Pub Date : 2024-05-08DOI: 10.33380/2305-2066-2024-13-2-1751
R. Terekhov, A. Taldaev, E. V. Bocharov, D. I. Pankov, A. D. Savina, I. A. Selivanova
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.
{"title":"Analysis of cis-isomer-enriched dihydroquercetin sample by 1D and 2D NMR spectroscopy","authors":"R. Terekhov, A. Taldaev, E. V. Bocharov, D. I. Pankov, A. D. Savina, I. A. Selivanova","doi":"10.33380/2305-2066-2024-13-2-1751","DOIUrl":"https://doi.org/10.33380/2305-2066-2024-13-2-1751","url":null,"abstract":"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.","PeriodicalId":11259,"journal":{"name":"Drug development & registration","volume":"12 10","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141001309","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 : 2024-05-08DOI: 10.33380/2305-2066-2024-13-2-1796
A. Y. Sokolova, A. Poluyanov, A. Bardakov, S. Sologova, N. V. Bobkova
Introduction. Search for new plant species containing biologically active substances (hereinafter – BAS) is one of the leading tasks of pharmacognosy as a science. The search for flavonoid glycosides in plant raw materials is especially relevant, since they have anti-inflammatory, antioxidant, immunostimulating, as well as weak fungicidal and bacteriostatic action. Staphylea pinnata L. is an endemic plant of the Caucasus, cultivated not only in Georgia, but also in the Russian Federation in the Northern and Northwestern Caucasus. In foreign literature there are studies of antioxidant, antiproliferative and cytotoxic activity of leaf extracts of several species of S. pinnata L., as well as inhibitory activity of COX-1, COX-2 and LTB4 formation. Meanwhile, no serious Russian-language scientific studies on either the chemical composition or pharmacological action of generative organs of S. pinnata were found in the literature. This work is part of a comprehensive phytochemical study of S. pinnata. The aim of the work is to study the qualitative and quantitative composition of flavonoids in the studied object.Aim. To isolate, identify and quantify flavonoids in flowers and buds of Staphylea pinnata L.Materials and methods. Alcohol-water extracts from dried generative organs of the studied plant were used as analyzed solutions. Solutions were analyzed on a spectrophotometer SF-2000 (LLC "OKB Spectr", Russia) after sample preparation with aluminum chloride and on an HPLC Nexera-i LC-2040 (Shimadzu Corporation, Japan) equipped with a column and sample thermostat, degasser and autosampler using an individually selected elution gradient of the mobile phase (0.1 % orthophosphoric acid/acetonitrile solution). The primary data were processed using LabSolutions Single LC software (Shimadzu Corporation, Japan). Compounds from the flavonoid group were identified by retention times. Detection was carried out using a UV detector with an absorption wavelength of 365 ± 2 nm.Result and discussion. Alcohol-water extracts from flowers and buds of S. pinnata L. were obtained. Quantitative evaluation by spectrophotometry for flavonoid content was carried out. A gradient elution mode for HPLC was selected for simultaneous determination of 7 flavonoid glycosides. These chromatographic conditions allowed the identification and quantification of astragaline, cynaroside, cosmosiin, narcissin and rutin in flowers and buds of Staphylea pinnata L. Flavonoid glycosides: raponticin and kaempferol were not detected.Conclusion. Flavonoid glycosides were isolated from the generative organs of S. pinnata L., a technique for quantitative determination of flavonoid glycosides in alcohol-water extracts was developed, astragalin, cynaroside, cosmosiin, narcissin and rutin were detected and quantified.
{"title":"Isolation, identification and quantification of flavonoids from the flowers of Staphylea pinnata L.","authors":"A. Y. Sokolova, A. Poluyanov, A. Bardakov, S. Sologova, N. V. Bobkova","doi":"10.33380/2305-2066-2024-13-2-1796","DOIUrl":"https://doi.org/10.33380/2305-2066-2024-13-2-1796","url":null,"abstract":"Introduction. Search for new plant species containing biologically active substances (hereinafter – BAS) is one of the leading tasks of pharmacognosy as a science. The search for flavonoid glycosides in plant raw materials is especially relevant, since they have anti-inflammatory, antioxidant, immunostimulating, as well as weak fungicidal and bacteriostatic action. Staphylea pinnata L. is an endemic plant of the Caucasus, cultivated not only in Georgia, but also in the Russian Federation in the Northern and Northwestern Caucasus. In foreign literature there are studies of antioxidant, antiproliferative and cytotoxic activity of leaf extracts of several species of S. pinnata L., as well as inhibitory activity of COX-1, COX-2 and LTB4 formation. Meanwhile, no serious Russian-language scientific studies on either the chemical composition or pharmacological action of generative organs of S. pinnata were found in the literature. This work is part of a comprehensive phytochemical study of S. pinnata. The aim of the work is to study the qualitative and quantitative composition of flavonoids in the studied object.Aim. To isolate, identify and quantify flavonoids in flowers and buds of Staphylea pinnata L.Materials and methods. Alcohol-water extracts from dried generative organs of the studied plant were used as analyzed solutions. Solutions were analyzed on a spectrophotometer SF-2000 (LLC \"OKB Spectr\", Russia) after sample preparation with aluminum chloride and on an HPLC Nexera-i LC-2040 (Shimadzu Corporation, Japan) equipped with a column and sample thermostat, degasser and autosampler using an individually selected elution gradient of the mobile phase (0.1 % orthophosphoric acid/acetonitrile solution). The primary data were processed using LabSolutions Single LC software (Shimadzu Corporation, Japan). Compounds from the flavonoid group were identified by retention times. Detection was carried out using a UV detector with an absorption wavelength of 365 ± 2 nm.Result and discussion. Alcohol-water extracts from flowers and buds of S. pinnata L. were obtained. Quantitative evaluation by spectrophotometry for flavonoid content was carried out. A gradient elution mode for HPLC was selected for simultaneous determination of 7 flavonoid glycosides. These chromatographic conditions allowed the identification and quantification of astragaline, cynaroside, cosmosiin, narcissin and rutin in flowers and buds of Staphylea pinnata L. Flavonoid glycosides: raponticin and kaempferol were not detected.Conclusion. Flavonoid glycosides were isolated from the generative organs of S. pinnata L., a technique for quantitative determination of flavonoid glycosides in alcohol-water extracts was developed, astragalin, cynaroside, cosmosiin, narcissin and rutin were detected and quantified.","PeriodicalId":11259,"journal":{"name":"Drug development & registration","volume":" 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140998531","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 : 2024-05-08DOI: 10.33380/2305-2066-2024-13-2-1617
