Fast analysis of β-ecdysone in Brazilian ginseng (Pfaffia glomerata) extracts by high-performance liquid chromatography using a fused-core column

摘要

The recent development of fused-core technology in HPLC columns is enabling faster and highly efficient separations. This technology was evaluated for the development of a fast analysis method for ?2-ecdysone in extracts of Pfaffia glomerata. A step-by-step strategy was used to optimize temperature (30a€“55 ?°C), flow rate (1.0a€“2.0 mL mina?’1), mobile phase composition (mixtures of water and methanol or acetonitrile) and equilibration time (1a€“5 min). A gradient method has been developed using two solvents: 0.1% acetic acid in water and 0.1% acetic acid in acetonitrile. Optimized conditions provided a method for the separation of ?2-ecdysone in approximately 2 min with a total analysis time (sample-to-sample) of 9 min, including the return to initial conditions and the re-equilibration of the column. Evaluation of chromatographic performance revealed excellent intraday and interday reproducibility (99.5%), resolution (2.78), selectivity (1.13), and peak symmetry (1.09) while presenting low limits of detection (0.20 mg La?’1) and quantitation (0.67 mg La?’1). The robustness of the method has also been calculated according to the concentration/dilution of the sample. Several sample solvents were evaluated and the best chromatographic results were obtained using 80% methanol in water. Finally, the developed method was validated with different extracts of Pfaffia glomerata samples.