title: Stereoselective separations of chiral phosphinic acid pseudodipeptides by CEC using silica monoliths modified with an anion-exchange-type chiral selector.
abstract: A nonaqueous CEC method for the simultaneous separation of the four stereoisomers of the N-benzyloxycarbonyl phosphinic pseudodipeptide methyl ester benzyloxycarbonyl-homophenylalanine Z-hPhepsi(PO2HCH2)Phe-OCH3 as well as of the corresponding N-2,4-dinitrophenyl (DNP)-derivative with free C-terminal carboxylic group DNP-hPhepsi(PO2HCH2)Phe-OH was developed. For this purpose, a monolithic silica capillary column modified with a cinchona alkaloid-derived anion-exchange-type chiral selector, namely O-9-(tert-butylcarbamoyl)quinidine (tBuCQD) was prepared. The mobile phase composition (ACN/methanol ratio, counterion type) was thoroughly optimized to end up with baseline resolution of all four stereoisomers with critical resolution of as high as about 2. The CEC method proved to be superior over the corresponding HPLC separations primarily due to significantly enhanced plate numbers (between 200,000 and 600,000 m(-1) in CEC). Diastereoselectivity contributions arising from electrophoretic mobility differences of the diastereomers facilitated the separation of the later eluted diastereomeric peak pair (peaks III and IV), but had a negative influence on the selectivity of the earlier eluted diastereomeric peak pair (peaks I and II). The stereoselective CEC assay allowed the assessment of the stereoisomeric purity of the individual isomers which were obtained by preparative HPLC on a CHIRALPAK QD-AX column that is based on the same tBuCQD selector. The present study demonstrates that there exist problems which are hard to solve by HPLC, yet can be conveniently solved by CEC. Moreover, it was intended to prove by this practical application that CEC with monolithic columns is robust enough to be used for solving real-life problems.
authors.affiliation: Christian Doppler Laboratory for Molecular Recognition Materials, Department of Analytical & Food Chemistry, University of Vienna, Vienna, Austria.
authors.forename: Beatrix
authors.initials: B
authors.lastname: Preinerstorfer
authors.forename: Dieter
authors.initials: D
authors.lastname: Lubda
authors.forename: Artur
authors.initials: A
authors.lastname: Mucha
authors.forename: Paweł
authors.initials: P
authors.lastname: Kafarski
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Lindner
authors.forename: Michael
authors.initials: M
authors.lastname: Lämmerhofer
mesh_terms: D000837:Anion Exchange Resins; D053801:Capillary Electrochromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D004151:Dipeptides / Q000737:chemistry / Q000302:isolation & purification*; D010721:Phosphinic Acids / Q000737:chemistry* / Q000302:isolation & purification; D012822:Silicon Dioxide / Q000737:chemistry; D013237:Stereoisomerism
chemical_list: C516548:2,4-dinitrophenyl-homophenylalanyl-psi(PO2HCH2)-phenylalanine; D000837:Anion Exchange Resins; D004151:Dipeptides; D010721:Phosphinic Acids; C516547:benzyloxycarbonyl-homophenylalanyl-psi(PO2HCH2)-phenylalanine methyl ester; D012822:Silicon Dioxide