TITLE

Rapid and simple UPLC-MS/MS method for precise phytochelatin quantification in alga extracts

AUTHOR(S)
Bräutigam, Anja; Wesenberg, Dirk; Preud'homme, Hugues; Schaumlöffel, Dirk
PUB. DATE
September 2010
SOURCE
Analytical & Bioanalytical Chemistry;Sep2010, Vol. 398 Issue 2, p877
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Quantitative phytochelatin (PC) analysis is, due to oxidation sensitivity of the PCs, matrix effects, and time consuming sample preparation, still a challenging analytical task. In this study, a rapid, simple, and sensitive method for precise determination of native PCs in crude extracts of the green alga Chlamydomonas reinhardtii was developed. Algae were exposed 48 h to 70 μM Cd. Coupling of ultra performance liquid chromatography and electrospray ionization tandem mass spectrometry with multi-reaction mode transitions for detection permitted the required short-time, high-resolution separation and detection specificity. Thus, under optimized chromatographic conditions, 10 thiol peptides were baseline-separated within 7 min. Relative detection limits in the nanomolar range in microliter sample volumes were achieved (corresponding to absolute detection limits at femtomol level). Next to glutathione (GSH), the most abundant cadmium-induced PCs in C. reinhardtii, namely CysGSH, PC, PC, CysPC, and CysPC, were quantified with high reproducibility at concentrations between 15 and 198 nmol g fresh weight. The biological variation of PC synthesis of nine independently grown alga cultures was determined to be on average 13.7%. [Figure not available: see fulltext.]
ACCESSION #
53556233

 

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