Optimization of experimental variables in the dabsyl chloride derivatization of biogenic amines for their determination by RP-HPLC

In the last few years special attention has been paid to the pre-column der ivatization of biogenic amines with dabsyl chloride because proper experime ntal conditions for this reaction are very important. In this study, an exp erimental design (Doehlert design) was used to optimize the variables invol ved in the dabsylation of the following amines: histamine, tyramine, phenyl ethylamine, tryptamine, cadaverine, putrescine, spermidine, and spermine. T he optimum experimental conditions for forming the dabsyl derivatives are: reagent concentration, 1.75.10(-3) M; pH, 8.2; temperature, 70 degrees C; h eating time (t(h)), 21 min. Under these conditions good chromatographic rep eatability is obtained.