Journal of Chromatography B最新文献

{"title":"Effect of furmonertinib on the pharmacokinetics of rivaroxaban or apixaban in vivo.","authors":"Zhi Wang, Zefang Yu, Lingzhi Fang, Jing An, Chaojun Xue, Xin Zhou, Xiao Li, Ying Li, Zhanjun Dong","doi":"10.1016/j.jchromb.2024.124425","DOIUrl":"10.1016/j.jchromb.2024.124425","url":null,"abstract":"<p><p>Furmonertinib, a third generation epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs), is used for non-small cell lung cancer (NSCLC). Rivaroxaban and apixaban are direct oral anticoagulants (DOACs) used for venous thromboembolism (VTE), which is a frequent comorbid with NSCLC. They are substrates of CYP3A4, P-gp and BCRP, whereas furmonertinib is an inhibitor of P-gp and BCRP. This study aimed to disclose the extent of effect of furmonertinib on the pharmacokinetics of rivaroxaban or apixaban. Rats were divided into four groups (n = 6) that received rivaroxaban (group 1), furmonertinib and rivaroxaban (group 2), apixaban (group 3), furmonertinib and apixaban (group 4). The concentrations of drugs were measured by an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Furmonertinib increased the C_max and AUC_0-t of rivaroxaban by 1.66 and 2.07-fold, whereas decreased the CL_z/F by 1.70-fold and V_z/F 1.27-fold. Furthermore, furmonertinib caused similar changes in apixaban pharmacokinetics. The pharmacokinetic results suggest that it is essential to alert the effect of furmonertinib on the pharmacokinetics of rivaroxaban or apixaban in clinical practice.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"124425"},"PeriodicalIF":2.8,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142826959","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 quantification method for direct oral anticoagulants from capillary blood using volumetric absorptive microsampling and online SPE-LC-MS.","authors":"Patrick Opitz, Isabel Waltering, Georg Hempel","doi":"10.1016/j.jchromb.2024.124423","DOIUrl":"10.1016/j.jchromb.2024.124423","url":null,"abstract":"<p><p>The number of prescriptions for new direct oral anticoagulants (DOACs) apixaban, edoxaban, rivaroxaban and dabigatran has increased exponentially in recent years, increasingly replacing the old gold standard, vitamin-K-antagonists. Due to their wide therapeutic range, therapeutic drug monitoring (TDM) is not required, although it has been proven that this could significantly reduce side effects. In order to develop a cost-efficient and simple method for the simultaneous detection of the DOACs and phenprocoumon, a new technology for sample preparation from capillary blood in the ambulant sector named VAMS® was integrated and an LC-MS detector with on-line solid phase extraction (SPE) applying a Turboflow HTLC Cyclone^TM 1.0x50 mm column was used. The mobile phase consisted of methanol with water (3/97 v/v) and 0.1 % ammonia solution with a flow rate of 2.5 mL/min. For the chromatographic separation, a Phenomenex LTD Kinetex 2.6 µm C18 100 Å, 100x3.0 mm column with a flow rate of 0.3 mL/min in gradient mode was utilized. The mobile phase consisted of acetonitrile, water and formic acid (A: 10:90:0.1 v/v and B: 95:05:0.1 v/v). The method was fully validated in the therapeutic range of the substances according to current guidelines. The LLOQ ranged from 3.5 µg/L for rivaroxaban to 88 µg/L for phenprocoumon and the intra-day and inter-day precision was less than 13 % and 12 %, while the accuracy was within a range of 85.7-113 % and 88.7-106 %, respectively. Samples could be stored in the Mitra® devices for at least seven days at room temperature except of dabigatran. Because the Mitras® were used, exactly 10 µL of blood could be drawn and no significant haematocrit effect was observed. A reliable, simple and cost-effective extraction and analysis LC-MS method could be developed and validated. This method is therefore applicable in ambulatory care.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"124423"},"PeriodicalIF":2.8,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142833561","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":"Study on the protective mechanism of Xuemaitong Capsule against acute myocardial ischemia rat based on network pharmacology and metabolomics.","authors":"Jialu Zou, Shizhong Zhang, Xiaohong Zhang, Lijuan Xiong, Xuan Chen, Yanmei He, Cancan Duan, Jianyong Zhang","doi":"10.1016/j.jchromb.2024.124373","DOIUrl":"10.1016/j.jchromb.2024.124373","url":null,"abstract":"<p><strong>Background: </strong>Xuemaitong Capsule (XMT) is a widely recognized traditional Miao medicine extensively utilized in Chinese clinical settings. Previous studies have demonstrated XMT protective effects against acute myocardial ischemia (AMI). However, the mechanism by which XMT provides protection to AMI rats is yet to be fully understood.