12 research outputs found
Comparative Assessment of Quality Requirements for Medicinal Products Containing Diosmin
Currently, there is an increase in preand post-approval testing of medicinal products containing diosmin and hence a need to unify approaches to standardisation of this group of pharmaceuticals. Moreover, the State Pharmacopoeia of the Russian Federation lacks a monograph for these products.The aim of the study was to determine an approach to standardisation of medicinal products containing diosmin.Materials and methods: the study analysed scientific publications, as well as monographs of leading foreign pharmacopoeias. Experimental work was carried out using samples of diosmin-containing pharmaceuticals in the form of 500 and 1000 mg film-coated tablets produced by Russian and foreign manufacturers. The study involved high performance liquid chromatography with UV detection using an Agilent 1260 Infinity II liquid chromatography system with a diode array detector. The following reference standards were used: a diosmin RS, USP grade; a hesperidin CRS, Ph. Eur. Grade; and a diosmin CRS for testing chromatography system suitability for identification of impurities A, B, C, D, E, and F, Ph. Eur. grade.Results: the authors reviewed quality requirements for pharmaceutical products containing diosmin and analysed experimental data obtained during preand post-approval testing of Russian and foreign medicines. The comparison of regulatory documents for registered diosmin-containing medicinal products showed a difference in approaches to assessing the contents of related substances and active pharmaceutical ingredients. Having analysed the literature, experimental data and regulatory requirements for standardisation of diosmin-containing pharmaceuticals, the authors recommended an approach to standardisation. According to the approach, concomitant flavonoids (hesperidin, isorchoifolin, linarin, and diosmetin) contributing to the pharmacological activity of a medicinal product are specified as part of Assay, and process-related by-products (impurities A and D) are specified and evaluated as part of Related substances tests.Conclusion: the authors propose to evaluate the contents of concomitant flavonoids (hesperidin, isorchoifolin, linarin, diosmetin) under Assay and to specify impurities A and D, as well as single unidentified impurities and total amount of impurities under Related substances
DIAGNOSTIC POTENTIAL OF THE ERYTHROCYTIC IMMUNOGLOBULIN DIAGNOSTICUM FOR INDICATION AND IDENTIFICATION OF THE CAUSATIVE AGENTS OF PARTICULARLY DANGEROUS (DEEP) MYCOSES
Objective of the study was to assess analytical and diagnostic sensitivity and specificity of the βReagent kit. Erythrocytic coccidioidomycosal and histoplasmosal immunoglobulin dry diagnosticumβ, designed for identification of causative agents of coccidioidomycosis and histoplasmosis in isolated cultures of micromycetes, as well as in clinical and biological samples using indirect hemagglutination test.Materials and methods. The investigation included 264 positive samples (216 samples of micromycete suspensions, 48 samples of biological and clinical material) containing pathogens of histoplasmosis and coccidioidomycosis concentrated up to 3,12Β·106 and 1,56Β·106 cells/ml, respectively, and 128 negative samples containing heterologous microorganisms in concentrations equal to 5Β·106 cells/ml. The study was carried out using biological samples that were artificially contaminated with stated pathogens of particularly dangerous mycoses and samples, obtained from bioassay animals with experimental infection.Results and conclusions. It is established that diagnostic sensitivity of the reagent kit is not less than 99,0 %. The diagnostic specificity is not less than 98,0 %. Reproducibility of the results in all cases was 100 %. The results obtained testify to the prospect of introduction of the developed kit into the health care practice
Modern Approaches for Detection of Glanders and Melioidosis. Identification and Typing of <i>Burkholderia mallei</i> and <i>Burkholderia pseudomallei</i>
Analysed are the methodological approaches used for identification and typing of B. mallei and B. pseudomallei. Suggested are recommendations for improvements of algorithms of laboratory diagnosis of glanders and melioidosis including wide range of biochemical, immunodiagnostic and molecular genetic methods
