Journal of Chromatography B最新文献

{"title":"Tandem mass tag-based proteomics analysis to reveal growth stage-dependent pathways in Dendrobium nobile Lindl","authors":"Wanhui Chen ,&nbsp;Cong Xie ,&nbsp;Wenying He","doi":"10.1016/j.jchromb.2025.124647","DOIUrl":"10.1016/j.jchromb.2025.124647","url":null,"abstract":"<div><h3>Objective</h3><div>This study aimed to analyze differentially proteins at various growth stages of <em>Dendrobium nobile Lindl</em> and attempt to find some possible functional proteins related to plant growth or pharmacological activity.</div></div><div><h3>Methods</h3><div>The samples were grouped into two categories based on their growth periods as 2–3 years (S1) and 7–8 years (S2), respectively. Tandem Mass Tag(TMT) labeling-based quantitative proteomics combined LC-MS/MS was applied to identify differential protein expression at various growth stages of <em>Dendrobium nobile</em> Lindl<em>.</em> Those proteins were analyzed by various bioinformatics methods. The activities of several antioxidant enzymes including superoxide dismutase(SOD), peroxidase (POD) and catalase (CAT) in the plant were specifically examined using commercially available assay kits.</div></div><div><h3>Results</h3><div>A total of 6032 proteins were identified with 283 differential proteins related to 168 up-regulated and 115 down-regulated. KEGG pathway enrichment analysis revealed that proteins involved in heat shock response, phenylpropanoid biosynthesis, and antioxidant activity showed significant changes across the growth stages. Specifically, a decrease in small heat shock proteins (sHSPs) was observed in older plants, potentially reducing their stress resilience, while proteins involved in lignin biosynthesis, such as SOD, CAT, and POD, were upregulated, suggesting improved stress response.</div></div><div><h3>Conclusion</h3><div>There are 283 differential proteins with diversiform function between different growth stages. Some upregulated antioxidant enzymes contribute to Dendrobium's ability to combat oxidative stress in older plants. These findings provide insights into how protein expression varies with growth stage, offering scientific support for determining optimal harvesting periods or pharmacological mechanism for <em>Dendrobium nobile</em> Lindl<em>.</em></div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1261 ","pages":"Article 124647"},"PeriodicalIF":2.8,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144084672","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":"Optimized plasmid extraction through controlled temperature, prolonged alkaline lysis, and gentle mixing","authors":"Ruijie Hao ,&nbsp;Xingyu Li ,&nbsp;Ning Wang ,&nbsp;Mengzhen Zhang ,&nbsp;Min Li ,&nbsp;Yongjie Xu ,&nbsp;Pengpeng Zhang ,&nbsp;Hanling Zhang ,&nbsp;Zihan Ma ,&nbsp;Bijie Jiang ,&nbsp;Xuefeng Wei ,&nbsp;Wei Xu","doi":"10.1016/j.jchromb.2025.124646","DOIUrl":"10.1016/j.jchromb.2025.124646","url":null,"abstract":"<div><div>Alkaline lysis serves as an effective technique for plasmid extraction. However, its widespread adoption is significantly impeded by inconsistencies in key technical parameters across different documents. To address these issues, we conducted a rheological behavior analysis to understand the pattern of cell lysis and performed a series of plasmid extraction experiments to determine the optimal technical parameters. The result showed that gentle mixing (inverting tube five times, I5)), extended lysis time (10 min), and elevated lysis temperature (25 °C), each produced the best plasmid preparations, based on plasmid yield, quality, and performance in downstream molecular biology assays. We also discovered that bacterial density influenced cell lysis process. Extending the alkaline lysis duration neither induced resistance to restriction endonuclease (RE) digestion of plasmids nor significantly compromised the integrity of plasmids and host genome; instead, it enhanced the plasmid release. High-frequency and small-amplitude mixing tended to generate more genomic fragments, whereas low-frequency and large-amplitude mixing was likely to result in a higher occurrence of open circular (OC) plasmids. Bacterial lysis at low temperatures (4 °C) could lead to genomic contamination and reduce lysis efficiency. Overall, these findings underscored the importance of technical details and provided guidance for future plasmid extractions.