadine were 102.0% and 104.1%, respectively. The average matrix effects of rupatadine and loratadine were 103.7% and 106.2% respectively, in which no concentration dependences were observed. Conclusions： The method is specific, sensitive and accurate, so it is suitable for the determination of rupatadine in human plasma of clinical samples.