Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies

Fast Blue B Salt is a multicharged basic dye with a variety of applications in chromatographic testing, including in thin layer chromatography and lipase activity testing. 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.

High Performance Liquid Chromatography (HPLC) Method for Analysis of DAPI

Condition

Column	BIST™ B+, 4.6×150 mm, 5µm, 100A
Mobile Phase	Gradient MeCN
Buffer	H2SO4 – 0.2%
Flow Rate	1.0 ml/min
Detection	UV 320 nm
Retention time	6.01 min

Description

Class of Compounds	Dye
Analyzing Compounds	Fast Blue B Salt