柱前衍生化HPLC-MS/MS法测定人血浆中5-HT、AD和NA方法的建立

Determination of 5-hydroxytryptamine, Adrenalin and Noradrenalin in Human Plasma by Pre-column Derivatization HPLC-MS/MS

中文关键词:  5-羟色胺肾上腺素去甲肾上腺素柱前衍生化高效液相色谱-串联质谱法血药浓度

英文关键词:5-hydroxy tryptamineAdrenalinNoradrenalinPre-column derivatization HPLC-MS/MSPlasma concentration

中文摘要:

      目的建立衍生化高效液相色谱-串联质谱法测定人血浆中5-羟色胺（5-HT）、肾上腺素（AD）和去甲肾上腺素（NA）质量浓度。方法采用Ultimate C18 (50 mm×4.6 mm, 5 μm)色谱柱，流动相为乙腈∶水∶甲酸为95∶5∶0.05（V∶V∶V），流速为0.2 mL/min，柱温为室温。检测方式为多反应离子监测，正离子模式，电喷雾离子源。血浆样品经沉淀蛋白后，在pH 10.5条件下与丹酰氯发生衍生化反应，产物经乙酸乙酯萃取浓集后进样。结果标准曲线线性范围为2.5~200 ng/mL；日内精密度相对标准偏差（RSD）为1.33%~8.13%，日间RSD 2.86%~7.73%，均小于9%；方法回收率95.44%~109.71%，基质效应4.86%~12.81%。结论本法简单、快速，可用于人血浆中5-HT、AD和NA质量浓度测定和研究。

英文摘要:

      Objective to establish a method for the determination of 5-hydroxy tryptamine（5-HT), adrenalin （AD) and noradrenalin （NA) in human plasma using pre-column derivatization HPLC-MS/MS. Methods Derivatives of the plasma samples were produced with dansyl chloride under pH 10.5, and then extracted and enriched with ethyl acetate. The detected chemicals were separated using Ultimate C18 (50 mm×4.6 mm, 5 μm) with acetonitrile-water-formic acid=95∶5∶0.05 （V∶V∶V） at 0.2 mL/min. The HPLC-MS//MS system was operated under a multiple reaction monitoring mode (MRM) using the electrospray ionization technique in positive mode. ResultsLinear range of the calibration curve appeared in 250-2.5 ng/mL. The method had a less than 9% intra- and inter-assay relative standard deviation (RSD), 95.44%-109.71% recoveries, and 4.86%-12.81% matrix effects. ConclusionThis method is simple, accurate, and suitable for detection of 5-HT, AD and NA in human plasma.