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        <title>Frontiers in Analytical Science | Pharmaceutical analysis section | New and Recent Articles</title>
        <link>https://www.frontiersin.org/journals/analytical-science/sections/pharmaceutical-analysis</link>
        <description>RSS Feed for Pharmaceutical analysis section in the Frontiers in Analytical Science journal | New and Recent Articles</description>
        <language>en-us</language>
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        <pubDate>2026-05-13T15:07:06.382+00:00</pubDate>
        <ttl>60</ttl>
        <item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2026.1739382</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2026.1739382</link>
        <title><![CDATA[Capillary gel electrophoresis for the analysis of heterodimer purity in a highly glycosylated, bispecific Fc fusion protein]]></title>
        <pubdate>2026-03-12T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Caroline Daniels</author><author>Kyle Jones</author><author>Vishruti Makani</author><author>Cole Schmidt</author><author>Joseph Pate</author><author>Dan Wu</author><author>Taylor H. Schreiber</author><author>Suresh De Silva</author><author>Abhinav Shukla</author><author>Nathan Oien</author>
        <description><![CDATA[IntroductionBispecific therapeutic fusion proteins require analytical methods capable of detecting and resolving process- and product-related impurities while remaining practical to implement and validate. Such methods are essential for supporting both process development and good manufacturing practice (GMP) -compliant testing.MethodsA sodium dodecyl sulfate capillary gel electrophoresis (CE-SDS) method was developed to separate structurally similar protein species based on differences in the number of N-linked glycosylation sites. The approach leverages glycosylation-dependent electrophoretic mobility to resolve heterodimeric and related molecular species.ResultsThe CE-SDS method effectively separated bispecific protein variants that differed in glycosylation, enabling discrimination between the desired bispecific product and associated impurities. This was confirmed by comparing native and deglycosylated Fc-fusion protein. The method was successfully applied to inform process development decisions aimed at enriching the target bispecific protein and was demonstrated to be suitable for GMP release and stability testing by verifying ICH criteria for method validation.DiscussionThis analytical strategy provides a robust and broadly applicable means of separating heterodimeric and bispecific glycoproteins based on glycosylation differences. The method can be readily extended to other fusion proteins, provided sufficient glycosylation asymmetry exists between constituent polypeptide chains, making it a valuable tool for both development and manufacturing control.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2026.1772719</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2026.1772719</link>
        <title><![CDATA[A case study on N-nitrosamine investigation in a marketed drug: from analytical procedure validation to risk mitigation decision]]></title>
        <pubdate>2026-03-12T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Yue Zhang</author><author>Sabah Houari</author><author>Thomas Van Laethem</author><author>Priyanka Kumari</author><author>Eric Ziemons</author><author>Philippe Hubert</author><author>Cédric Hubert</author>
        <description><![CDATA[IntroductionN-nitrosamines, a class of organic compounds recognized for their mutagenic and carcinogenic properties, have been raising major public health concerns following their detection in pharmaceuticals since 2018. These impurities can form during manufacturing processes, particularly through the interaction of amines with nitrosating agents. Regulatory bodies have launched procedures mandating a systematic investigation of medicinal products for N-nitrosamine contamination. This process involves risk evaluation, confirmatory testing, and the implementation of risk mitigation measures. Hyphenated analytical techniques, such as liquid chromatography-tandem mass spectrometry (LC-MS/MS), have become the standard for N-nitrosamine detection thanks to their high sensitivity and specificity.MethodsIn this work, based on a previously developed Quantitative Structure Retention Relationship approach for the determination of N-nitrosamines, sensitive LC-MS/MS methods were proposed and validated. These methods were subsequently applied to confirmatory testing of a marketed drug product.ResultsThe methods were successfully validated in accordance with the ICH Q2 (R2) guidelines as limit tests for trace-level detection of N-nitrosamines. The confirmatory testing demonstrated the absence of most N-nitrosamines throughout the product’s shelf life, except for one potentially problematic impurity that nevertheless remained below the established acceptable limit, thus requiring routine quantitative analysis.DiscussionThese findings highlight the need for continued monitoring through routine analysis to ensure the safety and quality of pharmaceutical products.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2026.1735125</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2026.1735125</link>
