Journal of pharmaceutical and biomedical analysis最新文献

{"title":"Importance in optimization of sample volume in the IgG-immunoaffinity UPLC-MS/MS assay for quantification of a therapeutic antibody in monkey serum","authors":"Kenji Kita ,&nbsp;Yuji Mano","doi":"10.1016/j.jpba.2026.117334","DOIUrl":"10.1016/j.jpba.2026.117334","url":null,"abstract":"<div><div>E6011 is a novel monoclonal antibody developed for the treatment of rheumatoid arthritis. Ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC–MS/MS) has become increasingly important in the assays of therapeutic antibodies, yet achieving high sensitivity remains a major challenge. For the E6011 assay, an immunoaffinity (IgG-IA) method employing protein G in combination with UPLC-MS/MS was utilized and qualified in monkey serum. Method development revealed that increasing the matrix volume did not always lead to improved sensitivity when using a fixed volume of protein G magnetic beads, due to their limited capture capacity. Using only 1.8 µL of monkey serum, E6011 was quantitatively analyzed at concentrations as low as 1 µg/mL. Accuracy and precision were within ±20 %, meeting the acceptance criteria, however, the selectivity result at 1 µg/mL highlighted the need for evaluation of selectivity across multiple individuals. E6011 concentrations in monkey serum were determined using the IgG-IA-UPLC-MS/MS method, and the resulting pharmacokinetic parameters were comparable to those previously reported using UPLC-MS/MS with ammonium sulfate precipitation and a ligand-binding assay. These findings highlight the importance of optimizing matrix volume in assay development and demonstrate the successful application of the established method in a monkey pharmacokinetic study.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117334"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145926822","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 a rapid LC-MS/MS method for methylphenidate, atomoxetine, and their metabolites with application to pediatric pharmacokinetics","authors":"Duygu Eryavuz Onmaz ,&nbsp;Merve Kuz ,&nbsp;Fatih Hilmi Cetin ,&nbsp;Halit Necmi Ucar ,&nbsp;Serhat Turkoglu ,&nbsp;Rabia Coban ,&nbsp;Ali Unlu","doi":"10.1016/j.jpba.2025.117321","DOIUrl":"10.1016/j.jpba.2025.117321","url":null,"abstract":"<div><div>Therapeutic drug monitoring of attention-deficit/hyperactivity disorder (ADHD) pharmacotherapy is challenged by substantial interindividual variability and the lack of rapid, multiplex analytical platforms. We developed and validated a fast, integrated 5-minute LC–MS/MS assay for the simultaneous quantification of stimulant (methylphenidate, ritalinic acid) and non-stimulant (atomoxetine, 4-hydroxy-atomoxetine) ADHD medications in human serum. The method employs a single-step protein precipitation and requires only 200 µL of serum. Validation demonstrated excellent linearity, precision, accuracy, minimal matrix effects, and robust analyte stability in accordance with FDA and CLSI guidelines. Application to pediatric ADHD patients revealed formulation- and dose-dependent concentration differences for methylphenidate and atomoxetine, along with strong parent–metabolite correlations. No significant associations were observed with age, sex, or body mass index. This rapid and reliable LC–MS/MS platform supports clinically actionable therapeutic drug monitoring and provides a practical framework for personalized ADHD treatment.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117321"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145819885","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 critical review of molecularly imprinted polymer technology for aflatoxin sensing: Current status and future directions","authors":"Ahmet Cetinkaya ,&nbsp;S. Irem Kaya ,&nbsp;Burcak Demirbakan ,&nbsp;Mehmet Altay Unal ,&nbsp;Mustafa Kemal Sezginturk ,&nbsp;Sibel A. Ozkan ,&nbsp;Evrim Gunes Altuntas","doi":"10.1016/j.jpba.2025.117330","DOIUrl":"10.1016/j.jpba.2025.117330","url":null,"abstract":"<div><div>Since the first discovery and identification of aflatoxins almost 70 years ago, their occurrence, potential risks, and health hazards remain of great importance. Therefore, several chromatography-, colorimetry-, and immunoassay-based analytical applications for aflatoxin detection are reported in the literature. Molecularly imprinted polymers (MIPs) have attracted considerable interest