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Abstract

BIO ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF LERCANIDIPINE AND ATENOLOL IN HUMAN PLASMA BY USING RP-HPLC

Gokul P. and Ravichandran S.*

ABSTRACT

A simple, sensitive, precise and accurate high performance liquid chromatographic (HPLC) method was developed and validated for the estimation of Lercanidipine and Atenolol in human plasma. The chromatographic separation was achieved with the reverse phase column Phenomenox Gemini C18 (150mmX4.6, 5μ) and the mobile phase consisted of acetonitrile and 10mM ammonium acetate buffer (80:20 v/v) at ph-3.2(adjusted with acetic acid) at a flow rate of 0.2mL/min. Haloperidol was used as an internal standard. The wavelength used for the detection of Lercanidipine and Atenolol was 275nm with total run time of 10 minutes. The retention times of Lercanidipine, Atenolol and Haloperidol was found to be 6.6, 3.5 and 4.3 minutes respectively. The calibration curve was linear (r2> or =0.99) ranging from 5 to 2000ng/mL for Lercanidipine and 10 to 3000ng/mL for Atenolol. The lower limit of quantification was 15ng/mL for Lercanidipine and 30ng/mL for Atenolol. Interday precision were lower than 5% (CV) and accuracy ranged from 98 to 102 % for Lercanidipine and 96 to 102% for Atenolol in terms of percent accuracy. Mean extraction recovery was found to be 75 to 80% for both Lercanidipine and Atenolol respectively. The method was successfully developed and validated in human plasma for excellent selectivity, accuracy, precision, recovery and stability in compliance to international regulatory guidelines.

Keywords: HPLC, Lercanidipine, Atenolol, internal standard, human plasma, validation.


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