Journal of pharmaceutical and biomedical analysis最新文献

{"title":"Unraveling the cytotoxicity and cellular pharmacokinetic of mPEG5-NH2 polymers by UHPLC-MS/MS","authors":"Yingxia Guo ,&nbsp;Meichen Liu ,&nbsp;Yajie Cheng ,&nbsp;Jiye Tian ,&nbsp;Chunpeng Feng ,&nbsp;Qingbin Wang ,&nbsp;Xuan Zhao ,&nbsp;Lei Yin","doi":"10.1016/j.jpba.2025.116767","DOIUrl":"10.1016/j.jpba.2025.116767","url":null,"abstract":"<div><div>Methoxy-polyethylene glycol amine (mPEG-NH₂), an important pharmaceutical excipient, has been extensively utilized in the field of biomedicine. To advance the development of mPEG-NH₂-related drug delivery systems, it is crucial to understand the safety profile and cellular pharmacokinetic behavior of mPEG-NH₂ polymers. In this study, a straightforward analytical assay based on ultra-high performance liquid chromatography and tandem mass spectrometry (UHPLC-MS/MS) for quantifying mPEG₅-NH₂ in biological matrices was established and validated. Multiple reaction monitoring (MRM) transitions were selected for quantification of mPEG₅-NH₂ (mass-to-charge ratio, <em>m/z</em> 252.1 → 87.7) and octaethylene glycol (OH-PEG₈-OH) (mass-to-charge ratio, <em>m/z</em> 371.2 → 89.2). The UHPLC-MS/MS assay demonstrated excellent linearity within the concentration range of 0.01–10 μg/mL, with an R² value of 0.9996. Both intra-day and inter-day accuracies and precisions of the analytical method were within ± 9.19 %. This analytical assay was successfully applied to study the <em>in vitro</em> cellular toxicity and uptake behavior of mPEG₅-NH₂ in MCF-7 cells. The results indicate that high doses of mPEG₅-NH₂ may have potential toxicity to MCF-7 cells.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116767"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143521016","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of oral fluid in urine samples on the analysis of selected erythropoietin receptor agonists and detection of saliva-specific markers for doping control purposes","authors":"Ann-Marie Garzinsky ,&nbsp;Judith Harth ,&nbsp;Florine Leipp ,&nbsp;Katja Walpurgis ,&nbsp;Philipp Reihlen ,&nbsp;Andreas Thomas ,&nbsp;Mario Thevis","doi":"10.1016/j.jpba.2025.116769","DOIUrl":"10.1016/j.jpba.2025.116769","url":null,"abstract":"<div><div>Due to their performance-enhancing effect, erythropoiesis-stimulating agents (ESAs) are banned at all times by the World Anti-Doping Agency (WADA) in competitive sports. Doping control analyses for such compounds are routinely performed using gel electrophoretic and immunoblotting techniques, and degradation of the analytes can severely impair detection, results evaluation and interpretation. As oral fluid (OF) contains significant amounts of proteases, the question of whether its addition to a doping control urine sample may impede anti-doping analysis needs to be addressed. Intentional tampering attempts are likewise prohibited by WADA and require a detection strategy. It was observed that the addition of OF can indeed lead to impairments of ESA analyses, though the fraction of unidentifiable ESA signals varies depending on several factors, such as the individual composition of the OF, the sex of the OF donor, the time of sampling, the OF volume and the incubation conditions. Overall, 20 % of all generally valid analyses were classified as unidentifiable, 12 % as impaired, and 69 % as identifiable, highlighting the relevance for strategies that allow for the identification of OF in urine. While human salivary α-amylase was found insufficiently reliable as a marker, peptides of salivary proline rich proteins (saPRP) were shown to be both specific for OF and traceable with adequate sensitivity using a newly developed LC-HRMS/MS method. The approach was comprehensively characterized shown to be fit-for-purpose for routine doping controls where tampering attempts with OF are suspected.