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.