Simultaneous SPE-LC determination of three flavonoid glycosides of naringin, neohesperidin and hesperidin in Da-Cheng-Qi decoction

In this paper a rapid solid-phase extraction (SPE) high-performance liquid chromatographic (HPLC) method coupled with ultraviolet detector (UV) has been developed and validated for the simultaneous analysis of three flavonoid glycosides of naringin, hesperidin and neohesperidin in Da-Cheng-Qi (DCQ) decoction, a widely used purgative prescription of traditional Chinese medicine (TCM). The chromatographic separation was performed on an Agilent Zorbax SB-C-18 (5 mu m, 250 x 4.6 mm) analytical column by isocratic elution using a mixture of methanol: water (0.2% formic acid), 60:40 (v/v) at 45 degrees C. This method was validated in terms of recovery, linearity, accuracy, precision and repeatability. The recoveries from spiked control samples were 94.3, 92.6 and 95.7% for naringin, hesperidin and neohesperidin, respectively with RSD values less than 5.0%. The standard calibration curves were linear between 3.9 and 125 mu g mL(-1) for hesperidin, 15.6-500 mu g mL(-1) for neohesperidin and naringin with regression coefficient r(2) > 0.9995. The intra-day and inter-day precisions of this method were evaluated with the RSD values less than 2.0% for all compounds. This method was successfully used to determine the three target compounds in 10 batches of Da-Cheng-Qi decoction.