Development of Genistein Synthesis for Use as a Certified Reference Material

Introduction. The use of certified reference materials (CRMs) ensures metrological traceability and comparability of analysis results performed in different laboratories, by different analysts, at different times. Genistein is a promising substance with a wide spectrum of pharmacological action. genistein is widely used in dietary supplements. Development of regulatory documents for CRM of genistein will ensure the quality of drugs and dietary supplements.Aim. Aim of our study is to improve of the ways of synthesis and determination of spectrum characteristics of genistein for the certification of CRM.Materials and methods. We used synthetic genistein, (Ph.D. V. Yu. Kovtun SPC "Pharmzashchita") (sample № 1) and genistein synthesized and studied at the departments of pharmaceutical chemistry and chemical technology of medicinal substances SPCPU (sample № 2). Infrared spectra of genistein samples were collected on an FSM 1201 infrared Fourier spectrometer (OOO Infraspek, Russia) via KBr pellets technique. All the spectra were collected in the 4000–500 cm−1 range. The NMR (1H and 13C) measurements were performed with a BrukerAvance III NMR spectrometer (400 and 100 MHz) (Bruker, Germany) in DMSO-d6 solvent. Raman spectra were recorded by an ORTES-785TRS-2700 analytical Raman scattering system at a laser power of 100 mW (OPTEC JSC, Russia). Laser interaction time was 5, 10, 20 and 60 seconds. The results were processed using the software "BWSpec 4.10_4", USA. GC-MS was performed on an Agilent Technologies 7890A gas chromatograph (Agilent Technologies, USA) with a 7693 autoinjector and a Hewlett Packard 5975C mass selective detector.Results and discussion. The synthesis was carried out according to the developer's method. The stage "removal of the alkyl protection" has been improved. The spectra of the synthesis intermediate of genistein (biochanin A) correspond to the literature data. Samples of genistein were investigated by methods: MC and NMR 13С, 1Н. The structure of the investigated substance was confirmed; Raman and IR spectroscopy showed that the spectra of the samples do not differ from each other and there are no additional signals.Conclusion. The spectrum characteristics of samples of genistein were obtained by NMR, IR and Raman spectroscopy, which will be used in the regulatory documentation for CRM of genistein. All of this will make it possible to control the quality of medicines based on it and to identify substandard dietary supplements.

Synthesis of non-phosphorylated epoxidised corn oil as a novel green flame retardant thermoset resin

This study aimed to produce a new potential flame retardant thermoset resin from epoxidised corn oil through a one-pot method using liquid inorganic catalysed with hydrogen peroxide. Using a gas chromatography–mass selective detector, attenuated total reflectance-fourier transform infrared spectroscopy, proton nuclear magnetic resonance imaging, optical microscopy, and scanning emission microscopy, we synthesised a bio-based resin based on newly designed parameters. The flame retardant capacity was fully established using thermogravimetric analysis and a micro calorimeter. The produced epoxidised corn oil had a relative percentage conversion of oxirane of approximately 91.70%, wherein the amount of double bonds converted into epoxides was calculated. A significant reduction from 17 to 40% in peak heat rate release (pHRR) and 26–30% in total heat release was observed, confirming its flame retardant property. Thus, the potential of epoxidised corn oil was demonstrated.

Toxicological Stability of Ocimum basilicum Essential Oil and Its Major Components in the Control of Sitophilus zeamais

Essential oils (EOs) are widely recognized as efficient and safe alternatives for controlling pest insects in foods. However, there is a lack of studies evaluating the toxicological stability of botanical insecticides in stored grains in order to establish criteria of use and ensure your efficiency. The objective of this work was to evaluate the toxicological stability of basil essential oil (O. basilicum) and its linalool and estragole components for Sitophilus zeamais (Motschulsky) adults in corn grains by fumigation. The identification of the chemical compounds of the essential oil was performed with a gas chromatograph coupled to a mass selective detector. Mortality of insects was assessed after 24 h exposure. After storage for six (EO) and two months (linalool and estragole) under different conditions of temperature (5, 20, and 35 °C) and light (with and without exposure to light), its toxicological stability was evaluated. Studies revealed that the essential oil of O. basilicum and its main components exhibited insecticidal potential against adults of S. zeamais. For greater toxicological stability, suitable storage conditions for them include absence of light and temperatures equal to or less than 20 °C.

