title: Development of a novel amide-silica stationary phase for the reversed-phase HPLC separation of different classes of phytohormones.
abstract: A novel amide-bonded silica stationary phase was prepared starting from N-Boc-phenylalanine, cyclohexylamine and spherical silica gel (4 µm, 60 Å). The amide ligand was synthesised with high yield. The resulting amide bonded stationary phase was characterised by SEM, IR and elemental analysis. The resulting selector bearing a polar amide group is used for the reversed-phase chromatography separation of different classes of thirteen phytohormones (plant hormones). The chromatographic behaviours of these analytes on the amide-silica stationary phase were compared with those of RP-C18 column under same conditions. The effects of different separation conditions, such as mobile phase, pH value, flow rate and temperature, on the separation and retention behaviours of the 13 phytohormones in this system were studied. The optimum separation was achieved using reversed-phase HPLC gradient elution with an aqueous mobile phase containing pH=6.85 potassium phosphate buffer (20 mM) and acetonitrile with a 22 °C column temperature. Under these experimental conditions, the 12 phytohormones could be separated and detected at 230 or 270 nm within 26 min.
authors.affiliation: University of Batman, Faculty of Science and Art, Department of Chemistry, Batman, Turkey.
authors.forename: Hayriye
authors.initials: H
authors.lastname: Aral
authors.forename: Tarık
authors.initials: T
authors.lastname: Aral
authors.forename: Berrin
authors.initials: B
authors.lastname: Ziyadanoğulları
authors.forename: Recep
authors.initials: R
authors.lastname: Ziyadanoğulları
mesh_terms: D000097:Acetonitriles; D000577:Amides / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D056148:Chromatography, Reverse-Phase / Q000295:instrumentation*; D006863:Hydrogen-Ion Concentration; D010937:Plant Growth Regulators / Q000145:classification / Q000302:isolation & purification*; D012822:Silicon Dioxide / Q000737:chemistry*; D055672:Static Electricity; D013696:Temperature
chemical_list: D000097:Acetonitriles; D000577:Amides; D010937:Plant Growth Regulators; D012822:Silicon Dioxide; C032159:acetonitrile
keywords: 3-chloropropylated silica; 3-chloropropyltrimethoxysilane; Amide embedded; Amide stationary phase; Boc; CP-Si; CPTMS; DCC; DCM; EtOAc; H; HILIC; HPLC; HPLC separation; Herbicides; NPLC; ODS; Plant growth regulator; Pr; Propyl; RPLC; Reversed-phase; SP; Stationary phase; TFA; dichloromethane; dicyclohexylcarbodiimide; ethyl acetate; hexane; high-performance liquid chromatography; hydrophilic interaction chromatography; normal-phase liquid chromatography; octadecylsilyl; reversed-phase liquid chromatography; tert-butoxycarbonyl; trifluoroacetic acid