        <title><![CDATA[The use of QbD for the development and validation of a stability-indicating RP-HPLC method for the quantitation of nevirapine in bulk, tablet and niosome formulations]]></title>
        <pubdate>2026-02-03T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Bwalya A. Witika</author><author>Roderick B. Walker</author>
        <description><![CDATA[IntroductionA reversed-phase high-performance liquid chromatographic (RP-HPLC) method was developed and validated for the quantitation of nevirapine (NVP) in bulk drug, commercial tablets, and niosome formulations using an Analytical Quality by Design (AQbD) approach.MethodsCritical analytical attributes and method parameters were identified and optimized using a Central Composite Design (CCD), with the retention time and resolution between NVP and internal standard carbamazepine as key responses. Chromatographic separation was achieved on a C18 column using an isocratic mobile phase of water and acetonitrile (57.5:42.5 v/v), a 1.0 mL/min flow rate, and detection at 280 nm. The method was validated per ICH Q2 (R1) guidelines for all parameters including repeatability, intermediate precision, accuracy (recovery/bias), and robustness, in addition to specificity, linearity, LOD and LOQ.Results and DiscussionThe method demonstrated specificity, linearity (0.5–200 μg/mL), a detection limit of 0.033 μg/mL, and quantification limit of 0.5 μg/mL. The method was precise, accurate, and robust. Stress studies suggested stability-indicating performance under the tested conditions with no observed interference at the analyte retention time at 280 nm. Application to commercial and in-house formulations confirmed its suitability for routine analysis. This work highlights the value of AQbD in developing cost-effective, high-performing analytical methods for pharmaceutical analysis.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2025.1533486</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2025.1533486</link>
        <title><![CDATA[The chemicalome profiling of Zishen Yuzhen Pill in vivo and its promoting effect on osteogenic differentiation of MC3T3-E1 cells]]></title>
        <pubdate>2025-02-19T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Juanjuan Cheng</author><author>Xinyue Meng</author><author>Daozheng Fang</author><author>Yong Zhu</author><author>Zhihao Liu</author><author>Xinyue Li</author><author>Ke Jie</author><author>Shiying Huang</author><author>Huilin Li</author><author>Shangbin Zhang</author><author>Jihang Chen</author><author>Jianping Chen</author>
        <description><![CDATA[Zishen Yuzhen Pill (ZYP) is a Chinese herbal product developed by Shenzhen TCM Hospital, which have been frequently used to treat osteoporosis (OP). This study aimed to determine the major chemical components of ZYP and its prototype compounds and metabolites in rat biological samples, as well as explore the potential effect of ZYP-containing serum in MC3T3-E1 cells. UPLC-Q/TOF-MS was used to identify the chemical components. Then, ZYP was orally administered to rat, and samples of plasma, urine, feces, bile, and tissue were collected to identify prototype compounds and metabolites. The viability of MC3T3-E1 cells was evaluated using the CCK-8 method after treatment with various concentrations (2%, 4%, and 8%) of ZYP-containing serum. Following treatment of MC3T3-E1 cells with ZYP-containing serum, the activity of alkaline phosphatase (ALP) and Alizarin red S (ARS) were measured, and the levels of Runx2, Opn, Opg and Osterix were quantified using the qPCR and Western blot analysis. And cells were collected for RNA-seq analysis. Results indicated that a total 152 compounds were identified in ZYP, including flavonoids, iridoid, lignans, triterpene saponins, etc. Furthermore, we detected a total of 70 prototype components and 99 metabolites distributed in different tissues. In addition, ZYP-containing serum observably promoted osteogenesis by increasing ALP and ARS activities, as well as up-regulating the expression of Runx2, Opn, Opg and Osterix in MC3T3-E1 cells. RNA-seq results indicated that the beneficial effects may be related to the upregulation of mitochondrial oxidative phosphorylation. This work provided further support for the traditional application of ZYP in the treatment of OP. And this study can promote the further pharmacokinetic and pharmacological evaluation of ZYP.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2024.1490093</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2024.1490093</link>
        <title><![CDATA[The clinical applications of drugs and their metabolites analysis in biological fluids and commonly used analytical techniques for bioanalysis: review]]></title>
        <pubdate>2024-11-26T00:00:00Z</pubdate>
        <category>Review</category>
        <author>Biset Asrade Mekonnen</author><author>Muluabay Getie Yizengaw</author><author>Kebede Feyisa Adugna</author>