in sensor design due to their unique potential as synthetic recognition elements, providing high selectivity and excellent stability. These features help overcome the disadvantages of conventional sensing systems. The ongoing advancement of MIP-based sensors presents new opportunities for future applications and offers a reliable and versatile approach to meet the growing demands of current analytical research. This review focuses on studies published since 2023 concerning the detection of aflatoxins using MIP-based sensors, which hold a prominent and critical position in the literature. Additionally, this review addresses several limitations of MIP-based sensors, particularly in the context of aflatoxin detection, and discusses proposed solutions to overcome these challenges. Lastly, current advancements and future expectations in the topic are also highlighted.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117330"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145881328","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":"In-depth O-glycosylation characterization and comparison of commercially available etanercept biosimilars by using Electron Activated Dissociation","authors":"Wenhong Fan ,&nbsp;Xiang Li ,&nbsp;Wentao Wang ,&nbsp;Tie Gao ,&nbsp;Ji Luo ,&nbsp;Zoe Zhang ,&nbsp;Bingjie Liu ,&nbsp;Chenggang Liang ,&nbsp;Hongxu Chen","doi":"10.1016/j.jpba.2025.117326","DOIUrl":"10.1016/j.jpba.2025.117326","url":null,"abstract":"<div><div>Glycosylation is one of the important post-translational modifications in protein drugs expressed by eukaryotic cells. It is closely related to the conformation, stability, and solubility of therapeutic proteins and holds vital biological significance. Etanercept contains 26 O-glycosylation sites with high glycosylation heterogeneity, which makes the analysis of O-glycosylation highly challenging. In this paper, using liquid chromatography-mass spectrometry (LC-MS), 11 O-glycosylation on the single chain of etanercept were first identified at the intact protein level, with Core1 and Core2 O-glycans. Second, 11 O-glycosylation sites were identified with the electron activation dissociation (EAD) fragmentation mode at the glycopeptide level, and their glycans were comprehensively characterized. The major O-glycan structures were Core1 and Core2, which were consistent with those observed at the intact protein level. Third, the etanercept innovator (Y1) and six biosimilars or follow-on products (Y2-Y7) from different manufacturers were also compared. Compared with the etanercept innovator, the results showed that etanercept biosimilars or follow-on products from different manufacturers exhibited pronounced differences in O-glycosylation modification due to variation in production processes. Based on a comprehensive assessment of consistency in deconvoluted spectra and the glycosylation ratios, etanercept biosimilar Y2 was the most similar to the innovator Y1. In-depth studies of etanercept’s O-glycosylation can effectively be applied to the etanercept innovator's comparative analysis and biosimilars. Furthermore, the LC-MS method combined with the EAD fragmentation technique can offer detailed and precise data support for the comprehensive quality control of complex glycoprotein drugs and the optimization of production processes.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117326"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145881326","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":"Integrated proteomics and metabolomics reveal NF-κB pathway modulation by Xiangpi Shengji ointment and Qingre Zaoshi compound in anal fistula wound healing","authors":"Mintao Jian ,&nbsp;Jiaqing Xiong ,&nbsp;Kui Li","doi":"10.1016/j.jpba.2025.117306","DOIUrl":"10.1016/j.jpba.2025.117306","url":null,"abstract":"<div><div>This study investigates the molecular mechanisms by which Xiangpi Shengji Ointment (XPSJO), combined with a heat-clearing and dampness-drying compound, accelerates postoperative wound healing in an anal fistula (AF) rat model, employing integrated proteomic and metabolomic approaches. An AF rat model was established and treated topically with the herbal combination. Wound healing progression was monitored postoperatively. Histopathological assessment (hematoxylin and eosin [H&amp;E] staining), enzyme-linked immunosorbent assay [ELISA] quantification of inflammatory markers (e.g., tumor necrosis factor-alpha [TNF-α], interleukin [IL]-6) and vascular endothelial growth factor [VEGF], and proteomic (liquid chromatography-tandem mass spectrometry [LC-MS/MS]) and metabolomic (gas chromatography-mass spectrometry [GC-MS]) analyses identified critical pathways and biomolecules involved. <em>In vitro</em>, lipopolysaccharide (LPS)-stimulated THP-1 macrophages and human umbilical vein endothelial cells (HUVECs) were co-treated with XPSJO and <em>Coptis chinensis</em>-<em>Phellodendron amurense</em> Rupr. Extract (CGE). Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway modulation was examined via reverse transcription quantitative polymerase chain reaction (RT-qPCR), Western blot, and functional assays (cell counting kit [CCK]-8, flow cytometry, scratch wound, tube formation). Lentiviral NF-κB overexpression validated mechanistic specificity <em>in vitro</em> and <em>in vivo</em>. Results showed that XPSJO combined with the compound accelerated AF wound closuresuppressed NF-κB signaling pathway activation (evidenced by downregulation of pathway-associated proteins in proteomics and reduced p65 phosphorylation), decreased pro-inflammatory cytokine secretion, and enhanced VEGF expression. Critically, it promoted M2 macrophage polarization (increased Arg-1/CD206, decreased iNOS/CD80) and stimulated HUVEC migration/angiogenesis. Multi-omics analysis further confirmed NF-κB, TNF, and IL-17 pathways as central therapeutic targets. The study concludes that XPSJO combined with a heat-clearing and dampness-drying compound promotes AF healing through pharmacological inhibition of the NF-κB pathway, macrophage reprogramming, and tissue remodeling.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117306"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733403","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":"The role of syringe residue analysis in identifying substances consumed in sexualized drug use practices","authors":"Carlos Pernas-Fraguela ,&nbsp;Rosario Rodil ,&nbsp;José Benito Quintana ,&nbsp;Antonio Saez ,&nbsp;Ruth Olmos ,&nbsp;Joan Mestre ,&nbsp;Manuel Isorna ,&nbsp;Rosa Montes","doi":"10.1016/j.jpba.2025.117311","DOIUrl":"10.1016/j.jpba.2025.117311","url":null,"abstract":"<div><div><em>Chemsex</em> is a practice mostly extended between men who have sex with men under drugs effects to enhance and/or prolong sexual activities. Within <em>chemsex</em>, <em>slamming</em> consists of intravenous consumption of substances such as cathinones and amphetamines. Syringe residue analysis is a relatively new alternative for knowing with accuracy the substances consumed by the users. Although several studies have been carried out applying the technique, none of them was focused on this population group. In this work, syringes were obtained from a drug addiction assistance centre from Madrid (Spain). Samples were processed by rinsing with methanol and injected in a liquid chromatography coupled to high resolution mass spectrometry system. Data (acquired in data dependent mode) were processed using a suspect screening approach. In total, 122 syringes from three sampling campaigns were analyzed. The compounds belonging to the cathinones’ group were the most frequently detected (99 %), followed by amphetamines and cocaine. The most consumed cathinone was chloromethcathinone, but also others as methylmethcathinone, pentedrone or N,N-dimethylpentylone were identified. Furthermore, temporal trends were detected during the different sampling campaigns. Thus, this technique allowed us to determine in an accurate way the substances consumed during <em>slamming</em> sessions, which may differ from the self-declared ones in surveys. Moreover, temporal trends were observed, that align with the market changes and regulation of cathinones.