Journal of Chromatography B最新文献_第4页

Quantitative GC–MS/MS in the clinical analysis of eicosanoids: A critical review and discussion, a 40-years historical retrospect, and possible implications for LC-MS/MS 定量GC-MS /MS在类二十烷酸临床分析中的应用：一个重要的回顾和讨论，40年的历史回顾，以及对LC-MS/MS的可能影响

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-02-01 Epub Date: 2025-12-31 DOI: 10.1016/j.jchromb.2025.124911

Dimitrios Tsikas

Mass spectrometry was a protagonist in the discovery of prostaglandins, thromboxane, leukotrienes and other arachidonic acid-derived molecules, collectively known as the eicosanoids. Mass spectrometry has played a significant role in exploration of their metabolic pathways in humans and animals, in health and disease, and in pharmacotherapy. Clinical researchers in the United States of America and in Europe, in close cooperation with chemist analysts, were the pioneers in the application of gas chromatography–mass spectrometry (GC–MS) and gas chromatography-tandem mass spectrometry (GC–MS/MS) to quantitate eicosanoids and index metabolites in plasma, serum and urine samples from clinical trials by using stable-isotope labeled analogs as internal standards. In the present article, the application of the stable-isotope dilution GC–MS/MS methodology in the quantitative clinical analysis is reviewed. The focus is on prostaglandins, thromboxane, leukotrienes, and their so-called index metabolites for renal and whole-body synthesis of certain eicosanoids such as PGE₂ and its major urinary metabolite (PGE-MUM), respectively. Nowadays, LC-MS/MS, which evolved later than GC–MS/MS, is increasingly used in numerous areas of research, including the eicosanoids in clinical studies. The present work critically discusses the current practice of LC-MS/MS users in the quantitative analysis of eicosanoids in biological samples. While the LC-MS/MS technology offers rapidity and high-throughput analysis, especially due to the renunciation of time-consuming analytical derivatization steps that are required in GC–MS/MS, LC-MS/MS seems to lack sufficient analytical sensitivity, i.e., lower limit of quantitation, for many eicosanoids such as thromboxane B₂ and leukotriene B₄. Reported data on basal concentrations of certain eicosanoids in plasma and urine samples from healthy humans as determined by LC-MS/MS are several orders of magnitude higher than originally reported by pioneering eicosanoid researchers, who developed, validated and used sophisticated, tailored GC–MS- and GC–MS/MS-based analytical methods for individual eicosanoids. Modern eicosanoids researchers would greatly benefit from the milestones and signposts set previously eicosanoids researchers from the very start. A key milestone and signpost is the concentration of primary eicosanoids and their metabolites in plasma, serum and urine samples of healthy humans. Issues for consideration in the GC–MS/MS and LC-MS/MS analysis of eicosanoids are discussed.

质谱法是发现前列腺素、血栓素、白三烯和其他花生四烯酸衍生分子的主要方法，这些分子统称为类二十烷。质谱法在探索人类和动物的代谢途径、健康和疾病以及药物治疗方面发挥了重要作用。美国和欧洲的临床研究人员与化学分析人员密切合作，率先应用气相色谱-质谱法（GC-MS）和气相色谱-串联质谱法（GC-MS /MS），以稳定同位素标记的类似物作为内标，定量临床试验血浆、血清和尿液样品中的类二十烷酸和指数代谢物。本文综述了稳定同位素稀释GC-MS /MS方法在临床定量分析中的应用。重点是前列腺素、血栓素、白三烯及其所谓的指标代谢物，分别用于肾脏和全身合成某些类二十烷，如PGE2及其主要尿液代谢物（PGE-MUM）。目前，LC-MS/MS的发展比GC-MS /MS晚，越来越多地应用于许多研究领域，包括临床研究中的类二十烷类化合物。本工作批判性地讨论了LC-MS/MS用户在生物样品中二十烷类化合物定量分析中的当前实践。虽然LC-MS/MS技术提供了快速和高通量的分析，特别是由于放弃了GC-MS /MS所需的耗时的分析衍生化步骤，LC-MS/MS似乎缺乏足够的分析灵敏度，即定量下限，对于许多类二十烷，如血栓素B2和白三烯B4。通过LC-MS/MS测定的健康人血浆和尿液样本中某些类二十烷的基础浓度的报告数据比最初由开创性的类二十烷研究人员报道的高几个数量级，这些研究人员开发、验证并使用了复杂的、量身定制的基于GC-MS和GC-MS /MS的分析方法来分析单个类二十烷。现代类二十烷酸研究人员将极大地受益于以前的类二十烷酸研究人员从一开始就设定的里程碑和路标。一个关键的里程碑和路标是初级类二十烷酸及其代谢物在健康人血浆、血清和尿液样本中的浓度。讨论了类二十烷类化合物的GC-MS /MS和LC-MS/MS分析中应注意的问题。

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引用次数: 0

Pharmacokinetic assessment and metabolic stability of a novel multi-targeted Clk/Dyrk inhibitor using a validated LC-MS/MS method: In vitro and in vivo insights 基于LC-MS/MS方法的新型多靶点Clk/Dyrk抑制剂的药代动力学评估和代谢稳定性：体外和体内观察

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-02-01 Epub Date: 2025-12-03 DOI: 10.1016/j.jchromb.2025.124888

Maryam Soliman , Noha Mostafa , Sally Tarek Mahmoud , Dalia S. El-Gamil , Mohammad Abdel-Halim , Marwa Fouad