        <description><![CDATA[BackgroundThis study provides an overview of the clinical applications of drugs and their metabolites analysis in biological fluids and identifies commonly used analytical techniques for bioanalysis.MethodsOriginal open-access articles published between 31 October 2005 and 31 October 2020 in Google Scholar, MEDLINE, PubMed, and Embase were reviewed, and pertinent findings of the individual studies were pooled and presented using tables. This review was reported according to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA).ResultsFifteen studies met the eligibility criteria and were included in the review. These studies show that qualitative identification and quantitative determination of drugs and their metabolites in biological fluids are important for therapeutic drug monitoring (TDM), pharmacokinetic and pharmacodynamic studies, drug abuse control, and forensic and toxicological analyses. Spectroscopic, electrochemical, and hyphenated and nonhyphenated chromatographic techniques are used to analyse drugs and their metabolites in biological fluids. However, hyphenated techniques are the preferred analytical methods because of their sensitivity, selectivity, accuracy, reproducibility, efficiency, and rapid analysis.Conclusion/RecommendationsBioanalysis is important for pharmacokinetic and pharmacodynamic studies, therapeutic drug monitoring, forensic and toxicological analyses, and drug abuse control using different bioanalytical techniques. However, hyphenated techniques are the most commonly used bioanalytical techniques. Bioanalysis of drugs and their metabolites needs to be improved to provide good medical and pharmaceutical care to patients, to confirm forensic and toxicological cases, and to control drug abuse.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2024.1498404</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2024.1498404</link>
        <title><![CDATA[Ultrasound-assisted extraction of itraconazole from pellets: an efficient approach for the recovery of active pharmaceutical compounds from solid dosage forms]]></title>
        <pubdate>2024-10-30T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Patricia G. Ferreira</author><author>Carolina G. S. Lima</author><author>Marcela C. de Moraes</author><author>Fernando de C. da Silva</author><author>Débora O. Futuro</author><author>Vitor F. Ferreira</author>
        <description><![CDATA[IntroductionItraconazole is a widely used broad-spectrum antifungal drug on a global scale. However, its poor water solubility necessitates the development of advanced formulations for its effective use. Currently, itraconazole is available commercially in capsule form, with pellets containing the active drug. The extraction of itraconazole from these granules is often required when developing new formulations. Extracting active pharmaceutical ingredients (APIs) from pellets is crucial in ensuring precise and consistent dosing when producing pharmaceutical dosage forms. Surprisingly, no existing methods for extracting itraconazole from pellets have been reported thus far.ObjectiveTherefore, this study aimed to develop and evaluate an efficient method for extracting this API from pellets.MethodsTwo extraction methods were assessed: conventional Soxhlet extraction and ultrasound-assisted extraction.Results and DiscussionThe results demonstrated that ultrasound-assisted extraction significantly outperformed the conventional method, yielding higher amounts of itraconazole with a high purity level of 96.8%.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2024.1399843</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2024.1399843</link>
        <title><![CDATA[In vitro comparative quality assessment of different brands of hydrochlorothiazide tablets marketed in Northeast Ethiopia]]></title>
        <pubdate>2024-07-30T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Yehualashet Teshome Wondmkun</author><author>Haile Kassahun Desta</author><author>Yimer Seid Ali</author><author>Abate Wondesen Tsige</author><author>Kassahun Dires Ayenew</author><author>Bedilu Linger Endalifer</author><author>Samuel Berihun Dagnew</author>
        <description><![CDATA[BackgroundThis study aimed to comparatively evaluate the quality and physicochemical bioequivalence of commercially available brands of hydrochlorothiazide (HCT) tablets in community pharmacies in Dessie town, Northeast Ethiopia.MethodsExperimental cross-sectional study design was applied using pharmacopeia and non-pharmacopeia methods. The difference (f1) and similarity (f2) factors were calculated to assess the in vitro bioequivalence of generic products with the comparator.ResultThe study results revealed that all investigated brands contained the required active pharmaceutical ingredients (APIs). The friability test results were concordant with the United States Pharmacopeia (USP) (<1%) for eight brands; however, brand NF3 (1.36%) failed to pass the specification limit. The hardness levels of the brands NF3 (24.20 ± 7.32 N), NF5 (32.19 ± 4.78 N), and NF9 (35.02 ± 3.12 N) were below the specification limit (39.23 N, USP 2019). The weight variation results of all generic products complied with the USP specification requirement. In the quantitative assay results, the minimum and maximum API contents were 97.4 ± 0.02 (NF6) and 105.8 ± 0.02 (NF8), respectively, which are within the limit specified by the USP (90%–110%). Similarly, all samples met the disintegration time limit (i.e., ≤30 min) and drug-releasing tolerance limit (API released more than 60% within 60 min) requirements. The f2 values were >50, and the f1 values were <15 for all sampled brands.ConclusionThe majority of the sampled brands of HCT tablets met the quality requirements as per USP official test specifications. From the similarity and difference factor values, all studied brands were shown to be equivalent.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2024.1421463</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2024.1421463</link>
