Introduction to Peptide Analysis by HPLC

The high resolving power of reversed phase HPLC make it a powerful tool when analyzing peptides digested from larger proteins. This webcast will introduce the process by which the parameters of reversed phase HPLC can be optimized to analyze peptides; we will consider the stationary phase chemistry which impacts selectivity, as well as the impact the physical attributes of the column have on peptide separations. Furthermore, modification of the mobile phase using TFA, formic acid, and pH will be discussed in relation to chromatographic results and MS compatibility.

Topics include:

• Introduction to peptide synthesis
• Outline of regulatory guidelines
• Amino acid, peptide, and protein chemistry
• Reversed phase retention mechanism for biomolecules
• Phase selectivity
• Pore size
• Particle type
• Mobile phase conditions
• Compatibility with MS