Journal of Chromatography B最新文献_第8页

UPLC-MS/MS-based targeted analysis of human serum revealed that aberrant arachidonic acid metabolism may contribute to the pathogenesis of type 2 diabetes mellitus 基于UPLC-MS/ ms的人血清靶向分析显示，花生四烯酸代谢异常可能与2型糖尿病的发病机制有关

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-01-15 Epub Date: 2025-11-25 DOI: 10.1016/j.jchromb.2025.124871

Peng Jin , Kun Wang , Xiao-wan Hu , Yi-fan Yang , Xian-jie Huang , Yan Du , Dao-quan Tang

The aberrant arachidonic acid (ARA) metabolism may contribute to the pathogenesis of type 2 diabetes mellitus (T2DM), and some metabolites from ARA may have benefits for the differential diagnosis of T2DM. Thus, the aim of this study was to develop an ultra-high performance liquid phase coupled with mass spectrometry (UPLC-MS/MS) for determination of ARA and its 11 metabolites in the serum of patients with T2DM and explore the relation between ARA metabolites and T2DM pathogenesis. A cohort of 66 healthy individuals and 141 patients with T2DM enrolled from Suining People's Hospital Affiliated to Xuzhou Medical University was studied. The UPLC-MS/MS technique was developed and applied to obtain the serum ARA and its 11 metabolite profiles. Multivariate statistical analysis referring to metabolomics were applied for the metabolites data analysis and the differential metabolites screening. The results showed that a total number of 6 differential metabolites were presented between T2DM patients and healthy control subjects. Among these differential metabolites, (±)14(15)-epoxyeicosatrienoic acid (EET) presented the 1.64-fold elevation in T2DM patients versus healthy control subjects, significant correlation with biochemical index [fasting blood glucose (FBG) and glycated hemoglobin (HbA1c)] (P < 0.01), and Receiver Operating Characteristic (ROC) area under the curve (AUC) of 0.6063 with the sensitivity of 26.95% and specificity of 93.94% (95% confidence interval, 0.5280–0.6846) for the diagnosis of T2DM. This study advances previous findings by establishing eicosatetraenoic acids flux imbalance as a hallmark of early T2DM pathogenesis, while providing an analytical blueprint for clinical translation of lipid mediator biomarkers in type 2 diabetic patients.

花生四烯酸（ARA）代谢异常可能与2型糖尿病（T2DM）的发病机制有关，ARA的一些代谢物可能有助于T2DM的鉴别诊断。因此，本研究的目的是建立一种超高效液相联用质谱法（UPLC-MS/MS）检测T2DM患者血清中ARA及其11种代谢物，并探讨ARA代谢物与T2DM发病机制的关系。选取徐州医科大学附属遂宁人民医院66名健康个体和141名T2DM患者作为研究对象。建立并应用UPLC-MS/MS技术获得血清ARA及其11种代谢物谱。代谢物数据分析和差异代谢物筛选采用代谢组学的多变量统计分析。结果显示，T2DM患者与健康对照组之间共存在6种差异代谢物。在这些差异代谢物中，（±）14(15)-环氧二碳三烯酸（EET）在T2DM患者中比健康对照组升高1.64倍，与生化指标[空腹血糖（FBG）和糖化血红蛋白（HbA1c）]有显著相关性（P < 0.01），受试者工作特征（ROC）曲线下面积（AUC）为0.6063，诊断T2DM的敏感性为26.95%，特异性为93.94%（95%可信区间为0.528 ~ 0.6846）。本研究进一步证实了二十碳四烯酸通量失衡是早期T2DM发病机制的标志，同时为2型糖尿病患者脂质介质生物标志物的临床翻译提供了分析蓝图。

{"title":"UPLC-MS/MS-based targeted analysis of human serum revealed that aberrant arachidonic acid metabolism may contribute to the pathogenesis of type 2 diabetes mellitus","authors":"Peng Jin , Kun Wang , Xiao-wan Hu , Yi-fan Yang , Xian-jie Huang , Yan Du , Dao-quan Tang","doi":"10.1016/j.jchromb.2025.124871","DOIUrl":"10.1016/j.jchromb.2025.124871","url":null,"abstract":"<div><div>The aberrant arachidonic acid (ARA) metabolism may contribute to the pathogenesis of type 2 diabetes mellitus (T2DM), and some metabolites from ARA may have benefits for the differential diagnosis of T2DM. Thus, the aim of this study was to develop an ultra-high performance liquid phase coupled with mass spectrometry (UPLC-MS/MS) for determination of ARA and its 11 metabolites in the serum of patients with T2DM and explore the relation between ARA metabolites and T2DM pathogenesis. A cohort of 66 healthy individuals and 141 patients with T2DM enrolled from Suining People's Hospital Affiliated to Xuzhou Medical University was studied. The UPLC-MS/MS technique was developed and applied to obtain the serum ARA and its 11 metabolite profiles. Multivariate statistical analysis referring to metabolomics were applied for the metabolites data analysis and the differential metabolites screening. The results showed that a total number of 6 differential metabolites were presented between T2DM patients and healthy control subjects. Among these differential metabolites, (±)14(15)-epoxyeicosatrienoic acid (EET) presented the 1.64-fold elevation in T2DM patients versus healthy control subjects, significant correlation with biochemical index [fasting blood glucose (FBG) and glycated hemoglobin (HbA1c)] (<em>P</em> < 0.01), and Receiver Operating Characteristic (ROC) area under the curve (AUC) of 0.6063 with the sensitivity of 26.95% and specificity of 93.94% (95% confidence interval, 0.5280–0.6846) for the diagnosis of T2DM. This study advances previous findings by establishing eicosatetraenoic acids flux imbalance as a hallmark of early T2DM pathogenesis, while providing an analytical blueprint for clinical translation of lipid mediator biomarkers in type 2 diabetic patients.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1269 ","pages":"Article 124871"},"PeriodicalIF":2.8,"publicationDate":"2026-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145621746","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}

引用次数: 0

An optimized LC-ESI-MS/MS assay for erythrocyte 6-TG and 6-MMPD: Addressing critical methodological considerations for thiopurine metabolite monitoring 红细胞6-TG和6-MMPD的优化LC-ESI-MS/MS分析：解决硫嘌呤代谢物监测的关键方法学问题。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-01-15 Epub Date: 2025-11-26 DOI: 10.1016/j.jchromb.2025.124872

Ya-Min Chu , Yuan-Yuan Zhang , Ya-Hui Hu , Jie Wang , Hong-Li Guo , Jin Xu , Yong-Jun Fang , Jing Xu , Xuan-Sheng Ding , Feng Chen