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116769"},"PeriodicalIF":3.1,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143509678","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of chemical components and metabolites in rats after oral administration of Epimedium-Astragalus granule pair by liquid chromatography-high resolution mass spectrometry combined with diagnostic fragment ions and mass defect filtering","authors":"Song Wang ,&nbsp;Xinnan Chang ,&nbsp;Jing Li ,&nbsp;Zuoqiao Shi ,&nbsp;Guowen Li","doi":"10.1016/j.jpba.2025.116768","DOIUrl":"10.1016/j.jpba.2025.116768","url":null,"abstract":"<div><div>Herbal pairs are combinations of two relatively fixed herbs that are frequently used in clinical practice to achieve specific therapeutic effect. Epimedium and Astragalus are frequently used together in clinical settings. However, there is a lack of an in-depth understanding of the active components of these herbs <em>in vivo</em>. In this study, a method based on ultra-high-performance liquid chromatography coupled to high resolution mass spectrometry together with diagnostic fragment ions (DFIs) mass defect filtering (MDF) was developed to systematically screen and identify the chemical ingredients presenting in Epimedium-Astragalus granule pair (EAGP) and the absorbed components and their metabolites in rat plasma following oral administration. Using accurate mass determination, mass defect filtering and diagnostic fragment ion screening strategies, a total of eighty-five ingredients were identified in EAGP. By comparing the total ion chromatograms obtained from dosed rat plasma, blank rat plasma and EAGP solution, a total of forty-six compounds were detected in dosed rat plasma, including twenty-five prototypes and twenty-one metabolites. Among these, seventeen parent compounds were derived from Epimedium and eight were from Astragalus. These metabolites were associated with ononin (M1, M2, M9 M10 and M17), calycosin-7-<em>O</em>-<em>β</em>-D-glucoside (M6, M7, M8 and M13), icariin (M3, M4, M5, M11, M14, M15, M18, M19, M20 and M21) and methylnissolin (M12). The metabolic pathways included hydroxylation, demethylation, deglycosylation and glucuronidation. This study elucidated the potential pharmacologically active components of EAGP and provided essential data for the further study on its pharmacological materials basis and mechanism of action.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116768"},"PeriodicalIF":3.1,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143510528","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of metabolites of ganoderic acids extract from Ganoderma lucidum in rats using UHPLC–MS/MS","authors":"Wenjing Liu ,&nbsp;Nian Wu ,&nbsp;Bo Peng ,&nbsp;Ting Li ,&nbsp;Luyao Ren ,&nbsp;Bo Li ,&nbsp;Yuelin Song","doi":"10.1016/j.jpba.2025.116764","DOIUrl":"10.1016/j.jpba.2025.116764","url":null,"abstract":"<div><div>Ganoderic acids, the primary active compound cluster in <em>Ganoderma lucidum</em> (Chinese name: <em>Lingzhi</em>), one of the most reputed herbal medicines, play a crucial role in its immunomodulatory, anti-inflammatory, and anticancer properties. To reveal the existence forms of this chemical cluster in rat, we characterized the herb-derived compounds following the oral administration of the ganoderic acids extract (GAE). Moreover, we conducted metabolite characterization for both ganoderic acid A (GAA) and ganoderic acid B (GAB) in parallel to explore the metabolic pathways for ganoderic acids. The biological samples, including bile, plasma, urine, and feces, were analyzed by UHPLC–MS/MS. After carefully summarizing the mass fragmentation rules of ganoderic acids, a total of thirteen and eleven were identified after oral administration of GAA and GAB, respectively, through converting MS/MS spectra into chemical structures. Oxidation, reduction, hydroxylation, glucuronidation and sulfation were proposed as the primary biotransformation routes, and thereof, hydroxylation accounted for more metabolite generation. Through applying mass fragmentation rules and the metabolic knowledge, 107 metabolites, in total, were identified as GAE-derived components in rat. The obtained results provided important information towards the therapeutical forms of GAE <em>in vivo</em>, and moreover, offered guidelines for metabolite characterization of, but not limited to, triterpenoids.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"258 ","pages":"Article 116764"},"PeriodicalIF":3.1,"publicationDate":"2025-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143479835","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An integrated analysis strategy for characterization of chromones and coumarins from Saposhnikovia divaricata (Turcz.) Schischk. by UHPLC-QTOF-MS","authors":"Zhi-Jiang Chen ,&nbsp;Wei Guan ,&nbsp;Yan-Ying Li ,&nbsp;Yu-Qing Wang ,&nbsp;Peng Jiang ,&nbsp;Yan Sun ,&nbsp;Zhi-Chao Hao ,&nbsp;Qing-Shan Chen ,&nbsp;Li-Li Zhang ,&nbsp;Shu Liu ,&nbsp;Hai-Xue Kuang ,&nbsp;Si-Tong Liu ,&nbsp;Yao-Xin Sui ,&nbsp;Bing-You Yang ,&nbsp;Yan Liu","doi":"10.1016/j.jpba.2025.116758","DOIUrl":"10.1016/j.jpba.2025.116758","url":null,"abstract":"<div><div>This study aimed to employ a