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

SLOW QUASIKINETIC CHANGES IN WATER-LACTOSE COMPLEXES DURING STORAGE

Objective: To investigate kinetic changes in the spectral characteristics by Fourier Transform Infrared spectroscopy (FTIR) of water-lactose complexes (SMC), derived during the manufacturing process of the drug, containing release-active forms of antibodies. Methods: lactose monohydrate substance, saturated with release-active forms of affinity-purified polyclonal rabbit antibodies to recombinant human interferon-gamma (RA forms of Abs); tablets produced from this substance by direct compression after the addition of excipients (microcrystalline cellulose, magnesium stearate). Powdered and tableted placebo samples saturated with technologically processed water or phosphate-buffered saline, as well as with intact ethanol were used as control. Kinetic changes in SMC were studied using an Agilent Cary 630 FTIR spectrophotometer with a diamond ATR accessory (Agilent Technologies, USA). We used the method of X-ray fluorescence spectroscopy (EDX-7000 Shimadzu energy dispersive X-ray fluorescence spectrometer) to track changes in the fluorescence signal at certain wavelengths. The range of measured elements–11Na-92U. Results: Control of some technological characteristics of the obtained active substance (moisture, flowability) and dosage form (mean mass, disintegration rate) was used as indirect indicators of quality, but they did not allow reliably distinguishing intact lactose from the saturated one. Long-period oscillations on FTIR spectra were characteristic for all types of samples; oscillations occur at approximately two-week intervals; S/N indices were more stable for samples of RA forms of Abs than for placebo samples. On some days, the substance saturated with RA forms of Abs significantly differed from the intact lactose powder. The kinetics of the X-ray fluorescence intensity at certain wavelengths indicates the possibility of a periodic cooperative trigger transition of the system. Reversible conformational transitions are observed for powders on the 30th and 130th days (Kα 3.313 keV). For tablets at Kα 3.313 keV and Kα 1.740 keV small changes were visualized on those days (100–110th day) when hysteresis phenomena were recorded in the IR spectra of these samples. Conclusion: As a result, the evidence for a long-period dramatic conformational mobility of the water-lactose complex was obtained. Based on the data on the semiannual kinetics of IR spectra, a universal criterion for the identity of lactose powder saturated with RA forms of Abs was obtained. Also, it was confirmed that the lactose conformation state was changed by saturation with RA forms of Abs

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

Retrospective review of methodological foundations for the new monograph «Spectrometry in the near infrared regioN»

The new pharmacopoeial monograph «Spectrometry in the near infrared region» reflects the results of a difficult path traversed from the development of measurement and analytical techniques to the introduction of NIR spectroscopy in the practice of quality control. This review discusses the new monograph of the XIII edition of the State Pharmacopoeia in the light of existing methodological approaches developed and validated for the assessment of drug quality parameters such as identification, content uniformity, assay, etc. Special attention is paid to the formation of spectral libraries - the factor of the sample’s «age» is considered as a source of additional spectral variability. The review raises the question of the NIR spectra identity criteria. Drawing on the example of techniques developed for the evaluation of batch variability (23 medicines) it was showed that obtained values of spectral range serve as unique characteristics of each individual product and production process, defining interlot reproducibility - the less the spectral distance between the series is, the more homogeneous the products are. NIR spectroscopy was shown to be effective for quantitative analysis of solid dosage forms and liquid substances. The authors considered the principles of operation as well as prospects for the practical use of the new technology - NIR chemical imaging, combining spectral analysis and digital image

Mechanical Transformation of Compounds Leading to Physical, Chemical, and Biological Changes in Pharmaceutical Substances

This study demonstrates the link between the modification of the solid-phase pharmaceutical substances mechanical structure and their activity in waters with different molar ratio «deuterium-protium». Mechanochemical transformation of the powders of lactose monohydrate and sodium chloride as models of nutrients and components of dosage forms was investigated by the complex of physicochemical and biological methods. The solubility and kinetic activity of substances dispersed in various ways showed a positive correlation with the solvent isotope profile. Substances dissolved in heavy water were more active than solutes in natural water. Differential IR spectroscopy confirmed the modification of substituents in the sample of lactose monohydrate, demonstrating physicochemical changes during mechanical intervention. The biological activity of the compounds was determined by the method of Spirotox. The activation energy was determined by Arrhenius. Compared with the native compound, dispersed lactose monohydrate showed lower activation energy and, therefore, greater efficiency. In conclusion, proposed data confirm the statement that mechanical changes in compounds can lead to physicochemical changes that affect chemical and biological profiles

