Enantiomer resolution in immobilized pH gradient gels via inclusion of a chiral discriminator.
The separation of enantiomeric forms of dansylated amino acids by isoelectric focusing in immobilized pH gradients (IPG) is demonstrated for the first time. Separations occur in a pH 3.0-4.0 IPG interval, in presence of 7Murea, 10% methanol and 60 mM beta-cyclodextrin (CD) as chiral discriminator. It is found that the inclusion complex formed between the D-form and CD has a lower pI than the uncomplexed form (delta pI = 0.05 for DL-Phe and delta pI = 0.025 for DL-Trp); from this, it is calculated that the pK of the tertiary amino group in the dansyl moiety is lowered by 0.1 pH unit in the former case (D-Phe) and by 0.05 in the case of D-Trp (both values referring to 60 mM CD gels). For some racemates (e.g., DL-Phe) the separation mechanism is still operative with CD concentrations as low as 20 mM. In our system 60 mM CD appears to be the solubility limit of CD. As the complex is stable in the electric field for at least 15 h, this separation mechanism could be exploited for purifying large quantities of pure D and L forms from racemates in multicompartment electrolyzers with isoelectric Immobiline membranes.
Righetti
P G
PG
Department of Biomedical Sciences and Technologies, University of Milano, Italy.
Ettori
C
C
Chafey
P
P
Wahrmann
J P
JP
D000596:Amino Acids / Q000302:isolation & purification*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003505:Cyclodextrins; D003619:Dansyl Compounds / Q000302:isolation & purification*; D003912:Dextrins; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D010649:Phenylalanine / Q000302:isolation & purification; D013213:Starch; D013237:Stereoisomerism; D014364:Tryptophan / Q000302:isolation & purification; D047392:beta-Cyclodextrins
D000596:Amino Acids; D003505:Cyclodextrins; D003619:Dansyl Compounds; D003912:Dextrins; D047392:beta-Cyclodextrins; D010649:Phenylalanine; D014364:Tryptophan; D013213:Starch; C031215:betadex