OPTIMIZATION AND VALIDATION OF HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD WITH UV DETECTION FOR THE DETERMINATION OF EPROSARTAN MESYLATE IN RAT PLASMA AND URINE

Randa Salam, Ghada Hadad, Eman Hameed
unpublished
Objectives: The objectives of this research were to develop and validate a reverse phase-high performance liquid chromatography (RP-HPLC) method for determination of eprosartsan (ES) in rat plasma and urine using diclofenac sodium (DC) as an internal standard (I.S). Methods: Plasma sample was treated with deproteinizing agent which is acetonitrile. The proposed method was based on using a 250 x 4.6 mm (i.d.) Phenomenex cyanopropyl column (5 µm particle size) with mobile phase consisting of 25
more » ... consisting of 25 mM ammonium acetate buffer pH 3.7-acetonitrile, in ratio of (70:30, v/v) and UV detection at 240 nm with flow rate of 1.2 mL min-1. Results: ES eluted at 6.09 ± 0.07 minutes while the elution time for DC (I.S) was 10.69 ± 0.08 min. The linear calibration range for ES was observed between 0.05-20 μgmL-1 in rat plasma and urine. The method was found to be accurate with 99.25% and 99.70% recovery in rat plasma and urine respectively, precise as the coefficient of variation (CV%) of inter day and intraday precision was less than 2%.The limit of detection (LOD) was found to be 3.29 x10-2 μg mL-1 and 3.56 x10-2 μgmL-1 in rat plasma and urine respectively, while the limit of quantification (LOQ) was found to be 1.09x10-1 μg mL-1 and 1.19x10-1 μg mL-1 in rat plasma and urine respectively. Conclusion: A novel specific, accurate, precise chromatographic method was developed for quantitative estimation of ES in rat plasma and urine. The validation study of the proposed method was successfully carried out and the method was found suitable and economic for routine determination of eprosartan in rat plasma and urine.
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