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测定麦芽糖转糖苷反应体系组成的高效液相色谱法

Determination of Composition of Transglycosidation System for Maltose by High Performance Liquid Chromatography

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【作者】 邹耀洪居红芳陈爱英

【Author】 ZOU Yao _ hong, JU Hong _ fang, CHEN Ai _ ying(Department of Biochemistry, Changshu College, Changshu 215500, China)

【机构】 常熟高等专科学校生物化学系常熟高等专科学校生物化学系 江苏常熟215500江苏常熟215500江苏常熟215500

【摘要】 建立了分析低聚异麦芽糖组成的高效液相色谱法 ,采用Spherisorb_NH2色谱柱,示差折光检测器 ,乙腈 -水 (体积比70∶30)为流动相 ,外标法定量测定 ;结果显示各糖质量浓度在0.1~10g/L范围内与峰面积呈良好线性关系 ,相关系数为0.9990~0.9997 ;应用该法跟踪了pH5.0的柠檬酸 -柠檬酸钠缓冲溶液中以α_葡萄糖转苷酶为催化剂在58℃温度下的麦芽糖转糖苷反应 ,分析了反应体系组成随时间的变化 ,得到了上述反应条件下麦芽糖最大限度地转化为低聚异麦芽糖的最佳反应时间为24h;该法快捷、简单、准确 ,可用于低聚异麦芽糖生产的质量控制。

【Abstract】 A new method has been developed for analysing the composition of isomaltooligosaccharide by high performance liquid chromatography with a column of Spherisorb _ NH2 300 mm×4.6 mm(id) and differential refractometer using acetonitrile-water (volume ratio 70∶30)as a mobile phase. The carbohydrates were quantified by external standard method. The results show that there are a good linear relation of peak area to mass concentration of the carbohydrates in the range of 0.1~10 g/L with correlation coefficients range from 0.999 0 to 0.999 7. The method has been used to trace the transglycosidation for maltose with α _ glucosyltransferase as a catalyst at 58 ℃ in citric acid-sodium citrate buffer of pH 5.0. The results obtained by analysing the composition of transglycosidation system for maltose under the temperature show that optimum reaction time of maltose transforming into isomaltooligosaccharide is 24 h. The method is rapid, simple and accurate, and can be used for quality control of isomaltooligosaccharide production.

【基金】 江苏省教育厅自然科学基金项目(01KJD150006)
  • 【文献出处】 分析测试学报 ,Journal of Instrumental Analysis , 编辑部邮箱 ,2002年03期
  • 【分类号】O657.7
  • 【被引频次】9
  • 【下载频次】145
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