AbstractIntroductionPomegranate‐husk is the main by‐product generated from the pomegranate industry. It is a potential source of compounds highly appreciated by different costumers. Punicalagin is the main compound present in pomegranate‐husk.ObjectiveTo characterise the pomegranate‐husk total polyphenols by HPLC‐ESI‐MS and to establish a method for the recovery of punicalagin using a medium pressure liquid chromatography (MPLC) system.Materials and methodsThe characterisation of total pomegranate‐husk polyphenols was carried out using liquid chromatography coupled to mass spectrometry. Thus, 200 mg of pomegranate‐husk polyphenols were fractionated by MPLC. The isolated punicalagin was characterised by HPLC‐MS and was tested as standard reagent for the measurement of its scavenging capacity reducing DPPH and ABTS radicals.ResultsTwenty peaks were identified by analytical HPLC‐MS analysis from the pomegranate‐husk polyphenols. The main compounds were the punicalagin anomers, punicalin and ellagic acid. The MPLC method allowed three fractions to be obtained. In fraction three 39.40 ± 8.06 mg of punicalagin anomers (purity > 97.9%) were recovered. The scavenging capacity of punicalagin showed an IC50 of 109.53 and 151.50 μg/mL for DPPH and ABTS radicals, respectively.ConclusionThe MPLC system was an excellent tool for the separation of the main ellagitannins from pomegranate husk and for the isolation of punicalagin anomers. Fraction three was rich in high purity punicalagin anomers. The IC50 was obtained for DPPH and ABTS radicals. Copyright © 2017 John Wiley & Sons, Ltd.