Isolation of Individual Compounds from the Aerial Part of &lt;i&gt;Comarum palustre &lt;/i&gt;L. and Their Structure Elucidation Using Spectroscopic Methods

Introduction. Plant biologically active substances (BAS) are valuable for drug development in view of their pharmacological effects and metabolic proximity to the human body. As an example of a useful herbal raw material, the roots of Scutellaria baicalensis, which contain species-specific flavonoids, baicalin and scutellarin, possess the potency of creating antiviral drugs and functional nutrition systems with antioxidant and adaptogenic properties. In order to levelling the existing limitations of using intact plant in pharmaceutical practice, we proposed a nature-saving and ergonomic approach of cultivation of S. baicalensis cells under in vitro conditions. This technology allows to extract active substances of S. baicalensis without causing damage to the environment and contributes to the reduction of the time required to produce the necessary amount of plant material with an increase in the efficiency of production processes.Aim. The development of dry extract technology from S. baicalensis callus biomass.Materials and methods. The object of the study – biomass of S. baicalensis cells obtained from callus culture. Functional characteristics (friability, Hausner’s coefficient, Carr’s index, fractional composition, as well as porosity and hygroscopicity) of S. baicalensis biomass and dry extract on its basis were determined according to the methods of the EAEU Pharmacopoeia. Extraction of plant material was carried out by maceration method with heating in a water bath with reflux condenser. Qualitative and quantitative analysis of BAS in the dry extract was carried out by spectrophotometry and high-performance liquid chromatography. Quality indicators and functional characteristics of the finished product were evaluated in accordance with pharmacopeial requirements.Results and discussion. The functional characteristics of the S. baicalensis biomass were evaluated and the possibility of obtaining a dry extract on its basis was confirmed. We selected parameters of BAS extraction process, established the presence of baicalin in the biomass extract and carried out a comparative evaluation of aglycone profile in the extract based on callus culture and roots of intact plant. The properties of the finished product were evaluated for compliance with regulatory requirements, as well as technological characteristics.Conclusion. The quality indicators of the dry extract of the S. baicalensis biomass satisfy the normative conditions. The technological characteristics of dry extract are suitable for the utilisation as a phytosubstance for the development of medicines of combined type and dietary supplements.

Section: методы и оборудованиеunclassified

Section: Introductionunclassified

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The Development of Dry Extract Technology from Callus Culture Biomass of Scutellaria baicalensis

Danilova,

Pivovarova,

Flisyuk

2023

“…NMR spectroscopy is approved for quality control of active pharmaceutical ingredients (APIs) by numerous regulatory guides, including the pharmacopeias of Russia, the Eurasian Economic Union, the European Union, and the United States of America. It is used in pharmaceutical practice for identification [11], qualitative control [12], and determination of impurities [13] and residual organic solvents. There are two ways of compound identification by NMR spectroscopy: by spectral data interpretation and via comparison of the obtained NMR spectrum with a reference.…”

Section: Introductionmentioning

confidence: 99%

Analysis of cis-isomer-enriched dihydroquercetin sample by 1D and 2D NMR spectroscopy