Thiopurines, a class of antimetabolites widely used in patients with inflammatory bowel disease (IBD) and acute lymphoblastic leukemia (ALL), are associated with two predominant adverse effects: delayed myelotoxicity and hepatotoxicity. The erythrocyte-based active metabolites 6-thioguanine nucleotides (6-TGN) and 6-methylmercaptopurine nucleotides (6-MMPN) serve as biomarkers reflecting systemic drug exposure and predicting both therapeutic efficacy and adverse reactions. In this study, we successfully developed and validated a rapid, sensitive, and accurate liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analytical method for simultaneous quantification of erythrocyte 6-TG and 6-MMPD concentrations. During method development and validation, we focused on addressing the following five key challenges currently of significant interest in the field: 1) Erythrocytes proved superior to whole blood as biological matrix; 2) Perchloric acid was essential in sample pretreatment; 3) Dual mobile phase additives (HCOOH+NH₄Ac) enhanced chromatographic peak shape and detection sensitivity; 4) Incomplete hydrolysis of 6-TGN and 6-MMPN occurred with heating durations < 60 min; 5) The linear correlation coefficient of the 6-MMPD (6-MMP derivative, 4-amino-5-(methylthio)carbonyl imidazole) standard curve reflected the stable conversion efficiency of 6-MMP. The method exhibited excellent linearity over the concentration ranges of 0.06–50 μmol/L for 6-TG (r = 0.9906) and 0.18–150 μmol/L for 6-MMPD (r = 0.9914). Selectivity, carry-over, intra- and inter-batch accuracy and precision, reproducibility, recovery, matrix effect, and stability all complied with the acceptance criteria outlined in the US Food and Drug Administration (FDA) Bioanalytical Method Validation Guidance (2018). The validated method was successfully employed to quantify 6-TG and 6-MMPD in samples obtained from 20 patients with IBD or ALL.

硫嘌呤是一类广泛用于炎症性肠病（IBD）和急性淋巴细胞白血病（ALL）患者的抗代谢物，它与两种主要的不良反应相关：延迟性骨髓毒性和肝毒性。红细胞活性代谢物6-硫鸟嘌呤核苷酸（6-TGN）和6-甲基巯基嘌呤核苷酸（6-MMPN）可作为反映全身药物暴露的生物标志物，预测治疗疗效和不良反应。在本研究中，我们成功建立并验证了一种快速、灵敏、准确的液相色谱-电喷雾电离-串联质谱（LC-ESI-MS/MS）同时定量红细胞6-TG和6-MMPD浓度的分析方法。在方法开发和验证期间，我们专注于解决以下五个关键挑战，目前该领域的重大兴趣：1)红细胞被证明优于全血作为生物基质；2)样品预处理必须使用高氯酸；3)双流动相添加剂（HCOOH+NH4Ac）增强了色谱峰形和检测灵敏度；4) 6-TGN和6-MMPN在加热时间< 60 min时发生不完全水解；5) 6-MMPD （6-MMP衍生物，4-氨基-5-（甲基硫）羰基咪唑）标准曲线的线性相关系数反映了6-MMP稳定的转化效率。6-TG在0.06 ~ 50 μmol/L （r = 0.9906）和6-MMPD在0.18 ~ 150 μmol/L （r = 0.9914）的浓度范围内具有良好的线性关系。选择性、结转、批内和批间准确度和精密度、重现性、回收率、基质效应和稳定性均符合美国食品和药物管理局（FDA）生物分析方法验证指南（2018）中概述的验收标准。该方法被成功用于定量20例IBD或ALL患者样品中的6-TG和6-MMPD。

{"title":"An optimized LC-ESI-MS/MS assay for erythrocyte 6-TG and 6-MMPD: Addressing critical methodological considerations for thiopurine metabolite monitoring","authors":"Ya-Min Chu , Yuan-Yuan Zhang , Ya-Hui Hu , Jie Wang , Hong-Li Guo , Jin Xu , Yong-Jun Fang , Jing Xu , Xuan-Sheng Ding , Feng Chen","doi":"10.1016/j.jchromb.2025.124872","DOIUrl":"10.1016/j.jchromb.2025.124872","url":null,"abstract":"<div><div>Thiopurines, a class of antimetabolites widely used in patients with inflammatory bowel disease (IBD) and acute lymphoblastic leukemia (ALL), are associated with two predominant adverse effects: delayed myelotoxicity and hepatotoxicity. The erythrocyte-based active metabolites 6-thioguanine nucleotides (6-TGN) and 6-methylmercaptopurine nucleotides (6-MMPN) serve as biomarkers reflecting systemic drug exposure and predicting both therapeutic efficacy and adverse reactions. In this study, we successfully developed and validated a rapid, sensitive, and accurate liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analytical method for simultaneous quantification of erythrocyte 6-TG and 6-MMPD concentrations. During method development and validation, we focused on addressing the following five key challenges currently of significant interest in the field: 1) Erythrocytes proved superior to whole blood as biological matrix; 2) Perchloric acid was essential in sample pretreatment; 3) Dual mobile phase additives (HCOOH+NH<sub>4</sub>Ac) enhanced chromatographic peak shape and detection sensitivity; 4) Incomplete hydrolysis of 6-TGN and 6-MMPN occurred with heating durations < 60 min; 5) The linear correlation coefficient of the 6-MMPD (6-MMP derivative, 4-amino-5-(methylthio)carbonyl imidazole) standard curve reflected the stable conversion efficiency of 6-MMP. The method exhibited excellent linearity over the concentration ranges of 0.06–50 μmol/L for 6-TG (<em>r</em> = 0.9906) and 0.18–150 μmol/L for 6-MMPD (<em>r</em> = 0.9914). Selectivity, carry-over, intra- and inter-batch accuracy and precision, reproducibility, recovery, matrix effect, and stability all complied with the acceptance criteria outlined in the US Food and Drug Administration (FDA) Bioanalytical Method Validation Guidance (2018). The validated method was successfully employed to quantify 6-TG and 6-MMPD in samples obtained from 20 patients with IBD or ALL.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1269 ","pages":"Article 124872"},"PeriodicalIF":2.8,"publicationDate":"2026-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145673397","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}

引用次数: 0

Recent advances in three-dimensional graphene and its derivatives for sample preparation in food contaminant analysis: A review 三维石墨烯及其衍生物在食品污染物分析样品制备中的研究进展

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-01-15 Epub Date: 2025-11-20 DOI: 10.1016/j.jchromb.2025.124862

Xuesheng Zhou , Fangyuan Li , Kun Jiang , Na Wang , Bo Cui

Due to their high specific surface area and extended π-conjugated systems, various three-dimensional graphene and its derivatives have been successfully synthesized and widely utilized for the enrichment of contaminants in food matrices. However, the diversity of synthetic approaches for these materials, coupled with their broad applicability in food contaminant extraction, has limited their effectiveness as a coherent and unified framework for guiding the development of detection methods. Therefore, it is assumed that a systematic review of these synthesis strategies, together with a comprehensive elucidation of the mechanisms underlying the extraction of various contaminants, would provide precise guidance for the design of detection methods based on such advanced materials. Based on this rationale, the review presents a systematic overview of the synthesis methods for three-dimensional graphene and its derivatives, accompanied by a critical evaluation of the advantages and limitations of each approach. Furthermore, it summarizes recent applications of these materials in the enrichment of diverse food contaminants and elucidates the underlying adsorption mechanisms. The results indicate that each synthetic method entails specific limitations: high energy consumption in hydrothermal and solvothermal methods, the use of toxic reagents in chemical reduction method, reliance on cross-linking agents in freeze-drying and three-dimensional printing techniques, and the requirement for sophisticated equipment in chemical vapor deposition. Nevertheless, integrating multiple approaches can effectively overcome the inherent drawbacks of individual methods, as demonstrated by the microwave-assisted hydrothermal method. Moreover, π–π stacking, hydrophobic, electrostatic interactions and hydrogen bonding collectively confer a distinct advantage in adsorption of aromatic food contaminants.

