Branka Mozetič and Polonca Trebše
Nova Gorica Polytechnic, Laboratory for Environmental Research,
P.Box. 301, Nova Gorica 5001-SI,
In absence of standards, HPLC coupled with DAD offers identification of polyphenols by scanning UV-Vis spectra of individual components, which spectral characteristics are unique, but not selective. At the same time HPLC-DAD determination methods of polyphenols differ in mobile phase solutions resulting in DAD scanned spectra deviation between different studies, aggravating the precise identification based on agreement to UV-Vis data from literature. Mass spectrometry (MS) detection with molar weight determination of the individual components in the sample enables more precise identification of compounds eluted from the column. Sweet cherry Petrovka polyphenols were separated on C18 Hypersil PEP 300 column (250 x 4.6 mm, 5μm) using gradient solvent mixture consisting of methanol, water and formic acid. MS and UV-Vis spectra of eluted anthocyanins and hydroxycinnamic acids were recorded. HPLC-MS analyses were performed using a LCQTM ion trap, Finnigan, MAT mass spectrometer by atmospheric pressure chemical ionisation (ACPI). Molecular and fragmented ion masses of sweet cherry hydroxycinnamic acids and anthocyanins were determined and with UV-Vis spectra, in the range of 190-600 nm, used for identification of compounds. Electro spray mass spectrum of two hydroxycinnamic acids produced ions with m/z ratios of 353.0 and 337.0, which corresponded to molecular weights of neochlorogenic acid and 3’-p-coumarylquinic acid. The molecular weights of 5 anthocyanins corresponded to cyanidin-3-glucoside (449.0), cyanidin-3-rutinoside (595.1), peonidin-3-glucoside (463.1), pelargonidin-3-rutinoside (579.1) and peonidin-3-rutinoside (609.2).
Key words: sweet cherries, anthocyanins, hydroxycinnamic acids, HPLC-DAD/MS