Pub Date : 2022-04-09DOI: 10.32362/2410-6593-2022-17-1-39-49
A. D. Askretkov, D. Shatalov, N. Orlova, D. Zybin, V. V. Nikolaeva, A. Klishin, E. S. Tuzova, D. Minenkov, S. Kedik, Y. Seregin
Objectives. Polysorbate 80 (PS80) quantification in biopharmaceutical products has always been challenging owing to its minute content, absorption to the protein backbone, lack of specific chromophoric PS80 groups, and heterogenic nature. This work is aimed at developing an express method for PS80 analysis in biopharmaceutical products using hydrolysis and subsequent highperformance liquid chromatography analysis with ultraviolet detection that does not consume substantial amounts of sample (≥35 μL).Methods. Five therapeutic protein formulations were chosen as model proteins. Alkaline hydrolysis formulation was applied, without protein precipitation and with a range of precipitation techniques to remove protein from the test solution and hydrolyze PS80, to free fatty acids. The obtained hydrolysate was analyzed using reverse-phase high-performance liquid chromatography.Results. As a result of the high protein content of monoclonal antibody formulations, preliminary protein removal was required, which was achieved by precipitation with organic solvents. A specific precipitant ethanol–isopropanol mixture (1:1 volumetric ratio) was developed to efficiently remove antibodies while keeping PS80 in the solution. The PS80 quantification method was developed for monoclonal antibody drugs. For three monoclonal antibody drug products (adalimumab, infliximab, and eculizumab), method validation was performed according to the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use, the United States Pharmacopeia, and the State Pharmacopeia of the Russian Federation guidelines.Conclusions. The optimal assay conditions for each group of recombinant monoclonal antibody substances were chosen. Protein precipitation with ethanol or ethanol–isopropanol mixtures before hydrolysis was introduced, allowing for a substantial reduction of sample to 35 μL or even less if PS80 content is higher than 0.05 mg/mL. Accelerated hydrolysis (90 min) is preferable to slow hydrolysis (4–18 h). Method validation for protein products such as adalimumab, infliximab, and eculizumab was demonstrated for the first time. Both methods were validated for each drug product. The coefficients of variation for method specificity and high precision were ≤6.0% for 3 analyses. The accuracy of the methods ranged from 96% to 109% for all of the tested drug products.
{"title":"Quantitation of polysorbate 80 in recombinant protein formulation using high-performance liquid chromatography","authors":"A. D. Askretkov, D. Shatalov, N. Orlova, D. Zybin, V. V. Nikolaeva, A. Klishin, E. S. Tuzova, D. Minenkov, S. Kedik, Y. Seregin","doi":"10.32362/2410-6593-2022-17-1-39-49","DOIUrl":"https://doi.org/10.32362/2410-6593-2022-17-1-39-49","url":null,"abstract":"Objectives. Polysorbate 80 (PS80) quantification in biopharmaceutical products has always been challenging owing to its minute content, absorption to the protein backbone, lack of specific chromophoric PS80 groups, and heterogenic nature. This work is aimed at developing an express method for PS80 analysis in biopharmaceutical products using hydrolysis and subsequent highperformance liquid chromatography analysis with ultraviolet detection that does not consume substantial amounts of sample (≥35 μL).Methods. Five therapeutic protein formulations were chosen as model proteins. Alkaline hydrolysis formulation was applied, without protein precipitation and with a range of precipitation techniques to remove protein from the test solution and hydrolyze PS80, to free fatty acids. The obtained hydrolysate was analyzed using reverse-phase high-performance liquid chromatography.Results. As a result of the high protein content of monoclonal antibody formulations, preliminary protein removal was required, which was achieved by precipitation with organic solvents. A specific precipitant ethanol–isopropanol mixture (1:1 volumetric ratio) was developed to efficiently remove antibodies while keeping PS80 in the solution. The PS80 quantification method was developed for monoclonal antibody drugs. For three monoclonal antibody drug products (adalimumab, infliximab, and eculizumab), method validation was performed according to the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use, the United States Pharmacopeia, and the State Pharmacopeia of the Russian Federation guidelines.Conclusions. The optimal assay conditions for each group of recombinant monoclonal antibody substances were chosen. Protein precipitation with ethanol or ethanol–isopropanol mixtures before hydrolysis was introduced, allowing for a substantial reduction of sample to 35 μL or even less if PS80 content is higher than 0.05 mg/mL. Accelerated hydrolysis (90 min) is preferable to slow hydrolysis (4–18 h). Method validation for protein products such as adalimumab, infliximab, and eculizumab was demonstrated for the first time. Both methods were validated for each drug product. The coefficients of variation for method specificity and high precision were ≤6.0% for 3 analyses. The accuracy of the methods ranged from 96% to 109% for all of the tested drug products.","PeriodicalId":12215,"journal":{"name":"Fine Chemical Technologies","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85392258","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-04-09DOI: 10.32362/2410-6593-2022-17-1-18-29
B. Y. Malyshkin, I. P. Semenov, D. Sazonov, I. O. Putenikhin, E. Y. Semenikhina
Objectives. This study aimed to create a method for purifying benzene fractions using a modified natural clay-based adsorbent, which removes both unsaturated and sulfur compounds from benzene or benzene fractions obtained by pyrolysis of petrochemical raw materials.Methods. Chemical and chromatographic methods were used to conduct flow analyses before and after purification. Testing was conducted under conditions that were similar to industrial conditions. The functional characteristics of the adsorbents (moisture resistance and bulk density) were evaluated by strength tests after wetting with water and by weighing batches of the adsorbents fixed in volume.Results. Three clay-based adsorbent samples were developed, and tested under laboratory conditions. Thereafter, the CS-Sorb-6890 (M) sample—an Ni-modified adsorbent—was tested under experimental industrial conditions. A positive effect was obtained, and a method for purifying benzene fractions (benzene) from sulfur compounds and olefins is proposed.Conclusions. The obtained results revealed that the modified active clay-based adsorbent successfully removed sulfur and unsaturated compounds from the benzene fraction simultaneously. The total sulfur concentration was reduced to 0.00004 wt % after 90% of it was removed. The conversion of the unsaturated compounds reached 100%.
