Laboratory of Growth Regulators
Palacký University Olomouc Head of Research Program: Aleš Pěnčík
Auxins in Plants
The group concentrates on the development and practical application of analytical protocols suitable for study of auxins as well as their precursors and metabolites. The outstanding features of high-throughput ultra high-performance liquid chromatography (UHPLC) coupled with selective tandem mass detection (MS/MS), reaching typically 1 fmol in sensitivity, are exploited to quantify studied compounds. Preceeding the methods used to purify analytes from raw plant extracts are solid-phase extraction (SPE, both conventional and its miniaturized “in-tip” version) and/or immunoaffinity extraction (based on polyclonal as well as monoclonal antibodies). To track recovery of the developed protocols, a wide spectrum of 2H and 13C-labelled standards is used. To quantify authentic compounds isolated from plant material, the method of internal standardization is applied.
Combination of the above- methods enables analysis of auxin metabolom in various complex of plant materials. Moreover, the high specificity of immunoaffinity purification and advanced sensitivity of the instrumentation in use, allows for determination of auxin and some of its derivatives in as minute samples as tens of root tips of Arabidopsis thaliana seedlings.
Indols in Human
Interest in L-tryptophan (L-TRP) metabolites research has been increasing during the past decades because of their demonstrable effects related to human health. It was shown that many of the L-TRP metabolites, including kynurenines, indoleamines (serotonin, melatonin) and kynuramines, possess considerable neurochemical properties. Nevertheless, comprehensive data concerning the tissue and body fluids levels, as well as the dynamic state of human metabolism still remains incomplete. Our research program is focused on quantitative and qualitative ultra high-performance liquid chromatography (UHPLC) – tandem mass spectrometry (MS/MS) profiling of indolic compounds in human tissue and fluids and identification of new neuroactive substances in human. Special part of the interest is aimed at biological samples preparation by the use of efficient purification method (SPE, micro-spin centrifugation, immunoaffinity extraction). By the application of high-throughput UHPLC-MS/MS method various studies on L-TRP-metabolism and its connection with diverse neurodegenerative diseases can be accomplished.