ΠΡΠ΅Π½ΠΊΠ° Π΄ΠΈΠ°Π³Π½ΠΎΡΡΠΈΡΠ΅ΡΠΊΠΎΠΉ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ Π½Π°Π±ΠΎΡΠ° ΡΠ΅Π°Π³Π΅Π½ΡΠΎΠ² Π΄Π»Ρ in vitro Π΄ΠΈΠ°Π³Π½ΠΎΡΡΠΈΠΊΠΈ Π»ΠΈΡ ΠΎΡΠ°Π΄ΠΊΠΈ ΠΠ°ΠΏΠ°Π΄Π½ΠΎΠ³ΠΎ ΠΠΈΠ»Π° ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΏΠΎΠ»ΠΈΠΌΠ΅ΡΠ°Π·Π½ΠΎΠΉ ΡΠ΅ΠΏΠ½ΠΎΠΉ ΡΠ΅Π°ΠΊΡΠΈΠΈ Ρ ΠΎΠ±ΡΠ°ΡΠ½ΠΎΠΉ ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΈΠ΅ΠΉ ΠΈ Π³ΠΈΠ±ΡΠΈΠ΄ΠΈΠ·Π°ΡΠΈΠΎΠ½Π½ΠΎ-ΡΠ»ΡΠΎΡΠ΅ΡΡΠ΅Π½ΡΠ½ΠΎΠΉ Π΄Π΅ΡΠ΅ΠΊΡΠΈΠ΅ΠΉ
West Nile fever is a vector-borne zoonotic arbovirus infection with natural foci. Its clinical course is similar to that of acute febrile syndrome, and severe cases may result in neuroinvasive disease. Several genetic lineages (1, 2, and 4) of the West Nile virus (WNV) with different pathogenicity for humans are circulating in the Russian Federation. Therefore, it is an urgent task to develop a diagnostic reagent kit for differentiating between WNV genetic lineages and to implement the kit in clinical laboratory practice.The aim of the study was to conduct technical and clinical tests and evaluate the quality, efficacy, and safety of the Ampligen-WNV-genotype-1/2/4 diagnostic reagent kit for detecting WNV RNA and differentiating between WNV genetic lineages 1, 2, and 4 by reverse transcription polymerase chain reaction (RT-PCR) with fluorescent probe-based detection.Materials and methods. The authors determined the diagnostic sensitivity and specificity of the Ampligen-WNV-genotype-1/2/4 reagent kit (Volgograd Research Institute for Plague Control, Russia) by real-time RT-PCR with 216 clinical samples and 204 biological samples. Sanger sequencing was used as a reference method. Statistical analysis of clinical test results was carried out in accordance with the Russian national standard for clinical laboratory tests (GOST R 53022.3-2008).Results. When tested with the Ampligen-WNV-genotype-1/2/4 reagent kit, real-time RT-PCR demonstrated the analytical sensitivity of 1Γ104 Β GEq/mL for the detection of WNV cDNA of genetic lineages 1, 2, and 4. The assessment of its analytical specificity showed no positive results for cDNA samples of heterologous viruses at a concentration of 1Γ106 Β GEq/mL. The diagnostic sensitivity with the reagent kit was at least 98.5%, and the diagnostic specificity was at least 99%, with 90% confidence levels for both parameters.Conclusions. The Ampligen-WNV-genotype-1/2/4 reagent kit can be recommended for use in clinical laboratory diagnostics to detect WNV RNA and differentiate between WNV genetic lineages 1, 2, and 4.ΠΠΈΡ
ΠΎΡΠ°Π΄ΠΊΠ° ΠΠ°ΠΏΠ°Π΄Π½ΠΎΠ³ΠΎ ΠΠΈΠ»Π° β Π·ΠΎΠΎΠ½ΠΎΠ·Π½Π°Ρ ΠΏΡΠΈΡΠΎΠ΄Π½ΠΎ-ΠΎΡΠ°Π³ΠΎΠ²Π°Ρ Π°ΡΠ±ΠΎΠ²ΠΈΡΡΡΠ½Π°Ρ ΠΈΠ½ΡΠ΅ΠΊΡΠΈΡ Ρ ΡΡΠ°Π½ΡΠΌΠΈΡΡΠΈΠ²Π½ΡΠΌ ΠΌΠ΅Ρ
Π°Π½ΠΈΠ·ΠΌΠΎΠΌ ΠΏΠ΅ΡΠ΅Π΄Π°ΡΠΈ Π²ΠΎΠ·Π±ΡΠ΄ΠΈΡΠ΅Π»Ρ. ΠΠ°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΠ΅ ΠΏΡΠΎΡΠ΅ΠΊΠ°Π΅Ρ Π² Π²ΠΈΠ΄Π΅ ΠΎΡΡΡΠΎΠ³ΠΎ Π»ΠΈΡ
ΠΎΡΠ°Π΄ΠΎΡΠ½ΠΎΠ³ΠΎ ΠΈΠ½ΡΠΎΠΊΡΠΈΠΊΠ°ΡΠΈΠΎΠ½Π½ΠΎΠ³ΠΎ ΡΠΈΠ½Π΄ΡΠΎΠΌΠ°, Π² ΡΡΠΆΠ΅Π»ΡΡ
ΡΠ»ΡΡΠ°ΡΡ
β Ρ ΡΠ°Π·Π²ΠΈΡΠΈΠ΅ΠΌ Π½Π΅ΠΉΡΠΎΠΈΠ½ΡΠ΅ΠΊΡΠΈΠΈ. ΠΡΠ΄Π΅Π»ΡΡΡ Π½Π΅ΡΠΊΠΎΠ»ΡΠΊΠΎ Π³Π΅Π½ΠΎΡΠΈΠΏΠΎΠ² (1, 2, 4) Π²ΠΈΡΡΡΠ° ΠΠ°ΠΏΠ°Π΄Π½ΠΎΠ³ΠΎ ΠΠΈΠ»Π°, ΠΠΠ (West Nile virus, WNV), ΡΠΈΡΠΊΡΠ»ΠΈΡΡΡΡΠΈΡ
Π½Π° ΡΠ΅ΡΡΠΈΡΠΎΡΠΈΠΈ Π ΠΎΡΡΠΈΠΉΡΠΊΠΎΠΉ Π€Π΅Π΄Π΅ΡΠ°ΡΠΈΠΈ ΠΈ ΠΈΠΌΠ΅ΡΡΠΈΡ
ΡΠ°Π·Π½ΡΡ ΠΏΠ°ΡΠΎΠ³Π΅Π½Π½ΠΎΡΡΡ Π΄Π»Ρ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ°. Π ΡΠ²ΡΠ·ΠΈ Ρ ΡΡΠΈΠΌ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠΊΠ° ΠΈ Π²Π½Π΅Π΄ΡΠ΅Π½ΠΈΠ΅ Π² ΠΊΠ»ΠΈΠ½ΠΈΡΠ΅ΡΠΊΡΡ Π»Π°Π±ΠΎΡΠ°ΡΠΎΡΠ½ΡΡ ΠΏΡΠ°ΠΊΡΠΈΠΊΡ Π΄ΠΈΠ°Π³Π½ΠΎΡΡΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ Π½Π°Π±ΠΎΡΠ° ΡΠ΅Π°Π³Π΅Π½ΡΠΎΠ² Π΄Π»Ρ Π΄ΠΈΡΡΠ΅ΡΠ΅Π½ΡΠΈΠ°ΡΠΈΠΈ Π³Π΅Π½ΠΎΡΠΈΠΏΠΎΠ² ΠΠΠ ΡΠ²Π»ΡΠ΅ΡΡΡ Π°ΠΊΡΡΠ°Π»ΡΠ½ΠΎΠΉ Π·Π°Π΄Π°ΡΠ΅ΠΉ.Π¦Π΅Π»Ρ ΡΠ°Π±ΠΎΡΡ: ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½ΠΈΠ΅ ΡΠ΅Ρ
Π½ΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΈ ΠΊΠ»ΠΈΠ½ΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΈΡΠΏΡΡΠ°Π½ΠΈΠΉ Π΄Π»Ρ ΠΎΡΠ΅Π½ΠΊΠΈ ΠΊΠ°ΡΠ΅ΡΡΠ²Π°, ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΠΈ Π±Π΅Π·ΠΎΠΏΠ°ΡΠ½ΠΎΡΡΠΈ Π΄ΠΈΠ°Π³Π½ΠΎΡΡΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ Π½Π°Π±ΠΎΡΠ° ΡΠ΅Π°Π³Π΅Π½ΡΠΎΠ² Β«ΠΠΌΠΏΠ»ΠΈΠ³Π΅Π½-WNV-Π³Π΅Π½ΠΎΡΠΈΠΏ-1/2/4Β» Π΄Π»Ρ Π²ΡΡΠ²Π»Π΅Π½ΠΈΡ Π ΠΠ ΠΈ Π΄ΠΈΡΡΠ΅ΡΠ΅Π½ΡΠΈΠ°ΡΠΈΠΈ Π³Π΅Π½ΠΎΡΠΈΠΏΠΎΠ² (1, 2, 4) Π²ΠΈΡΡΡΠ° ΠΠ°ΠΏΠ°Π΄Π½ΠΎΠ³ΠΎ ΠΠΈΠ»Π° ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΏΠΎΠ»ΠΈΠΌΠ΅ΡΠ°Π·Π½ΠΎΠΉ ΡΠ΅ΠΏΠ½ΠΎΠΉ ΡΠ΅Π°ΠΊΡΠΈΠΈ Ρ ΠΎΠ±ΡΠ°ΡΠ½ΠΎΠΉ ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΈΠ΅ΠΉ ΠΈ Π³ΠΈΠ±ΡΠΈΠ΄ΠΈΠ·Π°ΡΠΈΠΎΠ½Π½ΠΎ-ΡΠ»ΡΠΎΡΠ΅ΡΡΠ΅Π½ΡΠ½ΠΎΠΉ Π΄Π΅ΡΠ΅ΠΊΡΠΈΠ΅ΠΉ.ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ: ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΠ΅ Π΄ΠΈΠ°Π³Π½ΠΎΡΡΠΈΡΠ΅ΡΠΊΠΎΠΉ ΡΡΠ²ΡΡΠ²ΠΈΡΠ΅Π»ΡΠ½ΠΎΡΡΠΈ ΠΈ ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ½ΠΎΡΡΠΈ Π½Π°Π±ΠΎΡΠ° ΡΠ΅Π°Π³Π΅Π½ΡΠΎΠ² Β«ΠΠΌΠΏΠ»ΠΈΠ³Π΅Π½-WNV-Π³Π΅Π½ΠΎΡΠΈΠΏ-1/2/4Β» (Π€ΠΠ£Π ΠΠΎΠ»Π³ΠΎΠ³ΡΠ°Π΄ΡΠΊΠΈΠΉ Π½Π°ΡΡΠ½ΠΎ-ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°ΡΠ΅Π»ΡΡΠΊΠΈΠΉ ΠΏΡΠΎΡΠΈΠ²ΠΎΡΡΠΌΠ½ΡΠΉ ΠΈΠ½ΡΡΠΈΡΡΡ Π ΠΎΡΠΏΠΎΡΡΠ΅Π±Π½Π°Π΄Π·ΠΎΡΠ°) ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΠ’-ΠΠ¦Π -Π Π Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ 216 ΠΎΠ±ΡΠ°Π·ΡΠΎΠ² ΠΊΠ»ΠΈΠ½ΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΠΈ 204 ΠΎΠ±ΡΠ°Π·ΡΠΎΠ² Π±ΠΈΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΠΌΠ°ΡΠ΅ΡΠΈΠ°Π»Π°. Π ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅ ΠΌΠ΅ΡΠΎΠ΄Π° ΡΡΠ°Π²Π½Π΅Π½ΠΈΡ ΠΏΡΠΈΠΌΠ΅Π½ΡΠ»ΠΈ ΡΠ΅ΠΊΠ²Π΅Π½ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΏΠΎ Π‘Π΅Π½Π³Π΅ΡΡ. Π‘ΡΠ°ΡΠΈΡΡΠΈΡΠ΅ΡΠΊΡΡ ΠΎΠ±ΡΠ°Π±ΠΎΡΠΊΡ ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΠΎΠ² ΠΊΠ»ΠΈΠ½ΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΈΡΠΏΡΡΠ°Π½ΠΈΠΉ ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ Π² ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²ΠΈΠΈ Ρ ΠΠΠ‘Π’ Π 53022.3-2008.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ: Π² Ρ
ΠΎΠ΄Π΅ ΠΈΡΠΏΡΡΠ°Π½ΠΈΠΉ ΡΡΡΠ°Π½ΠΎΠ²Π»Π΅Π½ΠΎ, ΡΡΠΎ Π°Π½Π°Π»ΠΈΡΠΈΡΠ΅ΡΠΊΠ°Ρ ΡΡΠ²ΡΡΠ²ΠΈΡΠ΅Π»ΡΠ½ΠΎΡΡΡ ΠΠ’-ΠΠ¦Π -Π Π c Π½Π°Π±ΠΎΡΠΎΠΌ ΡΠ΅Π°Π³Π΅Π½ΡΠΎΠ² Β«ΠΠΌΠΏΠ»ΠΈΠ³Π΅Π½-WNV-Π³Π΅Π½ΠΎΡΠΈΠΏ-1/2/4Β» ΡΠΎΡΡΠ°Π²ΠΈΠ»Π° 1Γ104 ΠΠ/ΠΌΠ» ΠΏΡΠΈ Π²ΡΡΠ²Π»Π΅Π½ΠΈΠΈ ΠΊΠΠΠ ΠΠΠ Π³Π΅Π½ΠΎΡΠΈΠΏΠΎΠ² 1, 2, 4. ΠΡΠΈ ΠΎΡΠ΅Π½ΠΊΠ΅ ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ½ΠΎΡΡΠΈ Π½Π°Π±ΠΎΡΠ° ΠΏΠΎΠ»ΠΎΠΆΠΈΡΠ΅Π»ΡΠ½ΡΡ
ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΠΎΠ² Ρ ΠΊΠΠΠ Π³Π΅ΡΠ΅ΡΠΎΠ»ΠΎΠ³ΠΈΡΠ½ΡΡ
Π²ΠΈΡΡΡΠΎΠ² Π² ΠΏΡΠΎΠ±Π°Ρ
Ρ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠ΅ΠΉ 1Γ106 ΠΠ/ΠΌΠ» Π½Π΅ Π²ΡΡΠ²Π»Π΅Π½ΠΎ. ΠΠΈΠ°Π³Π½ΠΎΡΡΠΈΡΠ΅ΡΠΊΠ°Ρ ΡΡΠ²ΡΡΠ²ΠΈΡΠ΅Π»ΡΠ½ΠΎΡΡΡ Π½Π°Π±ΠΎΡΠ° ΡΠ΅Π°Π³Π΅Π½ΡΠΎΠ² ΡΠΎΡΡΠ°Π²ΠΈΠ»Π° Π½Π΅ ΠΌΠ΅Π½Π΅Π΅ 98,5%, Π΄ΠΈΠ°Π³Π½ΠΎΡΡΠΈΡΠ΅ΡΠΊΠ°Ρ ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ½ΠΎΡΡΡ β Π½Π΅ ΠΌΠ΅Π½Π΅Π΅ 99%, ΠΏΡΠΈ Π΄ΠΎΠ²Π΅ΡΠΈΡΠ΅Π»ΡΠ½ΠΎΠΉ Π²Π΅ΡΠΎΡΡΠ½ΠΎΡΡΠΈ 90% ΠΏΡΠΈ Π°Π½Π°Π»ΠΈΠ·Π΅ ΠΊΠ°ΠΆΠ΄ΠΎΠ³ΠΎ ΠΈΠ· ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»Π΅ΠΉ.ΠΡΠ²ΠΎΠ΄Ρ: Π½Π°Π±ΠΎΡ ΡΠ΅Π°Π³Π΅Π½ΡΠΎΠ² Β«ΠΠΌΠΏΠ»ΠΈΠ³Π΅Π½-WNV-Π³Π΅Π½ΠΎΡΠΈΠΏ-1/2/4Β» ΠΌΠΎΠΆΠ΅Ρ Π±ΡΡΡ ΡΠ΅ΠΊΠΎΠΌΠ΅Π½Π΄ΠΎΠ²Π°Π½ Π΄Π»Ρ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΡ Π² ΠΊΠ»ΠΈΠ½ΠΈΡΠ΅ΡΠΊΠΎΠΉ Π»Π°Π±ΠΎΡΠ°ΡΠΎΡΠ½ΠΎΠΉ Π΄ΠΈΠ°Π³Π½ΠΎΡΡΠΈΠΊΠ΅ Π΄Π»Ρ ΠΎΠ±Π½Π°ΡΡΠΆΠ΅Π½ΠΈΡ Π ΠΠ ΠΈ Π΄ΠΈΡΡΠ΅ΡΠ΅Π½ΡΠΈΠ°ΡΠΈΠΈ Π³Π΅Π½ΠΎΡΠΈΠΏΠΎΠ² (1, 2, 4) Π²ΠΈΡΡΡΠ° ΠΠ°ΠΏΠ°Π΄Π½ΠΎΠ³ΠΎ ΠΠΈΠ»Π°
Metagenomic profiling of viral and microbial communities from the pox lesions of lumpy skin disease virus and sheeppox virus-infected hosts
IntroductionIt has been recognized that capripoxvirus infections have a strong cutaneous tropism with the manifestation of skin lesions in the form of nodules and scabs in the respective hosts, followed by necrosis and sloughing off. Considering that the skin microbiota is a complex community of commensal bacteria, fungi and viruses that are influenced by infections leading to pathological states, there is no evidence on how the skin microbiome is affected during capripoxvirus pathogenesis.MethodsIn this study, shotgun metagenomic sequencing was used to investigate the microbiome in pox lesions from hosts infected with lumpy skin disease virus and sheep pox virus.ResultsThe analysis revealed a high degree of variability in bacterial community structures across affected skin samples, indicating the importance of specific commensal microorganisms colonizing individual hosts. The most common and abundant bacteria found in scab samples were Fusobacterium necrophorum, Streptococcus dysgalactiae, Helcococcus ovis and Trueperella pyogenes, irrespective of host. Bacterial reads belonging to the genera Moraxella, Mannheimia, Corynebacterium, Staphylococcus and Micrococcus were identified.DiscussionThis study is the first to investigate capripox virus-associated changes in the skin microbiome using whole-genome metagenomic profiling. The findings will provide a basis for further investigation into capripoxvirus pathogenesis. In addition, this study highlights the challenge of selecting an optimal bioinformatics