</p><p><strong>Aim of the study: </strong>The purpose of this study was to investigate the protective mechanism of XMT on AMI rats through network pharmacology, traditional pharmacodynamics and metabolomics.</p><p><strong>Material and methods: </strong>The components and potential targets of XMT were identified through the application of traditional Chinese medicine system pharmacology and traditional Chinese medicine molecular mechanism bioinformatics analysis tools. We constructed herb-composition-target networks and analyzed protein-protein interaction (PPI) networks. The potential mechanism was explored by pathway enrichment analysis. Subsequently, the AMI model was constructed by ligation of the anterior descending branch of the left coronary artery, and XMT protective effects on AMI rats were evaluated by analyzing the myocardial enzyme profiles, electrocardiograms(ECG), Triphenyltetrazolium chloride(TTC) staining, and Hematoxylin-Eosin (HE) staining in AMI rats. Metabolomics based on UHPLC-Q-Exactive Orbitrap MS was used to observe the protective effect of XMT on the serum metabolic profile of AMI, and multivariate statistical analysis further revealed the differential patterns of metabolites after XMT treatment. Finally, integrated pathway analysis was carried out to reveal the biological metabolic mechanism.</p><p><strong>Results: </strong>A total of 392 active components of XMT acted with 624 targets for treating AMI. Pathway enrichment analysis revealed that XMT could treat AMI through TNF, MAPK and PI3K-Akt signaling pathways. Further, XMT could effectively prevent ST-segment elevation in the ECG, reduce the size of myocardial infarction, decrease cardiac weight index and cardiac enzyme levels, and mitigate histological damage in the hearts of AMI rats. In addition, XMT callback 117 metabolites and four metabolic pathways, including taurine and hypotaurine metabolism, phenylalanine metabolism, pyrimidine metabolism and retinol metabolism. Through integrating network pharmacology and metabolomics, we explored the biological mechanism by which XMT treats AMI. It was speculated that the mechanism of XMT is to regulate TNF signaling, PI3K-Akt pathway and MAPK signaling pathway, and participate in cell apoptosis, oxidative stress, immune and inflammatory reaction and other biological processes.</p><p><strong>Conclusion: </strong>XMT plays a protective role in AMI rats by regulating multiple metabolic biomarkers, multiple targets and pathways. Therefore, XMT may provide a potential strategy for the treatment of AMI.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"124373"},"PeriodicalIF":2.8,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142790813","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":"Non-targeted screening of illegal added hypoglycemic drugs and their derivatives in functional foods using characteristic fragment ions scanning based on liquid chromatography-high-resolution mass spectrometry.","authors":"Yajie Liu, Feng Feng, Xiujuan Wang, Rongzhu Du, Xuesong Feng, Feng Zhang","doi":"10.1016/j.jchromb.2024.124432","DOIUrl":"10.1016/j.jchromb.2024.124432","url":null,"abstract":"<p><p>Functional foods that are illegally adulterated with chemical hypoglycemic drugs or their derivatives pose serious risks to human health. However, the detection of these compounds is a challenge due to the unknown nature of their structures, particularly as many of these compounds are newly synthesized and lack standardized references. In this study, we developed a non-targeted screening strategy for detecting illegally added hypoglycemic drugs and their derivatives in oral solution and hard capsule functional foods. This strategy utilizes a self-established characteristic fragment ions mass spectrum database based on ultrahigh-performance liquid chromatography-quadrupole/orbitrap high-resolution mass spectrometry. Nine characteristic fragment ions were identified by analyzing the mass spectrometric fragmentation patterns of hypoglycemic drugs. Meanwhile, the precursor and fragment ions of 38 known hypoglycemic drugs were collected and incorporated into the mass database. The detection limits of hypoglycemic drugs in oral solution and hard capsule samples were 0.01-10 μg/L and 0.01-50 μg/kg, respectively. Applying the non-targeted method to 20 batches of suspicious samples, we found that 3 batches of samples contained illegal added drugs. Furthermore, we identified a previously unrecorded hypoglycemic drug not present in the mass database. These results indicate that the developed strategy is a powerful tool for the rapid and highly sensitive identification of both known and unknown hypoglycemic drugs, as well as their derivatives in functional foods.