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117311"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733401","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 MA-SSA-optimized XGBoost-MLP framework using LIBS for rapid classification and quantitative analysis of heavy metals in traditional chinese medicines","authors":"Abulimiti Yasen ,&nbsp;Yuxia Zhou ,&nbsp;Wenhan Gao ,&nbsp;Zhongxin Zhang ,&nbsp;Reyihan Tudi ,&nbsp;Mei Xiang ,&nbsp;Bumaliya Abulimiti","doi":"10.1016/j.jpba.2025.117296","DOIUrl":"10.1016/j.jpba.2025.117296","url":null,"abstract":"<div><div>Laser-Induced Breakdown Spectroscopy (LIBS) holds significant value for rapid elemental detection; however, strong spectral interference, matrix effects, and high-dimensional data characteristics pose considerable challenges to accurate quantitative analysis. To enhance the performance of LIBS quantitative analysis, this study proposes a novel machine learning framework that integrates XGBoost and Multilayer Perceptron (MLP), optimized by a Multi-dimensional Adaptive Sparrow Search Algorithm (MA-SSA). The framework employs XGBoost for automated feature selection, eliminating redundant spectral variables while retaining critical information, and utilizes MA-SSA to optimize the hyperparameters of the MLP in regression tasks, significantly improving model stability and prediction accuracy. Experimental results demonstrate that the proposed method achieves 100 % accuracy in multi-class classification, outperforming traditional classifiers such as Random Forest, XGBoost, and standalone MLP. In terms of quantitative detection, the MA-SSA-optimized model achieves an RMSE of 4.43 µg/g, surpassing other hybrid optimization models including XGBoost-SSA-MLP (RMSE=4.62 µg/g), XGBoost-PSO-MLP (RMSE=5.225 µg/g), and XGBoost-GA-MLP (RMSE=5.584 µg/g). XGBoost-based feature selection effectively reduces spectral dimensionality while maintaining predictive performance. The proposed MA-SSA algorithm further enhances convergence efficiency and generalization capability. This study provides a robust, efficient, and scalable solution for LIBS analysis, with broad application potential in the field of real-time quantitative detection.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117296"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145691109","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 In Vitro metabolic profiles of crepidatin in human liver microsomes and hepatocytes using HPLC-MS/MS and HPLC-Q-Orbitrap-HRMS","authors":"Yaqin Sun ,&nbsp;Junyin Zhou ,&nbsp;Xiaoli Hou ,&nbsp;Tianqi Ma ,&nbsp;Ye Qiao ,&nbsp;Luxiang Sun ,&nbsp;Hong Cheng","doi":"10.1016/j.jpba.2025.117269","DOIUrl":"10.1016/j.jpba.2025.117269","url":null,"abstract":"<div><div>Crepidatin, a bioactive bibenzyl derivative isolated from multiple plants of Dendrobium, has recently gathered significant interest due to its activity against lung cancer. The aim of this study was to elucidate the metabolic fates of crepidatin using human liver microsomes and hepatocytes. To achieve this goal, a specific HPLC-MS/MS method for the determination of crepidatin in human liver microsomes and hepatocytes was developed and validated. Separation was achieved within 2 min on an ACQUITY BEH C₁₈ column (1.7 μm, 2.1 × 50 mm) with a mobile phase consisting of 0.1 % formic acid in water and methanol/acetonitrile (30:70, <em>v/v</em>), running at 0.4 mL/min. Detection employed positive electrospray ionization in multiple reaction monitoring (MRM) mode, with the transition <em>m/z</em> 319.1→151.1 for both crepidatin and internal standard (erianin). The developed method displayed excellent linearity over the concentration range of 1.0–1000 nM (<em>r</em> &gt; 0.999). Crepidatin showed rapid metabolism both in human liver microsomes (<em>t</em>_1/2: 17.2 min) and hepatocytes (<em>t</em>_1/2: 10.8 min). Metabolite identification was achieved using HPLC-Q-Orbitrap-HRMS operated in full MS/dd-MS² scan mode. A total of 15 metabolites across five metabolic pathways were structurally characterized through accurate mass measurement and fragmentation analysis. Hydroxylation (M5, M7) and glucuronidation (M14) emerged as the predominant metabolic pathways. Furthermore, incubation with glutathione (GSH)-supplemented human liver microsomes led to the detection of three GSH conjugates, providing direct evidence for crepidatin bioactivation via the formation of ortho-quinone and quinone-methide species. Crepidatin was primarily metabolized by CYP3A4, 2C8, 2C19, UGT1A1, UGT1A8, and UGT1A9. This study describes the first integrated HPLC-MS/MS and HPLC-Q-Orbitrap-HRMS method for its in vitro metabolic profiling, and the results facilitate prediction of <em>in vivo</em> pharmacokinetics.