Journal of Chromatography B最新文献

{"title":"Targeted analysis of seven selected tryptophan-melatonin metabolites: Simultaneous quantification of plasma analytes using fast and sensitive UHPLC–MS/MS","authors":"Michal Kaleta ,&nbsp;Bhagya S. Kolitha ,&nbsp;Ondřej Novák ,&nbsp;Farshid Mashayekhy Rad ,&nbsp;Jonas Bergquist ,&nbsp;S.J. Kumari A. Ubhayasekera","doi":"10.1016/j.jchromb.2025.124520","DOIUrl":"10.1016/j.jchromb.2025.124520","url":null,"abstract":"<div><div>Tryptophan-derived metabolites, a group of neurotransmitters essential for various brain functions, play key roles in regulating mood, movement, sleep, and cognition. However, the comprehensive characterisation of tryptophan-melatonin pathway metabolites is challenging due to factors such as their structural diversity, chemical complexity, low concentrations, and instability of these metabolites. In this study, we developed and validated an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC–MS/MS) methodology with electrospray ionisation for the simultaneous separation and quantification of tryptophan metabolites in human plasma. The analytical calibration ranges in plasma were 0.50–200 ng/mL for serotonin, 0.01–5 ng/mL for <em>N</em>-acetylserotonin, 0.01–20 ng/mL for tryptamine, 0.01–20 ng/mL for 6-sulfatoxymelatonin, 0.01–20 ng/mL for 6-hydroxymelatonin, 0.01–100 ng/mL for melatonin, and 0.10–20 ng/mL for <em>N</em>-acetyltryptamine, with correlation coefficients ranging from 0.954 for <em>N</em>-acetyltryptamine to 0.997 for tryptamine. The intraday and interday precision remained consistently below 15 % for all analytes. Most analytes met the accuracy criteria, except for <em>N</em>-acetyltryptamine at the lowest quality control level (0.2 ng/mL), where the intraday and interday accuracy were 22.4 % and 17.4 %, respectively. In conclusion, this novel method allows for rapid identification of tryptophan-melatonin pathway intermediates in less than ten minutes, including seven distinct melatonin-related analytes. This suggests that it may find use in everyday clinical and scientific endeavours.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1256 ","pages":"Article 124520"},"PeriodicalIF":2.8,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143687820","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":"Integrated cell metabolomics and network pharmacology approach deciphers the mechanisms of Astragali Radix MIX in repairing podocyte injury","authors":"Aiping Li ,&nbsp;Xiaoyu Zhang ,&nbsp;Zheng Ju ,&nbsp;Tingting Luo ,&nbsp;Ting Cui ,&nbsp;Xuemei Qin ,&nbsp;Guangzhen Liu","doi":"10.1016/j.jchromb.2025.124522","DOIUrl":"10.1016/j.jchromb.2025.124522","url":null,"abstract":"<div><div>To elucidate the molecular mechanisms of the MIX (combined in proportion to the content in Astragali Radix (AR), named the MIX) repairing podocyte damage to ameliorate nephropathy. MTT assay and western blot analysis were used to evaluate the protective effects of MIX on MPC5 cells induced by Adriamycin (ADR). Screening of potential pharmacodynamic markers, relevant drugs and disease targets were conducted by using metabolomics combined with bioinformatics, and the most relevant metabolic pathways were identified by analyzing shared target and KEGG pathways. The key mechanism was subsequently validated by cell adhesion assays, western blot assays, and immunofluorescence staining.</div><div>The results showed that the MIX has the capacity to repair adriamycin-induced damage in MPC5 cells, as evidenced by enhanced cell viability and synaptopodin expression. Additionally, the MIX shows promise in potentially reinstating the podocyte adhesion through the modulation of the expression of podocyte adhesion-related proteins linked to nucleotide metabolites. The MIX has the potential to beneficially affect podocyte injury by modulating the cell adhesion pathway, contributing to one of the pharmacodynamic mechanisms of AR treatment of nephrotic syndrome. This has implications for the development and utilization of AR resources and achievement the social benefit of empowering rural revitalization.