comprehensive data screening strategy to identify the chromones and coumarins present in <em>Saposhnikovia divaricata</em> (SD). Initially, the five-point mass defect filter (MDF) method was utilized to screen the respective three subclasses of chromones and coumarins for MS¹. In comparison to the traditional MDF method, the number of interference peaks was reduced from 3462 to 1053, representing a decrease of 69.58 %. Then, diagnostic fragment ion filtering (DFIF) was used to screen product ions selected by MDF method, which was based on the fragmentation rules of each subclass to screen whether it met the conditions. Finally, we characterized 94 compounds from SD, including 40 chromones and 54 coumarins, among them, 82 chromones and coumarins were identified by combining the above two methods, 12 coumarins were identified by combining MDF and references, they were classified into other coumarins. Twenty one compounds were identified for the first time from SD, and 3 chromones and 7 coumarins were unknown compounds. Through untargeted metabolomics analysis of SD samples from 12 different regions, significant differences were found in SD samples from different areas, and 20 differential metabolites were distinguished, including 13 chromones and 7 coumarins. This study established for the first time a comprehensive strategy combining MDF, DFIF, and untargeted metabolomics to evaluate SD quality. The results indicated that this method is an efficient, accurate, and promising approach for classifying and exploring compounds in complex natural product systems, providing a basis for evaluating the quality of SD from different sources.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116758"},"PeriodicalIF":3.1,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143465113","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of Astragalus polysaccharide combined with cisplatin on exhaled volatile organic compounds as biomarkers for lung cancer and its anticancer mechanism","authors":"Wenmin Shi ,&nbsp;Huanqing Zhang ,&nbsp;Hanxiao Tang ,&nbsp;Weisheng Feng ,&nbsp;Zhijuan Zhang","doi":"10.1016/j.jpba.2025.116759","DOIUrl":"10.1016/j.jpba.2025.116759","url":null,"abstract":"<div><div>Cisplatin (DDP) is widely used to fight lung cancer, but there is a risk of immune damage. <em>Astragalus</em> polysaccharides (APS) is the main active component of <em>Astragalus membranaceus</em> Bunge. It has demonstrated anticancer properties across a range of cancer types as well as to be effective against cisplatin induced immune damage. However, its therapeutic mechanism has not been fully explored. This study aimed to explore the antitumor mechanisms of APS and elucidate the relationship between APS and volatile organic compounds (VOCs) in exhaled breath of Lewis lung cancer (LLC) mice. Gas chromatography-mass spectrometry (GC-MS) was utilized to analyze the exhaled VOCs in LLC mice. A specific group of VOCs was identified as potential biomarkers for monitoring tumor progression. Furthermore, the effects of combined treatment with APS and DDP on the concentration of exhaled VOCs in LLC mice was evaluated. Stoichiometric analysis revealed that the levels of 12 VOCs exhibited substantial recovery following APS treatment. And a high concentration of APS (400 mg/kg), when combined with DDP, exhibited enhanced antitumor efficacy. The metabolic pathways involved in the action of APS include 12 pathways. Our methodology elucidated both the effects and mechanisms of APS on lung cancer, as well as the pharmacological enhancement of cisplatin by APS. These findings facilitate real-time monitoring of lung cancer treatments and contribute to the future development of anticancer therapies.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116759"},"PeriodicalIF":3.1,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143509679","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolomic machine learning predictor for arsenic-associated hypertension risk in male workers","authors":"Youyi Wu ,&nbsp;Guoliang Li ,&nbsp;Ming Dong ,&nbsp;Yaotang Deng ,&nbsp;Zhiqiang Zhao ,&nbsp;Jiazhen Zhou ,&nbsp;Simin Xian ,&nbsp;Le Yang ,&nbsp;Mushi Yi ,&nbsp;Jieyi Yang ,&nbsp;Yue Hu ,&nbsp;Xinhua Li ,&nbsp;Ping Chen ,&nbsp;Lili Liu","doi":"10.1016/j.jpba.2025.116761","DOIUrl":"10.1016/j.jpba.2025.116761","url":null,"abstract":"<div><div>Arsenic (As)-induced hypertension is a significant public health concern, highlighting the need for early risk prediction. This study aimed to develop a predictive model for occupational