{"title":"Purification of benzene fractions and benzene from unsaturated and sulfur compounds using a novel modified natural clay-based adsorbent","authors":"B. Y. Malyshkin, I. P. Semenov, D. Sazonov, I. O. Putenikhin, E. Y. Semenikhina","doi":"10.32362/2410-6593-2022-17-1-18-29","DOIUrl":"https://doi.org/10.32362/2410-6593-2022-17-1-18-29","url":null,"abstract":"Objectives. This study aimed to create a method for purifying benzene fractions using a modified natural clay-based adsorbent, which removes both unsaturated and sulfur compounds from benzene or benzene fractions obtained by pyrolysis of petrochemical raw materials.Methods. Chemical and chromatographic methods were used to conduct flow analyses before and after purification. Testing was conducted under conditions that were similar to industrial conditions. The functional characteristics of the adsorbents (moisture resistance and bulk density) were evaluated by strength tests after wetting with water and by weighing batches of the adsorbents fixed in volume.Results. Three clay-based adsorbent samples were developed, and tested under laboratory conditions. Thereafter, the CS-Sorb-6890 (M) sample—an Ni-modified adsorbent—was tested under experimental industrial conditions. A positive effect was obtained, and a method for purifying benzene fractions (benzene) from sulfur compounds and olefins is proposed.Conclusions. The obtained results revealed that the modified active clay-based adsorbent successfully removed sulfur and unsaturated compounds from the benzene fraction simultaneously. The total sulfur concentration was reduced to 0.00004 wt % after 90% of it was removed. The conversion of the unsaturated compounds reached 100%.","PeriodicalId":12215,"journal":{"name":"Fine Chemical Technologies","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81196081","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-27DOI: 10.32362/2410-6593-2021-16-6-512-525
D. Aleksandrova, T. B. Melamed, E. Baberkina, A. A. Fenin, E. S. Osinova, А. Kovalenko, R. Yakushin, Y. Shaltaeva, V. Belyakov, D. I. Zykova
Objectives. To determine the ion mobility of N-methylimidazole, establish the structure of ions corresponding to characteristic signals, and determine the detection limit of N-methylimidazole on the ion-drift detector Kerber.Methods. Ion mobility spectrometry was used to study the ionization processes. The enthalpies of the reactions of monomer and dimer ions were calculated in the ORCA 4.1.1 software by the B3LYP density functional method with a set of basic functions 6-31G (d, p).Results. The drift time and ion mobility values of N-methylimidazole were determined. A method for mathematical processing of spectra and a program for its implementation was developed. The changing peculiarities of the ion mobility spectrum during measurement at a given time were studied. According to the interpretation of the spectrum signals, the structure of the generated ions was proposed, and the enthalpies of ion formation were determined.Conclusions. The characteristic signal of the N-methylimidazole ion protonated at the nitrogen atom of the pyridine type was revealed. It was found that two signals in the ion mobility spectra of N-methylimidazole correspond to the presence of the monomer and dimer ions. The detection limit of N-methylimidazole on the ion-drift detector Kerber was determined, amounting to 3 pg.
