Abstract
A method based on ICP collision-cell MS detection in capillary HPLC was developed to gain an insight into the purity and identity of selenium-containing proteins separated by 1-D and 2-D electrophoresis. The bands and spots obtained after the separation of water-soluble proteins in selenized yeast were digested with trypsin prior to chromatography. Selenium could be detected down to the subpicogram level. The method, assisted by information obtained by MALDI TOF MS on the 5000 Da cut-off fraction, permitted the purity of bands and spots to be estimated and the efficiency of tryptic digestion and the quantity of selenium present in individual peptides to be evaluated. Owing to the high sensitivity and the lack of matrix suppression effects, the method provided chromatograms with signal-to-noise ratios of 10–1000 in conditions where the common ES Q–TOF MS detection failed.
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Acknowledgements
The authors thank Dr. R. Grimaud for his help with GE measurements and Dr. H. Preud’homme and Dr. O. Palacios for their help on the MALDI TOF MS. The financial support of the Aquitaine Region is acknowledged.
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Tastet, L., Schaumlöffel, D., Bouyssiere, B. et al. Capillary HPLC–ICP MS mapping of selenocompounds in spots obtained from the 2-D gel electrophoresis of the water-soluble protein fraction of selenized yeast. Anal Bioanal Chem 385, 948–953 (2006). https://doi.org/10.1007/s00216-006-0482-6
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DOI: https://doi.org/10.1007/s00216-006-0482-6