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1255 ","pages":"Article 124522"},"PeriodicalIF":2.8,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143403407","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 dilute–and–shoot LC–MS/MS determination of low–dosage third–generation antipsychotics and their metabolites in urine using an ultra-short column","authors":"Jeong Eun Kim,&nbsp;Seon Yeong Kim,&nbsp;Jae Chul Cheong,&nbsp;Jin Young Kim","doi":"10.1016/j.jchromb.2025.124523","DOIUrl":"10.1016/j.jchromb.2025.124523","url":null,"abstract":"<div><div>Third–generation atypical antipsychotics, known for their enhanced efficacy and reduced side effects compared to previous generations, are now extensively utilized in the treatment of schizophrenia. Due to their chemical properties and low dosages, these drugs are present at low concentrations in urine, making it challenging to monitor medication compliance among probationers. In this study, a liquid chromatography-tandem mass spectrometric (LC–MS/MS) method was developed and validated for the determination of three third-generation antipsychotics and their main metabolites in urine. A dilute-and-shoot approach was employed for rapid urine sample preparation. All compounds were separated on an ultra-short column (2.1 × 5 mm, 1.7 μm) and detected rapidly within a span of two minutes, thereby enhancing the efficiency in handling increased workloads. The limits of detection ranged from 0.01 to 0.23 ng/mL for all compounds, with correlation coefficients exceeding 0.997. The analytical method was validated using various parameters, including selectivity, precision and accuracy, matrix effect, and stability, ensuring its reliability for forensic applications. This newly developed LC–MS/MS method was successfully applied to analyze 86 urine samples obtained from probationers undergoing antipsychotic medication. Consequently, this method proves to be useful in verifying medication compliance among probationers, and effectively managing the recent increase in the number of urine drug testing.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1255 ","pages":"Article 124523"},"PeriodicalIF":2.8,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143419672","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":"Estimation of protein aggregates in Darbepoetin alfa formulations by developing a single validated analytical method","authors":"Nupur Garg,&nbsp;Sanjay Mendiratta,&nbsp;Gurminder Bindra,&nbsp;Sakshi Gehlot,&nbsp;Krutika Goswami,&nbsp;Charu Mehra Kamal","doi":"10.1016/j.jchromb.2025.124518","DOIUrl":"10.1016/j.jchromb.2025.124518","url":null,"abstract":"<div><div>Darbepoetin alfa is a new generation recombinant human erythropoiesis stimulating agent having extended half-life as compared to recombinant human erythropoietin. It is given as a life-saving drug for anemic disorders caused by chronic kidney diseases, cancer, etc. With the presence of commercial formulations of darbepoetin biosimilars in the market and non-availability of any established method in literature or testing guidelines in any compendia for their purity and quality, this research aims to develop a sensitive and robust method to determine the aggregate impurities by using size exclusion chromatographic technique. The method was optimized for sharp and high-resolution monomer peak of darbepoetin and other interferences which was further validated according to ICH Q2(R2) guidelines. The developed method was further tested on the commercial samples to verify the same. Additionally, stress testing of samples was done to simulate the after-production exposure and stability assessment. The results demonstrated that the method for analysing aggregates in darbepoetin is complying the criteria of being sensitive, specific, accurate, robust and reliable.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1254 ","pages":"Article 124518"},"PeriodicalIF":2.8,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143394585","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":"Salting-out assisted liquid-liquid extraction for UPLC-MS/MS determination of bile acids and kynurenine-, indole- and serotonin-pathway metabolites of tryptophan in human serum of healthy probands","authors":"Celine Oanes ,&nbsp;Marina Alexeeva ,&nbsp;Kjetil Søreide ,&nbsp;Cato Brede","doi":"10.1016/j.jchromb.2025.124519","DOIUrl":"10.1016/j.jchromb.2025.124519","url":null,"abstract":"<div><div>The bacterial composition of the gut has been found to affect many diseases, including several gastrointestinal cancers. The microbiome appears central in the production of certain metabolites that enter circulation, especially those