ИСТИНА

Introduction Many of steroidal saponins have anticancer potential and may be used for treatment of cardiovascular diseases. Cell biomass grown in bioreactors can be used to produce steroidal saponins. Dioscorea deltoidea plant cell culture contain an extremely complicated mixture of furostanol steroidal saponins. Therefore, it is critical to control the composition of cell cultures for medical use. Methods For the extraction of steroidal saponins ultrasound assisted extraction (UAE), multiple successive extraction (MSEM) and reflux extraction (RE) methods were used. LC separation was performed on a reversed phase C18 column in gradient elution mode. MS detection was conducted in the positive electrospray ionization mode (SIM mode). Results Optimization of the UAE parameters was performed with use of Latin square experimental design [1]. The following parameters have been optimized: extraction time, organic solvent concentration in extraction solution and the ratio of solvent to sample. Optimal conditions for the extraction of steroidal saponins by the UAE method were: extraction time, 60 min; acetonitrile (water) concentration in extraction solution, 50% (50%); the ratio of solvent to sample, 400 mL/g. As a result, high degree of the extraction of steroidal saponins from the D. deltoidea plant cell suspension culture was achieved. The obtained results were confirmed by MSEM and RE. It was also shown that the UAE method is not suitable for isolation of steroidal glycosides from the D. deltoidea plant material. Conclusions For the determination of steroidal saponins from D. deltoidea cell culture comprehensive and representative pre-LC-MS extraction procedure was proposed. Low degree of the steroidal saponins extraction from the plant material is most likely due to the lack of mechanical tissues in the cell culture, in contrast to plant material. The UAE method was tested on a series of 18 cell culture samples of different inoculation date, growing condition and growth period. Acknowledgements This work was supported by Russian Science Foundation (Grant No. 17-13-01146) for Moscow State University. Novel Aspect Comparison of the different approaches performance characteristics have been carried out with respect to finding an optimal pre-LC-MS treatment of D. deltoidea cell culture. References [1] Sarvin B., Fedorova E., Shpigun O., et al., Journal of Chromatography B, 1080, 64-70 (2018).