HPLC Method for Analysis of Cytochrome C on BIST™ B+ Column

Cytochrome C , also known as cytochrome complex , is a mitochondrial protein that is essential to both the respiratory electron transport chain and the apoptosis cycle of cells with the chemical formula C42H52FeN8O6S2 . It is a small hemeprotein that is usually associated with the inner membrane of the mitochondrion. It is primarily used to detect peroxide production in biological systems.

Using SIELC’s newly introduced BIST™ method, this essential protein can be retained on a positively-charged anion-exchange BIST B+ column. There are two keys to this retention method: 1) a multi-charged, negative buffer, such as Sulfuric acid (H2SO4), which acts as a bridge, linking the positively-charged amine analytes to the positively-charged column surface and 2) a mobile phase consisting mostly of organic solvent to minimize the formation of a solvation layer around the charged analytes. Using this new and unique analysis method, Cytochrome C can be retained and UV detected at 395 nm.