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117269"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145555412","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":"Simultaneous identification of 50 illegal adulterants in dietary supplements using high-performance liquid chromatography–single quadrupole mass spectrometry","authors":"Takayuki Sakai ,&nbsp;Misa Tanaka ,&nbsp;Yuki Azuma,&nbsp;Yusuke Sakamoto,&nbsp;Takaomi Tagami,&nbsp;Akiko Asada,&nbsp;Takahiro Doi","doi":"10.1016/j.jpba.2025.117300","DOIUrl":"10.1016/j.jpba.2025.117300","url":null,"abstract":"<div><div>Health hazards associated with the ingestion of dietary supplements containing illegal adulterants have been reported. High-performance liquid chromatography<img>single quadrupole mass spectrometry (LC-MS) is useful for identification of illegal adulterants in dietary supplements because of its moderate sensitivity and wide accessibility. Previously, we reported a method for the simultaneous identification of 18 illegal adulterants in dietary supplements using LC-MS. Considering numerous analogs of pharmaceutical ingredients recently identified, developing a more convenient screening method to identify a broader range of such compounds is essential. In this study, we introduced another LC-MS-based method to simultaneously identify 50 illegal adulterants in dietary supplements, including newly identified compounds such as descarbonsildenafil and <em>N</em>-phenylpropoxyphenylcarbodenafil. The proposed method achieved clear separation of the targeted peaks in their extracted ion chromatograms. All coefficients of determination (r²) exceeded 0.9794, with most compounds showing good linearity above 0.99. Recovery tests confirmed the specificity of detection for each compound, with recovery rates ranging from 74.9 % to 127.7 % and relative standard deviations below 21.3 %. Validation results demonstrated the method’s suitability for identifying all 50 targeted compounds and quantifying 32 compounds with acceptable accuracy and precision. The method also successfully detected illegal adulterants in positive samples. Thus, the proposed method may be useful for identifying—and partially quantifying—illegal adulterants present in dietary supplements.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117300"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145714540","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":"Validation of the Bactec system for sterility testing of advanced therapy medicinal product suspensions in Ringer’s Lactate","authors":"Ilona Szabłowska-Gadomska,&nbsp;Marta Bochyńska-Czyż,&nbsp;Agnieszka Mroczko","doi":"10.1016/j.jpba.2025.117283","DOIUrl":"10.1016/j.jpba.2025.117283","url":null,"abstract":"<div><div>Ensuring sterility is a key quality requirement in the manufacturing of cell-based medicinal products. This study evaluated the capability of the BACTEC automated microbial detection system to identify viable microorganisms in adipose-derived stem cell-based products suspended in Ringer’s Lactate. The validation process comprised four stages: i) sterility testing of uninoculated media to assess reagent quality; ii) growth promotion test to confirm medium performance; iii) suitability testing for compatibility with the sample matrix; iv) confirmation using pharmacopeial methods. All inocula met the European Pharmacopoeia recommendations. Validation was defined as the demonstration that the BACTEC system provides results equivalent to the pharmacopeial sterility test in terms of sensitivity, reliability, and detection time. Evaluated parameters included accuracy, repeatability, and detection limit, following European Pharmacopoeia requirements for alternative microbiological methods. The system successfully detected microbial contamination for all reference strains with a detection limit of 5 CFU and repeatability &gt; 95 %. All microorganisms showed growth within 24–72 h, well inside the 7-day acceptance criterion. Quantitative comparison with the pharmacopeial method demonstrated full concordance (Cohen’s κ = 1.0) and a strong correlation of detection times (r = 0.92, p &lt; 0.001). These results confirm that the BACTEC system is a validated and reliable alternative to the pharmacopeial sterility test, providing faster detection without loss of analytical accuracy. The validation supports its applicability for sterility assurance of cell-based materials and integration into quality management during the manufacture of advanced therapy medicinal products.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117283"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622391","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}