Journal of pharmaceutical and biomedical analysis最新文献

{"title":"Neurotransmitter analysis using liquid chromatography-electrochemical detection in the 21st century: Critical review from an analytical chemistry perspective","authors":"Joanna Orzeł ,&nbsp;Monika Herian ,&nbsp;Paweł Świt","doi":"10.1016/j.jpba.2025.117120","DOIUrl":"10.1016/j.jpba.2025.117120","url":null,"abstract":"<div><div>Neurotransmitter research (detection, identification, determination) plays a special role in neuroscience, neurochemical research, and biomedical analyses. Their type or quantity allows for a better understanding of brain function, diagnosis of neurological diseases, monitoring the effects of neuroactive substances, assessing treatment effectiveness, or conducting scientific research. For these reasons, it is essential to use analytical methods suitable to detect neurotransmitters at low levels, most often in biological samples with a complex matrix (e.g., tissue homogenates, cerebrospinal fluid). This review article focuses on the use of high-performance liquid chromatography with electrochemical detection for the detection and determination of neurotransmitters as an alternative technique to mass spectrometry, capillary electrophoresis, electrochemical methods, or biosensors. The achievements made in the 21st century are presented with particular emphasis on the point of view of analytical chemistry (highly sensitive, precise, versatile applications, low‑cost/economy, multi-analyte). The article presents and discusses the description of chromatographic methods, analytical procedure validation, and the most important validation parameters, categorized into three groups: monoamines and their metabolites, amino acids, and acetylcholine with choline. Additionally, the scope and purpose of the research conducted in the area of neurotransmitter determination, the description of the tested biological samples, the method of their preparation, and the extraction procedure are presented. A novelty is the use of a generalized evaluation of chromatographic methods using tools such as RGB Additive Color Model for Analytical Method Evaluation, and Blue applicability grade index for the evaluation of method practicality. The motivation for using this technique and future perspectives are briefly discussed.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"266 ","pages":"Article 117120"},"PeriodicalIF":3.1,"publicationDate":"2025-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144892118","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":"Data mining guided affinity ultrafiltration for rapid screening of SARS-CoV-2 3CLpro inhibitors in medicinal herbs","authors":"Liqing Wang ,&nbsp;Menghan Chen ,&nbsp;Hanxue Wang ,&nbsp;Qihui Sun ,&nbsp;Sujuan Zheng ,&nbsp;Xiaoyun Liu ,&nbsp;Yong Yang ,&nbsp;Rong Rong","doi":"10.1016/j.jpba.2025.117122","DOIUrl":"10.1016/j.jpba.2025.117122","url":null,"abstract":"<div><div>Clinical studies have demonstrated that many traditional Chinese medicines (TCM) exhibit not only antiviral effects but also efficacy in alleviating clinical symptoms of Coronavirus Disease 2019 (COVID-19). However, the pharmacologically active constituents responsible for their anti-COVID-19 efficacy remain unclear. This study aimed to establish a novel strategy for identifying active ingredients from herbs clinically used for COVID-19. An integrated approach combining data mining with Affinity Ultrafiltration (AUF) was developed. Initially, using a data mining strategy, high-frequency herbs were selected from the herbs clinically used in COVID-19. Furthermore, AUF technology was used to screen for potential bioactive components from the high-frequency herbs. The anti-COVID-19 potential of active compounds was assessed through enzyme activity assays and molecular docking, followed by validation in cellular and animal models. Data mining revealed that <em>Glycyrrhiza uralensis</em> Fisch., <em>Lonicera japonica</em> Thunb. and <em>Forsythia suspensa</em> (Thunb.) Vahl were the high-frequency herbs used in COVID-19. Five potential 3-Chymotrypsin-like protease (3CL^pro) inhibitors were screened from three herbs via AUF and identified using high-resolution mass spectrometry. Further enzymatic and cellular assays demonstrated that Licochalcone C (LCC) and Forsythiaside A (FTA) inhibited Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) replication at micromolar concentrations. Notably, FTA treatment significantly suppressed the elevation of pulmonary parameters and inflammatory mediators induced by SARS-CoV-2 nucleocapsid protein in mice. In summary, this study proposes a novel strategy integrating data mining with AUF to discover active compounds from TCM. Two components, LCC and FTA, were identified as dose-dependent inhibitors of SARS-CoV-2 Omicron strain replication in vitro.