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Analysis of phenolic compounds in different parts of pomegranate (Punica granatum) fruit by HPLC-PDA-ESI/MS and evaluation of their antioxidant activity: application to different Italian varieties

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Abstract

The analysis of pomegranate phenolic compounds belonging to different classes in different fruit parts was performed by high-performance liquid chromatography coupled with photodiode array and mass spectrometry detection. Two different separation methods were optimized for the analysis of anthocyanins and hydrolyzable tannins along with phenolic acids and flavonoids. Two C18 columns, core–shell and fully porous particle stationary phases, were used. The parameters for separation of phenolic compounds were optimized considering chromatographic resolution and analysis time. Thirty-five phenolic compounds were found, and 28 of them were tentatively identified as belonging to four different phenolic compound classes; namely, anthocyanins, phenolic acids, hydrolyzable tannins, and flavonoids. Quantitative analysis was performed with a mixture of nine phenolic compounds belonging to phenolic compound classes representative of pomegranate. The method was then fully validated in terms of retention time precision, expressed as the relative standard deviation, limit of detection, limit of quantification, and linearity range. Phenolic compounds were analyzed directly in pomegranate juice, and after solvent extraction with a mixture of water and methanol with a small percentage of acid in peel and pulp samples. The accuracy of the extraction method was also assessed, and satisfactory values were obtained. Finally, the method was used to study identified analytes in pomegranate juice, peel, and pulp of six different Italian varieties and one international variety. Differences in phenolic compound profiles among the different pomegranate parts were observed. Pomegranate peel samples showed a high concentration of phenolic compounds, ellagitannins being the most abundant ones, with respect to pulp and juice samples for each variety. With the same samples, total phenols and antioxidant activity were evaluated through colorimetric assays, and the results were correlated among them.

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Acknowledgements

The authors thank Shimadzu for constantly supporting their research work. The authors also thank Luca Santi for his logistic support and Maurizio Zecchini for in-field management of plants and fruits.

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Authors and Affiliations

  1. Department of Medicine, University Campus Bio-Medico of Rome, Via Álvaro del Portillo 21, 00128, Rome, Italy

    Marina Russo, Chiara Fanali, Giusy Tripodo, Paola Dugo, Laura Dugo, Laura De Gara & Luigi Mondello

  2. Chemistry, Biological, Pharmaceutical and Environmental Sciences Department, University of Messina, Polo Annunziata, Viale Annunziata, 98168, Messina, Italy

    Paola Dugo & Luigi Mondello

  3. Chromaleont s.r.l., Viale Boccetta 70, 98122, Messina, Italy

    Paola Dugo & Luigi Mondello

  4. Department of Agricultural and Forestry Sciences, University of Tuscia, Via S.C. DeLellis snc, 01100, Viterbo, Italy

    Rosario Muleo

  5. Tree and Timber Institute, National Research Council of Italy, Via Madonna del Piano 10, 50019, Sesto Fiorentino, Italy

    Rosario Muleo

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  1. Marina Russo
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  2. Chiara Fanali
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  6. Laura Dugo
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  7. Laura De Gara
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  8. Luigi Mondello
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Correspondence to Chiara Fanali.

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Published in the topical collection Discovery of Bioactive Compounds with guest editors Aldo Laganà, Anna Laura Capriotti and Chiara Cavaliere.

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Russo, M., Fanali, C., Tripodo, G. et al. Analysis of phenolic compounds in different parts of pomegranate (Punica granatum) fruit by HPLC-PDA-ESI/MS and evaluation of their antioxidant activity: application to different Italian varieties. Anal Bioanal Chem 410, 3507–3520 (2018). https://doi.org/10.1007/s00216-018-0854-8

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  • DOI: https://doi.org/10.1007/s00216-018-0854-8

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