UltraPerformance Convergence Chromatography (UPC2) offers an orthogonal choice to reversed phase liquid chromatography (RPLC) in DMPK applications, particularly in quantitative assays for bioanalysis.
One of the advantages of using UPC2 for bioanalysis is its compatibility with solvents used for sample preparation. Whether selecting solid-phase extraction, liquid-liquid extraction, or protein precipitation, the final extract is often dissolved in an organic solution that may not be compatible with RPLC without dilution. UPC2, however, is compatible with many organic solvents, eliminating the need for evaportation and reconstitution typically needed for RPLC.
UPC2 for bioanalylsis:
Using selectivity to avoid matrix effects
One method to avoid potential phospholipid coelution and therefore potential matrix effects is to change the chromatographic selectivity of a separation. In RPLC, relatively hydrophobic phospholipids require high percentages of organic solvent to elute from the column. Thus, hydrophobic compounds of interest are prone to coelute with these lipid interferences and experience matrix effects. UPC2 utilizes supercritical carbon dioxide as the major mobile phase, resulting in vast selectivity differences when compared to standard RPLC in bioanalysis.