Abstract

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING RP-HPLC METHODS FOR DETERMINATION OF IMPURITIES IN RILPIVIRINE HYDROCHLORIDE CAPSULES

Nuzhath Fathima*, P. Raveendra Reddy, M. Brahmaiah, Qasimullah, G. Abdul Huq

ABSTRACT

A rapid, reverse phase HPLC method has been developed for the determination of Rilpivirine Hydrochloride and its related impurities. These impurities were isolated from crude sample of Rilpivirine Hydrochloride using reverse phase HPLC. The IUPAC names of impurities are Impurity-A is 4-(4-Hydroxypyrimidin-2-ylamino) benzonitrile, Impurity-B is (E)-3-(4-(2-(4-Cyanophenylamino) pyrimidin-4-ylamino)-3, 5-dimethylphenyl) acrylamide, Impurity-C is 4-[(4-Chloro-2-pyrimidinyl) amino] benzonitrile, Impurity-D is 4-{[4- ({4-[(Z)-2-Cyanovinayl]-2,6-dimethylphenyl} amino) pyrimidin-2-yl] amino} benzonitrile, Impurity-E is 4-[[4-[[4-[(E)-2-cyanoethenyl]-2, 6- dimethylphenyl] amino-2-pyrimidinyl] amino] benzonitrile monohydrochloride and Impurity- F is (2E)-3-(4-Amino-3,5-dimethylphenyl) acrylonitrile hydrochloride. The separation method was optimized in Gemini phenomenex (250x4.6) m, 5μ column with buffer of 15 mm KH2PO4 and pH adjusted to 3.0 with H3PO4 and the organic modifier was methanol. The gradient elution programme is 0.01/30, 35/85, 40/85, 41/30, 45/stop and flow rate is 1.20 mL/min. The retention times of Rilpivirine Hydrochloride and its impurities are 1.00, 11.90, 14.93, 19.19, 22.44, 23.53 and 27.12 respectively. The developed method was validated in terms of system suitability, specificity, linearity range, precision, accuracy, limits of detection and quantification for the impurities following the ICH guidelines. Therefore, the proposed method is suitable for the simultaneous determination of Rilpivirine Hydrochloride and its related impurities.

Keywords: Rilpivirine Hydrochloride, Impurities, RP-HPLC, Gemini phenomenex C18, Inertsil ODS, Unison, ZodiacSIL AQ-C18 and validation.