TBARS/MDA Universal Colorimetric Detection Kit

Assay Principle: 

The TBARS/MDA Universal Colorimetric Detection Kit quantitatively measures lipid peroxidation levels in serum, plasma, tissue, cell extracts, food extracts, urine, and buffers. This detection kit has a run time of 60 minutes. Please read the complete kit insert for more information before performing this assay. Use our provided MDA Standard to generate a standard curve.

Protocol Summary:

• Introduce standards or diluted samples into the transparent microtiter plate provided.
• Add TBA Substrate to each well, ensuring thorough mixing.
• Incubate the mixture covered at 37°C with shaking for 60 minutes. This incubation facilitates the color-generating reaction between the TBA substrate and MDA in the samples.
• After the incubation period, detect the generated signal at 535nm using a plate reader. Calculate the MDA concentration using the intensity and the established standard curve.

Background:

Lipid peroxidation is a key indicator of oxidative stress in both plant and animal cells, leading to cellular injury. It involves the degradation of polyunsaturated fatty acids, resulting in several compounds, including stable malondialdehyde (MDA).

Researchers measure MDA by the reaction of thiobarbituric acid (TBA) in an acid environment according to the following reaction.

The MDA-TBA adduct formed in this reaction has a pink color and can be read at ƛ= 532 nm. Measurement between 530–545 nm is acceptable.

Beyond MDA, lipid peroxidation can also produce reactive aldehydes like 2-alkenals and 2,4-alkedienals, which react with TBA to form color. The collective measure of MDA and these substances is known as TBARS (Thiobarbituric Acid Reactive Substances). Despite some controversies regarding its specificity due to TBA’s reactivity with various aldehydes, the TBARS assay is widely used to assess lipid peroxidation. The degree of lipid unsaturation in samples typically correlates with the TBARS values, making this assay particularly useful in diverse oxidative stress studies. In general, lipids with greater unsaturation will yield higher TBARS values.

The DetectX® TBARS/MDA Universal Colorimetric Detection Kit is a useful tool for researchers studying oxidative stress, lipid metabolism, and related cellular damage, offering a reliable method for quantifying lipid peroxidation in a wide range of sample types.