DEVELOPMENT AND VALIDATION OF BIO-ANALYTICAL RP-ULTRA FAST LIQUID CHROMATOGRAPHIC METHOD FOR SIMULTANEOUS ESTIMATION OF CLOPIDOGREL AND ROSUVASTATIN IN HUMAN PLASMA

ABSTRACT: This manuscript describes a simple, sensitive, rapid and precise Ultra fast liquid chromatographic (UFLC) method was developed and validated for the simultaneous determination Clopidogrel and Rosuvastatin in human plasma according to USFDA draft guidelines. In the current study, the analysis was performed on phenomenex C8 (250 × 4.6mm, 5μm) column using phosphate buffer (pH-2.5) and acetonitrile (45: 55 v/v) as mobile phase at flow rate of 1.2 mL/min. In this developed method Clopidogrel and Rosuvastatin eluted at a retention time of 4.697 and 3.337 min respectively. The proposed method is having linearity in the concentration range from 10 to 50μg/mL of Clopidogrel and Rosuvastatin. The current method was validated with respect to linearity; precision, lowest limit of detection (LOD), accuracy and recovery according to the USFDA guidelines. The system consisted of a pump (Shimadzu, prominence, UFLC), with 20µl sample injector, along with a PDA detector at a wavelength of 243 nm and 220 nm for Rosuvastatin and Clopidogrel respectively. Data was compiled using Shimadzu LC Solution software. A good linear relationship over the concentration range of 10-50µg/ml was shown. Validation of the method was carried out as per the USFDA draft guidelines. The method developed was found to be precise, accurate, specific, linear and selective. Statistical analysis shows that the method is reproducible and selective for the estimation of Clopidogrel and Rosuvastatin in dosage form