        <title><![CDATA[Separation of isobaric phosphorothioate oligonucleotides in capillary electrophoresis: study of the influence of cationic cyclodextrins on chemo and stereoselectivity]]></title>
        <pubdate>2024-06-13T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Maryam K. Ghassemi</author><author>Vincent Hurlet</author><author>Jacques Crommen</author><author>Anne-Catherine Servais</author><author>Marianne Fillet</author>
        <description><![CDATA[Phosphorothioate (PS) oligonucleotides have drawn more and more attention lately due to their great therapeutic potential. The presence of one (or several) phosphorothioate moiety (ies) improves pharmacokinetic properties but at the same time creates an additional chiral center for each phosphorothioate linkage, and thus diastereomers. It is therefore important to develop analytical strategies to monitor individual species to enable more in-depth investigations. In this study, a PVA coated capillary with a background electrolyte made of 100 mM phosphoric acid adjusted to pH 3.0 with triethanolamine was used. A design of experimental approach provides the optimal conditions for the separation of the eight isobaric diastereomers bearing one phosphorothioate linkage (Mix 1), and the separation of the 12 isobaric diastereomers of a mixture made of oligonucleotides with two phosphorothioate linkages (Mix 2). Remarkably, full separation in Mix 1 could be achieved using a combination of a cationic cyclodextrin (2-hydroxy-3-N,N,N-trimethylamino) propyl-γ-CD chloride, and an anionic cyclodextrin (carboxymethyl-β-cyclodextrin sodium salt), while a second cationic cyclodextrin (2-hydroxy-3-N,N,N-trimethylamino) propyl-β-CD chloride) was required for Mix 2, providing additional selectivity.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2024.1358893</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2024.1358893</link>
        <title><![CDATA[Reliable particle sizing in vaccine formulations using advanced dynamic light scattering]]></title>
        <pubdate>2024-02-29T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Coline Bretz</author><author>Andrea Jauslin</author><author>Dario Leumann</author><author>Marius Koch</author><author>Andrea Vaccaro</author>
        <description><![CDATA[Understanding the impact of lipid nanoparticle size on immunogenicity represents an important step for enabling the rapid development of novel vaccines against known or emergent diseases. Dynamic light scattering, also known as quasi-elastic light scattering or photon correlation spectroscopy, has established itself as an optimal analytical method to determine particle size due to its in-situ approach and fast measurements. However, its application to many systems of industrial relevance has been hindered due to artifacts arising from multiple scattering. Result interpretation becomes severely compromised depending on the concentration of the system and the size of the particles. In this context, strong sample dilution is often required, bringing additional uncertainties to the formulation development process. Here, we show how advanced dynamic light scattering technology can filter out multiple scattering from the signal and yield fully accurate sizing measurements regardless of the sample concentration. We illustrate this in a comparative study with standard dynamic light scattering using polystyrene beads as model suspension as well as a concentrated commercial lipid nanoparticle adjuvant (AddaVax™).]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2023.1154391</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2023.1154391</link>
        <title><![CDATA[Heparin increases the antibiotic efficacy of colistin]]></title>
        <pubdate>2023-05-12T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Gergo Peter Szekeres</author><author>Emeline Hanozin</author><author>Robyn Diehn</author><author>Jan Horlebein</author><author>Lukasz Polewski</author><author>Andreas Zappe</author><author>Daniel Lauster</author><author>Kevin Pagel</author>
