Screening and optimization of derivatization in heating block for the determination of aliphatic aldehydes by HPLC

For the study of the derivatization behavior of aliphatic C1–C10 aldehydes with 2,4-dinitrophenylhydrazine (DNPH) in a heating block a gradient elution HPLC method for separation and determination by UV detection at 360 nm was applied. The influence of time, temperature, excess of reagent and stirring onto the reaction yields were examined for conducting the reaction in a thermostated heating block. First, the derivatization procedure had been screened by experiments according to a complete factorial design in order to evaluate the statistically significant variables. Those parameters were used to establish the optimum conditions for the reaction by means of a Box-Behnken design.