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

High Performance Liquid Chromatography (HPLC) Method for Analyses of Allura Red (FD&C Red 40, E129)

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, 5µm, 100A
Mobile PhaseMeCN
BufferTMDAP phosphate – 5 mM  pH 4.0
Flow Rate2.0 mL/min
DetectionVis 600 nm


Class of CompoundsDyes
Analyzing CompoundsAllura Red (FD&C Red 40, E129)
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.


SIELC Technologies’ BIST™ Columns are a new and simple way to achieve many separations that are traditionally difficult or impossible to achieve with any other HPLC columns currently on the market! When used with our BIST™ mobile phases, these ion exchange columns can generate very strong retention of analytes that have the same charge polarity as the stationary phase, unlocking new chromatography applications that were previously too difficult to achieve.

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Application Analytes:
Allura Red (Red #40)