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建立了一种同时测定岩藻多糖7种单糖及2种糖醛酸含量的柱前衍生-高效液相色谱检测方法。岩藻多糖用三氟乙酸水解、1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化,并采用配有紫外检测器的反相高效液相色谱仪在250nm波长下进行测定,实现了7种单糖和2种糖醛酸的良好分离。7种单糖和2种糖醛酸在1～50μg/mL范围内线性关系良好,相关系数r>0.998,回收率在86%～105%之间,相对标准偏差均<10%。甘露糖和岩藻糖检出限分别为0.5μg/mL和2.0μg/mL,其余单糖和糖醛酸检出限为1.0μg/mL。该方法重现性好,能够快速地对多糖样品进行组成分析。
A method was developed for the simultaneous determination of seven monosaccharides and two uronic acids in fucoidan by pre-column derivatization-high performance liquid chromatography (HPLC). Fucoidan was hydrolyzed with trifluoroacetic acid and derivatized with 1-phenyl-3-methyl-5-pyrazolone (PMP) using an RP-HPLC equipped with UV detector at 250 nm The measurement was carried out to achieve a good separation of seven monosaccharides and two uronic acids. The linearity of 7 kinds of monosaccharides and 2 kinds of uronic acid in the range of 1 ~ 50μg / mL was good with the correlation coefficient r> 0.998. The recoveries were between 86% and 105% with relative standard deviations <10%. The detection limits of mannose and fucose were 0.5μg / mL and 2.0μg / mL, respectively, and the limits of detection of other monosaccharides and uronic acids were 1.0μg / mL. The reproducibility of this method allows quick compositional analysis of polysaccharide samples.