DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF DICLOFENAC DIETHYLAMINE AND CURCUMIN IN TRANSDERMAL GELS

Abstract

A reversed phase high performance liquid chromatographic (RP-HPLC) method using Symmetry C18, 5.0-µm column was developed for simultaneous determination of Diclofenac diethylamine (DDEA) and Curcumin (CRM). Mobile phase consisted of orthophosphoric acid 0.1% and acetonitrile (65:35), and the flow rate was 1.5 mL/min and elution was monitored initially at 425 nm for CRM, after 14.7 min wavelength was changed to 275 nm for detection of DDEA.

Response was a linear function of concentration over the range 0.245–1.96 µg/mL (R² = 0.9998) for DDEA and 55–445 ng/mL (R² = 0.9992) for CRM and the limits of detection were 245.5 ng/mL for DDEA and 55.30 ng/mL for CRM. The limit of quantitation was 491.1 ng mL − 1 for DDEA and 165.87 ng mL − 1 for CRM. The method was validated in accordance with ICH guidelines. The method was used for simultaneous quantification of DDEA and CRM in transdermal gel formulations and to check content uniformity. The excipients present in the formulation did not interfere with the assay. The method is suitable for application in quality-control laboratories, because it is simple and rapid with good accuracy and precision.

Keywords:

• curcumin
• diclofenac diethylamine
• method validation
• RP-HPLC
• simultaneous quantification
• UV-detector

ACKNOWLEDGMENTS

Hema Chaudhary thanks the University Grants Commission, Government of India for providing financial support for the UGC Major Research Project (F. No: 32-136/2006(SR)). The authors are also grateful to Arbro Labs, Ltd., New Delhi, India, for providing the drug sample of CRM and providing the facility for completion of the work.

Notes

(Y)* = a +bX where Y is the area and x concentration in ng mL⁻¹.

^a data represents mean ± SD; n = 3.