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"124432"},"PeriodicalIF":2.8,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142870648","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":"Xenobiotics recovery from plasma using solid phase extraction with C-18 sorbent - The reasons of literature discrepancies.","authors":"Rafal Typek, Michal P Dybowski, Michal Rombel, Piotr Holowinski, Andrzej L Dawidowicz","doi":"10.1016/j.jchromb.2024.124433","DOIUrl":"10.1016/j.jchromb.2024.124433","url":null,"abstract":"<p><p>Solid phase extraction (SPE) is one of the most popular methods of preparing plasma samples before determining the xenobiotics they contain. The present paper shows that the recovery degree of xenobiotics from plasma samples using SPE with C18 sorbent strongly depends on their storage time and temperature. While xenobiotics can be isolated and recovered in 100 % from fresh plasma samples under optimal conditions of the SPE procedure, their SPE recovery degree from stored plasma is lower. It diminishes with the time increase and temperature reduction of plasma storage. Moreover, a part of xenobiotic in stored plasma samples is not sorbed on SPE-C18 column at all and leaves it together with the waste during loading the column with the examined sample. According to the NMR data, the main reason of the presence of xenobiotics in the waste of the SPE column during its loading are structural-chemical changes occurring in plasma during its storage, leading to the formation of some complex(es) of hydrophilic character with xenobiotic. Another reason for lower SPE recovery degree of xenobiotic from stored plasma is the formation of plasma sediment, which binds/occludes xenobiotic. The presented results expand the current knowledge on why the SPE recovery degree of xenobiotics from plasma reported in the literature may differ. They are important for proper calibration of the chromatographic system in the analysis of xenobiotics in plasma and for achieving high accuracy of the analytical procedure involving SPE in xenobiotic estimation.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"124433"},"PeriodicalIF":2.8,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142870653","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 analysis of active compounds targeting tropomyosin-related kinase A via constructed lipid raft @capillary monolith affinity chromatography.","authors":"Hongbei Liu, Qiumin Xu, Michael Adu-Frimpong, Yuchu Chen, Ran Li, Fei Xu, Xia Cao, Shanshan Tong","doi":"10.1016/j.jchromb.2024.124429","DOIUrl":"10.1016/j.jchromb.2024.124429","url":null,"abstract":"<p><p>In order to enrich the selection of biological ligands, realize the miniaturization analysis, and broaden the application of monolith materials for active ingredients screening and separating, we sough to construct a lipid raft @capillary monolith microcolumn affinity chromatography model. Single factor experiments and various characterization methods, including scanning electron microscopy (SEM) and thermogravimetric analysis, were employed to investigate the polymerization of the monolith column under different material ratios to determine optimal preparation conditions. Subsequently, the lipid raft from U251 cells was integrated with the monolith materials based on epoxy-based covalent crosslinking principle and characterized through SEM and immunofluorescence methods. Afterwards, the retention of positive drug gefitinib, negative drug gemcitabine and four licorice standards solution on the prepared lipid raft monolith microcolumn was then detected via electrochemical detection. The results exhibited that there was no specific adsorption for any active compounds on the blank monolith materials. Significantly, the lipid raft monolith microcolumn packed with TrkA-target proteins could be successfully validated for positive drug gefitinib with a high affinity sorption efficiency of 51.2%. This work expands the range of the utilization of affinity chromatography carriers and the selection of biological ligands, providing a new idea for the screening of active ingredients.