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

Anja Bräutigam, Dirk Wesenberg, Hugues Preud'Homme, Dirk Schaumlöffel

Research output: Contribution to journalArticle

28 Citations (Scopus)

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 2, 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 not available: see fulltext.]

Original languageEnglish
Pages (from-to)877-883
Number of pages7
JournalAnalytical and Bioanalytical Chemistry
Volume398
Issue number2
DOIs
Publication statusPublished - 1 Sep 2010
Externally publishedYes

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Phytochelatins
Algae
Chlamydomonas reinhardtii
Limit of Detection
Electrospray ionization
Chlorophyta
Electrospray Ionization Mass Spectrometry
Liquid chromatography
Tandem Mass Spectrometry
Complex Mixtures
Cadmium
Sulfhydryl Compounds
Liquid Chromatography
Mass spectrometry
Glutathione
Weights and Measures
Oxidation
Peptides
Chemical analysis

Keywords

  • Chlamydomonas
  • ESI-MS/MS
  • Phytochelatin quantification
  • UPLC

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

Cite this

Rapid and simple UPLC-MS/MS method for precise phytochelatin quantification in alga extracts. / Bräutigam, Anja; Wesenberg, Dirk; Preud'Homme, Hugues; Schaumlöffel, Dirk.

In: Analytical and Bioanalytical Chemistry, Vol. 398, No. 2, 01.09.2010, p. 877-883.

Research output: Contribution to journalArticle

Bräutigam, Anja ; Wesenberg, Dirk ; Preud'Homme, Hugues ; Schaumlöffel, Dirk. / Rapid and simple UPLC-MS/MS method for precise phytochelatin quantification in alga extracts. In: Analytical and Bioanalytical Chemistry. 2010 ; Vol. 398, No. 2. pp. 877-883.
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