ИСТИНА

For thousands of years, herbal medicines made from terrestrial plants were used by humans to prevent and treat diseases. Plants contain a broad range of bioactive compounds such as alkaloids, phenolics, triterpenoid and steroidal glycosides etc. Plant cell culture is a perspective material for isolation of biologically active compounds. Thus, cell biomass grown in bioreactors can be used to produce steroidal glycosides. D. deltoidea plant cell culture contains an extremely complicated mixture of furostanol steroidal glycosides. Therefore, it is critical to control the composition of cell cultures for medical use. Sample preparation is the first and crucial step in the analysis of medical plants and cell culture, because it is necessary to extract the desired chemical components from plant material for further separation and characterization. Optimization of extraction conditions requires varying many factors that may influence the experiment. Experimental design is usually used for reducing the number of the experiments, minimizing the effects of uncontrolled factors and revealing the most and less influencing factors. The ultrasound assisted extraction (UAE) method for isolation of steroidal glycosides from D. deltoidea plant cell suspension culture with a subsequent HPLC-MS determination was developed. After the organic solvent was selected via a two-factor experiment the optimization of extraction parameters was carried out using Latin Square 4×4 experimental design for the following parameters: extraction time, organic solvent concentration in extraction solution and the ratio of solvent to sample. It was also shown that UAE method is not suitable for isolation of steroidal glycosides from the D. deltoidea plant material. The results were confirmed using the multiple successive extraction method and refluxing extraction. Optimal conditions for the extraction of steroidal glycosides by UAE method were: extraction time, 60 min; acetonitrile (water) concentration, 50%; the ratio of solvent to sample, 400 mL/g. Also, the developed method was tested on D. deltoidea cell suspension cultures of different terms and conditions of cultivation.