Analysis of monosaccharide composition of water‐soluble polysaccharides from Codium fragile by ultra‐performance liquid chromatography‐tandem mass spectrometry

Codium fragile is a green alga belonging to Codiales family. The sulfated polysaccharides of this alga have anti-coagulation, antiviral, anti-angiogenesis, antioxidant, and immunoregulatory properties. In this study, we developed a reliable and rapid method for the analysis of 10 monosaccharides using ultra-performance liquid chromatography-tandem mass spectrometry in the negative electrospray ionization and multiple reaction monitoring mode. Monosaccharides, including two pentoses (xylose, arabinose); two deoxyhexoses (rhamnose, fucose); three hexoses (mannose, glucose, galactose); two hexuronic acids (glucuronic acid, galacturonic acid), and an N-acetyl-hexosamine (glucosamine), were derivatized using 1-phenyl-3-methyl-5-pyrazolone and simultaneously analyzed within 9 min. Optimization of the derivatization process, especially by using various 1-phenyl-3-methyl-5-pyrazolone concentrations, was studied. The calibration curves showed good linearity with a squared correlation coefficient > 0.995. The spiked recovery was determined to be 91.1-105.7% with the relative intra-day and inter-day standard deviations ranging from 2.58-6.71% and 3.15-7.67%, respectively. The limit of detection and limit of quantification for all 10 monosaccharides ranged from 0.02 to 0.10 μg/mL and 0.05 to 0.25 μg/mL, respectively. Using this method, the monosaccharides comprising the polysaccharides of Codium fragile were determined to be arabinose, galactose, and glucose.