{"title":"The acute oral toxicity and pharmacokinetic determination of a novel chloroquinoline hybrid molecule by LC-MS/MS.","authors":"Monique Labuschagne, Makhotso Lekhooa, Thrineshen Moodley, Chakes Mashaba, Matshawandile Tukulula","doi":"10.1080/17576180.2025.2567227","DOIUrl":"https://doi.org/10.1080/17576180.2025.2567227","url":null,"abstract":"<p><strong>Background: </strong>CB-1 is a novel compound evaluated in Sprague Dawley rats to assess its acute oral safety and pharmacokinetic disposition.</p><p><strong>Objectives: </strong>To determine the short-term oral toxicity of CB-1 and characterize its pharmacokinetic parameters.</p><p><strong>Methods: </strong>Acute oral toxicity was tested at doses up to 100 mg/kg. For pharmacokinetics, six rats received a single 100 mg/kg oral dose. Plasma samples were analyzed using a polar C18 column and quantified by LC - MS/MS monitoring the 490.063→262.00 m/z transition.</p><p><strong>Results: </strong>No mortality occurred at 100 mg/kg, indicating acceptable acute safety. CB-1 reached a peak plasma concentration (C<sub>max</sub>) of 4324.24 ng/mL at 1 h (T<sub>max1</sub>). Pharmacokinetic analysis revealed an apparent clearance (Cl/F) of 3.6 L/h, elimination half-life (t<sub>1/2</sub>) of 7.86 h, and volume of distribution (Vd/F) of 40 L. A secondary plasma concentration peak at 8 h (T<sub>max2</sub>) suggested enterohepatic recirculation.</p><p><strong>Conclusions: </strong>CB-1 demonstrated short-term oral safety with rapid absorption, relatively low clearance, and prolonged systemic exposure, likely influenced by enterohepatic recirculation.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-8"},"PeriodicalIF":1.8,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145197857","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioanalysisPub Date : 2025-09-29DOI: 10.1080/17576180.2025.2565142
Shivam Rathaur, Parag Varshney, Sachin Vishwakarma, Debalina Maity, Sharib R Khan, Jiaur R Gayen
{"title":"Simultaneous estimation of finerenone and canagliflozin using HPLC: application in metabolic stability studies.","authors":"Shivam Rathaur, Parag Varshney, Sachin Vishwakarma, Debalina Maity, Sharib R Khan, Jiaur R Gayen","doi":"10.1080/17576180.2025.2565142","DOIUrl":"https://doi.org/10.1080/17576180.2025.2565142","url":null,"abstract":"<p><strong>Aim: </strong>A robust, sensitive, and reliable method was developed and validated on HPLC for the simultaneous estimation of Finerenone (FNR) and Canagliflozin (CFZ).</p><p><strong>Method: </strong>The resolution was performed by using mobile-phase acetonitrile (ACN): Water (51:49) at a flow rate of 0.75 ml/min with the C18 analytical column Phenomenex Luna (250 mm, 4.6 mm, 5 µm). FNR and CFZ were estimated at a retention time of 6.33 and 8.26 min with 10.0 min analysis run time and detected by PDA detector at a λ<sub>max</sub> 249 and 290 nm, respectively. The calibration curve was linear over the concentration range of 0.05-10 µg/ml with R<sup>2</sup> 0.998 and 0.999 for FNR and CFZ, respectively. For FNR and CFZ, the limit of detection (LOD) was 0.53 and 0.36 μg/ml & limit of quantitation (LOQ) was 1.62 and 1.10 μg/ml, respectively. The method was validated using specificity, linearity, accuracy, precision, LOD, LOQ, robustness, and stability.</p><p><strong>Conclusion: </strong>According to the International Council for Harmonisation (ICH) guideline, the validation studies confirmed that the optimization method is specific, simple, highly sensitive, reliable, robust, and reproducible. The developed method was successfully applied for simultaneous estimation with applications in in-vitro metabolic stability studies of pooled microsomes of humans, monkeys, dogs, rabbits, rats and mice.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-14"},"PeriodicalIF":1.8,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145184645","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioanalysisPub Date : 2025-09-29DOI: 10.1080/17576180.2025.2567231
Osama I Abdel Sattar, Hamed H M Abuseada, Mohamed S Emara, Islam Selim, Nahla A Abdelshafi
{"title":"A clever evaluation of the antidiabetic medications linagliptin and empagliflozin in bulk and spiked urine samples.","authors":"Osama I Abdel Sattar, Hamed H M Abuseada, Mohamed S Emara, Islam Selim, Nahla A Abdelshafi","doi":"10.1080/17576180.2025.2567231","DOIUrl":"https://doi.org/10.1080/17576180.2025.2567231","url":null,"abstract":"<p><strong>Background: </strong>The objective of this research is to provide differential pulse voltammetry (DPV) method that is an ingenious, simple, sensitive, and selective method for the quantitative analysis of linagliptin and empagliflozin in spiked human urine samples. Faster analysis times and the use of small sample volumes are great advantages of DPV.</p><p><strong>Research design and methods: </strong>Before centrifuging the samples for 5 min at 3000 rpm, methanol was added to precipitate and remove any suspended compounds. As a working electrode, glassy carbon electrode (GCE) was employed. The auxiliary electrode was a platinum electrode, while the reference electrode was Ag|AgCl|KCl(sat.). To maximize the experimental conditions for simultaneous determination, the relationship between the current, pH and scan rate was examined.