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"266 ","pages":"Article 117122"},"PeriodicalIF":3.1,"publicationDate":"2025-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144864278","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":"Unraveling the loss of lysozyme concentration after tableting","authors":"Katharina Tatjana Kopp ,&nbsp;Maarten De Beer ,&nbsp;Jody Voorspoels ,&nbsp;Guy Van den Mooter","doi":"10.1016/j.jpba.2025.117116","DOIUrl":"10.1016/j.jpba.2025.117116","url":null,"abstract":"<div><div>Therapeutic proteins are fragile compounds whose functions are dependent on their higher-order structure (HOS) formed by specific intramolecular interactions. Nevertheless, protein interactions can also impede the stability if they take place with surfaces or formulation components. Besides chemical and conformational alterations, it can lead to the reduction of free non-complexed protein concentration. These are all factors which can lower the protein’s therapeutic effect and decrease therapeutic efficacy. During the minitablet development of lysozyme a loss of more than 30 % in protein recovery after tableting was observed using a reversed-phase chromatography (RPC) assay method. The aim of the current study was to unravel the cause for the loss in measured protein concentration, as prior to and post spray drying (SD) the recovery was 98.65 ± 0.76 %. It was shown that the loss was more linked to persistent protein-excipient interactions and not to the tableting process itself. It was possible to overcome these interactions with the selection of an optimized buffer system for the analytical sample preparation. For the selection of the buffer system, protein properties such as the isoelectric point (pI) and the charge had to be considered. In case of the minitablets containing lysozyme, it was a 100 mM arginine buffer pH 7.00 100 mM NaCl 0.1 % PS20 that allowed a protein recovery of almost 95.00 % after tableting. These results highlight the importance of analytical sample preparation and the selection of a suitable solvent for protein assay determinations.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"267 ","pages":"Article 117116"},"PeriodicalIF":3.1,"publicationDate":"2025-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144896607","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 pharmacokinetics and multidimensional drug-drug interactions assessment of bufadienolides based on UPLC-QqQ-MS/MS","authors":"Jun Ren ,&nbsp;Jin Zhuo ,&nbsp;Xuejing Sun ,&nbsp;Huijie Zhang ,&nbsp;Haisheng Zeng ,&nbsp;Lei Wang ,&nbsp;Wei Wang ,&nbsp;Yijun Chen","doi":"10.1016/j.jpba.2025.117118","DOIUrl":"10.1016/j.jpba.2025.117118","url":null,"abstract":"<div><div>Bufadienolides, the primary active constituents of the traditional Chinese medicine Venenum bufonis, exhibit significant pharmacological activity. However, the potential pharmacokinetic effects of drug-drug interactions among these compounds under combination treatment have not yet been investigated. Thus, this study focused on three representative bufadienolides (resibufogenin, bufalin, and arenobufagin) as the research subjects and assessed the pharmacokinetic interactions among them under co-administration conditions. Firstly, we developed a validated UPLC-QqQ-MS/MS analytical method using digoxigenin as the internal standard (IS) and performed accurate quantification of these compounds in all samples under multiple reaction monitoring (MRM) mode. This analytical method delivered high levels of specificity, recovery, accuracy, precision, and preserved sample stability. Subsequently, pharmacokinetic data for the three compounds under both single-agent and combination treatment conditions were analyzed using non-compartmental modeling. The results demonstrated significant pharmacokinetic variability among different bufadienolide compounds, reflecting distinct structure-pharmacokinetic relationships. Under combination treatment, the pharmacokinetic parameters were significantly altered due to mutual interactions among the bufadienolide compounds. In conclusion, this study has established a drug-drug interaction network for bufadienolides, providing a scientific foundation for the rational clinical application of Venenum Bufonis-based formulations. These findings may facilitate the optimized development of multi-component therapies with cardiotonic pharmacological activity, particularly in the treatment of cardiovascular diseases.