approach for the analysis of metagenomic data in clinical and veterinary practice. For example, direct classification of reads using a kmer-based algorithm resulted in a significant number of systematic false positives, which may be attributed to the peculiarities of the algorithm and database selection. On the contrary, the process of de novo assembly requires a large number of target reads from the symbiotic microbial community. In this work, the obtained sequencing data were processed by three different approaches, including direct classification of reads based on k-mers, mapping of reads to a marker gene database, and de novo assembly and binning of metagenomic contigs. The advantages and disadvantages of these techniques and their practicality in veterinary settings are discussed in relation to the results obtained
Π Π°Π·ΡΠ°Π±ΠΎΡΠΊΠ° ΡΠ½ΠΈΡΠΈΡΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΉ ΠΠΠΠ₯-ΠΌΠ΅ΡΠΎΠ΄ΠΈΠΊΠΈ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΡ ΡΠΎΠ΄ΡΡΠ²Π΅Π½Π½ΡΡ ΠΏΡΠΈΠΌΠ΅ΡΠ΅ΠΉ ΠΈ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠΉ ΠΎΡΠ΅Π½ΠΊΠΈ Π΄Π΅ΠΉΡΡΠ²ΡΡΡΠ΅Π³ΠΎ Π²Π΅ΡΠ΅ΡΡΠ²Π° ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ² ΠΏΠ°ΠΏΠ°Π²Π΅ΡΠΈΠ½Π° Π³ΠΈΠ΄ΡΠΎΡ Π»ΠΎΡΠΈΠ΄Π°
Abstract. Papaverine hydrochloride products are used as anticonvulsants in routine medical practice. Most of the approved product specification files include thin-layer chromatography for assessment of product-related impurities and UV spectrophotometry for determination of active pharmaceutical ingredients. An HPLC assay is not used for determination of papaverine hydrochloride in drug dosage forms.The aim of the study was to develop an HPLC test method for determination of product-related impurities and for quantification of papaverine hydrochloride in solutions for injection, tablets, and rectal suppositories.Materials and methods: samples of the following Russian-made papaverine products were used in the study: Papaverine, solution for injection, 20Β mg/mL; Papaverine, rectal suppositories, 20Β mg; Papaverine, tablets, 40Β mg. The Agilent 1260 Infinity II DAD System was used for the HPLC assay, and the Agilent 8453Π UV-Vis System was used for recording UV spectra. The determination of product-related impurities and the assay of active ingredients were performed simultaneously by HPLC using a reversed-phase column Kromasil 100-5-C18, 250Γ4.6Β mm, 5 ΞΌm, the gradient elution mode, and detection atΒ Β Β Β 238 nm. Papaverine Hydrochloride USP RS, 99% purity, and Noscapine EP CRS were used as reference standards.Results: the study demonstrated that determination of product-related impurities and assay of active ingredients in papaverine products can be performed simultaneously using HPLC.Conclusions: the authors proposed an HPLC test method for determination of active ingredients in papaverine products, which is aligned with the βconsistent standardisationβ principle and can be recommended for inclusion into draft monographs for papaverine products.Π Π΅Π·ΡΠΌΠ΅. ΠΡΠ΅ΠΏΠ°ΡΠ°ΡΡ ΠΏΠ°ΠΏΠ°Π²Π΅ΡΠΈΠ½Π° Π³ΠΈΠ΄ΡΠΎΡ
Π»ΠΎΡΠΈΠ΄Π° ΠΏΡΠΈΠΌΠ΅Π½ΡΡΡΡΡ Π² ΠΌΠ΅Π΄ΠΈΡΠΈΠ½ΡΠΊΠΎΠΉ ΠΏΡΠ°ΠΊΡΠΈΠΊΠ΅ Π² ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅ ΡΠΏΠ°Π·ΠΌΠΎΠ»ΠΈΡΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΡΡΠ΅Π΄ΡΡΠ²Π°. Π Π±ΠΎΠ»ΡΡΠΈΠ½ΡΡΠ²ΠΎ Π·Π°ΡΠ΅Π³ΠΈΡΡΡΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
Π½ΠΎΡΠΌΠ°ΡΠΈΠ²Π½ΡΡ
Π΄ΠΎΠΊΡΠΌΠ΅Π½ΡΠΎΠ² Π½Π° ΡΡΠΈ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΡ Π²ΠΊΠ»ΡΡΠ΅Π½Π° ΠΌΠ΅ΡΠΎΠ΄ΠΈΠΊΠ° ΠΎΡΠ΅Π½ΠΊΠΈ ΡΠΎΠ΄ΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΡΠΈΠΌΠ΅ΡΠ΅ΠΉ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΡΠΎΠ½ΠΊΠΎΡΠ»ΠΎΠΉΠ½ΠΎΠΉ Ρ
ΡΠΎΠΌΠ°ΡΠΎΠ³ΡΠ°ΡΠΈΠΈ, Π΄Π»Ρ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΡ Π΄Π΅ΠΉΡΡΠ²ΡΡΡΠ΅Π³ΠΎ Π²Π΅ΡΠ΅ΡΡΠ²Π° ΠΈΡΠΏΠΎΠ»ΡΠ·ΡΠ΅ΡΡΡ Π£Π€-ΡΠΏΠ΅ΠΊΡΡΠΎΡΠΎΡΠΎΠΌΠ΅ΡΡΠΈΡ. ΠΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠ΅ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΠ΅ ΠΏΠ°ΠΏΠ°Π²Π΅ΡΠΈΠ½Π° Π³ΠΈΠ΄ΡΠΎΡ
Π»ΠΎΡΠΈΠ΄Π° Π² Π»Π΅ΠΊΠ°ΡΡΡΠ²Π΅Π½Π½ΡΡ
ΡΠΎΡΠΌΠ°Ρ
ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ Π²ΡΡΠΎΠΊΠΎΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΠΉ ΠΆΠΈΠ΄ΠΊΠΎΡΡΠ½ΠΎΠΉ Ρ
ΡΠΎΠΌΠ°ΡΠΎΠ³ΡΠ°ΡΠΈΠΈ (ΠΠΠΠ₯) Π½Π΅ ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΡΡΡ.Π¦Π΅Π»Ρ ΡΠ°Π±ΠΎΡΡ: ΡΠ°Π·ΡΠ°Π±ΠΎΡΠΊΠ° ΠΌΠ΅ΡΠΎΠ΄ΠΈΠΊΠΈ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΡ ΡΠΎΠ΄ΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΡΠΈΠΌΠ΅ΡΠ΅ΠΉ ΠΈ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠ³ΠΎ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΡ ΠΏΠ°ΠΏΠ°Π²Π΅ΡΠΈΠ½Π° Π³ΠΈΠ΄ΡΠΎΡ