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1262 ","pages":"Article 124646"},"PeriodicalIF":2.8,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144105689","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":"Rapid separation and quantification of imidazole dipeptides in meats using a PBr column packed with 3-(pentabromobenzyloxy)propyl group modified silica gel","authors":"Makoto Ozaki ,&nbsp;Tomomi Nakade ,&nbsp;Yasunari Yamada ,&nbsp;Tsunehisa Hirose ,&nbsp;Motoshi Shimotsuma ,&nbsp;Shozo Tomonaga","doi":"10.1016/j.jchromb.2025.124660","DOIUrl":"10.1016/j.jchromb.2025.124660","url":null,"abstract":"<div><div>Meats are rich in imidazole dipeptides (IDPs) such as carnosine, anserine, and balenine, known for their antioxidant and antifatigue properties. The concentrations and types of these IDPs vary significantly among different animal species, necessitating a quantitative method for the precise measurement of individual IDPs. Simultaneous analysis of multiple compounds is typically conducted using reversed-phase high-performance liquid chromatography (RP–HPLC). However, C₁₈ columns, which are commonly employed in RP–HPLC, fail to adequately retain highly hydrophilic IDPs, making separation and quantification challenging. Previously, we developed a PBr column packed with 3-(pentabromobenzyloxy)propyl group modified silica gel, which effectively retains various highly hydrophilic compounds in RP mode. In this study, we established a method for the rapid separation and quantification of IDPs within 10 min under simplified conditions (isocratic mode) using a single quadrupole liquid chromatography–mass spectrometer (LC–MS) equipped with a PBr column, without the need for derivatization. Linear calibration curves for each IDP were generated using glycyl-L-leucine as the internal standard, with the desolvation temperature of the MS instrument set at 500 °C. The proposed method achieved extraction and recovery rates of IDPs ranging from 100.0 % to 113.5 % at three spiking levels, with no carryover observed, even in samples with high concentrations. Additionally, matrix effects ranged from 95.5 % to 109.6 %, with negligible ion suppression and enhancement effects. Furthermore, the method enabled accurate analysis of IDPs with a relative standard deviation of &lt;15 % in meats from various animal species, including chicken, pork, beef, lamb, mutton, deer, horse, and kangaroo.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1262 ","pages":"Article 124660"},"PeriodicalIF":2.8,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144115950","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":"Quantitative GC-NICI-MS and GC-NICI-MS/MS analysis by using isotopologs as internal standards: A tutorial review","authors":"Dimitrios Tsikas","doi":"10.1016/j.jchromb.2025.124648","DOIUrl":"10.1016/j.jchromb.2025.124648","url":null,"abstract":"<div><div>The use of analytes labelled with stable-isotopes of ²H, ¹³C, ¹⁵N, ¹⁷O and/or ¹⁸O, i.e., the isotopologs, as internal standards is unique and considered the <em>Golden Standard</em> is quantitative analyses based on mass spectrometry. However, the handling with isotopologs deserves a great extent of care and attentiveness from the very begin of the analytical process. Many issues need to be considered in order to create an analytical method that generates close-to-reality concentrations of a certain analyte or of group of analytes of the same or different chemical classes in complex biological samples. They including isotopic purity, stability through the entire analytical process including sampling, derivatization, ionization and collision-induced dissociation (CID). The present work deals specifically with the use of isotopologs as internal standards for the quantitative analysis of endogenous and exogenous substances in plasma, serum and urine samples. It focuses on GC–MS and GC–MS/MS, negative-ion chemical ionization (NICI) with methane as the reagent gas, and selected-ion monitoring (SIM) or selected-reaction monitoring (SRM) using argon as the collision gas. Special attention has been paid to purity of isotopologs, cross-contribution of analyte-internal standard, and stability of isotopologs during NICI and CID, to linearity of analyte-isotope detector response upon the analyte concentration. This tutorial review re-examines and discusses exemplarily previously reported validated GC–MS and GC–MS/MS methods, and gives recommendations regarding the handling with stable-isotope labelled analogs in quantitative analyses.