title: Simultaneous analysis of apolar phytohormones and 1-aminocyclopropan-1-carboxylic acid by high performance liquid chromatography/electrospray negative ion tandem mass spectrometry via 9-fluorenylmethoxycarbonyl chloride derivatization.
abstract: A strategy to detect and quantify the polar ethylene precursor 1-aminocyclopropan-1-carboxylic acid (ACC) along with the more apolar phytohormones abscisic acid (ABA), indole-3-acetic acid (IAA), jasmonic acid (JA), jasmonic acid-isoleucine conjugate (JA-Ile), 12-oxo-phytodienoic acid (OPDA), trans-zeatin, and trans-zeatin 9-riboside using a single extraction is presented. Solid phase resins commonly employed for extraction of phytohormones do not allow the recovery of ACC. We circumvent this problem by attaching an apolar group to ACC via derivatization with the amino group specific reagent 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl). Derivatization in the methanolic crude extract does not modify other phytohormones. The derivatized ACC could be purified and detected together with the more apolar phytohormones using common solid phase extraction resins and reverse phase HPLC/electrospray negative ion tandem mass spectrometry. The limit of detection was in the low nanomolar range for all phytohormones, a sensitivity sufficient to accurately determine the phytohormone levels from less than 50mg (fresh weight) of Arabidopsis thaliana and Nicotiana benthamiana tissues. Comparison with previously published phytohormone levels and the reported changes in phytohormone levels after stress treatments confirmed the accuracy of the method.
authors.affiliation: Department of Molecular Signal Processing, Leibniz-Institute of Plant Biochemistry, Weinberg 3, D-06120 Halle, Germany. Electronic address: joerg.ziegler@ipb-halle.de.
authors.forename: Jörg
authors.initials: J
authors.lastname: Ziegler
authors.affiliation: Department of Molecular Signal Processing, Leibniz-Institute of Plant Biochemistry, Weinberg 3, D-06120 Halle, Germany. Electronic address: jakob.qwegwer@ipb-halle.de.
authors.forename: Jakob
authors.initials: J
authors.lastname: Qwegwer
authors.affiliation: Department of Molecular Signal Processing, Leibniz-Institute of Plant Biochemistry, Weinberg 3, D-06120 Halle, Germany. Electronic address: melvin.schubert@ipb-halle.de.
authors.forename: Melvin
authors.initials: M
authors.lastname: Schubert
authors.affiliation: Institute of Biology - Plant Physiology, Martin-Luther University Halle-Wittenberg, Weinbergweg 10, D-06120 Halle, Germany. Electronic address: jessica.erickson2525@gmail.com.
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Erickson
authors.affiliation: Institute of Biology - Plant Physiology, Martin-Luther University Halle-Wittenberg, Weinbergweg 10, D-06120 Halle, Germany. Electronic address: martin.schattat@pflanzenphys.uni-halle.de.
authors.forename: Martin
authors.initials: M
authors.lastname: Schattat
authors.affiliation: Department of Molecular Signal Processing, Leibniz-Institute of Plant Biochemistry, Weinberg 3, D-06120 Halle, Germany. Electronic address: katharina.buerstenbinder@ipb-halle.de.
authors.forename: Katharina
authors.initials: K
authors.lastname: Bürstenbinder
authors.affiliation: Department of Molecular Signal Processing, Leibniz-Institute of Plant Biochemistry, Weinberg 3, D-06120 Halle, Germany. Electronic address: cgrubb@ipb-halle.de.
authors.forename: C Douglas
authors.initials: CD
authors.lastname: Grubb
authors.affiliation: Department of Molecular Signal Processing, Leibniz-Institute of Plant Biochemistry, Weinberg 3, D-06120 Halle, Germany. Electronic address: steffen.abel@ipb-halle.de.
authors.forename: Steffen
authors.initials: S
authors.lastname: Abel
mesh_terms: D000040:Abscisic Acid / Q000032:analysis; D000598:Amino Acids, Cyclic / Q000032:analysis*; D017360:Arabidopsis / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003517:Cyclopentanes / Q000032:analysis; D005449:Fluorenes / Q000737:chemistry*; D007210:Indoleacetic Acids / Q000032:analysis; D054883:Oxylipins / Q000032:analysis; D010937:Plant Growth Regulators / Q000032:analysis*; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014026:Nicotiana / Q000737:chemistry
chemical_list: D000598:Amino Acids, Cyclic; D003517:Cyclopentanes; D005449:Fluorenes; D007210:Indoleacetic Acids; D054883:Oxylipins; D010937:Plant Growth Regulators; C023863:1-aminocyclopropane-1-carboxylic acid; C011006:jasmonic acid; C030737:indoleacetic acid; D000040:Abscisic Acid; C054007:1-(9-fluorenyl)methyl chloroformate
keywords: 1-Aminocyclopropan-1-carboxylic acid; ACC; LC–MS/MS; Plant hormone profiling