液相色谱双柱法和高效离子交换色谱法测定低聚半乳糖含量比较

Determination of GOS by high performance ion exchange chromatography and double-HPLC-RI methods

目的:优化对比液相色谱双柱法和高效离子交换色谱法两种测定低聚半乳糖含量的方法,同时建立一种新的低聚半乳糖换算系数(K值)计算方法;方法:对现有上述两种测定低聚半乳糖含量的检测方法进行对比分析,筛选出分离效果更好的糖柱,并进一步优化色谱条件。结果:优化后的液相色谱双柱法对低聚半乳五糖到低聚半乳八糖的分离效果明显提升,提高了测定准确度,而高效离子交换色谱法操作更加简便;根据不同样品选用不同K值来计算低聚半乳糖含量的方式可进一步提升结果的精确性。结论:两种方法均可有效对低聚半乳糖含量进行检验分析,高效离子交换色谱法检测灵敏度较高,且已被AOAC等国际权威机构纳入标准检测方法,可作为低聚半乳糖含量测定的标准仲裁法;液相色谱双柱法测定低聚半乳糖更为简便快捷,经济适用,可用于生产企业日常产品检测。

Objective:Optimization of two methods for the determination of galacto-oligosaccharide by comparative liquid chromatography double column and high performance ion exchange chromatography,and development of a new method for the calculation of the conversion factor(K value)of galacto-oligosaccharide.Methods:The two existing assays for the determination of galacto-oligosaccharide content described above were compared and analysed to select a sugar column with better separation and to further optimize the chromatographic conditions.Results:The optimized dual-column liquid chromatography method has significantly improved the separation of oligogalactopentasaccharides to oligogalacto-octasaccharides,improving the accuracy of the determination,while the high performance ion exchange chromatography method is simpler to operate;the accuracy of the results can be further improved by using different K values for different samples to calculate the oligogalactose content.Conclusion:Both methods can effectively test and analyse the content of oligogalactose.The high performance ion exchange chromatography method is more sensitive and has been incorporated into the standard test methods by international authorities such as AOAC,so it can be used as a standard arbitration method for the determination of galactooligosaccharide content;the liquid chromatography double column method is simpler,faster and more economical for the determination of galacto-oligosaccharide and can be used for daily product testing by manufacturers.