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目的建立高效液相色谱-蒸发光散射检测(HPLC-ELSD)法,快速洗脱分离出蔗果三糖、蔗果四糖、蔗果五糖,测定益生菌粉中低聚果糖的含量。方法样品经过前处理,以乙腈和水为流动相,等度洗脱,经XAmide(4.6 mm×250 mm,5μm)中性酰胺色谱柱分离,采用蒸发激光检测器检测。结果蔗果三糖、蔗果四糖、蔗果五糖线性范围分别在0.2671~3.168 mg/m L、0.2429~2.8215 mg/m L、0.1617~3.544 mg/m L,线性相关系数r>0.99。3种主要组成的回收率在97.0%~105.0%之间。结论该方法可有效分离出蔗果三糖、蔗果四糖、蔗果五糖,推广性高、快速、简便、准确,可用于益生菌粉中低聚果糖含量的测定。
OBJECTIVE To establish a high performance liquid chromatography-evaporative light scattering (HPLC-ELSD) method for rapid determination of fructo-oligosaccharides in probiotics by HPLC. Methods The sample was pretreated with acetonitrile and water as mobile phases and eluted with isocratic elution. The sample was separated on a XAmide (4.6 mm × 250 mm, 5 μm) neutral amide column using an evaporative laser detector. Results The linear ranges of cane trisaccharide, cane fructotetraose and cane sugar were 0.2671 ~ 3.168 mg / m L, 0.2429 ~ 2.8215 mg / m L and 0.1617 ~ 3.544 mg / m L, respectively. The linear correlation coefficient r was 0.99. The recoveries of the three main components ranged from 97.0% to 105.0%. Conclusion The method can effectively separate sugarcane trisaccharide, sugarcane tetraose and sugarcane pentasaccharide. The method is rapid, simple, accurate and can be used for the determination of fructooligosaccharides in probiotic powder.