Simultaneous determination of ten bioactive compounds from the roots of Cynanchum paniculatum by using high performance liquid chromatography coupled-diode array detector

Background: Cynanchum paniculatum Kitagawa belongs to Asclepiadaceae and was used in traditional medicine to invigorate blood, alleviate edema, relieve pain, and relieve toxicity for a long time. Objective: A novel and reliable high performance liquid chromatography coupled with diode array detector method has been established for simultaneous determination of 10 bioactive compounds isolated from Cynanchum paniculatum Kitagawa, one of the herbal medicines. Materials and Methods: The chromatography analysis was performed on a SHISEIDO C18 column (S-5 µm, 4.6 mm I.D. × 250 mm) at 35oC with a gradient elution of acetonitrile and water at a flow rate of 1ml/min and UV detection at 210, 230, and 280 nm. Results: The method was validated for linearity, precision, and accuracy. All calibration curves showed good linear regression (r2 > 0.9996). Limits of detection (LOD) and limits of quantification (LOQ) fell in the ranges 0.01 - 0.28 µg/ml and 0.04 - 0.83 µg/ml, respectively. The relative standard deviation (RSD) of the intra- and inter- day test, precision test were within 1.92% and 2.43%, respectively. The mean recovery of all ranged from 92.82 to 107.96% with RSD values 0.12 - 2.18%. Conclusion: The results of validation appeared that this established method was very accurate and stabilized for the quantification of 10 bioactive compounds isolated from C. paniculatum.