S. Bourakba, A. Marakhova, Ya. M. Stanishevskiy, I. A. Vasilenko, V. Zhilkina
Introduction. Foeniculum vulgare Mill., commonly known as fennel or drugstore dill, is very common as one of the world's oldest spice plants, which has significant economic importance and wide application in the pharmaceutical industry. In medical practice, the fruits of this plant are mainly used, which are included in the State Pharmacopoeia of the XIV edition and are a source of essential oil. They are used as an antispasmodic and carminative agent in the form of an infusion. Fennel grass harvested before flowering is used for food, as well as fruits, stems and inflorescences of the plant as a seasoning.Text. The purpose of this study is to summarize scientific papers on the morphological characteristics, phytochemical compounds, therapeutic properties and basic mechanisms of pharmacological activity of F. vulgare. A systematic literature search was conducted using relevant keywords such as "fennel", "Foeniculum vulgare Mill.", "therapeutic" and "pharmacology" in well-known databases, including ScienceDirect, Scopus, EBSCO and Medline. The search covered articles published before April 25, 2023 in available journals. The results of the study showed that fennel has a wide range of pharmacological properties, including antioxidant, anti-cancer, anti-inflammatory, antifungal, antibacterial and estrogenic effects. These effects can be explained by the presence of aromatic compounds, in particular anethole, estragole and fenchone, which are abundantly present in fennel.Conclusion. The diverse pharmacological properties and rich chemical composition of fennel make it a promising raw material for the development of new medicines. Further study of the therapeutic potential of fennel through in vivo and in vitro studies will contribute to the establishment of mechanisms of pharmacological action of biologically active substances (BAS) of fennel.
{"title":"Exploring the potential of fennel (Foeniculum vulgare Mill.) as a valuable medicinal plant: a comprehensive (review)","authors":"S. Bourakba, A. Marakhova, Ya. M. Stanishevskiy, I. A. Vasilenko, V. Zhilkina","doi":"10.33380/2305-2066-2024-13-2-1617","DOIUrl":"https://doi.org/10.33380/2305-2066-2024-13-2-1617","url":null,"abstract":"Introduction. Foeniculum vulgare Mill., commonly known as fennel or drugstore dill, is very common as one of the world's oldest spice plants, which has significant economic importance and wide application in the pharmaceutical industry. In medical practice, the fruits of this plant are mainly used, which are included in the State Pharmacopoeia of the XIV edition and are a source of essential oil. They are used as an antispasmodic and carminative agent in the form of an infusion. Fennel grass harvested before flowering is used for food, as well as fruits, stems and inflorescences of the plant as a seasoning.Text. The purpose of this study is to summarize scientific papers on the morphological characteristics, phytochemical compounds, therapeutic properties and basic mechanisms of pharmacological activity of F. vulgare. A systematic literature search was conducted using relevant keywords such as \"fennel\", \"Foeniculum vulgare Mill.\", \"therapeutic\" and \"pharmacology\" in well-known databases, including ScienceDirect, Scopus, EBSCO and Medline. The search covered articles published before April 25, 2023 in available journals. The results of the study showed that fennel has a wide range of pharmacological properties, including antioxidant, anti-cancer, anti-inflammatory, antifungal, antibacterial and estrogenic effects. These effects can be explained by the presence of aromatic compounds, in particular anethole, estragole and fenchone, which are abundantly present in fennel.Conclusion. The diverse pharmacological properties and rich chemical composition of fennel make it a promising raw material for the development of new medicines. Further study of the therapeutic potential of fennel through in vivo and in vitro studies will contribute to the establishment of mechanisms of pharmacological action of biologically active substances (BAS) of fennel.","PeriodicalId":11259,"journal":{"name":"Drug development & registration","volume":"5 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141001528","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 : 2024-05-03DOI: 10.33380/2305-2066-2024-13-2-1682
E. Avdeeva, M. Skorokhodova, V. V. Sheikin, T. V. Kadyrova, M. V. Belousov
Introduction. Medicines of natural origin have advantages over synthetic drugs and occupy a significant share in the assortment of pharmacies. Filipendula ulmaria (L.) Maxim., characterized by a chemical composition rich in phenolic compounds and various types of biological activity has the potential in terms of the development of new herbal medicines. In an in vivo experiment, F. ulmaria extract showed immunomodulatory, anti-inflammatory, osteogenic activity and effectively stimulated myelopoiesis in conditions of microbial damage of bone tissue.Aim. Development of an optimal technology for obtaining dry extract of F. ulmaria by remaceration and investigation of its аcute toxicity.Materials and methods. The technology of extract was developed in a laboratory reactor equipped with a water heater jacket and a stirrer. The content of flavonoids was determined by differential spectrometry. Flowability was determined on a flowability tester for powders and granules, bulk density was determined