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)

Application Column


BIST™ columns offer a unique and effective way to achieve separations that were traditionally challenging or even impossible with other HPLC columns. With the use of a special mobile phase, these ion exchange columns provide very strong retention for analytes with the same charge polarity as the stationary phase, unlocking new chromatography applications. What makes BIST™ columns stand out is their proprietary surface chemistry, which results in superior selectivity, resolution, and sensitivity. These columns offer a simple, efficient solution for a variety of analytical challenges, making them an excellent choice for researchers and analysts across many different fields. To learn more about the technology that powers BIST™ columns and to explore related applications, check out https://BIST.LC.

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Application Analytes:
Allura Red (Red #40)
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.