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Mass spectrometry is a sensitive and specific method of proteome analysis. Various strategies are applied to increase the ionization efficiency of peptides and determine or compare their concentrations in complex samples, involving modifications introducing fixed charge for signal intensification and isotopic labeling for quantitative analysis. Scientific concepts involved in design and optimization of peptide markers, including solid phase synthesis and isotopic exchange are discussed. From the challenges related to the analysis of post-translational modifications emerge the perspectives for the development and validation of efficient derivatization reagents.

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