PREPARATION, CHARACTERIZATION AND STUDIES OF PHYSICOCHEMICAL AND BIOLOGICAL PROPERTIES OF DRUGS COATING LACTOSE IN FLUIDIZED BEDS

Objective: Study physicochemical properties and activity of biotechnological drugs coating lactose particles in fluidized beds for the development of a prospective approach of their identification. Methods: Lactose monohydrate as pharmaceutical excipient; affinity-purified polyclonal rabbit antibodies to recombinant human interferon-gamma as a drug substance; Pilotlab fluid bed apparatus was used for lactose powder saturation with solutions of pharmaceutical substances to the point of granulation (pelletizing); inverse light scattering method (2D-LS) for analysis of micron vibrations frequency spectra of samples surfaces for light intensity distribution in time by values of d1, d2, d3 primary descriptors; low angel and dynamic laser light scattering (LALLS, DLS) methods for distribution of lactose-water (LW) supramolecular complexes into volume fractions (micron "size spectra"), using the Master Sizer 2000 instrument and Zeta Sizer Nano ZS instrument in the nanoscale; Spirotox method for research of biological activity to determine the activation energy (Ea) values of cell death in solutions of tested samples. Results: Changes in 2D-LS scattering time on sample surfaces, described by topological descriptors, made it possible to clearly differentiate the intact lactose from fluidized samples by specific corridors in coordinates di=F(t). The calculated activation energy (Ea) values of cell biosensor death process in solutions of drugs coating lactose allow to characterize the biological activity of it in the initial (by Ea increase) and activated state (by Ea decrease) after the creation of intra-laboratory transmucosal conditions. A unique dimensional spectrum of LW complexes in the nanoscale range was obtained by DLS. The differences between samples in the distribution of LW complexes in the size range from 1 µm to 125 µm was showed by LALLS. Conclusion: The developed approach, including Сhemometrics, laser and biotesting methods can be used for qualitative the analysis tasks as well as for analytical control of the fluidization process in cases where identifiable pharmaceutical substances are not distinguishable by traditional analytical methods

ASSESSMENT OF BIOLOGY ACTIVITY OF THE PEELING SUBSTANCES BY THE PHYSICOCHEMICAL APPROACHES ON THE SPIROSTOMUM AMBIGUUM CELL MODEL

Objective: To evaluate the biological activity of chemical peeling substances based on enzymatic and Arrhenius kinetics using Spirostomum ambiguum as an alternative approach to animal experiments. Methods: The Spirotox method was used to analyze the mechanism of «xenobiotic-cell» interaction, similar to the Michaelis-Menten enzymatic kinetics. The Hill-Langmuir equation was used to determine the degree of cooperativity in the binding of xenobiotics to cellular receptors. Using the Arrhenius kinetics, the observed activation energy obsEa of cell death in the model solutions of glycolic and carbolic acids was determed, which will allow predicting the toxicity parameters of any peeling substances. Results: The relationship Spirostomum ambiguum lifetime tL-lgC concentration of peeling compound solution made it possible to characterize the moment of cellular transition from the intermediate state C•Ln to the dead state DC, characterized by irreversible structural and functional changes in the cell/death. The values were 5.3 mmol•l-1 for glycolic acid solutions and 2.8 mmol•l-1 for carbolic acid solutions. Equilibrium constants Keq of complexation, the rate of infusoria death fm, and the degree of ligand cooperativity n were calculated. The activation energy °bsEa of cell death was determined in Arrhenius coordinates, which were 210±0.39 kJ·mol-1 and 108±0.09 kJ·mol-1 for glycolic and carbolic acids respectively. The correlation between the values ​​of activation energy and DL50 of mammals (rats) was discovered. Conclusion: The obtained kinetic parameters made it possible, without animals and humans testing, to characterize the mechanisms of interaction of peeling substances with the living cell