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"266 ","pages":"Article 117118"},"PeriodicalIF":3.1,"publicationDate":"2025-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144886808","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":"Oxygen deficient MoO3 nanoparticles as peroxidase substitutes, their substrate-nanozyme interactions and real time validation in human serum sample","authors":"Nikhil Y. Gangadhara ,&nbsp;Manju. B ,&nbsp;P.Kiran Kumar ,&nbsp;Avinash Krishnegowda ,&nbsp;Honnur Krishna ,&nbsp;K.S. Mahesh Lohith ,&nbsp;Ravishankar H. Sadashivanna ,&nbsp;Raghavendra Ravikumar","doi":"10.1016/j.jpba.2025.117119","DOIUrl":"10.1016/j.jpba.2025.117119","url":null,"abstract":"<div><div>Oxygen deficient molybdenum oxide nanosheet (Od-MO Nshs) was synthesised by hydrothermal method and its peroxidase-like activity was established with 3,3′,5,5′-Tetramethylbenzidine-(TMB) and <em>o</em>-phenylenediamine dihydrochloride-(OPD) as a co-substrate in presence of Hydrogen peroxide (H₂O₂) and catalytic parameters were compared with horseradish peroxidase-(HRP) enzyme. Optimization was carried out for physical and chemical parameters. Synthesised nanoparticles-(NPs) were characterized for size, shape, composition oxidation state etc. NPs crystalline size was calculated using Scherrer’s equation and Williamson-Hall plot and was found to be 18.46 nm and 36.1 nm respectively. Based on the peroxidase-like activity of Od-MO Nshs nanozymes-(NZs), a simple colorimetric method was established for detecting H₂O₂ in human serum samples. Compared to HRP, Od-MO NShs showed linear detection in the fixed time method at 48 times lower H₂O₂ concentration with TMB and 16 times and 4 times lower H₂O₂ concentration accordingly. Od-MO NShs showed 2.7- and 5.3-time lesser Michaelis–Menten constant (K_m) for H₂O₂ compared to HRP with TMB and OPD, respectively indicating higher substrate affinity. The detection and quantification limits for TMB-Od-MO NShs were 0.5 and 1.5 µM, significantly lower than TMB-HRP (9.4 µM, 28.5 mM). For OPD-Od-MO NShs, the limits were 9.4 µM, and 28.5 m, compared to 1.1 µM and 3.4 µM for OPD-Od-MO NShs. in addition, these NPs exhibited photocatalytic performance, achieving effective degradation (&gt;55 %) of selected organic dyes, highlighting their potential environmental remediation application.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"266 ","pages":"Article 117119"},"PeriodicalIF":3.1,"publicationDate":"2025-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144886810","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":"Integrative analysis of gut microbiome and serum metabolomics explores the therapeutic mechanism of tongfeng qingxiao prescription in treating gouty arthritis","authors":"Xin Huang ,&nbsp;Liekui Hu ,&nbsp;Weikang Sun ,&nbsp;Zichen Shao ,&nbsp;Weiwei Ma ,&nbsp;Qipeng Yuan ,&nbsp;Jing Liu ,&nbsp;Dongyu Wu ,&nbsp;Ling Cheng ,&nbsp;Huanan Li","doi":"10.1016/j.jpba.2025.117121","DOIUrl":"10.1016/j.jpba.2025.117121","url":null,"abstract":"<div><div>Gouty arthritis (GA), a metabolic inflammatory disorder, shows increasing global prevalence, especially in China. Conventional GA treatments often cause adverse reactions, necessitating alternative therapies. Tongfeng Qingxiao Prescription (TFQXP) demonstrates clinical efficacy in GA, though its mechanisms remain unclear. This research explored TFQXP's effects in GA rats using gut microbiome and serum metabolomics analyses. Sprague-Dawley rats (n = 36) were randomly divided into six groups: control, model, TFQXP low-dose, medium-dose, high-dose, and positive groups. GA was induced in all groups except the control group and confirmed via gait analysis and infrared thermal imaging. Inflammation