METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATION OF CURCUMIN AND PIPERINE BY RP-HPLC
Keywords:Method development, Method validation, RP-HPLC, Curcumin, Piperine
Objective: To develop a simple, sensitive, specific, accurate reverse phase high performance liquid chromatographic (RP-HPLC) method for the estimation of curcumin and piperine.
Methods: The separation was done using a column Inertsilâ€“ODS C18 (250 mm Ã— 4.6 mm, 5Âµ particle size) and mobile phase composed of methanol: water (45:55 v/v), flow rate at 1 ml/min and detection was carried out at 282 nm with photodiode array (PDA) detector.
Results: The separation of curcumin and piperine were found to be at the retention time of 2.433 min and 3.095 min, respectively. The method was found to be linear at a concentration range 20-80 Âµg/ml for curcumin and piperine. The limit of detection (LOD) and limit of quantification (LOQ) was found to be 0.05Âµg/ml and 0.17Âµg/ml for curcumin and 0.18Âµg/ml and 0.53Âµg/ml for piperine respectively. The average percentage recoveries of curcumin and piperine were in the range of 98-100.6%.
Conclusion: A simple and sensitive reverse phase high performance liquid chromatographic method was developed for the estimation of curcumin and piperine.
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