LIGHT SCATTERING IN RESEARCH AND QUALITY CONTROL OF DEUTERIUM DEPLETED WATER FOR PHARMACEUTICAL APPLICATION

Objective: Development of a methodology for measuring the deuterium content in water for pharmaceutical purposes by laser light scattering based on ideas about the cluster structure of water. Methods: Samples of industrially manufactured drinking water from different manufacturers with varying deuterium content from 10 ppm to 115 ppm. For the titration of laboratory samples of deuterium depleted water in increments of 5 ppm the following reagents were used: Water, deuterium-depleted (≤1 ppm (D2O, Aldrich, USA); Deuterium oxide/Heavy water/Water-d2 (99.9 atom % D, Aldrich, USA); water Milli-Q (specific resistance 18.2 µS·sm at 25 оС, ТОС ≤ 5 ppb, Merck Millipore). The determination of deuterium content in samples of industrially manufactured water and water obtained in a laboratory manner was carried out by the method of low-angle laser light scattering (LALLS) at the Mastersizer (Malvern Instruments) analyzer and using a working measuring tool–laser dispersion meter/MDL («Cluster-1», Russia/Ukraine). The statistical methods–packages OriginPro®9. Results: It was found that the content of isotopologies in water leads to physicochemical water’s properties changes and morphology changes of giant heterogeneous clusters (GHC). The results of low-angle laser light scattering (LALLS) in the water samples under investigation showed the dependence of the water GHC "dispersibility" expressed in the differentiation of curves of the volume size distribution ("size spectra"), the volume concentration, w%, the laser obscuration values (I ‒I0) as the function of the water isotopic composition variations. The laser diffraction method results correlate with two-dimensional (2D) multi-descriptor mathematical analysis. Conclusion: When identifying deuterium depleted water, it should be considered not only the indicators that determine its pharmacopoeial quality, but also the D/H ratio, because even small changes in the natural isotopic composition of water lead to significant biological effects. Our proposed approach using laser diffraction in combination with mathematical apparatus of (2D) multi-descriptor laser scattering analysis makes possible the exact calculation of individual signs of deuterium depleted water as the pharmaceutical object of study

Sorption Properties of Clay and Pectin-Containing Hydrogels

As is known, polymeric polyelectrolyte hydrogels are superabsorbents that are capable of absorbing moisture in amounts many times greater than their own mass. Numerous studies have shown that besides water absorption and retention, they can also be effectively used as sorbents to purify water from heavy metals. In many works, attempts are made to improve the sorption properties of polyelectrolyte hydrogels by creating polymer composites based on them. Organic/inorganic composite materials frequently exhibited desired hybrid performance superior to their individual components and cost-efficient characteristics. The composites derived from natural polysaccharides and inorganic clay minerals are of special interest by virtue of their unique commercial and environmental advantages, which means that the design and development of environmentally friendly superabsorbents, introducing natural ingredients, have long been necessary. In this paper, we consider polymer hydrogels based on a copolymer of acrylic acid and acrylamide filled with pectin and bentonite. The aim of this study is to investigate the influence of chemical conditions on hydrogels and their composites, kinetic, and absorption behavior toward metal ions in the presence of the chelating agent. In this chapter, an investigation of the kinetic patterns of swelling, deswelling, and sorption of the hydrogels and their composites will be presented

ASCORBIС ACID DEGRADATION IN N, N-DIMETHYLFORMAMIDE SOLUTIONS

Objective: Investigate the mechanisms of L-ascorbic acid transforтmation and formation of coloured enamines in N, N-dimethyl-formamide solutions. Methods: An automatic polarimeter Atago POL-1/2 was used for polarimetric investigation. Electronic spectra were recorded by UV-spectrometer Cary 60 (Agilent). The statistical analysis was carried out using the OriginPro 9.1 packages. Results: The Biot’s law violation was found in below 0.1% solutions of L-ascorbic acid (AA) in N, N-dimethylformamide (DMF). During the day, the specific rotation   of 1% AA solution varied from+37 to-1.0. Gradually, the solution acquired the red colour, and its intensity depended on the AA concentration. Spectrophotometrically, it was shown that after 15 min AA was absent in the n·10-3% solutions. The decomposition followed the first-order kinetics (k1=1.83·10-2с-1). At the same time, new absorption bands appeared at 273, 390, 533 nm. Model solutions containing dimethylamine (DMA) had a similar spectrum, and the intensity of the absorption bands increased in proportion to the concentration of DMA. Conclusion: The results show that the first step in the decomposition of ascorbic acid AA in DMF follows first-order kinetics. Numerous decomposition products are optically active compounds and reverse the sign of the optical rotation of the solution. The water resulting from the decomposition of AA is involved in the hydrolysis of the solvent. The hydrolysis product, the secondary amine DMA, interacts with the carbonyl groups of the AA decomposition products to form coloured enamines. Magnesium (II) accelerates the formation of coloured products

