States Pharmacopeia Research Articles

Certification of a pharmacopoeial reference standard for potency testing of recombinant interferon α-2b

SCIENTIFIC RELEVANCE. Potency testing of recombinant interferons requires a reference standard. The availability of International Standards (ISs) that are commonly used to assess the quality of recombinant interferons is currently limited. Therefore, the quality of interferons should be assessed using pharmacopoeial reference standards (RSs) certified using ISs (if available).AIM. This study aimed to certify a pharmacopoeial RS for potency testing of recombinant interferon α-2b.MATERIALS AND METHODS. The potency determination involved comparing the inhibition of the virus-induced cytopathic effect observed in cell culture with the candidate RS and the WHO International Standard for Human rDNA-derived interferon α-2b. The study used A-549 and MDBK cell lines with encephalomyocarditis (EMC) and vesicular stomatitis (VSV) viruses. The authors followed the requirements of the State Pharmacopoeia of the Russian Federation (Cell-Culture Bioassays for Interferon Products, OFS.1.7.2.0002.15). The results were recorded using instrumental and visual methods. The calculations used mathematical statistics. To factor in the influence of intermediate precision, the authors applied Student’s t-test.RESULTS. The authors developed a certification programme and procedure and certified the pharmacopoeial RS for recombinant human interferon α-2b (rhIFN α-2b) to identify the corresponding medicinal products (by virus neutralisation) and test their potency (by antiviral activity in cell culture). Upon receipt, the candidate RS was verified for compliance with regulatory specifications. According to the test results, the potency of the candidate pharmacopoeial RS was 4.47×108 IU/mL, and the expanded uncertainty was 8.12×107 IU/mL, with a coverage factor of k=2 and a confidence level of 95%. The pharmacopoeial RS for rhIFN α-2b was considered to have the same shelf-life period as the corresponding medicinal products if stored at a temperature not higher than –40 ºC. When thawed, the pharmacopoeial RS for rhIFN α-2b can be stored at a temperature of 2–8 ºC for up to 1 month.CONCLUSIONS. Upon certification for potency testing, the pharmacopoeial RS for rhIFN α-2b has been included in the Register of Reference Standards of the Russian Pharmacopoeia as Recombinant Human Interferon α-2b (Potency) FSO.3.2.00455. The introduction of this pharmacopoeial RS will help to conduct the identification and potency testing of all Russian interferon α-2b products at an appropriate scientific and methodological level. As a result, Russian pharmaceutical manufacturers and quality control agencies will no longer depend on imported ISs.

Comparison of In vitro Dissolution Tests of five brands of Diclofenac Sodium delayed release tablets in Moroccan market

This study aims to evaluate the similarity of in-vitro dissolution profiles, of commercially available generics of diclofenac sodium delayed-release tablets, of 25mg strength, with the innovator brand in Moroccan Market. Dissolution profiles for five brands were determined using the paddle method from the United States Pharmacopoeia. For the aptitude verification of the method, in our experimental conditions, linearity and accuracy tests were performed. The rate of dissolution was determined by Ultra-Violet Spectroscopy at 276nm. The assessment between the generics and the brand medicine was carried out under the similarity factor approach. We also presented an overview of the dissolution principles of tablet dosage form, and the pharmacopoeia dissolution test recommendations in our case. To compliance with US pharmacopoeia specifications, the rate of dissolution should be not more than 10% after 2hours in the acid stage, and not less than 75% at 45 minutes in the buffer stage. Furthermore, two dissolution profiles are considered similar when the f2 value is ≥50. According to routine quality control, evaluating the rate dissolution at the final point, all the generic medicines marketed in Morocco, meet the specification of the dissolution testing. However, using the comparison of dissolution profiles, only generic-2 is interchangeable with the innovator brand.

Development and validation of an analytical procedure for the determination of residual protein A in active substances of therapeutic monoclonal antibodies

SCIENTIFIC RELEVANCE. An important quality-control issue for therapeutic monoclonal antibodies (mAbs) is the determination of residual protein A leaching from the carrier during the purification of mAbs by affinity chromatography. However, unrelated compounds (immunoglobulins) present in the sample can complicate the immunoenzymatic detection of protein A (matrix effect), potentially leading to false-negative test results. To increase the efficiency of enzyme-linked immunosorbent assay (ELISA), it is necessary to develop a sample preparation step that can irreversibly break the bond in the protein A–mAb complex.AIM. This study aimed to develop and validate an analytical procedure for the determination of residual protein A in active substances of therapeutic mAbs by ELISA with a test kit comprising in-house reagents.MATERIALS AND METHODS. Recombinant protein A was used as an antigen. Polyclonal antibodies to protein A were produced by immunising chickens, selecting immunised eggs, and isolating antibodies from these eggs. Protein A-specific antibodies were purified by affinity chromatography. Residual protein A was determined using sandwich ELISA with preliminary sample preparation. The analytical procedure was validated in accordance with the requirements of the State Pharmacopoeia of the Russian Federation (Validation of Analytical Procedures, OFS.1.1.0012).RESULTS. The authors obtained, isolated, and purified chicken IgY antibodies to recombinant protein A. The authors selected sample preparation conditions for the determination of residual protein A by ELISA and optimum compositions of buffer solutions, including the composition of a denaturing buffer to disrupt the protein A–mAb complex. The developed analytical procedure was validated. According to the measurements of protein A, the accuracy of the analytical procedure ranged within 83–108% of the nominal value, the interlaboratory precision ranged within 96–116%, and the repeatability was up to 13%. The lower limit of quantitation was 10 ng/mL with a minimum required dilution of 1:10. The analytical range extended from 10 to 40 ng/mL. The analytical procedure showed results comparable to those obtained with a similar test kit from an international manufacturer.CONCLUSIONS. The developed analytical procedure for the determination of residual protein A by ELISA with a test kit comprising in-house reagents can minimise the matrix effect in therapeutic mAbs. This analytical procedure will alleviate import substitution and reduce quality control costs for Russian immunobiologicals.

