DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS DETERMINATION OF CLOPAMIDE, DIHYDROERGOCRISTINE MESYLATE AND RESERPINE IN TABLETS
Keywords:RP-HPLC, Method development, Validation, Clopamide, Dihydroergocristine mesylate, Reserpine
Objective: To develop a new, rapid, precise and sensitive RP-HPLC method for the simultaneous determination of Clopamide (CPM), Dihydroergocristine mesylate (DHECM) and Reserpine (RSP) in tablets.
Methods: Caffeine (CAFF) was used as internal standard, a BDS Hypersil column C8 (4.6 x 150 mm) with mobile phase containing a mixture of inorganic phase (ammonium acetate solution in water) and organic phase (acetonitrile and methanol), adjusted with phosphoric acid (10% v/v) to a pH-3.5, the flow rate was 2.0 ml/min, UV detection was performed at a wavelength of 220 nm and injection volume was 20 Î¼l. The validation of this method was done as per ICH guidelines.
Results: Retention times were observed as 1.277, 2.397, 3.799 and 5.202 min for Caffeine, Clopamide, Dihydroergocristine mesylate and Reserpine, respectively. Linearity ranges were observed to be: 1.50-300.00, 2.50-500.00 and 1.00-200.00 Î¼g/ml for Clopamide, Dihydroergocristine mesylate and Reserpine respectively. Relative Standard Deviation did not exceed 2.00%. Detection limits were observed to be 0.08, 0.07 and 0.05 Î¼g/ml for Clopamide, Dihydroergocristine mesylate and Reserpine, respectively.
Conclusion: This method was applied, the first time as simple, rapid, precise and sensitive method for determination of Clopamide, Dihydroergocristine mesylate and Reserpine either alone or in combination, in pure form and in pharmaceutical preparations without interference with any excipient.
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