Analytical Methods for Determination of Electrophilic Intermediates

In view of the reactive properties of Electrophilic intermediates (EIs) and their involvement in various disease processes (cancer, diabetes, and drug-sensitivity), their quantitative determination is very important. This book is directed towards the development of methods for the determination of epoxides, ,-unsaturated carbonyls and ,-dicarbonyls. 1,2-Epoxy-3-butene (EB), methyl vinyl ketone (MVK), and alloxan were used as representative molecules. Following derivatization with N,N-diethyldithiocarbamate (DTC) or 1,2-phenylenediamine (PD), the stable adducts of EIs were analyzed by reversed phase (RP) high-performance liquid chromatography (HPLC) coupled with either UV or fluorescence detection. This facilitated the detection of all three compounds, viz., EB, MVK and alloxan. The detection and reproducibility were accomplished to lower picomole or femtomole levels. Further, this approach was robust and easily adaptable for the analysis of various EIs belonging to the above class of compounds. The methods were suitable for analysis of EI's in biological fluids viz., fetal bovine serum and tissue homogenates of brain, heart, liver and kidney.