As exposure and hypertension using metabolomics and machine learning. A total of 365 male smelting workers from southern regions were selected. Forty workers from high and low urinary arsenic (U-As) exposure groups were chosen for non-targeted metabolomics analysis. Univariate analysis revealed that U-As is a risk factor for blood pressure and hypertension (<em>P</em> &lt; 0.05). Restricted cubic spline (RCS) analysis showed that both systolic and diastolic blood pressure, as well as hypertension risks, increased with U-As, with a threshold at 32 µg/L. Of 1145 metabolites, 383 differentially expressed metabolites (382 upregulated, 1 downregulated) were identified. Least absolute shrinkage and selection operator (LASSO) regression was used to construct a predictive model for occupational hypertension, with N-hexosyl leucine, myristic acid, gamma-glutamylvaline, and pregnanediol disulfate as predictors. The area under the curve (AUC) of the receiver operating characteristic (ROC) for the predictive model was 0.917, indicating strong predictability and accuracy. This model, based on metabolomics and machine learning, provides an effective tool for early identification and intervention for occupational populations at high risk of hypertension due to As exposure.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116761"},"PeriodicalIF":3.1,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143521020","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analytical validation and pilot clinical application of a UPLC-MS/MS method for determining intracellular mycophenolic acid and metabolites in kidney transplant recipients","authors":"Xiaomei Chen ,&nbsp;Xinhua Dai ,&nbsp;Huan Xu ,&nbsp;Chunxia Chen ,&nbsp;Xueqaio Wang ,&nbsp;Yuangao Zou ,&nbsp;Hanjing Liu ,&nbsp;Yunying Shi ,&nbsp;Yi Li ,&nbsp;Yangjuan Bai","doi":"10.1016/j.jpba.2025.116748","DOIUrl":"10.1016/j.jpba.2025.116748","url":null,"abstract":"<div><div>There is no consensus on the strategy for therapeutic drug monitoring of the immunosuppressive drug mycophenolic acid (MPA) in organ transplant recipients. The present study proposes the utilization of ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for determining the concentrations of MPA and its metabolites: 7-O-mycophenolic acid glucuronide (MPAG) and acyl mycophenolic acid glucoside (AcMPAG) in peripheral blood mononuclear cells (PBMCs). We aimed to assess the potential application of monitoring MPA and its metabolite concentrations in PBMCs in the infection after transplantation in Chinese kidney transplant recipients (KTRs). The UPLC-MS/MS method we developed demonstrated good linearity in the quantitative ranges of 0.05–50.00 ng/mL for MPA, 0.50–50.00 ng/mL for MPAG, and 0.10–20.00 ng/mL for AcMPAG. AcMPAG in PBMCs was unstable, degrading significantly after 48 h of storage at −80°C or after 3 freeze-thaw cycles. MPA and MPAG concentrations in KTRs' PBMCs exhibited high inter-individual variability, and the MPA concentration in PBMCs was poorly correlated with that in plasma (rs = 0.206, p = 0.117). Compared with the stable group, the infected group had significantly higher MPA concentration in PBMCs at 2 and 4 h post-dosing and in plasma at 4 h post-dosing (p &lt; 0.05). The receiver operating characteristic (ROC) analysis for post-transplantation infection revealed that PBMCs MPA-C4 and PBMCs-MPA-C2 possessed much better diagnostic efficiency than Plasma-MPA-C4. This method is easy-to-use and reliable, making it a promising clinical quantitative tool for MPA, MPAG, and AcMPAG in PBMCs. PBMC-MPA monitoring may be a potential biomarker for infection monitoring for KTRs.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116748"},"PeriodicalIF":3.1,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143465537","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and validation of stability-indicating UHPLC-UV-MS tandem methods for lenalidomide assay, related substances, and genotoxic impurity monitoring","authors":"Çağan Ağtaş ,&nbsp;Esen Bellur Atici","doi":"10.1016/j.jpba.2025.116757","DOIUrl":"10.1016/j.jpba.2025.116757","url":null,"abstract":"<div><div>Lenalidomide, a potent immunomodulatory drug, is widely used in the treatment of myelodysplastic syndrome. To investigate its degradation behavior under stress and stability testing conditions, we developed and validated a novel stability-indicating ultra-performance liquid chromatography (UHPLC)-UV-MS tandem method with high specificity, precision, accuracy, and robustness. An Acquity UPLC Phenyl