Improved and simplified liquid chromatographic assay for adefovir, a novel antiviral drug, in human plasma using derivatization with chloroacetaldehyde

摘要

A rapid and simplified chromatographic assay is reported for the quantification of adefovir (PMEA) utilizing derivatization with chloroacetaldehyde. Adefovir is isolated from plasma using protein preipitation with trichloroacetic acid: next, the fluorescent 1.N~6 -etheno derivative is directly formed at 98degC in the buffered extract with chloroacetaldehyde. This derivative is analyzed using isocratic ion-pair liquid chromatography and fluorescence detection at 254 nm for excitation and 425 nm for emission. In the evaluated concentration range (10-100 ng/ml) precisions <=5% and accuracies between 95 and 117% were found, using a 0.2 -ml volume of plasma. The lower limit of quantification is 10 ng/ml with a intra-assay precision of 16%.The currently reported bioanalytical method is 20-25-fold sensitive than previously published assays. 1999 Elsevier Science B.V. All rights reserved.