Simultaneous high-performance liquid chromatographic determination of a rhein-diclofenac prodrug and its active compounds.

In order to study the hydrolytic characterization of an anti-inflammatory prodrug (RD-1) in vitro, a simple, specific, precise and accurate method for the simultaneous determination of prodrug and its two hydrolytic active compounds was development using reverse phase high-performance liquid chromatography (RP-HPLC). The chromatographic separation was performed on an ODS-2 C18 column (250 mm x 4.6 mm, 5.0 microm particle size) with a simple elution programme. The mobile phase was methanol-0.1%phosphoric acid solution (adjusted pH to 2.3) (80:20, v/v); wavelength of 257nm and mobile phase flow rate of 1.0 mL/min was utilized for the quantitative analysis. Excellent linear behaviors over the investigated concentration ranges were observed with the values of R2 higher than 0.999 for all the analytes. The validated method was successfully applied to the simultaneous determination of prodrug and its active components.