Hormonal network and methodology
Speaker: Dr Ondřej Novák, Palacký University Olomouc, Czech Republic
Phytohormone metabolite profiling in plant tissues
Our work is focused on the development of high-throughput miniaturized purification methods for minute amounts of plant tissue (1-10 mg). Two novel approaches, a simple one-step purification protocol based on in-tip microSPE (micro Solid-Phase Extraction) and a class-specific miniaturized immunoaffinity chromatography method were utilized. In order to verify the effect of complex sample matrix of the whole method as well as its efficiency and analytical accuracy, the appropriate stable isotope-labelled internal standards have been prepared by the organic synthesis. We have also developed a new method based on ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) for targeted profiling of more than 100 analytes, members of the main classes of plant hormones like cytokinins, auxins, brassinosteroids, gibberellins, jasmonates and abscisates and salicylic acid. Use of our procedures can allow the quantification of plant hormones and their derivatives in very limited amounts of plant material. The new and modern analytical approaches make possible a new direction in plant hormone metabolite profiling. We believe that UHPLC-MS/MS technology can be used for fast and sensitive quantitative analysis showing reproducibility in the plant hormone profiling in different plant tissue and cell extracts.