响应面法优化高效液相色谱分析蜂蜜中5种生物胺的衍生条件

为确定高效液相色谱测定蜂蜜中5种生物胺(腐胺、精胺、尸胺、亚精胺、组胺)的最佳衍生化条件,在单因素优化基础上,采用响应面设计对衍生体系pH、衍生温度和时间等参数进行了优化,建立了5种生物胺的峰面积响应面模型。结果表明,建立的响应面模型拟合良好,各个模型的校正R2均大于0.99,响应预测值与实际值吻合度高,预测R2均大于0.99。3个衍生条件均能明显影响生物胺峰面积响应,影响大小依次为pH、衍生温度、衍生时间;pH和衍生时间存在显著交互作用(P<0.05)。最优的衍生条件为pH11、衍生温度45 ℃、衍生时间40 min,在此条件下衍生,5种生物胺的峰面积响应较好。使用两个浓度的生物胺溶液进行的验证实验结果与预测值相对偏差数值在2.32%~4.94%之间。

Optimization of Derivatization Conditions of Five Biogenic Amines in Honey by High Performance Liquid Chromatography with Response Surface Methodology

To optimize the derivatization conditions for determination of five kinds of biogenic amines(putrescine,spermine,cadaverine,spermidine,and histamine)in honey by high performance liquid chromatography,the conditions of pH,temperature and time of derivatization reaction were optimized by response surface design on the basis of single factor experiment. The results showed that the response models of five kinds of biogenic amines fitted well with adjusted R2>0.99. The predicted value was highly consistent with the actual response values,and all predicted R2>0.99. The influence of the three derivatization factors on biogenic amines peak response was pH,temperature and time in turn. There was significant interaction between pH and derivation time(P<0.05). The optimal conditions for derivatization reaction were pH11,45 ℃ and 40 min. With these parameters,the derivatization effect of the five biogenic amines was better. The relative deviation between the experimental values and the predicted values varied from 2.32% to 4.94% at two biogenic amines concentrations.