Π»ΠΎΡΠΈΠ΄Π° Π² ΡΠ°ΡΡΠ²ΠΎΡΠ΅ Π΄Π»Ρ ΠΈΠ½ΡΠ΅ΠΊΡΠΈΠΉ, ΡΠ°Π±Π»Π΅ΡΠΊΠ°Ρ
ΠΈ ΡΡΠΏΠΏΠΎΠ·ΠΈΡΠΎΡΠΈΡΡ
ΡΠ΅ΠΊΡΠ°Π»ΡΠ½ΡΡ
ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΠΠΠ₯.ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ: ΠΎΠ±ΡΠ΅ΠΊΡΠ°ΠΌΠΈ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ ΡΠ²Π»ΡΠ»ΠΈΡΡ ΠΎΠ±ΡΠ°Π·ΡΡ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ² ΠΏΠ°ΠΏΠ°Π²Π΅ΡΠΈΠ½Π° ΠΎΡΠ΅ΡΠ΅ΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΡΠΎΠΈΠ·Π²ΠΎΠ΄ΠΈΡΠ΅Π»Π΅ΠΉ: Β«ΠΠ°ΠΏΠ°Π²Π΅ΡΠΈΠ½, ΡΠ°ΡΡΠ²ΠΎΡ Π΄Π»Ρ ΠΈΠ½ΡΠ΅ΠΊΡΠΈΠΉ 20Β ΠΌΠ³/ΠΌΠ»Β», Β«ΠΠ°ΠΏΠ°Π²Π΅ΡΠΈΠ½, ΡΡΠΏΠΏΠΎΠ·ΠΈΡΠΎΡΠΈΠΈ ΡΠ΅ΠΊΡΠ°Π»ΡΠ½ΡΠ΅, 20Β ΠΌΠ³Β», Β«ΠΠ°ΠΏΠ°Π²Π΅ΡΠΈΠ½, ΡΠ°Π±Π»Π΅ΡΠΊΠΈ 40Β ΠΌΠ³Β». ΠΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠ΅ ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΠΠΠ₯ Π½Π° ΠΆΠΈΠ΄ΠΊΠΎΡΡΠ½ΠΎΠΌ Ρ
ΡΠΎΠΌΠ°ΡΠΎΠ³ΡΠ°ΡΠ΅ Agilent 1260 Infinity II DAD System , ΡΠ΅Π³ΠΈΡΡΡΠ°ΡΠΈΡ Π£Π€-ΡΠΏΠ΅ΠΊΡΡΠΎΠ² β Π½Π° ΡΠΏΠ΅ΠΊΡΡΠΎΡΠΎΡΠΎΠΌΠ΅ΡΡΠ΅ Agilent 8453Π UV-Vis System. ΠΡΠ΅Π½ΠΊΡ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΡ ΡΠΎΠ΄ΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΡΠΈΠΌΠ΅ΡΠ΅ΠΉ ΠΈ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠ΅ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΠ΅ Π΄Π΅ΠΉΡΡΠ²ΡΡΡΠΈΡ
Π²Π΅ΡΠ΅ΡΡΠ² ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ ΠΎΠ΄Π½ΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½ΠΎ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΠΠΠ₯ Π½Π° ΠΎΠ±ΡΠ°ΡΠ΅Π½Π½ΠΎ-ΡΠ°Π·ΠΎΠ²ΠΎΠΉ ΠΊΠΎΠ»ΠΎΠ½ΠΊΠ΅ Kromasil 100-5-C18, ΡΠ°Π·ΠΌΠ΅Ρ ΠΊΠΎΠ»ΠΎΠ½ΠΊΠΈ 250Γ4,6Β ΠΌΠΌ, ΡΠ°Π·ΠΌΠ΅Ρ ΡΠ°ΡΡΠΈΡ 5Β ΠΌΠΊΠΌ, Π² ΡΡΠ»ΠΎΠ²ΠΈΡΡ
Π³ΡΠ°Π΄ΠΈΠ΅Π½ΡΠ½ΠΎΠ³ΠΎ ΡΠ»ΡΠΈΡΠΎΠ²Π°Π½ΠΈΡ Ρ Π΄Π΅ΡΠ΅ΠΊΡΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΠΏΡΠΈ 238Β Π½ΠΌ. Π ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅ ΡΡΠ°Π½Π΄Π°ΡΡΠ½ΡΡ
ΠΎΠ±ΡΠ°Π·ΡΠΎΠ² ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π»ΠΈ ΠΏΠ°ΠΏΠ°Π²Π΅ΡΠΈΠ½Π° Π³ΠΈΠ΄ΡΠΎΡ
Π»ΠΎΡΠΈΠ΄ ΠΊΠ²Π°Π»ΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ USP RS Ρ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΠ΅ΠΌ Π΄Π΅ΠΉΡΡΠ²ΡΡΡΠ΅Π³ΠΎ Π²Π΅ΡΠ΅ΡΡΠ²Π° 99,9% ΠΈ Π½ΠΎΡΠΊΠ°ΠΏΠΈΠ½ ΠΊΠ²Π°Π»ΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ EP CRS.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ: ΠΏΠΎΠΊΠ°Π·Π°Π½Π° Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎΡΡΡ ΠΎΠ΄Π½ΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½ΠΎΠ³ΠΎ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΡ ΡΠΎΠ΄ΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΡΠΈΠΌΠ΅ΡΠ΅ΠΉ ΠΈ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠ³ΠΎ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΡ Π΄Π΅ΠΉΡΡΠ²ΡΡΡΠΈΡ
Π²Π΅ΡΠ΅ΡΡΠ² Π² ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ°Ρ
ΠΏΠ°ΠΏΠ°Π²Π΅ΡΠΈΠ½Π° ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΠΠΠ₯.ΠΡΠ²ΠΎΠ΄Ρ: ΠΏΡΠ΅Π΄Π»ΠΎΠΆΠ΅Π½Π° ΠΌΠ΅ΡΠΎΠ΄ΠΈΠΊΠ° ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΡ Π΄Π΅ΠΉΡΡΠ²ΡΡΡΠΈΡ
Π²Π΅ΡΠ΅ΡΡΠ² Π² ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ°Ρ
ΠΏΠ°ΠΏΠ°Π²Π΅ΡΠΈΠ½Π° ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΠΠΠ₯, ΡΠ΄ΠΎΠ²Π»Π΅ΡΠ²ΠΎΡΡΡΡΠ°Ρ ΠΏΡΠΈΠ½ΡΠΈΠΏΡ ΡΠΊΠ²ΠΎΠ·Π½ΠΎΠΉ ΡΡΠ°Π½Π΄Π°ΡΡΠΈΠ·Π°ΡΠΈΠΈ, ΠΊΠΎΡΠΎΡΠ°Ρ ΠΌΠΎΠΆΠ΅Ρ Π±ΡΡΡ ΡΠ΅ΠΊΠΎΠΌΠ΅Π½Π΄ΠΎΠ²Π°Π½Π° Π΄Π»Ρ Π²ΠΊΠ»ΡΡΠ΅Π½ΠΈΡ Π² ΠΏΡΠΎΠ΅ΠΊΡ ΡΠ°ΡΠΌΠ°ΠΊΠΎΠΏΠ΅ΠΉΠ½ΡΡ
ΡΡΠ°ΡΠ΅ΠΉ Π½Π° ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΡ ΠΏΠ°ΠΏΠ°Π²Π΅ΡΠΈΠ½Π°
Π‘ΡΠ°Π²Π½ΠΈΡΠ΅Π»ΡΠ½Π°Ρ ΠΎΡΠ΅Π½ΠΊΠ° ΡΡΠ΅Π±ΠΎΠ²Π°Π½ΠΈΠΉ ΠΊ ΠΊΠ°ΡΠ΅ΡΡΠ²Ρ Π»Π΅ΠΊΠ°ΡΡΡΠ²Π΅Π½Π½ΡΡ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ², ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΡ Π΄ΠΈΠΎΡΠΌΠΈΠ½
Currently, there is an increase in pre- and post-approval testing of medicinal products containing diosmin and hence a need to unify approaches to standardisation of this group of pharmaceuticals. Moreover, the State Pharmacopoeia of the Russian Federation lacks a monograph for these products.The aim of the study was to determine an approach to standardisation of medicinal products containing diosmin.Materials and methods: the study analysed scientific publications, as well as monographs of leading foreign pharmacopoeias. Experimental work was carried out using samples of diosmin-containing pharmaceuticals in the form of 500 and 1000 mg film-coated tablets produced by Russian and foreign manufacturers. The study involved high