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Quantitative and Wide-Ranging Profiling of Phospholipids in Human Plasma by Two-dimensional Liquid Chromatography/Mass Spectrometry

机译:二维液相色谱/质谱法定量分析人血浆中的磷脂

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摘要

Normal-phase or reverse-phase liquid chromatographynhas been used in phospholipidomics for lipid separationnprior to mass spectrometry analysis. However, separationnusing a single separation mode is often inadequate, asnhigh-abundance phospholipids can mask large numbersnof low-abundance lipids of interest. In order to detect andnquantify low-abundance phospholipids, we present a novelntwo-dimensional (2D) approach for sensitive and quantitativenglobal analysis of phospholipids. The methodologynmonitors individual glycerolipids and phospholipidsnthrough the use of a new quantitative normal-phase, solidphasenextraction procedure, followed by molecular characterizationnand relative quantification using an ion-trapnOrbitrap equipped with a reverse-phase liquid chromatograph,nwith data processing by MS++ software. The CVn(%) of the peak area of each lipid standard was less thann15% with this extraction method. When the method wasnapplied to a liver sample, we could detect more phosphatidylserinen(PS) compared to the previous method.nFinally, our developed method was applied to Alzheimer’sndisease (AD) plasma samples. Several hundred peaksnwere detected from a 60 μL plasma sample. A partialleast-nsquares discriminant analysis (PLS-DA) plot usingnpeak area ratio gave a unique group of PLS scores whichncould distinguish plasma samples of Alzheimer’s diseasen(AD) patients from those of age-matched healthy controls.
机译:在质谱分析之前,正相或反相液相色谱已用于磷脂组学中的脂质分离。但是,使用单一分离模式进行分离通常是不够的,因为高丰度的磷脂可以掩盖大量的低丰度目的脂质。为了检测和定量低丰度的磷脂,我们提出了一种新颖的二维(2D)方法对磷脂进行灵敏和定量的全局分析。该方法通过使用新的定量正相固相萃取方法监测单个的甘油脂和磷脂,随后使用配备了反相液相色谱仪的离子阱Orbitrap进行分子表征和相对定量,并通过MS ++软件进行数据处理。用这种提取方法,每个脂质标准品的峰面积的CVn(%)小于n15%。当该方法应用于肝脏样品时,与以前的方法相比,我们可以检测到更多的磷脂酰丝氨酸(PS)。最后,我们开发的方法被应用于阿尔茨海默氏症(AD)血浆样品。从60μL血浆样品中检测到数百个峰。使用峰面积比的偏最小二乘平方判别分析(PLS-DA)图给出了一组独特的PLS评分,这无法区分阿尔茨海默氏病(AD)患者的血浆样品和年龄匹配的健康对照者。

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  • 来源
    《Analytical Chemistry》 |2010年第23期|p.9858-9864|共7页
  • 作者单位

    Eisai Co., Ltd., 5-1-3 Tokodai, Tsukuba, Ibaraki 300-2635, Japan, and Core Research for Evolutional Science andTechnology, Japan Science and Technology, Saitama 332-0012, Japan;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
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  • 入库时间 2022-08-17 13:36:54

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