HPLC-ELSD测定壳寡糖低聚木糖胶囊中的低聚木糖

Quantification of Xylooligosaccharides in Chitooligosaccharide Xylooligosaccharide Capsules by HPLC-ELSD

目的建立酰胺基亲水作用色谱的高效液相色谱-蒸发光散射法(HPLC-ELSD)测定壳寡糖低聚木糖胶囊中的低聚木糖。方法选用Waters XBridge Amide(4.6 mm×250 mm,3.5μm)色谱柱,以乙腈-0.2%三乙胺为流动相进行梯度洗脱,流速1.0 m L·min^(-1),进样量20μL,柱温45℃。结果对照品和样品溶液中木糖和低聚木糖7个组分得到很好的分离,各组分在0.05~0.5 mg·m L^(-1)内线性良好,相关系数r=0.999 4~0.999 9;7个组分在高低水平的添加回收率(n=6)为90.6%~103.2%,RSD<4.0%;3批样品中低聚木糖含量X2~X7分别为313,312,310 mg·g^(-1),X2~X4分别为282,279,278 mg·g^(-1)。结论本方法操作简单快速、灵敏度高、重复性好,为低聚木糖制品的质量控制及生产工艺研究提供更准确可靠的新方法。

OBJECTIVE To establish an HPLC-ELSD method for the determination of xylooligosaccharide in chitooligosaccharide xylooligosaccharide capsules by amide hydrophilic interaction chromatography. METHODS The chromatographic separation was achieved on a Waters XBridge Amide column（4.6 mm×250 mm, 3.5 μm） by a gradient elution program with acetonitrile-0.2% triethylamine as the mobile phase. The flow rate was 1.0 m L·min-1, injection volume was 20 μL, column temperature was 45 ℃. RESULTS The calibration curves of xylose and xylooligosaccharide in the mixed reference substances or sample solutions were well separated with good correlation coefficient（r=0.999 4-0.999 9） in the concentration range of 0.05-0.5 mg·m L^（-1）. The recoveries（n=6） of 7 components at high and low concentration were 90.6%-103.2%, the RSDs were 4.0%. The contents of xylooligosaccharide（X2-X7） in three batches were 313, 312, 310 mg·g^（-1）, and xylooligosaccharide（X2-X4） 282, 279, 278 mg·g^（-1）, respectively. CONCLUSION The method is simple, rapid, sensitive and reproducible, providing a more accurate and reliable method for the quality control and research of production process in xylooligosaccharide products.