Abstract
A flow injection (FI) micro-column system coupled with high performance liquid chromatography (HPLC) was proposed for the pre-separation and determination of active organic component (ecdysterone) in traditional Chinese medicine, Loulu. The factors influencing separation performance were investigated and optimized. Under the optimal conditions, the contents of ecdysterone in Loulu were determined by HPLC system using MeOH-H2O (40∶60,V/V) as the mobile phase at a flow rate of 1.0 mL/min. The calibration curve was linear in the range of 0.5∼100 mg/L of ecdysterone concentrations. The detection limit of the analyte was 0.11 μmol/L (3σ) with a precision of 0.38% RSD (n=7,c=10.0 mg/L). The average recovery of the method was 98.7%. The proposed method has been applied to determine ecdysterone in practical samples, and the determined values by both external standard method and standard addition method were in good agreement. Compared to the traditional solid extraction method, the system proposed has the advantages of simple procedure, good reproducibility, minimum volume requirement, reduction of matrix interference and low contamination risk.
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