Development and validation of the method for the quantitative determination of methotrexate in a transport medium by HPLC-MS/MS

Relevance. BCRP is an efflux transporter protein that plays an important role in the pharmacokinetics of a wide range of drugs. The BCRP activity in vitro experiments is assessed by the transport of transporter protein substrates (methotrexate, etc.) across the bilipid membrane of cells overexpressingBCRP, for example, Caco-2 cells. The aim is to develop and validate a method for the quantitative determination of the BCRP substrate, methotrexate, in the transport medium of Caco-2 cells by HPLC-MS/MS. Methods. The work was performed on an Ultimate 3000 HPLC chromatograph (ThermoFisher, USA) with a TSQ Fortis tandem mass-selective detector (ThermoFisher, USA). The conditions of chromatographic analysis were as follows: column UCT Selectra C18 4.6 mm * 100 mm 5um, 100A, Selectra C18 Guard Cartridges SLC-18GDC46-5UM, separation temperature 35 °С, flow rate 0.3 ml/min, injected sample volume - 2 μl, analysis time - 10 min. Used a gradient elution: the ratio of the solution of 0.1 % formic acid and acetonitrile was at 0 min 75 and 25 %; 0.4 min 60 and 40 %; 6 minutes 20 and 80 %; 8 minutes 75 and 25 %. Under these conditions, the retention time of methotrexate is 3.11 minutes. Detection conditions: methotrexate - positive ionization mode, 455.15 m / z → 308.125 m / z, collision energy 22.99 V, source fragmentation 5, CID gas pressure 2 mTorr. The extraction of methotrexate from the transport medium (Hanks solution with 25 mM Hepes and 1% dimethyl sulfoxide) after incubation with Caco-2 cells for 3 h was carried out with a mixture of methanol + water in a ratio of 1: 1. Results. The developed method was validated according to the following parameters: selectivity, linearity, accuracy, precision, limit of quantitative determination, sample transfer, sample stability. The confirmed analytical range of the method was 60 -10,000 nmol / L in the transport medium. Conclusions: a method for the quantitative determination of methotrexate in the transport medium of Caco-2 cells by HPLC-MS / MS was developed and validated.

Effect of chemical modification of hydrodistillation on yield, composition and biological activity of Rosmarinus officinalis essential oil

Studies on the chemical modifications of Rosmarinus officinalis essential oil hydrodistillation process (HD) by using 5% citric acid (CA-HD) and 5% trisodium citrate (TSC-HD) as a water phase were performed. Composition of essential oils obtained in conventional and modified conditions was analyzed by gas chromatography with mass selective detector method (GC-MS) and compared. Antioxidant activity of all essential oils was determined spectrophotometrically by using DPPH radical scavenging method. It was found that applied modifications of hydrodistillation process enhanced yields and antioxidant activity and the best results were obtained using 5% citric acid as a modifier. Effect of this modification on fungicidal activity of essential oils against 8 various fungi strains (Alternaria alternata, Botrytis cinerea, Fusarium culmorum, Phythophtora cactorum, Rhizoctonia solani, Phythophtora infestans, Sclerotinia sclerotiorum and Ascosphaera apis) was also determined and in most cases enhanced activity was observed.