        <description><![CDATA[The increasing antibiotic resistance in bacteria is an alarming phenomenon all around the world. Certain strains have developed resistance against multiple antimicrobial molecules, in which cases, the final option is to use a last-resort drug. These drugs, however, are last-resort for a reason: they can pose serious risk on vital organ functions in the patient. To mitigate the risk of severe side-effects and to reduce the rate of bacterial mutation, co-administration with other molecules that increase their efficacy seems to be the only suitable option. This leads to a reduced dose while maintaining the same level of antibiotic activity within the body. In this study, the effect of heparin derivatives on the antibiotic activity of colistin and their interactions were studied by ion mobility, mass spectrometry, and bacterium growth assays. The results show that during the association of colistin and heparin, they retain their structure while higher-stoichiometry complexes can form. When long-chain heparin is co-administered, multiple colistin molecules can associate with it, which increases the antibiotic activity by ∼40% relative to the sole administration of colistin.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2023.1175290</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2023.1175290</link>
        <title><![CDATA[Editorial: The protagonism of bioanalytical methods in high-throughput drug discovery]]></title>
        <pubdate>2023-04-11T00:00:00Z</pubdate>
        <category>Editorial</category>
        <author>Marcela Cristina de Moraes</author><author>Fernando Gonçalves de Almeida</author><author>Luzineide Wanderley Tinoco</author>
        <description></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2023.1091764</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2023.1091764</link>
        <title><![CDATA[Investigating metformin-active substances from different manufacturing sources by NIR, NMR, high-resolution LC-MS, and chemometric analysis for the prospective classification of legal medicines]]></title>
        <pubdate>2023-03-07T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Mariangela Raimondo</author><author>Francesca Prestinaci</author><author>Federica Aureli</author><author>Giulia D’Ettorre</author><author>Maria Cristina Gaudiano</author>
        <description><![CDATA[Introduction: The characterisation of active substances is an essential tool to ensure the traceability and authenticity of legal medicines. Metformin is a well-established biguanide derivative recommended in oral formulations as a first-line treatment for type 2 diabetes. With its increasing demand, metformin is likely to be an attractive target for falsification and substandard production, thus posing health risks to consumers. Methods that are able to identify even small differences in active pharmaceutical ingredients (APIs) are deemed necessary. The detection of fraudulent practices in APIs is not straightforward, and a single technique that can provide sufficient information to unambiguously address this issue is still not available.Methods: This study investigated an integrated analytical platform based on NIR, 1H-NMR, 13C-NMR, and high-resolution LC-MS combined with chemometrics to profile 32 metformin hydrochloride samples originating from several global authorised manufacturers. The study's aim was to explore differences in the chemical characteristics of metformin hydrochloride APIs to identify or predict a possible classification for each manufacturer in view of prospective authenticity studies. Different pre-processing methods were applied; bucket tables for 1H- and 13C-NMR were obtained, while mass spectrometry data were processed in targeted and untargeted modes. Datasets were individually analysed and merged by a multivariate unsupervised method and performing principal component analysis (PCA). Results and Discussion: The results evidenced differences in cluster behaviour, depending on manufacturers. Each technique has shown a specific clustering tendency, highlighting how different analytical approaches are able to characterise metformin APIs. Some manufacturers’ samples, however, showed similar behaviour independently of the techniques. NIR and 1H-NMR were confirmed as the more predictive techniques if taken individually; 1H-NMR, in particular, achieved good separation between the samples of the two most representative manufacturers. For LC-MS, the targeted approach resulted in a separation in groups clearer than that of the untargeted approach. Nevertheless, the untargeted LC-MS approaches presented in this paper could be a possible alternative to obtaining different information for drug substances, with several different and complex synthetic pathways leading to several unknown impurities. Further grouping of manufacturers emerged by data fusion, highlighting its potential in the traceability of metformin.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2023.1059884</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2023.1059884</link>
        <title><![CDATA[Advances and strategies for capillary electrophoresis in the characterization of traditional Chinese medicine: A review of the past decade (2011–2021)]]></title>
        <pubdate>2023-02-22T00:00:00Z</pubdate>
        <category>Review</category>
        <author>Shahab A. Shamsi</author><author>Jalpaben Patel</author>
