ABSTRACT
Echium amoenum Fisch. and C.A. Mey. [Boraginaceae] is a very popular medicinal plant which is used as a tonic, tranquillizer, diaphoretic, cough remedy, sore throat and pneumonia in Iran's traditional medicine. Callus culture of medicinal plants is one of the ways for production of secondary metabolites. In this study, callus culture of E. amoenum and its major secondary metabolite were investigated. The callus culture of E. amoenum was initiated and established from seeds in MS media with three different ratios of plant growth regulatories: kinetin, 2, 4-D and NAA. Methanolic extracts of freeze-dried calluses were compared by TLC and HPLC. The major secondary metabolite was separated by preparative HPLC and the structure of this pure compound was elucidated by UV, IR, one and two dimensional 1H and [13]C-NMR and Mass spectroscopy. Rosmarinic acid was identified by various spectroscopic methods from callus culture of E. amoenum. Rosmarinic acid is widespread within the plant cell tissue culture of the Lamiaceae and Boraginaceae families, although in insignificant quantities. Rosmarinic acid has an antimicrobial, antiviral, and anti-inflammatory effect, which makes it a valuable product for the pharmaceutical and cosmetic industries