performance liquid chromatography with UV detection using an Agilent 1260 Infinity II liquid chromatography system with a diode array detector. The following reference standards were used: a diosmin RS, USP grade; a hesperidin CRS, Ph. Eur. Grade; and a diosmin CRS for testing chromatography system suitability for identification of impurities A, B, C, D, E, and F, Ph. Eur. grade.Results: the authors reviewed quality requirements for pharmaceutical products containing diosmin and analysed experimental data obtained during pre- and post-approval testing of Russian and foreign medicines. The comparison of regulatory documents for registered diosmin-containing medicinal products showed a difference in approaches to assessing the contents of related substances and active pharmaceutical ingredients. Having analysed the literature, experimental data and regulatory requirements for standardisation of diosmin-containing pharmaceuticals, the authors recommended an approach to standardisation. According to the approach, concomitant flavonoids (hesperidin, isorchoifolin, linarin, and diosmetin) contributing to the pharmacological activity of a medicinal product are specified as part of Assay, and process-related by-products (impurities A and D) are specified and evaluated as part of Related substances tests.Conclusion: the authors propose to evaluate the contents of concomitant flavonoids (hesperidin, isorchoifolin, linarin, diosmetin) under Assay and to specify impurities A and D, as well as single unidentified impurities and total amount of impurities under Related substances.Π Π½Π°ΡΡΠΎΡΡΠ΅Π΅ Π²ΡΠ΅ΠΌΡ ΡΡΡΠ΅ΡΡΠ²ΡΠ΅Ρ ΠΏΠΎΡΡΠ΅Π±Π½ΠΎΡΡΡ Π² ΡΠ½ΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ ΠΏΠΎΠ΄Ρ
ΠΎΠ΄ΠΎΠ² ΠΊ ΡΡΠ°Π½Π΄Π°ΡΡΠΈΠ·Π°ΡΠΈΠΈ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ² Π΄ΠΈΠΎΡΠΌΠΈΠ½Π° Π² ΡΠ²ΡΠ·ΠΈ Ρ ΡΠ²Π΅Π»ΠΈΡΠ΅Π½ΠΈΠ΅ΠΌ ΠΎΠ±ΡΠ΅ΠΌΠ° ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½ΠΈΡ ΡΠ΅Π³ΠΈΡΡΡΠ°ΡΠΈΠΎΠ½Π½ΠΎΠΉ ΠΈ ΠΏΠΎΡΡΡΠ΅Π³ΠΈΡΡΡΠ°ΡΠΈΠΎΠ½Π½ΠΎΠΉ ΡΠΊΡΠΏΠ΅ΡΡΠΈΠ·Ρ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ² ΡΡΠΎΠΉ Π³ΡΡΠΏΠΏΡ, Π° ΡΠ°ΠΊΠΆΠ΅ Ρ Π½Π΅ΠΎΠ±Ρ
ΠΎΠ΄ΠΈΠΌΠΎΡΡΡΡ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠΊΠΈ ΡΠ°ΡΠΌΠ°ΠΊΠΎΠΏΠ΅ΠΉΠ½ΠΎΠΉ ΡΡΠ°ΡΡΠΈ Π½Π° ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΡ Π΄ΠΈΠΎΡΠΌΠΈΠ½Π° Π΄Π»Ρ ΠΠΎΡΡΠ΄Π°ΡΡΡΠ²Π΅Π½Π½ΠΎΠΉ ΡΠ°ΡΠΌΠ°ΠΊΠΎΠΏΠ΅ΠΈ Π ΠΎΡΡΠΈΠΉΡΠΊΠΎΠΉ Π€Π΅Π΄Π΅ΡΠ°ΡΠΈΠΈ.Π¦Π΅Π»Ρ ΡΠ°Π±ΠΎΡΡ: ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΠ΅ ΠΏΠΎΠ΄Ρ
ΠΎΠ΄Π° ΠΊ ΡΡΠ°Π½Π΄Π°ΡΡΠΈΠ·Π°ΡΠΈΠΈ Π»Π΅ΠΊΠ°ΡΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ², ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΡ
Π΄ΠΈΠΎΡΠΌΠΈΠ½.ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ: ΠΎΠ±ΡΠ΅ΠΊΡΠ°ΠΌΠΈ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ ΡΠ»ΡΠΆΠΈΠ»ΠΈ Π΄Π°Π½Π½ΡΠ΅ Π½Π°ΡΡΠ½ΡΡ
ΠΏΡΠ±Π»ΠΈΠΊΠ°ΡΠΈΠΉ, Π° ΡΠ°ΠΊΠΆΠ΅ ΡΠ°ΡΡΠ½ΡΠ΅ ΠΌΠΎΠ½ΠΎΠ³ΡΠ°ΡΠΈΠΈ Π²Π΅Π΄ΡΡΠΈΡ
Π·Π°ΡΡΠ±Π΅ΠΆΠ½ΡΡ
ΡΠ°ΡΠΌΠ°ΠΊΠΎΠΏΠ΅ΠΉ. ΠΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°Π»ΡΠ½Π°Ρ ΡΠ°Π±ΠΎΡΠ° ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»Π°ΡΡ Π½Π° ΠΎΠ±ΡΠ°Π·ΡΠ°Ρ
ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ² Π΄ΠΈΠΎΡΠΌΠΈΠ½Π° Π² ΡΠΎΡΠΌΠ΅ ΡΠ°Π±Π»Π΅ΡΠΎΠΊ, ΠΏΠΎΠΊΡΡΡΡΡ
ΠΏΠ»Π΅Π½ΠΎΡΠ½ΠΎΠΉ ΠΎΠ±ΠΎΠ»ΠΎΡΠΊΠΎΠΉ, Π² Π΄ΠΎΠ·ΠΈΡΠΎΠ²ΠΊΠ΅ 500 ΠΈ 1000 ΠΌΠ³, ΠΎΡΠ΅ΡΠ΅ΡΡΠ²Π΅Π½Π½ΡΡ
ΠΈ Π·Π°ΡΡΠ±Π΅ΠΆΠ½ΡΡ
ΠΏΡΠΎΠΈΠ·Π²ΠΎΠ΄ΠΈΡΠ΅Π»Π΅ΠΉ. ΠΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠ΅ ΠΎΡΡΡΠ΅ΡΡΠ²Π»ΡΠ»ΠΈ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ Π²ΡΡΠΎΠΊΠΎΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΠΉ ΠΆΠΈΠ΄ΠΊΠΎΡΡΠ½ΠΎΠΉ Ρ
ΡΠΎΠΌΠ°ΡΠΎΠ³ΡΠ°ΡΠΈΠΈ Ρ Π£Π€-Π΄Π΅ΡΠ΅ΠΊΡΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ Π½Π° ΠΆΠΈΠ΄ΠΊΠΎΡΡΠ½ΠΎΠΌ Ρ
ΡΠΎΠΌΠ°ΡΠΎΠ³ΡΠ°ΡΠ΅ Agilent 1260 Infinity II DAD System. Π ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅ ΡΡΠ°Π½Π΄Π°ΡΡΠ½ΡΡ
ΠΎΠ±ΡΠ°Π·ΡΠΎΠ² ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π»ΠΈ Π΄ΠΈΠΎΡΠΌΠΈΠ½ ΠΊΠ²Π°Π»ΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ USP RS, Π³Π΅ΡΠΏΠ΅ΡΠΈΠ΄ΠΈΠ½ ΠΊΠ²Π°Π»ΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ EP CRS ΠΈ Π΄ΠΈΠΎΡΠΌΠΈΠ½ Π΄Π»Ρ ΠΏΡΠΎΠ²Π΅ΡΠΊΠΈ ΠΏΡΠΈΠ³ΠΎΠ΄Π½ΠΎΡΡΠΈ Ρ
ΡΠΎΠΌΠ°ΡΠΎΠ³ΡΠ°ΡΠΈΡΠ΅ΡΠΊΠΎΠΉ ΡΠΈΡΡΠ΅ΠΌΡ ΠΊΠ²Π°Π»ΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ EP CRS Π΄Π»Ρ ΠΈΠ΄Π΅Π½ΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ ΠΏΡΠΈΠΌΠ΅ΡΠ΅ΠΉ A, B, C, D, E ΠΈ F.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ: ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½ Π°Π½Π°Π»ΠΈΡΠΈΡΠ΅ΡΠΊΠΈΠΉ ΠΎΠ±Π·ΠΎΡ ΡΡΠ΅Π±ΠΎΠ²Π°Π½ΠΈΠΉ ΠΊ ΠΊΠ°ΡΠ΅ΡΡΠ²Ρ Π»Π΅ΠΊΠ°ΡΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ², ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΡ
