RESUMO
The need for increased throughput in the quantitation of target compounds in biological fluids by high performance liquid chromatography/mass spectrometry continues to drive research in this area. This report describes the application of a prototype dual sprayer electrospray source for the quantitative analysis of biological samples. Quantitative performance for 180 compounds in a microsomal stability assay was found to be adequate when compared with a conventional single sprayer measurement. Issues with use of dual sprayers in a routine production environment are discussed.
Assuntos
Espectrometria de Massas por Ionização por Electrospray/instrumentação , Espectrometria de Massas por Ionização por Electrospray/métodos , Animais , Fatores Biológicos/análise , Fatores Biológicos/química , Fatores Biológicos/metabolismo , Cromatografia Líquida de Alta Pressão/instrumentação , Cromatografia Líquida de Alta Pressão/métodos , Meia-Vida , Microssomos Hepáticos/química , Microssomos Hepáticos/metabolismoRESUMO
Over the course of the last three or four years, the need for increased throughput analytical methods in support of drug discovery operations has dramatically increased. Once a nicety, methods which can provide reliable quantitation of target analytes in biological fluids are now necessary to keep pace with biological screening. To date, the only successful approaches to direct high-speed quantitation of large numbers of diverse compounds are based on high-performance liquid chromatography/mass spectrometric (HPLC/MS/MS) techniques. Key challenges to this task include sample preparation, the automated development of methods and the rapid analysis of many samples. Accomplishing these tasks requires the use of generic methods, which can be applied across many structurally diverse compounds. In this review, the state-of-the-art in high-speed HPLC/MS/MS is reviewed with the aim of describing the most practical and effective approaches currently in use.
RESUMO
An improved HPLC method for the analysis of 7-ethoxycoumarin and its metabolites in liver-slice incubation media is presented. The assay is based on short-column gradient elution which permits baseline resolution of the parent drug and three metabolites with a fourfold reduction in analysis time relative to existing HPLC methods. Total assay run time is 10 min. Data establishing method performance are presented which show intra- and interassay precision and accuracy to be within 13 and 7%, respectively, for separately prepared quality control samples.