由于其高比表面积和扩展的π共轭体系，各种三维石墨烯及其衍生物已被成功合成，并广泛应用于食品基质中污染物的富集。然而，这些材料的合成方法的多样性，加上它们在食品污染物提取中的广泛适用性，限制了它们作为指导检测方法发展的连贯和统一框架的有效性。因此，假设对这些合成策略的系统回顾，以及对各种污染物提取机制的全面阐明，将为基于这些先进材料的检测方法的设计提供精确的指导。基于这一基本原理，本文对三维石墨烯及其衍生物的合成方法进行了系统概述，并对每种方法的优点和局限性进行了批判性评估。综述了近年来这些材料在各种食品污染物富集中的应用，并阐述了其潜在的吸附机制。结果表明，每种合成方法都有特定的局限性：水热法和溶剂热法的高能耗，化学还原法使用有毒试剂，冷冻干燥和三维打印技术依赖交联剂，化学气相沉积对精密设备的要求。然而，结合多种方法可以有效地克服单个方法的固有缺陷，微波辅助水热法证明了这一点。此外，π -π堆积、疏水、静电相互作用和氢键共同赋予了芳香食品污染物吸附的明显优势。

{"title":"Recent advances in three-dimensional graphene and its derivatives for sample preparation in food contaminant analysis: A review","authors":"Xuesheng Zhou , Fangyuan Li , Kun Jiang , Na Wang , Bo Cui","doi":"10.1016/j.jchromb.2025.124862","DOIUrl":"10.1016/j.jchromb.2025.124862","url":null,"abstract":"<div><div>Due to their high specific surface area and extended π-conjugated systems, various three-dimensional graphene and its derivatives have been successfully synthesized and widely utilized for the enrichment of contaminants in food matrices. However, the diversity of synthetic approaches for these materials, coupled with their broad applicability in food contaminant extraction, has limited their effectiveness as a coherent and unified framework for guiding the development of detection methods. Therefore, it is assumed that a systematic review of these synthesis strategies, together with a comprehensive elucidation of the mechanisms underlying the extraction of various contaminants, would provide precise guidance for the design of detection methods based on such advanced materials. Based on this rationale, the review presents a systematic overview of the synthesis methods for three-dimensional graphene and its derivatives, accompanied by a critical evaluation of the advantages and limitations of each approach. Furthermore, it summarizes recent applications of these materials in the enrichment of diverse food contaminants and elucidates the underlying adsorption mechanisms. The results indicate that each synthetic method entails specific limitations: high energy consumption in hydrothermal and solvothermal methods, the use of toxic reagents in chemical reduction method, reliance on cross-linking agents in freeze-drying and three-dimensional printing techniques, and the requirement for sophisticated equipment in chemical vapor deposition. Nevertheless, integrating multiple approaches can effectively overcome the inherent drawbacks of individual methods, as demonstrated by the microwave-assisted hydrothermal method. Moreover, π–π stacking, hydrophobic, electrostatic interactions and hydrogen bonding collectively confer a distinct advantage in adsorption of aromatic food contaminants.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1269 ","pages":"Article 124862"},"PeriodicalIF":2.8,"publicationDate":"2026-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145578136","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}

引用次数: 0

Method development and validation for quantitative determination of urinary biomarkers of food intake for multiple foods 多种食物摄入尿液生物标志物定量测定方法的开发与验证

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-01-01 Epub Date: 2025-09-15 DOI: 10.1016/j.jchromb.2025.124793

Christoph Hassenberg , Sebastian T. Soukup , Marina Armeni , Achim Bub , Jil V. Cannas , David Fuentes , Otto Savolainen , Stephanie Seifert , Rikard Landberg , Sabine E. Kulling , Carina I. Mack

Biomarkers of food intake (BFIs) have emerged as a promising objective tool to complement traditional self-reported dietary assessment in nutritional research, with the potential to reduce systematic errors and improve accuracy. The development of comprehensive and robust quantification methods for BFIs is essential for widespread application. However, existing methods typically cover only a moderate number of BFIs per method, hindering their wide application in the field. In this study, we present the development and validation of a method for simultaneous quantification of 80 BFIs in urine reflecting 27 foods. The method utilizes a simple sample preparation procedure, followed by separation using both high-performance liquid chromatography (HPLC) on a C18 column and a hydrophilic interaction chromatography (HILIC) column, combined with tandem mass spectrometry in positive and negative mode (HPLC-MS/MS) (individual runs: 6 min). The working range for each analyte was determined in urine samples from a non-randomized, non-blinded nutritional intervention study. The method was validated with respect to selectivity, linearity, robustness, matrix effects, recovery, accuracy, and precision. In total, 44 BFIs could be absolutely quantified without or with only limitations at low concentrations, while 36 BFIs could only be measured semi-quantitatively, including 16 BFIs with limited validation data due to uncertainties. The 80 BFIs represent 27 foods (6 semi-quantitative) frequently consumed in European diets, including 24 plant-derived and 3 animal-derived items. The future implementation of this large-scale BFI quantification method in nutritional studies is expected to demonstrate the benefits of routinely measuring BFIs to improve the accuracy of dietary assessment.

食物摄入生物标志物（bfi）已成为营养研究中补充传统自我报告饮食评估的一种有希望的客观工具，具有减少系统错误和提高准确性的潜力。为了广泛应用，开发全面、可靠的bfi量化方法至关重要。然而，现有的方法通常只覆盖了中等数量的bfi，阻碍了它们在该领域的广泛应用。在这项研究中，我们提出了一种同时定量27种食物尿液中80种bfi的方法的开发和验证。该方法采用简单的样品制备程序，然后使用高效液相色谱（HPLC）（C18柱）和亲水相互作用色谱（HILIC）柱进行分离，并结合串联质谱（HPLC-MS/MS）进行正负模式分离（单个运行时间：6 min）。每种分析物的工作范围是从一项非随机、非盲法营养干预研究的尿液样本中确定的。对方法的选择性、线性、稳健性、矩阵效应、回收率、准确度和精密度进行了验证。总共有44个bfi在低浓度下可以完全定量，没有或只有限制，而36个bfi只能半定量测量，其中16个bfi由于不确定性而验证数据有限。这80个bfi代表了欧洲饮食中经常食用的27种食物（6种半定量），包括24种植物性和3种动物性食物。这种大规模的BFI量化方法在营养研究中的未来实施有望证明常规测量BFI的好处，以提高饮食评估的准确性。

