HPLC Method for Chicago Sky Blue 6B on BIST A+ by SIELC Technologies

Chicago Skye Blue 6B is an organic dye also known as Direct Blue 1 and has the chemical formula C₃₄H₂₄N₆Na₄O₁₆S₄. It is typically sold as a blue powder and is primarily used in histochemistry. Commercially, it is also used in dyeing textiles with high contents of cellulose. You can find detailed UV spectra of Chicago Skye Blue 6B and information about its various lambda maxima by visiting the following link.

Chicago Sky Blue 6B can be retained and separated into its component compounds easily on a negatively-charged, cation-exchange BIST A+ column. There are two keys to this retention method: 1) a multi-charged, positive buffer, such as N,N,N’,N’-Tetramethyl-1,3-propanediamine (TMDAP), which acts as a bridge, linking the negatively-charged anion analytes to the negatively-charged column surface and 2) a mobile phase consisting mostly of organic solvent (such as MeCN) to minimize the formation of a solvation layer around the charged analytes. While both of these sulfonated dyes have the same charge, their molecular structure (and therefore charge distribution) differs just enough to allow them to be separated in a high-organic MP. 

Other positively-charged buffers that can generate BIST™ include DMP, Calcium acetate, and Magnesium acetate. Using this new and unique analysis method, Brilliant Black BN and Chicago Sky Blue can be retained and separated with high selectivity and great peak shape. This method can be detected and is compatible with ELSD, CAD, and Mass Spectrometry (LC-MS).

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