on a bulk density tester for powders and granules, moisture content was developed on a moisture content analyzer. Acute toxicity was determined on Wistar rats.Results and discussion. The dry extract of F. ulmaria, obtained using 70 % ethanol by remaceration, is effective in conditions of microbial damage of bone tissue and bone marrow, which determined the choice of extragent and the type of extract. The optimal parameters for obtaining the extract were determined (three-fold maceration at a water jacket temperature of 80 °C, the ratio of raw materials to extragent 1 : 14, time 30 min), its main technological characteristics were determined and a wet granulation method for improving them was proposed. In the study of acute toxicity of the extract, according to the results of toxicometry for 14 days after administration and necropsy data, the studied extract was classified as low-toxic substance.Conclusion. The dry extract of F. ulmaria, obtained by remaceration using 70 % ethanol, has specific activity in conditions of microbial damage to bone tissue. Optimal extraction parameters allow to increase the yield of target components, it is advisable to improve the technological parameters of the extract by wet granulation. The assessment of the safety profile of the dry extract from F. ulmaria, along with its high biological activity, provides a perspective for its further study and implementation into medical practice.
{"title":"Technology of dry extract from the Filipendula ulmaria (L.) Maxim. and preliminary assessment of its safety profile","authors":"E. Avdeeva, M. Skorokhodova, V. V. Sheikin, T. V. Kadyrova, M. V. Belousov","doi":"10.33380/2305-2066-2024-13-2-1682","DOIUrl":"https://doi.org/10.33380/2305-2066-2024-13-2-1682","url":null,"abstract":"Introduction. Medicines of natural origin have advantages over synthetic drugs and occupy a significant share in the assortment of pharmacies. Filipendula ulmaria (L.) Maxim., characterized by a chemical composition rich in phenolic compounds and various types of biological activity has the potential in terms of the development of new herbal medicines. In an in vivo experiment, F. ulmaria extract showed immunomodulatory, anti-inflammatory, osteogenic activity and effectively stimulated myelopoiesis in conditions of microbial damage of bone tissue.Aim. Development of an optimal technology for obtaining dry extract of F. ulmaria by remaceration and investigation of its аcute toxicity.Materials and methods. The technology of extract was developed in a laboratory reactor equipped with a water heater jacket and a stirrer. The content of flavonoids was determined by differential spectrometry. Flowability was determined on a flowability tester for powders and granules, bulk density was determined on a bulk density tester for powders and granules, moisture content was developed on a moisture content analyzer. Acute toxicity was determined on Wistar rats.Results and discussion. The dry extract of F. ulmaria, obtained using 70 % ethanol by remaceration, is effective in conditions of microbial damage of bone tissue and bone marrow, which determined the choice of extragent and the type of extract. The optimal parameters for obtaining the extract were determined (three-fold maceration at a water jacket temperature of 80 °C, the ratio of raw materials to extragent 1 : 14, time 30 min), its main technological characteristics were determined and a wet granulation method for improving them was proposed. In the study of acute toxicity of the extract, according to the results of toxicometry for 14 days after administration and necropsy data, the studied extract was classified as low-toxic substance.Conclusion. The dry extract of F. ulmaria, obtained by remaceration using 70 % ethanol, has specific activity in conditions of microbial damage to bone tissue. Optimal extraction parameters allow to increase the yield of target components, it is advisable to improve the technological parameters of the extract by wet granulation. The assessment of the safety profile of the dry extract from F. ulmaria, along with its high biological activity, provides a perspective for its further study and implementation into medical practice.","PeriodicalId":11259,"journal":{"name":"Drug development & registration","volume":"80 11","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141016303","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 : 2024-04-22DOI: 10.33380/2305-2066-2024-13-2-1638
N. Popov, A. A. Antipina, S. V. Savintsev, V. Balabanyan
Introduction. Polyprenols are known as a class of natural long-chain isoprenoid alcohols, which are natural bioregulators that directly participate in the synthesis of cell membrane glycoproteins. Their hepatoprotective activity is proven, as well other types of their pharmacological effects are known, which is the reason of significant interest in these substances as a promising medicinal product. It is non-trivial task to determine the sum of polyprenols in extracts as include design and implementation of accurate reproducible analytical methods, which will subsequently be used in standardization.Aim. Development and validation of the chromatographic-mass spectrometric technique for polyprenols identification and their quantitative assessment.Materials and methods. Chromatographic separation of polyprenols was performed by using an HPLC Agilent 1260 Infinity II (Agilent Technologies, США); with the mixture of methanol, n-hexane, propanol-2, and aqueous ammonium acetate solution as eluent in gradient mode. An AB Sciex QTrap® 3200MD (AB Sciex Pte. Ltd., Singapore) triple quadrupole mass spectrometer was used as a detector, with the registration of polyprenols adducts.Results and discussion. The conditions for chromatographic separation and detection of polyprenols were identified. The developed technique was validated for the following characteristics: specificity, limit of detection, limit of quantification, linearity, accuracy, precision, range of application, and stability.Conclusion. It was determined the content of polyprenols in the substance recieved from Ginkgo biloba L. and Picea abies L. The developed technique can be used in the future to assess the content of polyprenols in drug products or pharmaceutical substances.
