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1P-09 PDF
質量分析計を用いるペプチド分析に適したHPLC条件の検討
HPLC is widely used for analysis of peptides and the solutions containing trifluoroacetic acid (TFA) is generally used because of excellent peak shape and recovery rate. But, especially in the case of electrospray ionization, TFA reduces the sensitivity of analytes by signal suppression effect.
In this study, various mobile phase conditions were investigated to establish analytical method using mass spectrometry of peptides with high sensitivity, good peak shape and recovery rate. 8 kind of organic acids, which were C1-C4 and perfluorinated or partially fluorinated organic acids, were examined as additives in mobile phase. The separation of the peptides was performed on various analytical columns. The obtained chromatogram were evaluated on the basis of the retention factor, symmetry factor and recovery rate.
Signal intensity was reduced by using solution containing perfluorinated organic acid. The organic acid with high incompletion of fluorination showed a smaller signal suppression effect. It was confirmed that signal suppression effect was influenced by the state of the droplet due to the surface tension depends on acidity more than the length of alkyl chain. As a result of examining the optimum conditions, it was suggested that triflouropropionic acid (TFPA) was suitable as additive.