Vol.:(0123456789) 1 3 Chromatographia https://doi.org/10.1007/s10337-020-03977-x ORIGINAL Aporphine Alkaloids from Triclisia dictyophylla Diels by pH‑Zone Refining Countercurrent Chromatography Eutrophe Le Doux Kamto 1,2  · Simony C. Mendonça 2  · Auguste Abouem A Zintchem 3  · Eric Robert Tiam 1  · Dieudonné Emmanuel Pegnyemb 1  · Gilda Guimarães Leitão 2 Received: 10 July 2020 / Revised: 2 October 2020 / Accepted: 10 October 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020 Abstract A new dimeric aporphine alkaloid, biscocsarmine, 2, together with lirioferine, 1, and cocsarmine, 3, were isolated from the ethanol extract of the stems of Triclisia dictyophylla Diels (Menispermaceae) using a preparative pH-zone refining CCC in a single step. The first solvent system family tested for this fractionation was hexane–ethyl acetate–methanol–water (HEMWat) at a series of solvent ratios, and hydrochloric acid and triethylamine at 5 mM as eluter/retainer. None of these HEMWat ratios, however, gave suitable K for the target alkaloids. Ethyl acetate–n-butanol–water (EBuWat) was tested instead and the selected solvent system was composed of ethyl acetate–n-butanol–water 5:5:10 (v/v), where triethylamine (60 mM) was added to the upper organic stationary phase as a retainer and hydrochloric acid (5 mM) to the aqueous mobile phase as an eluter. The chemical structures of the isolated compounds were elucidated based on comprehensive spectroscopic and spectrometric techniques. This is the first report of aporphine alkaloids in this plant species. Keywords Aporphine alkaloids · Menispermaceae · pH-Zone refining CCC · Quaternary alkaloids · Gradient elution Abbreviations EBuWat Ethyl acetate–n-butanol–water HEMWa Hexane–ethyl acetate–methanol–water TEA Trimethylamine Introduction Triclisia dictyophylla Diels (synonym Tiliacora flava Exell, Tiliacora giletti (De will)), Menispermaceae, is a climbing plant or scrambling shrub of the lowland dense rain forest with stems that can be up to 30 m long. The plant occurs from Liberia to Central Africa, Democratic Republic of Congo, Angola and Tanzania [1]. T. dictyophylla is widely used in African medicine for the treatment of anemia, con- vulsive coughing, diarrhea, epileptic attacks, fever, joint pain, heart palpitations, leprosy, malaria, edema, purulent catarrh, stomach problems and venereal diseases [2–4]. Early phytochemical studies on this plant genus revealed the presence of tertiary and quaternary alkaloids including bisbenzylisoquinolines, an oxo-isoquinoline, an indenoiso- quinoline and a morphinan-type alkaloid [4–7]. Previous pharmacological reports revealed that the root extract of T. dictyophylla exhibited strong anti-plasmodial, anticoagula- tion, antithrombotic and anti-tussive activities [3, 8–12], making this plant a potential source of bioactive compounds. Conventional methods for separating and purifying alkaloids remain a challenge, regarding the multiple chromatographic steps to be achieved. High-speed countercurrent chromatog- raphy (HSCCC) is a liquid–liquid partition chromatography technique without a solid support matrix where a wide range Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10337-020-03977-x) contains supplementary material, which is available to authorized users. * Eutrophe Le Doux Kamto lekamto@gmail.com * Gilda Guimarães Leitão ggleitao@ippn.ufrj.br; ggleitao@yahoo.com.br 1 Department of Organic Chemistry, Faculty of Science, University of Yaounde 1, Yaounde, Cameroon 2 Instituto de Pesquisas de Produtos Naturais, Centro de Ciencias da Saúde, Bl. H, Universidade Federal do Rio de Janeiro, Ilha do Fundão, Rio de Janeiro, RJ 21.941-902, Brazil 3 Department of Chemistry, Higher Teacher Training College, University of Yaounde 1, Yaounde, Cameroon