Gas chromatography-Mass spectrometry (GC-MS) analysis of bioactive components present in grape citrus peel in Nigeria

Grapefruit (Citrus paradisi) is an important cultivar of the Citrus genus which contains a number of nutrients beneficial to human health. Grapefruit peels are usually thrown away in this part of Nigeria. The research work investigated the bioactive components present in an indigenous citrus peel, grape (Citrus paradisi). Grape fruits were purchased from the fruits garden market in D-Line, Port-Harcourt metropolis and washed with ionized water and allowed to shade dry. The peel of the fruits were separated and subjected to cold extraction using 95% ethanol. The extracts obtained were further extracted in dichloromethane and subjected to GC/MS analysis for characterization of various bioactive components. The gas chromatographic model: 789A (GC) analysis was performed on an agilent technologies interfaced with mass selective detector model: 5975(MSD). The results revealed 25 bioactive components in grape peel with n-Hexadecanoic acid showing the highest concentration of 20.36% and retention time of 18.522min. Nootkatone was the lowest component in the grape peel with concentration 0.74% and retention time of 16.459min. Results shows that grapefruit (Citrus paradisi) has considerable potential as a source of natural bioactive components with different retention times. These fruits residues which otherwise regarded as waste hold promising potentials for medicinal therapy and value added food supplements.

Sonication of Petrochemical Industry Wastewaters

The objective of the study was the treatment of the pollutants (dissolved chemical oxygen demand (CODdis), total organic carbon (TOC) and total and individual polycyclic aromatic hydrocarbons (PAH)) present in the petrochemical industry wastewater (PCI) by sonication. The effects of increasing sonication times (0 min, 60 min, 120 and 150 min), sonication temperatures (25oC, 30oC and 60oC), on the CODdis, TOC and (PAH) removal efficiencies were researched at a sonication frequency of 35 kHz and a sonication power of 640. All the PAHs and their metabolites were measured by an gas chromatography (Agilent 6890 NC) equipped with a mass selective detector (Agilent 5973 inert MSD) with a capillary column (HP5-MS, 30 m, 0.25mm, 0.25µm)). The CH4, CO2 and H2 gas analysis, CODdis, TOC and the other pollutants were measured according to Standard Methods. As the sonication time and temperature were increased from 60 to 120 and 150 min, and from 25oC to 30oC and to 60oC, the CODdis, total PAH and TOC yields increased from 80.16% to 92.15%, from 78.37% to 94.23% and from 79.65% to 96.90%, respectively. The PAHs intermediates namely, 1–methylnaphthalene, 9–hydroxyfluorene, 9,10–phenanthrenequione, benzoic acid, 1,2,3–thiadiazole–4–carboxylic acid, naphthalene, p–hydroxybenzoic acid, fluorene, di–hydroxy pyrene, pyrene di–hydrodiol were sonodegraded with yields of 92.11%, 95.23%, 98.42%, 97.34%, 99.44%, 96.30%, 99.36%, 97.17%, 99.63% and 99.98% respectively, after 150 min, at 25oC. The presence of CH4, H2 and CO2 gases during sonication showed that the degradation mechanism of the PAHs is ‘‘pyrolysis”.

In silico identification of compounds from Nigella sativa seed oil as potential inhibitors of SARS-CoV-2 targets

Background The growing number of cases, severity and fatality of the COVID-19 pandemic, coupled with the fact that no cure has been found has made infected individuals especially in Africa, to resort to the consumption of different natural products to alleviate their condition. One of such plant materials that have been consumed to remedy the severity of this viral infection is the oil of Nigella sativa seed commonly called black seed oil. In this study, we extracted and characterized the oil from this seed using gas chromatography coupled to a mass selective detector to identify the component phytochemicals. Site-directed multiligand docking of the identified compounds was performed on SARS-CoV-2 molecular targets- Replicase polyprotein 1a, RNA binding protein of NSP9, ADP ribose phosphatase of NSP3, 3-chymotrypsin-like protease 3CLpro, and RNA-dependent RNA polymerase RDRP, and ACE2–angiotensin-converting enzyme from the Homo sapiens. Results The binding affinity of caryophyllene oxide was the highest on 3CLpro (− 6.0 kcal/mol), NSP3 (− 6.3 kcal/mol), NSP9 (− 6.3 kcal/mol), and RDRP (− 6.9 kcal/mol) targets, while α-bergamotene gave the best binding affinity on RPIA (5.7 kcal/mol) target. The binding affinity of β-bisabolene on the ACE2 target (− 8.0 kcal/mol) was almost the same as Remdesivir (− 8.1 kcal/mol). The ADMET properties of these three phytochemicals showed that they are good drug leads for these SARS-CoV-2 receptors. Conclusion The findings from this study strongly indicate that the reported recovery from COVID-19 infection claimed by patients who consumed black seed oil could be linked to the presence of caryophyllene oxide, α-bergamotene, and β-bisabolene in this natural product.

