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Revisiting monosaccharide analysis – quantitation of a comprehensive set of monosaccharides using dynamic multiple reaction monitoring†
Analyst ( IF 3.6 ) Pub Date : 2017-11-23 00:00:00 , DOI: 10.1039/c7an01530e
Gege Xu 1 , Matthew J Amicucci , Zhi Cheng , Ace G Galermo , Carlito B Lebrilla
Affiliation  

A rapid method for the quantitation of sixteen neutral and acidic monosaccharides, from both animal and plant sources was developed using ultra-high performance liquid chromatography triple quadrupole mass spectrometry (UHPLC/QqQ-MS) in dynamic multiple reaction monitoring (dMRM) mode. Monosaccharides including three pentoses (ribose, xylose, arabinose), two deoxyhexoses (rhamnose, fucose), five hexoses (fructose, mannose, allose, glucose, galactose), two hexuronic acids (glucuronic acid, galacturonic acid), and two N-acetyl-hexosamines (GlcNAc, GalNAc), were derivatized with 1-phenyl-3-methyl-5-pyrazolone (PMP), while underivatized sialic acids, Neu5Ac and Neu5Gc, were simultaneously analyzed with a 10-minute run. With the optimized UHPLC conditions, baseline separations of the isomers were achieved. The sensitivity and calibration ranges of this method were determined. The limits of detection were between femtomoles and attomoles with linear ranges spanning four to six orders of magnitude and coefficients of variation (CVs) ≤7.2%. Spiking experiments performed on a pooled fecal sample demonstrated the high accuracy of this method even when applied to samples with complicated matrices. The validated method was applied to fecal samples from an infant transitioning from breast milk to weaning foods. Major milk monosaccharides including galactose, fucose, glucose, GlcNAc, and Neu5Ac were found to be the most abundant components in the feces of milk-fed infants. PMP-derivatives of nine other monosaccharides including apiose, lyxose, altrose, talose, gulose, glucosamine, galactosamine, mannosamine, and N-acetylmannosamine (ManNAc) were also tested and could be added to the quantitation method depending on the need. The speed and sensitivity of the method makes it readily adaptable to rapid throughput analysis of monosaccharides in biological samples.

中文翻译:


重新审视单糖分析——使用动态多反应监测对一组全面的单糖进行定量†



使用超高效液相色谱三重四极杆质谱 (UHPLC/QqQ-MS) 在动态多反应监测 (dMRM) 模式下开发了一种快速定量来自动物和植物来源的 16 种中性和酸性单糖的方法。单糖包括三个戊糖(核糖、木糖、阿拉伯糖)、两个脱氧己糖(鼠李糖、岩藻糖)、五个己糖(果糖、甘露糖、阿洛糖、葡萄糖、半乳糖)、两个己糖醛酸(葡萄糖醛酸、半乳糖醛酸)和两个N-乙酰基-己糖胺(GlcNAc、GalNAc)用 1-苯基-3-甲基-5-吡唑啉酮 (PMP) 衍生化,而未衍生化的唾液酸、Neu5Ac 和 Neu5Gc 通过 10 分钟的运行同时进行分析。通过优化的 UHPLC 条件,实现了异构体的基线分离。确定了该方法的灵敏度和校准范围。检测限在飞摩尔和阿托摩尔之间,线性范围跨越四到六个数量级,变异系数 (CV) ≤ 7.2%。对汇集的粪便样本进行的加标实验证明,即使应用于具有复杂基质的样本,该方法也具有高精度。经过验证的方法适用于从母乳过渡到断奶食品的婴儿的粪便样本。研究发现,牛奶喂养的婴儿粪便中最丰富的成分是半乳糖、岩藻糖、葡萄糖、GlcNAc 和 Neu5Ac 等主要牛奶单糖。还测试了其他九种单糖的 PMP 衍生物,包括芹菜糖、来苏糖、阿卓糖、塔罗糖、古洛糖、葡萄糖胺、半乳糖胺、甘露糖胺和N-乙酰甘露糖胺 (ManNAc),并且可以根据需要添加到定量方法中。 该方法的速度和灵敏度使其很容易适用于生物样品中单糖的快速通量分析。
更新日期:2017-11-23
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