was assessed by measuring ankle joint edema. Histopathology evaluated colon and ankle joint tissues, while transmission electron microscopy examined intestinal epithelium. Mucin 2 expression was analyzed via immunohistochemistry. Synovial TLRs/MyD88/NF-κB pathway components were evaluated using western blotting and qRT-PCR. Gut microbiota structure was assessed via 16S rDNA sequencing, while serum metabolites were analyzed using non-targeted metabolomics. Multi-omics correlation analyses explored gut microbiota-serum metabolite relationships. TFQXP appears to mitigate GA-induced synovial and cartilage damage by suppressing the TLRs/MyD88/NF-κB signaling pathway, resulting in downregulation of pro-inflammatory mediators. Furthermore, TFQXP promoted intestinal barrier repair, alleviated dysbiosis, and ameliorated metabolic pathway dysregulation. Thus, TFQXP may exert therapeutic effects on GA by modulating the \"gut-joint\" axis.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"266 ","pages":"Article 117121"},"PeriodicalIF":3.1,"publicationDate":"2025-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144890849","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":"Method development for poly(diallyldimethylammonium chloride) determination in proteinaceous samples and its role in downstream technology development of monoclonal antibody production","authors":"Zoltán Sávoly ,&nbsp;Enikő Szilágyi ,&nbsp;Dóra Kozák ,&nbsp;Viktor Vásárhelyi ,&nbsp;Hajnalka Szabados","doi":"10.1016/j.jpba.2025.117113","DOIUrl":"10.1016/j.jpba.2025.117113","url":null,"abstract":"<div><div>Poly(diallyldimethylammonium chloride) (pDADMAC) is a cationic polyelectrolyte bearing one positive charge per monomer unit. This structure enables its application as flocculating agent. It can be used in the downstream processing of biological drug production. Due to its potential toxicity, its concentration in the drug substance should be reduced to a sufficiently low level, which is realized by chromatographic steps of downstream processing. To prove proper clearance, appropriate analytical method should be developed, which should be able to measure low pDADMAC concentration in matrices with high protein content. It is a challenge due to the size and composition of pDADMAC. Its molar weight (∼300 kDa) is commensurable with that of mAbs (∼150 kDa) or rather their dimers. It hampers the applicability of size-exclusion chromatography. For reversed-phase HPLC the protein content of the samples should be removed, which is especially challenging due to electrostatic interaction between protein and the pDADMAC analyte. The development and qualification of an RP-HPLC method is presented in this study. The protein removal is realized by precipitation using ammonium sulfate, which is a quick and simple method. The method was found to be able to measure pDADMAC concentration accurately and precisely in samples with up to 85 g/L protein concentration.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"268 ","pages":"Article 117113"},"PeriodicalIF":3.1,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144865489","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":"Species-specific identification of pangolin scales using UPLC/Q-TOF-MS with LC-MS/MS validation","authors":"Jinju Zhang ,&nbsp;Lidan Zhang ,&nbsp;Jungang Lu ,&nbsp;Xianlong Cheng ,&nbsp;Wenyan Zheng ,&nbsp;Feng Wei ,&nbsp;Xiaoxiao Liu","doi":"10.1016/j.jpba.2025.117115","DOIUrl":"10.1016/j.jpba.2025.117115","url":null,"abstract":"<div><div>Pangolin scales (PS) are derived from various pangolin species and are commonly subjected to thermal processing. Identifying their species of origin using traditional methods is challenging. Therefore, a reliable approach for species identification of PS and their processed products is urgently needed. In this study, a method combining ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS) with principal component analysis (PCA) was developed to distinguish PS from three different species. Seven species-specific ions were identified and demonstrated good specificity for their corresponding species. Based on these markers, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) multiple reaction monitoring (MRM) method was established, enabling rapid species identification of PS and their processed products within 20 min. The method was successfully applied to 58 PS samples of unknown origin, demonstrating high reliability and applicability. This study provides an effective approach for the authentication and quality control of PS and their processed products.