LC-MS and LC-PDA vs. phytochemical analysis of Colchicum brachyphyllum.

In this paper, reliability and validity of a recently in-house developed dereplication strategy based on LC-MS and LC-UV/Vis PDA techniques were verified. The dereplication strategy was applied to investigate the alkaloid rich fraction of Colchicum brachyphyllum Boiss. & Haussk. ex Boiss. (Colchicaceae) which has been previously analyzed phytochemically. Both studies results, LC-MS and LC-PDA and phytochemical analysis, were matched in seven compounds namely: (-)-colchicine (5), 3-demethyl-(-)-colchicine (4), (-)-cornigerine (7), beta-lumi-(-)-colchicine (6), (-)-demecolcine (3), 3-demethyl-(-)-demecolcine (2), and 2,3-didemethyl-(-)-demecolcine (1). LC-MS and LC-PDA based dereplication strategy was not able to detect two of the compounds that were reported in the phytochemical study namely: (+)-demecolcinone and (-)-androbiphenyline. This finding could raise a question about the "naturality" of (+)-demecolcinone. On the other hand apigenin (8) was identified only using this dereplication strategy. Four compounds which had molecular ions at m/z 372 and 400 in the stems, and at m/z at 372 and 358 in flowers, could not be dereplicated, and were thus considered unknown. Their spectral data were not matched with neither the compounds that have been isolated previously from this species nor with any other colchicinoid; hence they should be pursued as potential new compounds.

Determination of (-)-demecolcine and (-)-colchicine content in selected Jordanian Colchicum species.

In our continuing attempt to investigate Jordanian Colchicum species as a potential and viable source of colchicinoids; the distributions of the most interesting colchicinoid alkaloids (-)-demecolcine and (-)-colchicine in different plant parts of wildly growing C. crocifolium Boiss., C. ritchii R. Br., and C. triphyllum Kunze, were analyzed. The method is based on the use of external reference standards and a reversed-phase gradient HPLC. While (-)-colchicine was found in all plant parts of the three investigated Colchicum species, (-)-demecolcine was not detected in the leaves and daughter corms of C. triphyllum, the corms of C. crocifolium, and the flowers, stems, and mother and daughter corms of C. ritchii. C. triphyllum was found to be the highest in total (-)-colchicine content of 0.10% (wt/wt), while C. crocifolium was the highest in total (-)-demecolcine content of 0.09% (wt/wt). During flowering, leaves and corms in C. crocifolium, and leaves and stems in C. ritchii are the main storages of (-)-colchicine. (-)-Colchicine showed a homogenous distribution in all plant parts (stems, leaves, and mother and daughter corms) of C. triphyllum of 0.10% (wt/wt). Regarding (-)-demecolcine, stems and leaves of C. crocifolium, stems of C. triphyllum, and leaves of C. ritchii are the main storages. This work reports for the first time the presence of (-)-colchicine and (-)-demecolcine in C. crocifolium.