Mass spectrum (MS) is one of the most commonly used tools for qualitative and quantitative analysis of glycans. However, due to the complexity of biological samples and the low ionization efficiency of glycans, these need to be purified and derivatized prior to MS analysis. Existing purification strategies require a combination of multiple methods and are cumbersome to operate. Here, we propose a new method for the purification of glycoprotein N/O-glycans and their derivatives using a hand-packed absorbent cotton hydrophilic interaction chromatography column (HILIC). The method's reliability and applicability were verified by purifying N/O-glycans and the derivatives of standard glycoproteins, such as chicken albumin and porcine stomach mucin. Stable isotope labelling was used to compare the glycans' recovery following different purification methods. Absorbent cotton HILIC was also successfully applied for the analysis of human serum and fetal bovine serum glycoprotein N-glycans. Finally, testing revealed high binding capacity (9 mg/g-1 maltohexaose/absorbent cotton) and good recovery (average recovery was 91.7%) of glycans. Compared with traditional procedures, the proposed purification method offers considerable advantages, such as simplicity, high efficiency, economy, universality, and broad applicability for the pretreatment of glycans and their derivatives in biological samples prior to MS analysis.

中文翻译：

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通过吸收棉亲水色谱柱从生物样品中纯化N-和O-聚糖及其衍生物。

质谱（MS）是对聚糖进行定性和定量分析的最常用工具之一。但是，由于生物样品的复杂性和聚糖的低电离效率，需要在MS分析之前对它们进行纯化和衍生化。现有的纯化策略需要多种方法的组合，并且操作繁琐。在这里，我们提出了一种使用手工填充的吸收棉亲水相互作用色谱柱（HILIC）纯化糖蛋白N / O-聚糖及其衍生物的新方法。通过纯化N / O-糖和鸡白蛋白和猪胃粘蛋白等标准糖蛋白衍生物，验证了该方法的可靠性和适用性。稳定的同位素标记用于比较聚糖的 按照不同的纯化方法回收。吸收棉HILIC还成功地用于分析人血清和胎牛血清糖蛋白N-聚糖。最后，测试显示了高结合能力（9 mg / g-1麦芽六糖/吸收棉）和良好的聚糖回收率（平均回收率为91.7％）。与传统方法相比，所提出的纯化方法具有相当大的优势，例如简单，高效，经济，通用性强，在MS分析之前用于生物样品中聚糖及其衍生物的预处理具有广泛的适用性。测试显示了高结合能力（9 mg / g-1麦芽六糖/吸收性棉）和良好的聚糖回收率（平均回收率为91.7％）。与传统方法相比，所提出的纯化方法具有相当大的优势，例如简单，高效，经济，通用性强，在MS分析之前用于生物样品中聚糖及其衍生物的预处理具有广泛的适用性。测试显示了高结合能力（9 mg / g-1麦芽六糖/吸收性棉）和良好的聚糖回收率（平均回收率为91.7％）。与传统方法相比，所提出的纯化方法具有相当大的优势，例如简单，高效，经济，通用性强，在MS分析之前用于生物样品中聚糖及其衍生物的预处理具有广泛的适用性。