Ретроспективный обзор методологических основ новой ОФС «Спектрометрия в ближней инфракрасной области»

The new pharmacopoeial monograph «Spectrometry in the near infrared region» reflects the results of a difficult path traversed from the development of measurement and analytical techniques to the introduction of NIR spectroscopy in the practice of quality control. This review discusses the new monograph of the XIII edition of the State Pharmacopoeia in the light of existing methodological approaches developed and validated for the assessment of drug quality parameters such as identification, content uniformity, assay, etc. Special attention is paid to the formation of spectral libraries - the factor of the sample’s «age» is considered as a source of additional spectral variability. The review raises the question of the NIR spectra identity criteria. Drawing on the example of techniques developed for the evaluation of batch variability (23 medicines) it was showed that obtained values of spectral range serve as unique characteristics of each individual product and production process, defining interlot reproducibility - the less the spectral distance between the series is, the more homogeneous the products are. NIR spectroscopy was shown to be effective for quantitative analysis of solid dosage forms and liquid substances. The authors considered the principles of operation as well as prospects for the practical use of the new technology - NIR chemical imaging, combining spectral analysis and digital image.Новая фармакопейная статья «Спектрометрия в ближней инфракрасной области» отражает результаты трудного пути, пройденного от разработки измерительных и аналитических технологий, лежащих в основе метода, до внедрения БИК-спектроскопии в практику контроля качества лекарственных средств. В настоящем обзоре новая общая фармакопейная статья, вошедшая в XIII издание Государственной фармакопеи Российской Федерации, обсуждается в аспекте уже существующих методологических подходов, разработанных и валидированных для оценки таких показателей качества лекарственных средств, как подлинность, однородность дозирования, количественное определение и др. Отдельное внимание уделяется вопросам формирования спектральных библиотек - рассматривается фактор «возраста» образца как источника дополнительной вариабельности спектров. Поднимается вопрос критериев идентичности БИК-спектров лекарственных средств: на примере разработанных методик оценки вариабельности серийной продукции 23 наименований лекарственных препаратов показано, что полученные величины спектрального расстояния - уникальные характеристики каждого отдельного препарата и производственного процесса, определяющие посерийную воспроизводимость - чем меньше спектральное расстояние между сериями, тем однороднее продукция. Показаны возможности метода БИК-спектроскопии в количественном анализе как твердых лекарственных форм, так и жидких субстанций. Рассмотрен принцип работы, а также перспективы практического применения новой технологии БИК химической визуализации, сочетающей спектральный анализ и цифровое изображение

Controlling the Quality of Nanodrugs According to Their New Property—Radiothermal Emission

Previous studies have shown that complexly shaped nanoparticles (NPs) have their intrinsic radiothermal emission in the millimeter range. This article presents a method for controlling the quality of nanodrugs—immunobiological preparations (IBPs)—based on the detection of their intrinsic radiothermal emissions. The emissivity of interferon (IFN) medicals, determined without opening the primary package, is as follows (µW/m2): IFN-α2b—80 ± 9 (105 IU per package), IFN-β1a—40 ± 5 (24 × 106 IU per package), IFN-γ—30 ± 4 (105 IU per package). The emissivity of virus-like particles (VLP), determined using vaccines Gam-VLP-multivac (120 μg) in an injection bottle (crimp cap vials), was as follows: 12 ± 1 µW/m2, Gam-VLP—rota vaccines—9 ± 1 µW/m2. This study shows the reproducibility of emissivity over the course of a year, subject to the storage conditions of the immunobiological products. It has been shown that accelerated aging and a longer shelf life are accompanied by the coagulation of active NPs, and lead to a manyfold drop in emissivity. The dependence of radiothermal emission on temperature has a complex, non-monotonic nature. The emission intensity depends on the form of dosage, but remains within the order of magnitude for IFN-α2b for intranasal aqueous solution, ointments, and suppositories. The possibility of the remote quantitative control of the first phases of the immune response (increased synthesis of IFNs) to the intranasal administration of VLP vaccines has been demonstrated in experimental animals