HPLC Method for Analysis of Fast Blue B Salt on BIST B+ Column

Fast Blue B Salt is a multicharged basic dye with the chemical formula C14H12Cl4N4O2Zn . It has a a variety of applications in chromatographic testing, including in thin layer chromatography, lipase activity testing. and in testing acid phosphatase in Clostridium perfringens.

Using SIELC’s newly introduced BIST™ method, Fast Blue B Salt 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 analytes to the positively-charged column surface and 2) a mobile phase consisting of a majority of organic solvent (such as MeCN) to minimize the formation of a solvation layer around the charged analytes. Utilizing a step gradient to switch to a completely aqueous MP after 2 minutes allows for retention to occur while also preventing the method from being too long. Using this new and unique analysis method, Fast Blue B Salt can be separated, retained, and UV detected at 320 nm.