The first step in determining malondialdehyde is sample preparation with derivatization. The derivatization reagent transforms malondialdehyde into a fluorescent product. Afterward, the pH is optimized through the addition of a reaction solution. 20 μl of the supernatant are injected into the HPLC system.
The separation via HPLC follows an isocratic method at 30 °C using a reversed-phase column. One run lasts 4 minutes. A fluorescence detector records the chromatograms. The quantification is performed with the delivered plasma calibrator; the concentration is calculated via the external standard method’s integration of the peak heights.