{"title":"Method development and validation for quantitative determination of urinary biomarkers of food intake for multiple foods","authors":"Christoph Hassenberg , Sebastian T. Soukup , Marina Armeni , Achim Bub , Jil V. Cannas , David Fuentes , Otto Savolainen , Stephanie Seifert , Rikard Landberg , Sabine E. Kulling , Carina I. Mack","doi":"10.1016/j.jchromb.2025.124793","DOIUrl":"10.1016/j.jchromb.2025.124793","url":null,"abstract":"<div><div>Biomarkers of food intake (BFIs) have emerged as a promising objective tool to complement traditional self-reported dietary assessment in nutritional research, with the potential to reduce systematic errors and improve accuracy. The development of comprehensive and robust quantification methods for BFIs is essential for widespread application. However, existing methods typically cover only a moderate number of BFIs per method, hindering their wide application in the field. In this study, we present the development and validation of a method for simultaneous quantification of 80 BFIs in urine reflecting 27 foods. The method utilizes a simple sample preparation procedure, followed by separation using both high-performance liquid chromatography (HPLC) on a C18 column and a hydrophilic interaction chromatography (HILIC) column, combined with tandem mass spectrometry in positive and negative mode (HPLC-MS/MS) (individual runs: 6 min). The working range for each analyte was determined in urine samples from a non-randomized, non-blinded nutritional intervention study. The method was validated with respect to selectivity, linearity, robustness, matrix effects, recovery, accuracy, and precision. In total, 44 BFIs could be absolutely quantified without or with only limitations at low concentrations, while 36 BFIs could only be measured semi-quantitatively, including 16 BFIs with limited validation data due to uncertainties. The 80 BFIs represent 27 foods (6 semi-quantitative) frequently consumed in European diets, including 24 plant-derived and 3 animal-derived items. The future implementation of this large-scale BFI quantification method in nutritional studies is expected to demonstrate the benefits of routinely measuring BFIs to improve the accuracy of dietary assessment.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124793"},"PeriodicalIF":2.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145415360","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}

引用次数: 0

A reversed-phase HPLC-based dual-parameter assay for the human urinary kallidinogenase enzyme activity 基于反相高效液相色谱的双参数测定人尿钾碱二酚原酶活性。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-01-01 Epub Date: 2025-11-06 DOI: 10.1016/j.jchromb.2025.124847

Hong-yan Ge , Ya-jun Zhang , Jun-qin Qiao , Li Fan , He-liang Fu , Hong-zhen Lian

In this work, a dual-parameter quantification protocol was successfully developed for determining the enzymatic activity of human urinary kallidinogenase (KN) using reversed-phase high-performance liquid chromatography (RP-HPLC). KN serves as a hydrolytic enzyme to cleave the substrate S-2266 (H-D-Val-Leu-Arg-pNA). Effective separation of the product, substrate, and KN was achieved under optimized chromatographic conditions. By minimizing matrix interference through HPLC separation and using multi-wavelength detection, both selectivity and sensitivity were significantly improved. Moreover, the dual-parameter quantification method, which simultaneously measured the formation of reaction products and the consumption of substrates to reflect KN enzyme activity, markedly improved the reliability of the determination. This method was successfully applied to the activity assay of actual KN samples. The established method has been demonstrated to be effective and reliable with comparable greenness. It provides a powerful tool for the quantitative monitoring of KN activity in biological products and paves the way for its application in complex biological matrices.

本文建立了一种双参数定量方法，用于反相高效液相色谱（RP-HPLC）测定人尿钾碱二酚原酶（KN）的酶活性。KN作为水解酶裂解底物S-2266 （H-D-Val-Leu-Arg-pNA）。在优化的色谱条件下实现了产物、底物和KN的有效分离。通过高效液相色谱分离和多波长检测，减少基质干扰，显著提高了选择性和灵敏度。双参数定量法同时测定反应产物的生成和底物的消耗，反映KN酶的活性，显著提高了测定的可靠性。该方法成功地应用于实际KN样品的活性测定。所建立的方法已被证明是有效和可靠的，具有相当的绿色。为生物制品中KN活性的定量监测提供了有力的工具，为其在复杂生物基质中的应用铺平了道路。

{"title":"A reversed-phase HPLC-based dual-parameter assay for the human urinary kallidinogenase enzyme activity","authors":"Hong-yan Ge , Ya-jun Zhang , Jun-qin Qiao , Li Fan , He-liang Fu , Hong-zhen Lian","doi":"10.1016/j.jchromb.2025.124847","DOIUrl":"10.1016/j.jchromb.2025.124847","url":null,"abstract":"<div><div>In this work, a dual-parameter quantification protocol was successfully developed for determining the enzymatic activity of human urinary kallidinogenase (KN) using reversed-phase high-performance liquid chromatography (RP-HPLC). KN serves as a hydrolytic enzyme to cleave the substrate S-2266 (H-D-Val-Leu-Arg-pNA). Effective separation of the product, substrate, and KN was achieved under optimized chromatographic conditions. By minimizing matrix interference through HPLC separation and using multi-wavelength detection, both selectivity and sensitivity were significantly improved. Moreover, the dual-parameter quantification method, which simultaneously measured the formation of reaction products and the consumption of substrates to reflect KN enzyme activity, markedly improved the reliability of the determination. This method was successfully applied to the activity assay of actual KN samples. The established method has been demonstrated to be effective and reliable with comparable greenness. It provides a powerful tool for the quantitative monitoring of KN activity in biological products and paves the way for its application in complex biological matrices.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124847"},"PeriodicalIF":2.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145524977","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}

引用次数: 0

γ-cyclodextrin based metal organic frameworks as micellar electrokinetic chromatography additive for the chiral separation of dansylated amino acids 以γ-环糊精为基础的金属有机骨架胶束电动色谱添加剂用于丹基化氨基酸的手性分离

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-01-01 Epub Date: 2025-10-28 DOI: 10.1016/j.jchromb.2025.124839

Yan Zhang, Zexi Yu, Xiaotong Zhu, Rongyue Zhang, Nan Li, Min Wang, Juan Qiao

A strategy utilizing oriented γ-cyclodextrin-based metal organic frameworks (γ-CD-MOFs) as chiral additives in micellar electrokinetic chromatography (MEKC) is presented for enhanced enantioseparation of dansylated D,L-amino acids (Dns-D,L-AAs). Its core innovation resides in the fixed orientation of γ-CD within the MOFs structure. This unique characteristic facilitates a shift in chiral recognition from “random interactions” to a “directional and highly efficient recognition” process, thereby substantially enhancing the resolution of chiral separation while overcoming the limitations of free γ-CD, including unstable interactions and poor reproducibility. Systematic optimization of SDS concentration (9.0 mM), γ-CD-MOFs concentration (16.0 mM), and buffer pH (9.5) yielded a maximum resolution (R_s) of 2.2, representing a 50 % improvement over traditional γ-CD. Under these conditions, 14 pairs of Dns-D,L-AAs achieved baseline separation, with 4 additional pairs showing partial separation. Quantitative validation for D,L-methionine (D,L-Met) demonstrated excellent linearity (19.8–1500 μM, r² = 0.999), low limits of detection (6.6 μM) and quantitation (19.8 μM), and high stability (relative standard deviations <5 %). Application to rat plasma samples revealed peak concentration (C_max) of D,L-Met at 90 min post-injection, highlighting its utility in pharmacokinetic studies of amino acid-derived prodrugs (e.g., S-adenosylmethionine). This work introduces oriented γ-CD-MOFs as a transformative chiral additive in MEKC, offering superior selectivity and reproducibility for pharmaceutical and biomedical analyses.

