Simultaneous analysis of indole-3-acetic acid, indole butyric acid, abscisic acid and naphthalene acetic acid by HPLC

• 1、School of Chemistry and Chemical Engineering, Henan University, Kaifeng, 475004, Henan，PR China

Abstract：A high performance liquid chromatography (HPLC) method was established for the simultaneous separation and determination of indole-3-acetic acid (IAA), indole butyric acid (IBA), abscisic acid (ABA) and naphthalene acetic acid (NAA). Using Eclipse XDB-C8（4.6 mm*250 mm, 5 μm）reversed-phase column as seperation column. The mobile phase composed of 70% methanol and 30% buffer (formic acid and ammonium formate solution, pH=3.2), the flow rate was 1 mL/min, the injection volume was 5 μL, the temperature of the column was set at 25 ℃, and the detector wavelength was 280 nm. Under optimized experimental conditons, The results showed that the separation effect of the four hormones was satisfactory, and the correlation coefficient was in the linear range for 0.9979-0.9983, The detection limit was 0.03-0.1μg/mL, the recovery rate reached 95.8 %-104.6 % of standard addition, The relative standard deviation was not higher than 0.53%. The developed method was appied to analysis real samples, 3- indole acetic acid in bean sprouts and abscisic acid in pear were 16.8 mg/kg and 5.0 mg/kg.

Keywords： Analytical Chemistry, HPLC, Plant Hormones, Analysis, Detection