Inter-Laboratory Comparison of Photometric and LC Assay Methods for Hydroxyanthracene Glycosides Determination in Senna Herbal Drugs and Dry Extracts.

Aim. In vitro evaluation of the inhibitory activity of aqueous extracts and dry ethanol extracts of St John's wort (Hypericum perforatum L.) on the replication of SARS-CoV-2 according to three experimental schemes – direct inactivation (neutralisation) of the virus as well as "prevention" and "treatment" of cells.Materials and Methods. The laboratory strain SARS-CoV-2/human/RUS/Nsk-FRCFTM-1/2020 was passed on Vero cell culture. Water extracts and dry ethanol extracts of parts of H. perforatum L. collected during the flowering period in the Novosibirsk region were prepared. Dry extracts were dissolved in DMSO. Comparison samples are dry ethanol extracts of chaga, cloves and black tea.Results. It is shown that the aqueous extract of grass (a mixture of flowers with leaves) of H. perforatum L. with direct inactivation of the virus it is active in dilution of 1/4096. For the dry ethanol extract of the herb H. perforatum L., 50 % effective concentrations (EC50) were found equal to 2.44±0.87; 8.79±1.91 and 14.65±1.91 μg/ml respectively with direct inactivation as well as according to the "preventive" scheme and with the "treatment" of cells. Taking into account cytotoxicity, as well as in comparison with control samples, the values of selective indices (SI50) of the studied herbal preparations during direct inactivation were higher than with other experimental schemes and were distributed as follows (in descending order): 204.92; 153.68; 115.27; 32.01 and 21.33 for dry ethanol extracts of black tea from India, cloves, herbs, a mixture of flowers with leaves, of H. perforatum, chaga and the stems of H. perforatum respectively. The HPLC method has shown that the ethanol extract of the herb H. perforatum L. contains a greater amount of flavonoids than the extract of stems. Nevertheless, antiviral activity was also detected for the extract of stems of this plant with EC50 equal to 14.65±1.91; 78.13±20.05 and 117.19±15.31 μg/ml (according to three experimental schemes), respectively.Conclusion. For the preparation of antiviral drugs the whole plant of H. perforatum L., including stems, can be used as raw materials.

Clinical performance characteristics of a new photometric lithium assay: a multicenter study

A colorimetric and ratiometric photometric sequential assay for ascorbic acid and alkaline phosphatase in serum based on valence states modulation

Using standard laboratory equipment a statistical comparison was carried out of several photometric methods (normal and various Higuchi plots) and potentiometric methods (point of inflection and Gran) for equivalence-point location in an acetic acid-sodium hydroxide solution titration. All methods exhibited similar precision but were divided into two groups with respect to accuracy. With increasing carbonate concentration in the titrant there was a decrease in the linear range for Higuchi and Gran plots and a decrease in accuracy for all methods.

Assay of total factor XII and of activated factor XII in plasma with a chromogenic substrate

A new photometric method is presented for the assay of serum glutamate dehydrogenase activity on the basis of the determination of 2-oxoglutaric acid produced in the enzyme reaction by means of the previously established method for selective determination of the acid with diazotized sulfanilic acid. The method gives reliable results and is suitable to assay a large number of samples at the same time with 0.1 ml of sample.

Evaluation of a Sensitive Colorimetric FXIII Incorporation Assay. Effects of FXIII Val34Leu, Plasma Fibrinogen Concentration and Congenital FXIII Deficiency

The EU Response to COVID-19: From Reactive Policies to Strategic Decision-Making.

In this study, fibrinogen measurements according to the Clauss method, photometric method and Jacobsson method have been investigated to find out how they are influenced by adding in vitro the purified canine fibrinogen degradation products (FDP) X, Y and D. Test results according to the Clauss method were found to be underestimated if the fragments X, Y and D were added while measurements according to the Jacobsson method turned out to underestimate the real fibrinogen concentration if the FDP Y and D were added. The Clauss method was particularly sensitive towards FDP. Results were considerably underestimated even with a quantity as little as 0.05 g FDP Y or FDP D/g fibrinogen (p < 0.05). The photometric method was only affected by FDP X leading to false high results. If FDP X was added, fibrinogen values were also overestimated with the Jacobsson method. Our results demonstrate that the photometric method is the most accurate.