{"title":"Development and validation of the chromatography-mass spectrometry technique for polyprenols quantitative assessment","authors":"N. Popov, A. A. Antipina, S. V. Savintsev, V. Balabanyan","doi":"10.33380/2305-2066-2024-13-2-1638","DOIUrl":"https://doi.org/10.33380/2305-2066-2024-13-2-1638","url":null,"abstract":"Introduction. Polyprenols are known as a class of natural long-chain isoprenoid alcohols, which are natural bioregulators that directly participate in the synthesis of cell membrane glycoproteins. Their hepatoprotective activity is proven, as well other types of their pharmacological effects are known, which is the reason of significant interest in these substances as a promising medicinal product. It is non-trivial task to determine the sum of polyprenols in extracts as include design and implementation of accurate reproducible analytical methods, which will subsequently be used in standardization.Aim. Development and validation of the chromatographic-mass spectrometric technique for polyprenols identification and their quantitative assessment.Materials and methods. Chromatographic separation of polyprenols was performed by using an HPLC Agilent 1260 Infinity II (Agilent Technologies, США); with the mixture of methanol, n-hexane, propanol-2, and aqueous ammonium acetate solution as eluent in gradient mode. An AB Sciex QTrap® 3200MD (AB Sciex Pte. Ltd., Singapore) triple quadrupole mass spectrometer was used as a detector, with the registration of polyprenols adducts.Results and discussion. The conditions for chromatographic separation and detection of polyprenols were identified. The developed technique was validated for the following characteristics: specificity, limit of detection, limit of quantification, linearity, accuracy, precision, range of application, and stability.Conclusion. It was determined the content of polyprenols in the substance recieved from Ginkgo biloba L. and Picea abies L. The developed technique can be used in the future to assess the content of polyprenols in drug products or pharmaceutical substances.","PeriodicalId":11259,"journal":{"name":"Drug development & registration","volume":"47 17","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140672793","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 : 2024-04-19DOI: 10.33380/2305-2066-2024-13-2-1715
O. V. Trineeva, O. V. Pugacheva
{"title":"Profile of biologically active substances of Aronia mitschurinii leaves growing in the conditions of the Central Black Earth region","authors":"O. V. Trineeva, O. V. Pugacheva","doi":"10.33380/2305-2066-2024-13-2-1715","DOIUrl":"https://doi.org/10.33380/2305-2066-2024-13-2-1715","url":null,"abstract":"","PeriodicalId":11259,"journal":{"name":"Drug development & registration","volume":" 11","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140684483","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 : 2024-04-15DOI: 10.33380/2305-2066-2024-13-2-1783
M. A. Tokareva, E. Melnikov, M. V. Belova, E. Fisher, T. Rodina, I. Shohin
Introduction. The number of peptide drugs being developed and registered has increased in recent years. Therefore, modern analytical approaches and methods are required to determine these substances in biological matrices during pharmacokinetic studies. Peptides are structurally intermediate between small molecules and biopolymers, making it difficult to develop methods for determining them using High Performance Liquid Chromatography with Tandem Mass Spectrometry (HPLC-MS/MS). Peptide derivatization can help achieve optimal chromatographic separation and increase method sensitivity.Aim. To develop and validate a method for the determination of the tetradecapeptide (TDP) threonyl-glutamyl-lysyl-lysyl-arginyl-arginyl-glutamayl-threonyl-valyl-glutamyl-arginyl-glutamyl-lysyl-glutamate in human plasma by HPLC-MS/MS.Materials and methods. The determination of TDP in human plasma was performed by HPLC-MS/MS. Sample preparation included a combination of blood plasma protein precipitation with propionic acid solution in methanol, liquid-liquid extraction with chloroform, and peptide derivatization with propionic anhydride. Internal standard (IS) was threonyl-glutamyl-lysyl-lysyl-arginyl-arginyl-glutamayl-threonyl-leucyl-glutamyl-arginyl-glutamyl-lysyl-glutamate. Chromatographic separation was performed in gradient mode, eluent A was 0.1 % formic acid solution in water, eluent B was 0.1 % formic acid in acetonitrile. Column: Waters XBridge C18, 4.6 × 50 mm, 5 µm. Ionization source was electrospray in positive mode. Multiple reaction monitoring (MRM) transitions for 4-substituted TDP propionate were: 681.30 → 73.95 m/z, 681.30 → 84.00 m/z, 681.30 → 101.90 m/z, 681.30 → 140.10 m/z, and for 4-substituted IS propionate: 686.00 → 74.10 m/z, 686.00 → 84.05 m/z, 686.00 → 102.00 m/z, 686.00 → 140.00 m/z.Results and discussion. Validation of the developed method was carried out in accordance with the requirements of Eurasian Economic Union and the following parameters were determined: selectivity, matrix effect, calibration curve, accuracy and precision, recovery, lower limit of quantification, sample carryover, stability.Conclusion. The method for the determination of TDP in human blood plasma by HPLC-MS/MS was developed and validated. The analytical range was 5.00–1000.00 ng/mL, allowing the method to be used to study TDP pharmacokinetics.