from bile acids and the essential amino acid tryptophan. The tumor-microenvironment may also produce changes in metabolites, such as those from the tryptophan-kynurenine pathway, of which several compounds may be measured in the blood. As data emerges from large scale metabolomics studies, there will be a need to validate metabolomic biomarkers to confirm their clinical utility. This task also requires knowledge about biological variation of the same metabolites in a healthy population. For this purpose, a novel method was developed for quantification of bile acids and tryptophan metabolites in samples of human serum by ultra-performance liquid chromatography coupled with tandem mass spectrometry. Salting-out assisted liquid-liquid extraction was optimized with the ion-pairing reagent trifluoroacetic acid. In this way, both polar tryptophan metabolites and non-polar bile acids could be extracted with a high recovery, favorable matrix effects, and improved chromatographic focusing, by using straightforward robot pipetting. The instrumental analysis was fast (4 min and 32 s) and with sample injections done directly from the extraction microplate. The method was applied to quantify metabolites in serum from healthy probands, and for investigating inter- and intraindividual variations over six hours.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1255 ","pages":"Article 124519"},"PeriodicalIF":2.8,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143419679","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":"Uncovering the antidepressant active ingredients and related molecular mechanisms of Xiaoyao Pill using integrated pharmacological strategy","authors":"Siqian Zhou ,&nbsp;Yajing Wang ,&nbsp;Mingxia Xie ,&nbsp;Jing Li ,&nbsp;Pan Meng ,&nbsp;Juan Wu ,&nbsp;Lin Jiang ,&nbsp;Hongping Long","doi":"10.1016/j.jchromb.2025.124502","DOIUrl":"10.1016/j.jchromb.2025.124502","url":null,"abstract":"<div><h3>Purpose</h3><div>To investigate antidepressant active ingredients of XYP (Xiaoyao Pill), while predicting its primary pharmacodynamic material basis and underlying mechanisms of action.</div></div><div><h3>Methods</h3><div>UPLC-Q-TOF-MS/MS was used to identify the active ingredients of XYP. In addition, based on the analysis of components, network pharmacology and molecular docking were used to investigate potential therapeutic targets and possible signaling pathways of XYP in the treatment of depression.</div></div><div><h3>Results</h3><div>A total of 102 chemical components, 10 prototype components and 16 metabolites absorbed in the brain were identified in XYP. Network pharmacology analysis showed that these compounds shared 420 common targets with depression, TP53, EGFR, PTGS2, ESR1, PPARG and other 68 targets were considered as core targets, mainly enriched in PI3K-Akt and MAPK signaling pathways. GO analysis unveiled associated apoptosis and inflammatory response. Molecular docking revealed that paeoniflorin, liquiritin, and atractylenolide III were found to have the highest binding energy to TP53, ESR1 and PPARG.</div></div><div><h3>Conclusion</h3><div>These findings suggested that XYP may exert antidepressant effects through atractylenolide III, paeoniflorin, saikosaponin D, liquiritin, formononetin, affecting the PIK3/AKT signaling pathway. This lays the foundation for the research on the quality standards and clinical rational application of traditional Chinese medicine formulas.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1255 ","pages":"Article 124502"},"PeriodicalIF":2.8,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143479326","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":"An exploratory SWATH plasma proteomics analysis of phyllodes tumor- a type of female breast tumor","authors":"M.D. Quasid Akhter ,&nbsp;Munish Dwivedi ,&nbsp;Shivani Chitkara ,&nbsp;Navneet Kaur ,&nbsp;Swarnendu Bag","doi":"10.1016/j.jchromb.2025.124508","DOIUrl":"10.1016/j.jchromb.2025.124508","url":null,"abstract":"<div><h3>Objective</h3><div>Phyllodes tumors (PT) are rare fibroepithelial breast tumors with poorly understood molecular pathology. This study intent to identify the potential plasma markers of phyllodes tumors compared to controls using Sequential window acquisition of all theoretical fragment ion spectra-mass spectrometry (SWATH-LC-MS) based proteomics approach.