title: The pentafluorophenyl stationary phase shows a unique separation efficiency for performing fast chromatography determination of highbush blueberry anthocyanins.
abstract: A high-performance liquid chromatography method using an alternative pentafluorophenyl (PFP) core-shell stationary phase has been developed and used for rapid separation of 23 anthocyanins in a highbush blueberry Bluehaven cultivar. A high efficiency of separation of anthocyanins was achieved in the core-shell column Kinetex PFP, 150×4.6mm (particle size 2.6µm) with a 5×4.6mm precolumn, using a simple linear gradient elution with a mobile phase of acetonitrile and a water solution of 2% formic acid at a flow rate of 1.0ml/min and at a temperature of 50°C. The detection wavelength was set at 520nm for detection of all anthocyanins. The homogenized blueberry sample (Bluehaven cultivar) was extracted using pure methanol with 1.3% formic acid using an ultrasound bath for 20min and then filtrated. A 5-µL sample volume was directly injected into the HPLC system. The developed method showed an efficient separation of 23 anthocyanins in a total runtime of 21min. The potential of the pentafluorophenyl phase for efficient separation was demonstrated on a wide range of anthocyanins varying in glycosylation and acylation patterns found in highbush blueberries. The fluorinated stationary phase showed an alternative and complementary separation approach providing unique aromatic and polar selectivity in comparison with common C-18 phases.
authors.affiliation: The Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Heyrovského 1203, Hradec Králové 500 05, Czech Republic; The Research and Breeding Institute of Pomology Holovousy, Holovousy 129, Hořice 508 01, Czech Republic.
authors.forename: Barbora
authors.initials: B
authors.lastname: Šmídová
authors.affiliation: The Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Heyrovského 1203, Hradec Králové 500 05, Czech Republic. Electronic address: satinsky@faf.cuni.cz.
authors.forename: Dalibor
authors.initials: D
authors.lastname: Šatínský
authors.affiliation: The Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Heyrovského 1203, Hradec Králové 500 05, Czech Republic.
authors.forename: Kateřina
authors.initials: K
authors.lastname: Dostálová
authors.affiliation: The Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Heyrovského 1203, Hradec Králové 500 05, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Solich
mesh_terms: D000872:Anthocyanins / Q000032:analysis* / Q000302:isolation & purification*; D029798:Blueberry Plants / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D054879:Halogenation; D013997:Time Factors
chemical_list: D000872:Anthocyanins
keywords: Anthocyanins; Blueberry; Chromatography; Core-shell columns; Pentafluorophenyl (PFP); Separation