Determination of Vincristine and Vinblastine in Catharanthus roseus Plants by High Performance Liquid Chromatography/ Electrospray Ionization Mass Spectrometry

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Abstract
A reversed phase high performance liquid chromatographic/ mass spectrometric (LC-MS) method with an electrospray ion source was used in this study to determine the concentration of two dimeric alkaloids, vinblastine and vincristine, in nearisogenic lines of Catharanthus leaves. A linear gradient of 25 mM ammonium acetate in methanol (solution A) and the same concentration in water (solution B) served as the mobile phase. A C18 column was used to separate those two compounds from other components in the extracted plant samples. Mass spectra generated by the electrospray ionization source were dominated by protonated molecules with little or no fragmentation being observed under the experimental conditions applied. Since very little fragmentation occurred. the method exhibited enhanced sensitivity over other LC-MS techniques. Product-ion mass spectra of vincristine and vinblastine were produced in a single quadrupole mass spectrometer through collision-induced dissociation (CID) between the capillary exit and first skimmer of the electrospray source. CID spectra can be used to unequivocally differentiate between vincristine, vinblastine and other alkaloids.

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