期刊
ANALYTICAL BIOCHEMISTRY
卷 368, 期 1, 页码 79-86出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2007.05.016
关键词
keratan sulfate; heparan sulfate; dermatan sulfate; LC/MS/MS; mucopolysaccharidosis; plasma; serum
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.
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