Optimized High-Performance Liquid Chromatography Method for Determining Nine Cytokinins, Indole-3-acetic Acid and Abscisic Acid

Abstract

Liquid-liquid extraction and solid phase extraction followed by high-performance liquid chromatography (HPLC) connected with ultraviolet (UV) detection were used for the determination of phytohormones. The parameters influencing the performance of the HPLC-UV method, including composition of the mobile phase for gradient elution, column temperature, flow rate, and detection wavelength, were optimized. This method can simultaneously determine 11 phytohormones, including nine cytokinins, indole-3-acetic acid, and abscisic acid. The limit of detection of this method is 0.22 to 1.1 µg L−1, and the coefficient factors of linear regression are >0.998. The recoveries of the target phytohormones ranged between 62.1~109.4%, and the relative standard deviations were <10%. This method is suitable for determining phytohormones, especially cytokinins, in young panicles, roots, and xylem sap of rice plants.