{"title":"Ion mobility spectrometry of N-methylimidazole and possibilities of its determination","authors":"D. Aleksandrova, T. B. Melamed, E. Baberkina, A. A. Fenin, E. S. Osinova, А. Kovalenko, R. Yakushin, Y. Shaltaeva, V. Belyakov, D. I. Zykova","doi":"10.32362/2410-6593-2021-16-6-512-525","DOIUrl":"https://doi.org/10.32362/2410-6593-2021-16-6-512-525","url":null,"abstract":"Objectives. To determine the ion mobility of N-methylimidazole, establish the structure of ions corresponding to characteristic signals, and determine the detection limit of N-methylimidazole on the ion-drift detector Kerber.Methods. Ion mobility spectrometry was used to study the ionization processes. The enthalpies of the reactions of monomer and dimer ions were calculated in the ORCA 4.1.1 software by the B3LYP density functional method with a set of basic functions 6-31G (d, p).Results. The drift time and ion mobility values of N-methylimidazole were determined. A method for mathematical processing of spectra and a program for its implementation was developed. The changing peculiarities of the ion mobility spectrum during measurement at a given time were studied. According to the interpretation of the spectrum signals, the structure of the generated ions was proposed, and the enthalpies of ion formation were determined.Conclusions. The characteristic signal of the N-methylimidazole ion protonated at the nitrogen atom of the pyridine type was revealed. It was found that two signals in the ion mobility spectra of N-methylimidazole correspond to the presence of the monomer and dimer ions. The detection limit of N-methylimidazole on the ion-drift detector Kerber was determined, amounting to 3 pg.","PeriodicalId":12215,"journal":{"name":"Fine Chemical Technologies","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89333149","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-27DOI: 10.32362/2410-6593-2021-16-6-502-511
E. Nikishina
Objectives. The study aimed to develop new methods for the synthesis of cobalt ferrite (CoFe2O4), which is a precursor for the synthesis of CoFe2O4-based functional materials, as well as to study the physicochemical properties of the obtained phases.Methods. Two methods were used for the synthesis of CoFe2O4: (1) heterophase interaction of hydrated iron oxide with cobalt(II, III) oxide and (2) heterophase interaction of hydrated iron oxide with an aqueous solution of cobalt(II) sulfate (CCo = 0.147 mol/L, solid/liquid = 1:43). In both cases, the precursor was hydrated iron oxide (Fe2O3, 84.4 wt %), which was obtained by the heterophase interaction of iron(III) chloride with a concentrated ammonia solution (6.0–9.5 mol/L). The resulting intermediate products were subjected to thermal treatment at 750 °C (synthesis 1) and at 900 °C (synthesis 2) for 10–30 h in increments of 10 h. The synthesized phases and products of their thermolysis were studied by differential thermal analysis and differential thermogravimetry (DTA–DTG), X-ray diffraction analysis (XRDA), and granulometry.Results. The hydrated iron oxide sample remained amorphous even up to the crystallization temperature of 445 °C, which corresponds to the exothermic effect on the DTA curve. Further heating led to the α-modification of iron(III) oxide of the hexagonal system (a = b = 5.037 ± 0.002 Å; c = 13.74 ± 0.01 Å), which has an average particle size of 1.1 μm. XRDA results showed that a synthesis temperature of 750 °C (synthesis 1) and a heat treatment duration of 30 h were sufficient for the formation of a single-phase cobalt ferrite (a = 8.388 ± 0.002 Å) with an average particle diameter of 1.9 μm. For synthesis 2, a higher temperature of 900 °C was used because sample weight loss (about 12.5%) was observed in the temperature range of 720–810 °C based on the DTA results, which was due to the removal of SO2 and SO3. Moreover, when synthesis temperature and duration were at 900 °C and 30 h, respectively, CoFe2O4 with a = 8.389 ± 0.002 Å was formed. The results of the granulometric analysis showed that particles of different diameters were formed. The smallest particle size (1.5 μm) of cobalt ferrite was obtained by the heterophase interaction of hydrated iron(III) oxide (Fe2O3, 84.4 wt %) with an aqueous solution of cobalt sulfate with CCo = 0.147 mol/L. Conclusions. Depending on the method used for the synthesis of cobalt ferrite, particles of different diameters are formed. The smallest particle size of cobalt ferrite was obtained from the heterophase interaction of hydrated iron(III) oxide with an aqueous solution of cobalt(II) sulfate.
{"title":"Heterophase synthesis of ferrite cobalt Elena E. Nikishina","authors":"E. Nikishina","doi":"10.32362/2410-6593-2021-16-6-502-511","DOIUrl":"https://doi.org/10.32362/2410-6593-2021-16-6-502-511","url":null,"abstract":"Objectives. The study aimed to develop new methods for the synthesis of cobalt ferrite (CoFe2O4), which is a precursor for the synthesis of CoFe2O4-based functional materials, as well as to study the physicochemical properties of the obtained phases.Methods. Two methods were used for the synthesis of CoFe2O4: (1) heterophase interaction of hydrated iron oxide with cobalt(II, III) oxide and (2) heterophase interaction of hydrated iron oxide with an aqueous solution of cobalt(II) sulfate (CCo = 0.147 mol/L, solid/liquid = 1:43). In both cases, the precursor was hydrated iron oxide (Fe2O3, 84.4 wt %), which was obtained by the heterophase interaction of iron(III) chloride with a concentrated ammonia solution (6.0–9.5 mol/L). The resulting intermediate products were subjected to thermal treatment at 750 °C (synthesis 1) and at 900 °C (synthesis 2) for 10–30 h in increments of 10 h. The synthesized phases and products of their thermolysis were studied by differential thermal analysis and differential thermogravimetry (DTA–DTG), X-ray diffraction analysis (XRDA), and granulometry.Results. The hydrated iron oxide sample remained amorphous even up to the crystallization temperature of 445 °C, which corresponds to the exothermic effect on the DTA curve. Further heating led to the α-modification