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 C-18 column (250 mm x 4.6 mm, 5.0 mu m 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 257 nm 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 R-2 higher than 0.999 for all the analytes. The validated method was successfully applied to the simultaneous determination of prodrug and its active components.