BackgroundClinical study reports (CSRs) have been increasingly utilised within academic research in recent years. European Medicines Agency (EMA) Policy 0070 ‘Phase 1,’ which came into effect in January 2015, requires the publication of regulatory documents such as CSRs from central applications in an anonymised format. EMA Policy 0070 requires sponsors to demonstrate careful consideration of data utility within anonymised CSRs published within the scope of the policy, yet the concept of data utility is not clearly defined in the associated anonymisation guidance.ObjectiveTo review the use of data from CSRs in published academic research and to hypothesise the potential data utility of CSRs anonymised under the objectives of EMA Policy 0070 for future academic research.MethodsReview of the objectives, research methodologies and findings of academic research reports using unpublished data from CSRs (prior to EMA Policy 0070). Semi-structured interviews with authors of academic research reports, including questions related to data utility of anonymised CSRs published under EMA Policy 0070.ResultsThirteen academic research reports were identified and reviewed. The research purposes ranged from assessment of reporting bias, comparison of methods and results with published data sources, detailed evaluation of harms and adverse events, re-analysis and novel analyses including systematic reviews and meta-analysis. All of the examples identified required access to the methods and results sections of CSRs (including aggregated summary tables) and research purposes relating to evaluation of adverse events also required access to participant narratives. Retaining anonymised participant narratives relating to interventions, findings and events, while maintaining an acceptably low risk of participant re-identification, may provide an important gain in data utility and further understanding of drug safety profiles.ConclusionsThis work provides an initial insight into the previous use of CSR data and current practices for including regulatory data in academic research. This work also provides early guidance to qualitatively assess and document data utility within anonymised CSRs published under EMA Policy 0070.

Plasma fibrinogen concentration was measured in 67 healthy, adult dogs using five different methods (gravimetry, methods described by JACOBSSON (1955) RATNOFF and MENZIE (1951), and CLAUSS (1957), and functional photometric assay). Apart from using linear regression and the Pearson correlation coefficient (r) in order to characterize the relation between different methods, reference ranges (2.5-97.5%-fractile) were calculated for all methods. For calibration of the CLAUSS method (1957) and the photometric assay, dog plasma with a defined fibrinogen concentration was used. Measurements of commercial human fibrinogen standards yielded a good conformity with the concentrations specified by the manufacturer (values approximately 3% too low). These standards appear, therefore, to be also suited to the calibration of measurements of dog fibrinogen. The reference range for the gravimetry was 1.08-2.88 g fibrinogen per litre of plasma. A considerable conformity and close correlation was seen between the fibrinogen concentration measured by gravimetry and by using methods described by JACOBSSON (1955; y = 1.088 x -0.142, r = 0.967) or CLAUSS (1957; y = 0.999 x -0.004, r = 0.973), respectively. Between the reference-method gravimetry and the photometric method and RATNOFF-MENZIE (1951) method, respectively, a less close correlation, as well as a minor conformity, was found.

Specific effects of ATP on the kinetics of reconstituted bovine heart cytochrome- c oxidase

Small-molecule inhibitors have transformed oncology in recent years. This study compared regulatory approvals of these agents across the United States Food and Drug Administration (FDA), the European Medicines Agency (EMA) and the Japanese Pharmaceuticals and Medical Devices Agency (PMDA), focusing on timelines, dosing recommendations, and pediatric labelling. A review of regulatory databases was conducted to identify small-molecule inhibitors approved for adult malignancies. Drug labels were compared to determine dosing concordance (Fully, Partially, or Non-Concordant), approval dates, and pediatric indications. Data extraction involved two independent reviewers. Fifty-five inhibitors were approved by all three agencies. Adult dosing was Fully Concordant in 49 (89%), partially in 4 (7%), and Non-Concordant in 2 (4%). The median approval gap was 25months (range: 1-88). FDA granted first approval for 85.5% of agents, followed by PMDA (12.7%) and EMA (1.8%). Among these 55 drugs, only 15 had pediatric indications (27%), 7 of them (46.7%) approved across all three regions. No complete divergence in pediatric dosing was observed, although minimum age thresholds varied. Despite strong alignment in adult dosing, regulatory disparities in approval timelines and pediatric labelling persist, risking delays in therapy availability. More harmonized multinational trials and regulatory alignment could facilitate timely approvals while allowing for population-specific considerations in dosing and safety.

