Why crushing is done in TCA lipid peroxidation test?

In the thiobarbituric acid reactive substances (TBARS) assay for lipid peroxidation, crushing of the tissue sample is performed for several reasons:

To facilitate homogenization: Crushing the tissue sample helps in breaking down the cellular structures and releasing the lipids into the surrounding medium. This homogenization process ensures that the lipids are evenly distributed and accessible for reaction with thiobarbituric acid (TBA).

To enhance lipid extraction: Crushing the tissue sample increases the surface area of the lipids, allowing for better extraction into the organic solvent (usually a mixture of chloroform and methanol) used in the TBARS assay. This step is crucial to isolate the lipids from other cellular components and facilitate their reaction with TBA.

To release lipid peroxidation products: Crushing the tissue sample can help release lipid peroxidation products, such as malondialdehyde (MDA), which are often bound to cellular proteins or other macromolecules. By disrupting these bonds, crushing allows for the release and subsequent quantification of MDA and other TBARS.

By crushing the tissue sample, the homogenization, lipid extraction, and release of lipid peroxidation products are optimized, leading to more accurate and reliable measurements of lipid peroxidation in the TBARS assay.