PHYSICOCHEMICAL PROPERTIES AND BIOLOGICAL ACTIVITY OF THE NEW ANTIVIRAL SUBSTANCE

Objective: To develop a set of quality control procedures for the promising antiviral pharmaceutical substance L-histidyl-1-adamantylethylamine dihydrochloride monohydrate, a derivative of rimantadine. Methods: Substances and solvents: synthesized in laboratory L-histidyl-1-adamantylethylamine dihydrochloride monohydrate (H-His-Rim•2HCl•H2O), rimantadine hydrochloride (Rim•HCl), 99%, ethanol 96%, N, N-dimethylformamide (DMF) anhydrous, 99.8% and n-hexane anhydrous, 95%, deionized high-resistance water (18.2 MΩ•cm at 25 °C, Milli-Q system), silver nitrate. Infrared (IR) Spectroscopy–Cary 630 Fourier Transform IR Spectrometer, elemental analysis–elemental composition analyzer CHNS-O EuroEA3000, ultraviolet (UV) spectrometry–Cary-60 spectrophotometer, polarimetry–POL-1/2 polarimeter with an external Peltier module, granulometric analysis by optical microscopy (Altami BIO 2 microscope) and low-angle laser light scattering (LALLS)–Master Sizer 3600, measurement of potential for hydrogen–potentiometer PB-11, Spirotox method–the study of temperature dependences of Spirostomum ambiguum lifetime to characterize the biological activity of the studied compounds. Results: The substance H-His-Rim•2HCl•H2O is an amorphous yellowish powder, slightly soluble in water, soluble in ethanol, freely soluble in N, N-dimethylformamide, and practically insoluble in n-hexane. A study of the elemental composition has confirmed the authenticity of H-His-Rim•2HCl•H2O. Comparison of the spectral characteristics of H-His-Rim•2HCl•H2O and Rim•HCl by IR spectroscopy and UV spectrometry confirmed the authenticity of the substance. The racemic form of the substance Rim•HCl with an insignificant amount of impurity of the levorotatory enantiomer was proved polarimetrically: α =-0.0126±0.0003 (1% aqueous solution, 20±0.5 °С). The specific optical rotation of 1% aqueous solution H-His-Rim•2HCl•H2O . In 1% ethanol solution -10.32±0.12. Using the method of laser light diffraction for a substance H-His-Rim•2HCl•H2O, the dimensional spectra «fraction of particles, %-d, μm» were characterized, the maximum of which in hexane is in the region of 40–50 μm. Arrhenius’s kinetics on the Spirotox model established statistically significant differences in ligand-receptor interactions, which are characterized by values of observed apparent activation energy °bsEa, kJ/mol: 132.36±1.55 for H-His-Rim•2HCl•H2O and 176.15±0.48 for Rim•HCl. Conclusion: The developed set of methods for assessment of physical and chemical properties and biological activity of a new antiviral substance H-His-Rim•2HCl•H2O is the basis for establish of regulatory documentation

POLARIMETRIC RESEARCH OF PHARMACEUTICAL SUBSTANCES IN AQUEOUS SOLUTIONS WITH DIFFERENT WATER ISOTOPOLOGUES RATIO

Objective: Methodology development for quality control of optically active pharmaceutical substances based on water isotopologues. Methods: Solutions of L-ascorbic acid, glucose, galactose and valine stereoisomers were prepared using deuterium depleted water (DDW-«light» water, D/H=4 ppm), natural deionized high-ohmic water (BD, D/H=140 ppm), heavy water (99.9% D2O). The optical rotation was observed using an automatic polarimeter Atago POL-1/2. The size distribution of giant heterogeneous clusters (GHC) of water was recorded by low angle laser light scattering (LALLS) method. Results: The infringement of Biot's Law was found for solutions of ascorbic acid, expressed in the absence of a constant value of the specific optical rotation  at a concentration of below 0.1%, depends on the D/H ratio. The inequality was established in absolute values of optical rotation for L-and D-isomers of valine in solutions with different ratios of hydrogen isotopologues. The mutarotation of glucose confirmed the first-order kinetics, and the activation energies were statistically distinguishable for BD and DDW. The mutarotation of the natural galactose D-isomer proceeded with a lower energy consumption compared to the L-isomer. In heavy water, the mutarotation of monosaccharides had different kinetic mechanisms. Polarimetric results correlated with the number and size of GHC, which confirmed the possibility of chiral solvent structures induction by optically active pharmaceutical substances. Conclusion: In the optically active pharmaceutical substances quality control there should be considered the contribution of induced chiral GHC of water to the optical rotation value that depends on the isotopic D/H ratio, the substance nature and the form of its existence at a given pH

pH-чувствительные съедобные пленки на основе биокомпозита казеинат натрия/агар-агар

