MONOSACCHARIDE ANALYSIS OF GLYCOCONJUGATES BY ANION-EXCHANGE CHROMATOGRAPHY WITH PULSED AMPEROMETRIC DETECTION
MONOSACCHARIDE ANALYSIS OF GLYCOCONJUGATES BY ANION-EXCHANGE CHROMATOGRAPHY WITH PULSED AMPEROMETRIC DETECTION
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DOI：
10.1016/0003-2697(88)90089-9
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发表时间：
1988-04-01
影响因子：
2.9
通讯作者：
LEE, YC
中科院分区：
文献类型：
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作者：
HARDY, MR;TOWNSEND, RR;LEE, YC
阴离子交换层析和脉冲安培检测的方法(AE-PAD；Johnson，D.C.和Polta，T.Z.(1986)Chromatogr.论坛1，37-44)已被应用于糖共轭化合物的组成分析。以22 mM氢氧化钠为柱流出物，未衍生岩藻糖、氨基半乳糖、氨基葡萄糖、半乳糖和甘露糖可在15min内以1ml/min的流速快速分离。单糖的定量下限优于100pmol(信噪比184：1)。用2M三氟乙酸水解液对几种糖蛋白、糖肽和低聚糖的单糖含量进行了测定。使用AE-PAD，中性糖和氨基糖可以在单一色层步骤中快速测定。完全释放N-乙酰氨基葡萄糖需要更有力的水解条件(Lee，Y.C.(1972)in Methods in Enzymology(Ginsburg，V.，Ed.)，Vol.28，pp.63-73，Academy Press，New York)。在糖肽和寡糖中，检测到的人的残留量比预测的少了大约一个。具有荧光检测的单糖复合体(Mikami，H.和Ishida，Y.(1983)Bunseki Kagaku 32，E207-E210)的AE-PAD和液相色谱分析都给出了类似的甘露糖和其他糖的定量。快速、灵敏地定量未衍生化的单糖的能力应有助于糖共轭化合物的结构分析。
The method of anion exchange chromatography followed by pulsed amperometric detection (AE-PAD; Johnson, D.C., and Polta, T.Z. (1986) Chromatogr. Forum 1, 37-44) has been applied to the compositional analysis of glycoconjugates. Using 22 mM naOH as a column effluent, underivatized fucose, galactosamine, glucosamine, galactose, and mannose were readily separated in 15 min at a flow rate of 1 ml/min. The limit of quantification of the monosaccharides was better than 100 pmol (signal to noise ratio 184:1). AE-PAD was employed to quantify the monosaccharides of several glycoproteins, glycopeptides, and oligosaccharides after hydrolysis with 2 M trifluoroacetic acid. Both neutral and amino sugars could be rapidly estimated in a single chromatographic step using AE-PAD. Complete release of N-acetylglucosamine required more vigorous hydrolysis conditions (Lee, Y. C. (1972) in Methods in Enzymology (Ginsburg, V., Ed.), Vol. 28, pp. 63-73, Academic Press, New York). In both glycopeptides and oligosaccharides, approximately one less residue of Man than predicted was determined. Both AE-PAD and liquid chromatographic analysis of borate-monosaccharide complexes with fluorometric detection (Mikami, H., and Ishida, Y. (1983) Bunseki Kagaku 32, E207-E210) gave similar quantification of mannose and other sugars. The capability of rapid, sensitive quantification of underivatized monosaccharides should facilitate structural analysis of glycoconjugates.