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A Simple HPLC/DAD Method Validation for the Quantification of Malondialdehyde in Rodent’s Brain
Authors:George Jîtc&#x;  Erzsbet Fogarasi  Bianca-Eugenia &#x;sz  Camil Eugen Vari  Amelia Tero-Vescan  Amalia Miklos  M&#x;d&#x;lina-Georgiana B&#x;trînu  Carmen Maria Rusz  Mircea Dumitru Croitoru  Maria Titica Dogaru
Abstract: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 μm) SUPELCO Column 3.3 cm × 4.6 mm and Supelco Column Saver 0.5 μm 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 μL 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 μg/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.
Keywords:oxidative stress  malondialdehyde  brain  liquid chromatography  diode array detector
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