高效液相法测定苦槠淀粉多糖的单糖和双糖组成

Determination of monosaccharides and disaccharides in Castanopsis sclerophylla starch by high performance liquid chromatography

采用高效液相色谱法测定苦槠淀粉多糖的单糖和双糖组成。以玉米淀粉等作为对照,采用三氟乙酸水解淀粉多糖,水解产物使用高效液相色谱-蒸发光散射法(HPLC-ELSD)检测,以7种糖标准品为对照,测定水解产物中单糖和双糖的组成。结果表明,当流动相为乙腈-0.02%氨水(75:25),柱温40℃,雾化温度30℃,漂移管温度70℃,空气流量为2.5 m L/min时,7种糖标准品在(0.0047~4.28)mg/m L范围内线性关系良好,最低检测限介于(0.789~25.048)ng/m L,稳定性和精密度的偏差依次不高于2.89%、1.67%。采用该方法具有准确度高、分离效果明显、稳定性强等优点,测得苦槠淀粉中鼠李糖含量为5.55 mg/g、果糖含量为3.08 mg/g、葡萄糖含量为7.31 mg/g、麦芽糖含量为15.94 mg/g。

To analyst monosaccharide and disaccharides composition in Castanopsis sclerophylla starch by high performace liquid chromatography （HPLC）. The hydrolysis of Castanopsis sclerophylla starch, corn starch, potato starch and wheat starch by trifluoroacetic acid （TFA）, the monosaccharide and disaccharides composition was assayed directly by high performance liquid chromatography-evaporative light-scattering detector （HPLC-ELSD） with seven monosaccharide and disaccharides as control. The results showed that using XBridge Amide 5 lam chromatographic column at 40℃, the ratio of acetonitrile and 0.02% ammonium hydroxide was 75：25 for isocratic elution with the flow rate of 0.5 mL/min. The nebulization chamber temperature and the drift tube temperature were 30 ℃and 70 ℃ in turn, and air flow rate was 2.5 mL/min. The seven monosaccharides and disaccharides concentration in the range of（0.0047-4.28）mg/mL showed good linearity with their peak areas. The standard deviation of precision and stability were not more than 1.67% and 2.89% respectively, and the limit of detention was （0.789-25.048） ng/mL. The HPLC-ELSD method could be applied to analysis the contents of monosaccharides and disaccharides from Castanopsis sclerophylla starch, and the rhamnose, fructose, glucose, maltose contents respectively were 5.55, 3.08, 7.31, 15.94 mg/g.