title: Metabolite fingerprinting of Punica granatum L. (pomegranate) polyphenols by means of high-performance liquid chromatography with diode array and electrospray ionization-mass spectrometry detection.
abstract: The present study was aimed at the development of a new analytical method for the comprehensive multi-component analysis of polyphenols in Punica granatum L. (pomegranate) juice and peel. While pomegranate juice was directly analysed after simple centrifugation, different extraction techniques, including maceration, heat reflux extraction, ultrasound-assisted extraction and microwave-assisted extraction, were compared in order to obtain a high yield of the target analytes from pomegranate peel. Dynamic maceration with a mixture of water and ethanol 80:20 (v/v) with 0.1% of hydrochloric acid as the extraction solvent provided the best result in terms of recovery of pomegranate secondary metabolites. The quali- and quantitative analysis of pomegranate polyphenols was performed by high-performance liquid chromatography with diode array and electrospray ionization-mass spectrometry detection. The application of fused-core column technology allowed us to obtain an improvement of the chromatographic performance in comparison with that of conventional particulate stationary phases, thus enabling a good separation of all constituents in a shorter time and with low solvent usage. The analytical method was completely validated to show compliance with the International Conference on Harmonization of Technical Requirements for the Registration of Pharmaceuticals for Human Use guidelines and successfully applied to the characterisation of commercial and experimental pomegranate samples, thus demonstrating its efficiency as a tool for the fingerprinting of this plant material. The quantitative data collected were submitted to principal component analysis, in order to highlight the possible presence of pomegranate samples with high content of secondary metabolites. From the statistical analysis, four experimental samples showed a notable content of bioactive compounds in the peels, while commercial ones still represent the best source of healthy juice.
authors.affiliation: Department of Life Sciences, University of Modena and Reggio Emilia, Via G. Campi 103, 41125, Modena, Italy.
authors.forename: Virginia
authors.initials: V
authors.lastname: Brighenti
authors.affiliation: Department of Life Sciences, University of Modena and Reggio Emilia, Via G. Campi 103, 41125, Modena, Italy; Institute of Life Science and Technology, Hanze University of Applied Sciences, Groningen (Hanze UAS), Zernikeplein 7, 9747 AS Groningen, The Netherlands.
authors.forename: Sebastiaan Frearick
authors.initials: SF
authors.lastname: Groothuis
authors.affiliation: Department of Life Sciences, University of Modena and Reggio Emilia, Via G. Campi 103, 41125, Modena, Italy.
authors.forename: Francesco Pio
authors.initials: FP
authors.lastname: Prencipe
authors.affiliation: Laboratory of Plant Science, Migal Galilee Technology Center, Southern Industrial Zone, Tarshish st. Kiryat Shmona, P.O.B. 831, Kiryat Shmona 11016, Israel; Tel-Hai College, Upper Galilee, Israel.
authors.forename: Rachel
authors.initials: R
authors.lastname: Amir
authors.affiliation: Department of Life Sciences, University of Modena and Reggio Emilia, Via G. Campi 103, 41125, Modena, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Benvenuti
authors.affiliation: Department of Life Sciences, University of Modena and Reggio Emilia, Via G. Campi 103, 41125, Modena, Italy. Electronic address: federica.pellati@unimore.it.
authors.forename: Federica
authors.initials: F
authors.lastname: Pellati
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D000067030:Fruit and Vegetable Juices / Q000032:analysis; D029561:Lythraceae / Q000737:chemistry* / Q000378:metabolism*; D059808:Polyphenols / Q000032:analysis* / Q000737:chemistry; D064210:Secondary Metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D059808:Polyphenols
keywords: HPLC; MS; Metabolite fingerprinting; Polyphenols; Pomegranate; Punica granatum L.