Quantification of phytochelatins in plants by reversed-phase HPLC-ESI-MS-MS.
An on-line HPLC-ESI-MS-MS method has been developed for determination of glutathione and phytochelatins (PC) in plant tissues. For sample pretreatment, dithiothreitol (DTT) must be added at the very beginning, as an anti-oxidant. Optimization of instrumental conditions i.e. composition of HPLC mobile phase, ionization efficiency of the electrospray interface, and MS-MS detection in the multiple ion-monitoring mode, are the central aspects of this work. A polystyrene-packed column was found to be superior to a standard silica-packed reversed-phase column. A concave quadratic gradient of ammonium formate buffer and acetonitrile was found to be optimum. The limits of quantitation were 0.2 micromol kg(-1) plant tissue for glutathione and PC. The method has been applied to analysis of tissue samples from Vicia faba grown in Cd-containing nutrient solutions.
Environmental Chemistry and Ecotoxicology, University of Bayreuth, 95440 Bayreuth, Germany.
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El-Zohri
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Cabala
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Frank
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005978:Glutathione; D013058:Mass Spectrometry / Q000379:methods*; D008667:Metalloproteins / Q000032:analysis*; D054811:Phytochelatins; D018515:Plant Leaves / Q000737:chemistry; D010940:Plant Proteins / Q000032:analysis*; D018517:Plant Roots / Q000737:chemistry; D018629:Spectrometry, Mass, Secondary Ion / Q000379:methods*; D031307:Vicia faba
D008667:Metalloproteins; D010940:Plant Proteins; D054811:Phytochelatins; D005978:Glutathione