A COMPARATIVE STUDY OF THE COMPONENT COMPOSITION OF THE HERBS OF SOME SPECIES OF THE GENUS ACHILLEA L.

The article presents the results of a comparative study of the component composition of herbs of some species of the genus Achillea L. growing in the Samara region - common yarrow (Achillea millefolium L.), cartilaginous yarrow (Achillea cartilaginea (Ledeb. ex Rchb.) and noble yarrow (Achillea nobilis (L.). Spectrophotometry and high performance liquid chromatography were used as methods of investigation.
 As a result of comparative spectrophotometric analysis of water-alcoholic extracts of common yarrow herbs, yarrow cartilaginous herbs and noble yarrow herbs revealed that the absorption curve of their UV spectra is due mainly to hydroxycinnamic alcohols, in particular chlorogenic and caffeic acids (290 sh. and 330 nm). When aluminum (III) chloride solution was added to the water-alcoholic extracts of yarrow herb from the studied species, a bathochromic shift of the long-wave band at 400 nm was observed due to the flavonoids.
 The content of total flavonoids in the plant raw material is: common yarrow - 0.68±0.01%, cartilaginous yarrow - 0.65±0.02%, noble yarrow - 0.69±0.02%.
 Chlorogenic acid, cynaroside, cosmosiin, apigenin and luteolin were identified in water-alcoholic extracts from the above described species using HPLC method. In addition, the presence of caffeic acid was detected in the water-alcoholic extracts of noble yarrow herbs.
 The content of cosmosiin in the plant raw material is: common yarrow - 0.61±0.01%, cartilaginous yarrow - 0.56±0.02%, noble yarrow - 0.62±0.02%.
 The results of the study can be used in the development of regulatory documentation for medicinal plant material "Yarrow herb" for inclusion in the State Pharmacopoeia of the Russian Federation.

CRITICAL ANALYSIS OF POWDER FLOW BEHAVIOUR OF DIRECTLY COMPRESSIBLE COPROCESSED EXCIPIENTS

The aim of this pre-formulation study was to adopt simple linear regression modelling and correlation statistics to understand the associations between pharmacopoeial powder test methods using datasets generated from five commercial brands of directly compressible excipients with a specific focus to inferential implications in formulation design. Powder characterization was conducted using protocols defined in Chapter <1174> and <616> of the United States Pharmacopoeia (USP41-NF36). The study adopted a linear regression modelling analytics and correlation statistics using the fitting algorithm of OriginPro® (OriginPro, Version 2021b, OriginLab Corporation, Northampton, MA, USA). In the results, the modulus of Pearson’s product moment correlation coefficient was used to measure the strength of the linear association between test variables and a correlation matrix generated. Strong positive correlation modulus of Hausner’s Ratio (HR) with Carr’s index (r=+0.999) and static angle of repose (r=+0.932) were evident. Bulk density strongly correlates with tap density in the positive direction (r=+0.911). Tap density also shows a slight negative correlation with HR (r=-0.230), Carr’s index (r=-0.228), and static angle of repose (r==-0.421), while Carr’s index strongly correlated with static angle of repose (r=+0.933). In conclusion, modelling bivariate powder flow datasets has provided a powerful but simplistic statistical relationship for characterizing the modulus of association between HR, Carr’s index, and static angle of repose of the model excipients useful in preformulation design of pharmaceutical formulations.

Optimization of the thin layer chromatography method conditions for the detection of related dioxidine impurities

When developing methods for analysing active pharmaceutical substances (API), it is necessary to take into account modern requirements for the content of impurities in them. At the same time, for some drugs that have been on the pharmaceutical market for a long time, analysis methods are still used that are unable to provide the necessary sensitivity and specificity. One of these API is 2,3-bis(hydroxymethyl)quinoxaline-1,4-dioxide (dioxidin), which has high bactericidal activity against a wide range of microorganisms. According to the requirements of the State Pharmacopoeia of the Russian Federation XIV edition, related impurities of this API are analysed by thin layer chromatography (TLC). However, this method is semi-quantitative and relative. The purpose of this work is to determination of impurities in the dioxydine substance using high-performance liquid and thin-layer chromatography methods. In accordance with the requirements of the monograph of the State Pharmacopoeia of the Russian Federation, the total content of all impurities in the API is assessed and only the content of quinoxidine is determined separately, which is often absent from the substance. For the first time, three typical dioxidine related impurities were revealed using HPLC. Next, we selected the optimal conditions for their detection by TLC. Methanol/acetonitrile (75/25) and dichloromethane/acetonitrile (50/50) mixtures were the most suitable as a solvent for impurities and the TLC elution system, respectively. Under the selected conditions, satisfactory separation of the chromatographic zones of all four impurities and the dioxidine substance was observed. The corresponding zones were clearly visible at a substance concentration of 1%. However, reducing their concentration to the required 0.1% made it difficult to visually detect impurities. Thus, even when optimizing TLC conditions, it is impossible to obtain reliable data on the content of related impurities, which is due to the limitations of the method. On the contrary, the analysis of dioxidine substances by HPLC allows fairly accurate quantitative determination of individual related impurities.

