The purpose of this work was the studying and modeling of the extraction properties of the sorbitol-based natural deep eutectic solvent (NADES) and sorbitol-based solvents in regard to biologically-active substances (BASs) from Glycyrrhizae roots using theoretical fundamentals based on the laws of statistical physics, thermodynamics, and physical chemistry previously developed by us. In our studies, we used Glycyrrhizae roots, simple maceration, plant raw material:solvent ratio 1:10 w/v, temperature 25 °C, extraction time 24 h; standards of licuroside and glycyram; RP HPLC, differential scanning calorimetry, integral dielectric, impedance and conductivity spectroscopy method of analysis; the following solvents: sorbitol-based NADES sorbitol:malic acid:water (1:1:3 in molar ratio), a modified solvent based on NADES sorbitol:malic acid:water:glycerin (1:1:1:1 in molar ratio) and sorbitol-based solvents sorbitol:ethanol:water at different ratios. It has been found that regression equations for sorbitol-based solvents in coordinates predicted by the theory have a high value of determination coefficient that equals to R2e = 0.993 for glycyram and R2e = 0.976 for licuroside. It has been found that the extraction properties of sorbitol-based NADES with a dielectric constant (ε) equal to 33 ± 2 units are equivalent to those of the sorbitol:ethanol:water solvent with ε = 34 units, and the extraction properties of modified solvent based on NADES with ε = 41 ± 2 units are inferior to those of the sorbitol-ethanol-water solvents with maximum value of BASs yield with the dielectric constant range 40 ÷ 50 units. The theoretical fundamentals suggested provide a possibility for an explanation of the mechanism, quantitative description of the extraction properties of the solvent, and target search of the optimal solvent by its dielectric constant.

The goal of this work was the studying and modelling of the extraction properties of fluoroorganic solvents in regard to low-polar biologically active substances (BAS) from plant raw materials (PRM). Anethum graveolens L., Foeniculum vulgare Mill., Pastinaca sativa L., Pimpinella anisum L., Silybum marianum L. fruits, Curcuma longa L. roots, Eucalyptus viminalis Labill., Laurus nobilis L. leaves, Hypericum perforatum L. herbs, and Syzygium aromaticum L. buds were used. As solvents, we used Novec 1230, Novec 7100, MR6S4, and R141b. Quantitative and qualitative analyses of BAS were carried out by RP HPLC and GC-MS. We also used a circulating method in Soxhlet apparatus. A mathematical model that quantitatively describes the extraction properties of fluoroorganic solvents using molecular descriptors such as the relative fraction of fluorine atoms in the molecule of the fluoroorganic solvent and LTPSA has been developed.

Objective: The objective of this work was to carry out studies in the field of development of a hydroalcoholic extract from the liquorice root that has a high level of antimicrobial activity.Methods: For the antibacterial study of extracts, we have used the agar well diffusion method. In our research, we have utilized six test-strain microorganisms: Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Proteus vulgaris ATCC 4636, Pseudomonas aeruginosa ATCC 27853, Candida albicans ATCC 885/653, and Bacillus subtilis ATCC 6633. Authors have applied vector theory for antimicrobial activity comparison of liquorice root extracts.Results: Phytochemical composition and antimicrobial activity of hydroalcoholic extracts from the liquorice root with the use of different concentrations of ethanol have been studied. The optimal range of ethanol concentration of 65±15% v/v has been found. The dependency between the integral index of antimicrobial activity of the extract and concentration of glycyram and licuroside has been found. The minimal concentration of glycyram and licuroside in the extract has been calculated for an exhibition of the target level of extract’s antimicrobial activity, which is 1.0 and 0.11% w/v, respectively.Conclusion: It has been found that the concentration of licuroside has greater influence on the integral index of extract’s antimicrobial activity in comparison with that one of glycyram. A highly effective technology for obtaining an extract with target phytochemical parameters has been suggested.

The purpose of this study was to develop a method of qualitative and quantitative assessment of biologically active substances of P. fruticosa L. herb. As object of study was used P. fruticosa L. herb. We used the method of high performance liquid chromatography in reversed-phase variant. As a result of chromatography, it was found that the chromatographic system used can effectively separate and identify the polyphenol complex of the herb P. fruticosus L. The chromatogram shows that the polyphenol complex of the herb P. fruticosus L. consists of the glycosides quercetin, kaempferol, rhamnetin, procyanidins, ellagotannins and ellagic acid and chlorogenic acid. Aglycones were found in the hydrolysis products: quercetin, kempferol, ellagic acid, and trace amounts of rhamnetin. Thus, as a result of this study, the composition of the polyphenolic complex of the herb P. fruticosus L. was established, including the glycosides of quercetin, kaempferol, rhamnetin, proacyanidins, ellagic acid and ellagotannins. A method has been developed for the quantitative determination of the sum of aglycones of grass polyphenols of P. fruticosus L., which is based on the principle of extraction with the simultaneous hydrolysis of glycosides of polyphenols to aglycones and their subsequent chromatographic determination by high performance liquid chromatography and quantification using absolute calibration. Keywords—P. fruticosa L. herb, RP HPLC, quercetin glycosides, kaempferol glycosides, rhamnetin glycosides, proacyanidins, elagic acid, ellagotannins