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1262 ","pages":"Article 124648"},"PeriodicalIF":2.8,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144105688","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":"Quantitative analysis of DNDI-6174 using UPLC-MS/MS: A preclinical target site pharmacokinetic study","authors":"Wietse M. Schouten ,&nbsp;Katrien Van Bocxlaer ,&nbsp;Hilde Rosing ,&nbsp;Alwin D.R. Huitema ,&nbsp;Jos H. Beijnen ,&nbsp;Jadel M. Kratz ,&nbsp;Charles E. Mowbray ,&nbsp;Thomas P.C. Dorlo","doi":"10.1016/j.jchromb.2025.124652","DOIUrl":"10.1016/j.jchromb.2025.124652","url":null,"abstract":"<div><div>Leishmaniasis is a neglected parasitic infection that continues to pose a significant global health challenge, with currently limited effective treatment options. DNDI-6174 is a novel orally-active, investigational drug with antileishmanial properties. Herein, a novel ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) method was developed and validated to quantify DNDI-6174 in relevant murine biomatrices, <em>i.e.</em>, K₂EDTA plasma and enzymatically-homogenized skin, spleen and liver to support the translational pharmacokinetic-pharmacodynamic model-informed drug development. The chromatographic system consisted of a gradient elution on a standard C₁₈ column connected to a triple quadrupole MS, operating in positive ionization mode. Pre-processing of murine tissues with collagenase A led to a superior homogenization and analyte extraction compared to mechanical disruption. Human K₂EDTA plasma served as a surrogate matrix, enabling accurate (bias between −12.0 % and 9.8 %) and precise (relative standard deviation (RSD) ≤ 12.5 %) quantification of DNDI-6174 in the various murine biomatrices. Sample processing with <em>tert</em>-methylbutyl ether resulted in a reproducible recovery between 70.0 % and 93.8 % (RSD ≤ 4.0 %) with an absolute matrix factor between 0.89 and 1.00 for all biomatrices. DNDI-6174 was stable under various conditions, including under tissue homogenization conditions, in all biomatrices investigated. This method was successfully applied in a translational study using a murine cutaneous leishmaniasis skin infection model to assess the target site pharmacokinetics of DNDI-6174, supporting its development as clinical candidate.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1262 ","pages":"Article 124652"},"PeriodicalIF":2.8,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144124799","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":"Quantification of sulbactam and durlobactam in saline and human plasma via ultra-performance liquid chromatography tandem mass spectrometry","authors":"Hanna F. Roenfanz,&nbsp;David P. Nicolau,&nbsp;Joseph L. Kuti","doi":"10.1016/j.jchromb.2025.124654","DOIUrl":"10.1016/j.jchromb.2025.124654","url":null,"abstract":"<div><div>Sulbactam-durlobactam is a combination β-lactam/β-lactamase inhibitor antibiotic approved in the United States for the treatment of adult patients with hospital-acquired and ventilator-associated bacterial pneumonia due to susceptible isolates of <em>Acinetobacter baumannii-calcoaceticus</em> complex. A liquid chromatography tandem mass spectrometry method for the quantification of sulbactam and durlobactam in human plasma and aqueous matrices has been developed and validated. The standard curves for each drug were linear over the range 0.5 to 50 μg/mL and use the isotopic analogs sulbactam-d₅ and [¹³C₂, ¹⁵N₂]-durlobactam as internal standards for their respective analytes. This simple, reproducible method for the determination of sulbactam and durlobactam concentrations was developed with the intent to conduct future pharmacokinetic analyses and to guide clinical laboratories in the development of a therapeutic drug monitoring assay.