of iron(III) oxide of the hexagonal system (a = b = 5.037 ± 0.002 Å; c = 13.74 ± 0.01 Å), which has an average particle size of 1.1 μm. XRDA results showed that a synthesis temperature of 750 °C (synthesis 1) and a heat treatment duration of 30 h were sufficient for the formation of a single-phase cobalt ferrite (a = 8.388 ± 0.002 Å) with an average particle diameter of 1.9 μm. For synthesis 2, a higher temperature of 900 °C was used because sample weight loss (about 12.5%) was observed in the temperature range of 720–810 °C based on the DTA results, which was due to the removal of SO2 and SO3. Moreover, when synthesis temperature and duration were at 900 °C and 30 h, respectively, CoFe2O4 with a = 8.389 ± 0.002 Å was formed. The results of the granulometric analysis showed that particles of different diameters were formed. The smallest particle size (1.5 μm) of cobalt ferrite was obtained by the heterophase interaction of hydrated iron(III) oxide (Fe2O3, 84.4 wt %) with an aqueous solution of cobalt sulfate with CCo = 0.147 mol/L. Conclusions. Depending on the method used for the synthesis of cobalt ferrite, particles of different diameters are formed. The smallest particle size of cobalt ferrite was obtained from the heterophase interaction of hydrated iron(III) oxide with an aqueous solution of cobalt(II) sulfate.","PeriodicalId":12215,"journal":{"name":"Fine Chemical Technologies","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78430974","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-27DOI: 10.32362/2410-6593-2021-16-6-526-540
E. Kartashov
Objectives. This study mathematically describes the mutual influence of micro- and macrostages of the process of destruction of polymer materials and determines its main parameters and limiting characteristics. In addition, a relationship is established between molecular constants characterizing the structure of a material and those characterizing its macroscopic characteristics of strength. Finally, theoretical representations of the thermokinetics of the process of thermal destruction of polymer fibers from the standpoint of the kinetic thermofluctuation concept are developed, which makes it possible to predict the thermal durability of a sample under thermal loading.Methods. The structural–kinetic thermofluctuation theory was used to describe the initial stages of the fracture process and to derive a generalized formula for the rate of crack growth. The mathematical theory of cracks is used to describe the thermally stressed state of a material in the vicinity of an internal circular crack under mechanical and thermal loadings of the sample.Results. A theoretical formula for the full isotherm of durability in the range of mechanical stresses from safe to critical, as well as a theoretical relationship for the time dependence of the strength of polymer fibers under purely thermal loading in the full range of heat loads from safe to critical and at the stage of nonthermal crack growth, is given. The main parameters and limiting characteristics of durability under thermal loading are also indicated.Conclusions. A generalized structural–kinetic theory of the fracture of polymer fibers under purely thermal action on cracked specimens is presented. The developed theory combines three independent approaches: structural–kinetic (thermofluctuation theory), mechanical, and thermodynamic. The obtained theoretical relations are of practical interest for the development of methods for localization, intensification, and control of the crack growth kinetics.
{"title":"Thermal destruction of polymeric fibers in the theory of temporary dependence of strength","authors":"E. Kartashov","doi":"10.32362/2410-6593-2021-16-6-526-540","DOIUrl":"https://doi.org/10.32362/2410-6593-2021-16-6-526-540","url":null,"abstract":"Objectives. This study mathematically describes the mutual influence of micro- and macrostages of the process of destruction of polymer materials and determines its main parameters and limiting characteristics. In addition, a relationship is established between molecular constants characterizing the structure of a material and those characterizing its macroscopic characteristics of strength. Finally, theoretical representations of the thermokinetics of the process of thermal destruction of polymer fibers from the standpoint of the kinetic thermofluctuation concept are developed, which makes it possible to predict the thermal durability of a sample under thermal loading.Methods. The structural–kinetic thermofluctuation theory was used to describe the initial stages of the fracture process and to derive a generalized formula for the rate of crack growth. The mathematical theory of cracks is used to describe the thermally stressed state of a material in the vicinity of an internal circular crack under mechanical and thermal loadings of the sample.Results. A theoretical formula for the full isotherm of durability in the range of mechanical stresses from safe to critical, as well as a theoretical relationship for the time dependence of the strength of polymer fibers under purely thermal loading in the full range of heat loads from safe to critical and at the stage of nonthermal crack growth, is given. The main parameters and limiting characteristics of durability under thermal loading are also indicated.Conclusions. A generalized structural–kinetic theory of the fracture of polymer fibers under purely thermal action on cracked specimens is presented. The developed theory combines three independent approaches: structural–kinetic (thermofluctuation theory), mechanical, and thermodynamic. The obtained theoretical relations are of practical interest for the development of methods for localization, intensification, and control of the crack growth kinetics.","PeriodicalId":12215,"journal":{"name":"Fine Chemical Technologies","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78472587","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-27DOI: 10.32362/2410-6593-2021-16-6-476-489