提出了一种利用定向γ-环糊精基金属有机骨架（γ-CD-MOFs）作为胶束电动色谱（MEKC）手性添加剂的策略，以增强丹化D， l -氨基酸（dn -D,L-AAs）的对端分离。其核心创新在于γ-CD在mof结构中的固定取向。这种独特的特性有助于手性识别从“随机相互作用”向“定向和高效识别”过程的转变，从而大大提高了手性分离的分辨率，同时克服了游离γ-CD的局限性，包括相互作用不稳定和再现性差。系统优化SDS浓度（9.0 mM）、γ-CD- mof浓度（16.0 mM）和缓冲液pH(9.5)，最大分辨率（Rs）为2.2，比传统的γ-CD提高了50%。在这些条件下，14对Dns-D， l - aa达到基线分离，另外4对部分分离。定量验证结果表明，D， l -蛋氨酸（D,L-Met）具有良好的线性关系（19.8 ~ 1500 μM, r2 = 0.999），低检出限（6.6 μM）和定量限（19.8 μM），高稳定性（相对标准偏差<； 5%）。应用于大鼠血浆样品显示，注射后90分钟D，L-Met的峰值浓度（Cmax），突出了其在氨基酸衍生的前药（如s -腺苷蛋氨酸）的药代动力学研究中的应用。本研究介绍了定向γ- cd - mof作为MEKC中的一种变革性手性添加剂，为药物和生物医学分析提供了优越的选择性和可重复性。

{"title":"γ-cyclodextrin based metal organic frameworks as micellar electrokinetic chromatography additive for the chiral separation of dansylated amino acids","authors":"Yan Zhang, Zexi Yu, Xiaotong Zhu, Rongyue Zhang, Nan Li, Min Wang, Juan Qiao","doi":"10.1016/j.jchromb.2025.124839","DOIUrl":"10.1016/j.jchromb.2025.124839","url":null,"abstract":"<div><div>A strategy utilizing oriented γ-cyclodextrin-based metal organic frameworks (γ-CD-MOFs) as chiral additives in micellar electrokinetic chromatography (MEKC) is presented for enhanced enantioseparation of dansylated D,L-amino acids (Dns-D,L-AAs). Its core innovation resides in the fixed orientation of γ-CD within the MOFs structure. This unique characteristic facilitates a shift in chiral recognition from “random interactions” to a “directional and highly efficient recognition” process, thereby substantially enhancing the resolution of chiral separation while overcoming the limitations of free γ-CD, including unstable interactions and poor reproducibility. Systematic optimization of SDS concentration (9.0 mM), γ-CD-MOFs concentration (16.0 mM), and buffer pH (9.5) yielded a maximum resolution (<em>R</em><sub>s</sub>) of 2.2, representing a 50 % improvement over traditional γ-CD. Under these conditions, 14 pairs of Dns-D,L-AAs achieved baseline separation, with 4 additional pairs showing partial separation. Quantitative validation for D,L-methionine (D,L-Met) demonstrated excellent linearity (19.8–1500 μM, r<sup>2</sup> = 0.999), low limits of detection (6.6 μM) and quantitation (19.8 μM), and high stability (relative standard deviations <5 %). Application to rat plasma samples revealed peak concentration (C<sub>max</sub>) of D,L-Met at 90 min post-injection, highlighting its utility in pharmacokinetic studies of amino acid-derived prodrugs (e.g., S-adenosylmethionine). This work introduces oriented γ-CD-MOFs as a transformative chiral additive in MEKC, offering superior selectivity and reproducibility for pharmaceutical and biomedical analyses.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124839"},"PeriodicalIF":2.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145415410","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}

引用次数: 0

Characterization and purification of hypoglycemic components from Rubus suavissimus leaf by affinity ultrafiltration, molecular docking, and macroporous resin 采用亲和超滤、分子对接和大孔树脂等方法纯化水荆叶降血糖成分

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-01-01 Epub Date: 2025-10-28 DOI: 10.1016/j.jchromb.2025.124830

Xi Hu , Siye Gao , Hancong Wu , Qingyun He , Jianing Liu , Danyue Zhao , Zhenqiang Wu

This study developed an environmentally friendly method for extracting polyphenols and rubusoside from Rubus suavissimus leaves. Under optimal extraction conditions, the contents of polyphenol and rubusoside reached 91.89 ± 2.39 mg/g and 47.03 ± 1.45 mg/g, respectively. The extract was purified by twelve types of macroporous resins, among which the ADS-7 resin had the highest static adsorption and desorption capacities. Optimized purification parameters for ADS-7 resin were determined by static and dynamic adsorption-desorption experiments and found to be: loading flow rate, 3 BV/h; loading concentration, 10 mg/mL; loading volume, 24 BV; elution solvent, 80 % (v/v) ethanol; elution speed, 3 BV/h; and elution volume, 20 BV. After purification, the content of polyphenols and rubusoside increased by 2.63 and 2.32 times, respectively. Kinetic and thermodynamic analyses revealed distinct adsorption mechanisms: polyphenols followed a pseudo-second-order model, whereas rubusoside followed a pseudo-first-order model; both exhibited spontaneous (ΔG < 0) and endothermic (ΔH > 0) physisorption.

Ultrafiltration-assisted centrifugal fractionation combined with UPLC-ESI-qTOF-MS/MS identified eight potential α-glucosidase inhibitors, including quercetin, apigenin, rutin, ellagic acid, kaempferol-3-O-rutinoside, kaempferol, rubusoside, and steviol. Molecular docking simulations revealed the molecular interactions between these compounds and α-glucosidase.

The enriched extract demonstrated enhanced bioactivity, with a 3.35-fold higher α-glucosidase inhibition, and its IC₅₀ values for DPPH and ABTS⁺ radical scavenging decreased from 142.03 μg/mL to 88.45 μg/mL and from 197.63 μg/mL to 107.63 μg/mL, respectively. These results substantiate the effectiveness of the extraction-purification approach for producing R. suavissimus leaf extracts and suggest their potential application as functional hypoglycemic and antioxidant food ingredients.

