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Gas chromatographic analysis of free and bound malonaldehyde in rat liver homogenates
Author(s) -
Ichinose T.,
Miller M. G.,
Shibamoto Takayuki
Publication year - 1989
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02535765
Subject(s) - chemistry , thiobarbituric acid , chromatography , lipidology , hydrolysis , clinical chemistry , capillary gas chromatography , gas chromatography , acetic acid , lipid peroxidation , acid hydrolysis , quantitative analysis (chemistry) , biochemistry , antioxidant
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 with N ‐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 CCl 4 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 CCl 4 administration. However, the TBA values were approximately ten times the MA values in most samples.