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通过液相色谱-串联质谱法对可溶性糖和糖醇进行定量分析。

Quantification of Soluble Sugars and Sugar Alcohols by LC-MS/MS.

作者信息

Feil Regina, Lunn John Edward

机构信息

Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany.

出版信息

Methods Mol Biol. 2018;1778:87-100. doi: 10.1007/978-1-4939-7819-9_7.

Abstract

Sugars are simple carbohydrates composed primarily of carbon, hydrogen, and oxygen. They play a central role in metabolism as sources of energy and as building blocks for synthesis of structural and nonstructural polymers. Many different techniques have been used to measure sugars, including refractometry, colorimetric and enzymatic assays, gas chromatography, high-performance liquid chromatography, and nuclear magnetic resonance spectroscopy. In this chapter we describe a method that combines an initial separation of sugars by high-performance anion-exchange chromatography (HPAEC) with detection and quantification by tandem mass spectrometry (MS/MS). This combination of techniques provides exquisite specificity, allowing measurement of a diverse range of high- and low-abundance sugars in biological samples. This method can also be used for isotopomer analysis in stable-isotope labeling experiments to measure metabolic fluxes.

摘要

糖类是主要由碳、氢和氧组成的简单碳水化合物。它们在新陈代谢中作为能量来源以及用于合成结构和非结构聚合物的构建单元发挥着核心作用。许多不同的技术已被用于测量糖类,包括折射法、比色法和酶法分析、气相色谱法、高效液相色谱法以及核磁共振光谱法。在本章中,我们描述了一种方法,该方法将通过高效阴离子交换色谱法(HPAEC)对糖类进行初步分离与串联质谱法(MS/MS)检测和定量相结合。这种技术组合提供了极高的特异性,能够测量生物样品中各种高丰度和低丰度的糖类。该方法还可用于稳定同位素标记实验中的同位素异构体分析,以测量代谢通量。

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