title: HPLC-HRMS method for fast phytochelatins determination in plants. Application to analysis of Clinopodium vulgare L.
abstract: An optimized analytical method based on C8 core-shell reverse phase chromatographic separation and high resolution mass spectral (HRMS) detection is developed for a fast analysis of unbound phytochelatins (PCs) in plants. Its application to analysis of Clinopodium vulgare L. is demonstrated where proper PCs liberating and preservation conditions were employed using dithiotreitol in the extraction step. A baseline separation of glutathione (GSH) and phytochelatins from 2 to 5 (PC2-PC5) for 3 min was achieved at conventional HPLC backpressure, with detection limits from 3 ppt (for GSH) to 2.5 ppb (for PC5). It is shown, that the use of HRMS with tandem mass spectral (MS/MS) capabilities permits additional wide range screening ability for iso-phytochelatins and PC similar compounds, based on exact mass and fragment spectra in a post acquisition manner.
authors.affiliation: Sofia University "Kliment Ohridski", Faculty of Chemistry and Pharmacy, Chair of Analytical Chemistry, 1, James Bourchier Boulevard, 1164 Sofia, Bulgaria. Electronic address: krum.bardarov@chem.uni-sofia.bg.
authors.forename: Krum
authors.initials: K
authors.lastname: Bardarov
authors.affiliation: University of Plovdiv, Faculty of Biology, Department of Plant Physiology and Molecular Biology, 24 Tsar Asen Street, 4000 Plovdiv, Bulgaria.
authors.forename: Mladen
authors.initials: M
authors.lastname: Naydenov
authors.affiliation: Sofia University "Kliment Ohridski", Faculty of Chemistry and Pharmacy, Chair of Analytical Chemistry, 1, James Bourchier Boulevard, 1164 Sofia, Bulgaria.
authors.forename: Rumyana
authors.initials: R
authors.lastname: Djingova
mesh_terms: D002851:Chromatography, High Pressure Liquid; D019686:Lamiaceae; D013058:Mass Spectrometry / Q000379:methods; D054811:Phytochelatins / Q000302:isolation & purification*; D010936:Plant Extracts / Q000737:chemistry; D018515:Plant Leaves / Q000737:chemistry*; D018517:Plant Roots / Q000737:chemistry*; D018520:Plant Shoots / Q000737:chemistry*
chemical_list: D010936:Plant Extracts; D054811:Phytochelatins
keywords: Cd, Cu, Zn and Pb stress in plants; HR–MS/MS; Iso-phytochelatins screening; Phytochelatins determination