HPLC Separation of Coumaric acids on Newcrom BH Column

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Chromatogram

Description

High Performance Liquid Chromatography (HPLC) Method for Analysis of m-Coumaric acid , o-coumaric acid , p-coumaric acid .

p -Coumaric acid is a major phenolic acid with the chemical formula C 9 H 8 O 3 . It can be found in a wide variety of graminaceous plants. p -Coumaric acid is a natural metabolite contained in many edible plants and its antioxidant activities in reducing oxidative stress and inflammatory reactions have been demonstrated in various experiments.

m -Coumaric acid is a hydroxycinnamic acid with the chemical formula C 9 H 8 O 3 . It can be found in a vinegar.  It differs from p -Coumaric acid in the position of the hydroxy substitution.

o -Coumaric acid is a hydroxycinnamic acid with the chemical formula C 9 H 8 O 3 . It can be found in a vinegar.  It differs from p -Coumaric acid and m-Coumaric acid in the position of the hydroxy substitution.

The three isomers of coumaric acid, m-Coumaric acid , o-coumaric acid , p-coumaric acid , can be detected at low UV.  Using Newcrom BH mixed-mode column and a mobile phase consisting of acetonitrile (ACN), and water with phosphoric acid (H3PO4) buffer, the acids can be retained and UV detected at 270nm.

Method Parameters

Mobile PhaseMeCN/H2O – 20/80%
BufferH3PO4 – 1.0%
Flow Rate1.0 ml/min
Detection270 nm
Class of CompoundsAcid
Analyzing Compoundsm-Coumaric acid,o-coumaric acid,p-coumaric acid

HPLC Column Used

Newcrom BH, 4.6 x 150 mm, 5 µm, 100 A, dual ended