Gas chromatographic analysis of free and bound malonaldehyde in rat liver homogenates

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Abstract
A gas chromatographic method for the measurement of free and bound malonaldehyde (MA) in rat liver homogenate was developed to investigate biological damage caused by lipid peroxidation. Free MA was derivatized by reaction withN-methylhydrazine (NMH) to form 1-methylpyrazole (1-MP), which was subsequently analyzed by capillary gas chromatography (GC). Bound forms of MA were determined by first hydrolyzing samples in acetic acid, then measuring the free MA as 1-MP. Bound MA in rat liver was released within 15 min by acid hydrolysis. A slight increase in bound MA levels in rat liver was observed 1 hr after CCl4 treatment, with bound MA then returning to control levels. Free MA levels and thiobarbituric acid assay (TBA) values did not show appreciable changes over a 24-hr period after CCl4 administration. However, the TBA values were approximately ten times the MA values in most samples.