Introduction. Wild-crafting leads to the local extinction of many medicinal plants that are rich in phenolic substances. In vitro cultivation of cells and organs of higher plants can be the optimal solution to this problem. The research objective was to study the biosynthetic activity of in vitro extracts of wild Siberian plants. Study objects and methods. The study featured callus, cell suspension, and hairy root extracts of such Siberian medicinal plants as Eleutherococcus senticosus, Codonopsis pilosula, Platanthera bifolia, and Saposhnikovia divaricata. They were obtained by in vitro cultivation using modified nutrient media of Murashige and Skoog and Gamborg. The content of secondary metabolites was studied using the methods of thin-layer and high-performance liquid chromatography. A set of in vitro experiments tested the antioxidant and antimicrobial activity of the extracts. Results and discussion. All the samples demonstrated a high content of secondary metabolites of phenolic nature. Flavonoglycosides, apigenin, and rutin were found to be the predominant biologically active substances in the callus extracts. Flavonoglycosides dominated in the suspension extracts. The root extracts contained more caffeic acid, rutin, ecdysteroids, quercetin, apigenin, cardiofolin, and coleofolide than the callus and suspension cultures. The list of prevailing secondary metabolites in the root extracts included rutin, apigenin, coleofolide, and quercetin. All the extracts showed antimicrobial and antioxidant activity. Conclusion. All the extracts demonstrated good antioxidant and antimicrobial properties. Therefore, they can be used for the production of pharmaceuticals and biologically active food supplements as they can be helpful against infectious diseases, as well as oncological, cardiovascular, and neurodegenerative diseases linked to oxidative stress.

Medicinal plants are sources of natural antioxidants. Acting as reducing agents, these substances protect the human body against oxidative stress and slow down the aging process. We aimed to study the effects of bioactive substances isolated from medicinal plants on the lifespan of Caenorhabditis elegans L. used as a model organism. High-performance liquid chromatography was applied to isolate bioactive substances from the extracts of callus, suspension, and root cultures of meadowsweet (Filipendula ulmaria L.), ginkgo (Ginkgo biloba L.), Baikal skullcap (Scutellaria baicalensis L.), red clover (Trifolium pretense L.), alfalfa (Medicágo sativa L.), and thyme (Thymus vulgaris L.). Their effect on the lifespan of C. elegans nematodes was determined by counting live nematodes treated with their concentrations of 10, 50, 100, and 200 µmol/L after 61 days of the experiment. The results were recorded using IR spectrometry. The isolated bioactive substances were at least 95% pure. We found that the studied concentrations of trans-cinnamic acid, baicalin, rutin, ursolic acid, and magniferin did not significantly increase the lifespan of the nematodes. Naringenin increased their lifespan by an average of 27.3% during days 8–26. Chlorogenic acid at a concentration of 100 µmol/L increased the lifespan of C. elegans by 27.7%. Ginkgo-based kaempferol and quercetin, as well as red clover-based biochanin A at the concentrations of 200, 10, and 100 µmol/L, respectively, increased the lifespan of the nematodes by 30.6, 41.9, and 45.2%, respectively. The bioactive substances produced from callus, root, and suspension cultures of the above medicinal plants had a positive effect on the lifespan of C. elegans nematodes. This confirms their geroprotective properties and allows them to be used as anti-aging agents.

Comfrey (Symphytum officinale) root preparations are known for their analgesic and anti-inflammatory properties. In this study, the phenolic profile of extract samples of Symphytum officinale callus root cultures (up to 70% of ethanol) was determined using various approaches. For the first time, a unique polymer of two types was isolated from samples of comfrey root cultures (the caffeic acid derivative is a new class of natural polyether with 3-(3,4-dihydroxyphenyl) glyceric acid as a repeating unit). M-methoxybenzoic acid (30.05 μg/mL extract) or rosmarinic acid (45.70 μL/mL extract) were identified as the main phenolic component. Rosmarinic, chlorogenic, and caffeic acids have anti-inflammatory, anti-apoptotic, antitumor, neuroprotective, antioxidant, and other properties, which also determine the characteristics of comfrey and preparations made from it. The studied phenolic profile and the analysis of published data showed that the extract samples of comfrey callus cultures had similar characteristics, which makes them an important source of phenolic compounds with pronounced antioxidant activity.

The work reveals the results of studying the content of biologically active substances in samples of extracts of Ginkgo biloba callus cultures. Callus cultures grown in vitro on liquid nutrient media were the objects of the study. Considering various factors affecting the yield of the target components during extraction, the volume fraction of the organic modifier in the extracting mixture, the temperature factor, and the exposure time were identified as the main ones. The maximum yield of extractive substances (target biologically active substances with a degree of extraction of at least 50%) from the samples of callus culture extracts was detected at a ratio of extragent of 70% ethanol, a temperature of 50 °C, and exposure time of 6 h. Flavonoids, such as luteolin, quercetin, isoramentin, kaempferol, and amentoflavone, were isolated in the extract samples. As a result of column chromatography, fractions of individual biologically active substances (bilobalide, ginkgolide A, B, and C) were determined. The proposed schemes are focused on preserving the nativity while ensuring maximum purification from associated (ballast) components. Sorbents (Sephadex LH-20, poly-amide, silica gel) were used in successive stages of chromatography with rechromatography. The degree of purity of individually isolated substances was at least 95%.