RAPID QUANTITATIVE-DETERMINATION OF SIALIC ACIDS IN GLYCOPROTEINS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH A SENSITIVE FLUORESCENCE DETECTION
RAPID QUANTITATIVE-DETERMINATION OF SIALIC ACIDS IN GLYCOPROTEINS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH A SENSITIVE FLUORESCENCE DETECTION
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DOI：
10.1006/abio.1995.1432
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发表时间：
1995-09-01
影响因子：
2.9
通讯作者：
ANUMULA, KR
中科院分区：
文献类型：
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作者：
ANUMULA, KR
用邻苯二胺2 HCl（OPD）特异性标记唾液酸以产生稳定的荧光喹喔啉衍生物。在NaHSO 4溶液（0.25 M 80 ℃，20 min）中从糖蛋白释放唾液酸，并在相同溶液中用OPD（10 mg/ml（终浓度））衍生化。80 ℃，40 min），在C-18反相Ultrasphere-ODS柱上分离OPD衍生化的各种唾液酸，使用如前所述的用于测定邻氨基苯甲酸衍生化单糖的溶剂系统和检测器条件。常见的N-乙酰基-和N-羟乙酰基神经氨酸在20分钟内分离，其他N-和O-酰化唾液酸在40分钟内分离。和三酰化唾液酸在本分离中被分离成它们各自的基团。乙酰神经氨酸的激发波长为232 nm，发射波长为420 nm，
Sialic acids were specifically labeled with o-phenylenediamine 2HCl (OPD) to yield stable fluorescent quinoxaline derivatives. The sialic acids were released from the glycoprotein in a NaHSO4 solution (0.25 M 80 degrees C, 20 min) and derivatized in the same solution with the OPD (10 mg/ml (final cone.) 80 degrees C, 40 min), Various sialic acids derivatized with the OPD were separated on a C-18 reversed-phase Ultrasphere-ODS column using the solvent systems and the detector conditions used for the determination of monosaccharides derivatized with anthranilic acid as reported earlier. The common N-acetyl- and N-glycolylneuraminic acids were separated within 20 min, and the other N- and O-acylated sialic acids were separated in 40 min. The OPD derivatives of mono-, di-, and triacylated sialic acid were separated into their respective groups in the present separation The fluorescence maxims for the OPD-N-acetylneuraminic acid were 232 nm excitation and 420 nm emission and the limit of quantitation was