Measurement of cyclosporin A and of four metabolites in whole blood by high-performance liquid chromatography.

A high-performance liquid chromatographic procedure using cyclosporin D as internal standard for the routine measurement of cyclosporin A and four of its metabolites is described. Whole-blood samples were purified on refillable solid-phase glass extraction columns. The chromatographic method includes a gradient elution using acetonitrile and water (pH 3.0) as eluents and an RP-8 analytical column. More than 1000 samples have already been analysed without any loss. The inter-assay variation was 6.3% and the intra-assay variation 4.9%. A linear correlation was found over a range of 0-3000 ng cyclosporin A per ml whole blood. The detection limit was 20 ng and the recovery was found to be 80-90%. Metabolites 1, 17, 18 and 21 could be characterized.