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.