</div></div><div><h3>Method</h3><div>Plasma samples from phyllodes tumor cases and controls underwent SWATH-LC-MS/MS based untargeted proteomics analysis. Proteins with 1.5 fold changes &amp; <em>p</em> &lt; 0.05 in PT cases compared to control were considered for further analysis. Statistical analysis was done by using R 4.3.1 software and proteomics analysis was performed by using Spectronaut Software</div></div><div><h3>Result &amp; conclusion</h3><div>Three hundred and nineteen proteins were identified. Amongst them 30 proteins were significantly altered in PT case compared to control. 26 were upregulated and 4 were downregulated. Again Protein-Protein network analysis revealed that 21 proteins were matched with STRING data base and out of 21 proteins 19 were highly connected in the interaction analysis. As per our knowledge, this is the first exploratory study on LC-MS/MS-SWATH based phyllodes tumor proteomics. Different proteins like APMAP, HGFAC,TTR, PNO3 etc., and associated pathways like FCGR3A-mediated IL10 synthesis, arylesterase activity, EMT related Wnt/β-catenin Pathway etc. were significantly altered in Phyllodes tumor cases. Hence. this study will help to find the plausible theranostic markers and druggable targets for the phyllodes tumors in future.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1254 ","pages":"Article 124508"},"PeriodicalIF":2.8,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143377759","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":"Residual and pharmacokinetic behavior of berberine in Carassius auratus under intraperitoneal injection conditions by HPLC-Q-TOF/MS","authors":"Shuai Zhang ,&nbsp;Le Yang ,&nbsp;Hao Jin ,&nbsp;Yuxiang Wang ,&nbsp;Qiaoqiao Teng ,&nbsp;Qi Meng ,&nbsp;Zhiqiang Cai","doi":"10.1016/j.jchromb.2025.124507","DOIUrl":"10.1016/j.jchromb.2025.124507","url":null,"abstract":"<div><div>The present study investigated the distribution characteristics of Berberine (BBR) in <em>Carassius auratus</em> under artificial environmental conditions (temperature: 26 ± 1.0 °C; continuous air pumping) following a single administration via intraperitoneal injection of an appropriate dosage of 2.0 mg/Kg per body weight. Additionally, studies have investigated the residual distribution of BBR in eight different tissues using high performance liquid chromatography-quadrupole-time of flight mass spectrometry (HPLC-Q-TOF/MS) technology. A total of 36 <em>Carassius auratus</em> (3 parallel samples per group) were in the drug-exposed group and 3 were in the control group. <em>Carassius auratus</em> were sampled at the indicated times (sampling interval: 24–96 h), lasting for approximately one month. The residual metabolic drug concentration-time curves were plotted according to the concentration of BBR in different tissues.</div><div>The maximum BBR concentrations (<em>C</em>_<em>max</em>) in all tissues were achieved 24 h after the administration of the intraperitoneal injection. The order of the <em>C</em>_<em>max</em> was: muscle &lt; eye &lt; gill &lt; brain &lt; kidney &lt; intestine&lt; liver &lt; bile. According to the distribution characteristics of BBR between tissues, the drug concentrations in muscle, brain, eye, and gill tissues were relatively low (∼100 to &gt;400 ng/g). Their metabolism was rapid, and BBR residue was significantly reduced to tens of ng/g from 24 to 200 h. Meanwhile, the bile, kidney, intestine, and liver contained significantly higher concentrations of BBR (maintained at 1200 to 6000 ng/g). The concentration shows a fluctuating and decreasing characteristic, with the drug remaining for a longer period. Following the 31 days BBR withdrawal period, the pharmacokinetic parameters of maximum observed concentration(<em>C</em>_<em>max</em>), terminal half-life (<em>T</em>_<em>1/2</em>), elimination rate constant during terminal phase (<em>λ</em>_<em>z</em>), the volume of distribution (Vd/F), and total body clearance (Cl/F) were calculated employing non-compartmental analysis (NCA) using the PK Solver software. The research results of the article explain the retention of BBR in <em>Carassius auratus</em> until excretion and can be used to determine residual levels over time.