RESUMEN
A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for simultaneous quantification of Tenofovir (TEN) and Emtricitabine (EMT) in human plasma using Chromolith Speed Rod RP18. The mass transition ion-pair has been followed as m/z 288.10-->176.10 for TEN, m/z 248.20-->130.20 for EMT and m/z 230.10-->112.10 for Lamivudine (LAM). The method involves solid phase extraction from plasma, simple isocratic chromatographic conditions and mass spectrometric detection using an API 5000 instrument that enables detection at nanogram levels. Lamivudine was used as the internal standard. The proposed method has been validated with a linear range of 10-600 ng/ml for TEN and 25-2,500 ng/ml for EMT. The intrarun and interrun precision values are within 12.0% for TEN and 15.6% for EMT at their respective LOQ levels. The overall recoveries for TEN and EMT were 84.3% and 68.5%, respectively. Total elution time was as low as 2 min.
Asunto(s)
Adenina/análogos & derivados , Cromatografía Liquida/métodos , Desoxicitidina/análogos & derivados , Organofosfonatos/sangre , Inhibidores de la Transcriptasa Inversa/sangre , Espectrometría de Masas en Tándem/métodos , Adenina/sangre , Calibración , Desoxicitidina/sangre , Combinación de Medicamentos , Estabilidad de Medicamentos , Emtricitabina , Humanos , Guías de Práctica Clínica como Asunto/normas , Control de Calidad , Estándares de Referencia , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Comprimidos/análisis , Tenofovir , Equivalencia Terapéutica , Factores de TiempoRESUMEN
A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the simultaneous estimation of hydrochlorothiazide, quinapril and its metabolite quinaprilat in human plasma. After solid phase extraction (SPE), the analytes and IS were chromatographed on a hypurity C8 (100 mm x 2.1 mm i.d., 5 microm particle size) column using 2 microL injection volume with a run time of 2.8 min. An isocratic mobile phase consisting of 0.5% (v/v) formic acid:acetonitrile (25:75, v/v) was used to separate all these drugs. The precursor and product ions of these drugs were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring mode (MRM) without polarity switch. The proposed method was validated over the range of 5-500 ng/mL for hydrochlorothiazide method and 5-1500 ng/mL for quinapril and quinaprilat. Inter-batch and intra-batch precision (coefficient of variation - % CV) across five validation runs lower limit of quantitation (LLOQ), lower quality control (LQC), middle quality control (MQC), higher quality control (HQC) and upper limit of quantitation (ULOQ) was less than 15. The accuracy determined at these levels was within +/-13% in terms of relative percentage error.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Hidroclorotiazida/sangre , Espectrometría de Masas en Tándem/métodos , Tetrahidroisoquinolinas/sangre , Humanos , Hidroclorotiazida/farmacocinética , Quinapril , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Tetrahidroisoquinolinas/farmacocinética , Equivalencia TerapéuticaRESUMEN
A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the estimation of clonidine in human plasma. Clonidine was extracted from human plasma by using solid-phase extraction technique. Nizatidine was used as the internal standard. A Hypurity C18 (50 mm x 4.6 mm i.d., 5 microm particle size) column provided chromatographic separation of analyte followed by detection with mass spectrometry. The method involves a rapid solid-phase extraction from plasma, simple isocratic chromatography conditions and mass spectrometric detection that enables detection up to picogram levels with a total run time of 3.0 min only. The method was validated over the range of 50-2500 pg/mL. The absolute recoveries for clonidine (71.86%) and IS (69.44%) achieved from spiked plasma samples were consistent and reproducible.
