Authors: G. J. Yang 1 and L. Lv 2
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  • 1 School of Pharmacy, China Pharmaceutical University, Nanjing, 210009, P. R. China
  • | 2 College of Chemistry and Chemical Engineering, Yangzhou University, Yangzhou, 225002, P. R. China
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Summary

A simple and sensitive method of high-performance liquid chromatography with fluorescence detection (HPLC-FLD) was developed for the determination of icariin in capsules by precolumn chelation with aluminum. In order to obtain a stable fluorescence signal, the reaction conditions of the fluorescent chelation complex between icariin and aluminum were investigated in detail. Chromatography was carried out on an Agilent Zorbax Extend C18 column (150 mm × 4.6 mm, 5.0 μm) using methanol as mobile phase at a flow rate of 1.0 mL min−1. The excitation and emission wavelengths were set at 430 and 480 nm, respectively. At optimum conditions, the calibration curve was linear in the concentration range from 0.010 to 100.0 μg mL−1 with the limit of detection of 3.5 ng mL−1 (S/N = 3). A comprehensive method was validated for precision and accuracy. The method described here has been successfully applied for the determination of the icariin content in a capsule with satisfactory results.

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