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Schmidt, Holger ; Böttcher, Christoph ; Trampczynska, Aleksandra ; Clemens, Stephan:
Use of recombinantly produced 15N3-labelled nicotianamine for fast and sensitive stable isotope dilution ultra-performance liquid chromatography/electrospray ionization time-of-flight mass spectrometry.
In: Analytical and Bioanalytical Chemistry.
Bd. 399
(2011)
Heft 3
.
- S. 1355-1361.
ISSN 1618-2650
DOI: https://doi.org/10.1007/s00216-010-4436-7
Abstract
Nicotianamine (NA) is an important metal chelator, implicated in the intra- and intercellular trafficking of several transition metal ions in plants. To decipher its roles in physiological processes such as micronutrient acquisition, distribution or storage, fast and sensitive analytical techniques for quantification of this non-proteinogenic amino acid will be required. The use of a recombinant Schizosaccharomyces pombe strain expressing a nicotianamine synthase (NAS) gene allowed for the production of [15N3]-NA, which was enriched from cell extracts through cation exchange and used for stable isotope dilution analysis of NA. Such an approach should be widely applicable to important bioanalytes that are difficult to synthesize. The analytical procedure comprises mild aqueous extraction and rapid Fmoc derivatization, followed by fast separation using ultra-performance liquid chromatography (UPLC) and sensitive detection by positive ion electrospray ionization time-of-flight mass spectrometry (ESI-TOF-MS) with a chromatographic cycle time of only 8 min. Derivatization was optimized with respect to incubation time and species suitable for quantification. Limit of detection was 0.14 to 0.23 pmol in biological matrices with the response being linear up to 42 pmol. Recovery rates were between 83 and 104 % in various biological matrices including fission yeast cells, fungal mycelium, plant leaves and roots.
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| Publikationsform: | Artikel in einer Zeitschrift |
|---|---|
| Begutachteter Beitrag: | Nein |
| Zusätzliche Informationen: | BAYCEER90325 |
| Institutionen der Universität: | Fakultäten > Fakultät für Biologie, Chemie und Geowissenschaften > Fachgruppe Biologie > Lehrstuhl Pflanzenphysiologie Fakultäten > Fakultät für Biologie, Chemie und Geowissenschaften > Fachgruppe Biologie > Lehrstuhl Pflanzenphysiologie > Lehrstuhl Pflanzenphysiologie - Univ.-Prof. Dr. Stephan Clemens Forschungseinrichtungen > Forschungszentren > Bayreuther Zentrum für Ökologie und Umweltforschung - BayCEER Fakultäten Fakultäten > Fakultät für Biologie, Chemie und Geowissenschaften Fakultäten > Fakultät für Biologie, Chemie und Geowissenschaften > Fachgruppe Biologie Forschungseinrichtungen Forschungseinrichtungen > Forschungszentren |
| Titel an der UBT entstanden: | Ja |
| Themengebiete aus DDC: | 500 Naturwissenschaften und Mathematik |
| Eingestellt am: | 29 Apr 2015 15:42 |
| Letzte Änderung: | 29 Apr 2015 15:42 |
| URI: | https://eref.uni-bayreuth.de/id/eprint/11705 |