Π΄ΠΈΠΎΡΠΌΠΈΠ½, Ρ Π°Π½Π°Π»ΠΈΠ·ΠΎΠΌ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°Π»ΡΠ½ΡΡ
Π΄Π°Π½Π½ΡΡ
, ΠΏΠΎΠ»ΡΡΠ΅Π½Π½ΡΡ
Π² Ρ
ΠΎΠ΄Π΅ ΡΠ΅Π³ΠΈΡΡΡΠ°ΡΠΈΠΎΠ½Π½ΠΎΠΉ ΠΈ ΠΏΠΎΡΡΡΠ΅Π³ΠΈΡΡΡΠ°ΡΠΈΠΎΠ½Π½ΠΎΠΉ ΡΠΊΡΠΏΠ΅ΡΡΠΈΠ·Ρ ΡΠΎΡΡΠΈΠΉΡΠΊΠΈΡ
ΠΈ Π·Π°ΡΡΠ±Π΅ΠΆΠ½ΡΡ
Π»Π΅ΠΊΠ°ΡΡΡΠ²Π΅Π½Π½ΡΡ
ΡΡΠ΅Π΄ΡΡΠ². Π‘ΡΠ°Π²Π½ΠΈΡΠ΅Π»ΡΠ½ΡΠΉ Π°Π½Π°Π»ΠΈΠ· Π½ΠΎΡΠΌΠ°ΡΠΈΠ²Π½ΠΎΠΉ Π΄ΠΎΠΊΡΠΌΠ΅Π½ΡΠ°ΡΠΈΠΈ Π·Π°ΡΠ΅Π³ΠΈΡΡΡΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
Π»Π΅ΠΊΠ°ΡΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ², ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΡ
Π΄ΠΈΠΎΡΠΌΠΈΠ½, ΠΏΠΎΠΊΠ°Π·Π°Π» ΡΠ°Π·Π»ΠΈΡΠΈΠ΅ Π² ΠΏΠΎΠ΄Ρ
ΠΎΠ΄Π°Ρ
ΠΊ ΠΎΡΠ΅Π½ΠΊΠ΅ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΡ ΡΠΎΠ΄ΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΡΠΈΠΌΠ΅ΡΠ΅ΠΉ ΠΈ Π΄Π΅ΠΉΡΡΠ²ΡΡΡΠΈΡ
Π²Π΅ΡΠ΅ΡΡΠ². ΠΠ½Π°Π»ΠΈΠ· Π΄Π°Π½Π½ΡΡ
Π»ΠΈΡΠ΅ΡΠ°ΡΡΡΡ, ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°Π»ΡΠ½ΡΡ
Π΄Π°Π½Π½ΡΡ
ΠΈ ΡΡΠ΅Π±ΠΎΠ²Π°Π½ΠΈΠΉ Π½ΠΎΡΠΌΠ°ΡΠΈΠ²Π½ΡΡ
Π΄ΠΎΠΊΡΠΌΠ΅Π½ΡΠΎΠ² Π² ΠΎΠ±Π»Π°ΡΡΠΈ ΡΡΠ°Π½Π΄Π°ΡΡΠΈΠ·Π°ΡΠΈΠΈ, ΠΏΡΠ΅Π΄ΡΡΠ²Π»ΡΠ΅ΠΌΡΡ
ΠΊ ΡΡΠ°Π½Π΄Π°ΡΡΠΈΠ·Π°ΡΠΈΠΈ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ², ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΡ
Π΄ΠΈΠΎΡΠΌΠΈΠ½, ΠΏΠΎΠ·Π²ΠΎΠ»ΡΠ΅Ρ ΡΠ΅ΠΊΠΎΠΌΠ΅Π½Π΄ΠΎΠ²Π°ΡΡ ΠΏΠΎΠ΄Ρ
ΠΎΠ΄, ΡΠΎΠ³Π»Π°ΡΠ½ΠΎ ΠΊΠΎΡΠΎΡΠΎΠΌΡ ΡΠΎΠΏΡΡΡΡΠ²ΡΡΡΠΈΠ΅ ΡΠ»Π°Π²ΠΎΠ½ΠΎΠΈΠ΄Ρ (Π³Π΅ΡΠΏΠ΅ΡΠΈΠ΄ΠΈΠ½, ΠΈΠ·ΠΎΡΡ
ΠΎΠΉΡΠΎΠ»ΠΈΠ½, Π»ΠΈΠ½Π°ΡΠΈΠ½, Π΄ΠΈΠΎΡΠΌΠ΅ΡΠΈΠ½), ΠΎΠ±Π΅ΡΠΏΠ΅ΡΠΈΠ²Π°ΡΡΠΈΠ΅ Π²ΠΊΠ»Π°Π΄ Π² ΡΠ°ΡΠΌΠ°ΠΊΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠ΅ Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ°, Π½ΠΎΡΠΌΠΈΡΡΡΡΡΡ Π² ΡΠΎΡΡΠ°Π²Π΅ ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»Ρ Β«ΠΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠ΅ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΠ΅Β». ΠΡΠΈΠΌΠ΅ΡΠΈ, ΠΎΡΠ½ΠΎΡΡΡΠΈΠ΅ΡΡ ΠΊ ΠΏΠΎΠ±ΠΎΡΠ½ΡΠΌ ΠΏΡΠΎΠ΄ΡΠΊΡΠ°ΠΌ ΠΏΡΠΈ ΠΏΡΠΎΠΈΠ·Π²ΠΎΠ΄ΡΡΠ²Π΅ Π΄ΠΈΠΎΡΠΌΠΈΠ½Π° (ΠΏΡΠΈΠΌΠ΅ΡΠΈ Π ΠΈ D), Π½ΠΎΡΠΌΠΈΡΡΡΡΡΡ ΠΈ ΠΎΡΠ΅Π½ΠΈΠ²Π°ΡΡΡΡ ΠΏΡΠΈ ΠΈΡΠΏΡΡΠ°Π½ΠΈΡΡ
ΠΏΠΎ ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»Ρ Β«Π ΠΎΠ΄ΡΡΠ²Π΅Π½Π½ΡΠ΅ ΠΏΡΠΈΠΌΠ΅ΡΠΈΒ».ΠΡΠ²ΠΎΠ΄Ρ: ΠΏΡΠ΅Π΄Π»ΠΎΠΆΠ΅Π½ΠΎ ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΡΡ ΠΎΡΠ΅Π½ΠΊΡ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΡ ΡΠΎΠΏΡΡΡΡΠ²ΡΡΡΠΈΡ
ΡΠ»Π°Π²ΠΎΠ½ΠΎΠΈΠ΄ΠΎΠ² (Π³Π΅ΡΠΏΠ΅ΡΠΈΠ΄ΠΈΠ½Π°, ΠΈΠ·ΠΎΡΡ
ΠΎΠΉΡΠΎΠ»ΠΈΠ½Π°, Π»ΠΈΠ½Π°ΡΠΈΠ½Π°, Π΄ΠΈΠΎΡΠΌΠ΅ΡΠΈΠ½Π°) Π² ΡΠΎΡΡΠ°Π²Π΅ ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»Ρ Β«ΠΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠ΅ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΠ΅Β», ΠΏΡΠΈΠΌΠ΅ΡΠΈ Π ΠΈ D, Π° ΡΠ°ΠΊΠΆΠ΅ Π΅Π΄ΠΈΠ½ΠΈΡΠ½ΡΠ΅ Π½Π΅ΠΈΠ΄Π΅Π½ΡΠΈΡΠΈΡΠΈΡΠΎΠ²Π°Π½Π½ΡΠ΅ ΠΏΡΠΈΠΌΠ΅ΡΠΈ ΠΈ ΡΡΠΌΠΌΡ ΠΏΡΠΈΠΌΠ΅ΡΠ΅ΠΉ Π½ΠΎΡΠΌΠΈΡΠΎΠ²Π°ΡΡ Π² ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»Π΅ Β«Π ΠΎΠ΄ΡΡΠ²Π΅Π½Π½ΡΠ΅ ΠΏΡΠΈΠΌΠ΅ΡΠΈΒ»
Evaluation of the diagnostic efficacy of a reagent kit for <i>in vitro</i> diagnosis of West Nile fever using reverse transcription polymerase chain reaction with fluorescent probe-based detection
West Nile fever is a vector-borne zoonotic arbovirus infection with natural foci. Its clinical course is similar to that of acute febrile syndrome, and severe cases may result in neuroinvasive disease. Several genetic lineages (1, 2, and 4) of the West Nile virus (WNV) with different pathogenicity for humans are circulating in the Russian Federation. Therefore, it is an urgent task to develop a diagnostic reagent kit for differentiating between WNV genetic lineages and to implement the kit in clinical laboratory practice.The aim of the study was to conduct technical and clinical tests and evaluate the quality, efficacy, and safety of the Ampligen-WNV-genotype-1/2/4 diagnostic reagent kit for detecting WNV RNA and differentiating between WNV genetic lineages 1, 2, and 4 by reverse transcription polymerase chain reaction (RT-PCR) with fluorescent probe-based detection.Materials and methods. The authors determined the diagnostic sensitivity and specificity of the Ampligen-WNV-genotype-1/2/4 reagent kit (Volgograd Research Institute for Plague Control, Russia) by real-time RT-PCR with 216 clinical samples and 204 biological samples. Sanger sequencing was used as a reference method. Statistical analysis of clinical test results was carried out in accordance with the Russian