Abstract:

A method of high performance liquid chromatography coupled with photodiode array detector (PAD) was established for the determination of the content ratio and type of 6 exogenous hormones in plant tissue culture media. The column was a μBondaPak C18 (3.9 mm×300 mm, 10 μm, Waters), the mobile phase was 140 mmol/L sodium acetate in triethyl amine buffer (pH 4.95)-acetonitrile (75:25, v/v), and the flow rate was 1.0 mL/min. The column temperature was 37 ℃, and the detection wavelength was 285 nm. The six hormones reached the baseline separation in 9 minutes. The linear relationship was very good in the range of 4-200 ng (r2>0.9995). The exogenous hormones in the medium were extracted by methanol after vacuum dried. The average recoveries of the exogenous hormones were more than 85%. The method can be used for the analysis of exogenous hormones of plant tissue culture media, or of unknown hormone ratio and the type of media.

Key words: exogenous hormone, plant tissue culture media , reversed-phase high performance liquid chromatography (RP-HPLC)