Full automation of serotonin determination by column-switching and HPLC
Full automation of serotonin determination by column-switching and HPLC
复制标题
通过柱切换和 HPLC 全自动测定血清素
DOI：
10.1007/bf02270450
复制
发表时间：
1990
期刊：
影响因子：
1.7
通讯作者：
R. Pasquali
中科院分区：
文献类型：
--
作者：
G. Grossi;A. Bargossi;G. Sprovieri;V. Bernagozzi;R. Pasquali
本文介绍了一种简便、快速、全自动的血浆5-羟色胺测定方法。通过耦合两个设备实现全自动化：样品处理站和固相自动进样器。样品处理站稀释血浆样品，然后与固相自动进样器联机。它首先用样品净化所需的所有溶剂填充环路，然后反转流动，将这些溶剂泵送通过位于自动进样器上的二氧化硅键合阳离子交换一次性萃取柱。对于洗脱，将柱与HPLC分析柱一起切换运行，并通过HPLC流动相洗脱血清素。通过离子对反相液相色谱法进行HPLC分离。柱流出物通过电化学反应器完全还原，并且通过双电池电化学检测器以氧化模式检测5-羟色胺。血浆样品为50 μl，血浆灵敏度为40 ng/l，保留时间为6 min，回收率为95%。确定了贫血小板和富血小板血浆的重复性和正常范围，并计算了与手动HPLC的相关性。
SummaryA simple, fast, fully automated method for plasma serotonin determination is described. Full automation is obtained by coupling two devices: a sample processing station and a solid-phase autosampler. The sample processing station dilutes the plasma sample and is then connected, on-stream, with the solid-phase autosampler. It firstly fills a loop with all the solvents necessary for the sample clean-up, then, inverting the flow, pumps these solvents through the silica-bonded cation-exchange disposable extraction cartridge positioned on the autosampler. For the elution, the cartridge is switched on-stream with the HPLC analytical column and serotonin is eluted by the HPLC mobile-phase. The HPLC separation is performed by ion-pairing reversed-phase liquid chromatography. The column effluent is completely reduced by an electrochemical reactor and serotonin is detected in an oxidation-mode by a dual-cell electrochemical detector. The plasma sample is 50 μl, the plasma sensitivity is 40 ng/l, the retention time is 6 min and the recovery is 95%. The repeatibility, the normal ranges for platelet-poor and for platelet-rich plasma have been established and correlation with manual HPLC calculated.