Peptide separations and dissociation constants in nonaqueous capillary electrophoresis: comparison of methanol and aqueous buffers.
Nonaqueous capillary electrophoresis was evaluated for its potential to separate peptides in methanolic background electrolytes in comparison to aqueous-methanol (50% v/v) and water. Isomeric aspartyl dipeptides and Leu- and Met-enkephalin served as model compounds. pK(a) values were determined in the three solvent systems based on the apparent pH scale and in the case of methanol additionally based on the conventional pH scale. Changing from water to methanol led to an increase of the ionization constants describing the dissociation equilibria of the carboxyl group and the amino group, respectively. The pK(a) shift was more pronounced for the carboxylic acid function leading to a compression of the mobility-pH curve. As reported for aqueous buffers, efficient separations of the peptides were achieved in methanolic background electrolytes including the resolution of the diastereomers of the isomeric alpha- and beta-aspartyl dipeptides. In contrast to aqueous buffers, the separation of Leu- and Met-enkephalin could also be obtained in buffers in methanol at high pH.
Psurek
Arndt
A
University of Jena, School of Pharmacy, Department of Pharmaceutical Chemistry, Jena, Germany.
Scriba
Gerhard K E
GK
D002021:Buffers; D004573:Electrolytes; D019075:Electrophoresis, Capillary / Q000379:methods*; D004743:Enkephalin, Leucine / Q000032:analysis / Q000737:chemistry; D004744:Enkephalin, Methionine / Q000032:analysis / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008433:Mathematics; D000432:Methanol / Q000737:chemistry; D008956:Models, Chemical; D009479:Neuropeptides / Q000032:analysis* / Q000737:chemistry; D010455:Peptides / Q000032:analysis* / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D013237:Stereoisomerism
D002021:Buffers; D004573:Electrolytes; D009479:Neuropeptides; D010455:Peptides; D012997:Solvents; D004744:Enkephalin, Methionine; D004743:Enkephalin, Leucine; D000432:Methanol