{"title":"HPLC-MS/MS method development and validation for the determination of tetradecapeptide in human plasma","authors":"M. A. Tokareva, E. Melnikov, M. V. Belova, E. Fisher, T. Rodina, I. Shohin","doi":"10.33380/2305-2066-2024-13-2-1783","DOIUrl":"https://doi.org/10.33380/2305-2066-2024-13-2-1783","url":null,"abstract":"Introduction. The number of peptide drugs being developed and registered has increased in recent years. Therefore, modern analytical approaches and methods are required to determine these substances in biological matrices during pharmacokinetic studies. Peptides are structurally intermediate between small molecules and biopolymers, making it difficult to develop methods for determining them using High Performance Liquid Chromatography with Tandem Mass Spectrometry (HPLC-MS/MS). Peptide derivatization can help achieve optimal chromatographic separation and increase method sensitivity.Aim. To develop and validate a method for the determination of the tetradecapeptide (TDP) threonyl-glutamyl-lysyl-lysyl-arginyl-arginyl-glutamayl-threonyl-valyl-glutamyl-arginyl-glutamyl-lysyl-glutamate in human plasma by HPLC-MS/MS.Materials and methods. The determination of TDP in human plasma was performed by HPLC-MS/MS. Sample preparation included a combination of blood plasma protein precipitation with propionic acid solution in methanol, liquid-liquid extraction with chloroform, and peptide derivatization with propionic anhydride. Internal standard (IS) was threonyl-glutamyl-lysyl-lysyl-arginyl-arginyl-glutamayl-threonyl-leucyl-glutamyl-arginyl-glutamyl-lysyl-glutamate. Chromatographic separation was performed in gradient mode, eluent A was 0.1 % formic acid solution in water, eluent B was 0.1 % formic acid in acetonitrile. Column: Waters XBridge C18, 4.6 × 50 mm, 5 µm. Ionization source was electrospray in positive mode. Multiple reaction monitoring (MRM) transitions for 4-substituted TDP propionate were: 681.30 → 73.95 m/z, 681.30 → 84.00 m/z, 681.30 → 101.90 m/z, 681.30 → 140.10 m/z, and for 4-substituted IS propionate: 686.00 → 74.10 m/z, 686.00 → 84.05 m/z, 686.00 → 102.00 m/z, 686.00 → 140.00 m/z.Results and discussion. Validation of the developed method was carried out in accordance with the requirements of Eurasian Economic Union and the following parameters were determined: selectivity, matrix effect, calibration curve, accuracy and precision, recovery, lower limit of quantification, sample carryover, stability.Conclusion. The method for the determination of TDP in human blood plasma by HPLC-MS/MS was developed and validated. The analytical range was 5.00–1000.00 ng/mL, allowing the method to be used to study TDP pharmacokinetics.","PeriodicalId":11259,"journal":{"name":"Drug development & registration","volume":"34 15","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140702255","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 : 2024-04-01DOI: 10.33380/2305-2066-2024-13-2-1611
A. S. Chistyakova, A. Dunilin, O. V. Trineeva, A. S. Bolgov, A. Gudkova, A. I. Slivkin
Introduction. Currently, there is an increasing trend towards assessing the full spectrum of chemical compounds in plant materials. One of the first steps in the formation of secondary metabolites of complex structure, participating in biochemical processes in the plant body, which have their own effect on the human and animal body, are organic acids (OA) and amino acids (AA). However, due attention is not always given to these compounds when studying the component composition of plant raw materials.Aim. The purpose of the work was to study the profile and quantitative assessment of the content of organic acids and amino acids of Aesculus hippocastanum L. flowers.Materials and methods. The object of the study were horse chestnut flowers, collected during flowering in the Voronezh region in 2021 and dried by air-shade method. Quantitative assessment of the amount of TC in terms of malic acid and ascorbic acid was carried out titrimetrically according to the methods presented in the State Pharmacopoeia of the Russian Federation, XIV edition. The composition of the OA and AA profiles of horse chestnut flowers was determined using the capillary electrophoresis method ("Kapel®-105/105M", Group of Companies "Lumex", Russia).Results and discussion. The method of capillary electrophoresis revealed the presence of 12 compounds of the OA group in a total amount of 4.7 % (in terms of absolutely dry raw materials). The rationality of recalculating the content of the sum of OA to the major component – citric acid, which amounted to 1.65 ± 0.04 %, was shown. The content of ascorbic acid was low (0.033 ± 0.007) %. Using the TLC method, the presence of 5 zones of compounds belonging to the AA class was established in the aqueous extract of horse chestnut flowers. The quantitative content of the sum of free AA in terms of glutamine, determined by the spectrophotometric method, was 2.25 ± 0.07 % (n = 5). The presence of 17 AA was determined by capillary electrophoresis, which amounted to 7.13 %. Glutamic acid is present in the greatest quantity in horse chestnut flowers (1.19 %). Essential amino acids amounted to 2.27 %; leucine was present in the predominant amount (0.58 %).Conclusion. A study was carried out of the qualitative composition and quantitative content of organic acids and amino acids of horse chestnut flowers. The composition of the profile and the quantitative content of organic acids and amino acids have been established. It was revealed that citric acid prevails in the total organic acids; glutamic, aspartic acid, arginine and proline are found in greater quantities in the total amino acids.
