HPLC Method for Analysis of Tartrazine on BIST A Column

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Chromatogram

Description

· High Performance Liquid Chromatography (HPLC) Method for Analysis of Tartrazine (FD&C Yellow 5) .

Tartrazine is the most popular yellow synthetic dye in the world with the chemical formula C 16 H 9 N 4 Na 3 O 9 S 2 . It is used in a multitude of food, cosmetic products, and medications. It is one of the oldest known members of the pyrazolone family of dyes. You can find detailed UV spectra of Tartrazine (FD&C Yellow 5) and information about its various lambda maxima by visiting the following link.

Using SIELC’s newly introduced BIST™ method, Tartrazine (FD&C Yellow 5) , 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 tartrazine 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. The effect of reducing the solvation layer by increasing the organic component concentration in the mobile phase can be clearly seen above, as can the effect of multi- and singly-charged buffers on retention. Other positively-charged buffers that can generate BIST™ include Calcium acetate and Magnesium acetate. Using this new and unique analysis method, tartrazine can be retained and UV detected at 260 nm.

Method Parameters

Mobile PhaseMeCN
BufferTMDAP formate pH 4.0 – 5,0 mM
Flow Rate1.0 ml/min
Detection260 nm
Class of CompoundsAcid, Dye
Analyzing CompoundsTartrazine (FD&C Yellow 5)

HPLC Column Used

BIST A, 4.6 x 150 mm, 5 µm, 100 A, dual ended