RESUMO
A sensitive method for GC-ECD simultaneous determination of nitrendipine and its pyridine metabolite M1 in human plasma is described. Felodipine was used as the internal standard. The plasma samples were extracted with toluene. One microlitre of the extract was injected onto the capillary column (polymethylsiloxane) and measured with electron-capture detector. The developed method showed to be linear over the range 0.25-70 for nitrendipine and 0.3-61 ng/ml for its metabolite M1 with an inter-day and intra-day precision in terms of R.S.D. lower than 8% except the concentrations near lowest limit of quantification (LLOQ) (<11% R.S.D.). The LLOQ for nitrendipine was 0.25 and 0.3 ng/ml for its metabolite, respectively. The analytical recovery was 94% for nitrendipine and 89% for its pyridine metabolite M1. This GC-ECD method was developed for being used in clinical pharmacokinetic studies.
Assuntos
Cromatografia Gasosa/métodos , Nifedipino/sangue , Nifedipino/metabolismo , Calibragem , Estabilidade de Medicamentos , Felodipino/sangue , Felodipino/química , Congelamento , Humanos , Estrutura Molecular , Nifedipino/química , Piridinas/sangue , Piridinas/metabolismo , Padrões de Referência , Sensibilidade e Especificidade , Fatores de TempoRESUMO
A new bioanalytical high-performance liquid chromatographic (HPLC) method for the determination of ciprofloxacin with norfloxacin as an internal standard was developed and validated for plasma samples. Norfloxacin is structural homologue of ciprofloxacin and exhibits similar retention properties. The quality of respective peak separation is strongly influenced by amphoteric character of ciprofloxacin and norfloxacin as well. In previously published HPLC methods on conventional C18 reversed-phase [F. Belal, A.A. Al-Majed, A.M. Al-Obaid, Talanta 50 (1999) 765-786; G. Carlucci, J. Chromatogr. A 812 (1998) 343-367], ion pair reagents were added into the mobile phase to suppress peak tailing. In comparison with endcapped and high purity silica reversed-phase sorbent (Purospher RP-18e, Merck), which yielded symmetrical peaks, separation efficiency was further enhanced in our method. Gradient elution mode using acetonitrile and phosphate buffer pH 3 on the pentafluorophenylpropyl stationary phase (250-4.6 mm Discovery HS F5, 5 microm, Supelco) was carried out. The resolution of 4.1 for ciprofloxacin-norfloxacin peaks was achieved. Sample preparation by SPE C18 (Supelclean) with recovery 72% was performed. Fluorescence detection with lambda(excit)=280 nm, lambda(emis)=446 nm was used. After the validation, the bioanalytical HPLC method was applied to pharmacokinetic studies.
