Expanding the range of sub/supercritical fluid chromatography: Advantageous use of methanesulfonic acid in water-rich modifiers for peptide analysis

The aim of this work was to expand the applicability range UHPSFC series synthetic and commercialized peptides. Initially, a screening different column chemistries available for analysis performed, in combination with additives either basic or acidic nature. an additive (13 mM TFA) stationary phase (Torus DEA 2-PIC) found be best six peptides possessing acidic, neutral isoelectric points. Secondly, methanesulfonic acid (MSA) evaluated as potential replacement TFA. Due its stronger acidity, MSA gave better performance than TFA at same concentration level. Furthermore, use reduced percentages MSA, such 8 mM, yielded similar results those observed 15 MSA. optimized method was, then, used compare against RP-UHPLC pI increasing peptide chain length. give slightly separation according their values, few cases orthogonal that UHPLC. On other hand, produced much increased amino compared Subsequently, UHPSFC-MS systematically UHPLC-MS using set linear cyclic commercially available. able generate least similar, some even UHPLC advantage providing complementary information given by analysis. Finally, analytical transferred semipreparative scale proprietary peptide, demonstrating excellent purity high yield less min.