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Two vials containing the A- and β-chains of human insulin without labels. If there is no...

Two vials containing the A- and β-chains of human insulin without labels. If there is no reference standard, Can I use TLC or UV spectroscopy to distinguish/determine A- and β-chains of human insulin? why or why not? If I can, what will be the results?

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Answer #1

Insulin is the eminent polpeptide drug. In the pancreas, the polypeptide which is internally composed has alpha and beta chains. These two chains are connected by two intersulphide bonds, and one intra sulphide bond in the alpha chain.

The label free detection of human chains of insulin can be carried out with UV spectroscopy. When the human insulin is subjected to constant exposure of UV light, the excitation of human insulin gives the datas regarding the structure, process, fluorescence properties and its absorption. In the original human insulin Alpha and Beta chains are connected by a disulphide bridge. Upon UV excitation these bridge gets broken and the structure of insulin gets dispruted. After the homolytic cleavage, mass of the two chains should be measured. From the two the chain with lower molecular mass is the Beta chain.

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