Simultaneous determination of scopolamine, hyoscyamine and anisodamine in in vitro growth media of selected Solanaceae hairy roots by CE method

• Simple method development process resulted in extremely rapid separation of selected tropane alkaloids with very high efficiency was developed.
• Quantification of scopolamine, hyoscyamine and anisodamine in plant cultures growth media without any pretretment step was presented.
• Impact of frequency of detector probing on the peaks parameters in CE has been demonstrated and discussed for the first time.

An electrophoretic method for fast separation of three tropane alkaloids (hyoscyamine, anisodamine and scopolamine) was presented. The substances were complete resolved in less than one minute due to utilization of relatively short capillary (20.2 cm effective length) and high voltage (25 kV). Detector probing frequency was found among the parameters that significantly affected the detection sensitivity. The performed experiments showed insufficient available probing frequency of used commercial spectrophotometric detector according to capillary electrophoresis (CE) separation potential.Under the optimized conditions the background electrolyte (BGE) was composed of 20 mM Tris, 6 mM HCl and 20 mM NaCl (pH 8.50). All analyses were carried out in fused silica capillaries of 50 μm (inner diameter) and 31.2 cm (total capillary length). Samples were injected hydrodynamically (5 s; 3.45 kPa) without any sample preparation step and separation was performed at 25 kV. The elaborated method was applied in plant cultures growth media analysis after incubation with hairy roots of selected Solanaceae species. The performed experiments proved the usefulness of CE in quality control of biotechnological processes.