Excretion of ropivacaine with breast milk during prolonged analgesia of a postoperative wound

The aimof our study was to evaluate the safety of ropivacaine given to lactating patients as a continuous infusion according to the selected local continuous wound infiltration (CWI) protocol after cesarean section (CS). Materials and methods.Elective CSs were performed under spinal anesthesia with bupivacaine 5% -2 ml at the L2L3 level. At the end of the operation, a 22 cm multiperforated catheter was placed in the surgical wound under the aponeurosis. 3 hours after spinal anesthesia a loading dose 8 ml of 0.2% ropivacaine was administered and followed by continuous infusion at rate 4 mL/h for 48 hours. Aside from CWI, all patients received systemic anesthesia: acetaminophen 4000 mg per os, ketorolac 90 mg IV and rescue opioid tramadol up to 400 mg (as-needed) daily. Breast milk (colostrum) samples were collected after 24 and 48 hours after a loading dose and were analyzed by gas chromatography with a mass selective detector for total ropivacaine concentrations. Results.The number of patients included in the study was 8. During the study neither cases of the onset of symptoms associated with systemic toxicity of ropivacaine, nor allergic and infectious complications, were observed. The mean +- SD Cmax total colostrum ropivacaine concentration was 0.005 0.002 (0.002 0.007) g / ml. Conclusion.In this limited sample, increasing the concentration or/and infusion rate seems to be a safe (for both mother and newborn) alternative or adjunct to standard systemic analgesia after CS in order to provide proper postoperative pain control.

EXPERT STUDY OF ISONIAZIDE IN MEAT PRODUCTS

The article deals with the recommendations for the detection of isoniazid in meat products, using the example of minced meat using physicochemical methods. This study does not lose its relevance over the years, due to the use of drugs for tuberculosis for poisoning homeless and domestic animals, in particular dogs, in different regions of Ukraine, the main active ingredient of which is isoniazid. The article describes a method for the extraction of isoniazid in food products of meat origin (minced meat, sausage), which presumably could or may be used to poison domestic animals, in particular dogs. Isoniazid was identified by the methods of qualitative color reactions, thin-layer chromatography, and gas chromatography with a mass-selective detector. The process of qualitative identification of isoniazid in foodstuffs that could or may be used to poison domestic animals, in particular dogs, includes: -the method of extraction and separation of interfering impurities, primarily fats, preliminary research by the method of qualitative analytical reactions; -studies by thin layer chromatography; -research by instrumental methods, in particular, gas chromatography with mass-selective detection. The method for extracting isoniazid from meat products described in the article is effective and allows you to get rid of interfering substances, especially fats. In addition, the method of gas chromatography with mass-selective detection has sufficient sensitivity and selectivity. It allows with a minimum number of interfering peaks of low intensity, determine the composition of isoniazid accurately. The specified list of instrumental base, methods and research techniques is not exhaustive. The examiner should be guided by the availability of relevant literature and the availability of appropriate analytical equipment (gas chromatograph, gas chromatograph with mass-selective detector, liquid chromatograph, liquid chromatograph with mass-selective detector, etc.). In accordance with this, it is necessary to use certain methods for studying substances. These methods are subject to mandatory verification before being used in the laboratory. The obtained research results allow us to recommend this technique for implementation in expert practice.