Fast and Sensitive Detection of Oligosaccharides Using Desalting Paper Spray Mass Spectrometry (DPS-MS).
Fast and Sensitive Detection of Oligosaccharides Using Desalting Paper Spray Mass Spectrometry (DPS-MS).
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DOI：
10.1021/jasms.0c00310
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发表时间：
2020-10-07
影响因子：
3.2
通讯作者：
Chen H
中科院分区：
文献类型：
--
作者：
Wang Q;Bhattarai M;Zhao P;Alnsour T;Held M;Faik A;Chen H
传统的基于质谱仪(MS)的小碳水化合物片段(低聚糖，聚合度2-12)分析方法非常耗时，因为需要脱盐等离线样品前处理。在这里，我们报道了一种新的纸喷雾电离方法，称为除盐纸喷雾(DPS)，它使用一张三角滤纸来同时进行样品的除盐和电离。与常规的纸喷雾电离(PSI)和纳米电喷雾电离(NanESI)不同，DPS-MS允许快速、灵敏地检测具有复杂基质(例如Tris、PBS、HEPES缓冲液或尿液)的生物样品中的寡糖。当将低聚糖样品装载到以纤维素为主的滤纸基材(10 mm×5 mm，高度×底部)上时，低聚糖通过低聚糖与纤维素之间的亲水作用吸附在滤纸上。盐和缓冲液可以用ACN/H2O(90/10v/v)溶液洗掉，而寡糖可以用ACN/H2O/甲酸(FA)(10/90/1v/v/v)溶液从纸上洗脱出来，并直接从纸尖喷雾电离。用DPS-MS(每个样品5分钟)可以快速分析非挥发性缓冲液中微量(例如，50fmol)的各种糖类。DPS-MS还适用于直接检测糖基转移酶(GT)反应中的寡糖，这是一项具有挑战性的任务，通常需要进行放射性检测。受体和产物寡糖的定量分析表明，随着用于反应的GT酶的增加，产物增加，结果与放射性测定一致。这项工作表明，DPS-MS具有从生物样品中快速分析寡糖的潜力。
Conventional mass spectrometry (MS)-based analytical methods for small carbohydrate fragments (oligosaccharides, degree of polymerization 2–12) are time-consuming due to the need for offline sample pretreatment such as desalting. Herein, we report a new paper spray ionization method, named desalting paper spray (DPS), which employs a piece of triangular filter paper for both sample desalting and ionization. Unlike regular paper spray ionization (PSI) and nano electrospray ionization (nanoESI), DPS-MS allows fast and sensitive detection of oligosaccharides in biological samples having complex matrices (e.g., Tris, PBS, HEPES buffers or urine). When oligosaccharide sample is loaded onto the filter paper substrate (10 mm × 5 mm, height × base) made mostly of cellulose, oligosaccharides adsorb on the paper via hydrophilic interactions between oligosaccharides and cellulose. Salts and buffers can be washed away using an ACN/H2O (90/10 v/v) solution, While oligosaccharides can be eluted from the paper using a solution of ACN/H2O/formic acid (FA) (10/90/1 v/v/v) and directly spray-ionized from the tip of the paper. Various saccharides at trace levels (e.g., 50 fmol) in nonvolatile buffer can be quickly analyzed by DPS-MS (< 5 min per sample). DPS-MS is also applicable for direct detection of oligosaccharides from glycosyltransferase (GT) reactions, a challenging task which typically requires radioactive assay. Quantitative analysis of acceptor and product oligosaccharides shows increased product with increased GT enzymes used for the reaction, a result in line with radioactivity assay. This work suggests that DPS-MS has potential for rapid oligosaccharides analysis from biological samples.
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