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"124429"},"PeriodicalIF":2.8,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142890991","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":"Determination of four forms of plasma thiol amino acids in individuals with chronic kidney disease by UPLC-MS/MS.","authors":"Yuyu Cao, Dayi Xu, Liping Zhang, Kaiyuan Pang, Liyuan Zhang, Xiaobao Wei, Zengxian Sun","doi":"10.1016/j.jchromb.2024.124418","DOIUrl":"10.1016/j.jchromb.2024.124418","url":null,"abstract":"<p><p>The study introduces a robust analytical method based on UPLC-MS/MS for quantifying thiol amino acids, including cysteine (Cys), cysteinylglycine (CG), homocysteine (Hcy), and glutathione (GSH), in their total and total free forms within human plasma. An optimized blank matrix was employed for accurate quantification of endogenous compounds. The method exhibited excellent linearity, precision, accuracy, recovery, and stability, making it highly suitable for plasma analysis. Distinct differences in plasma levels of GSH, Cys, Hcy, and CG across various forms (total, total free, native prototype, and symmetrical oxidation) were observed between healthy individuals and chronic kidney disease (CKD) patients. Comprehensive correlation and receiver operating characteristic (ROC) analyses revealed disrupted thiol amino acid metabolism in CKD, accompanied by heightened oxidative stress. Notably, various forms of Cys and Hcy correlated significantly with renal function markers and demonstrated high diagnostic efficacy in ROC evaluation, with Cys, particularly Cys (F), outperforming others. Hcy further complements Cys in assessing renal function impairment severity.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"124418"},"PeriodicalIF":2.8,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142890986","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":"Chromatographic retention assisted in-silico prediction of anticancer potential of glucocorticoids on cancer cell lines.","authors":"Faten Farouk, Marwa Sharaky, Ehab F Elkady, Rawda M Sayed","doi":"10.1016/j.jchromb.2025.124466","DOIUrl":"https://doi.org/10.1016/j.jchromb.2025.124466","url":null,"abstract":"<p><p>Glucocorticoids (GCs) are hallmarks of anti-inflammatory activity. They are used as adjuvant therapy in oncology medications to alleviate some of the associated side effects. Although recent research has indicated that GCs have favorable anticancer potential, some scientific evidence suggests a pro-proliferation impact of GCs on cancer cells. This may create a scientific confusion on the utility of GCs and the choice of which GC enhances the anticancer potential and mitigates the negative effect. Accurate in-silico prediction and correct ranking of biological activity may be limited by the impact of the physicochemical interaction between small molecules and biological membranes. Chromatographic retention is inherently dependent on the physicochemical properties of the test molecule. It can scale the relative hydrophobicity and tentatively indicate the membrane permeability. In this study, the relationship between the in-silico binding affinity, the chromatographic retention and the in-vitro anticancer activity was investigated. Fifteen GCs were chromatographically separated on an Inertsil® C18 (4.6 * 250 mm; 5 μm) HPLC column. The binding affinity of the test GCs was determined on three receptors involved in cancer cell proliferation (topoisomerase II (TOPII), glucocorticoid receptor (GR) and ATP-binding cassette (ABCG2)). The antiproliferative potential of the test steroids was determined as per their IC₅₀. The correlation between chromatographic retention and the binding affinity to the observed IC₅₀ was investigated by multiple linear regression (MLR). Results revealed that some GCs exhibited a remarkably favorable inhibitory potential against cancer cell lines over normal cell lines. Our data indicated a significant correlation between the retention times of different GCs and the determined binding affinity, especially to the GR (r² = 0.677; p = 0.011) and the estrone sulphate (ESS) binding site of the ABCG2 (r² = 0.643; p = 0.018). Concurrently, the retention times were well correlated to the observed IC₅₀ on the colorectal cancer cell lines (r² = 0.580; p = 0.038) and the hypopharyngeal cell lines (r² = 0.638; p = 0.019). Significant MLR models (n = 4) correlating the retention times of the tested steroids and the binding affinity to the observed IC₅₀ were created. The MLR model correlating the retention times and the ABCG2_ESS binding affinity to the IC₅₀ on lung cancer was the most significant (p = 0.004). The accuracy of the model was 107.12 ± 29.18 % indicating good IC₅₀ prediction abilities. In conclusion, chromatographic retention can be employed as a low-cost and simple auxiliary tool for improving the in-silico prediction of the in-vitro activities of small molecules.