Abstract:

A method of reversed-phase high performance liquid chromatography (HPLC) coupled with evaporative light scattering detection (ELSD) was developed for the determination of jervine and veratramine in veratrum plants. The extraction method of total active alkaloids from the raw material was also established. The separation and quantification were achieved using a Kromasil C18column (250 mm×4.6 mm, 5 μm), and a mobile phase of acetonitrile and 0.1% trifluoroacetic acid with the following gradient elution: 20%acetonitrile at the first 5 min, 20%-40%acetonitrile at the 5-30 min, 40%-20%acetonitrile at the 30-40 min, 20%acetonitrile at the 40-45 min with a flow rate of 0.8 mL/min; column temperature of 35 ℃ and monitored by an ELSD detector with the drift tube of 98 ℃ and the nitrogen flow rate of 2.2 L/min. The calibration curves for jervine and veratramine were linear over the ranges of 42.05-980 mg/L and 43.52-1020 mg/L, respectively. The recoveries were 99.2% and 101.4% with relative standard deviations of 1.7% and 2.1% (n=6), respectively. The limits of detection for jervine and veratramine in raw material were 18.37 mg/kg and 21.50 mg/kg, respectively, with 3 times of the signal to noise ratio. This HPLC-ELSD method is rapid, simple, accurate and convenient. It can be used as one of the direct and reliable means for quantitative determination of the active alkaloids in veratrum plants.

Key words: jervine, veratramine, veratrum plants , high performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD)