An Improved PMP Derivatization Method for Analyzing Monosaccharide Composition

Chem. J. Chinese Universities

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An Improved PMP Derivatization Method for Analyzing Monosaccharide Composition

LIN Xue¹, WANG Zhong-Fu¹, HUANG Lin-Juan², BAI Quan¹, JIA Jing-Fen¹

1. Life Science College, Key Laboratory of Ministry of Education for Western China Resource Biology and Biotechnology;
2. Institute of Modern Separation Science, Northwest University, Xi'an 710069, China

Received:2005-12-02 Revised:1900-01-01 Online:2006-08-10 Published:2006-08-10
Contact: WANG Zhong-Fu

Abstract

Abstract: An improved carbohydrate derivatization method employing 1-phenyl-3-methyl-5-pyrazolone(PMP) was developed in this study. Liquid ammonia was used instead of sodium hydroxide aqueous solution to provide the base medium for the derivatization reaction. Derivatization reaction and HPLC separation conditions were optimized, including reaction time, reaction temperature and PMP consumption. The derivatives of five reducing monosaccharides were well separated by HPLC. Because we employed ammonia instead of sodium hydro-xide, the excessive volatile ammonia can be removed by vacumm drying, these derivatives could be directly analyzed by MALDI-TOF-MS without desalting procedure, possessing potential application for monosaccharide and oligosaccharide separation and subsequent MS analysis. This method not only increased the MS detection sensitivity, but also decreased sample wastes during purification. We successfully applied this method to mo-nosaccharide composition analysis of polysaccharides SPPA-1, SPPB-1 and SPPC-1 purified from Spirulina Platensis.

Key words: HPLC, 1-Phenyl-3-methyl-5-pyrazolone, Derivatization, Saccharide

CLC Number:

O658

TrendMD:

LIN Xue¹, WANG Zhong-Fu¹, HUANG Lin-Juan², BAI Quan¹, JIA Jing-Fen¹. An Improved PMP Derivatization Method for Analyzing Monosaccharide Composition[J]. Chem. J. Chinese Universities, doi: .

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An Improved PMP Derivatization Method for Analyzing Monosaccharide Composition

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