Elsevier

Methods in Enzymology

Volume 233, 1994, Pages 314-318
Methods in Enzymology

[32] High-performance liquid chromatography-based thiorbarbituric acid tests

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Publisher Summary

The thiobarbituric acid (TBA) test is one of the oldest and most frequently used tests for measuring the peroxidation of fatty acids, membranes, and food products. In the food and dairy industries, the test is used as a general measure of lipid rancidity, while in medicine it remains the most commonly applied assay for lipid peroxidation to provide evidence of the involvement of free radicals in disease pathologies. The test is popular because it is simple and inexpensive. The sample under test is heated with TBA at low pH, and a pink chromogen is measured for absorbance at, or close to, 532 nm, or its fluorescence at 553 nm. The TBA test is often said to measure malondialdehyde (MDA) formed in peroxidizing lipid systems. Calibration of the test is achieved using MDA prepared in the laboratory by hydrolysis of 1,1,3,3-tetramethoxypropane or 1,1,3,3-tetraethoxypropane, and so results are frequently expressed as micromolar MDA equivalents. The availability of the technique of high-performance liquid chromatography (HPLC) is instrumental in the reevaluation of the TBA test. Many of the HPLC-based assays for the TBA test available at present have made a considerable contribution to the understanding of what the TBA test actually measures.

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