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The greater celandine (Chelidonium majus L.) has been known for the centuries as a medicinal plant. One of the therapeutic agents based on C. majus is anticancer drug Ukrain™ known as a semi-synthetic C. majus alkaloid derivative. Although there are no doubts about antitumor properties of the drug, there is still controversy about its composition. In this study, Ukrain™ was subjected to TLC and LC-MS/MS analyses to compare it with C. majus alkaloid root extract and to determine its composition. Moreover, microbiological activity of both Ukrain™ and the alkaloid extract were tested against Bacillus subtilis strains using TLC-direct bioautography. Sanguinarine, chelidonine, α-homochelidonie and chelerythrine were found to have antibacterial properties. Besides chelidonine, sanguinarine, chelerythrine, protopine, allocryptopine, homochelidonie, berberine and coptisine reported earlier in literature, the presence of stylopine, norchelidonine, dihydrochelidonine and hydroberberine in Ukrain™ was detected, and here they have been reported for the first time.

The presented paper is the first to show thin layer chromatography (TLC) analysis based on eutectic mobile phases (Deep Eutectic Solvents – DES). During the experiment 25 eutectic mixtures were investigated for their chromatographic properties. Most of them belong to the natural deep eutectic solvents (NADES) group. Also, new eutectic liquids based on phenolics and terpenes, not previously employed in analytical practice, were tested. The eutectic liquids were investigated as pure or diluted with solvents used in chromatographic routine: methanol, water, acetone, chloroform or diethyl ether. The analyses were carried out using classic and high performance silica gel plates. The working solution was a mixture of five alkaloids found in genus Chelidonium, namely sanguinarine, coptisine, chelerythrine, chelidonine, and berberine, with UV light detection of 366 nm. This report proves that eutectic TLC is possible and that the eutectic interactions play a crucial role in the separation process. In most of the tested modifications at least partial separation was achieved. The most successful mobile phase, which enabled separation of all the tested alkaloids, was the equimolar mixture of menthol and phenol with a 35% addition of methanol. The system was also effective in separating alkaloids in the real Chelidonium maius extract sample.

In this study, we introduced a simple and sensitive method of capillary electrophoresis with ultraviolet light-emitting diode-induced native fluorescence (UV-LEDIF) detection for the determination of isoquinoline alkaloids in extracts of Chelidonium majus L. Samples were extracted with acidic methanol and the extracts were directly analysed by CE. Simultaneous determination of protopine, chelidonine, coptisine, sanguinarine, allocryptopine, chelerythrine and stylopine was performed in 20 mM phosphate buffer (pH 3.1). The baseline separation of these alkaloids was finished within 20 min. As these alkaloids have native fluorescence, they were directly detected using the commercially available UV light emitting diode without troublesome fluorescent derivatisation. Satisfactory LOD values were obtained for the studied compounds considering their appearance in natural extracts. Lower limits of detection were 0.05 μg/mL for protopine, 0.06 μg/mL for stylopine and allocryptopine, 0.07 μg/mL for chelidonine, 0.22 μg/mL for sanguinarine, 1.7 μg/mL for chelerythrine and 5.5 μg/mL for coptisine. The developed method was successfully applied to determine the contents of seven alkaloids in the aerial parts of Chelidonium majus L, which varied from 0.025 to 0.763% (w/w). Also, to demonstrate the potential of the proposed CE method, an estimation of the cytotoxic properties of selected Celandine alkaloids in a natural extract was carried out.

A simple and effective method for the sensitive detection of alkaloids in crude plant extracts applying capillary electrophoresis with field amplified sample injection (FASI) is described. This method was compared with normal pressure injection for the determination of alkaloids in methanolic extracts from roots of Berberis vulgaris L. (Berberidaceae) and Hydrastis canadensis L. (Ranunculaceae) using a 1:1 mixture of 200 mM ammonium acetate at pH 3.1 and methanol. By introducing a short plug of 70% methanol (v/v) before electrokinetic injection with 16 kV for 8 s the concentration sensitivity was 1000-times higher compared to hydrodynamic injection for 1 s. No difference between both injection methods for selectivity and resolution of the obtained electropherograms was found. The influence of voltage and injection time on the introduced sample amount was investigated using a mixture of berberine and chelidonine as model substances.

A simple and precise method using ion-pair high-performance liquid chromatography was developed for the simultaneous determination of eight isoquinoline alkaloids, namely chelidonine, berberine, protopine, coptisine, tetrahydrocoptisine, 6-methoxydihydrochelerythrine, 6-methoxydihydrosanguinarine and dihydrosanguinarine, in Chelidonium majus L.. A reversed-phase system consisting of a chemically bonded ODS silica gel column and 0.05 M tartaric acid—methanol—acetonitrile (44:10:46) containing 0.5% sodium dodecyl sulphate as the mobile phase was used. The eight alkaloids were completely separated within 40 min. The analytical results for various samples are presented.