the purpose of the present study was a comparative study of the ability of some freons to extract the essential oil of the roots of C. longa L. and to provide optimal conditions for its analysis. The dried, crushed roots of C. longa L. were used as raw materials (country of origin Uganda). Methoxynonafluorobutane (Novec 7100), fluoroketone and nhexane were used as extractants. Method of gas-liquid chromatography with mass detection was used. The most efficient extractant for ar-tumerone and curlone, which proved to be methoxynonafluorobutane (Novec 7100). The results of the studies indicate the prospects of using freons as extracting agents for the extraction of essential oil from the roots of C. longa L. The prospect consists in a more effective extracting ability of organofluorine solvents than n-hexane to increase the yield of both, the essential oil itself and the most pharmacologically relevant root components of C. longa L. ar-tumerone and curlone. Keywords—C. longa L. roots, ar-tumerone, curlone, methoxynonafluorobutane, fluoroketone, n-hexane.

The aim of this study was to investigate the kinetics of baicalin hydrolysis in the process of its extraction from Scutellaria baicalensis Georgi roots.Materials and methods. For the studies, Scutellaria baicalensis Georgi roots with a particle range of 0.1–0.5 mm were used. The method of extraction was a simple maceration during a specified period of time, the ratio of plant raw material : extractant was 1:10 w/v at the temperature of 24±1°С. Baicalin and baicalein contents were analyzed by reverse phase high performance liquid chromatography (RP HPLC) at the analytical wavelength of 275 nm. The extractant was a water solution of ethanol 26, 43, 59, 72, 81, 97±1% v/v. The time of the extraction was from 1 to 24 hours.Results. The experimental points of dependency of baicalin concentration in the extract on the time of extraction for ethanol solutions with a concentration of 43 and 72% v/v are closely approximated by a linear equation in coordinates lnC=f(t). The value of determination coefficient is more than R²˃0,99. Half lifetime for baicalin has been calculated: for ethanol with the concentration of 43% v/v it is 4.3±0.7 hours, and for ethanol with the concentration of 72% v/v it is 42.3±1.8 hours.Conclusion. Baicalin hydrolysis kinetics in the process of its extraction from Scutellaria baicalensis Georgi roots with 43 and 72% v/v ethanol concentration. has been studied. It has been established that the process of baicalin hydrolysis is well described by the first order kinetic equation. The constants of baicalin hydrolysis during its extraction from Scutelaria baicalensis roots with ethanol having different concentrations have been calculated. Recommendations on technology optimization for baicalin or baicalein extraction from Scutellaria baicalensis Georgi roots have been given.

The aim of this study is to confirm the adequacy of the proposed hypothesis, which explains and quantitatively describes the distribution of biologically active substances (BAS) within the extraction system consisting of Helichrysum arenarium flowers and the solvent using a regressive analysis for the theoretically predicted coordinates.Materials and methods. For this research, milled officinal flowers of Helichrysum arenarium (Helichrysum arenarium L. flores) were used. The analysis of the extractions was carried out by RP HPLC method. Isosalipurposide, salipurposide, and chlorogenic acid of >98.0% purity were used as reference substances. The analytical wavelengths were 370, 290, and 325 nm. Results. The obtained experimental data are well-approximated by regressive linear equations in the theoretically predicted coordinates 1/C=f(V) and ln(b/a)=f(1/T). Wherein, the coefficient of determination of regressive equations was R2>0.998, which indicates functional dependence between the studied parameters and confirms the adequacy of the developed mathematical model. The experimental work identified the necessity of implementation of additional constant values into the mathematical model.Conclusion. A new hypothesis was proposed to explain and quantitatively describe the distribution of BAS in the extraction system of Helichrysum arenarium flowers and 80% ethanol. With this working hypothesis, mathematical models were developed and their adequacy was proved using a regressive analysis in the theoretically predicted coordinates. The results obtained could not deny that a mechanism of BAS distribution between the phases is explained and described by the classic Boltzmann distribution for discrete values of molecular energy (or quantum distribution according to Fermi and Dirac).