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1254 ","pages":"Article 124507"},"PeriodicalIF":2.8,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143372031","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":"Identification and characterization of the absorbed components and metabolites of Gandouling tablets in rats' plasma, liver, and urine by UPLC-Q-TOF-MSE","authors":"Tiantian Wang ,&nbsp;Qing Zuo ,&nbsp;Ting Dong ,&nbsp;Peng Wu ,&nbsp;Shijian Cao ,&nbsp;Hongfei Wu ,&nbsp;An Zhou","doi":"10.1016/j.jchromb.2025.124503","DOIUrl":"10.1016/j.jchromb.2025.124503","url":null,"abstract":"<div><div>Gandouling (GDL), a famous proprietary Chinese medicine, has been utilized in China's clinics for decades to treat Wilson's disease. However, its metabolism in vivo needs to be clarified. In this study, Ultra Performance Liquid Chromatography-quadrupole time-of-flight mass spectrometry-tandem (UPLC-Q-TOF-MS^E) was employed to analyze the metabolic pathways of these critical components in the rat after identifying the prototypes and metabolites of GDL in the plasma, liver, and urine of both normal and copper-loaded rats. As a result, 49 components were detected in the plasma of normally administered rats, including 29 prototype compounds and 20 metabolites; and 26 components were detected in the liver of normally administered rats, including 16 prototype compounds and 10 metabolites. 43 components were detected in the plasma of copper-laden administered rats, including 25 prototype compounds and 18 metabolites; and 23 components were detected in the liver of copper-laden administered rats, including 15 prototype compounds and 8 metabolites. A total of 73 GDL-related substances were detected in the urine of rats. The study results showed that the compositions in rats' plasma, liver, and urine were similar, mainly alkaloids and anthraquinones. The alkaloid components are mainly metabolized by phase I metabolism in vivo and the metabolic pathways are methylation, demethylation, etc. The anthraquinone components are mainly metabolized by phase II metabolism in vivo, and the metabolism modes are mainly glucuronidation and sulfation. The present study comprehensively analyses the metabolic properties of GDL and sets an essential basis for further investigations on the pharmacokinetics, in vivo bioactive components, and mechanism of action of GDL.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1254 ","pages":"Article 124503"},"PeriodicalIF":2.8,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143372517","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 of an analytical method for the analysis of diflubenzuron in whole blood, plasma, and serum using liquid chromatography tandem mass spectrometry","authors":"Greg Jellick ,&nbsp;Kimberly Lohmeyer ,&nbsp;Stephan Bach","doi":"10.1016/j.jchromb.2025.124506","DOIUrl":"10.1016/j.jchromb.2025.124506","url":null,"abstract":"<div><div>The integration of free-range beef livestock into the human food chain has been continuously threatened by outbreaks of babesiosis, which is propagated by ticks that vector the protozoans that cause the disease. The protozoans are spread among livestock by certain species of ticks, known collectively as cattle fever ticks (CFT). Although eliminated from the US in the 1940s, CFT continue to infiltrate the US by crossing the US/Mexico border on stray livestock and wildlife, and infest cattle herds. Orally administered acaricides are part of the strategy to control CFT. Parallel dosing studies, utilizing oral administration of the benzoylphenyl urea (BPU) compound diflubenzuron (DFB), were conducted in cattle and white tailed deer (WTD). We developed and validated a sensitive (&lt;1 ppb) liquid chromatography/ tandem mass spectrometry (LC-MS/MS) method for DFB quantification in toxicology specimens. In fit-for-purpose testing, the method demonstrated accuracies within ±9 % of QC targets, and coefficient of variation percentages (%CV) of 14.8 % for whole blood, 9.0 % for plasma, and 8.7 % for serum. This work outlines the extraction and instrumental method for DFB analysis in these matrices. LC-MS/MS had not previously been used to detect and quantify DFB in whole blood, plasma, or serum. This is also the first method to employ carbon-13 labeled DFB (¹³C₆) as an internal standard for measuring DFB in blood matrices from bovine dosing studies<em>.</em></div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1256 ","pages":"Article 124506"},"PeriodicalIF":2.8,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143512227","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}