Terekhov,

Taldaev,

Bocharov

et al. 2024

Introduction. The structure of dihydroquercetin (DHQ) is characterized by two chiral centers at positions 2 and 3 of the benzopyran cycle, resulting in possible diastereomers: trans- and cis-isomers. Therefore, the development of methods for qualitative and quantitative control of DHQ diastereomers in analyzed samples is essential for patient safety management. Nuclear magnetic resonance (NMR) spectroscopy is one of the physicochemical methods that can be used for this purpose.Aim. The study objective was to accumulate the analytical and structural characteristics of cis-DHQ by NMR spectroscopy of the spheroidal form of this flavonoid (DHQs).Materials and Methods. 1D 1H, 1H,1H-COSY, 1H,1H-NOESY, and 1H,13C-HSQC NMR spectra were acquired at 298 K on an 800 MHz NMR spectrometer equipped with a TXI triple resonance probe. The number of scans was 32. The mixing time in the NOESY experiment was 400 ms. The 1H and 13C were analyzed using CcpNmr software. The dihedral angles were calculated by applying the Karplus equation.Results and discussion. In trans-DHQ, the chemical shift values for H2 and H3 are 4.93 ppm and 4.52 ppm, respectively, and in cis-DHQ they are 5.31 ppm and 4.20 ppm, respectively. The spin-spin coupling constants between H2 and H3 of trans- and cis-DHQ are 12.00 Hz and 2.40 Hz, respectively. Thus, the dihedral angles for the trans- and cis-isomers are 154° and 64°, respectively. We found that DHQs contains 12.5 % of the cis-isomer.Conclusion. Our experiments confirmed that NMR spectroscopy can discriminate between trans- and cis-DHQ based on the chemical shift values for the cross-peaks of H2 and H3. The second major finding was that this method can be considered as a more selective quantitative analysis than HPLC with UV detection without reference. One of the most important results of this study for drug development is the updated information on the structural parameters of DHQ diastereomers in the liquid phase.

Antimicrobial and Antioxidant Activity Screening of Mushrooms Growing in the Leningrad Region

Whaley,

Novikova

et al. 2023

Introduction. The emergence of new strains of microorganisms that are multidrug resistant (MDR) in relation to the antimicrobial drugs used is one of the pressing problems of modern medicine. To prevent an increase in MDR-related deaths, the search for new antibiotics and their introduction into medical practice must be continuously ongoing. Infectious diseases are also accompanied by cell damage and the development of free radical oxidation processes, therefore the search for new antioxidants is also an important task. Considering the powerful biosynthetic potential of basidiomycetes, this group of fungi has every prospect of becoming a new source of biologically active substances in general, as well as antibiotics and antioxidants in particular. Cap mushrooms, represented mainly by basidiomycetes, number about 14,000 species and are an accessible source of raw materials for the search for promising antimicrobial compounds and antioxidants.Aim. Study of the antioxidant and antimicrobial activity of total extracts obtained from cap mushrooms against Escherichia coli, Staphylococcus aureus and Candida albicans and assessment of the suitability of cap mushrooms as a natural source of substances with antimicrobial and antioxidant activity.Materials and methods. The antifungal and antibacterial activity of the extracts was determined by the micromethod of two-fold serial dilutions in a liquid nutrient medium in 96-well plates in duplicate. The study of this type of biological activity was carried out against reference (type) strains Staphylococcus aureus ATCC 6538-P, Escherichia coli ATCC 25922, Candida albicans NCTC 885-653. To study antioxidant activity using DPPH, we used alcoholic extracts from the fruiting bodies of mushrooms obtained by maceration with 96 % ethanol at a ratio of raw materials to extractant of 1 to 8 for 24 hours, an aqueous solution of ascorbic acid (vitamin C) and an ethanol solution of Trolox.Result and discussion. In relation to S. aureus, a representative of gram-positive flora, the studied extracts of cap mushrooms showed low activity, on average about 2500 or 5000 μg/ml. In relation to E. coli, a representative of gram-negative flora, 8 % of the studied cap mushroom extracts showed an average activity of about 1250 μg/ml. The largest number of cap mushroom extracts – 19% of all studied species – showed activity against the yeast micromycete C. albicans. The highest activity against C. albicans was observed in extracts of the mushrooms Cantharellula umbonata with an MIC of 625 μg/ml, Cortinarius olivaceofuscus with an MIC of 625 μg/ml, and Hypomyces chrysospermus with an MIC of 312 μg/ml. During screening of antioxidant activity, the studied extracts were divided into three groups: with high (more than 50 % PPR), medium (from 15 to 50 % PPR) and low (less than 15 %) antioxidant activity. It was shown that the sum of phenolic compounds significantly correlates with the level of antioxidant activity in all three groups, but in the groups with medium and low antioxidant activity there are also other non-phenolic groups of compounds that make a significant contribution to the total antioxidant activity.Conclusion. Cap mushrooms are a promising source of biologically active substances with antifungal and antioxidant activity.