Comparative Evaluation of the Diuretic Activity of Infusions of Four Species of the Genus Astragalus (<i>Astragalus</i> L.) in the Experiment

Introduction. Astragalus L. is the largest genus of the Fabaceae family and one of the largest genera of vascular plants on Earth. The official type of raw material included in the 7th edition of the European Pharmacopoeia is Radix Astragali – the dried root of Astragalus membranaceus (Fisch.) Bunge and Astragalus mongholicus Bunge. Extracts obtained from astragalus root have anti-inflammatory, immunostimulatory, antioxidant, diuretic effects. In traditional medicine, the herb of Astragalus dasyanthus Pall. is actively used. An infusion of the herb of Astragali dasyanthi has sedative, hypotensive and diuretic effects. However, the raw material is not official and is not included in the list of plant raw materials of the State Pharmacopoeia of the Russian Federation of the XIV edition. Promising species of the genus Astragalus L. can be not Astragalus membranaceus (Fisch.) Bunge and Astragalus dasyanthus Pall., but also other species, for example, Astragalus varius S.G. Gmel., Astragalus testiculatus Pall., Astragalus henningii (Stev.) Klok., which are widespread in the Saratov region. In view of the available data on the presence of the herb of Astragalus dasyanthus Pall. and Astragalus membranaceus (Fisch.) Bunge of the diuretic effect, it is of interest to study the diuresis of extracts from the herb of other species of the genus Astragalus L.Aim. To study the diuretic activity of infusions from the herb of four types of astragalus: Astragalus dasyanthus, Astragalus varius, Astragalus testiculatus, Astragalus henningii in 4- and 24-hour experiments on male rats.Materials and methods. The study of the diuretic activity of aqueous extracts from the herb of four types of Astragalus was performed on 42 white male rats, which were divided into 6 experimental and control groups of 6 animals. Aqueous extracts from Astragalus herb were prepared in the ratio of raw material-extractant 1:10, all obtained infusions met the requirements for these dosage forms according to the General Pharmacopoeia of the State Fund of the Russian Federation of the XIV edition. Aqueous extracts were administered to animals intragastrically through an umbrella at a dose of 100 mg/kg. During the experiment were collected 4-hour and 24-hour portions of urine. The comparison drug in the 4-hour experiment was furosemide at a dose of 1 mg/kg, and in the 24-hour experiment, hydrochlorothiazide at a dose of 20 mg/kg. Results and discussion. As a result of the experiment, the diuretic effect of the infusion of the herb of Astragalus dasyanthus both in the 4-hour experiment and in the 24-hour experiment. Also in both experiments, the most pronounced diuretic effect was found in the infusion of the herb of Astragalus henningii at a dose of 100 mg/kg compared with herb infusions Astragalus dasyanthus, Astragalus varius. Diuretic effect Astragalus henningii exceeded the diuretic activity of furosemide at a dose of 1 mg/kg, while a comparable effect was observed with hydrochlorothiazide at a dose of 20 mg/kg. Diuretic effect of herbal infusion Astragalus varius in a 4-hour experiment exceeded the diuretic effect of herb infusion Astragalus dasyanthus and furosemide at a dose of 1 mg/kg, and in a 24-hour experiment comparable to the diuretic effect of herb infusion Astragalus dasyanthus, but lower than that of hydrochlorothiazide at a dose of 20 mg/kg and herb infusion Astragalus henningii. Diuretic effect of herbal infusion Astragalus testiculatus at a dose of 100 mg/kg was not detected both in the 4-hour and in the 24-hour period. The diuretic activity of the group that received the infusion of the herb of Astragalus testiculatus was at the level of the control group, compared with other experimental groups, a statistically significant decrease in diuresis was observed in animals.Conclusion. The experiment confirmed the diuretic effect of the infusion of the herb of Astragalus dasyanthus and for the first time discovered the diuretic effect of the infusion of the herb of Astragalus varius and the infusion of the herb of Astragalus henningii. Among the studied infusions, the most pronounced diuretic effect was observed in Astragalus henningii herb infusion. The diuretic effect of Astragalus varius infusion is higher than that of Astragalus dasyanthus infusion. in a 4-hour experiment and did not differ in effectiveness from the infusion of the herb of Astragalus dasyanthus in a 24 hour experiment. The diuretic effect of the infusion of the herb astragalus oviparous was not revealed.