</p><p><strong>Results: </strong>Optimal conditions for quantitative determination were obtained in a phosphate buffer (PB) at pH 7. Plotting LIN and EMP concentrations against the DPV peak height revealed good linearity. It was found that the linear ranges for LIN and EMP were 10-90 and 10-70 µM, respectively.</p><p><strong>Conclusions: </strong>This approach has been designed for effectively estimating the levels of examined drugs in human urine without matrix effect. The obtained results showed a good recovery values of both drugs.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-8"},"PeriodicalIF":1.8,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145190685","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioanalysisPub Date : 2025-09-26DOI: 10.1080/17576180.2025.2565141
Tímea Dérerová, Zuzana Vosáhlová, Květa Kalíková
{"title":"Recent advances in analytical separation techniques for therapeutic oligonucleotides.","authors":"Tímea Dérerová, Zuzana Vosáhlová, Květa Kalíková","doi":"10.1080/17576180.2025.2565141","DOIUrl":"https://doi.org/10.1080/17576180.2025.2565141","url":null,"abstract":"<p><p>Therapeutic oligonucleotides are an emerging class of drugs designed for gene expression modulation. The increasing number of clinical trials and currently expanding market is facilitating further integration and accessibility of these therapeutics. A crucial step in drug development involves reliable analytical tools for characterization and quality control. For clinical applications, oligonucleotides must be separated and purified to ensure regulatory compliance. However, their analysis represents a complex bioanalytical challenge, grounded in their complex impurity profiles. Chemical stability and binding affinity of oligonucleotide-based therapeutics are enhanced during synthesis by extensive modifications, inducing formation of various synthesis failures or truncated sequences. Furthermore, meeting current guidelines or addressing manufacturing scale-up strategies remains challenging as each oligonucleotide typically necessitates a custom analytical protocol. Here, we provide an overview of the most recent advances in separation methods, including various chromatography methods and capillary electrophoresis for nucleic acid-based therapeutics.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-16"},"PeriodicalIF":1.8,"publicationDate":"2025-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145172835","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioanalysisPub Date : 2025-09-22DOI: 10.1080/17576180.2025.2554565
Abdullah Zia, Toshifumi Yokota
{"title":"Upgrading nucleic acid and antisense therapeutics: challenges, solutions, and future directions.","authors":"Abdullah Zia, Toshifumi Yokota","doi":"10.1080/17576180.2025.2554565","DOIUrl":"https://doi.org/10.1080/17576180.2025.2554565","url":null,"abstract":"<p><p>Only a small fraction of disease-modifying proteins present druggable pockets for conventional small-molecule or biologic therapies, underscoring the urgent need for innovative strategies such as nucleic acid-based antisense therapeutics. Antisense approaches-including antisense oligonucleotides (ASOs), RNA interference (RNAi), and decoy oligodeoxynucleotides (ODNs)-offer powerful means to directly modulate gene expression at the RNA level. Over the past four decades, these modalities have advanced from early proof-of-concept studies to numerous FDA- and EMA-approved therapies for neuromuscular, metabolic, and neurodegenerative diseases. Despite these successes, critical barriers remain. Antisense drugs face challenges related to nuclease degradation, off-target binding, dose-dependent toxicities, limited tissue penetration, and inefficient endosomal escape. Addressing these limitations will require advances in nucleotide chemistry, conjugation strategies, and delivery platforms. Personalized \"N-of-1\" therapies further highlight the promise of customized oligonucleotides but also raise ethical and cost considerations. This review synthesizes the current state of antisense modalities, the obstacles impeding their broader application, and the innovative approaches needed to upgrade existing platforms and expand their therapeutic potential across a wider range of genetic and acquired diseases.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-9"},"PeriodicalIF":1.8,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145111854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioanalysisPub Date : 2025-09-14DOI: 10.1080/17576180.2025.2557182
Arisa Araki, Mahiro Takano, Christian Nanga Chick, Takumi Yamazaki, Jun Nojima, Toyonobu Usuki