E. Pashkov, E. Faizuloev, E. Korchevaya, A. Rtishchev, B. Cherepovich, А. V. Sidorov, A. Poddubikov, Е. P. Bystritskaya, Y. Dronina, A. Bykov, O. Svitich, V. Zverev
Objectives. To evaluate the effect of cellular genes FLT4, Nup98, and Nup205 on the reproduction of the influenza A virus in A549 human lung cancer cell line.Methods. The work was carried out using the equipment of the center for collective use of the I.I. Mechnikov Research Institute of Vaccines and Sera (Russia). The virus-containing fluid was collected within three days from the moment of transfection and infection and the intensity of viral reproduction was assessed by viral titration and hemagglutination reaction. The viral RNA concentration was determined by real-time reverse-transcription polymerase chain reaction (RT-PCR). To calculate statistically significant differences between groups, the nonparametric Mann–Whitney test was used.Results. In cells treated with small interfering RNAs (siRNAs) targeted at FLT4, Nup98, and Nup205 genes, a significant decrease in their expression and indicators of viral reproduction (virus titer, hemagglutinating activity, viral RNA concentration) was observed at a multiplicity of infection (MOI) = 0.1. Additionally, it was found that a decrease in the expression of target genes using siRNA does not lead to a significant decrease in cell survival. The viral titer in cells treated with siRNA FLT4.2, Nup98.1, and Nup205 on the first day was lower by an average of 1.0 lg, and on the second and third days, by 2.2–2.3 lg, compared to cells treated with nonspecific siRNA. During real-time RT-PCR, a significant decrease in the concentration of viral RNA was observed with siRNA Nup98.1 (up to 190 times) and Nup205 (up to 30 times) on the first day, 26 and 29 times on the second day, and 6 and 30 times on the third day, respectively. For FLT4.2 siRNA, the number of viral RNA copies decreased by 23, 18, and 16 times on the first, second, and third days. Similar results were obtained when determining the hemagglutinating activity of the virus. The hemagglutinating activity on the third day most strongly decreased in cells treated with siRNA Nup205 and FLT4.2 (16 times). In cells treated with siRNA FLT4.1, Nup98.1, and Nup98.2, hemagglutinating activity decreased by 8 times.Conclusions. In the present study, three cellular genes (FLT4, Nup98, and Nup205) were identified—the decrease in the expression of which effectively suppresses viral reproduction— and the original siRNA sequences were obtained. The results obtained are important for creating therapeutic and prophylactic medication, whose action is based on the RNA interference mechanism.
{"title":"Knockdown of FLT4, Nup98, and Nup205 cellular genes as a suppressor for the viral activity of Influenza A/WSN/33 (H1N1) in A549 cell culture","authors":"E. Pashkov, E. Faizuloev, E. Korchevaya, A. Rtishchev, B. Cherepovich, А. V. Sidorov, A. Poddubikov, Е. P. Bystritskaya, Y. Dronina, A. Bykov, O. Svitich, V. Zverev","doi":"10.32362/2410-6593-2021-16-6-476-489","DOIUrl":"https://doi.org/10.32362/2410-6593-2021-16-6-476-489","url":null,"abstract":"Objectives. To evaluate the effect of cellular genes FLT4, Nup98, and Nup205 on the reproduction of the influenza A virus in A549 human lung cancer cell line.Methods. The work was carried out using the equipment of the center for collective use of the I.I. Mechnikov Research Institute of Vaccines and Sera (Russia). The virus-containing fluid was collected within three days from the moment of transfection and infection and the intensity of viral reproduction was assessed by viral titration and hemagglutination reaction. The viral RNA concentration was determined by real-time reverse-transcription polymerase chain reaction (RT-PCR). To calculate statistically significant differences between groups, the nonparametric Mann–Whitney test was used.Results. In cells treated with small interfering RNAs (siRNAs) targeted at FLT4, Nup98, and Nup205 genes, a significant decrease in their expression and indicators of viral reproduction (virus titer, hemagglutinating activity, viral RNA concentration) was observed at a multiplicity of infection (MOI) = 0.1. Additionally, it was found that a decrease in the expression of target genes using siRNA does not lead to a significant decrease in cell survival. The viral titer in cells treated with siRNA FLT4.2, Nup98.1, and Nup205 on the first day was lower by an average of 1.0 lg, and on the second and third days, by 2.2–2.3 lg, compared to cells treated with nonspecific siRNA. During real-time RT-PCR, a significant decrease in the concentration of viral RNA was observed with siRNA Nup98.1 (up to 190 times) and Nup205 (up to 30 times) on the first day, 26 and 29 times on the second day, and 6 and 30 times on the third day, respectively. For FLT4.2 siRNA, the number of viral RNA copies decreased by 23, 18, and 16 times on the first, second, and third days. Similar results were obtained when determining the hemagglutinating activity of the virus. The hemagglutinating activity on the third day most strongly decreased in cells treated with siRNA Nup205 and FLT4.2 (16 times). In cells treated with siRNA FLT4.1, Nup98.1, and Nup98.2, hemagglutinating activity decreased by 8 times.Conclusions. In the present study, three cellular genes (FLT4, Nup98, and Nup205) were identified—the decrease in the expression of which effectively suppresses viral reproduction— and the original siRNA sequences were obtained. The results obtained are important for creating therapeutic and prophylactic medication, whose action is based on the RNA interference mechanism.","PeriodicalId":12215,"journal":{"name":"Fine Chemical Technologies","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76594634","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-27DOI: 10.32362/2410-6593-2021-16-6-490-501
A. N. Stuzhuk, A. V. Shkolnikov, P. S. Gorbatov, I. Gritskova
