Simultaneous Determination of Osthol and Columbianadin in Tobacco Flavor by RP-UPLC

To establish a method for simultaneous determination of osthol and columbianadin in tobacco flavor by reversed phase ultra performance liquid chromatography,20 kinds of tobacco flavor samples were extracted by acetionitrile-water(volume ratio of 1∶1) and separated at 30 ℃ on ACQUITY UPLCBEH C18(2.1 mm i.d.×50 mm,1.7 μm) column eluted with acetonitrile-water as the mobile phase.Flow rate was set at 0.3 mL/ min and the detection wavelength was set at 324 nm.The results indicated that: The chromatographic peak of osthol and columbianadin standard samples were sharp and symmetric.The resolution of these two substances were greater than 1.5.The calibration curve of osthol and columbianadin were linear within the range from 0.20 to 10.00 mg/L(r=0.9999);Osthol and columbianadin were not detected in 20 kinds of tobacco flavor;When osthol and columbianadin were added into blank samples at high,middle and low levels,the average recoveries of osthol were from 98.00 % to 100.44 %,with RSD of 0.74 % to 2.54 % in three days.The average recoveries of columbianadin were from 99.41 % to 101.02 %,with RSD of 0.92 % to 2.27 % in three days.