OBJECTIVETo develop an HPLC-UV-ELSD method for the determination of aucubi, harpagide, harpagoside, angoroside C and cinnamic acid in Scrophularia ningpoensis.

METHODThe analytical column was SHIMADZU C18 (4.6 mm x 250 mm, 5 microm). The mobile phase was acetonirile-0.4% acetic acid in a gradient elution. Initial conditions was 5% A; 0-20 min, changed to 10% A; 20-50 min, to 55% A. The flow rate was 0.8 mL x min(-1) and the column temperature was 30 degrees C. The UV detector wavelength was set at 280 nm for harpagoside, angoroside C and cinnamic acid, and the evaporative light-scattering detector (ELSD) drift tube temperature was 105 degrees C, the flow rate of nebulizer gas was 1.2 L x min(-1) for aucubi and harpagide.

RESULTAucubi, harpagide, harpagoside, angoroside C and cinnamic acid was separated well. The linear calibration curves were obtained over of 0.752-13.536 microg for aucubi (r=0.9993, n=6), 0.8280-14.90 microg for harpagide (r=0.9994, n=6), 0.6360-11.45 microg for harpagoside (r=0.9997, n=6), 0.5440- 9.792 microg for angoroside C, (r=0.9997, n=6) and 0.0108-0.1939 microg for cinnamic acid (r=0.9999, n=6). The mean recovers of five compounds were 98.12%, 99.14%, 100.21%, 98.17% and 100.35% with RSD of 2.3%, 1.5%, 1.9%, 1.7% and 0.5%.

CONCLUSIONThis method could simultaneously determinate the content of the five compounds in the S. ningpoensis.