i have found two main method for sialic acid estimation.
1. Thiobarbituric acid estimation
2. Acid ninhydrin reaction method
if you have available protocol for any of the two method please provide.
Roche ® colorimetric test
It is done by means of a colorimetric test. Sialic acid is hydrolyzed by neuramidases to release free N-acetylneuraminic acid (AcNeu). In the presence of AcNeu-aldose, AcNeu is converted to N-acetylmanosamine and pyruvate, which is oxidized in the presence of MgCl2, flavin adenine dinucleotide (FAD) and thiamine pyrophosphate (TPP) by pyruvate oxidase to acetyl phosphate, CO2 and peroxide. hydrogen. The amount of hydrogen peroxide formed will be equivalent to free N-acetylneuraminic acid (AcNeu), which is converted by peroxidase (POD) in the presence of 4-aminoantipyrine (Ha-AAP) and N-ethyl-N-2-hydroxyethyl- 3-toluidine (EHTM) to a colorful compound (red), which absorbs at 550 nm.
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