Express quantitative spectrophotometric determination of 2-(((3-(2-fluorophenyl)- 5-thio-4H-1,2,4-triazol-4-yl)imino)methyl)phenol as an active substance of a medicinal product for the treatment of mycoses

Foot fungus (Candida, molds, dermatophytes) is a fairly common problem. According to the WHO, one-fifth of the world’s population is affected by fungal skin diseases. The prevalence of mycosis of the feet, which affects every second person, is especially increasing. Such a pathology can appear quite unexpectedly and at an inappropriate time. Today, Ukraine is under aggressive pressure from its northern neighbor, and military personnel are not only constantly at risk of receiving combat injuries and wounds, but also due to active hostilities, they may not have access to the necessary hygiene products, get cold, overheat in the sun, or not have time to change their clothes in time. and shoes. The reality is the absence or delay of treatment; the disease progresses and can spread quickly. If mycosis has appeared in one soldier, it can quickly affect others. Therefore, the problem of providing medical and pharmaceutical care is an urgent issue today. Expanding the assortment of drugs with the above action will ensure the needs of the Armed Forces and civilian patients with this dermatological pathology. It will be able to optimize antifungal therapy regimens. For quite a long time, 1,2,4-triazole derivatives have been a promising class of organic compounds that attract the attention of scientists from various fields. It has been proven that 2-(((3-(2-fluorophenyl)-5-thio-4H-1,2,4-triazole-4-yl)imino)methyl)phenol has high antifungal activity. A potential medicinal product based on this medicinal substance can be used for the treatment of various dermatological diseases. To continue the creation of new effective dosage forms for the effective therapy of mycosis of the feet, the urgent task of today is the development of an express, accurate, reliable, and affordable method of quantitative determination of the substance under study. The aim of the work is to develop an express, sensitive and easy-to-implement UV-method for the quantitative determination of 2-(((3-(2-fluorophenyl)-5-thio-4H-1,2,4-triazole-4-yl)imino)methyl)phenol. Materials and methods. The research object is a working standard sample of 2-(((3-(2-fluorophenyl)-5-thio-4H-1,2,4-triazole-4-yl)imino)methyl)phenol provided by NUNVF, LLC “Brovapharma” according to the “Research and production technological regulations for the production of 2-(((3-(2-fluorophenyl)-5-thio-4H-1,2,4-triazol-4-yl)imino)methyl)phenol” developed based on scientific developed by the Ukrainian Military Medical Academy in conjunction with the National Military Medical Clinical Center “Main Military Clinical Hospital” (term of validity of the regulations until 12/31/2034). Isopropanol of grade “chemically pure” was used as a solvent. Analytical equipment: spectrophotometer Specord 200, electronic scales Radwag XA 210.4Y, class A measuring vessels. Results. An express, eco-friendly spectrophotometric method was developed for the quantification of 2-(((3-(2-fluorophenyl)-5-thio-4H-1,2,4-triazole-4-yl)imino)methyl)phenol in isopropanol area, having analytically maximum at 310 nm. The studied solutions are stable for 30 minutes. The analytical technique is linear in the range of concentrations 0.440–0.754 mg/100 ml (70–130 %). The detection and quantification limits were 5.80 % and 15.70 % respectively. The score was 0.76, according to the “greenness” icon of the analytical technique, using the tools AGREE. The prediction of the total uncertainty of the results of the developed method is 1.70 % (max∆As 2.00 %). The proposed method is relevant according to the requirements of the State Pharmacopoeia of Ukraine. Conclusions. Pharmaceutical development, introduction into industrial production and further prescribing by dermatovenerologists of an effective, safe, and harmless medicinal product will help to expand the range and reduce the level of high import dependence of the domestic market of medicinal products, which will create a more complete and balanced system of medicinal supply in Ukraine and optimize antifungal therapy schemes.

Exploration of surface tension measurement methods for pharmaceutical excipients

Surface tension is a crucial functional indicator for various classes of pharmaceutical excipients, as highlighted in both the Pharmacopoeia of the People’s Republic of China (ChP) < 9601 Guidelines for Functionality-related Characteristics of Pharmaceutical Excipients > and the United States Pharmacopoeia (USP) < 1059 Excipient Performance >. However, there are few systematic studies on surface tension measurement of pharmaceutical excipients, resulting in a lack of stable parameter support in practical applications. In this study, we aim to fill this gap by exploring three different methods for measuring surface tension. These methods were carefully developed taking into account the actual measurement process and statistical theory, thus ensuring their applicability and reliability. Through comparative analyses, we have identified the most suitable measurement methods for different classes of pharmaceutical excipients. In addition, this paper describes the surface adsorption behavior of various excipients. Therefore, this study provides valuable guidance for the determination of surface tension and the study of surface adsorption behavior, which lays the foundation for further comprehensive research in the field of surface tension of pharmaceutical excipients and the improvement of general pharmacopoeia specification.

A Review of Probiotic Ingredient Safety Supporting Monograph Development Conducted by the United States Pharmacopeia (USP)

The United States Pharmacopeia (USP) is an independent, nonprofit science-based organization whose mission is to improve global health through public standards and related products for medicines, food and dietary supplements. Probiotic-based dietary supplements are increasingly popular in the marketplace and USP has developed fourteen monographs specific to probiotic ingredients, including representatives from the Genera Lactobacillus, Bacillus, Streptococcus, and Bifidobacterium. These monographs include the definition of the article, tests for identification, quantification assays (enumeration in the case of probiotics), limits for contaminants, and other quality parameters when appropriate. In addition to quality, the USP also considers the safety of probiotics for monograph development. This report includes an overview of the USP admission evaluation process for probiotics as well as a tabular summary of the probiotic monographs currently available. Pharmacopeia monographs can guide manufacturers and brand owners and protect consumers through establishment of quality standards.

