Journal of Chromatography B最新文献

{"title":"Improved analysis HPLC-ESI/triple method for mapping the methotrexate by mass spectrometry","authors":"Haihong Jia ,&nbsp;Ruihong Li ,&nbsp;Yahui Li ,&nbsp;Fen Lu ,&nbsp;Lan Ma ,&nbsp;Xiujuan Xu","doi":"10.1016/j.jchromb.2025.124529","DOIUrl":"10.1016/j.jchromb.2025.124529","url":null,"abstract":"<div><div>Methotrexate is a commonly utilized agent in pediatric oncology therapy. However, significant interindividual variability in its clearance can lead to delayed clearance and resultant severe toxicity. This underscores the urgent need for efficient and sensitive analytical methods to ensure patient safety. In this study, we aimed to establish a high-performance liquid chromatography-tandem mass spectrometry (HPLC-ESI/triple) method for the quantitative determination of methotrexate concentrations in plasma. This method is intended to facilitate therapeutic drug monitoring in pediatric patients, thereby allowing for a better understanding of the pharmacokinetics of methotrexate in vivo. The results indicate that the established HPLC-ESI/triple method can accurately and sensitively quantify methotrexate using only 10 μL of plasma. The recovery rates for all analytes exceeded 90 %, and matrix effects were minimal. Furthermore, our optimized method revealed that patient age significantly influences methotrexate blood concentration. Specifically, under identical dosage and administration intervals, an increase in patient age correlates with a decrease in measured blood concentration. Additionally, our findings suggest that measuring methotrexate concentrations within 24 h post-administration enhances the effectiveness of monitoring, thereby promoting rational drug use and ensuring optimal therapeutic dosing.</div><div>In summary, we have conducted a comprehensive study establishing a robust method for determining methotrexate concentrations in patient plasma. The optimized HPLC-ESI/triple method is poised for widespread application in clinical practice to monitor methotrexate therapy in pediatric patients.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1255 ","pages":"Article 124529"},"PeriodicalIF":2.8,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143471330","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":"Evaluation of binding interaction between compounds targeting peroxisome proliferator-activated receptor γ in Nelumbinis folium using receptor chromatography and molecular dynamic simulation","authors":"Qingqing Yao ,&nbsp;Jiatai Yin ,&nbsp;Xiuli Ji ,&nbsp;Xue Li ,&nbsp;Yifan Gao ,&nbsp;Dan Lu ,&nbsp;Ying Chen ,&nbsp;Qian Li ,&nbsp;Dalong Zhi","doi":"10.1016/j.jchromb.2025.124528","DOIUrl":"10.1016/j.jchromb.2025.124528","url":null,"abstract":"<div><div>Despite considerable efforts invested in clinical trials aimed at treating obesity and enhancing the metabolic profiles of <em>Nelumbinis Folium</em>, the precise phytochemicals involved and their mechanisms of action remain unclear due to the absence of an efficient screening technique. Herein, <em>Nelumbinis Folium</em> serves as the focal point to elucidate the bioactive compounds that specifically bind to peroxisome proliferator-activated receptor γ using immobilized receptor chromatography. Following identification through liquid chromatography-mass spectrometry, the compounds were further evaluated using chromatographic techniques and molecular dynamics simulations. The results unveiled catechin and hypericin as the receptor-binding compounds present in <em>Nelumbinis Folium</em>, with hypericin exhibiting a stronger affinity and a faster dissociation rate constant compared to catechin. Molecular dynamics studies highlighted the crucial role of cysteine located at position of 285 in the receptor ligand binding domain during the initial ligand capture phase. Subsequently, Van Der Waals forces and electrostatic interactions facilitated the binding process. The calculated standard binding free energies were − 61.75 ± 2.61 kcal/mol for hypericin and − 43.19 ± 0.63 kcal/mol for catechin. Collectively, these findings provide valuable insights into receptor-drug interactions and confirm the effectiveness of immobilized receptor chromatography in screening potential lead compounds from complex systems.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1255 ","pages":"Article 124528"},"PeriodicalIF":2.8,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143463660","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":"Tissue distribution, excretion characteristics and metabolic profiling studies of oxypeucedanin in rats using liquid chromatography tandem mass spectrometry","authors":"Huixiao Duo ,&nbsp;Liyun Wang ,&nbsp;Pei Lu ,&nbsp;Xiaodan Zhang ,&nbsp;Mingcong Zheng ,&nbsp;Hanying Song ,&nbsp;Juan Zhou ,&nbsp;Wenzhuo Lei ,&nbsp;Shushu Ding ,&nbsp;Jie Jia Li ,&nbsp;Junxu Li ,&nbsp;Qing Zhu","doi":"10.1016/j.jchromb.2025.124525","DOIUrl":"10.1016/j.jchromb.2025.124525","url":null,"abstract":"<div><div>Oxypeucedanin (OPD) is a linear furanocoumarin extracted from Chinese herbs, which has been reported to exhibit various pharmacological effects such as analgesia, antitussive, anticancer and anti-inflammatory activities. In this paper, the tissue distribution, excretion and metabolism characteristics of OPD in rat was conducted in order to better understand this compound and for potential development of OPD as a medicine. We first used the established HPLC/MS/MS method to examine the plasma protein binding rate and the tissue distribution of OPD in rats after oral administration. The excretion characteristics of OPD in urine, feces and bile, as well as its metabolic profiles in plasma, urine and bile were also investigated. The results demonstrated that after OPD administration, extensive metabolism occurred in rats, involving first-pass effect and enterohepatic circulation, with significant sex differences observed in the metabolic process. The study identified eight metabolites in rats, indicating that OPD metabolism primarily occurs through oxidation, reduction, and glucuronidation. In summary, this study systematically examined the basic process of OPD in vivo, providing a robust foundation for accurately predicting the behavior, efficacy and safety of OPD in humans.