HPLC Method for Analysis of Fast Blue B Salt on BIST B+ by SIELC Technologies.
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.
Condition
| Column | BIST B+, 4.6 x 150 mm, 5 µm, 100 A, dual ended |
| Mobile Phase | Gradient MeCN |
| Buffer | H3PO4 – 0.2% |
| Flow Rate | 1.0 ml/min |
| Detection | UV 320 nm |
| Peak Retention Time | 6.01 min |
Description
| Class of Compounds | Dye |
| Analyzing Compounds | Fast Blue B Salt |
Application Column
BIST B+
Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A
Column options: dual ended