Objective: The aim of this article was to test the hypothesis about the possibility for extraction of essential oil components from the plant raw material (PRM) by the example of Pimpinella anisum L. fruits with two different perfluoro organic solvents, Novec 1230 and Novec 7100. Methods: For the studies, we used Pimpinella anisum L. fruits ground to obtain particle size between 0.1 and 0.5 mm. The study of qualitative and quantitative content of the extracts was carried out by the following method: simple maceration, PRM: extractant ratio 1:10 w/v; and circulation method for extraction using Soxhlet extractor, PRM: extractant ratio 1:5 w/v. Qualitative analysis of extracts was carried out by gas chromatography-mass spectrometry (GC-MS) method. Quantitative analysis of trans-anethole in the extracts was carried out by reverse phase high-performance liquid chromatography (RP HPLC) method. Results: The largest yield of anethole was observed for n-hexane, 84±4 %, and for Novec 7100, 81±4 %. In case of Novec 1230 solvent, the yield of anethole was noted to be three times less, 32±2 %. The yield of anethole under the conditions of solvents (Novec 1230 and Novec 7100) circulation method was up to 94±5 % within two hours. Conclusion: Adequacy of the working hypothesis about the possibility for extraction of essential oil components from Pimpinella anisum L. fruits by two different perfluoro organic solvents, Novec 1230 and Novec 7100, was verified experimentally. In the context of the solvents tested, it was found that the level of fluorination of the solvent’s molecule had a great influence on its extraction properties in regard to anethole and vegetable oil.

Glutathione (γ-L-glutamyl-L-cysteinylglycine) is the most important low molecular weight intracellular thiol tripeptide consisting of three amino acids – glycine, cysteine and glutamic acid. In Russian pharmacopoeia there is no regulatory documentation for glutathione, therefore, the development of a pharmacopoeial item for the specified substance is a relevant problem.The aim of the article is the development of methods for determining foreign specific impurities in glutathione.Materials and methods. The substance of glutathione reduced (CAS 70-18-8, EC 2007254, Applichem, Germany) containing impurities, and a standard sample of reduced glutathione (Sigma Aldrich, Japan) were used as the objects of the study. The analysis was carried out by using a high-performance liquid chromatography method in the reverse phase version and a thin layer chromatography method. The chromatography using RP HPLC was performed after preliminary derivatization of glutathione and its specific impurities with dancil chloride. Specific impurities in glutathione are dipeptides and amino acids. Therefore, they, like glutathione, can react with dancil chloride. Dancil derivatives are formed, and they can be determined by chromatographic separation.Results. As a result of chromatography by the method of RP HPLC of derivatized dancil chloride glutathione it has been established that this reaction makes it possible to detect impurities in it. Glutathione derivatives are well separated by chromatography by implementing the method of RP HPLC and have different absorption maxima. The glutathione derivative had an absorption maximum at λmax=284 nm. The derivatives belonging to specific glutathione impurities absorb at λmax=288 nm and λmax=296 nm. The data obtained using RP HPLC were confirmed by TLC in the isopropanol-water (2:1) system. Three components were found out, one of which corresponds to glutathione, while two others are impurities.Conclusion. Methods for determining impurities in the glutathione substance using RP HPLC methods with preliminary derivatization with dancil chloride and TLC with ninhydrin detection have been worked out. A comparative analysis of the data obtained makes it possible to state that the OF-HPLC method with pre-column derivatization is more reliable, since it is more sensitive to impurities, and also makes it possible to study the UV profiles of impurity components better than the TLC method. Therefore, for the detection of impurities in the substance of glutathione, it is more preferable to use RP-HPLC with pre-column derivatization. The results of this study can be recommended for inclusion in the regulatory documentation on the substance of glutathione in the section “Impurities”.

Aim of this study is the theoretical justification and experimental confirmation of new approach to description of equilibrium state in extraction system vegetal raw materialextractant. In framework of this study the milled raw material was used containing the particles sized 0,1-0,5 mm: roots and rhyzomes of Glycyrrhiza glabra or uralensis, leaves of Uvaurzi, leaves of Eucalyptus viminalis, flowers of Helichrysum arenarium, flowers of Calendula officinalis and etc. The process of equilibrium establishment was studied at temperature levels such as 4, 20, 40 and 60±1oС, wherein the method of simple maceration was implemented for 24 hours of decoction. Interphase distribution of BAS was studied using the proportion of raw material weight / extractant volume as 1:5, 1:10, 1:20, 1:40. The watery ethanol solution of 70 or 80±1 % vol. was chosen as an extractant for the purpose of this study. Assay test of BAS in raw material was carried out with the help of RP HPLC instrumentation method. Experimental points for all the studied BAS from various types of vegetal raw materials are well-approximated by regression lines built up in theoretically predicted coordinates, which is confirmed by the high value of determination coefficient R2≥0.99. The new approach has been proposed to description of equilibrium state in extraction system vegetal raw material-extractant, which is based on the promotion of hypothesis, according to which the mechanism of interphase molecular distribution of BAS in extraction system is explained and described by classic Boltzmann distribution for discrete values of molecular energy (or Fermi-Dirac quantum distribution). The developed and experimentally confirmed mathematical model was based on the promoted hypothesis, and it successfully described the relation of the main parameters of extraction system, which allows to develop the rational technology of tincture or extracts manufacturing. Keywords—extraction, vegetal raw material, biologically active substances, equilibrium, Boltzmann distribution for discrete values of molecular energy, Fermi-Dirac quantum distribution.

