Cardiovascular diseases are the most common human diseases, hence, the production of cardiological (in particular, anti-arrhythmic) medications from the natural sources is an ever-actual task. Rauwolfia serpentina Benth. is a tropical fruticose plant that is able to produce and concentrate indole alkaloids, especially ajmaline and its derivatives, which are the most effective medications against ventricular arrhythmia with low side effects. Aim of the study. Determination of the qualitative and quantitative content of indole alkaloids in cell biomass of Rauwolfia serpentina tissue culture, obtained by the prolonged in vitro growth. Materials and methods. Object: cell biomass of Rauwolfia serpentina tissue culture (K-27 strain), obtained by methods of long-term cell selection in vitro. Alkaloids content determination: TSQ Vantage LC-MS (ThermoFischer Scientific). Results. 20 indole alkaloids are found in cell biomass of Rauwolfia serpentina tissue culture (K-27 strain). The highest content is registered for ajmaline and its derivatives (0.690 % mass. for ajmaline). The contents of reserpine and yohimbine were found to be as low as 0.009 % and 0.020 %, respectively. Conclusions. It is established that the content of indole alkaloids is higher in K-27 strain in comparison to natural plant and is stable over more than 30 years of its growth. Total alkaloids content was found to be 2.8 % of dry cell biomass, and total ajmaline-type alkaloids content (including ajmaline) was found to be 1.6 % of dry cell biomass. In contrast, the total alkaloid contents in the natural plant material is reported to be in the range of 0.8–1.3 %.

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The indoline al ka loids ac cu mu la tion by R. serpentina cell lines upon sur face and sub merged main te nanceThe In sti tute of Mo lec u lar Bi ol ogy and Ge net ics, NAS of Ukraine 150, Ac a de mi cian Zabolotny Str., Kyiv, 03143, Ukraine kunakh@imbg.org.ua Indoline al ka loids ac cu mu la tion, in clud ing ajmaline, in the bio mass of R. serpentina cell line cul ture on the ex am ple of hor mone in de pend ent highly pro duc tive strain K 27 at its sur face and sub merged main te nance in nu tri ent me dia with dif fer ing min eral com po si tion has been stud ied. Op ti mal nu tri ent me dia for both ways of main te nance have been iden ti fied. The con di tions for two step main te nance in sur face, fol lowed by sub merged cul tures, in compositionally sim ple nu tri ent me dia have been spec i fied. Two step main te nance in creases al ka loids ac cu mu la tion at the early stages of growth 3 4 times and al lows de creas ing the time of cal lus tis sue grow ing from 60 80 days to 20 40 days. Keywords: ajmaline, indoline al ka loids, plant tis sue cul ture, R. ser pen tine, cell lines -al ka loids producents. Primary CallusObtained by B. G. Butenko on MC medium according to [2], green body fragment of 5 year old plant, 1964 Callus Tissue MC medium, total alkaloid content 0.1-0.3 %, ajmaline 0.04 % (1964-1968) [3] Mutagen treatment by nitrogenous yperite in 1968 M1 Mutant Line MC medium, total alkaloid content 0.2-0.48 %, ajmaline app. 0.2 % [4]Selection on nutrition medium 5C, without phytohormones according to [5] (1972-1975) Cell Line A 5C medium without phytohormones, indoline alkaloid content 0.7-0.8 %, ajmaline app. 0.4 % Mutagen treatment by ethylenimine in 1981 and the adaptation to growth on 10C medium, without phytohormones according to [8] in the course of [1982][1983][1984][1985] K 27 Strain 10C medium, indoline alkaloid content 1.2-1.8 %, ajmaline 0.9-1.2 %

A laboratory prototype of enzyme biosensor based on pH‐sensitive field‐effect transistors has been created to determine the total content of indole alkaloids in Rauwolfia serpentina tissue culture. Enzyme acetylcholinesterase was immobilized on the surface of pH‐sensitive field‐effect transistors using glutaraldehyde for covalent crosslinking with bovine serum albumin. The biosensor was characterized by high sensitivity to the total content of indole alkaloids (minimum limit of determination 0.5 μg/ml in the juice obtained from Rauwolfia serpentina tissue culture). The linear range of biosensor determination was from 2 to 15 μg/ml of the total content of indole alkaloids. The analysis of indole alkaloids using a biosensor is easy and fast; unlike traditional methods, no expensive equipment and special sample preparation are required for analysis. The created biosensor could be further used to control the total content of indole alkaloids in modern biotechnological and pharmaceutical processes of the production of drugs and dietary supplements.