Chromatographic Estimation of Curcumin and Parthenolide Using a Validated RP-HPLC Method

A Reverse-Phase High-Performance Liquid Chromatographic (RPHPLC) method was developed to estimate curcumin and parthenolide simultaneously.

Method: A Waters HPLC (HPLC/2487) with a UV/Vis detector was used for the analysis. Chromatographic separation was carried out using a reverse-phase C18 column (250 mm × 4.6 mm, 5 µm particle size). A mobile phase consisting of acetonitrile and 0.1% orthophosphoric acid in water was pumped at a flow rate of 1 mL/min. Quantification of curcumin and parthenolide was performed using a UV/Vis detector set at 222 nm, identified as the isosbestic point for both analytes. The developed method was evaluated for specificity, linearity, accuracy, precision, robustness, LOD, and LOQ in accordance with the ICH Q2 (R2) guidelines established by the International Council for Harmonisation.

Results: Curcumin had a linearity and range of 2-10 µg/mL, while parthenolide had a range of 5-25 µg/mL. Curcumin's R2 was 0.9992, while parthenolide's was 0.9899. The developed method exhibited a Limit of Quantification (LOQ) of 5.09 µg/mL for parthenolide and 1.68 µg/mL for curcumin. The corresponding Limits of Detection (LOD) were determined to be 1.01 µg/mL and 3.06 µg/mL, respectively. Furthermore, the method demonstrated acceptable levels of precision and accuracy.

Discussion: The validated RP-HPLC method successfully quantified both curcumin and parthenolide together, demonstrating the linearity, accuracy, and precision within the limits. Low values of LOD and LOQ confirmed the sensitivity of this method for the quantification of even small amounts of these drugs.

Conclusion: The method demonstrated practical application for analysing a combination of these drugs in various pharmaceutical dosage forms. Curcumin and parthenolide can be accurately and simultaneously quantified using the approved RP-HPLC method, which has better sensitivity, a shorter runtime, and less spectrum interference at 222 nm. It is ideal for routine quality control, stability investigations, and standardisation of two-component phytopharmaceutical formulations because of its compliance with regulatory criteria and flexibility to complicated matrices.