高效液相色谱法测定壳寡糖的含量

Determination of chitosan oligosaccharide by high performance liquid chromatography

建立高效液相色谱检测壳寡糖的方法。壳寡糖经浓盐酸水解生成盐酸氨基葡萄糖,以高效液相色谱法测定盐酸氨基葡萄糖来确定壳寡糖的含量。称取0.1 g壳寡糖样品,加浓盐酸5 mL于(100±1)℃水浴中水解3 h。选用氨基柱(4.6 mm×250 mm,5μm),流动相为乙腈–水(体积比75∶25),流量为1.0 mL/min,采用内部温度为35℃的示差检测器进行检测。盐酸氨基葡萄糖的质量浓度在1.0～8.0 mg/mL范围内与色谱峰峰面积呈良好的线性关系,相关系数(r^2)为0.998 9,方法检出限为0.8%。测定结果的相对标准偏差为1.5%(n=6),加标回收率为97.4%～99.2%。方法操作简单、定量准确,适合于壳寡糖的检测。

A high performance liquid chromatography method for determination of chitosan oligosaccharide was developed.Chitosan oligosaccharides were hydrolyzed by concentrated hydrochloric acid to form glucosamine hydrochloride,and the content of chitosan oligosaccharides was determined by high performance liquid chromatography with glucosamine hydrochloride.After 0.1 g chitosan oligosaccharide sample being weighed,5 mL of concentrated hydrochloric acid was added to hydrolyze for 3 h in water bath with the temperature of(100±1)℃.The analysis column was NH2(4.6 mm×250 mm,5μm),and the mobile phase was acetonitrile–water solution(volume ratio was 75∶25)with the flow rate of 1.0 mL/min.The detector was differential refractometer with the internal temperature of 35℃.The mass concentration of glucosamine hydrochloride was linear with the peak area of the chromatograpy in the range of 1.0–8.0 mg/mL.The correlation coefficient(r2)was 0.998 9 and the detection limit was 0.8%.The relative standard deviation of determination results was 1.5%(n=6).The recoveries ranged from 97.4%to 99.2%.The method is simple and accurate,it is suitable for the detection of chitosan oligosaccharides.