薄层色谱-苯酚-硫酸法分析大豆低聚糖中棉籽糖含量  

Combination of Thin-layer Chromatography with Phenol-sulfuric Acid Method for Analyzing the Content of Raffinose in Soybean Oligosaccharide

作　　者：廖春龙[1,2] 邱奇琦[1,2] 印遇龙[1,2,3] 阮征[1,2] 

机构地区：[1]南昌大学食品科学与技术国家重点实验室,江西南昌330047 [2]南昌大学生命科学与食品工程学院,江西南昌330031 [3]中国科学院亚热带农业生态研究所,湖南长沙410125

出　　处：《食品科学》2010年第16期200-203,共4页Food Science

基　　金：南昌大学"赣江学者奖励计划"项目;中国博士后科学基金资助项目(20080440166)

摘　　要：目的:采用比色法建立一种适用于大豆低聚糖中棉籽糖快速检测的方法。方法:运用薄层层析技术分离单糖与低聚糖,展开剂为V(乙腈):V(冰乙酸):V(水)=6:3:2,显色剂为苯胺-二苯胺-磷酸,点样量为5μL,层析显色后,苯酚-硫酸法比色测定。结果:薄层层析可将大豆低聚糖中蔗糖、棉籽糖和水苏糖等组分分开,通过苯酚-硫酸法分析其中的棉籽糖,线性范围为20～80mg/mL(R2=0.9932),精密度RSD为1.56%,平均加样回收率为98.14%,RSD为3.99%。结论:以薄层层析-苯酚-硫酸法检测食品或大豆低聚糖中棉籽糖含量,测定结果与HPLC法对照,相对误差为3.22%,可用于大豆低聚糖中棉籽糖的检测。Objective： To establish a method for the fast determination of raffinose in food and oligosaccharide by the combined use of thin layer chromatography （TLC） and phenol-sulfuric acid method. Methods： For the separation of raffinose, the developing solvent used was a mixture of methyl cyanides, acetic acid and water （6：3：2, V/V）, and the chromogenic agent was composed of aminobenzene, diphenylamine and phosphoric acid, and the sample loading volume was 5 μL. The separated raffinose brand was harvested and analyzed by phenol-sulfuric acid method. Results： Different components of soybean oligosaccharide （sucrose, raffinose and stachyose） could be well separated by using TLC. The standard curve developed for raffinose determination by phenol-sulfuric acid method presented a linear range from 20 to 80 mg/mL （R2 = 0.9932）. The precision RSD for raffinose determination was 1.56%. The method spike recovery was 98.14%, with a RSD of 3.99%. The relative error between the results of determination by this method and HPLC was 3.22%. This method is applicable to the determination of raffinose in food and oligosaccharide.

关 键 词：大豆低聚糖 棉籽糖 薄层层析 苯酚-硫酸法 

分 类 号：TS247[轻工技术与工程—制糖工程]