A Simple HPLC/DAD Method Validation for the Quantification of Malondialdehyde in Rodent's Brain

In the present study, a HPLC/DAD method was set up to allow for the determination and quantification of malondialdehyde (MDA) in the brain of rodents (rats). Chromatographic separation was achieved on Supelcosil LC-18 (3 mu m) SUPELCO Column 3.3 cm x 4.6 mm and Supelco Column Saver 0.5 mu m filter by using a mobile phase acetonitrile (A) and phosphate buffer (20 mM, pH = 6) (B). Isocratic elution was 14% for (A) and 86% for (B). The injection volume (loop mode) was 100 mu L with an analysis time of 1.5 min. Flow rate was set at 1 mL/min. The eluted compound was detected at 532 nm by a DAD detector by keeping the column oven at room temperature. The results indicated that the method has good linearity in the range of 0.2-20 mu g/g. Both intra- and inter-day precision, expressed as RSD, were <= 15% and the accuracies ranged between +/- 15%. The lower limit of quantification (LLOQ), stability, and robustness were evaluated and satisfied the validation criteria. The method was successfully applied in a study of chronic toxicology following different treatment regimens with haloperidol and metformin.

Automated summary

A HPLC/DAD method was developed for the determination and quantification of MDA in the brain of rodents, showing good linearity, precision, and accuracy. The method was successfully applied in a study of chronic toxicology following different treatment regimens, demonstrating its reliability and effectiveness.