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1253 ","pages":"124466"},"PeriodicalIF":2.8,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143035706","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":"Determination of eldecalcitol in human plasma by SIL-IS UPLC-APCI-MS/MS method for pharmacokinetics study.","authors":"Wanyu Wang, Zhichao Yin, Pengfei Du, Jianbang Wu, Minhui Wang, Yaqin Wang, Ping Wu, Xiaocui Xia, Lin Zhang, Yamin Liu, Zhenyue Gao, Jie Shen, Yuanwei Jia","doi":"10.1016/j.jchromb.2025.124468","DOIUrl":"https://doi.org/10.1016/j.jchromb.2025.124468","url":null,"abstract":"<p><p>Eldecalcitol is a novel analog of 1α,25-dihydroxyvitamin D₃ [1,25(OH)₂D₃] for the treatment of patients with osteoporosis. A highly sensitive and specific ultra-performance liquid chromatography coupled to atmospheric pressure chemical ionization tandem mass spectrometry (UPLC-APCI-MS/MS) technique featuring a lower limit of quantitation (LLOQ) as low as 5 pg/ml, has been established and validated for the rapid and accurate quantification of eldecalcitol in human plasma samples. Plasma samples were extracted by solid phase extraction (SPE). Stable isotope-labeled compound eldecalcitol-d6 was used as an internal standard (SIL-IS). The detection process was carried out utilizing Multi-Reaction Monitoring (MRM) mode, employing an atmospheric pressure chemical ionization (APCI) source operating in the positive ion modality. The target fragment ion pairs for eldecalcitol and SIL-IS were identified as m/z 508.6 transitioning to 397.4, and m/z 514.6 transitioning to 403.3, respectively. This method was validated regarding selectivity, LLOQ, linearity, accuracy and precision, recovery, matrix effects, dilution reliability, stability, carryover test, and incurred sample reanalysis (ISR). This methodology had been successfully employed to assess the pharmacokinetics (PK) profile of eldecalcitol in a clinical investigation.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1252 ","pages":"124468"},"PeriodicalIF":2.8,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142997686","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 UHPLC-MS/MS method for the quantitative analysis of trans-ISRIB in human plasma.","authors":"Weizhuan He, Akshay Suresh Patil, Yan Xu","doi":"10.1016/j.jchromb.2025.124469","DOIUrl":"https://doi.org/10.1016/j.jchromb.2025.124469","url":null,"abstract":"<p><p>The integrated stress response (ISR) is a cellular defense mechanism activated under stress conditions. When the ISR is activated, it slows the production of proteins, the building blocks that cells need to function. Trans-integrated stress response inhibitor (trans-ISRIB) is a compound that can reverse the effects of ISR activation, showing promise for treating neurodegenerative diseases. The preclinical and clinical evaluation of trans-ISRIB necessitates a reliable analytical method. This study presents the development and validation of an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the quantitative analysis of trans-ISRIB in human plasma, conforming to the U.S. FDA's guidelines for bioanalytical method validation. The method developed utilizes a liquid-liquid extraction procedure to prepare plasma samples with a spiked internal standard (IS). The extracts containing trans-ISRIB and the IS were dried under nitrogen, reconstituted in the mobile phase, and separated on a Waters XSelect HSS T3 column under isocratic conditions with a mobile phase containing 0.1 % acetic acid in 70 % methanol aqueous solution at a flow rate of 0.500 mL/min. Detection and quantification were accomplished using a positive electrospray ionization tandem mass spectrometer (ESI⁺-MS/MS) operated in multiple-reaction-monitoring (MRM) mode. The method demonstrated a linear calibration range for trans-ISRIB concentrations from 0.500 to 1.00 x 10³ nM, with high specificity, precision, accuracy, and recovery. This method addresses a significant analytical gap, offering a robust tool for quantifying trans-ISRIB in human plasma. Chemical compounds studied in this article: 2-(4-chlorophenoxy)-N-[4-[[2-(4-chlorophenoxy)acetyl]amino]cyclohexyl]acetamide (trans-ISRIB) (CAS # 1597403-47-8); 2-(4-chlorophenoxy)-N-(2-{[(4-chlorophenoxy)acetyl]amino}ethyl)acetamide (CAS # 327071-30-7).</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1252 ","pages":"124469"},"PeriodicalIF":2.8,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142997689","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}