Analytical method for the determination of disaccharides derived from keratan, heparan, and dermatan sulfates in human serum and plasma by high-performance liquid chromatography/turbo ionspray ionization tandem mass spectrometry
Analytical method for the determination of disaccharides derived from keratan, heparan, and dermatan sulfates in human serum and plasma by high-performance liquid chromatography/turbo ionspray ionization tandem mass spectrometry
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DOI：
10.1016/j.ab.2007.05.016
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发表时间：
2007-09-01
影响因子：
2.9
通讯作者：
Okazaki, Osamu
中科院分区：
文献类型：
--
作者：
Oguma, Toshihiro;Tomatsu, Shunji;Okazaki, Osamu
建立了高灵敏的液质联用(LC-MS/MS)分析硫酸角蛋白(KS)、硫酸乙酰肝素(HS)和硫酸皮肤素(DS)产生的双糖的方法。结果表明，人血清和血浆中的KS、HS和DS分别被角聚糖酶11、肝素酶和软骨素酶13消化为各自的二糖。双糖的分析采用LC-MS/MS，在负离子模式下进行多反应监测。LC分离在Hypercarb(2.0 mm i.d.)上进行。×150 mm，5微米)，乙腈-0.01M碳酸氢铵(PH 10)梯度洗脱。流动相流速为0.2ml/min。用配备涡轮喷雾的API-4000型质谱仪测定了正常人血清和粘液多糖(NIPS)患者血浆中各糖胺多糖(GAG)的含量。以变异系数表示的日内精密度在3个人对照样本的5次重复分析中在15.80/0以内。粘粒间(总体n=15)3d精密度在14.8%以内。该方法灵敏、重复性好，可用于临床诊断。(C)2007 Elsevier Inc.保留所有权利。
We established a highly sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method to analyze the disaccharides produced from keratan sulfate (KS), heparan sulfate (HS), and dermatan sulfate (DS). It was revealed that KS, HS, and DS in human serum and plasma were digested to each disaccharide by keratanase 11, heparitinase, and chondroitinase 13, respectively. Analysis of disaccharides was performed by LC-MS/MS with multiple reactions monitoring in the negative ion mode. Separation of LC was performed on a Hypercarb (2.0 mm i.d. x 150 mm, 5 mu m) with a gradient elution of acetonitrile-0.01 M ammonium bicarbonate (pH 10). The mobile phase flow rate was 0.2 ml/min. An API-4000 mass spectrometer equipped with a turbo ionspray was used to determine each glycosaminoglycan (GAG) in the serum of control subjects and plasma of muco polysaccharidose (NIPS) patients. The intraday precision expressed as a coefficient of variation was within 15.80/0 for five replicate analyses with three human control samples. The interclay (overall, n = 15) precision was within 14.8% for 3 days. This method is sensitive and reproducible, and it would be useful for clinical diagnosis. (c) 2007 Elsevier Inc. All rights reserved.