HPLC-UV Analysis of Flavonoid Glycosides in the Herb of &lt;i&gt;Astragalus&lt;/i&gt; (&lt;i&gt;A. dasyanthus&lt;/i&gt;, &lt;i&gt;A. varius&lt;/i&gt;, &lt;i&gt;A. testiculatus&lt;/i&gt;, &lt;i&gt;A. henningii&lt;/i&gt;), Growing in the Volga Region

Introduction. The search for new promising sources of biologically active substances is one of the tasks of modern pharmacognosy. Among the huge variety of species of the genus Astragalus L., only a few of them have been studied. In addition, none of them is included in the current edition of the State Pharmacopoeia of the Russian Federation. In our opinion, the study of four species of this genus of Astragalus L. (A. dasyanthus, A. varius, A. testiculatus, A. henningii) growing in the Volga region is of interest.Aim. Study of the glycoside composition of flavonoids in the astragalus herb of four species (A. dasyanthus, A. varius, A. testiculatus, A. henningii) by HPLC-UV using external standards.Materials and methods. The material was samples of astragalus grass (A. dasyanthus, A. varius, A. testiculatus, A. henningii), collected on the territory of the Saratov region in accordance with the basic rules of collection and preparation and dried to an air-dry state. Aqueous-alcoholic extracts (1 : 50) were analyzed. Qualitative and quantitative analysis of flavonoid glycosides was carried out on an Agilent 1260 chromatograph (Agilent Technologies, USA) equipped with a diode array detector, manual sample injection (Agilent G1328C manual injector, Agilent Technologies USA) and an Agilent Open Lab CDS system for collecting and processing chromatographic data. The elution gradient was selected individually using 0.1 % phosphoric acid/acetonitrile solution as solvents. Compounds were identified by retention times and UV spectra of reference standards (RS). The quantitative content of each detected component was expressed in mg/g of dry raw material. All studies were repeated five times, the results were statistically processed using MS Exсel 2010.Results and discussion. The chromatographic profiles of aqueous-alcoholic extracts from the herbs of four types of astragalus were studied. Differences were found in both the qualitative composition and quantitative content of flavonoid glycosides in the grass of the analyzed species. The largest number of compounds was found in the herb of A. varius (6 compounds), 4 compounds were identified in the herb of A. dasyanthus and A. henningii, and 3 compounds were identified in the herb of A. testiculatus. The dominant components in the herb of A. henningii are orientin and rutin, in the herb of A. testiculatus and A. varius it is cynaroside, and in A. dasyanthus it is rutin.Conclusion. A HPLC-UV method was developed for determining the quantitative content of flavonoid glycosides in extracts from the herbs of four species of astragalus (A. dasyanthus, A. varius, A. testiculatus, A. henningii) and the chromatographic profiles of the analyzed samples were studied.

OBTAINING A WORKING STANDARD SAMPLE OF CHALCONE BUTEINE AND ITS QUANTITATIVE DETERMINATION IN PLANT RAW MATERIALS

Introduction. Butein ‒ 2',4',3,4-tetrahydroxychalcone is found in a number of plants that have a pronounced antimicrobial and antifungal effect: Butea monosperma Lam., hairy string ‒ Bidens pilosa L., tripartite string ‒ Bidens tripartita L., hydrophytum ‒ Hydnophytum formicarum Jack., early thyme ‒ Thymus praecox. Butein is not the dominant representative of the flavonoid class in plants, but it can serve as a marker compound for the analysis of raw materials containing this chalcone. The purpose of the study is to synthesize and analyze a working standard sample of butein, as well as to develop a method for its quantitative determination in Cosmos bipinnate raw materials using solid-phase extraction. Material and methods. Butein synthesis was carried out using the Claisen‒Schmidt reaction, the yield was 44,4%. Identification of the obtained butein was carried out using UV, IR and Mass spectroscopy. Quantitative determination of butein in the raw materials of Cosmos doubly persita was carried out by HPLC in combination with solid-phase extraction. Results. The results obtained confirm that the test sample is butein (2',4',3,4-tetrahydroxychalcone). To determine butein in plant raw materials, a standard sample of this compound should be available, and therefore the substance of this chalcone was synthesized and on its basis, according to the state pharmacopoeia, a working standard sample was obtained (butein content in it is 99,62%). Identification of butein by HPLC was carried out by the retention time, which in the chromatogram of the test solution is 160,70 min, in the chromatogram of the standard solution – 160,07 min  2%. The butein content in the flowers of the cosmos bipinnate we studied in terms of dry raw materials is 0,043%  2%. Conclusions. The synthesis of chalcone ‒ butein was carried out by alkaline condensation of resacetophenone with protocatechuic aldehyde. The yield of butein is 5,95  0,0195 g (44,4%). A working standard sample was obtained based on the butein substance, the authenticity and purity of which was confirmed by melting point, as well as using HPLC, mass spectroscopy, UV and IR spectroscopy. The resulting standard working sample was further used to develop a method for the quantitative determination of butein in Cosmos bipinnate flowers using solid-phase extraction. The butein content in terms of dry raw materials is 0,043%3%.

