Screening of Catharanthus roseus secondary metabolites by high-performance liquid chromatography

Two direct HPLC analytical methods for the screening of the major indole alkaloids of Catharanthus roseus hairy roots and their iridoid precursors have been developed. Photodiode array and fluorescence detection were performed. The separation was achieved on a reversed-phase C18 column. The first method allowed the separation of catharanthine, serpentine, tabersonine, vindoline, vinblastine, and vincristine in 20 min. Ajmalicine, tryptophan, tryptamine and secologanine were separated using the second method in 13 min. The identification of the compounds was based on the retention time and the comparison of UV spectra with those of authentic standards. A simplified alkaloid extraction method was developed in order to accelerate sample preparation. The assays were successfully used to quantify major compounds of the secondary metabolism of hairy root cultures of C. roseus, thus providing a reliable tool for rapid screening of C. roseus secondary metabolite samples. In these cultures, ajmalicine, serpentine, catharanthine, tabersonine, and tryptamine were detected, but tryptophan, vindoline, vinblastine and vincristine were not.