The article presents the results related to the study of distribution of biologically active substances from the plant raw material between solid and liquid phases. The aim of this study is to develop theoretical bases of the extraction process in the equilibrium state by the example of study and modeling of the distribution process of biologically active substances from Eucalyptus viminalis leaves. In these studies, we used ground plant raw material of E. viminalis leaves with particle fraction of 0.1–0.5 mm; and ethanol with concentration 80% ±1% v/v was used as an extractant. Qualitative and quantitative analyses were carried out by reversed phase high-performance liquid chromatography with rutin, chlorogenic acid, and euglobal standards equivalent to spissum extract of chlorophyllipt of the State Pharmacopoeia of Ukraine. A hypothesis has been suggested that Henry's adsorption law and the law of conservation of matter play a fundamental role in this process. The experimental data are described well by the suggested equation with high value of determination coefficient R2 =0.99. At the same time, F-test and the significance of coefficients in equations satisfy the statistic condition, which means that the current hypothesis about the adsorption mechanism of distribution of biologically active substances in the extraction system is not refuted. The results of these studies demonstrate good agreement of experimental data and theoretical model based on Henry's adsorption law and mass balance. The numerical values of constants in the model suggested have been calculated.

The aim of this work is phytochemical studies and standardization of Scabiosa succisa L. By reverse phase high performance liquid chromatography of ethanol extract substances belonging to polyphenolic compounds were found, among which hydroxycinnamic acids predominate. Chlorogenic acid and flavonoid glycosides of apigenin and luteolin were identified. Microscopic analysis revealed the main diagnostic features of the Scabiosa succisa L. herb. As a result of research component composition of Scabiosa succisa L. polyphenols was partially installed, which substantiates its anti-inflammatory properties, and microdiagnostics signs were discovered, which can determine the plant authenticity. Keywords—Scabiosa succisa L., Caprifoliaceae, standardization, high-performance liquid chromatography, microscopy, phenolic compounds

The aim of the article was to study and model solvent effects on phytocompounds' extraction from Glycyrrhizae radix for substantiation of rational choice of the extractant in the technology of drugs obtained from this type of plant raw material. The process of extraction was carried out by simple maceration for 24 h at temperature 25°C ± 1°C and extractant/plant raw material ratio 5:1 (v/w). The content of hydro-ethanolic extracts based on ethanol solutions with concentration of 22, 41, 50, 71, 82, and 96% ± 1% v/v and some other solvents was studied by reversed phase high-performance liquid chromatography. The optimal range of solvent concentration for simultaneous extraction of chalcones and glycyrrhizic acid derivatives from Glycyrrhizae radix was determined. It was found that dielectric constant of the solvent plays a key role in the distribution process of phytocompounds between the phases; there is a certain range of dielectric constant values of the solvent-water solution, within which maximum phytocompound concentration in the extract can be observed; the dependency between phytocompound concentration in the extract and dielectric constant of the solvent-water solution can be described by equation lnC = a + b/ε+d/ε2.

Glutathione (γ-L-glutamyl-L-cysteinyl glycine) is the most important low molecular weight intracellular thiol tripeptide synthesized by almost all animal cells. Its role is to provide a number of important biological functions. Due to the sulfhydryl group of cysteine, it participates in the reactions of restoration and conjugation. Using these reactions, peroxides and many xenobiotic compounds are removed.[1] The reactions of peroxide elimination take place due to the fact that glutathione serves as the co-factor of glutathione peroxidase enzyme, while preventing the oxidation of free thiol groups of important proteins, including enzymes, the peroxidation of phospholipids in cell membranes is reduced.[2] The removal of xenobiotic compounds is carried out by direct conjugation with glutathione and the subsequent secretion of the adduct from the cell.[3] Thus, this molecule forms the basis of intracellular redox status, thereby protecting cells from active forms of oxygen.[4]

In work research results of the polyphenols chemical composition of Laurus nobilis L leaves are presented. Bay laurel is a known food plant having also wide range of important pharmacological properties such as: hypoglycemic, anti-inflammatory, antiepileptic and antioxidant. Using a method of a highly effective liquid chromatography in the gradient mode of elution, the structure of flavonoids of a plant which includes 12 components which are glycosides of quercetine and kaempferol is established. The technique of quantitative definition of flavonoids in raw materials of bay laurel in terms of hyperoside, consisting in application of a method of absolute graduation is developed. The maintenance of hyperoside in raw materials made 0.211%+/- 0.01.