Simultaneous quantitative determination of five alkaloids in Catharanthus roseus by HPLC-ESI-MS/MS

AIM:To establish a method to simultaneously determine the main five alkaloids of Catharanthus roseus for trace samples,a high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry(HPLC-ESI-MS/MS) analysis method was developed.METHOD:The five Catharanthus alkaloids,vinblastine,vincristine,vinleurosine,vindoline,and catharanthine were chromatographically separated on a C18 HPLC column.The mobile phase was methanol-15 nmolL-1 ammonium acetate containing 0.02% formic acid(65:35,V/V).The quantification of these alkaloids was based on the Multiple Reaction Monitoring(MRM) mode.RESULTS:This method was validated,and the results achieved the aims of the study.The intra-and inter-day precision and accuracy of the five alkaloids were within 1.2%-11.5%(RSD%) and-10.9%-10.5%(RE%).The recovery rates of the five alkaloids of samples were from 79.9% to 91.5%.The five analytes were stable at room temperature for 2 h,at 4℃ for 12 h,and at-20℃ for two weeks.The developed method was applied successfully to determine the content of the five alkaloids in three plant parts of three batches of C.roseus with a minute amount collected from three regions of China.CONCLUSION:The HPLC-ESI-MS/MS method can be used for the simultaneous determination of five important alkaloids in trace C.roseus samples.