Biologically active peptides harbor an important role in human health. They are associated with the development of new therapeutic health care products and as potential biologic pathway markers and metabolites in the detection of disease. As such, we have chosen peptides that increase in amino acid length to highlight how the PRP-3 (300 Å) HPLC stationary phase is a useful tool for the field of peptides. Analysis of the stationary phase indicates good separation between Angiotensin I and II biomarkers, while further providing baseline resolution between the remaining analytes. The chromatogram also presents the peak shape and resolution an analyst can expect when utilizing the PRP-3 stationary phase. We have chosen in this example the 7 µm particle for its minimal system back pressure (550 psi) with the resolution needed for isolation in both an analytical analysis and a scale up processes.

The PRP-3 is the perfect complement to any peptide isolation. The 300 Å particle pore size helps optimize the interaction of larger molecular weight peptides with the stationary phase while minimizing clogged pores. After analytical isolation is achieved, scale up is where the PRP-3 excels due to its ability to accommodate ~20% more sample loading before observing column overload compared to non- and superficially-porous stationary phases. Loading capacity is further optimized due to the absence of analyte absorption sites commonly found in silica ODS stationary phases like, acidic silanol groups. Despite lacking a silica core, the PRP-3 PSDVB particle provides the consistency and excellence that customers have come to expect from all Hamilton products.

Author: Adam L. Moore, PhD, Hamilton Company

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Separation of Biologically Active Increasing Length Peptides (PRP-3)