Development and validation of a column high-performance liquid chromatography method for quantification of ganaphaliin A and B in inflorescences of Gnaphalium liebmannii Sch. Bp ex Klatt

An HPLC method was developed for the simultaneous determination of gnaphaliin A and B, active compounds of Gnaphalium liebmannii Sch. Bp ex Klatt. The HPLC separation was performed on an Inertsil ODS-3 (150 x 4.6 mm id, 5 microm) RP C18 column operated at 40 degrees C; the isocratic mobile phase was 0.02% aqueous orthophosphoric acid-methanol-acetonitrile (50 + 30 + 20, v/v/v), with a run time of 20 min and flow rate of 1.5 mL/min. Detection with a photodiode array detector (PDAD) was at 270 nm. The method was validated for linearity, repeatability, LOD, and LOQ. The LOD and LOQ for gnaphaliin A and B were found to be in the range of 0.4-0.5 and 1.0-1.4 microg/mL, respectively. This is the first report of an analytical method developed for the quantitative analysis of flavones from Gnaphalium species by HPLC-PDAD with applications for raw material and commercial products.