| Three high performance liquid chromatography methods for the determination of polysaccharide composition of oil-tea camellia seed cake were put up and compared, which were NH2 column- refractive index detector (RID) method, NH2 column- tandem mass spectrometry (MS/MS) method, 1-phenyl-3-methyl-5-pyrazolone (PMP) pre-column derivation- C18 column- ultra-violet detector (UV) method (pre-column derivation method), respectively. Considering the separation effect, sensitivity, instrument cost and other factors, the pre-column derivation method was selected.The method could be used for determination of rhamnose, mannose, glucose, galactose, xylose, arabinose, glucuronic acid and galacturonic acid simultaneously. The correlation coefficients were higher than 0.99, with limits of detection 0.02-0.29 mg/L and limits of quantification 0.06-0.96 mg/L.The recoveries of standard addition of monosaccharides and uronic acids were 77.2%-111.9% and 60.1%-87.0% respectively. Intra-day relative standard deviations (RSD) of the peak area were 0.08%-377% while inter-day RSD were 0.39%-17.15%.The established method was then applied to detect four polysaccharide samples of oil-tea camellia seed cake. The results showed that all the polysaccharide samples consisted of five or six monosaccharides and two uronic acids, and glucose content was the highest. |