Objectives. To create stable artificial polymer suspensions with a positive charge of particles based on polycarbonate and polymethyl methacrylate using cationic surfactants and organosilicon surfactants.Methods. The size of droplets and polymer suspension particles was determined by photon correlation spectroscopy (dynamic light scattering) using a Zetasizer NanoZS laser particle analyzer (Malvern, UK).Results. Domestic cationic surfactants Katamin-AB and Azol-129 were found to be capable of producing stable artificial polycarbonate and polymethyl methacrylate suspensions. Based on the polymer, the optimal surfactant concentration was 6 wt %. The effect of polymer concentration in solution on the stability and particle size of final polymer suspensions was shown. It was determined that the polymer concentration in the solution should not exceed 10%. When obtaining a highly dispersed suspension during dispersion, a higher concentration causes an increase in the viscosity of emulsions. As a result of a synergistic effect formation, we used mixtures of cationic surfactants (Katamin-AB/Azol-138 and Azol-129/Azol-138) to enhance the stability of the final polymer suspensions. The optimal surfactant ratio was 9:1. The total concentration of the mixture is 10 wt %, based on the polymer. Polymer suspensions were stabilized with each of 2:1 mixtures of cationic surfactants Katamin-AB and Azol-129 withan organosilicon surfactant U-851. The total mixture concentration was 9 wt %, based on the polymer.Conclusions. New methods of producing artificial polycarbonate and polymethyl methacrylate suspensions in the presence of domestically produced cationic surfactants, as well cationicorganosilicon surfactants mixtures, were proposed. The colloidal-chemical properties of the obtained polymer suspensions were considered. It was found that using a 2:1 mixture of cationic and organosilicon surfactants produces polymer suspensions that are stable during production and storage.
{"title":"Influence of emulgator nature on dispersity and stability of artificial polymer suspensions based on polycarbonate and polymethyl methacrylate","authors":"A. N. Stuzhuk, A. V. Shkolnikov, P. S. Gorbatov, I. Gritskova","doi":"10.32362/2410-6593-2021-16-6-490-501","DOIUrl":"https://doi.org/10.32362/2410-6593-2021-16-6-490-501","url":null,"abstract":"Objectives. To create stable artificial polymer suspensions with a positive charge of particles based on polycarbonate and polymethyl methacrylate using cationic surfactants and organosilicon surfactants.Methods. The size of droplets and polymer suspension particles was determined by photon correlation spectroscopy (dynamic light scattering) using a Zetasizer NanoZS laser particle analyzer (Malvern, UK).Results. Domestic cationic surfactants Katamin-AB and Azol-129 were found to be capable of producing stable artificial polycarbonate and polymethyl methacrylate suspensions. Based on the polymer, the optimal surfactant concentration was 6 wt %. The effect of polymer concentration in solution on the stability and particle size of final polymer suspensions was shown. It was determined that the polymer concentration in the solution should not exceed 10%. When obtaining a highly dispersed suspension during dispersion, a higher concentration causes an increase in the viscosity of emulsions. As a result of a synergistic effect formation, we used mixtures of cationic surfactants (Katamin-AB/Azol-138 and Azol-129/Azol-138) to enhance the stability of the final polymer suspensions. The optimal surfactant ratio was 9:1. The total concentration of the mixture is 10 wt %, based on the polymer. Polymer suspensions were stabilized with each of 2:1 mixtures of cationic surfactants Katamin-AB and Azol-129 withan organosilicon surfactant U-851. The total mixture concentration was 9 wt %, based on the polymer.Conclusions. New methods of producing artificial polycarbonate and polymethyl methacrylate suspensions in the presence of domestically produced cationic surfactants, as well cationicorganosilicon surfactants mixtures, were proposed. The colloidal-chemical properties of the obtained polymer suspensions were considered. It was found that using a 2:1 mixture of cationic and organosilicon surfactants produces polymer suspensions that are stable during production and storage.","PeriodicalId":12215,"journal":{"name":"Fine Chemical Technologies","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74870882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-26DOI: 10.32362/2410-6593-2021-16-6-457-464
A. V. Frolkova, V. G. Fertikova, E. Rytova, A. Frolkova
Objectives. The purpose of the paper is to compare the adequacy of mathematical models of vapor–liquid equilibrium (VLE) and their ability to reproduce the phase behavior of the ternary system benzene–cyclohexane–chlorobenzene using different experimental data sets to evaluate binary interaction parameters.Methods. The research methodologies were mathematical modeling of VLE in the Aspen Plus V.10.0 software package using activity coefficient models (Non-Random Two-Liquid (NRTL), Wilson) and the Universal quasichemical Functional-group Activity Coefficients (UNIFAC) group model, which allows for independent information. For the benzene–cyclohexane–chlorobenzene ternary system, the use of the NRTL equation is warranted because it provides a better description of the VLE experimental data.Results. The diagram construction of the constant volatility of cyclohexane relative to benzene lines revealed three topological structures. Only one of them can be considered reliable because it corresponds to the experimental data and coincides with the UNIFAC model diagram constructed based on independent UNIFAC model data. The results indicate that to study systems containing components with similar properties, it is necessary to improve the description quality of the available data sets (the relative error should not exceed 1.5%).Conclusions. The reproduction of the thermodynamic features of various manifolds in the composition simplex obtained by processing direct VLE data can be used to supplement the adequacy of the model. For the cyclohexane–benzene–chlorobenzene system, the best NRTL equation parameters are those regressed from the extensive experimental VLE data available in the literature for the ternary system as a whole.
