Quantification of Phytochelatins in Chlamydomonas reinhardtii using ferrocene-based derivatization

Anja Bräutigam; Susanne Bomke; Thorben Pfeifer; Uwe Karst; Gerd-Joachim Krauss; Dirk Wesenberg

doi:10.1039/C005014H

Quantification of Phytochelatins in Chlamydomonas reinhardtii using ferrocene-based derivatization†

Anja Bräutigam,^a Susanne Bomke,^b Thorben Pfeifer,^b Uwe Karst,^b Gerd-Joachim Krauss^a and Dirk Wesenberg*^a

* Corresponding authors

^a Martin-Luther-Universität Halle-Wittenberg, Institut für Biochemie und Biotechnologie, Ökologische und Pflanzen-Biochemie, Kurt-Mothes-Straße 3, 06120 Halle (Saale), Germany
E-mail: dirk.wesenberg@biochemtech.uni-halle.de
Fax: +49 3455527012

^b Westfälische Wilhelms-Universität Münster, Institut für Anorganische und Analytische Chemie, Corrensstrasse 30, Münster, Germany

Abstract

A method for the identification and quantification of canonic and isoforms of phytochelatins (PCs) from Chlamydomonas reinhardtii was developed. After disulfide reduction with tris(2-carboxyethyl)phosphine (TCEP) PCs were derivatized with ferrocenecarboxylic acid (2-maleimidoyl)ethylamide (FMEA) in order to avoid oxidation of the free thiol functions during analysis. Liquid chromatography (LC) coupled to electrospray mass spectrometry (ESI-MS) and inductively coupled plasma-mass spectrometry (ICP-MS) was used for rapid and quantitative analysis of the precolumn derivatized PCs. PC₂₋₄, CysGSH, CysPC₂₋₄, CysPC₂desGly, CysPC₂Glu and CysPC₂Ala were determined in the algal samples depending on the exposure of the cells to cadmium ions.

Metallomics

Quantification of Phytochelatins in Chlamydomonas reinhardtii using ferrocene-based derivatization†

Abstract

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