An HPLC method and an acid hydrolysis HPLC method for the analysis of anthocyanins and anthocyanidins in bilberry extracts have been developed. The HPLC method coupled with a mass detector has identified 11 anthocyanins in bilberry extracts. The method provides anthocyanin profiles that are very useful in verifying the identity of botanical raw materials, monitoring the consistency of the raw material source, and quantitating the total anthocyanins. The acid hydrolysis HPLC method greatly simplifies the anthocyanin profile in bilberry samples and converts anthocyanins to five major anthocyanidin aglycones: delphinidin, cyanidin, petunidin, peonidin, and malvidin. Each of these aglycones can be separated completely and quantitated accurately with external standards. Various extraction and hydrolysis conditions were investigated, and the advantages and disadvantages of the HPLC and acid hydrolysis methods are discussed.

In the world pharmaceutical practice, there are significant differences regarding the requirements for the technological parameters of aqueous extracts from medicinal plant raw materials. Aim. To conduct a comparative analysis of the requirements of regulatory documents of Ukraine, the European Union and Poland regarding the compounding of water extracts from medicinal plant raw materials. Materials and methods. Comparative analysis was carried out using systemic and structural-logical analysis. The object of research was normative legal acts regulating medicines compounding in Ukraine, European Pharmacopoeia (EP) XI ed., European Medicines Agency (EMA), Pharmacopoeia of Poland (PP) VI and IX ed., textbook "Farmacja stosowana, Technologia formaci leku; podręcznik dla studentów farmacji" and others. Results and discussion. As a result of the analysis, a few distinctive features in the technology of water extracts from medicinal plant raw materials were revealed compared to the requirements of the EU and Poland. According to the requirements of European Pharmacopoeia and the European Medicines Agency, aqueous extracts from medicinal plant raw materials, so-called herbal teas, are prepared with the methods of maceration, infusion and decoction. Macerates are prepared from mucilaginous medicinal plant raw materials with the method of cold infusion at room temperature for 30 minutes. Infusions are usually prepared from delicate parts of the plant, while medicinal plant raw materials are poured with hot water with an initial temperature of 90-100 ºС and infused by gradually cooling the extract at room temperature for 5-15 minutes. Decoctions are usually prepared from roots, rhizomes and bark by boiling medicinal plant raw materials in a boiling water bath for 15-30 minutes, while the raw materials are initially poured with an extractant at room temperature, and after boiling, the extract is immediately filtered. The compounding peculiarities of water extracts from medicinal plant raw materials in Poland are as follows: the ratio of medicinal plant raw materials to the extractant with weak biologically active substances is 1:10, with mucus-containing medicinal plant raw materials – 1:20; for medicinal plant raw materials with cardiac glycosides and alkaloids – 1:100; water extracts are prepared by mass, while water absorption coefficients or consumption coefficients are not taken into account; for acidification, 0.5 g of citric acid per 100 g of extractant is added to extracts from alkaloid-containing raw materials; macerates are prepared from mucilage-containing raw materials only by the cold infusion method (for example, flax seed macerate); infusions are prepared only from medicinal plant raw materials containing cardiac glycosides (Lily of the Valley herb, Adonis vernalis herb), where raw materials are poured with boiling water, infused in a water bath for 15 minutes and cooled at room temperature for 15 minutes; decoctions are always prepared from other types of medicinal plant raw materials, where the raw materials are poured with an extractant at room temperature, heated and boiled in a boiling water bath for 30 minutes, filtered without cooling. Conclusions. A comparative analysis of the technological parameters of water extracts compounding from medicinal plant raw materials in accordance with the requirements of regulatory documents of Ukraine, the European Union and the Republic of Poland was carried out. A number of differences have been established, particularly in the methods of recipes prescribing, features of calculations, in the ratio of medicinal plant raw materials to the extractant, temperature regimes, extraction time, medium pH, the concepts of the processes of maceration, infusion and decoction. It was found that compared to domestic regulations, the technological process in the EU and Poland is optimized in terms of time, energy and in some cases, raw material expenditure. The obtained results have a practical importance, as they are the basis for further scientific research on the substantiation of the influence of technological factors on the quantitative content of the main groups of biological active substances in the compounded water extracts.