食品科学 ›› 2010, Vol. 31 ›› Issue (16): 200-203.doi: 10.7506/spkx1002-6630-201016043

• 分析检测 • 上一篇    下一篇

薄层色谱-苯酚硫酸法分析大豆低聚糖中棉籽糖含量

廖春龙1,2,邱奇琦1,2,印遇龙1,2,3,阮 征1,2,*   

  1. 1.南昌大学 食品科学与技术国家重点实验室 2.南昌大学生命科学与食品工程学院 3.中国科学院亚热带农业生态研究所
  • 收稿日期:2009-11-24 修回日期:2010-05-03 出版日期:2010-08-25 发布日期:2010-12-29
  • 通讯作者: 阮征 E-mail:ezruan@yahoo.com

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

Zheng RUAN   

  • Received:2009-11-24 Revised:2010-05-03 Online:2010-08-25 Published:2010-12-29
  • Contact: Zheng RUAN E-mail:ezruan@yahoo.com

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。

关键词: 大豆低聚糖, 棉籽糖, 薄层层析, 苯酚- 硫酸法

Abstract:

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.

Key words: soybean oligosaccharide, raffinose, TLC, phenol-sulfuric acid method

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