Optimization and method validation of determining polyphenolic compounds by UFLC-DAD system using two biphenyl and pentafluorophenylpropyl columns

Abstract

The development of efficient analytical methods for the identification and quantification of polyphenols in natural products is needed. The present study was aimed to optimize the polyphenols by UFLC using two different Ascentis express columns. Chromatographic separation was performed using UFLC-DAD connected with Ascentis Express Biphenyl column (system I) and Ascentis Express F5 pentafluorophenylpropyl (PFP) column (system II). Multistep gradient elution program was used in both analytical instruments to detect 34 compounds of different classes (phenolic acids, flavonoids, isoflavones, flavanols, flavonols and flavanones). The detection wavelength, linear calibration range, theoretical plate, tailing factor, limits of detection and of quantification were determined. The results were validated and the optimized method was proven to be precise, revealing good separation in the UFLC system II using Ascentis Express F5 PFP column. In conclusion, this study shows analytical preciseness and is useful in determining polyphenols in natural products.