恰玛古多糖含量测定方法的优化

Optimization of the method for the determination of polysaccharide content in Brassica rapa L.

中文关键词:  恰玛古,多糖,苯酚-硫酸法,3, 5-二硝基水杨酸(DNS)法

英文关键词:Brassica rapa L.  polysaccharide  sulfuric acid-phenol method  3, 5-dinitrosalicylic acid method(DNS)

中文摘要:

      恰玛古在新疆以卓越的药用价值著称,其多糖成分因显著的生物学效应成为科研焦点,引起了广泛而深入的研究热潮。目前较多研究中恰玛古多糖含量依旧利用苯酚-硫酸法进行测定,但是传统苯酚-硫酸法不能排除样品中的可能直接与硫酸显色的还原性糖和非多糖杂质的干扰,为了克服以上缺点,得到真实的恰玛古多糖含量,本研究将传统的苯酚-硫酸法和3, 5-二硝基水杨酸(DNS)法相结合,建立了恰玛古多糖含量测定的方法。我们采用 Box-Behnken 实验设计的响应面工艺,筛选出测定过程中的最佳条件组合:总糖测定时,反应时间设为27.22 min,苯酚试剂与硫酸的用量分别为0.32 mL和1.08 mL；而在还原糖测定中,显色反应时间优化为20.53 min,DNS显色剂用量为2.08 mL,并需静置14.90 min以确保充分反应。利用上述确定的最佳条件测定的恰玛古多糖含量为36.4732 g/100g。通过精密度、重复性、稳定性实验对测量结果进行考察表明,苯酚-硫酸法和3, 5-二硝基水杨酸(DNS)组合方法不仅有效解决了传统方法中的干扰问题,还确保了测量结果的可靠性与有效性,为恰玛古多糖的定量研究提供了坚实的技术支撑。本研究的成功实施,不仅弥补了恰玛古多糖含量测定领域的空白,也为相关科研工作者提供了科学、准确的测定方法,有望推动恰玛古多糖研究领域的深入发展。

英文摘要:

      Brassica rapa L. is renowned in Xinjiang for its outstanding medicinal value, and its polysaccharide components have become the focus of scientific research due to their remarkable biological effects, triggering a wave of extensive and profound research. At present, most of the investigations still adopt the sulfuric acid-phenol method for the determination of polysaccharide content in Brassica rapa L. Still, the traditional phenol-sulfuric acid method cannot exclude the interference of reducing sugars and non-polysaccharide impurities in the samples, which may be directly chromatographed with sulfuric acid. To overcome the above shortcomings and obtain the real polysaccharide content of Brassica rapa L., this study combined the traditional sulfuric acid-phenol method and 3,5-dinitrosalicylic acid (DNS) method to establish an approach for the determination of the polysaccharide content. We used the response surface process of the Box-Behnken experimental design to screen out the optimal conditions affecting the determination factors. The optimal conditions in the total sugar content determination method were a reaction time of 27.22 min; a phenol reagent dosage of 0.32 mL; and a sulfuric acid dosage of 1.08 mL; The optimal conditions for the determination of reducing sugars were 20.53 min for the color development reaction time; 2.08 mL for the color developer DNS; and 14.90 min for the resting time. Through rigorous precision, repeatability, and stability experiments, the assessment of measurement results demonstrates that the combined method of sulfuric acid-phenol and 3, 5-dinitrosalicylic acid (DNS) not only effectively addresses the interference issues inherent in traditional methods but also ensures the reliability and validity of measurement outcomes. This provides a solid technical foundation for the quantitative study of Brassica rapa L. polysaccharide content. The successful implementation of this study not only fills the gap in the field of Brassica rapa L. polysaccharide content determination but also offers scientific and accurate measurement methods to relevant researchers, potentially fostering further advancements in the research of Brassica rapa L. polysaccharides.