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Öğe Separation, optimization, and quantification of cytokinins by a recently developed amide-embedded stationary phase(Taylor & Francis, 2017-07-06) Aral, Hayriye; Haşimi, Duygu; Aral, Tarık; Levent, Abdulkadir; Ziyadanoğulları, BerrinIn this study, some plant growth regulators known as cytokinins [kinetin (K), zeatin (Z), thidiazuron (TDZ), benzylaminopurine (BAP), and dimethylallylaminopurine (AAP)] were separated by HPLC using an amide-embedded mixed-mode stationary phase which was synthesized by Aral et al. in recent years. The effect of mobile phase content, mobile phase pH, buffer concentration, and temperature on separation process was studied. In addition, a quantitative determination of cytokinins from Salvia limbata extract was studied, and some validation parameters such as limit of detection (LOD), limit of quantification (LOQ), and relative standard deviation (RSD) were calculated as a range of 0.03–0.1, 0.1–0.26 mg/L, and 0.03–0.08, respectively.Öğe Development of a novel amide-silica stationary phase for the reversed-phase HPLC separation of different classes of phytohormones(Elsevier, 2013-11) Aral, Hayriye; Aral, Tarık; Ziyadanoğulları, Berrin; Ziyadanoğulları, RecepA 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.