Control of organic impurities in antibiotics

Introduction: the international, as well as Russian Pharmacopoeia criteria for organic impurity assessment are not applicable to medicinal products obtained by fermentation, therefore the setting of limits for product-related impurities in antibiotics is not regulated by any commonly shared rules. The aim of the study was to analyse and summariese pharmacopoeial requirements for and methodological approaches to organic impurities control in active substances and finished products of semisynthetic antibiotics, peptide antibiotics, and antibiotics obtained by fermentation. Material and methods: the authors analysed the requirements for organic impurities control, as established by the leading world pharmacopoeias and the State Pharmacopoeia of the Russian Federation, using the example of three different groups of antibiotics (semisynthetic antibiotics, antibiotics obtained by fermentation, and peptide antibiotics). The study used comparative analysis and content analysis methods. Results: the study demonstrated that the product-related impurity profiles of the analysed active substances and corresponding finished products often differ significantly across the leading pharmacopoeias, either qualitatively or quantitatively, and do not necessarily comply with the EMA Guideline on setting specifications for related impurities in antibiotics. The methodological approaches to product-related impurity control also differ a lot in terms of pharmacopoeial reference standards used, accelerated stability studies, etc. Conclusion: the complex composition of product-related compounds does not always allow for their unambiguous classification as either impurities or active moieties (e.g. in the case of bleomycin). Most Russian Pharmacopoeia antibiotic monographs are harmonised with the European Pharmacopoeia (Ph. Eur.), which is fraught with problems regarding reference standards (RSs) use, since the composition of Ph. Eur. RSs includes both the active substance and several (sometimes up to 8) impurities. The Russian Pharmacopoeia monographs on antibiotics obtained by a process involving the fermentation stage, include the abnormal toxicity test, which seems redundant in comprehensive impurity control, at least for semisynthetic antibiotics.

Determination of omeprazole via fluorescence-quenching methodology; application to content uniformity testing.

A new, proven, economical spectrofluorimetric approach has been used to determine the proton pump inhibitor omeprazole (OMP). This innovative technique is based on the ability of OMP to quench the native fluorescence of the mercurochrome dye in an acidic (pH3.6) solution. Because it was discovered that quenching is proportional to the drug concentration, this dye was used as a sensor for OMP detection. The fluorescence intensity was measured at 518/540 nm, and its linear response ranged from 0.2-10.0μg/mL with a linear coefficient of 0.9999. The computation yielded a limit of quantification (LOQ) of 0.20 μg/mL and a limit of detection (LOD) of 0.07 μg/mL. Every circumstance and element impacting the reaction product was examined in detail. Pharmacopeial standards carried out the validation. The approved method investigated several commercial preparations and formulations, and the results were favorably compared with those provided by a reference method. According to United States Pharmacopeia (USP) rules, content consistency for two distinct formulations was evaluated.

ПРИМЕНЕНИЕ СОРБЕНТА ДЛЯ ТВЕРДОФАЗНОЙ ЭКСТРАКЦИИ НА ОСНОВЕ ГИПРОМЕЛЛОЗЫ ДЛЯ КОЛИЧЕСТВЕННОГО ОПРЕДЕЛЕНИЯ КИСЛОТЫ САЛИЦИЛОВОЙ В КОМБИНИРОВАННЫХ ЛЕКАРСТВЕННЫХ ПРЕПАРАТАХ

Introduction. In quality control of pharmaceutical formulations, there are methodological approaches based on the preliminary separation of sample components and approaches without preliminary separation. The use of solid-phase extraction (SPE) allows to separate analytes during sample preparation, with the separation selectivity determining the analysis selectivity as a whole. Currently, sorbents that provide selectivity in interaction with the analyte are of interest. Various derivatives of cellulose are used to obtain selective sorbents: ethylcellulose, hydroxypropylmethylcellulose (hypromellose), carboxymethylcellulose, etc. On the pharmaceutical market of the Russian Federation, there are pharmaceutical formulations with the active pharmaceutical substance salicylic acid. The use of selective SPE for analysis is a promising direction of research in drug quality control. Aim. Evaluation of the metrological characteristics of benzocaine quantitation by UV-spectrophotometry in certain pharmaceutical formulations using hypromellose sorbent for solid-phase extraction. Material and methods. The sorbent using a method developed by us was obtained. Salicylic acid (FS.2.1.0634, Russia State Pharmacopoeia XV ed.) to form active binding sites in sorbent structure was applied. The sorbent structure is polycyanoacrylate matrix with hypromellose fragments, the surface area is 255.50 m2/g, pore volume is 0.1433 cm3, pore diameter is 5.32 nm. The sorption capacity of hypromellose sorbent for benzocaine was 12.2 ± 0.8 μg/g. The SPE technique is proposed. SPE includes the stages: conditioning, sample addition and step-by-step elution with purified water and sodium hydroxide solution 0.1 mol/L. The spectrophotometer SF-56 was used to measure the absorbance at an analytical wavelength of 286 nm. To evaluate the selectivity (specificity) of sample preparation during chromatographic (SPE) separation of pharmaceutical formulations, absorption spectra of eluates in the wavelength range of 200-400 nm were scanned. The salicylic acid identification in the eluate was carried out based on absorption peaks at 228 and 298 nm.The salicylic acid calibration curve in the range of 1-20 μg/ml. To determination the metrological characteristics of salicylic acid quantitation method, 16 parallel determinations of samples of each pharmaceutical formulation were made. Statistical processing of the experimental results and determination of the metrological characteristics evaluation of analytical technique were carried out in accordance with general pharmacopoeial monograph (OFS.1.1.0013 "Statistical processing of results of physical, physicochemical, and chemical tests"). Results. When realizing the salicylic acid quantitation technique for measuring absorbance, it is advisable to use only the first portions of eluate (a solution of sodium hydroxide) in a volume of 5-10 ml were obtained. The relative error in determining the average content of salicylic acid in the analyzed preparations ranged from 1.18 to 1.44% for methods with solid-phase extraction (SPE) and 1.86-2.62% in its absence. It has been shown that there is a statistically significant difference in reproducibility for all preparations, as indicated by F-criteria values ranging from 7.81 to 13.5, exceeding their tabulated values. For the method without solid-phase extraction (SPE), there was an overestimation of the salicylic acid content in all cases, increasing the systematic error on average by 87.5%. Conclusion. The possibility of using a selective hypromellose sorbent for sample preparation for salicylic acid quantitation determination in pharmaceutical formulations has been showed. The use of sorbent for analysis provides to systematic error decrement of salicylic acid quantitation by an average of 87.5%.

