Résumé
Objective To analyze the monosaccharide composition and the relative content of different alcohol precipitations of Panax japonicus polysaccharides.Methods The four components of Panax japonicus polysaccharides were isolated by stepwise ethanol precipitation method.The four components of Panax japonicus polysaccharides were hydrolyzed with trifluoroacetic acid (TFA) and derivated by l-phenyl-3-methyl-5-pyrazolo (PMP),respectively.The monosaccharide composition and relative content were analyzed using LC-FT-ICR-MS and HPLC-UV method.Results The four components of Panax japonicus polysaccharides consisted of glucose,rhamnose,galacturonic acid,galactose,arabinose,and glucose was the main monosaccharide.With the increase of ethanol concentration,the relative content of Ara increased gradually,as while the GalA decreased.Conclusions Precolumn derivation HPLC method was successfully applied for the determination of the monosaccharides in Panax japonicus polysaccharides,and there were differences in the four ethanol precipitations of Panax japonicus polysaccharides.The study can provide a basis for the separation of Panax japonicus polysaccharides.
Résumé
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.