High-performance liquid chromatographic determination of the novel antitumour drug topotecan and topotecan as the total of the lactone plus carboxylate forms, in human plasma

摘要: A sensitive high-performance liquid chromatographic (HPLC) assay has been developed and validated for the quantitation of novel anticancer agent topotecan as total its lactone carboxylate forms in human plasma. Linear response analyte standard peak area were observed over concentration range 0.05–10 ng/ml using 100-μl plasma samples. The instability drug biological matrix necessitated that fraction was obtained within 5 min after blood sampling by centrifugation, immediately followed protein precipitation with cold methanol (−30°C). Stability studies have indicated is stable these methanolic extracts at least 4.5 months −30°C 2 −70°C. For determination plus ring-opened topotecan, samples deproteinated and, subsequently, acidified 7% (v/v) perchloric acid. Plasma measurement levels when stored −30°C. After supernatants analysed HPLC a Zorbax SB-C18 Stable Bond column methanol-0.1 M hexane-1-sulfonic acid methanol-0.01 N,N,N′,N′-tetramethylethylenediamine (TEMED) distilled water pH 6.0 (25:10:65, v/v) mobile phase. Detection performed fluorimetrically. Within-run between-run precision always less than 12.1% interest (0.05–10.0 ng/ml). limit 0.05 ng/ml. Accuracy measurements ranged between 87.6 113.5%.