采用紫外可见分光光度法研究了刚果红浓度、β-葡聚糖浓度、反应温度、pH值、反应时间、离子浓度、光照、体系中可能共存的淀粉和蛋白等因素对测定燕麦β-葡聚糖含量的影响。同时,在紫外吸收光谱分析的基础上运用化学计量学方法计算刚果红与燕麦β-葡聚糖的结合常数K与结合位点数,N及平均结合数n。结果表明:室温条件下,刚果红质量浓度50μg/mL,pH7.5,反应时间30min,离子浓度0.1mol/L时,吸光度与β-葡聚糖浓度在0~10μg/mL范围内具有较好的线性关系。光照对测定无影响,共存的淀粉和蛋白质对测定影响较小。在试验条件下,刚果红与燕麦β-葡聚糖的结合常数K为4.08×10^6、结合位点数N为336、平均结合数n为297。
Spectrophotometry was used to determine the content of oat β - glucan. The effects of some factors were investigated, including the concentration of Congo red and oat β - glucan, reaction temperature, pH, reaction time, ion concentration, exposure to light, and the starch and protein coexisting in the system. The binding equilibri- um constant K, the total number of binding sites per β - glucan molecule N, and the binding number of Congo red per β- glucan molecule n were determined by chemometrics based on the analysis of ultraviolet - visible absorption spectra. The results showed that the absorbency had a linear relationship with β -glucan in the concentration from 0 to 10 μg/mL under the condition that room temperature, Congo red concentration 50 μg/mL, pH 7.5, reaction for 30 min, and ion concentration 0.1 mol/L. Exposure to light had no effect on the determination. Starch and protein had a relatively low effect on the absorbency. The binding equilibrium constant K, binding sites N and average bind- ing number n are 4.08 × 10^6, 336 and 297, respectively under the experimental conditions.