Composition and technology development of ear drops with cerumenolytic action (based on thick Viscum album L. leaves extract)

Earwaxes lead to a decrease in the quality of life. A rational and effective way to eliminate earwaxes without a participation of medical personnel is to use ear drops and solutions to dissolve sulfur masses. In this regard, therapeutic and hygienic drops of а cerumenolytic action are of a particular relevance in the pharmaceutical market.The aim of the work was to develop a composition and technology of ear drops of a cerumenolytic action based on thick Viscum Album L. leaves extract.Materials and methods. The objects of the study were thick Viscum Album L. leaf extract, chitosan, sodium alginate, polyethylene oxide of various degrees of polymerization, propylene glycol, sodium hyaluronate, preservatives (benzalkonium chloride, nipagine and nipazole). At the screening stage, 9 experimental formulations were proposed. The cerumenolytic activity of the developed formulations was evaluated in a dissolution test of reproduced artificial earwax in comparison with a 3% hydrogen peroxide solution, TEA-cocoyl hydrolyzed collagen (A-Cerumen Plus, Gilbert Laboratories, France) and a 0.9% sodium chloride solution. Physical and chemical parameters (a degree of liquid coloring, turbidity and transparency, pH, density and viscosity) were determined according to of the State Pharmacopoeia of the Russian Federation (XV ed.). The microbiological study was performed using the agar diffusion method.Results. In the course of the study, it was shown that the composition of ear drops of a cerumenolytic action based on thick Viscum Album L. leaves extract, exceeds the level of a lipolytic, proteolytic and general cerumenolytic kinds of activity of the compositions containing surfactants; in its effectiveness, it was comparable with the comparison drug – A-Cerumen drops. To obtain the optimal rate of the onset effect and its duration, it is advisable to use sodium hyaluronate in the amount of 0.2 g per 25 ml of drops as an adjuvant. The most preferred preservative was benzalkonium chloride. The developed ear drops met the requirements of the State Pharmacopoeia of the Russian Federation (XV ed.) for this dosage form, while the pH was 5.86± 0.1, and the viscosity was 4.2676±0.2 MPa×s.Conclusion. The conducted research has shown the prospects for further work on the development and implementation of ear drops of a cerumenolytic action in practice. The recommended composition is the following: thick Viscum Album L. leaf extract, sodium hyaluronate, benzalkonium chloride, purified water.