导言。目前，人们越来越倾向于对植物材料中的所有化合物进行评估。有机酸（OA）和氨基酸（AA）是形成结构复杂的次级代谢产物的第一步，它们参与植物体内的生化过程，并对人体和动物产生影响。然而，在研究植物原材料的成分组成时，人们并不总是对这些化合物给予应有的重视。这项工作的目的是研究马胸花（Aesculus hippocastanum L.）有机酸和氨基酸含量的概况和定量评估。研究对象是 2021 年在沃罗涅日地区花期采集的七叶树花，采用遮光法干燥。根据《俄罗斯联邦国家药典》（第 XIV 版）规定的方法，用滴定法对苹果酸和抗坏血酸的 TC 含量进行了定量评估。使用毛细管电泳法（"Kapel®-105/105M"，集团公司 "Lumex"，俄罗斯）测定了七叶树花的 OA 和 AA 组成。毛细管电泳法检测出 12 种 OA 类化合物，总含量为 4.7%（按绝对干燥原料计）。将 OA 的总和含量重新计算为主要成分柠檬酸（1.65 ± 0.04 %）的合理性得到了证明。抗坏血酸的含量较低（0.033 ± 0.007）%。通过 TLC 法，确定了在七叶树花的水提取物中存在 5 个属于 AA 类化合物的区域。用分光光度法测定的谷氨酰胺游离 AA 总量的定量含量为 2.25 ± 0.07 %（n = 5）。用毛细管电泳法测定的 17 AA 含量为 7.13%。谷氨酸在七叶树花中的含量最高（1.19 %）。必需氨基酸含量为 2.27%；亮氨酸含量最多（0.58%）。对七叶树花的有机酸和氨基酸的定性组成和定量含量进行了研究。确定了有机酸和氨基酸的概况组成和定量含量。研究表明，柠檬酸在有机酸总量中占主导地位；谷氨酸、天门冬氨酸、精氨酸和脯氨酸在氨基酸总量中含量较高。
{"title":"Studying the acids and amino acid composition of horse chestnut flowers","authors":"A. S. Chistyakova, A. Dunilin, O. V. Trineeva, A. S. Bolgov, A. Gudkova, A. I. Slivkin","doi":"10.33380/2305-2066-2024-13-2-1611","DOIUrl":"https://doi.org/10.33380/2305-2066-2024-13-2-1611","url":null,"abstract":"Introduction. Currently, there is an increasing trend towards assessing the full spectrum of chemical compounds in plant materials. One of the first steps in the formation of secondary metabolites of complex structure, participating in biochemical processes in the plant body, which have their own effect on the human and animal body, are organic acids (OA) and amino acids (AA). However, due attention is not always given to these compounds when studying the component composition of plant raw materials.Aim. The purpose of the work was to study the profile and quantitative assessment of the content of organic acids and amino acids of Aesculus hippocastanum L. flowers.Materials and methods. The object of the study were horse chestnut flowers, collected during flowering in the Voronezh region in 2021 and dried by air-shade method. Quantitative assessment of the amount of TC in terms of malic acid and ascorbic acid was carried out titrimetrically according to the methods presented in the State Pharmacopoeia of the Russian Federation, XIV edition. The composition of the OA and AA profiles of horse chestnut flowers was determined using the capillary electrophoresis method (\"Kapel®-105/105M\", Group of Companies \"Lumex\", Russia).Results and discussion. The method of capillary electrophoresis revealed the presence of 12 compounds of the OA group in a total amount of 4.7 % (in terms of absolutely dry raw materials). The rationality of recalculating the content of the sum of OA to the major component – citric acid, which amounted to 1.65 ± 0.04 %, was shown. The content of ascorbic acid was low (0.033 ± 0.007) %. Using the TLC method, the presence of 5 zones of compounds belonging to the AA class was established in the aqueous extract of horse chestnut flowers. The quantitative content of the sum of free AA in terms of glutamine, determined by the spectrophotometric method, was 2.25 ± 0.07 % (n = 5). The presence of 17 AA was determined by capillary electrophoresis, which amounted to 7.13 %. Glutamic acid is present in the greatest quantity in horse chestnut flowers (1.19 %). Essential amino acids amounted to 2.27 %; leucine was present in the predominant amount (0.58 %).Conclusion. A study was carried out of the qualitative composition and quantitative content of organic acids and amino acids of horse chestnut flowers. The composition of the profile and the quantitative content of organic acids and amino acids have been established. It was revealed that citric acid prevails in the total organic acids; glutamic, aspartic acid, arginine and proline are found in greater quantities in the total amino acids.","PeriodicalId":11259,"journal":{"name":"Drug development & registration","volume":"353 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140778662","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 : 2024-03-25DOI: 10.33380/2305-2066-2024-13-2-1727
E. Bakhrushina, I. Gravel, O. S. Filippova, V. N. Tychinin, A. A. Popova, O. B. Dobrovolsky
Introduction. The secretome of mesenchymal stem cells (SMSC) is widely used in medicine. It is most often used due to its immune-modulating and regenerative properties in the treatment of autoimmune, immuno-mediated and other diseases due to its anti-inflammatory, neuroprotective and regenerating action. In many studies, exosomes isolated from SMSC are used as a therapeutic agent. In recent years, the interest in the development of products containing SMSC for external use has increased. Similar drugs are planned to be used in the treatment of diabetic wounds, for skin regeneration, the treatment of inflammatory diseases, as well as alopecia. There are multiple studies on increasing collagen secretion and reducing skin photosensitivity in preclinical studies, which confirms the significant potential for the use of SMSC in dermatology and cosmetology. The purpose of this review was to study the potential of using conditioned medium in medicines for external use, approaches to standardization of SMSC as a pharmaceutical substance and methods of increasing percutaneous delivery.Text. SMSC as an active pharmaceutical ingredient is a transparent liquid from yellow to orange in color with a characteristic odor. The pH of the ready-to-use SMSC composition ranges from 7.0 to 7.5, which allows it to be used in topical and external applications without the addition of stabilizers or pH correctors. Problems of delivery of SMSC through the epidermis are most often solved by placing the secretome in hydrogels, using exosomes or technology using microneedles. Since 2022, after legislative changes, measures have been taken to register and introduce into clinical practice domestic drugs based on cellular products. However, as the analysis showed, it will take some time before the appearance of original medicines based on SMSC, and today in the Russian Federation only products related to cosmetics and veterinary drugs, as well as zoocosmetics, are produced so far.Conclusion. SMSC may also prove to be a safer and more effective substance for the potential treatment of a wide range of acute and chronic diseases. But despite the large number of positive results of using SMSC for wound healing in animals, as well as clinical studies on skin regeneration, there are no studies of its safety and effectiveness, as well as standardization of the production process.
