Chromatography-Free Ultrasensitive Profiling of Multiclass Plant Hormones via One-Pot Dual-Channel Derivatization...

Plant hormones regulate diverse physiological processes, but their simultaneous analysis is challenging due to structural diversity, low abundance, and complex matrices. Herein, we report a chromatography-free analytical platform integrating one-pot dual-channel derivatization with acoustic ejection mass spectrometry (AEMS). Using N,N-diethyl-1,2-ethanediamine for carboxyl-containing hormones and 2-methyl-4-phenylaminomethyl-benzeneboronic acid for cis-diol-containing brassinosteroids, 11 key phytohormones were rapidly derivatized (< 1 min) in a single reaction vessel. The derivatization enhanced sensitivity up to 183-fold, achieving femtogram-level limits of detection (3.9-22 fg). Coupled with AEMS, each sample was analyzed in ∼3 s without chromatography. Validation confirmed excellent linearity (R² ≥ 0.9991), precision (RSD < 15.3%), accuracy (77.3-110.8% recovery), and minimal matrix effects (76.3-105.1%). The method successfully quantified endogenous plant hormones in Arabidopsis thaliana seeds, Brassica napus stamens, and Oryza sativa seeds, and captured dynamic hormone changes during rice germination. This high-throughput platform provides a promising tool for comprehensive plant hormone profiling.