目标。本文的目的是比较气液平衡(VLE)数学模型的充分性及其在用不同的实验数据集来评估二元相互作用参数时再现三元体系苯-环己烷-氯苯相行为的能力。研究方法是在Aspen Plus V.10.0软件包中使用活度系数模型(非随机双液(NRTL)， Wilson)和通用准化学官能团活度系数(UNIFAC)组模型对VLE进行数学建模，允许独立信息。对于苯-环己烷-氯苯三元体系，使用NRTL方程是有必要的，因为它能更好地描述VLE实验数据。环己烷相对于苯的恒挥发性谱图显示了三种拓扑结构。其中只有一个与实验数据相对应，且与基于独立UNIFAC模型数据构建的UNIFAC模型图吻合，可以认为是可靠的。结果表明，要研究含有相似性质成分的系统，必须提高可用数据集的描述质量(相对误差不超过1.5%)。通过直接VLE数据处理得到的组成单纯形中各种流形的热力学特征的再现可以用来补充模型的充分性。对于环己烷-苯-氯苯体系，最好的NRTL方程参数是根据文献中广泛的实验VLE数据对整个三元体系进行回归的。
{"title":"Evaluation of the adequacy of phase equilibria modeling based on various sets of experimental data","authors":"A. V. Frolkova, V. G. Fertikova, E. Rytova, A. Frolkova","doi":"10.32362/2410-6593-2021-16-6-457-464","DOIUrl":"https://doi.org/10.32362/2410-6593-2021-16-6-457-464","url":null,"abstract":"Objectives. The purpose of the paper is to compare the adequacy of mathematical models of vapor–liquid equilibrium (VLE) and their ability to reproduce the phase behavior of the ternary system benzene–cyclohexane–chlorobenzene using different experimental data sets to evaluate binary interaction parameters.Methods. The research methodologies were mathematical modeling of VLE in the Aspen Plus V.10.0 software package using activity coefficient models (Non-Random Two-Liquid (NRTL), Wilson) and the Universal quasichemical Functional-group Activity Coefficients (UNIFAC) group model, which allows for independent information. For the benzene–cyclohexane–chlorobenzene ternary system, the use of the NRTL equation is warranted because it provides a better description of the VLE experimental data.Results. The diagram construction of the constant volatility of cyclohexane relative to benzene lines revealed three topological structures. Only one of them can be considered reliable because it corresponds to the experimental data and coincides with the UNIFAC model diagram constructed based on independent UNIFAC model data. The results indicate that to study systems containing components with similar properties, it is necessary to improve the description quality of the available data sets (the relative error should not exceed 1.5%).Conclusions. The reproduction of the thermodynamic features of various manifolds in the composition simplex obtained by processing direct VLE data can be used to supplement the adequacy of the model. For the cyclohexane–benzene–chlorobenzene system, the best NRTL equation parameters are those regressed from the extensive experimental VLE data available in the literature for the ternary system as a whole.","PeriodicalId":12215,"journal":{"name":"Fine Chemical Technologies","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81160124","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-26DOI: 10.32362/2410-6593-2021-16-6-465-475
A. Ha
Objectives. The study aimed to analyze the current antiseptics and disinfectants, explore the possibility of synthesizing various antiseptics including oligohexamethylene guanidine hydrochloride (OHMG-HC) using microfluidic technology, and investigate the main synthesis parameters affecting the properties of the resulting product.Methods. This article presented a review of literature sources associated with investigations of antimicrobial resistance, the uses of agents based on polyhexamethylene guanidine hydrochloride, oligohexamethylene guanidine hydrochloride, and other salts, obained using modern synthesis technologies with microreactors.Results. The relevance of developing production technologies for the “OHMG-HC branched” substance was determined. The microfluidic method for the synthesis of polymers, and its application prospects for obtaining the target substance were compared with the existing methods. Advantages of the microfluidic method were indicated.Conclusions. Microreactor technologies allow for more accurate control of the conditions of the polycondensation reaction of the starting monomers and increase the yield and selectivity of the oligomers obtained, leading to an increase in the product purity and process efficiency, in contrast with other known methods. The use of microreactor technologies for the synthesis of branched oligohexamethylene guanidine hydrochloride products is a promising strategy.
