Quantitation of carbohydrate monomers and dimers by liquid chromatography coupled with high-resolution mass spectrometry
Quantitation of carbohydrate monomers and dimers by liquid chromatography coupled with high-resolution mass spectrometry
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DOI：
10.1016/j.carres.2018.08.007
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发表时间：
2018-10-01
影响因子：
3.1
通讯作者：
Aristilde, Ludmilla
中科院分区：
文献类型：
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作者：
Barzen-Hanson, Krista A.;Wilkes, Rebecca A.;Aristilde, Ludmilla
作为植物废物或植物分泌物的残留物，碳水化合物广泛存在于各种环境基质中。含碳水化合物的原料是商品化合物工程生物生产的重要碳源。异质碳水化合物混合物的常规监测和定量需要快速、准确和精确的分析方法。在这里，我们提出了两种通过亲水相互作用液相色谱与电喷雾电离高分辨率质谱联用来量化碳水化合物混合物的方法。方法 1 针对 11 种不同的碳水化合物进行了优化：三种戊糖（核糖、阿拉伯糖、木糖）、三种己糖（葡萄糖、果糖、甘露糖）和五种二聚体（蔗糖、纤维二糖、麦芽糖、海藻糖、乳糖）。方法 1 可以同时监测这些碳水化合物，木糖/阿拉伯糖和乳糖/麦芽糖/纤维二糖峰共洗脱的情况除外。方法 2 使用与方法 1 相同的固定相和流动相来分离葡萄糖和半乳糖，而方法 1 中无法区分葡萄糖和半乳糖。这两种方法都具有较低的检测限 (0.019-0.40 μM) 和定量限 (0.090-1.3 μM)、良好的精密度 (2.4-13%)（蔗糖 (18%) 除外）和较低的质量误差 (0.0-2.4 ppm)。方法 1 在分析高离子强度溶液时效果良好，但观察到中等的基质效应。最后，我们应用方法 1 同时跟踪保护假单胞菌 Pf-5（一种生物技术上重要的土壤细菌物种）对五种碳水化合物（木糖、葡萄糖、果糖、甘露糖和麦芽糖）的细胞外消耗。
As remnants of plant wastes or plant secretions, carbohydrates are widely found in various environmental matrices. Carbohydrate-containing feedstocks represent important carbon sources for engineered bioproduction of commodity compounds. Routine monitoring and quantitation of heterogenous carbohydrate mixtures requires fast, accurate, and precise analytical methods. Here we present two methods to quantify carbohydrates mixtures by coupling hydrophilic interaction liquid chromatography with electrospray ionization high-resolution mass spectrometry. Method 1 was optimized for eleven different carbohydrates: three pentoses (ribose, arabinose, xylose), three hexoses (glucose, fructose, mannose), and five dimers (sucrose, cellobiose, maltose, trehalose, lactose). Method 1 can monitor these carbohydrates simultaneously, except in the case of co-elution of xylose/arabinose and lactose/maltose/cellobiose peaks. Using the same stationary and mobile phases as in Method 1, Method 2 was developed to separate glucose and galactose, which were indistinguishable in Method 1. Both methods have low limits of detection (0.019-0.40 mu M) and quantification (0.090-1.3 mu M), good precision (2.4-13%) except sucrose (18%), and low mass error (0.0-2.4 ppm). Method 1 was robust at analyzing high ionic strength solutions, but a moderate matrix effect was observed. Finally, we apply Method 1 to track concurrently the extracellular depletion of five carbohydrates (xylose, glucose, fructose, mannose, and maltose) by Pseudomonas protegens Pf-5, a biotechnologically-important soil bacterial species.