HPLC Analysis of Oxindole Alkaloids in Uncaria Tomentosa : Sample Preparation and Analysis Optimisation by Factorial Design

Introduction Uncaria tomentosa (“cat's claw”) is widely used for the treatment of some infectious and inflammatory diseases. Oxindole alkaloids are regarded as the most important components responsible for the biological activities attributed to the plant. Their analysis require efficient sample preparation and suitable reference standards but few are commercially available. Objective To develop and validate a HPLC analytical method for oxindole alkaloids in Uncaria tomentosa with emphasis on sample preparation. Method Factorial experimental designs were used for the optimisation of both sample preparation and chromatographic separation. The optimised sample preparation involved extraction with aqueous ethanol, and the granulometry of the powdered plant material significantly influenced extraction yields. Mitraphylline was used as a calibration reference for the determination of total alkaloids. Results The method was fully validated and showed good selectivity, linearity (r2 ≥ 0.9996), accuracy (≥ 96%) and precision (RSD