In the modern world, shelf life extension of foods with a possibility of controlling their freshness and quality in real time is gaining increasing importance. To solve this task, the development of pH-sensitive films based on edible biopolymers with addition of anthocyans as a color indicator can be proposed. For this experiment, sodium caseinate/ agar-agar biocomposite films with different content of anthocyans from black carrot (Scorconera hispanica) in a range of 0 to 15 mass% in increments of 5 mass% were prepared. It was established that the anthocyan content did not affect the thickness and moisture content of the films; however, it significantly reduced their moisture absorption and transparence. Water vapor permeability of the biocomposite films increased with an increase in the content of the aqueous extract of anthocyans from black carrot, which can be linked with the formation of pores and microcracks in the biocomposite matrix. All films with anthocyans showed changes in color depending on the environmental pH, which intensities depended on the anthocyan content. Edible films based on the mixture of sodium caseinate and agar-agar can potentially be used in the food industry to determine freshness of foods, which spoilage results in pH changes.В современном мире все большее значение приобретает продление срока годности продуктов питания с возможностью контроля их свежести и качества в режиме реального времени. Для решения этой задачи может быть предложена разработка рН-чувствительных пленок на основе съедобных биополимеров с добавкой антоцианов в качестве цветового индикатора. Для проведения данного исследования были приготовлены пленки биокомпозита казеинат натрия/агар-агар с различным содержанием антоцианов черной моркови (Scorconera hispanica) 0–15 масс.% с шагом 5 масс.%. Было установлено, что содержание антоцианов не влияет на толщину и влагосодержание пленок, однако значительно снижает их влагопоглощение и прозрачность. Паропроницаемость пленок биокомпозита возросла с увеличением содержания водного экстракта антоцианов черной моркови, что может быть связано с образованием пор и микротрещин в матрице биокомпозита. Все пленки с антоцианами показали изменение цвета в зависимости от рН среды, интенсивность которого зависела от содержания антоцианов. Съедобные пленки на основе смеси казеината натрия и агар-агара потенциально могут быть использованы в пищевой промышленности для определения свежести продуктов, в результате порчи которых происходит изменение рН

Перинатальные факторы программирования девиаций физического развития преждевременно родившихся младенцев

The analysis of obstetric anamnesis and indicators of heart rate variability in the neonatal period in prematurely born children who had deviations of somatometric parameters from reference values at the age of 12 months was carried out. Excessive due to various reasons, the level of adrenomimetics supplied with maternal blood caused an increase in the intensity of metabolism in the fetal body and inhibition of the activity of sympathoadrenal systems. The insufficient provision of the fetal and newborn organism with energy and plastic materials that arose under these conditions initiated perinatal metabolic programming of deviations in the physical development of prematurely born infants.Материалы и методы. Проводился анализ акушерского анамнеза и показателей вариабельности сердечного ритма в неонатальном периоде у преждевременно родившихся детей, имевших отклонения соматометрических показателей от референсных значений в возрасте 12 месяцев.Результаты. Избыточный вследствие различных причин уровень адреномиметиков, поступавших с материнской кровью, вызывал в организме плода повышение интенсивности метаболизма и ингибирование активности симпатоадреналовых систем. Возникавшее в этих условиях недостаточное обеспечение организма плода и новорожденного энергетическими и пластическими материалами инициировало перинатальное метаболическое программирование девиаций физического развития преждевременно родившихся младенцев

Pulsed electron-nuclear double resonance diagnostics of Ce³⁺ emitters in scintillating garnets

© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, WeinheimPulsed electron paramagnetic resonance (EPR) and pulsed electron-nuclear double resonance (ENDOR) techniques have been applied to study the environment of luminescent Ce3+ ions in garnet based scintillator powders and ceramics. The presence of aluminum and gallium isotopes with large nuclear magnetic and quadrupole moments in the nearest neighborhood of the Ce3+ ion allows for the use of the hyperfine and quadrupole interactions with these ions for determination of the unpaired electron spatial distribution and the definition of the electric field gradient at the aluminum and gallium sites. Pulsed EPR and ENDOR techniques made it possible to study the coherent properties of the Ce3+ spin system in garnet powders and ceramics, which is important for spin manipulation on Ce3+ centers