Evaluation of a sensitive HPLC method for the determination of Malondialdehyde, and application of the method to different biological materials

Abstract

Reactive oxygen species (ROS), important mediators of cell and tissue injury during inflammation, are produced by several types of inflammatory cells. The formation of ROS can be monitored by detection of lipid peroxidation products. The extremely broad spectrum of biological effects of aldehydic lipid peroxidation products has necessitated the development of a technique that enables the sensitive routine quantitation of aldehydes formed in biological materials. MalondiaJdehyde (MDA) is a by-product of enzymatic eicosanoid formation and an endproduct of nonenzymatic peroxidation of polyunsaturated fatty acids with three or more bisallylic double bonds, The determination of the thiobarbituric acid derivative of MDA (TBA-MDA) is a widely used method for estimating overall lipid peroxidation. We describe a rapid, isocratic, simple, and sensitive high-performance liquid chromatographic (HPLC) method with spectrafluorimetric detection for measurement oF MDA-TBA in human biological samples such as plasma, urine, wound secretions, amniotic fluid, sputum and tissue samples. By use of this method, picomole quantities of MDA can be readily and specifically detected in different biological materials. Coefficients of variation of repeated MDA-TBA assays were 4.4% within run and 6.9% from run to run. Reference values are given for a variety of human body fluids and for rat tissues.

Carbon tetrachloride-induced hepatic and renal damages in rat: inhibitory effects of cacao polyphenol

“…15) Briefly, sodium dodecyl sulfate, TBA, and BHT were added to the liver and kidney homogenates and mixed. After the mixture was heated in boiling water, TBA-MDA adducts were extracted with a butanol/pyridine solution and the extract was subjected to LC-FL using a COS-MOSIL 5C18-MS-II column (4.6 × 250 mm; Nacalai Tesque, Kyoto, Japan).…”

Protective effect of melatonin against multistress condition induced lipid peroxidation via measurement of gastric mucosal lesion and plasma malondialdehyde levels in rats

“…On breakdown of such hydroperoxides a great variety of aldehydes can be formed (as secondary products). MDA, a three carbon compound formed by scission of peroxidized polyunsaturated fatty acids, is one of the main secondary products of lipid peroxidation [16,22] . MDA is reactive toward amino groups of protein and nucleic acid and have cytotoxic and mutagenic effects.…”

“…The aliquots were kept frozen at -80℃ until analysis. Plasma samples in this temperature are stable for at least one year [16] . Then the stomachs were immediately removed and cut along the greater curvature and the stomach contents were washed out with saline and gastric lesions were fixed with 20% formaldehyde for 5 min.…”

In Stored Human Blood, The Inhibitor Effect of Tannic Acid and Caffeic Acid on Lipid Peroxidation and Oxidative DNA Damage

“…Six milliliters of blood was taken from each bag and four milliliters centrifuged (1100 rpm) and the plasma was obteined to measure plasma malondialdehyde (MDA), ubiquinone-10 (CoQ10) levels and two milliliters of whole blood was separated for DNA 8-hydroxy-2 deoxyguanosine (8-OHdG) measurement at the beginning (day 0) and on days 7, 14, 21, 28 by high pressure liquid chromatography (HPLC). Plasma MDA measurements by High Pressure Liquid Chromatography (HPLC): MDA measurement in plasma was performed according to the method of Khoschsor et al (14). Plasma MDA measurements by High Pressure Liquid Chromatography (HPLC): A total of 0.44 mol/L H 3 PO 4 and 42 mmol/L thiobarbituric acid (TBA) was added to 50 μL plasma and incubated for 60 minutes in boiling water.…”