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1261 ","pages":"Article 124654"},"PeriodicalIF":2.8,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144071200","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":"Immobilization of albumin binding domain (ABD) on Sepharose 4B and magnetic particle for efficient single-step purification of human serum albumin","authors":"Maryam Nazari ,&nbsp;Rahman Emamzadeh ,&nbsp;Nastaran Masoudi-Khoram ,&nbsp;Mahboobeh Nazari","doi":"10.1016/j.jchromb.2025.124655","DOIUrl":"10.1016/j.jchromb.2025.124655","url":null,"abstract":"<div><div>Human serum albumin (HSA) is an important protein in plasma with various biological functions in the human body. Due to its unique features in the binding and transfer of ligands and pharmaceutical molecules, HSA is extensively used in therapeutics and pharmaceutical approaches. Commercial albumin is produced by a multi-step process of plasma fractionation. However, this traditional method has some limitations such as risk of contamination, low quality, and quantity of the purified final protein. In this study, we developed two affinity chromatography platforms for the purification of human serum albumin. The recombinant albumin-binding domain (ABD) was expressed and purified using molecular biology techniques. Two types of commercial beads—Cyanogen bromide-activated Sepharose 4B and amine-functionalized magnetic particles—were then functionalized with the recombinant ABD. Protein purification using chromatography columns demonstrated that HSA can be purified to 95 % purity in a single step. Circular dichroism (CD) spectroscopy revealed structural similarities in HSA purified through affinity chromatography and fractionation using the Cohen method. Furthermore, the study of aspirin binding to HSA demonstrated that proteins purified via affinity chromatography and those fractionated by the Cohen method exhibited identical drug-binding affinities. The results of this study may have important implications for the clinical purification of human serum albumin.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1261 ","pages":"Article 124655"},"PeriodicalIF":2.8,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144098873","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 simple surrogate approach for the quantitation of C4 (7α-hydroxy-4-cholesten-3-one) in human serum via LC-MS/MS and its application in a clinical study","authors":"Feng Yin ,&nbsp;Mani Deepika Vakkalanka ,&nbsp;Walter Wiley ,&nbsp;M. Shane Woolf ,&nbsp;Yousef Basir ,&nbsp;Kumar Shah ,&nbsp;Aaron M. Wheeler ,&nbsp;Moucun Yuan ,&nbsp;William R. Mylott Jr. ,&nbsp;Mike Baratta","doi":"10.1016/j.jchromb.2025.124651","DOIUrl":"10.1016/j.jchromb.2025.124651","url":null,"abstract":"<div><div>We present a validated LC-MS/MS assay for the quantitation of 7α-hydroxy-4-cholesten-3-one (C4), a key intermediate in the bile acid synthesis pathway from cholesterol, in human serum. A surrogate matrix approach was employed to overcome the challenges posed by the endogenous C4 levels in the biological matrix. Human serum samples were spiked with stable isotope labeled internal standard (SIL-IS), processed using supported liquid extraction (SLE), and analyzed by LC-MS/MS. Parallelism was successfully demonstrated between human serum (authentic matrix) and 5 % bovine serum albumin in phosphate buffered saline containing 0.1 % tween 20 (5 % BSA in PBST) (surrogate matrix). The assay's linear analytical range was established from 0.200 to 200 ng/mL. This validated LC-MS/MS method exhibited excellent accuracy and precision. The overall accuracy was between 97.9 % and 101 % with %CV less than 4.0 % for C4 in human serum. C4 was found to be stable in human serum for up to 24.7 h at room temperature, up to 34 days when stored at −25 °C or − 80 °C, and after five freeze/thaw cycles. The assay has been successfully applied to human serum samples to support a clinical study.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1261 ","pages":"Article 124651"},"PeriodicalIF":2.8,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144071196","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 magnetic poly (methacrylic acid-co-ethylene glycol dimethacrylate) copolymer for extraction of anticonvulsants from saliva followed by high performance liquid chromatography analysis","authors":"Camila Gabriela Carrara ,&nbsp;Diailison Teixeira de Carvalho ,&nbsp;Mariana Azevedo Rosa ,&nbsp;Luiz Fernando Gorup ,&nbsp;Maria Betânia de Freitas Marques ,&nbsp;Eduardo Costa Figueiredo","doi":"10.1016/j.jchromb.2025.124635","DOIUrl":"10.1016/j.jchromb.2025.124635","url":null,"abstract":"<div><div>Neurological