Simultaneous determination of scopolamine, hyoscyamine and littorine in plants and different hairy root clones of Hyoscyamus muticus by micellar electrokinetic chromatography

Hyoscyamus muticus hairy root clones were established following infection w ith Agrobacterium rhizogenes strains A4, LBA-9402 and 15834 and with A. tum efaciens strain C58C1pRTGus104. The accumulation of tropane alkaloids hyosc yamine, littorine and scopolamine was evaluated by micellar electrokinetic capillary electrophoresis. Littorine was reported for the first time in the se clones as well as in the roots of the intact plant and confirmed by coll ision induced dissociation-mass spectrometry. Tropane alkaloid content in h airy roots was compared with leaves and roots of normal plants at two veget ative stages. Significant differences appeared between the alkaloid content s of the different crones. In particular, all the hairy root clones and the roots of the intact plant produced 1.5-3 and 4.5-9 times more littorine th an scopolamine, respectively. The only exception was clone KB7, carrying th e h6h gene, which overproduced scopolamine. The aerial parts of H. muticus plants did not contain any littorine, thus indicating different transportat ion or translocation mechanisms of the various tropane alkaloids. (C) 2000 Elsevier Science Ltd. All rights reserved.