Quality by design approach for enantiomeric evaluation by RP-HPLC method of Rivaroxaban and its chiral impurity

Rivaroxaban is an anticoagulant that presents as a mechanism of action the direct and selective inhibition of activated coagulation factor X (FXa). This molecule has a chiral center but only the enantiomer (S)-Rivaroxaban presents pharmacological activity being (R)-Rivaroxaban an impurity. Quality by Design (QbD) is a fundamental model of pharmaceutical quality used in the development of products and processes. The QbD approach allows the identification of critical parameters to predefined quality attributes and is conducted by risk analysis and design of experiments (DoE) promoting a broader knowledge about the analytical method which is relevant for methods of quantification of impurities. The chiral analysis presents as a challenge the separation of compounds with identical physical-chemical properties. Thus, it is fundamental the use of specific techniques to resolve enantiomers, demonstrating the need for the development of appropriate and reliable methods for this purpose. This work aimed to develop and validate an analytical method capable of separate and quantify S-Rivaroxaban (S-RIV) and its enantiomeric impurity R-Rivaroxaban (R-RIV). For that goal, it was used the QbD approach for the development of a suitable and reproductive method using high-performance liquid chromatography (HPLC). The chromatographic conditions were a Chiralpak & REG; AD-RH (150 x 4.6 mm; 5 & mu;m) column, mobile phase composed of acetonitrile (ACN): water pH 4.5 (92:8 v/v) with a flow of 0.35 mL/min, detection at 250 nm, a temperature of 40 degrees C and total analysis time of 12 min. The analytical method was validated following the official guidelines, demonstrating selectivity, linearity, precision, accuracy and sensitivity presenting a limit of quantification (LOQ) of 0.68 & mu;g/mL for S-RIV and 1.0 & mu;g/mL for the chiral impurity. After the development and validation steps, the presence of R-RIV impurity in marketed tablets was performed using the proposed method. This study resulted on the development and validation of a reliable and appropriate chiral analytical method for its aim bringing a valuable contribution to quality control routines.