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Updated: Jan 31, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
[Analysis of monosaccharide composition of twelve polysaccharides by pre-column derivatization procedure UPLC-MS/MS]
Yu-Ting Liang1, Jun-Hui Zhou2, Tie-Gui Nan2
1School of Chinese Materia Medica, Capital Medical University, Beijing 100071, China.
Abstract:
This study is to establish a pre-column derivatization procedure with 1-phenyl-3-methyl-5-pyrazolone (PMP) UPLC-MS/MS method for the determination of the monosaccharide composition of 12 polysaccharides. At the same time, the monosaccharide components of polysaccharides in Armillaria gallica were analyzed. The separation was performed on a ACQUITY ZORBAX RRHD Eclipse Plus C₁₈ column(2.1 mm×100 mm, 1.8 μm),using 95% acetonitrile (A) and ammonium acetate-5% acetonitrile-water (B) as mobile phase with gradient elution. The target components were detected in multiple-reaction monitoring (MRM) mode by mass spectrometry with electrospray ionization (ESI) source operated in ionization mode. The results showed that based on the monosaccharides detection method established by UPLC-MS/MS, the linearity of the 12 monosaccharides components were linear in their linear range (R²>0.990), and the recovery rate were 92.30%-105.6%. 11 monosaccharides such as fructose, mannose, and glucose were detected in A. gallica samples. The method established in this experiment is robust, highly reproducible and accurate, and is suitable for the determination of monosaccharide components such as A. gallica.
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