Development of an Ultrahigh-Performance Liquid Chromatography–Triple Quadrupole Mass Spectrometry Method for Multiclass Phytohormone Quantification Via Multifunctional Chemical Derivatization

Plant growth regulation and responses to biotic and abiotic stress factors are mediated by phytohormones. Understanding the effects of the phytometabolome is essential for addressing future global agricultural and food sector challenges. Therefore, a liquid chromatography-mass spectrometry method was developed to improve the ability to determine the concentrations of various phytohormones. Using only 20 mg of plant material, a total of 27 hormones from different classes, including 18 gibberellins, abscisic acid, salicylic acid, auxin, and jasmonates, can be analyzed in 10 min via the stable isotope dilution assay approach. Using a carbodiimide-hydrazine one-pot multifunctional chemical derivatization approach, the analytes can be detected with quantitation limits ranging from 0.04 to 29.9 nM. Application of the method to tomato, maize, and cress verified its applicability, thus enabling quantitative multiclass phytohormone profiling.