Direct determination of fourteen underivatized amino acids from Whitmania pigra by using liquid chromatography-evaporative light scattering detection

A new reversed phase high performance liquid chromatography-evaporative light scattering detection (RP-HPLC-ELSD) method has been developed for the direct determination of fourteen amino acids in Whitmania pigra, a commonly used traditional Chinese medicine (TCM). Simultaneous separation of these fourteen amino acids was achieved on a Prevail C18 column. Under the condition of gradient elution, the fourteen amino acids were separated within 25 min. The drift tube temperature of ELSD was set at 115 °C, and with the nitrogen flow rate of 2.5 l/min. All calibration curves showed good linear regression (r > 0.9975) within test ranges. The recoveries ranged from 94.8% to 104.4%. The limit of detection (LOD) for each compound was more than 20 mg/l at a signal to noise ratio (S/N) of 3. The validated method was successfully applied to quantify fourteen amino acids from Whitmania pigra, which provides a new basis of overall assessment on quality of Whitmania pigra.