Sesquiterpene pyridine alkaloids are considered to be the active components of Tripterygium wilfordii Hook. f. A rapid method was developed for comprehensive profiling of sesquiterpene pyridine alkaloids from an extract of root bark of T. wilfordii using supercritical fluid chromatography-diode array detector-tandem mass spectrometry (SFC-DAD-MS/MS). Alkaloids were separated on a BEH 2EP column within 10min, eluted by CO2-methanol as mobile phase with a back pressure of 13.8 Mpa and a column temperature of 45 degrees C. MS/MS analysis of [M + H](+). ion of each alkaloid standard showed that all the pyridine alkaloids produced very similar fragmentation patterns. The product-ions at m/z 206 and 204 were identified as the diagnostic fragments while mass region of 200-500 Da was assigned as the characteristic region. As a result, 71 components in the extract were identified as sesquiterpene pyridine alkaloids, including 40 wilfordate/evoninate type alkaloids, 13 iso-wilfordate/evoninate type alkaloids and 19 hydroxyl-wilfordate/evoninate type alkaloids. The results proved the feasibility of SFC-DAD-MS/MS method for the rapid and high-throughput analysis of sesquiterpene pyridine alkaloids in complex samples. (C) 2015 Elsevier B.V. All rights reserved.