RESUMO
This paper describes the enantiomeric resolution of a series of unsaturated N-methyloxycarbonyl-alpha-H-alpha-amino acids (N-MOC-alpha-amino acids) on macrocyclic glycopeptide stationary phases by means of high-performance liquid chromatography (HPLC). Three types of glycopeptide phases, i.e. Chirobiotic T, V and R, were evaluated in both reversed-phase (RP) and polar ionic mode (PIM). The best results in terms of enantioselectivity and resolution were obtained on Chirobiotic R phase, with the PIM mobile phase giving the highest resolution per min. Investigation of the pH of the reversed-phase mobile phase in the pH range 4.1-5.9 showed little effect on enantioselectivity. The method was applied for monitoring the conversion and product enantiomeric excess of an enzymatic hydrolysis reaction using N-MOC-alpha-H-alpha-amino acid esters as substrate.
Assuntos
Aminoácidos/isolamento & purificação , Cromatografia Líquida de Alta Pressão/métodos , Glicopeptídeos/química , EstereoisomerismoRESUMO
o-Phthalaldehyde in combination with chiral thiols is described as a chiral reagent for the resolution of the enantiomers of alpha-H-alpha-aminonitriles and alpha-alkyl-alpha-aminonitriles. Separation of the resulting diastereomers was carried out by reversed-phase chromatography and the derivatives were detected at UV 340 nm. The identity of the diastereomeric derivatives was confirmed by LC-MS. Among the chiral thiols tested, N-acetyl-D-penicillamine and beta-mercaptoisobutyric acid gave the best resolution for the alpha-aminonitriles studied. For quantitative enantiomeric excess determination, beta-mercaptoisobutyric acid was chosen as this thiol could be obtained in high enantiomeric purity from a commercial source. The rate of the reaction of various alpha-aminonitriles with o-phthalaldehyde and beta-mercaptoisobutyric acid was studied. The accuracy of the method was investigated by a comparison of theoretical and measured enantiomeric excess.