national standard for clinical laboratory tests (GOST R 53022.3-2008).Results. When tested with the Ampligen-WNV-genotype-1/2/4 reagent kit, real-time RT-PCR demonstrated the analytical sensitivity of 1Γ104 Β GEq/mL for the detection of WNV cDNA of genetic lineages 1, 2, and 4. The assessment of its analytical specificity showed no positive results for cDNA samples of heterologous viruses at a concentration of 1Γ106 Β GEq/mL. The diagnostic sensitivity with the reagent kit was at least 98.5%, and the diagnostic specificity was at least 99%, with 90% confidence levels for both parameters.Conclusions. The Ampligen-WNV-genotype-1/2/4 reagent kit can be recommended for use in clinical laboratory diagnostics to detect WNV RNA and differentiate between WNV genetic lineages 1, 2, and 4
Improvement of Methods of Standardisation of Medicinal Products Made from Veratrum Lobelianum Rhizomes with Roots
Abstract. Identification of hellebore (Veratrum Lobelianum Bernh.) herbal substance, as well as hellebore-based herbal preparation and herbal medicinal product by the same group of biologically active substances using the same test method is in line with the so-called βconsistent standardisationβ principle.The aim of the study was to develop a harmonised approach to identification of steroidal alkaloids in hellebore products (hellebore water, hellebore tincture) and hellebore herbal substance (hellebore rhizomes with roots).Materials and methods: samples of hellebore water, hellebore tincture, and hellebore rhizomes with roots were analysed by high-performance thin-layer chromatography (HPTLC) using an HPTLC plate.Results: the authors developed a harmonised identification procedure for products made from hellebore rhizomes with roots (herbal substance, herbal preparation, and herbal medicinal product) based on HPTLC detection of steroidal alkaloids. The results of the study will be used to prepare amendments to the Identification part of monograph FS.2.5.0104.18 βHellebore rhizomes with rootsβ. The developed test procedure is proposed for inclusion into draft monographs βHellebore rhizomes with roots, tinctureβ and βHellebore rhizomes with roots, tincture, solution for external useβ.Conclusions: the developed test procedure can be used as an identification test for a range of products from the hellebore herbal substance to hellebore-based herbal medicinal products, which is based on the detection of the same group of biologically active substances
Development of a Comprehensive HPLC Method for Determination of Product-related Impurities and Assay of Active Ingredients in Papaverine Hydrochloride Products
Abstract. Papaverine hydrochloride products are used as anticonvulsants in routine medical practice. Most of the approved product specification files include thin-layer chromatography for assessment of product-related impurities and UV spectrophotometry for determination of active pharmaceutical ingredients. An HPLC assay is not used for determination of papaverine hydrochloride in drug dosage forms.The aim of the study was to develop an HPLC test method for determination of product-related impurities and for quantification of papaverine hydrochloride in solutions for injection, tablets, and rectal suppositories.Materials and methods: samples of the following Russian-made papaverine products were used in the study: Papaverine, solution for injection, 20Β mg/mL; Papaverine, rectal suppositories, 20Β mg; Papaverine, tablets, 40Β mg. The Agilent 1260 Infinity II DAD System was used for the HPLC assay, and the Agilent 8453Π UV-Vis System was used for recording UV spectra. The determination of product-related impurities and the assay of active ingredients were performed simultaneously by HPLC using a reversed-phase column Kromasil 100-5-C18, 250Γ4.6Β mm, 5 ΞΌm, the gradient elution mode, and detection atΒ Β Β Β 238 nm. Papaverine Hydrochloride USP RS, 99% purity, and Noscapine EP CRS were used as reference standards.Results: the study demonstrated that determination of product-related impurities and assay of active ingredients in papaverine products can be performed simultaneously using HPLC.Conclusions: the authors proposed an HPLC test method for determination of active ingredients in papaverine products, which is aligned with the βconsistent standardisationβ principle and can be recommended for inclusion into draft monographs for papaverine products