APPLICATION OF SELECTIVE HYPROMELLOSE SORBENT FOR THE BENZOCAINE QUANTITATION IN PHARMACEUTICAL FORMULATIONS

Introduction. In quality control of pharmaceutical formulations, there are methodological approaches based on the preliminary separation of sample components and approaches without preliminary separation. The use of solid-phase extraction (SPE) allows to separate analytes during sample prepa-ration, with the separation selectivity determining the analysis selectivity as a whole. Currently, sorbents that provide selectivity in interaction with the analyte are of interest. Aim. Evaluation of the metrological characteristics of benzocaine quantitation by UV-spectrophotometry in certain pharmaceutical formulations using hypromellose sorbent for solid-phase extraction. Material and methods. The sorbent using a method developed by us was obtained. Benzocaine (FS.2.1.0634, Russia State Pharmacopoeia XV ed.) to form active binding sites in sorbent structure was applied. The sorbent structure is polycyanoacrylate matrix with hypromellose fragments, the sur-face area is 255.50 m2/g, pore volume is 0.1433 cm3, pore diameter is 5.32 nm. The sorption capacity of hypromellose sorbent for benzocaine was 12.2±0.8 μg/g. The SPE technique is proposed. SPE includes the stages: conditioning, sample addition and step-by-step elution with purified water and hydrochloric acid solution 0.1 mol/L. The spectrophotometer SF-56 was used to measure the absorbance at an analytical wavelength of 286 nm. To evaluate the selectivity (specificity) of sample preparation during chromatographic (SPE) separation of pharmaceutical formulations, absorption spectra of eluates in the wavelength range of 200400 nm were scanned. The benzocaine identification in the eluate was carried out based on absorption peaks at 220 and 286 nm. The benzocaine calibration curve in the range of 120 μg/ml. To determination the metrological characteristics of benzocaine quan-titation method, 11 parallel determinations of samples of each pharmaceutical formulation were made. Results. When realizing the benzocaine quantitation technique for measuring absorbance, it is advisable to use only the first portions of eluate (a solu-tion of hydrochloric acid) in a volume of 510 ml were obtained. The relative error of benzocaine average concentration in the pharmaceutical formula-tions ranged from 1.28 to 1.34% for technique that included the SPE stage, and from 1.86 to 2.02% for technique that did not include it. A comparison of the modifications of the spectrophotometric quantitation of benzocaine in pharmaceutical formulation using hypromellose sorbent and technique without the SPE did not reveal a statistically significant difference in reproducibility. For the modification technique without SPE, an increase of benzo-caine concentration and in determination relative error by an average of 48.0% were observed. Conclusion. The possibility of using a selective hypromellose sorbent for sample preparation for benzocaine quantitation determination in pharmaceu-tical formulations has been showed. The use of a sorbent for SPE at the sample preparation stage reduces the systematic error of benzocaine spectro-photometry quantitation for the analyzed pharmaceutical formulations by an average of 79.2%.

Disintegration testing for orally disintegrating tablets (ODTs): An overview

Prescribing orally disintegrating tablet (ODT) dosage form is a common practice. For several populations, including geriatrics and pediatrics, ODTs are one of the most desired dosage forms. They are solid dosage forms designed to disintegrate or dissolve very quickly in contact with saliva as soon as they are placed in oral cavity of a patient. According to United States Pharmacopeia (USP), every dosage form has to pass some quality control tests to ensure uniformity and performance of this dosage form. For ODTs, disintegration test is fundamental to ascertain time required for tablets to break down and release their contents for dissolution and absorption. Additionally, it is a crucial predictive test for determining in vitro/in vivo association. There are no mandatory universal disintegration testing standards for ODTs, however, as stated in the USP. Instead, recent USP refers to individual manufacturer monograph specifications which may vary from one monograph to another. This article discusses several developed disintegration tests and methods for ODTs such as OD-mate testing, basket rack assembly, CCD camera, texture analyzer (TA), special disintegration equipment, prototype disintegration tester (PDT), simple approach, and modified wetting test and elaborates on their advantages and disadvantages.

Острые респираторные вирусные инфекции: фокус на тяжесть течения. Блиц-обзор

This concise examination of international and domestic research on the impact of respiratory viruses and age-specific characteristics of the immune response in young children on the severity of influenza and other acute respiratory viral infections (ARVIs) is designed to highlight the importance of timely and effective therapy for these diseases. When selecting therapy for ARVIs in children, particularly those of a younger age, it is advisable to adopt a balanced approach that ensures adequate antiviral protection and rapid elimination of the virus, while also considering the necessity for effective and safe therapy to prevent an inadequate immune response. Furthermore, the review addresses the findings on a representative of the group of "Biological medicines produced using gradual processing technology" (State Pharmacopoeia of the Russian Federation, XV ed.), namely the domestic antiviral drug Anaferon® for children in drops for oral administration. The drug has been demonstrated to have antiviral activity, to provide a significant effect on reducing the duration of fever, and the severity of the disease course, and to have the favorable safety profile. Additionally, it is administered in a convenient, physiological manner, allowing for its wide application in the treatment of acute respiratory infections of the upper respiratory tract in children from 1 month to nearly 4 years (3 years, 11 months, and 29 days inclusive). KEYWORDS: ARVI, acute respiratory viral infection, acute respiratory infection of the upper respiratory tract, common cold, severity of course, treatment. FOR CITATION: Zaplatnikov A.L., Geppe N.A., Blokhin B.M., Kondyurina E.G. Acute respiratory viral infections: focus on severity. A brief overview. Russian Journal of Woman and Child Health. 2024;7(3):270–275 (in Russ.). DOI: 10.32364/2618-8430-2024-7-3-11.

A new ultrasound-assisted microextraction for elemental impurities determination in tablets by ICP-MS according to USP chapters 232 and 233

Along with the United States Pharmacopeia (USP) chapters 232 and 233 regarding elemental impurities in pharmaceutical products, new challenges have been imposed in terms of sample preparation procedures prior to inductively coupled plasma mass spectrometry analysis, considering the matrix complexities. As so, a new microextraction procedure assisted by ultrasound using a cup-horn sonoreactor, minimal reactants, and sample was proposed and validated according to USP. The procedure was optimized with samples of milled tablets and 3 different acid mixtures (HNO3, 3HNO3:1HCl, and 9HNO3:1HF) and it was compared with microwave-assisted acid digestion. In the validation step, recoveries ranging from 85 to 120 % and RSD below 10 % were obtained for 22 analytes (except Ag and Pt) with satisfactory linearity and good sensitivity. The method was then applied for 37 samples of antidepressants, which presented trace levels of As, Ba, Cd, Co, Cr, Cu, Ni, Pb, Pd, Sn, and V.