Comparison of different electrokinetic separation modes applicable to a model peptide mixture (collagen type I and III CNBr fragments)

A number of electromigration separation modes were applied to the separation of CNBr-released peptides from rat tail tendon collagen (microemulsion electrokinetic chromatography, methanol- or ethanol-modified background electrolytes and the separation in the presence of molecular sieving effect exerting polymer, both in the presence and absence of SDS). Electrodriven separations with a Hypersil C8 packed capillary were investigated as well. The best separations were obtained with either the molecular sieving effect exerting polymer (polyethylene oxide) in the background electrolyte (whether SDS was present or absent) or with the electrodriven chromatography using the C8 reversed-phase packed capillary. In the latter separation system, it was possible to separate 25–27 peaks of the theoretically expected 24 peptides in the analyzed mixture of which 17 were at least tentatively identified. The additional peaks apparently stem from the incomplete cleavage of the parent collagen α chains. Successful separations can be done either with predominating molecular sieving or hydrophobic partitioning mechanism.