Automated high-performance liquid chromatographic separation with spectrofluorometric detection of a malondialdehyde-thiobarbituric acid adduct in plasma.

An automated HPLC separation method for the detection of malondialdehyde (MDA) in plasma samples of a healthy population is described. The procedure involves a first step of acidic hydrolysis and complex formation with thiobarbituric acid (TBA), then a protein precipitation step, and finally the separation of TBA-MDA adducts through a complete HPLC apparatus with spectrofluorometric quantification. This procedure is very useful for a routine laboratory because of its rapidity (tR = 1.7 min), simplicity and applicability, and the method gives very good results with respect to linearity (0.1-12 mumol/l), precision (within-assay C.V.%: 8, between-assay C.V.%: 10) and accuracy (recovery average: 91.66%).