本研究建立了一种环境友好的提取水绵叶中多酚和水绵苷的方法。在最佳提取条件下，多酚和冬凌草苷的含量分别为91.89±2.39 mg/g和47.03±1.45 mg/g。采用12种不同的大孔树脂对提取物进行纯化，其中ADS-7树脂的静态吸附和解吸能力最高。通过静态和动态吸附-解吸实验确定了ADS-7树脂的最佳纯化参数：加载流量为3 BV/h；负载浓度，10 mg/mL；装载容积：24bv；洗脱溶剂：80% （v/v）乙醇；洗脱速度：3 BV/h；洗脱体积为20 BV。纯化后，其多酚类和冬冬苷含量分别提高了2.63倍和2.32倍。动力学和热力学分析揭示了不同的吸附机理：多酚类遵循准二级吸附模型，而鲁布索苷类遵循准一级吸附模型；两者均表现自发（ΔG < 0）和吸热（ΔH > 0）物理吸收。超滤辅助离心分离联合UPLC-ESI-qTOF-MS/MS鉴定出8种潜在的α-葡萄糖苷酶抑制剂，包括槲皮素、芹菜素、芦丁、鞣花酸、山奈酚-3- o -芦丁苷、山奈酚、冬冬苷和甜菊醇。分子对接模拟揭示了这些化合物与α-葡萄糖苷酶之间的分子相互作用。强化提取物的生物活性增强，α-葡萄糖苷酶抑制力提高3.35倍，其清除DPPH和ABTS+自由基的IC₅₀值分别从142.03 μg/mL降至88.45 μg/mL和从197.63 μg/mL降至107.63 μg/mL。这些结果证实了提取-纯化法制备山参叶提取物的有效性，并提示其作为功能性降血糖和抗氧化食品原料的潜在应用前景。

{"title":"Characterization and purification of hypoglycemic components from Rubus suavissimus leaf by affinity ultrafiltration, molecular docking, and macroporous resin","authors":"Xi Hu , Siye Gao , Hancong Wu , Qingyun He , Jianing Liu , Danyue Zhao , Zhenqiang Wu","doi":"10.1016/j.jchromb.2025.124830","DOIUrl":"10.1016/j.jchromb.2025.124830","url":null,"abstract":"<div><div>This study developed an environmentally friendly method for extracting polyphenols and rubusoside from <em>Rubus suavissimus</em> leaves. Under optimal extraction conditions, the contents of polyphenol and rubusoside reached 91.89 ± 2.39 mg/g and 47.03 ± 1.45 mg/g, respectively. The extract was purified by twelve types of macroporous resins, among which the ADS-7 resin had the highest static adsorption and desorption capacities. Optimized purification parameters for ADS-7 resin were determined by static and dynamic adsorption-desorption experiments and found to be: loading flow rate, 3 BV/h; loading concentration, 10 mg/mL; loading volume, 24 BV; elution solvent, 80 % (<em>v</em>/v) ethanol; elution speed, 3 BV/h; and elution volume, 20 BV. After purification, the content of polyphenols and rubusoside increased by 2.63 and 2.32 times, respectively. Kinetic and thermodynamic analyses revealed distinct adsorption mechanisms: polyphenols followed a pseudo-second-order model, whereas rubusoside followed a pseudo-first-order model; both exhibited spontaneous (ΔG < 0) and endothermic (ΔH > 0) physisorption.</div><div>Ultrafiltration-assisted centrifugal fractionation combined with UPLC-ESI-qTOF-MS/MS identified eight potential α-glucosidase inhibitors, including quercetin, apigenin, rutin, ellagic acid, kaempferol-3-O-rutinoside, kaempferol, rubusoside, and steviol. Molecular docking simulations revealed the molecular interactions between these compounds and α-glucosidase.</div><div>The enriched extract demonstrated enhanced bioactivity, with a 3.35-fold higher α-glucosidase inhibition, and its IC₅₀ values for DPPH and ABTS<sup>+</sup> radical scavenging decreased from 142.03 μg/mL to 88.45 μg/mL and from 197.63 μg/mL to 107.63 μg/mL, respectively. These results substantiate the effectiveness of the extraction-purification approach for producing <em>R. suavissimus</em> leaf extracts and suggest their potential application as functional hypoglycemic and antioxidant food ingredients.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124830"},"PeriodicalIF":2.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145415411","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}

引用次数: 0

Causal effects of telomere length on ovarian tumor risk: Insights from genetic evidence 端粒长度对卵巢肿瘤风险的因果影响：来自遗传证据的见解

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-01-01 Epub Date: 2025-11-05 DOI: 10.1016/j.jchromb.2025.124845

Tingyu Lang , Xiaolei Liang , Yongxiu Yang

Background

Currently, contradictions remain regarding the causal relationship between telomere length (TL) and ovarian tumors (OT). To elucidate the potential causal relationship, we conducted a two-sample bidirectional Mendelian randomization (MR) study.

Method

All the data were from European populations and obtained from the Genome-Wide Association Study, the FinnGen study, and the Ovarian Cancer Association Consortium. We used inverse variance weighting (IVW) as the primary method, supplemented by other five methods to calculate odds ratios (OR) and 95 % confidence intervals (95 % CI) and checked for pleiotropy. Cochran's Q was used to detect heterogeneity.

Results

The IVW method supported TL as a risk factor for malignant (OR = 1.273, 95 % CI = 1.012–1.602) and benign ovarian tumors (BOTs) (OR = 1.337, 95 % CI = 1.093–1.637). Besides, ovarian cancer subtypes evaluated the strong causal relationship between TL and low malignant potential serous ovarian cancer (LMSOC) by applying the IVW method after Bonferroni correction. There was suggestive evidence for low malignant potential mucinous ovarian cancer, invasive mucinous ovarian cancer, and low-grade serous ovarian cancer. No causal relationships were found for other ovarian cancer subtypes.

Conclusion

Our MR study provides strong genetic evidence that longer telomere length increases risk of specific ovarian tumor subtypes—most robustly for benign ovarian tumors (BOTs) and low malignant potential serous ovarian cancer (LMSOC). For other subtypes, observed associations were suggestive but inconclusive after multiplicity correction, highlighting the etiological heterogeneity of ovarian tumors To optimize treatment regimens and preventive measures for OT, we will continue to thoroughly investigate the potential mechanism of action of these associations in future studies.

目前，关于端粒长度（TL）与卵巢肿瘤（OT）之间的因果关系仍存在矛盾。为了阐明潜在的因果关系，我们进行了一项双样本双向孟德尔随机化（MR）研究。方法所有数据均来自欧洲人群，分别来自全基因组关联研究、FinnGen研究和卵巢癌协会联盟。我们以逆方差加权（IVW）为主要方法，辅以其他5种方法计算比值比（OR）和95%置信区间（95% CI），并检查多效性。Cochran’s Q用于检测异质性。结果IVW方法支持TL是卵巢恶性肿瘤（OR = 1.273, 95% CI = 1.012 ~ 1.602）和良性肿瘤（bot）的危险因素（OR = 1.337, 95% CI = 1.093 ~ 1.637）。此外，通过Bonferroni校正后的IVW方法，卵巢癌亚型评估TL与低恶性潜在浆液性卵巢癌（LMSOC）之间的强因果关系。提示潜在的低恶性粘液性卵巢癌、浸润性粘液性卵巢癌和低级别浆液性卵巢癌。其他卵巢癌亚型没有发现因果关系。结论磁共振研究提供了强有力的遗传学证据，表明端粒长度较长会增加特定卵巢肿瘤亚型的风险，其中良性卵巢肿瘤（bot）和低恶性潜势浆液性卵巢癌（LMSOC）的风险最大。对于其他亚型，观察到的关联是提示性的，但经过多重校正后不确定，突出了卵巢肿瘤的病因异质性。为了优化治疗方案和预防措施，我们将在未来的研究中继续深入研究这些关联的潜在作用机制。

