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HPLC Analysis of Phenolic Compounds and Flavonoids with Overlapping Peaks

Luke Mizzi1*orcid tiny, Christina Chatzitzika1orcid tiny, Ruben Gatt2orcid tinyand Vasilis Valdramidis1orcid tiny

1Department of Food Studies and Environmental Health, Faculty of Health Sciences, University of Malta, Triq il-Qroqq, Msida MSD 2080, Malta

2Metamaterials Unit, Faculty of Science, University of Malta, Triq il-Qroqq, Msida MSD 2080, Malta

Article history:

Received: 2 June 2019

Accepted: 17 February 2020

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Key words:

HPLC analysis, UV-Vis absorbance, quantification of phenolic compounds and flavonoids, overlapping peaks


The identification and quantification of phenolic compounds and flavonoids in variousnatural food products is typically conducted using HPLC analysis. Their analysis is particularlycomplex since most natural food products contain a large number of different phenoliccompounds, many of which have similar chemical characteristics such as polarity,which makes complete separation of all eluents extremely difficult. In this work we presentand validate a method for the quantitative determination of the concentration of twocompounds with similar retention times, i.e. they show overlapping peaks in a mixed solution.Two pairs of phenolic compounds were investigated: caffeic and vanillic acids and ferulicand p-coumaric acids. This technique takes advantage of the different absorbancesof the two phenolic compounds in the eluent at various wavelengths and can be used forthe quantitative determination of the concentration of these compounds even if they arenot separated in the HPLC column. The presented method could be used to interpret theresults of HPLC analysis of food products which possess a vast spectrum of phenolic compoundsand flavonoids.

*Corresponding author:

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