高效液相色谱法测定壳寡糖的含量
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摘要
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 NH_2(4.6 mm×250 mm,5 μm), 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(r~2) 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.
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 NH_2(4.6 mm×250 mm,5 μm), 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(r~2) 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.
引文
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[10]邵波,陈梅兰,金月珠.离子色谱快速检测壳寡糖的含量[J].药物分析杂志,2017,37(11):1 973-1 977.
[11]马昊,卢珩俊,郑后涛,等.五种低聚壳寡糖阴离子交换色谱-积分脉冲安培法测定[J].浙江树人大学学报,2016,16(2):11-15.
[12]李娟,朱捷,李春红,等.氨基寡糖素的高效阴离子交换-脉冲安培检测法[J].食品科学,2015(21):295-298.
[13]王礼,邓卫利,林文业.壳寡糖分析检测的研究进展[J].大众科技,2011,33(5):109-110.
[14]杨菊林,韩宝芹,刘万顺.酶促反应制备壳寡糖及壳寡糖分析[J].实验与技术,2008,32(6):5-8.
[15]蒋慧,李恒,张澜,等.高效液相色谱法测定壳寡糖的含量[J].发酵科技通讯,2016,45(4):232-236.
[2]李苏亮,闫冰雪,郑素霞.壳寡糖的理化特性及制备方法[J].添加剂世界,2016(7):27-29.
[3]陈虹,侯伟革.壳寡糖及其应用[J].饲料博览,2007(21):23-24.
[4]陈海燕,张彬,何勇松.壳寡糖的研究进展和应用前景[J].广东畜牧兽医科技,2007,32(2):17-20
[5]KANG Y R,CHOI H Y.Effect of supplementation of lowmolecular-weight chitosan oligosaccharide,GO2KA1,on postprandial blood glucose levels in healthy individuals following bread consumption[J].Food Science and Biotechnology,2016,25(3):911-914.
[6]徐泽平,杨传伦,张心青,等.乙酰丙酮测定壳寡糖的含量[J].现代科学仪器,2012,6(3):64-67.
[7]谭佩毅,黄秀锦.Imoto比色法测定壳寡糖含量的研究[J].食品工业科技,2012,33(22):71-74.
[8]王晓燕,许牡丹.一种新型壳低聚糖检测方法[J].食品工业科技,2008,29(4):271-273.
[9]孟显丽,陈国华,孙明昆,等.薄层色谱法分析壳寡糖[J].青岛海洋大学学报,2002,32(4):641-644.
[10]邵波,陈梅兰,金月珠.离子色谱快速检测壳寡糖的含量[J].药物分析杂志,2017,37(11):1 973-1 977.
[11]马昊,卢珩俊,郑后涛,等.五种低聚壳寡糖阴离子交换色谱-积分脉冲安培法测定[J].浙江树人大学学报,2016,16(2):11-15.
[12]李娟,朱捷,李春红,等.氨基寡糖素的高效阴离子交换-脉冲安培检测法[J].食品科学,2015(21):295-298.
[13]王礼,邓卫利,林文业.壳寡糖分析检测的研究进展[J].大众科技,2011,33(5):109-110.
[14]杨菊林,韩宝芹,刘万顺.酶促反应制备壳寡糖及壳寡糖分析[J].实验与技术,2008,32(6):5-8.
[15]蒋慧,李恒,张澜,等.高效液相色谱法测定壳寡糖的含量[J].发酵科技通讯,2016,45(4):232-236.