{"title":"Ho Chi Minh City University of Technology (HCMUT); Vietnam National University Ho Chi Minh City (VNU-HCM)","authors":"A. Ha","doi":"10.32362/2410-6593-2021-16-6-465-475","DOIUrl":"https://doi.org/10.32362/2410-6593-2021-16-6-465-475","url":null,"abstract":"Objectives. The study aimed to analyze the current antiseptics and disinfectants, explore the possibility of synthesizing various antiseptics including oligohexamethylene guanidine hydrochloride (OHMG-HC) using microfluidic technology, and investigate the main synthesis parameters affecting the properties of the resulting product.Methods. This article presented a review of literature sources associated with investigations of antimicrobial resistance, the uses of agents based on polyhexamethylene guanidine hydrochloride, oligohexamethylene guanidine hydrochloride, and other salts, obained using modern synthesis technologies with microreactors.Results. The relevance of developing production technologies for the “OHMG-HC branched” substance was determined. The microfluidic method for the synthesis of polymers, and its application prospects for obtaining the target substance were compared with the existing methods. Advantages of the microfluidic method were indicated.Conclusions. Microreactor technologies allow for more accurate control of the conditions of the polycondensation reaction of the starting monomers and increase the yield and selectivity of the oligomers obtained, leading to an increase in the product purity and process efficiency, in contrast with other known methods. The use of microreactor technologies for the synthesis of branched oligohexamethylene guanidine hydrochloride products is a promising strategy.","PeriodicalId":12215,"journal":{"name":"Fine Chemical Technologies","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88369597","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-11-28DOI: 10.32362/2410-6593-2021-16-5-414-425
K. Rozhkov, E. Yagudaeva, S. Sizova, M. Lazov, E. V. Smirnova, V. Zubov, A. Ischenko
Objectives. This paper presents data on the development and study of the structural properties of iron-doped crystalline silicon (nc-Si/SiOx/Fe) nanoparticles obtained using the plasma-chemical method for application in magnetic resonance imaging diagnostics and treatment of oncological diseases. This work aimed to use a variety of analytical methods to study the structural properties of nc-Si/SiOx/Fe and their colloidal stabilization with citrate anions for in vivo applications.Methods. Silicon nanoparticles obtained via the plasma-chemical synthesis method were characterized by laser spark emission spectroscopy, atomic emission spectroscopy, Fouriertransform infrared spectroscopy, and X-ray photoelectron spectroscopy. The hydrodynamic diameter of the nanoparticles was estimated using dynamic light scattering. The toxicity of the nanoparticles was investigated using a colorimetric MTT test for the cell metabolic activity. Elemental iron with different Fe/Si atomic ratios was added to the feedstock during loading.Results. The particles were shown to have a large silicon core covered by a relatively thin layer of intermediate oxides (interface) and an amorphous oxide shell, which is silicon oxide with different oxidation states SiOx (0 ≤ x ≤ 2). The samples had an iron content of 0.8–1.8 at %. Colloidal solutions of the nanoparticles stabilized by citrate anions were obtained and characterized. According to the analysis of the cytotoxicity of the modified nanosilicon particles using monoclonal K562 human erythroleukemia cells, no toxicity was found for cells in culture at particle concentrations of up to 5 µg/mL.Conclusions. Since the obtained modified particles are nontoxic, they can be used in in vivo theranostic applications.
{"title":"Characterization of iron-doped crystalline silicon nanoparticles and their modification with citrate anions for in vivo applications","authors":"K. Rozhkov, E. Yagudaeva, S. Sizova, M. Lazov, E. V. Smirnova, V. Zubov, A. Ischenko","doi":"10.32362/2410-6593-2021-16-5-414-425","DOIUrl":"https://doi.org/10.32362/2410-6593-2021-16-5-414-425","url":null,"abstract":"Objectives. This paper presents data on the development and study of the structural properties of iron-doped crystalline silicon (nc-Si/SiOx/Fe) nanoparticles obtained using the plasma-chemical method for application in magnetic resonance imaging diagnostics and treatment of oncological diseases. This work aimed to use a variety of analytical methods to study the structural properties of nc-Si/SiOx/Fe and their colloidal stabilization with citrate anions for in vivo applications.Methods. Silicon nanoparticles obtained via the plasma-chemical synthesis method were characterized by laser spark emission spectroscopy, atomic emission spectroscopy, Fouriertransform infrared spectroscopy, and X-ray photoelectron spectroscopy. The hydrodynamic diameter of the nanoparticles was estimated using dynamic light scattering. The toxicity of the nanoparticles was investigated using a colorimetric MTT test for the cell metabolic activity. Elemental iron with different Fe/Si atomic ratios was added to the feedstock during loading.Results. The particles were shown to have a large silicon core covered by a relatively thin layer of intermediate oxides (interface) and an amorphous oxide shell, which is silicon oxide with different oxidation states SiOx (0 ≤ x ≤ 2). The samples had an iron content of 0.8–1.8 at %. Colloidal solutions of the nanoparticles stabilized by citrate anions were obtained and characterized. According to the analysis of the cytotoxicity of the modified nanosilicon particles using monoclonal K562 human erythroleukemia cells, no toxicity was found for cells in culture at particle concentrations of up to 5 µg/mL.Conclusions. Since the obtained modified particles are nontoxic, they can be used in in vivo theranostic applications.","PeriodicalId":12215,"journal":{"name":"Fine Chemical Technologies","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81129000","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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