Fluorescein-core piperazine as a fluorescent labeling reagent for highly sensitive determination of jasmonic acid by high-performance liquid chromatography

Abstract

In the present work, a pre-column fluorescent derivatization reagent, fluorescein-core piperazine (FCP) was prepared for labeling jasmonic acid (JA). Based on the enhanced fluorescence after derivatization, a simple and sensitive method was developed for determination of JA by high performance liquid chromatography with fluorescence detection (HPLC/FD). Several key derivatization conditions were examined. The results indicated that FCP labeled JA could be well separated from FCP in the sample within 13 min on a C₁₈ reversed-phase column using a mobile phase of methanol–water (55 : 45, v/v) with fluorescence detection at excitation and emission wavelength of 507 nm and 526 nm, respectively. The detection limit obtained was 0.4 nM (signal to noise ratio of 3), which was lower than most of the existing methods. The method is reproducible and yields single peak for JA regardless of isomer. The specificity of the proposed method was confirmed by high-performance liquid chromatography-ion trap mass spectrometry (HPLC/IT-MS). Finally, it has been demonstrated that the proposed HPLC/FD method can be applied to the assay of JA in plant samples.