HPLC Method for Analysis of Allura Red (FD&C Red 40, E129) on a BIST A Column

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

HPLC Method for Analysis of Allura red on BIST A Column by SIELC Technologies
High Performance Liquid Chromatography (HPLC) Method for Sunset Yellow

Allura Red (FD&C Red 40,  E129) is a popular red synthetic dye used in a wide variety of food products. Using SIELC’s newly introduced BIST™ method, Allura Red 1, which ionizes in water, can be retained 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 Acid Red 1 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. Using this new and unique analysis method, Allura Red can be retained and UV detected at 500 nm.


ColumnBIST™ A, 4.6×150 mm, 5 µm, 100A
Mobile PhaseMeCN – 70/30%
BufferTMDAP formate pH 4.0 – 5.0 mM
Flow Rate1.0 ml/min
Detection500 nm


Class of CompoundsAcid, Dye
Analyzing CompoundsAllura red