A validated stability-indicating HPLC method for analysis of glabridin prodrugs in hydrolysis studies.

A simple, selective and precise stabilityindicating HPLC method for determination of glabridin diacetate and dihexanoate prodrugs was developed, validated and applied to the enzymatic and chemical hydrolysis studies. The chromatographic separation was achieved on a reverse phase C18 (Thermo Hypersil-Keystone, 250 × 4.6 mm, 5 micron) column using the mixture of acetonitrile and water as mobile phase. Elution of the mobile phase was operated on isocratic (acetonitrile 76%: water 24%) for 9 min, followed by gradient (acetonitrile from 76% to 90%) within 9 min and isocratic (acetonitrile 90%: water 10%) for 12 min at 1 mL/min flow rate, detected at 280 nm. The method was validated for specificity, accuracy, precision, linearity and limit of quantitation following the International Conference on Harmonization (ICH) guidelines. The method is effective for the separation of glabridin diacetate and glabridin dihexanoate from glabridin, its parent drug and successfully used in these prodrugs hydrolysis studies.