In vitro cultures, at different stages of morphogenesis, were established from leaves of Leucojum aestivum (Amaryllidaceae) for determination of their galanthamine content, an alkaloid that possesses cholinesterase inhibitory activity and which is used for the treatment of Alzheimer's disease. A suitable HPLC method for qualitative and quantitative determination of galanthamine in both in vitro and in vivo extracts has been developed. Confirmation of the identity of galanthamine in sample extracts was achieved using LC-MS-MS. A correlation was observed between the state of differentiation and the galanthamine content of the tissue cultures. No galanthamine was detected in the roots grown in vitro, while all bulblets grown in vitro showed the presence of this alkaloid, with dramatic variations in concentration levels, according to the growth substance balance. The best result (6.79 × 10−3 % of D.W.) was obtained with bulblets initiated with NAA (10 μM) combined with BA (0.5 μM).
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