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1255 ","pages":"Article 124525"},"PeriodicalIF":2.8,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143474198","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 comprehensive evaluation of a bioanalytical technique for Encorafenib and Cetuximab combination Cancer therapy by LC-MS/MS and their pharmacokinetics in plasma","authors":"Anoop Bodapati ,&nbsp;Bangaraiah Pagala ,&nbsp;Sudha Divya Madhuri Kallam","doi":"10.1016/j.jchromb.2025.124530","DOIUrl":"10.1016/j.jchromb.2025.124530","url":null,"abstract":"<div><div>The FDA authorized Encorafenib on April 8, 2020, taken along with cetuximab to treat patients (adults) with advanced metastatic colo-rectal cancers, who have a mutation of BRAF-V600E and have previously undergone therapy. No documented techniques for the simultaneous quantitation of Encorafenib and Cetuximab using LC-MS/MS was available, so a reliable, fast, and unique method was developed and validated. Method development involved optimizing chromatographic and mass spectrometric conditions to achieve high sensitivity and specificity. The method was validated per FDA guidelines, evaluating parameters such as linearity, precision, accuracy, recovery, and stability under various conditions. Mass ion pairs were tracked using multiple reaction monitoring (MRM) in positive polarity mode and the precursor to daughter ion transition <em>m</em>/<em>z</em> values for Encorafenib, Cetuximab(peptide), and Tofacitinib (internal reference) are 540.15 → 369.85, 643.34 → 653.31, and 313.17 → 221.05, respectively. The calibration curves demonstrated excellent linearity over 3.75–150 ng/mL ranges for Encorafenib and 0.25–10 ng/mL for Cetuximab. The MS² system offers a significant advantage with its ability to specifically target ions of interest. The technique was effectively employed to quantitate the levels of analytes, key pharmacokinetic parameters were assessed in rats following single-dose administration. Encorafenib exhibited a Cmax of 72.543 ng/mL, Tmax of 2 h, and T1/2 of 20 h, whereas Cetuximab showed a Cmax of 4.982 ng/mL, Tmax of 1 h, and T1/2 of 10 h. Stability studies confirmed the analytes' robustness under various conditions. These findings highlight the method's utility for accurate monitoring of pharmacokinetic parameters, essential for therapeutic drug monitoring in biological samples. It tracks drug levels drugs from administration to several hours post-dose at set intervals, enabling the evaluation of metabolism, excretion, and protein binding, which aid in the creation of treatment plans. It also facilitates routine monitoring of selected medications in clinical trials.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1255 ","pages":"Article 124530"},"PeriodicalIF":2.8,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143471331","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":"Imatinib and norimatinib therapeutic monitoring using dried blood spots: Analytical and clinical validation, and performance comparison of volumetric collection devices","authors":"Marco Orleni ,&nbsp;Sara Gagno ,&nbsp;Eleonora Cecchin ,&nbsp;Marcella Montico ,&nbsp;Angela Buonadonna ,&nbsp;Arianna Fumagalli ,&nbsp;Michela Guardascione ,&nbsp;Fabio Puglisi ,&nbsp;Giuseppe Toffoli ,&nbsp;Bianca Posocco ,&nbsp;Erika Cecchin","doi":"10.1016/j.jchromb.2025.124526","DOIUrl":"10.1016/j.jchromb.2025.124526","url":null,"abstract":"<div><div>Therapeutic drug monitoring during imatinib treatment is recommended to optimize patient clinical outcomes. This study aimed to develop a novel LC-MS/MS method to quantitate imatinib and its active metabolite <em>N</em>-desmethyl-imatinib, in volumetric dried blood spots (DBS) using the HemaXis DB10 and Capitainer B devices. Chromatographic separation was achieved using an XTerra MS C18 column and detection occurred with a SCIEX 4000QTrap tandem mass spectrometer using electrospray positive-mode ionization. Analytical validation was successfully performed adhering to the latest guidelines. The assay was linear over the range 240–6000 ng/mL for imatinib and 48–1200 ng/mL for its metabolite, accurate (89 %–113 %) and precise (≤17 % imprecision) across a hematocrit range of 22–55 % for both devices. Recovery ranged from 84 % to 92 %, with no influence of matrix components. Stability was confirmed after at least 43 days in desiccator conditions (20 °C, ≤35 % humidity), and in conditions that mimed home-sampling. Clinical validation, conducted on 52 paired DBS and plasma samples from 28 patients, revealed that the DBS-to-plasma ratio can be used to convert DBS measurements into plasma concentrations. Bland-Altman and Passing-Bablok analyses indicated strong agreement between the estimated and actual plasma concentrations for both imatinib and its metabolite across both devices. The conversion method was further tested on an additional set of 25 to 31 samples, with 80 to 97 % of the samples falling within ±20 % difference. This study proved that DBS collected using either HemaXis DB10 or Capitainer B devices can be reliably implemented as an alternative to plasma for therapeutic drug monitoring during imatinib therapy.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1255 ","pages":"Article 124526"},"PeriodicalIF":2.8,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143463682","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":"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}