{"title":"Causal effects of telomere length on ovarian tumor risk: Insights from genetic evidence","authors":"Tingyu Lang , Xiaolei Liang , Yongxiu Yang","doi":"10.1016/j.jchromb.2025.124845","DOIUrl":"10.1016/j.jchromb.2025.124845","url":null,"abstract":"<div><h3>Background</h3><div>Currently, contradictions remain regarding the causal relationship between telomere length (TL) and ovarian tumors (OT). To elucidate the potential causal relationship, we conducted a two-sample bidirectional Mendelian randomization (MR) study.</div></div><div><h3>Method</h3><div>All the data were from European populations and obtained from the Genome-Wide Association Study, the FinnGen study, and the Ovarian Cancer Association Consortium. We used inverse variance weighting (IVW) as the primary method, supplemented by other five methods to calculate odds ratios (OR) and 95 % confidence intervals (95 % CI) and checked for pleiotropy. Cochran's Q was used to detect heterogeneity.</div></div><div><h3>Results</h3><div>The IVW method supported TL as a risk factor for malignant (OR = 1.273, 95 % CI = 1.012–1.602) and benign ovarian tumors (BOTs) (OR = 1.337, 95 % CI = 1.093–1.637). Besides, ovarian cancer subtypes evaluated the strong causal relationship between TL and low malignant potential serous ovarian cancer (LMSOC) by applying the IVW method after Bonferroni correction. There was suggestive evidence for low malignant potential mucinous ovarian cancer, invasive mucinous ovarian cancer, and low-grade serous ovarian cancer. No causal relationships were found for other ovarian cancer subtypes.</div></div><div><h3>Conclusion</h3><div>Our MR study provides strong genetic evidence that longer telomere length increases risk of specific ovarian tumor subtypes—most robustly for benign ovarian tumors (BOTs) and low malignant potential serous ovarian cancer (LMSOC). For other subtypes, observed associations were suggestive but inconclusive after multiplicity correction, highlighting the etiological heterogeneity of ovarian tumors To optimize treatment regimens and preventive measures for OT, we will continue to thoroughly investigate the potential mechanism of action of these associations in future studies.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124845"},"PeriodicalIF":2.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145464641","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}

引用次数: 0

Impact of Preprocessing and data analysis strategies on metabolite annotation in biological samples: A mass spectrometry-based metabolomics study using feature-based molecular networking 预处理和数据分析策略对生物样品中代谢物注释的影响：使用基于特征的分子网络进行基于质谱的代谢组学研究

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-01-01 Epub Date: 2025-10-28 DOI: 10.1016/j.jchromb.2025.124828

Qinyu Xiao , Jiamin Yang , Jianjun Xie , Qing Liu , Hongbo Huang , Yiwen Tao , Bo Ding

Metabolomics workflows involve multiple complex steps including sample collection, storage, preparation, metabolite extraction, analytical platforms selection, data acquisition and interpretation. Each step may introduce variability that affects the quality and reliability of metabolomic data. To systematically investigate the effects of these factors on metabolomics outcomes, plasma samples from four different anatomical sites of colon cancer patients were analyzed using Liquid chromatography- Quadrupole-Exactive Orbitrap mass spectrometry (LC-Q-Exactive Orbitrap MS) for untargeted metabolomics. Response surface methodology was employed to optimize the ultrasound-assisted extraction conditions during sample pretreatment. Data analysis strategies were systematically evaluated, including Feature-Based Molecular Networking (FBMN) construction parameters and comparative assessment of different FBMN platforms for metabolite annotation. The optimized extraction conditions were determined as 300 % methanol concentration, sample freezing at −20 °C for 40 min, followed by ultrasonication for 5 min. Sample standardization protocols requiring single-use portioning and limiting freeze-thaw cycles to ≤2–3 cycles were identified as essential for reliable biomarker discovery and therapeutic mechanism exploration. Optimal FBMN construction parameters comprised a 25-min gradient elution time, 50 mm chromatographic column length, and high sample concentration. Comparative evaluation of Global Natural Products Social Molecular Networking (GNPS) and MZmine implementations of FBMN revealed that GNPS was recommended for studies prioritizing comprehensive annotation coverage and discovery-oriented metabolomics, while MZmine was preferred for method development, or applications requiring local processing without external data upload. This study demonstrated that preprocessing and data analysis strategies were critical determinants of data quality in untargeted plasma metabolomics. The findings provided evidence-based recommendations for experimental design, storage conditions, and data handling procedures that can guide protocol standardization and minimize undesired analytical variation in metabolomics studies.

代谢组学工作流程涉及多个复杂步骤，包括样品收集、储存、制备、代谢物提取、分析平台选择、数据采集和解释。每一步都可能引入可变性，影响代谢组学数据的质量和可靠性。为了系统地研究这些因素对代谢组学结果的影响，我们使用液相色谱-四极柱- exactive Orbitrap质谱（LC-Q-Exactive Orbitrap MS）对来自结肠癌患者四个不同解剖部位的血浆样本进行了非靶向代谢组学分析。在样品预处理过程中，采用响应面法优化超声辅助提取条件。系统评估数据分析策略，包括基于特征的分子网络（FBMN）构建参数和不同FBMN平台代谢物注释的比较评估。最佳提取条件为：甲醇浓度为300%，样品在- 20℃冷冻40 min，超声处理5 min。样品标准化方案要求一次性分割，并将冻融周期限制在≤2-3个周期，这对于可靠的生物标志物发现和治疗机制探索至关重要。最佳FBMN构建参数为25 min梯度洗脱时间、50 mm色谱柱长度和高样品浓度。对全球天然产物社会分子网络（Global Natural Products Social Molecular network， GNPS）和MZmine实现FBMN的比较评估表明，GNPS被推荐用于优先考虑综合注释覆盖和以发现为导向的代谢组学的研究，而MZmine更适合于方法开发或需要本地处理而不需要外部数据上传的应用。该研究表明，预处理和数据分析策略是非靶向血浆代谢组学数据质量的关键决定因素。这些发现为实验设计、储存条件和数据处理程序提供了基于证据的建议，可以指导方案标准化，并最大限度地减少代谢组学研究中不希望出现的分析差异。

