Quantification of Phytochelatins in Chlamydomonas reinhardtii using ferrocene-based derivatization

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.