        <description><![CDATA[While traditional Chinese medicine (TCM) is considered a valuable resource for drug discovery and form a potential basis for drug development, they also carry substantial safety risks due to adverse drug reactions and a lack of understanding of their mechanisms of action. However, due to their highly complex composition, valid analytical methodologies for analyzing TCMs must be developed and promoted. An extensive search of published research and review of scientific papers implies that the increased efficiency and sensitivity of capillary electrophoresis (CE) has attracted much research attention. This review provides an in-depth assessment of CE applications for TCM analysis published in the open literature in the last decade (2011–2021). Our survey findings showed that capillary zone electrophoresis (CZE) with ultraviolet (UV) detection is a capillary electromigration technique frequently utilized for the efficient separation, identification, and quantitation of various active components in highly complex matrices. Different extraction methods, modifiers to the background electrolyte, preconcentration techniques, and mass spectrometry (MS) detectors are used to enhance CE separation selectivity and TCM sensitivity.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2023.1066348</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2023.1066348</link>
        <title><![CDATA[Development and validation of an ultraviolet-visible spectrophotometric method for quantifying oxytetracycline for veterinary use]]></title>
        <pubdate>2023-02-22T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Jacques Mbuyi Kaluka Tshibamba</author><author>Jocelyn Mankulu Kakumba</author><author>Timothy Mavanga Mabaya</author><author>Roland Marini Djang’ieng’a</author><author>Jérémie Mbinze Kindenge</author>
        <description><![CDATA[Livestock breeding plays a key role in sub-Saharan Africa because it is an important source of highly valued protein in the human diet, and because it is an essential driver for socio-economic development. It represents a form of investment and is also important from a socio-cultural point of view (ceremonies, social position, etc.). Livestock is an important source of income, livelihood, nutrition, and food security. However, livestock breeding faces, among other things, major sanitary constraints. Furthermore, the circulation of non-compliant antibiotics on the market constitutes a major threat to animal health, public health, and the environment. This research aims to develop and validate a UV-vis method for quantifying pharmaceutical oxytetracycline. The method developed was validated following the total error strategy as a decision tool in the accuracy profile. After its completion, the method demonstrated good absolute and relative bias and was within a tolerable interval of [−2%, +2%]. The method was also repeatable with intermediate precision, with respectively lower values than 2% and 4%. We also assessed the recovery and accuracy of the method as fitting with the specification limits. After its validation, the method was quantified in 47 oxytetracycline injectable samples, where we obtained 28 samples complying with specifications and 19 that did not. That led us to conclude that the developed method was validated and appropriate for quantification in terms of the routine quality control of oxytetracycline injection. The method needs to be reviewed and revalidated accordingly for other pharmaceutical presentations.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2022.1091206</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2022.1091206</link>
        <title><![CDATA[Direct analysis of tobacco specific nitrosamines in tobacco products using a molecularly imprinted polymer-packed column]]></title>
        <pubdate>2023-01-06T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Haley A. Mulder</author><author>Justin L. Poklis</author><author>Adam C. Pearcy</author><author>Matthew S. Halquist</author>
        <description><![CDATA[Tobacco specific nitrosamines (TSNAs) are highly carcinogenic by-products in tobacco samples, and their presence is regulated by the Food and Drug Administration. Molecularly imprinted polymers (MIPs) are synthetic polymers that have been “imprinted” with a template analyte in a co-polymer system, and can selectively extract analytes from complex matrices. MIPs can be incorporated into online systems, replacing traditional high performance liquid chromatography (HPLC) columns. MIP material specific for TSNAs was packed into an empty HPLC column using a slurry packing technique. The developed method with the MIP-packed HPLC column was validated on a LC-MS/MS system for the quantitation of N-nitrosonornicotine (NNN) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in commercial tobacco products. The method was linear over .1–10 ng/ml (.4–10 μg/g) for NNN and NNK. The limit of detection (LOD) was .03 ng/ml (12 μg/g) and the limit of quantitation (LOQ), .1 ng/ml (.4 μg/g). All column uniformity parameters with the exception of theoretical plate number were within the accepted criteria (%RSD values <15%). Theoretical plate number was <250, owing to the large (50 μm) sized MIP particles. Twenty-six tobacco products contained TSNA concentrations that were consistent with reported literature values. The TSNA-MIP based HPLC column effectively replaced a traditional reverse phase HPLC column, and was used for the direct analysis of nicotine and tobacco products without extensive sample preparation prior to instrumental analysis.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2022.1028752</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2022.1028752</link>
