二次回归通用旋转组合设计法优化枸杞多糖的荧光标记  

Using quadratic regression universal rotary combination design to optimize the fluorescence labeling condition of lycium barbarum polysaccharide

作　　者：蔡慧珍[1] 吴慧珊[1] 杨晓辉[1] 任彬彬[1] 

机构地区：[1]宁夏医科大学公共卫生学院,银川750004

出　　处：《国际中医中药杂志》2016年第4期355-358,共4页International Journal of Traditional Chinese Medicine

基　　金：2013年度宁夏高等学校科学研究项目（NGY2013072）

摘　　要：目的 利用二次通用旋转组合设计试验筛选枸杞多糖荧光标记的最优标记条件。方法 枸杞多糖与酪胺氨基共价偶联后与异硫氰酸荧光素反应，实现选择性荧光标记。利用二次通用旋转组合设计试验，研究缓冲液pH值、标记时间、温度、酪胺量与标记效率之间的关系，从中筛选出最佳的标记条件。结果 得回归方程为：Y＝0.085 41－0.002 82 X1－0.015 68 X2＋0.008 11 X3＋0.005 01 X4＋0.008 75 X1X2－0.005 75 X1X3－0.001 75 X1X4＋0.010 63 X2X3＋0.000 125 X2X4＋0.000 25 X3X4－0.021 44 X12－0.008 89 X22－0.001 984 X32＋0.003 66 X42，失拟性检验F＝1.76＜F0.05（5,3）＝9.01；显著性检验F＝8.42＞F0.01（11,8）＝5.74，表明二次回归模型有统计学意义。枸杞多糖荧光标记时温度在50 ℃，pH值为8.5，反应时间设为4 d时结合效率最高。结论 在枸杞多糖荧光标记的过程中，在特定的反应温度和缓冲液pH值条件下，适当延长反应时间可提高反应效率。Objective Quadratic regression universal rotary combination design was used to optimize the fluorescence labeling condition of lycium barbarum polysaccharide （LBP）. Methods The fluorescence labeling condition of LBP presented, when the covalent coupling of LBP and tyramine was reacted with fluorescein isothiocyanate （FITC）. Filter the best labeling condition via using quadratic regression universal rotary combination design experiment on the relationship of labeling efficiency among pH value of buffer solution, reaction time, temperature and the dose of tyramine. Results The regression equation was： Y=0.085 41 - 0.002 82X1 - 0.015 68X2 ＋ 0.008 11 X3 ＋ 0.005 01 X4 ＋ 0.008 75 X1X2 - 0.005 75 X1X3 - 0.001 75 X1X4 ＋ 0.010 63 X2X3 ＋ 0.000 125 X2X4 ＋ 0.000 25 X3X4 - 0.021 44 X12 - 0.008 89 X22 - 0.001 984 X32 ＋ 0.003 66 X42, and the variables from X1 to X4 represented pH value, reaction time, temperature and dose of tyramine, respectively. The goodness of fitting of regression equation was statistically significant. The condition of labling LBP was optimized when the temperature was at 50 ℃, timing of labling at the fourth day and pH value was 8.5. Conclusion The condition of labling LBP was optimized with suitable temperature, pH value and extended timing.

关 键 词：枸杞多糖 异硫氰酸荧光素 酪胺 二次通用旋转组合设计 

分 类 号：R284[医药卫生—中药学]