Desenvolvimento e validação de método por cromatografia líquida em fase reversa para análise de rivaroxabana

Abstract

Rivaroxaban is an oral anticoagulant drug indicated for deep venous thromboembolism prophylaxis. A reversed-phase liquid chromatography (RP-LC) method was developed and validated for the determination of rivaroxaban in pharmaceutical dosage forms. The LC method was carried out on a Phenomenex Fusion-RP C18 column (150 mm x 4.6 mm i.d.), maintained at 40 °C. The mobile phase consisted of ultrapure water–acetonitrile (30:70, v/v), run at a flow-rate of 0.7 mL/min and using photodiode array (PDA) detection at 249 nm. The chromatographic separation was obtained with retention time of 2.9 min, and was linear over the range of 0.04 – 200 μg/mL (R2 = 0.9992). The specificity and stability-indicating capability of the method was through degradation studies, and showing also, that there was no interference of the excipients and degradation products in the quantification of the drug. The accuracy was 99.77% with bias lower than 1.67%. The limits of detection and quantitation were 0.012 and 0.041 μg/mL, respectively. The procedure was validated evaluating parameters such as the specificity, linearity, precision, accuracy, limits of detection and quantitation, robustness and system suitability test, given results with acceptable range. In parallel, the anti-factor Xa assay was performed for the activity evaluation of rivaroxaban in relation to sodium enoxaparin. Moreover, the in vitro cytotoxicity test of the degraded products showed significant differences (p < 0,05). The proposed method was applied for dissolution studies, analysis in human plasma and the analysis of rivaroxaban in tablet dosage forms, contributing to assure the safety and therapeutic efficacy.