        <title><![CDATA[Sterilization of micronized indomethacin]]></title>
        <pubdate>2023-01-06T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Karyna Krupianskaya</author><author>Bart Croonenborghs</author><author>Annick Gillet</author><author>Bernard Appeltans</author><author>Anke Meyers</author><author>Aaron DeMent</author><author>Guy Van den Mooter</author><author>Ann Van Schepdael</author><author>Erik Haghedooren</author>
        <description><![CDATA[Sterilization is a pivotal topic in the pharmaceutical industry, whereby the nomenclature of “sterile” refers to the absence of viable microorganisms. Since microorganisms can reproduce in the body and cause potentially fatal infections, it is critical to sterilize parenteral products to prevent this. In recent years, 70%–90% of potential drugs and 40% of marketed drugs have demonstrated a low solubility. Micronization is a widely spread approach to increase the dissolution rate. A subset of micronized products require sterilization, but published studies on the effects of sterilization on micronized products are currently lacking. The effect of sterilization on the micronized active pharmaceutical ingredient indomethacin was explored in this study. The sterilization methods in scope were one photon-based method using gamma irradiation and one gas-based method with nitrogen dioxide gas. Indomethacin was micronized using two micronization techniques, cryomilling and spray drying. Different conditions were used for cryomilling where the number of grinding balls in the ball mill and the degree of filling were varied. The solid state of all samples was evaluated after micronization, and only the effectively micronized samples were selected for sterilization with gamma rays and nitrogen dioxide. Gamma irradiation was performed with the active pharmaceutical ingredient stored at −80°C at a commonly used industry standard target dose of 25 kGy. Nitrogen dioxide sterilization took place at 21°C, a concentration of 10 mg/L, a relative humidity of 30% and using two NO2 pulses. Before and after sterilization, all samples were analyzed by high performance liquid chromatography with UV detection, whereby the assay of indomethacin was examined as well as the peak purity and the formation of impurities. In comparison to the non-micronized reference, both sterilization methods demonstrate a significant decrease of content of micronized samples and an increase of the impurity profile. The non-micronized sample showed no significant difference after sterilization. It could be observed that micronized indomethacin samples demonstrate more degradation and are subsequently more susceptible to degradation upon sterilization with gamma rays and nitrogen dioxide gas, driving towards the need for assessment of the micronization impact combined with sterilization approach.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2022.1018115</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2022.1018115</link>
        <title><![CDATA[Activity assay based on the immobilized enzyme kallikrein and mass spectrometry]]></title>
        <pubdate>2022-10-19T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Daniella Romano De Carvalho</author><author>Bruna Barbosa Laurentino</author><author>Camila Loreta Rocha</author><author>Jeroen Kool</author><author>Govert Somsen</author><author>Erika Amstalden van Hove</author><author>Carmen Lúcia Cardoso</author>
        <description><![CDATA[Deregulated activity and expression of human kallikreins (KLKs) may be involved in various pathologies, so these enzymes are an attractive biological target for identifying molecules that can modulate KLK activity. This identification involves applying fast and efficient screening methods. This work describes an off-line assay with mass spectrometry (MS) detection that uses KLK immobilized on Sepharose-NHS as a micro-column configuration (IMER-KLK-Sepharose-NHS). The mass spectrometry used has an ion trap analyzer and electrospray ionization (EIS). The HPLC-MS method for quantifying KLK activity was developed. The enzymatic assay conditions were optimized, and the IMER-KLK-Sepharose-NHS kinetic parameter (KMapp = 15.48 ± 3 μmol L−1) was evaluated. Finally, the method was validated by using leupeptin as a reference inhibitor (IC50 = 0.85 ± 0.10 μmol L−1). The developed method was able to identify the reference inhibitor and can be an alternative for screening KLK inhibitors.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2022.1018080</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2022.1018080</link>
        <title><![CDATA[Analytical assays to evaluate enzymatic activity and screening of inhibitors for ornithine decarboxylase]]></title>
        <pubdate>2022-10-10T00:00:00Z</pubdate>
        <category>Mini Review</category>
        <author>Luzineide W. Tinoco</author><author>Bruno Maia da Silva Santos</author><author>Jhones Matheus da Silva Soares</author><author>Fernanda Gadini Finelli</author>