diseases like epilepsy are commonly treated with anticonvulsants, and the dosage control of these medications has been carried out with therapeutic biomonitoring, mainly in blood. Saliva is a less invasive sample, and its drug concentration is proportional to the blood fraction concentration not bound to the proteins, making it an important alternative to blood. Therefore, this study describes the synthesis of a magnetic poly (methacrylic acid-<em>co</em>-ethylene glycol dimethacrylate) copolymer (M-CP) and its use in magnetic dispersive solid phase extraction of primidone (PMR), phenobarbital (PB), phenytoin (PHT), and carbamazepine (CB) from saliva, followed by their determination by high performance liquid-chromatography. M-CP was synthesized in 4 steps and the material was characterized by infrared spectroscopy, scanning electron microscopy, zeta potential, and thermal analysis. The sample preparation protocol was optimized by central composite rotatable design and the optimal conditions were obtained for pH 5.78, 12.8 mg of M-CP, and 2.18 mL of diluted saliva. In the adsorption studies, the Avrami and Jovanovic models demonstrated the best fit for describing the adsorption kinetics and isotherm, respectively. The equilibrium time was reached in 60 min, and 10.96 mg g⁻¹ was the maximum adsorption capacity. The developed method was linear (R² &gt; 0.99) in the range of 2 to 30 mg L⁻¹ for PMR and PB, 1 to 20 mg L⁻¹ for PHT, and 1 to 30 mg L⁻¹ for CBZ. Precisions were obtained in the range of 1.35 to 19.44 %, and accuracy from −14.92 % to 18.83 %. The limits of detection ranged from 0.48 to 1.30 mg L⁻¹, while the limits of quantification were lower than the therapeutic levels of each drug, being suitable for the method application. The developed method is simple, fast, and requires few amounts of saliva, sorbent, and solvents, being a good alternative for the therapeutic biomonitoring of anticonvulsants in saliva samples.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1261 ","pages":"Article 124635"},"PeriodicalIF":2.8,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144071199","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 tacrolimus in whole blood of pediatric liver transplant patients by UPLC–MS/MS: Informed its individualized dose","authors":"Saisai Ling ,&nbsp;Ying Jin ,&nbsp;Cuiyao He ,&nbsp;Lisha Fu ,&nbsp;Yuhua Deng ,&nbsp;Xiaohui Qi ,&nbsp;Zhenglei Wang ,&nbsp;Yiying Wu","doi":"10.1016/j.jchromb.2025.124645","DOIUrl":"10.1016/j.jchromb.2025.124645","url":null,"abstract":"<div><div>Tacrolimus is a potent macrolide immunosuppressant widely used in pediatric liver transplant patients. However, its narrow therapeutic window and significant inter-individual pharmacokinetic differences result in a poor correlation between blood concentrations and administered doses. To support its individualized dose, a sensitive, fast and robust ultra-high performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was developed to measure tacrolimus concentrations in whole blood of pediatric liver transplant patients. The method employed acetonitrile for protein precipitation, and sample separation was achieved using an Acquity UPLC CSH C₁₈ column (2.1 × 50 mm, 1.7 μm) with gradient elution. The mobile phase consisted of ammonium solution–water (0.5:1000, <em>v</em>/v) and methanol. The method demonstrated good linearity within a concentration range of 0.20–50.00 ng/mL. The intra- and inter-day precisions of tacrolimus in whole blood were less than 13.5 %, with the accuracy ranged from −7.2 % to 13.0 %. Selectivity, carryover, matrix effects, recovery, dilution reliability, and stability all met the standards set by relevant guidelines. The established UPLC–MS/MS method was successfully applied to measure the concentration of tacrolimus in the whole blood of pediatric liver transplant patients and support its individualized dose. Under the rapid UPLC–MS/MS method, 25 (65.8 %) patients required a dose increase, 3 (7.9 %) patients needed a dose reduction, and 10 (26.3 %) patients maintained their original dosage without adjustment.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1261 ","pages":"Article 124645"},"PeriodicalIF":2.8,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143935284","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}