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TBA Test


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5.6.6. Thiobarbituric acid (TBA)


This is one of the most widely used tests for determining the extent of lipid oxidation. It measures the concentration of relatively polar secondary reaction products, i.e., aldehydes. The lipid to be analyzed is dissolved in a suitable non-polar solvent which is contained within a flask. An aqueous solution of TBA reagent is added to the flask and the sample is shaken, which causes the polar secondary products to be dissolved in it. After shaking the aqueous phase is separated from the non-polar solvent, placed in a test-tube, and heated for 20 minutes in boiling water, which produces a pink color. The intensity of this pink color is directly related to the concentration of TBA-reactive substances in the original sample, and is determined by measuring its absorbance at 540 nm using a UV-visible spectrophotometer. The principle source of color is the formation of a complex between TBA and malonaldehyde, although some other secondary reaction products can also react with the TBA reagent. For this reason, this test is now usually referred to as the thiobarbituric acid reactive substances (TBARS) method. TBARS is an example of a measurement of the increase in concentration of secondary reaction products.




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