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基于超临界流体萃取-超临界流体色谱/质谱的高通量磷脂分析系统用于干血浆斑点分析。

High-throughput phospholipid profiling system based on supercritical fluid extraction-supercritical fluid chromatography/mass spectrometry for dried plasma spot analysis.

机构信息

Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.

出版信息

J Chromatogr A. 2012 Aug 10;1250:69-75. doi: 10.1016/j.chroma.2012.06.031. Epub 2012 Jun 17.

Abstract

The dried blood spots (DBS) and dried plasma spots (DPS) analysis is available as a diagnostic tool for genetic diseases and is useful for the screening of biomarkers. In this study, a high-throughput analytical system based on supercritical fluid extraction-supercritical fluid chromatography with tandem mass spectrometry (SFE-SFC/MS/MS) was constructed for phospholipids profiling of DPS. This system is able to simultaneously perform extraction and separation, allowing phospholipids that have common polar head groups to be analyzed. Phospholipids in only 3 μl of plasma can be extracted in 5 min and analyzed within 15 min using this system. A total of 134 phospholipids, including phosphatidylcholine, lysophosphatidylcholine, sphingomyelin, phosphatidylethanolamine and lysophosphatidylethanolamine, were annotated, and 74 phospholipids were analyzed with good repeatability. The SFE-SFC/MS/MS, which is able to perform high-throughput lipid profiling analysis for clinical diagnosis and drug discovery, may also be suitable for the screening of biomarkers.

摘要

干血斑(DBS)和干血浆斑(DPS)分析可用作遗传疾病的诊断工具,并且对生物标志物的筛选很有用。在这项研究中,构建了一种基于超临界流体萃取-超临界流体色谱-串联质谱(SFE-SFC/MS/MS)的高通量分析系统,用于 DPS 中磷脂的分析。该系统能够同时进行提取和分离,使具有共同极性头基团的磷脂能够被分析。使用该系统,仅 3 μl 的血浆中的磷脂可以在 5 分钟内提取,并在 15 分钟内分析。总共注释了 134 种磷脂,包括磷脂酰胆碱、溶血磷脂酰胆碱、神经鞘磷脂、磷脂酰乙醇胺和溶血磷脂酰乙醇胺,并对 74 种磷脂进行了良好的重复性分析。该 SFE-SFC/MS/MS 能够对临床诊断和药物发现进行高通量脂质分析,也可能适用于生物标志物的筛选。

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