Final answer:
Peptides can be separated using an ion-exchange column that separates them based on their net charge, with the ability to adjust pH and ionic strength to fine-tune separation. Other HPLC techniques, such as reverse-phase HPLC, leverage hydrophobic interactions for peptide separation before mass spectrometry analysis.
Step-by-step explanation:
Yes, peptides can be separated using an ion-exchange column which is a form of chromatography that uses a column with a stationary phase that will bind to charged peptide species. In ion-exchange chromatography, peptides are separated based on their net charge at the pH of the mobile phase. By adjusting the pH and ionic strength of the buffers used to elute peptides from the column, it is possible to influence the interaction between the peptides and the stationary phase, thereby separating the peptides based on their charge properties. Additionally, high-performance liquid chromatography (HPLC) techniques, such as reverse-phase HPLC, can be employed for the separation of peptides after digestion and before MS analysis. In reverse-phase HPLC, peptides are typically separated based on hydrophobic interactions between the peptides and the nonpolar stationary phase of the column.
Another method detailed in the provided text includes the use of Ultra Performance Liquid Chromatography (UPLC) followed by a low pH reverse-phase chromatography for the separation of tryptic peptides, which then are analyzed by mass spectrometry. Reverse-phase HPLC and UPLC techniques utilize columns packed with silica particles modified with alkane chains which interact with the hydrophobic regions of peptides. The degree of hydrophobicity affects the retention time of each peptide, allowing for their separation based on the differences in hydrophobic interactions