Analytical and Bioanalytical Chemistry

, Volume 398, Issue 2, pp 877–883

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

  • Anja Bräutigam
  • Dirk Wesenberg
  • Hugues Preud’homme
  • Dirk Schaumlöffel
Original Paper

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, PC2, PC3, CysPC2, and CysPC3, were quantified with high reproducibility at concentrations between 15 and 198 nmol g−1 fresh weight. The biological variation of PC synthesis of nine independently grown alga cultures was determined to be on average 13.7%.

Figure

A rapid UPLC-MS/MS method was developed for thiol peptide quantification in micro litre sample volumes at the nanomol level. For the first time reproducible quantification of six thiol peptides (GSH, CysGSH, PC2, PC3, CysPC2 and CysPC3) in crude extracts of Clamydomonas reinhardtii was possible

Keywords

Phytochelatin quantification Chlamydomonas UPLC ESI-MS/MS 

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Anja Bräutigam
    • 1
    • 3
  • Dirk Wesenberg
    • 1
  • Hugues Preud’homme
    • 2
  • Dirk Schaumlöffel
    • 2
  1. 1.Martin-Luther-Universität Halle-Wittenberg, Institut für Biochemie und BiotechnologieÖkologische und Pflanzen-BiochemieHalle (Saale)Germany
  2. 2.Université de Pau et des Pays de l’Adour/CNRS UMR 5254Laboratoire de Chimie Analytique Bio-Inorganique et Environnement/IPREMPauFrance
  3. 3.Landeskriminalamt Sachsen-AnhaltMagdeburgGermany

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