Determination of parthenin in Parthenium hysterophorus L. by means of HPLC-UV: Method development and validation

Abstract

Parthenium hysterophorus L. or Santa Maria feverfew is a plant used in Cuba since antiquity for the treatment of several diseases. Nowadays it is still used as an antipyretic and antiparasitic agent. Parthenin, a sesquiterpene lactone, is the active secondary metabolite and the major component of the plant. In this study the development and validation of a HPLC method for the determination of parthenin in the powdered plant material are presented, making it possible to perform quality control on preparations containing P. hysterophorus. Firstly, parthenin was isolated from the plant material in order to use this as reference material. During the method development, the extraction procedure, sample preparation, and HPLC conditions were evaluated and optimized. The final method was fully validated in terms of calibration model, precision, accuracy, and specificity. Based on these results, it was concluded that the developed HPLC method is suitable for the determination of parthenin using a single-point calibration. The calibration model was linear in the concentration range from 0.05 to 0.25 mg/ml. Analysis on different days showed that the method was precise with an average concentration of 4.73% and RSD of 1.39%. A recovery experiment was performed resulting in a 95% confidence interval between 97.5% and 100.4%, meaning that the method is also accurate. Specificity was confirmed by the investigation of the peak purity. Using this newly validated method the quality of the plant material of P. hysterophorus, used as an active principle in pharmaceutical preparations, can be guaranteed.