{"title":"Modern Aspects of External Application and Prospects of Using the Secretome of Mesenchymal Stem Cells (Review)","authors":"E. Bakhrushina, I. Gravel, O. S. Filippova, V. N. Tychinin, A. A. Popova, O. B. Dobrovolsky","doi":"10.33380/2305-2066-2024-13-2-1727","DOIUrl":"https://doi.org/10.33380/2305-2066-2024-13-2-1727","url":null,"abstract":"Introduction. The secretome of mesenchymal stem cells (SMSC) is widely used in medicine. It is most often used due to its immune-modulating and regenerative properties in the treatment of autoimmune, immuno-mediated and other diseases due to its anti-inflammatory, neuroprotective and regenerating action. In many studies, exosomes isolated from SMSC are used as a therapeutic agent. In recent years, the interest in the development of products containing SMSC for external use has increased. Similar drugs are planned to be used in the treatment of diabetic wounds, for skin regeneration, the treatment of inflammatory diseases, as well as alopecia. There are multiple studies on increasing collagen secretion and reducing skin photosensitivity in preclinical studies, which confirms the significant potential for the use of SMSC in dermatology and cosmetology. The purpose of this review was to study the potential of using conditioned medium in medicines for external use, approaches to standardization of SMSC as a pharmaceutical substance and methods of increasing percutaneous delivery.Text. SMSC as an active pharmaceutical ingredient is a transparent liquid from yellow to orange in color with a characteristic odor. The pH of the ready-to-use SMSC composition ranges from 7.0 to 7.5, which allows it to be used in topical and external applications without the addition of stabilizers or pH correctors. Problems of delivery of SMSC through the epidermis are most often solved by placing the secretome in hydrogels, using exosomes or technology using microneedles. Since 2022, after legislative changes, measures have been taken to register and introduce into clinical practice domestic drugs based on cellular products. However, as the analysis showed, it will take some time before the appearance of original medicines based on SMSC, and today in the Russian Federation only products related to cosmetics and veterinary drugs, as well as zoocosmetics, are produced so far.Conclusion. SMSC may also prove to be a safer and more effective substance for the potential treatment of a wide range of acute and chronic diseases. But despite the large number of positive results of using SMSC for wound healing in animals, as well as clinical studies on skin regeneration, there are no studies of its safety and effectiveness, as well as standardization of the production process.","PeriodicalId":11259,"journal":{"name":"Drug development & registration","volume":"12 13","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140381724","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 : 2024-03-25DOI: 10.33380/2305-2066-2024-13-2-1714
F. D. Evsikov, A. Gudkova, A. I. Slivkin
Introduction. Virgin grape (maiden grape, Virginia grape, virgin grape, Parthenocissus quinquefolia (L.) Planch.) is currently used primarily for decorative purposes, but is also beginning to attract scientists from different countries as a promising source of important compounds such as anthocyanins, flavonoids, organic acids, macro- and microelements. To introduce a plant into medical practice, one of the first steps is the need to develop regulatory documentation that allows for quality assessment and standardization of plant materials, and the first stage of this process is the assessment of the characteristics of the authenticity of the object.Aim. Establish the morphological characteristics of the leaf and petiole of the virgin grape (Parthenocissus quinquefolia (L.) Planch.).Materials and methods. The object of the study was developed leaves of the virgin grape, harvested in the Voronezh region in the summer of 2022 and dried in the shade. To carry out the experiment, a JSM-6510LV electron microscope was used; the analysis was carried out after sputtering a thin film of gold onto the sample.Results and discussion. For the first time, the main structural features of various morphological features of the leaf blade of the virgin grape were identified, differences in the morphological picture of the upper and lower sides of the leaf were established, and structural features of the surface of the petiole were identified. Using micro-X-ray analysis, it was revealed that calcium and potassium are present in large quantities in the leaves and petiole of virgin grapes. The least amount of the studied elements in the study object contains magnesium. It has been shown that the leaves contain a small amount of chlorine, and the cellular contents of the petiole include a small amount of sodium.Conclusion. For the first time, a study of the morphology of the surface of the leaf blade and petiole of the virgin grape was carried out using scanning electron microscopy. The main diagnostic features of plant raw materials have been identified and visualized. Micro-X-ray structural analysis made it possible to establish the composition of the elements of the leaf blade and petiole of virgin grapes and revealed the ability to concentrate calcium, potassium, and phosphorus.