column (100 × 2.1 mm, 1.7 µm) was used for impurity profiling and quantification of lenalidomide at a detection wavelength of 254 nm, with an injection volume of 1.0 µL and controlled sample and column temperatures of 15 °C and 30 °C, respectively. The diluent consisted of 0.1 % formic acid and acetonitrile (90:10, v/v), while the mobile phases were 0.1 % formic acid (Mobile Phase A) and acetonitrile (Mobile Phase B). A 20-minute gradient elution at a flow rate of 0.2 mL/min was used for impurity analysis, whereas a 7-minute gradient at 0.3 mL/min was applied for assay determination. The method demonstrated good linearity for all analytes, ensuring reliable quantification. Stress and photostability studies revealed that lenalidomide was stable under high temperatures (105 °C for 10 days) and daylight/UV exposure but exhibited significant degradation under hydrolytic and oxidative conditions. Hydrolysis led to the formation of major degradation products A, B, and E, whereas oxidative stress conditions generated impurities C (–NH₂ → –NO₂) and I (–NH₂ → –NH–OH). Methanol, commonly used in lenalidomide synthesis and analytical methods, was found to play a critical role in impurity formation. Methanolysis products J and K were identified as constitutional isomers arising from the ring-opening of the glutarimide moiety, which was confirmed by UHPLC-Q-TOF-MS and NMR analyses. The UHPLC-UV-MS method also reliably monitored the potentially genotoxic impurity G, classified as a Class 2 impurity according to ICH M7 guidelines, ensuring its levels remained below the Toxicological Threshold Concern (TTC, 1.5 µg/day, 60 ppm) for patient safety as per health authority requirements. This comprehensive analytical approach not only ensures the stability and safety of lenalidomide but also provides critical insights into its degradation pathways. The findings contribute to improved impurity control strategies, manufacturing process optimization, and regulatory compliance, benefiting the broader class of glutarimide-containing drug substances such as pomalidomide and thalidomide.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"258 ","pages":"Article 116757"},"PeriodicalIF":3.1,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143445630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A label-free electrochemical biosensor for rapid quantification of antimicrobial peptides in teleost fish mucus","authors":"Sonia Fekih-Zaghbib ,&nbsp;Sayda Dhaouadi ,&nbsp;Hiba Mejri ,&nbsp;Marwa Meftah ,&nbsp;Houyem Abderrazek ,&nbsp;Khaled Miled ,&nbsp;Zakaria Benlasfar ,&nbsp;Nadia Cherif ,&nbsp;Saloua Sadok ,&nbsp;Andrea Santulli ,&nbsp;Noureddine Raouafi ,&nbsp;Balkiss Bouhaouala-Zahar","doi":"10.1016/j.jpba.2025.116749","DOIUrl":"10.1016/j.jpba.2025.116749","url":null,"abstract":"<div><div>We used a thiol-faradaic electrochemical differential pulse voltammetry and impedance spectroscopy on a gold-modified screen-printed carbon electrode to quantify Chrysophsins antimicrobial peptides in the fish mucus without prior extraction. We have developed a specific anti-Chrysophsins polyclonal antibody and used ferrocene as a transducing system. The platform has a sensitivity of 30.5 nA.mL.ng⁻¹ (11.28 nA.nM⁻¹) in a linear range of 0.1–1 µg.mL⁻¹ and a limit of detection of 0.227 ng.mL⁻¹ (84.15 pM). Selectivity, accuracy, repeatability and stability were validated to meet the guidelines of ligand binding assays. Mean Chrysophsins levels in mucus pools from healthy and thermally unstressed <em>Argyrosomus regius</em>, <em>Dicentrarchus labrax</em> and <em>Sparus aurata</em> fish were 8.763 µM (± 0.007), 7.296 µM (± 0.023) and 8.296 (± 0.044) respectively, within the range of mass spectrometry gill values and below the minimum inhibitory concentration (MIC) of Chrysophsins for fish pathogens. The multi-infected <em>D. labrax</em> pool shows a significant decrease in concentration compared to the healthy and thermally stressed pools (p &lt; 0.0262) with 2.8 µM (± 0.024). The thermally stressed <em>A. regius</em> pool is not significantly different from the other pools with 8.763 µM (± 0.007). This electrochemical platform is a flexible tool for real-time targeting of peptide biomarkers in the real matrix and is suitable for mucosal fluids for early fish welfare monitoring.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"258 ","pages":"Article 116749"},"PeriodicalIF":3.1,"publicationDate":"2025-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143453905","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}