Background

Lipid peroxidation, a well-established mechanism of cellular injury in plants and animals, is used as an indicator of oxidative stress in cells and tissues. Lipid peroxides are unstable and decompose to form a complex series of compounds including reactive carbonyl compounds. Polyunsaturated fatty acid peroxides generate malondialdehyde (MDA) and 4-hydroxyalkenals (HAE) upon decomposition, and the measurement of MDA and HAE has been used as an indicator of lipid peroxidation.

Assay Principle

The method in this assay is designed to assay either MDA alone (in hydrochloric acid) or MDA in combination with HAE (in methanesulfonic acid.) The assay is based on the reaction of a chromogenic reagent, N-methyl-2-phenylindole (R1), with MDA and 4-hydroxyalkenals at 45°C. One molecule of either MDA or 4-hydroxyalkenal reacts with 2 molecules of reagent R1 to yield a stable chromophore with maximal absorbance at 586 nm.

OBR provides two methods of LPO assessment to help meet your needs. Product number FR12 represents our traditional cuvette based method accommodating 94 singlet samples while FR22 is a 96-well microplate method which accomodates 23 samples in triplicate as well as a standard curve. Each method is capable of analysis for 4-HNE or MDA in a wide array of biological samples, including serum and tissue extracts.

References

1. Esterbauer and Cheesman, Meth. Enzymol. 186: 407-421, 1990.

2. Janero, et al., Free Rad. Biol. Med. 9: 515-540, 1990