        <description><![CDATA[Ornithine decarboxylase (ODC) catalyzes the decarboxylation of ornithine to produce putrescine, the first step in the metabolism of polyamines (putrescine, spermidine, and spermine), which are essential growth factors in eukaryotic cells. ODC is active as a homodimer and depends on pyridoxal 5′-phosphate (PLP) as a cofactor. An increase in the concentration of polyamines has been associated with carcinogenesis. Therefore, there is much interest in identifying inhibitors of this pathway as potential chemotherapeutic and chemopreventive agents. The best-known inhibitor of mammalian ODC is α-difluoromethylornithine (DFMO), a highly selective compound that alkylates Cys-360 (a residue of the ODC active site). Although DFMO was initially developed for the treatment of cancer, the World Health Organization recommends its use in combination with nifurtimox for the treatment of human African trypanosomiasis. Considering the importance of ODC as a promising target for the treatment of various types of cancer and other infectious diseases, choosing the right method for screening potential inhibitors can help to accelerate the discovery of new drugs. Several methods for the determination of ODC activity are found in the literature. Among these, we can mention analysis with radioactive markers, colorimetric assays using auxiliary enzymes to detect CO2 or H2O2 release, chromatographic separations with putrescine derivatization, mass spectrometry, and spectroscopic techniques. In this review, the main analysis methods used will be described, highlighting their advantages and disadvantages, as well as identifying the most promising methods for high-throughput screening.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2022.1002935</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2022.1002935</link>
        <title><![CDATA[Making high salt concentrations for optimal chromatography compatible with electrospray ionization mass spectrometry using an ion exchange membrane suppressor: Analysis of biomarkers for transporter protein inhibition as a case study]]></title>
        <pubdate>2022-09-12T00:00:00Z</pubdate>
        <category>Original Research</category>
        <author>Sam Wouters</author><author>Ils Pijpers</author><author>Ninon Vanden Haute</author><author>Daniel Meston</author><author>Lieve Dillen</author><author>Filip Cuyckens</author><author>Sebastiaan Eeltink</author>
        <description><![CDATA[This study reports on the potential of using ion-exchange suppressor technology in liquid chromatography—electrospray ionization mass spectrometry workflows. The aim was to use high salt concentrations to improve separation performance, while overcoming the resulting significant ion suppression during electrospray ionization. As a case study, we apply suppressor technology to the hydrophilic interaction liquid chromatography separation and detection of taurine and glycochenodeoxycholate sulfate, endogenous biomarkers for organic anion transporter protein inhibition. The desired chromatographic selectivity was achieved applying 100 mM ion-pairing agent, while competing ions negatively affecting MS sensitivity were actively removed post-column from the solvent via a charged partially permeable membrane and replaced with protons, resulting in an up to 10-fold increase in detection sensitivity.]]></description>
      </item><item>
        <guid isPermaLink="true">https://www.frontiersin.org/articles/10.3389/frans.2022.1004113</guid>
        <link>https://www.frontiersin.org/articles/10.3389/frans.2022.1004113</link>
        <title><![CDATA[On-flow enzymatic inhibitor screening: The emerging success of liquid chromatography-based assays]]></title>
        <pubdate>2022-09-07T00:00:00Z</pubdate>
        <category>Review</category>
        <author>Pamella Christina Ortega De Oliveira</author><author>Renato Côrrea Lessa</author><author>Millena Santana Ceroullo</author><author>Camila Anchau Wegermann</author><author>Marcela Cristina De Moraes</author>
        <description><![CDATA[Enzymes are targets commonly explored in screening assays aiming to discover new leads in the drug development process. Among the diverse assay models to identify new enzymatic inhibitors, on-flow assays based on liquid chromatography (LC) can be highlighted. In these approaches, the ligand-enzyme interaction can be examined by monitoring the catalytic activity or the affinity/retention. Most applications use the biological target immobilized in solid supports resulting in the acquisition of an immobilized enzymatic reactor (IMER). Coupling IMERs to LC or mass spectrometry (MS) systems allows monitoring enzyme activity online and studying binding events between target and ligands. On-flow screening assays present many advantages for the hit-to-lead process, such as the possibility of system automation, reusability, and high stability. This review covers articles from the last decade that combine the use of varied immobilization methods on different solid supports and several equipment setups in on-flow systems, emphasizing the performance and capacity of recognizing and identifying biologically active compounds in various matrices.]]></description>
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