{"title":"Correlation between LC-MS/MS and ELISA methods for quantitative analysis of desmosine-containing solutions.","authors":"Arisa Araki, Mahiro Takano, Christian Nanga Chick, Takumi Yamazaki, Jun Nojima, Toyonobu Usuki","doi":"10.1080/17576180.2025.2557182","DOIUrl":"https://doi.org/10.1080/17576180.2025.2557182","url":null,"abstract":"<p><strong>Background: </strong>Desmosine and isodesmosine are crosslinking amino acids found in extracellular matrix protein elastin, which imparts elasticity to tissues such as those of the lungs and arteries. These compounds are promising biomarkers for diseases involving elastin degradation, such as chronic obstructive pulmonary disease.</p><p><strong>Research design and method: </strong>This study examined the correlation between isotope-dilution LC-MS/MS and a newly established ELISA for in vitro diagnosis using a variety of samples.</p><p><strong>Results: </strong>Results of ELISA and LC-MS/MS analyses exhibited a high correlation coefficient (0.9941). However, whereas the LC-MS/MS measurements deviated approximately 2-fold from the theoretical values, the ELISA measurements ranged from 0.83 to 1.06 (avg 0.94) times the theoretical values. Precise measurement of the absorbance of synthetic desmosine revealed a molar extinction coefficient of 2403, which differed markedly from the previously reported value of 4900 in 1963. Using this value to recalculate the amount of added desmosine, the LC-MS/MS measurements were 0.68 to 0.99 (avg 0.87) times the theoretical values.</p><p><strong>Conclusion: </strong>Thus, the developed ELISA enables highly accurate determination of desmosine concentrations, comparable to LC-MS/MS, suggesting that ELISA is a potentially useful in vitro diagnostic tool.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-5"},"PeriodicalIF":1.8,"publicationDate":"2025-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145063418","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioanalysisPub Date : 2025-09-07DOI: 10.1080/17576180.2025.2555774
Philip Timmerman, Stuart McDougall, Neil Adcock, Cecilia Arfvidsson, Matthew Barfield, Stefan Blech, Kyra J Cowan, Luca Ferrari, Alessandro Greco, Michaela Golob, Lee Goodwin, Richard Hughes, Tsvetelina Ivanova, Anna Laurén, Robert Nelson, Sonja Neitzel, Tom Verhaeghe, Nico van de Merbel, Mike Wright, Stephen White
{"title":"European Bioanalysis Forum recommendation on embracing a context-of-use-driven scientific validation for chromatographic assays in the light of ICH M10.","authors":"Philip Timmerman, Stuart McDougall, Neil Adcock, Cecilia Arfvidsson, Matthew Barfield, Stefan Blech, Kyra J Cowan, Luca Ferrari, Alessandro Greco, Michaela Golob, Lee Goodwin, Richard Hughes, Tsvetelina Ivanova, Anna Laurén, Robert Nelson, Sonja Neitzel, Tom Verhaeghe, Nico van de Merbel, Mike Wright, Stephen White","doi":"10.1080/17576180.2025.2555774","DOIUrl":"https://doi.org/10.1080/17576180.2025.2555774","url":null,"abstract":"<p><p>The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use M10 guideline provides a global framework for bioanalytical method validation in studies intended for regulatory submission. While its structure ensures consistency and data reliability, the guideline also acknowledges that not all studies require the same level of validation. This paper examines where full compliance is essential and where scientific judgment allows for leaner, context-driven validation, such as in early-stage development, for additional matrices, metabolites, nonstandard biological matrices or studies intended for internal decision-making. Drawing on recent recommendations from the European Bioanalysis Forum, we introduce a decision-making flowchart and parameter table to support consistent application of a Context-of-Use approach to validation. These tools help guide when flexibility is appropriate while ensuring transparency and robustness in the data. The paper advocates continued dialogue both with end users of the data and the regulatory authorities to support a modernized, risk-based validation framework that remains aligned with patient needs and scientific integrity. We believe the recommendations in this paper are fully in alignment with the intent and core principles of ICH M10, while encouraging their application in a way that remains scientifically driven and proportionate to the purpose of the assay.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-10"},"PeriodicalIF":1.8,"publicationDate":"2025-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145013792","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BioanalysisPub Date : 2025-09-05DOI: 10.1080/17576180.2025.2554568
Ahmar Khan, Claire Clinton, Jennifer Hawkins, John Robinson, Sam Maynard, Shiva Jalili, Tarin Taleb, Jack Lodge
{"title":"An interview with <i>Bioanalysis</i>: speaking with the 2025 international reid bioanalytical forum bursary award winners.","authors":"Ahmar Khan, Claire Clinton, Jennifer Hawkins, John Robinson, Sam Maynard, Shiva Jalili, Tarin Taleb, Jack Lodge","doi":"10.1080/17576180.2025.2554568","DOIUrl":"10.1080/17576180.2025.2554568","url":null,"abstract":"","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-5"},"PeriodicalIF":1.8,"publicationDate":"2025-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144999495","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}