Abstract
A simple, precise, isocratic, reverse phase high performance liquid chromatography (HPLC) method was developed for the simultaneous determination of curcuminoids: viz. curcumin(C), desmethoxycurcumin (DMC), bisdesmethoxycurcumin (BDMC) and celecoxib at single wavelength using a Agilent RP C18, 4.6 mm × 150 mm, 5 μm XDB column. The run time was 18 min. The influence of mobile phase composition, injection volume, mobile phase pH, flow rate, temperature, and detector wavelength on resolution was investigated. The method was validated with respect to precision, accuracy, and linearity. The LOD and LOQ were found to be 0.3 and 1 μg/mL, respectively for curcumin and 0.03 and 0.1 μg/mL for respectively for celecoxib. Linearity range was from 1- 20 μg/ mL for curcumin and from 0.1-2 μg/mL for celecoxib. Further, the proposed method was found to be reproducible and convenient for stability-indicating analysis of curcumin–celecoxib pH sensitive nanoparticles.
Highlights
It is a well known fact curcumin; a coloring principle in turmeric has emerged as the most exploited phyto-constituents for its wide spectrum of biological activities
Methanol was replaced by acetonitrile and used in various ratios, with the result that splitting peaks were observed for both drugs. Both the drugs were tried with combination of methanol and water (1% acetic acid; adjusted to pH of 3.0 using 50% triethanolamine) at various ratios, still curcumin was unable to separate into three peaks
In the chromatogram of curcumin-celecoxib samples extracted from pH sensitive nanoparticles, separate peaks were observed corresponding to the three curcuminoids and celecoxib
Summary
It is a well known fact curcumin; a coloring principle in turmeric has emerged as the most exploited phyto-constituents for its wide spectrum of biological activities. Celecoxib was the first specific inhibitor of a cycloxygenase-2 (COX-2) with little or no effect on the gastrointestinal (GI) tract and kidney It has been recommended for the treatment of rheumatoid arthritis, and osteoarthritis for its anti-inflammatory activity. The synergistic use of these drugs could augment the anti-inflammatory activity Been demonstrated in osteoarthritis synovial adherent cells wherein curcumin synergistically enhances the growth inhibitory and proapoptotic effects of celecoxib [7] Considering these benefits, it is worthwhile to explore the combination of both these drugs in treatment of inflammatory conditions. 1 mL of methanolic stock solution of curcumin and 0.2 mL of methanolic stock solution of celecoxib were taken in 10 mL amber volumetric flask and heated for 1 hour in water bath maintained at 80oC to study the heat degradation. The resultant cooled mixture was diluted to 10 mL with methanol, injected in triplicate and the chromatogram was run as described previously
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