RESUMEN
A method for the chiral separation of dansyl-DL-amino acid mixtures using a dextran (M(r) 2,000,000) polymer network containing beta-cyclodextrin (beta-CD) in HPCE was developed. Mixtures of up to seven amino acids could be baseline resolved by this method under neutral pH conditions. Resolution was dependent on the dextran concentration in the polymer network. Temperature effects on the chiral separation were studied. Optimal efficiency and resolution of DL-enantiomeric pairs of amino acid samples were obtained at 25 degrees C. The resolution of different amino acids used in this work decreased with an increase in temperature.
Asunto(s)
Aminoácidos/aislamiento & purificación , Ciclodextrinas/química , Compuestos de Dansilo/aislamiento & purificación , Dextranos/química , Electroforesis/métodos , Polímeros , beta-Ciclodextrinas , Estereoisomerismo , TemperaturaRESUMEN
Chromatographic retention parameters of a series of 7 beta-adrenolytics and of 12 antihistamine drugs were determined employing an alpha 1-acid glycoprotein (AGP) high-performance liquid chromatographic (HPLC) column. For the group of antihistamines capillary electrophoretic (CE) retention was additionally measured in the presence of either AGP or human serum albumin (HSA). Two series of solutes hydrophobicity parameters were obtained by reversed-phase HPLC on an immobilized artificial membrane (IAM) column. The solutes studied were subjected to molecular modelling and the structural descriptors obtained were applied in studies of quantitative structure-retention (protein binding) relationships (QSRR). It was found that retention on AGP correlates well with the literature on physiological protein binding data. This retention was demonstrated to depend on hydrophobicity: to a lesser extent in the case of beta-adrenolytics and strongly in the case of antihistamines. Hydrophobicity, along with molecular width and electron excess charge on aliphatic nitrogen was demonstrated to describe retention of antihistamines on AGP. The AGP column is recommended as a convenient reactor for studies of drug-protein interactions. Preliminary CE data do not correlate with the HPLC data.
Asunto(s)
Antagonistas Adrenérgicos beta/química , Cromatografía Líquida de Alta Presión , Antagonistas de los Receptores Histamínicos H1/química , Orosomucoide/metabolismo , Antagonistas Adrenérgicos beta/metabolismo , Sitios de Unión , Electroforesis , Antagonistas de los Receptores Histamínicos H1/metabolismo , Metoprolol , Pindolol , Análisis de Regresión , Relación Estructura-ActividadRESUMEN
A method for the determination of the (6R)- and (6S)-stereoisomers of leucovorin using electrokinetic chromatography (EKC) in the affinity mode has been developed. Bovine serum albumin (BSA) is used as a run buffer additive to incorporate enantiomeric selectivity into the system. Protein-wall interactions are minimized by using a poly(ethylene glycol) (PEG) coated capillary. Chiral resolution is obtained in 12.5 min with efficiencies greater than 200,000 theoretical plates using BSA as an additive, while no resolution is obtained in the absence of BSA. A general equation is derived to calculate the free energy of interaction between the leucovorin isomers and the BSA molecule. This method represents a new means of obtaining thermodynamic data for substrate binding interactions and for the general study of drug cross-reactions and interactions of drugs with serum and other proteins.