title: Development of a new high-performance liquid chromatography method with diode array and electrospray ionization-mass spectrometry detection for the metabolite fingerprinting of bioactive compounds in Humulus lupulus L.
abstract: The study was aimed at developing a new analytical method for the metabolite fingerprinting of bioactive compounds in Humulus lupulus L. (hop), together with a simple extraction procedure. Different extraction techniques, including maceration, heat reflux extraction (HRE), ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE), were compared in order to obtain a high yield of the target analytes. Dynamic maceration for 30min with MeOH-HCOOH (99:1, v/v) as the extraction solvent provided the best result in terms of recovery of secondary metabolites. The analysis of hop constituents, including prenylflavonoids and prenylphloroglucinols (bitter acids), was carried out by means of HPLC-UV/DAD, HPLC-ESI-MS and MS(2), using an ion trap mass analyzer. An Ascentis Express C18 column (150mm×3.0mm I.D., 2.7μm) was used for the HPLC analysis, with a mobile phase composed of 0.25% formic acid in both water and acetonitrile, under gradient elution. The method validation was performed to show compliance with ICH guidelines. The validated technique was successfully applied to the phytochemical analysis of ten commercial cultivars and twenty-three wild Italian hop genotypes, thus demonstrating to be a reliable and useful tool for the comprehensive multi-component analysis of hop secondary metabolites.
authors.affiliation: Department of Life Sciences, University of Modena and Reggio Emilia, Via G. Campi 183, 41125 Modena, Italy.
authors.forename: Francesco Pio
authors.initials: FP
authors.lastname: Prencipe
authors.affiliation: Department of Life Sciences, University of Modena and Reggio Emilia, Via G. Campi 183, 41125 Modena, Italy.
authors.forename: Virginia
authors.initials: V
authors.lastname: Brighenti
authors.affiliation: Department of Food Science-LS9 Interlab Group, University of Parma, Via G.B. Usberti 95/a, 43134 Parma, Italy.
authors.forename: Margherita
authors.initials: M
authors.lastname: Rodolfi
authors.affiliation: Department of Food Science-LS9 Interlab Group, University of Parma, Via G.B. Usberti 95/a, 43134 Parma, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Mongelli
authors.affiliation: Department of Food Science-LS9 Interlab Group, University of Parma, Via G.B. Usberti 95/a, 43134 Parma, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: dall'Asta
authors.affiliation: Department of Food Science-LS9 Interlab Group, University of Parma, Via G.B. Usberti 95/a, 43134 Parma, Italy.
authors.forename: Tommaso
authors.initials: T
authors.lastname: Ganino
authors.affiliation: Department of Food Science-LS9 Interlab Group, University of Parma, Via G.B. Usberti 95/a, 43134 Parma, Italy.
authors.forename: Renato
authors.initials: R
authors.lastname: Bruni
authors.affiliation: Department of Life Sciences, University of Modena and Reggio Emilia, Via G. Campi 183, 41125 Modena, Italy. Electronic address: federica.pellati@unimore.it.
authors.forename: Federica
authors.initials: F
authors.lastname: Pellati
mesh_terms: D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D005419:Flavonoids / Q000032:analysis; D027582:Humulus / Q000737:chemistry*; D010696:Phloroglucinol / Q000032:analysis; D010936:Plant Extracts / Q000737:chemistry*; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D005419:Flavonoids; D010936:Plant Extracts; D010696:Phloroglucinol
keywords: Bitter acids; Fused-core; HPLC; Hops; Humulus lupulus; Prenylflavonoids