{"title":"Impact of Preprocessing and data analysis strategies on metabolite annotation in biological samples: A mass spectrometry-based metabolomics study using feature-based molecular networking","authors":"Qinyu Xiao , Jiamin Yang , Jianjun Xie , Qing Liu , Hongbo Huang , Yiwen Tao , Bo Ding","doi":"10.1016/j.jchromb.2025.124828","DOIUrl":"10.1016/j.jchromb.2025.124828","url":null,"abstract":"<div><div>Metabolomics workflows involve multiple complex steps including sample collection, storage, preparation, metabolite extraction, analytical platforms selection, data acquisition and interpretation. Each step may introduce variability that affects the quality and reliability of metabolomic data. To systematically investigate the effects of these factors on metabolomics outcomes, plasma samples from four different anatomical sites of colon cancer patients were analyzed using Liquid chromatography- Quadrupole-Exactive Orbitrap mass spectrometry (LC-Q-Exactive Orbitrap MS) for untargeted metabolomics. Response surface methodology was employed to optimize the ultrasound-assisted extraction conditions during sample pretreatment. Data analysis strategies were systematically evaluated, including Feature-Based Molecular Networking (FBMN) construction parameters and comparative assessment of different FBMN platforms for metabolite annotation. The optimized extraction conditions were determined as 300 % methanol concentration, sample freezing at −20 °C for 40 min, followed by ultrasonication for 5 min. Sample standardization protocols requiring single-use portioning and limiting freeze-thaw cycles to ≤2–3 cycles were identified as essential for reliable biomarker discovery and therapeutic mechanism exploration. Optimal FBMN construction parameters comprised a 25-min gradient elution time, 50 mm chromatographic column length, and high sample concentration. Comparative evaluation of Global Natural Products Social Molecular Networking (GNPS) and MZmine implementations of FBMN revealed that GNPS was recommended for studies prioritizing comprehensive annotation coverage and discovery-oriented metabolomics, while MZmine was preferred for method development, or applications requiring local processing without external data upload. This study demonstrated that preprocessing and data analysis strategies were critical determinants of data quality in untargeted plasma metabolomics. The findings provided evidence-based recommendations for experimental design, storage conditions, and data handling procedures that can guide protocol standardization and minimize undesired analytical variation in metabolomics studies.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124828"},"PeriodicalIF":2.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145415359","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}

引用次数: 0

Bio-inspired ionic liquid design: An advanced and environmentally friendly microextraction method for the selective separation and precise quantification of gallic acid in complex plant-derived matrices 仿生离子液体设计：一种先进且环保的微萃取方法，用于复杂植物源性基质中没食子酸的选择性分离和精确定量。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-01-01 Epub Date: 2025-11-17 DOI: 10.1016/j.jchromb.2025.124859

Mehdi Hosseini , Ebaa Adnan Azooz

Accurate determination of gallic acid (GA), a potent antioxidant present in grapevine leaves, is essential for optimizing extraction processes in pharmaceutical and food applications. Therefore, developing highly selective and sensitive methods for its analysis in complex plant-derived matrices is critical. To address this, a bio-inspired ionic liquid based on the gallic acid structure was designed, specifically the ionic liquid 1-[(3,4,5-trihydroxybenzyl)methyl]pyridinium chloride ([GaPy][Cl]). This tailored ionic liquid enables precise and selective microextraction of gallic acid via a simple and well-defined mechanism, thus enhancing the accuracy of its quantification in complex plant-derived grapevine leaf matrices. The ionic liquid was successfully synthesized and characterized by various techniques, including NMR, FTIR, and elemental analysis. The extraction mechanism of gallic acid from grapevine leaf samples was elucidated through Density Functional Theory (DFT) calculations, which revealed that strong hydrogen bonding at multiple sites is responsible for the highly efficient interactions during the extraction process. Based on systematic experimental evaluations conducted using the developed microextraction method, the proposed approach successfully quantified gallic acid at very low concentrations, achieving a limit of detection (LOD) of 0.13 ng mL⁻¹. The method exhibits excellent precision, with intra-day and inter-day relative standard deviations (RSDs, n = 7) of 1.79 % and 2.74 %, respectively. Furthermore, recovery studies performed on various real samples—including grape leaves, grapes, grapevine stems, and selected aqueous matrices—yielded recoveries ranging from 94.6 % to 99.4 %, confirming the high accuracy and robust analytical performance of the method for the determination of gallic acid in complex matrices.

没食子酸（GA）是葡萄藤叶中一种有效的抗氧化剂，准确测定其含量对优化制药和食品中的提取工艺至关重要。因此，开发高选择性和敏感的方法来分析复杂的植物源性基质是至关重要的。为了解决这个问题，设计了一种基于没食子酸结构的仿生离子液体，即离子液体1-[（3,4,5-三羟基苄基）甲基]氯化吡啶（[GaPy][Cl]）。这种定制的离子液体能够通过简单而明确的机制精确和选择性地微提取没食子酸，从而提高其在复杂植物衍生葡萄藤叶基质中的定量准确性。成功地合成了离子液体，并通过核磁共振、红外光谱和元素分析等多种技术对其进行了表征。利用密度泛函理论（DFT）分析了葡萄叶样品中没食子酸的提取机理，结果表明，在提取过程中，多位点的强氢键是高效相互作用的原因。采用微萃取方法进行了系统的实验评估，该方法在极低浓度下成功地定量了没食子酸，检出限（LOD）为0.13 ng mL-1。方法精密度高，日内、日间相对标准偏差（rsd, n = 7）分别为1.79%和2.74%。此外，对各种实际样品（包括葡萄叶、葡萄、葡萄藤茎和选定的水基质）进行了回收率研究，回收率从94.6%到99.4%不等，证实了该方法在复杂基质中测定没食子酸的高精度和稳健的分析性能。

{"title":"Bio-inspired ionic liquid design: An advanced and environmentally friendly microextraction method for the selective separation and precise quantification of gallic acid in complex plant-derived matrices","authors":"Mehdi Hosseini , Ebaa Adnan Azooz","doi":"10.1016/j.jchromb.2025.124859","DOIUrl":"10.1016/j.jchromb.2025.124859","url":null,"abstract":"<div><div>Accurate determination of gallic acid (GA), a potent antioxidant present in grapevine leaves, is essential for optimizing extraction processes in pharmaceutical and food applications. Therefore, developing highly selective and sensitive methods for its analysis in complex plant-derived matrices is critical. To address this, a bio-inspired ionic liquid based on the gallic acid structure was designed, specifically the ionic liquid 1-[(3,4,5-trihydroxybenzyl)methyl]pyridinium chloride ([GaPy][Cl]). This tailored ionic liquid enables precise and selective microextraction of gallic acid via a simple and well-defined mechanism, thus enhancing the accuracy of its quantification in complex plant-derived grapevine leaf matrices. The ionic liquid was successfully synthesized and characterized by various techniques, including NMR, FTIR, and elemental analysis. The extraction mechanism of gallic acid from grapevine leaf samples was elucidated through Density Functional Theory (DFT) calculations, which revealed that strong hydrogen bonding at multiple sites is responsible for the highly efficient interactions during the extraction process. Based on systematic experimental evaluations conducted using the developed microextraction method, the proposed approach successfully quantified gallic acid at very low concentrations, achieving a limit of detection (LOD) of 0.13 ng mL<sup>−1</sup>. The method exhibits excellent precision, with intra-day and inter-day relative standard deviations (RSDs, <em>n</em> = 7) of 1.79 % and 2.74 %, respectively. Furthermore, recovery studies performed on various real samples—including grape leaves, grapes, grapevine stems, and selected aqueous matrices—yielded recoveries ranging from 94.6 % to 99.4 %, confirming the high accuracy and robust analytical performance of the method for the determination of gallic acid in complex matrices.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124859"},"PeriodicalIF":2.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145558941","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}

引用次数: 0