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Abstract

VALIDATED BIO-ANALYTICAL METHOD DEVELOPMENT FOR SIMULTANEOUS ESTIMATION OF CLOPIDOGREL AND ASPIRIN IN HUMAN PLASMA BY RP-ULTRA FAST LIQUID CHROMATOGRAPHY

Jinesh Bahubali Nagavi*, Bannimath Gurupadayya, Preethi G. A

ABSTRACT

This manuscript describes a simple, sensitive, rapid and precise Ultra fast liquid chromatographic (UFLC) method for the simultaneous determination Clopidogrel and Aspirin 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 (35: 65 v/v) as mobile phase at flow rate of 1.5 mL/min. In this developed method Clopidogrel and Aspirin eluted at a retention time of 2.697 and 5.337 min respectively. The proposed method is having linearity in the concentration range from 10 to 50μg/mL of Clopidogrel and Aspirin. 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 230 nm and 252 nm for Aspirin 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 Aspirin in dosage form. Keywords:

Keywords: Bioanalytical, Clopidogrel, Aspirin, RP-UFLC, USFDA


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