EVALUATION OF SOLID-PHASE EXTRACTION PROCEDURES IN PEPTIDE ANALYSIS

Solid-phase extraction (SPE) procedures for peptide isolation and fractionation, based on non-polar and ionic interactions, were evaluated using small synthetic peptides and casein enzymatic hydrolysates. SPE based on hydrophobic phases is a useful, efficient and rapid procedure for peptide extraction and concentration. It allows a successful peptide fractionation using eluents that contain an increasing content of acetonitrile in the presence of trifluoroacetic acid. Differences regarding selectivity are observed between sorbents. Non-polar interaction with C-18 sorbents is adequate for the isolation of very polar and hydrophobic peptides. CN sorbents are only adequate for very hydrophobic peptides, PH, CH, C-8 and C-2 sorbents are useful for isolating and fractionating hydrophobic and very non-polar peptides, but generally not for very polar peptides. Ionic solid-phase extraction using Accell Plus cartridges of QMA (quaternary methylammonium) and CM (carboxymethyl) are very useful for the fractionation of peptide mixtures into basic, acidic and neutral pools of peptides. It can be concluded that SPE using these procedures is a useful tool for the isolation and fractionation of peptides from biological and food samples.