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"268 ","pages":"Article 117115"},"PeriodicalIF":3.1,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144865488","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 improved pretreatment method for efficient detection of antibiotic residues in aquatic products by LC-MS/MS","authors":"Yu Qin ,&nbsp;Shenping Zhang ,&nbsp;Jingwen Shi ,&nbsp;Heng Gong","doi":"10.1016/j.jpba.2025.117114","DOIUrl":"10.1016/j.jpba.2025.117114","url":null,"abstract":"<div><div>Antibiotics are a class of compounds that are effective against bacterial infections and are widely used in aquaculture. Herein, a method based on high performance liquid chromatography coupled with tandem mass spectrometry was developed to detect nine classes of 116 antibiotics in aquatic products. The antibiotics were extracted in two steps using 1 % formic acid-acetonitrile with 0.1 mol/L Na₂EDTA-Mcllvain buffer solution and acetonitrile, purified using Captiva EMR-Lipid, and separated with an ACQUITY UPLC®HSS T₃ column (100 mm × 2.1 mm, 3.5 μm) using acetonitrile and 0.1 % formic acid with 5 mM ammonium acetate as mobile phase for gradient elution. Calibration curve from 0.10 to 100 ng/mL exhibited good correlation coefficient (R²&gt;0.99) for quantification. The analytical performance of the developed method was satisfactory, with limits of quantification ≤ 5.00 ng/g for detecting of antibiotics in aquatic products. The recoveries of the 116 antibiotics ranged from 60.57 % to 127.33 %, with precision (expressed as relative standard deviation) of &lt; 30.20 %. All parameters showed acceptable values and conformed to the Commission Decision 2021/808/EC criteria. When compared with other techniques, the developed method meets the quantitative requirements of high-throughput analysis and can simultaneously analyze 116 antibiotics in aquatic products.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"266 ","pages":"Article 117114"},"PeriodicalIF":3.1,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144864276","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":"Antibody-free quantification of serum infliximab using LC-MS/MS","authors":"Xiaoshuang He ,&nbsp;Juntao Zhao ,&nbsp;Xiaoli Ma ,&nbsp;Wenkai Chen ,&nbsp;Shu Chen ,&nbsp;Kaifeng Shi ,&nbsp;Jian Wei ,&nbsp;Zhichao Gu ,&nbsp;Jie Fang ,&nbsp;Fengmei Hu ,&nbsp;Xiaolan Bian ,&nbsp;Jing Sun ,&nbsp;Yuan Xiao ,&nbsp;Qiuya Lu","doi":"10.1016/j.jpba.2025.117117","DOIUrl":"10.1016/j.jpba.2025.117117","url":null,"abstract":"<div><h3>Background</h3><div>Infliximab (IFX) is the first biologic to be approved to treat inflammatory bowel disease (IBD). Serum IFX concentrations can be correlated with the clinical prognosis of IBD. Liquid chromatography–tandem mass spectrometry (LC-MS/MS) is an alternative to clinical immunoassays owing to its higher selectivity and sensitivity. An efficient antibody-free LC-MS/MS method was developed in this study to determine serum IFX levels.</div><div>Methods</div><div>Protein G–coated magnetic beads were used to capture IFX and the internal standard (cadonilimab [CADO]) in human serum. After enrichment, IFX and CADO were digested using trypsin, and signature peptides were selected for subsequent LC-MS/MS analysis. The results were compared with those from the corresponding immunoassay.</div><div>Results</div><div>The selected peptides had good specificity, and the total precision was within 10.5 %. The accuracy was between 92.8 % and 97.6 % in the linear range of 0.5–100 mg/L. The results from the sample assay were in agreement with those from the immunoassay.</div><div>Conclusion</div><div>Determination of IFX using LC-MS/MS is more sensitive and robust than other analytical methods. Therefore, the method reported in this study shows potential for use in therapeutical drug monitoring and developing individualized treatment modalities.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"266 ","pages":"Article 117117"},"PeriodicalIF":3.1,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144890896","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}