RESUMO
A novel, simple and mass spectrometry (MS) compatible high-performance liquid chromatography (HPLC) method is reported for the simultaneous estimation of ilaprazole (ILA) and glimepiride (GLM) in rat plasma. The bio-analytical procedure involves extraction of ILA, GLM and internal standard (IS) from rat plasma with a solid-phase extraction (SPE) process. The chromatographic analysis was performed on Waters-600 system using an isocratic mobile phase comprising methanol:water (80:20 % v/v) with pH of water modified to three using formic acid at a flow rate of 1.0 mL/min and Kinetex C18 column maintained at 30 ± 1°C. The signals were monitored using a PDA detector set at 225 nm. IS, ILA and GLM eluted at 2.04, 4.7 and 7.4 min, respectively, and the total run time was 10 min. Method validation was performed as per US Food and Drug Administration guidelines and the results met the acceptance criteria. The calibration curve was linear over a concentration range of 10-600 ng/mL (r2 = 0.999). The intra- and inter-day precisions for ILA and GLM were (%RSD values) in the range of 1.52-9.74 and 1.52-11.76%, respectively, in rat plasma. The method was successfully applied in pharmacokinetic studies followed by oral administration of GLM and ILA in rats.
Assuntos
2-Piridinilmetilsulfinilbenzimidazóis/sangue , Cromatografia Líquida de Alta Pressão/métodos , Extração em Fase Sólida/métodos , Compostos de Sulfonilureia/sangue , 2-Piridinilmetilsulfinilbenzimidazóis/química , 2-Piridinilmetilsulfinilbenzimidazóis/farmacocinética , Animais , Estabilidade de Medicamentos , Modelos Lineares , Masculino , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Compostos de Sulfonilureia/química , Compostos de Sulfonilureia/farmacocinéticaRESUMO
Present work describes the development and validation of a simple and reliable high-performance liquid chromatography-diode array detection (HPLC-DAD) procedure for the analysis of phenylephrine hydrochloride (PHE), paracetamol (PAR) and cetirizine dihydrochloride (CET), in pharmaceutical mixture. The method was applied successfully on tablet dosage form. Effective chromatographic separation of PHE, PAR and CET was achieved using a Kinetex-C18 (4.6, 150 mm, 5 mm) column with gradient elution of the mobile phase composed of 10 mM phosphate buffer (pH 3.3) and acetonitrile. The elution was a 3 step gradient elution program step-1 started initially with 2% (by volume) acetonitrile and 98% phosphate buffer (pH 3.3) for first 2 min. In step-2 acetonitrile concentration changed linearly to 20% upto 12 min the analysis was concluded by step-3 changing acetonitrile to 2% upto 20 min. The reliability and analytical performance of the proposed HPLC procedure were statistically validated with respect to linearity, ranges, precision, accuracy, selectivity and robustness. Calibration curves were linear in the ranges 5-15, 250-750 and 2.5-7.5 µg/ml for PHE, PAR and CET, with correlation coefficients >0.9996. The validated HPLC method was applied to a pharmaceutical mixture of a marketed preparation tablet in which the analytes were successfully quantified with good recovery values with no interfering peaks from the excipents.
Assuntos
Acetaminofen/química , Cetirizina/química , Fenilefrina/química , Comprimidos/química , Calibragem , Química Farmacêutica/métodos , Cromatografia Líquida de Alta Pressão/métodos , Excipientes/química , Reprodutibilidade dos TestesRESUMO
Sildenafil citrate (SIL) is used in the treatment of erectile dysfunction and other chronic disorders. For the pharmacokinetic investigation of SIL we developed a simple and sensitive method for the estimation of SIL in rat plasma by reverse phase high-performance liquid chromatography (RP-HPLC). The drug samples were extracted by liquid-liquid extraction with 300 µl of acetonitrile and 5 ml of diethyl ether. Chromatographic separation was achieved on C18 column using methanol:water (85:15 v/v) as mobile phase at a flow rate of 1 ml/min and UV detection at 230 nm. The retention time of SIL was found to be 4.0 min having a separation time less than 5 min. The developed method was validated for accuracy, precision, linearity and recovery. Linearity studies were found to be acceptable over the range of 0.1-6 µg/ml. The method was successfully applied for the analysis of rat plasma sample for the application in pharmacokinetic study, drug interaction, bioavailability and bioequivalence.
RESUMO
A simple and sensitive method was developed for simultaneous estimation of Glimepiride (GLIM) and Sildenafil citrate (SIL) in rat Plasma by reverse phase high performance liquid chromatography (RP-HPLC). The drug samples were extracted by liquid-liquid extraction with 300 µl of acetonitrile and 5 ml of diethyl ether. Chromatographic separation was achieved on C18 column using methanol: water (85:15 v/v) as mobile phase at a flow rate of 1 ml/min and UV detection at 230 nm. The retention time of GLIM and SIL was found to be 2.5 and 4.0 min respectively with total run time of 7 min. The developed method was validated for accuracy, precision, linearity and recovery. The method was linear and found to be acceptable over the range of 100-12 000 ng/ml. The method was successfully applied for the analysis of rat plasma sample for application to pharmacokinetic.
