Estimation of rosuvastatin in human plasma by HLPC tandem mass spectroscopic method and its application to bioequivalence study

Abstract

Um método empregando LC-MS/MS foi desenvolvido para a análise de rosuvastatina em plasma humano, usando atorvastatina como padrão interno. Rosuvastatina é um fármaco para redução de lipídeos e prescrita para o tratamento do hipercolesterolemia e de dislipidimia. A extração em fase sólida (SPE) foi usada para purificação e pré-concentração do analito a partir da matriz do plasma humano. A separação cromatográfica foi conseguida em 6.0 min empregando fase móvel composta de 0.2% de ácido fórmico em água e acetonitrila (40: 60,v/v, na vazão de 1.0 mL min -1 e coluna YMC J'sphere ODS H-80, 150 x 4.6 mm, partículas de 4.0 μm. Na saída da coluna, a fase móvel foi dividida, sendo que 200 μL foram dirigidos para o espectrômetro de massas e 800 μL para o descarte. Pelo Monitoramento de Reação Múltipla (MRM), as transições foram medidas no modo positivo em m/z 482 -258 para rosuvastatina e m/z 559 -440 para o padrão interno, respectivamente. Uma validação detalhada do método foi realizada seguindo as recomendações do FDA americano e as curvas analíticas foram lineares no intervalo de 1.00 ng mL -1 a 50.00 ng mL -1 com coeficiente de correlação médio maior que 0.99. A recuperação absoluta foi maior que 50.14% para rosuvastatina e 54.65% para o padrão interno. Rosuvastatina foi estável por 138 dias a -70 ± 5 °C e por 24 horas à temperatura ambiente. Após a extração do plasma, as amostras reconstituídas de rosuvastatina permaneceram estáveis no auto injetor, a 10 °C, por 8 horas. Depois de submetidas a três ciclos de congelamento/descongelamento, não houve mudanças na recuperação do analito. O método é simples, específico, sensível, preciso, exato e apropriado para aplicações em bioequivalência e estudos farmacocinéticos. Foi aplicado com sucesso em um estudo piloto de bioequivalência da rosuvastatina, comprimidos -Zydus, Cadila, India versus comprimidos -Crestor, Astra Zeneca, EUA, em voluntários sadios do sexo masculino.A LC-MS/MS method has been developed for the estimation of rosuvastatin in human plasma using atorvastatin as internal standard. Rosuvastatin is a lipid-lowering drug prescribed for the treatment of hyper-cholestrolemia and dyslipidimia. Solid phase extraction (SPE) was used for the purification and pre-concentration of analyte from human plasma matrix. The chromatographic separation was achieved within 6.0 min by an isocratic mobile phase containing 0.2% formic acid in water and acetonitrile (40: 60, v/v), flowing through YMC J' Sphere ODS H-80, 150 x 4.6 mm, 4.0 μm analytical column, at a flow rate of 1.0 mL min -1 with split of 200 μL to mass spectrometer and 800 μL to waste. Multiple reaction monitoring (MRM) transitions were measured in the positive mode at m/z 482 and 258 for rosuvastatin and m/z 559 and 440 for internal standard respectively. A detailed validation of the method was performed as per USFDA guidelines and the standard curves were found to be linear in the range 1.0 ng mL -1 to 50.0 ng mL -1 with the mean correlation coefficient more than 0.99. The absolute recovery was more than 50.14% f...

Quantitative determination of rosuvastatin in human plasma by ion pair liquid–liquid extraction using liquid chromatography with electrospray ionization tandem mass spectrometry

“…Hull et al reported an automated solid phase extraction [14] which was successfully applied to a series of clinical trials on bioavailability, pharmacokinetic and metabolic [15] study of rosuvastatin. A similar method employing solid phase extraction and HPLC-MS/MS with a relative higher LLOQ of 1 ng mL −1 was recently reported by Singh et al [16]. Recently, a microbore HPLC in combination with tandem MS which was cross-validated to provide similar information on the concentration of rosuvastatin exhibited an ability of greatly reduced sample consumption [17].…”

Development and Validation of Stability-Indicating HPLC Methods for Quantitative Determination of Pravastatin, Fluvastatin, Atorvastatin, and Rosuvastatin in Pharmaceuticals

“…Several methods based on different techniques have been reported for the analysis of PS, FVS, ATC, and RC in biological fluids. These include high-performance liquid chromatography-mass spectrometry (HPLC=MS) (Gao et al 2007;Lan et al 2007;Borek-Dohalsky et al 2006;Di Pietro et al 2006;Nirogi et al 2006;Oudhoff et al 2006;Xu et al 2006;Hermann, Christensen, and Reubsaet 2005;Singh et al 2005;Trivedi 2005;Zhu and Neirinck 2003) and HPLC (Bahrami et al 2005;Kumar et al 2006;Vittal et al 2006;Um et al 2005;Bauer et al 2005;Zarghi et al 2005;Altuntas and Erk 2004;Al-Rawithi, Hussein, and Alzahrani 2003). Although these methods are sensitive enough to permit their use in determination of PS, FV, ATC, and RC in urine, plasma, and serum, few methods are reported for assay of statins in pharmaceuticals.…”

Determination of Rosuvastatin Calcium in bulk and Pharmaceutical dosage forms by using UV-Spectrophotometric method

“…Common dosage forms are tablets and capsule (Ahmad et al, 2012). As per literature survey it reveals that, only few analytical techniques have been developed for the estimation of rosuvastatin calcium in bulk and pharmaceuticals such as High performance thin layer chromatography (HPTLC) method (Uma Devi et al, 2011), one Reverse phase high performance liquid chromatographic (RP-HPLC) method (Pandya et al, 2010) and few LC-MS methods (Singh et al, 2005). In this study an attempt was made to develop a simple spectrophotometric method for the estimation of the present drug in dosage form.…”