RESUMO
The relatively new field of combinatorial chemistry has enabled researchers to create large mixtures of compounds that can be screened for leads in developing potential drug candidates. The new synthetic method has also created a need for better procedures to analyze the complex mixtures that are generated. The immediate goal in most cases is to verify the synthetic procedure and to determine the purity and completeness of the library sample before binding studies are initiated. We report here a method to rapidly characterize small-molecule combinatorial libraries in solution. All combinatorial library samples were synthesized by combining a core molecule bearing two acid chloride functionalities with various amino acids to generate libraries of 36, 78 and 120 components. Using electrospray ionization fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR-MS) we were able to identify 70-80% of the library components. All samples were analyzed as mixtures by direct infusion without chromatographic separation. Furthermore, nominally isobaric components could be resolved and identified through exact mass assignments without tandem mass spectrometery. ESI-FTICR-MS is a rapid and convenient tool for the characterization of small-molecule libraries. The method is especially useful for the analysis of larger libraries that contain many nominally isobaric components and impurities.
Assuntos
Desenho de Fármacos , Biblioteca de Peptídeos , Química Farmacêutica , Ciclotrons , Espectrometria de Massas , Modelos Químicos , Espectroscopia de Infravermelho com Transformada de FourierAssuntos
Espectrometria de Massas/métodos , Sequência de Aminoácidos , Animais , Bovinos , Análise de Fourier , Cavalos , Insulina/química , Espectrometria de Massas/instrumentação , Espectrometria de Massas/estatística & dados numéricos , Dados de Sequência Molecular , Estrutura Molecular , Mioglobina/química , Fragmentos de Peptídeos/química , Sensibilidade e Especificidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodosRESUMO
A method has been demonstrated for producing accurate, reproducible gradients at low microliter per minute flow-rates suitable for liquid chromatographic-mass spectrometric (LC-MS) applications using capillary chromatography columns. The technique employs a low-cost, well characterized balance-column flow-splitter which can be simply added to a high-performance LC-MS solvent delivery system. Performance of the pump for LC-MS techniques at higher flow-rates is preserved.