导言。处女葡萄（少女葡萄、弗吉尼亚葡萄、处女葡萄、Parthenocissus quinquefolia (L.) Planch.）目前主要用于装饰目的，但作为花青素、类黄酮、有机酸、宏量和微量元素等重要化合物的一个有前途的来源，也开始吸引各国科学家的关注。要将一种植物引入医疗实践，首要步骤之一是需要制定监管文件，以便对植物材料进行质量评估和标准化，而这一过程的第一阶段就是评估对象的真实性特征。确定原生葡萄（Parthenocissus quinquefolia (L.) Planch.）叶片和叶柄的形态特征。研究对象是 2022 年夏季在沃罗涅日地区收获并在阴凉处晾干的原生葡萄叶片。实验使用了 JSM-6510LV 电子显微镜，在样品上溅射一层金薄膜后进行分析。首次确定了原生葡萄叶片各种形态特征的主要结构特征，确定了叶片上下两面形态图谱的差异，并确定了叶柄表面的结构特征。通过显微 X 射线分析发现，钙和钾大量存在于原生葡萄的叶片和叶柄中。研究对象中的镁元素含量最少。研究表明，叶片中含有少量的氯，叶柄的细胞成分中含有少量的钠。首次使用扫描电子显微镜对原生葡萄叶片和叶柄表面的形态进行了研究。对植物原材料的主要诊断特征进行了鉴定和可视化。显微 X 射线结构分析使确定原生葡萄叶片和叶柄的元素组成成为可能，并揭示了浓缩钙、钾和磷的能力。
{"title":"Raster Electron Microscopy in the Analysis of Plant Raw Materials of Virgin Grapes (Parthenocissus quinquefolia (L.) Planch.)","authors":"F. D. Evsikov, A. Gudkova, A. I. Slivkin","doi":"10.33380/2305-2066-2024-13-2-1714","DOIUrl":"https://doi.org/10.33380/2305-2066-2024-13-2-1714","url":null,"abstract":"Introduction. Virgin grape (maiden grape, Virginia grape, virgin grape, Parthenocissus quinquefolia (L.) Planch.) is currently used primarily for decorative purposes, but is also beginning to attract scientists from different countries as a promising source of important compounds such as anthocyanins, flavonoids, organic acids, macro- and microelements. To introduce a plant into medical practice, one of the first steps is the need to develop regulatory documentation that allows for quality assessment and standardization of plant materials, and the first stage of this process is the assessment of the characteristics of the authenticity of the object.Aim. Establish the morphological characteristics of the leaf and petiole of the virgin grape (Parthenocissus quinquefolia (L.) Planch.).Materials and methods. The object of the study was developed leaves of the virgin grape, harvested in the Voronezh region in the summer of 2022 and dried in the shade. To carry out the experiment, a JSM-6510LV electron microscope was used; the analysis was carried out after sputtering a thin film of gold onto the sample.Results and discussion. For the first time, the main structural features of various morphological features of the leaf blade of the virgin grape were identified, differences in the morphological picture of the upper and lower sides of the leaf were established, and structural features of the surface of the petiole were identified. Using micro-X-ray analysis, it was revealed that calcium and potassium are present in large quantities in the leaves and petiole of virgin grapes. The least amount of the studied elements in the study object contains magnesium. It has been shown that the leaves contain a small amount of chlorine, and the cellular contents of the petiole include a small amount of sodium.Conclusion. For the first time, a study of the morphology of the surface of the leaf blade and petiole of the virgin grape was carried out using scanning electron microscopy. The main diagnostic features of plant raw materials have been identified and visualized. Micro-X-ray structural analysis made it possible to establish the composition of the elements of the leaf blade and petiole of virgin grapes and revealed the ability to concentrate calcium, potassium, and phosphorus.","PeriodicalId":11259,"journal":{"name":"Drug development & registration","volume":"11 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140381349","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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