Compound BHBC-01, a promising multi-target inhibitor of Dyrk1A, Dyrk1B, and Clk1, which is based on 5-hydroxybenzo[b]thiophene-2-carboxylic acid benzylamide scaffold, has demonstrated its potential as a novel anticancer agent. To support its further development, this study investigated its pharmacokinetic and stability profiles. An ultra-high performance liquid chromatography-tandem mass spectrometry method was developed and validated for the analysis of compound BHBC-01. Chromatographic separation was achieved using an ACQUITY UPLC BEH C18 column with gradient elution, employing a mobile phase consisting of 0.1 % formic acid in water and acetonitrile at a flow rate of 0.4 mL/min. Diphenhydramine was used as the internal standard. Electrospray ionization in positive ion mode allowed multiple reaction monitoring detection using parent ions ([M + H]⁺) at m/z 314.096 and m/z 256.14, and daughter ions at m/z 120.94 and m/z 167.04 for compound BHBC-01 and diphenhydramine, respectively. The bioanalytical method was validated according to US-FDA guidelines, demonstrating good linearity (50–1000 ng/mL), accuracy, precision, specificity, and stability, with a lower limit of quantification of 50 ng/mL. This method was applied to evaluate the in vitro metabolic stability of compound BHBC-01 in rat liver S9 fraction, revealing a half-life time of 1.95 h and an intrinsic clearance of 11.94 mL/min/kg, indicating stability against biotransformation. Compound BHBC-01 also exhibited significant stability in human plasma, with a half-life time of 57.7 h. Furthermore, gastrointestinal stability studies in simulated gastric and intestinal fluids demonstrated half-lives of 13.75 and 14.11 h, respectively, supporting its suitability for oral administration. In vivo pharmacokinetic studies were conducted in Sprague-Dawley rats following intravenous (5 mg/kg) and oral (30 mg/kg) administration. According to the findings, the compound demonstrates adequate pharmacokinetic characteristics through both intravenous and oral routes. The intravenous route demonstrated a C_max of 545.40 ng/mL, an AUC_0-t of 434.01 ng*h/mL, and a half-life time of 0.66 h. Oral administration showed higher C_max (657 ng/mL), and a half-life time of 7.18 h. The oral bioavailability (F) was calculated to be 19.62 %. Collectively, these findings highlight compound BHBC-01's favorable pharmacokinetic and stability profiles, supporting its potential as a drug candidate for further clinical development as a multi-target anticancer agent.

化合物BHBC-01是一种基于5-羟基苯并[b]噻吩-2-羧酸苄酰胺支架的Dyrk1A、Dyrk1B和Clk1的有前途的多靶点抑制剂，已被证明具有作为新型抗癌药物的潜力。为了支持其进一步开发，本研究考察了其药代动力学和稳定性。建立了化合物BHBC-01的超高效液相色谱-串联质谱分析方法，并进行了验证。色谱分离采用ACQUITY UPLC BEH C18柱，梯度洗脱，流动相为0.1%甲酸水溶液和乙腈，流速为0.4 mL/min。以苯海拉明为内标。在正离子模式下电喷雾电离，对化合物BHBC-01和苯海拉明分别采用M /z 314.096和M /z 256.14的母离子（[M + H]+）和M /z 120.94和M /z 167.04的子离子进行多反应监测检测。根据US-FDA指南对该生物分析方法进行验证，显示出良好的线性（50-1000 ng/mL），准确度，精密度，特异性和稳定性，定量下限为50 ng/mL。采用该方法评价了化合物BHBC-01在大鼠肝脏S9组分中的体外代谢稳定性，发现其半衰期为1.95 h，内在清除率为11.94 mL/min/kg，具有抗生物转化的稳定性。化合物BHBC-01在人血浆中也表现出显著的稳定性，半衰期为57.7 h。此外，在模拟胃液和肠液中的胃肠道稳定性研究表明，半衰期分别为13.75和14.11 h，支持其口服给药的适用性。对Sprague-Dawley大鼠进行了静脉注射（5 mg/kg）和口服（30 mg/kg）的体内药代动力学研究。根据研究结果，该化合物通过静脉注射和口服途径都表现出足够的药代动力学特征。静脉给药Cmax为545.40 ng/mL， AUC0-t为434.01 ng*h/mL，半衰期为0.66 h，口服给药Cmax为657 ng/mL，半衰期为7.18 h，口服生物利用度(F)为19.62%。总的来说，这些发现突出了化合物BHBC-01良好的药代动力学和稳定性，支持其作为多靶点抗癌药物进一步临床开发的候选药物的潜力。

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引用次数: 0

Systematic characterization of in vivo exposure and pharmacokinetic features of Tongmaijiangzhi capsule in humans using integrated HRMS and intelligent data processing 采用综合HRMS和智能数据处理技术系统表征通脉降脂胶囊在人体内暴露和药代动力学特征

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-02-01 Epub Date: 2025-12-22 DOI: 10.1016/j.jchromb.2025.124907

Yuexin Yang , Chunyan Zhu , Qiannan Xu , Bahriman Xarpidin , Dandan Zhang , Hairong Zhang , Xi Luo , Wenjing Tian , HaiFeng Chen , Caisheng Wu

Hyperlipidemia is a major risk factor for cardiovascular diseases and fatty liver. Its incidence has increased in recent years, becoming a major public health concern that poses a serious threat to human health. Tongmaijiangzhi Capsule (TMJZC), a classical pure traditional Chinese medicine (TCM) formulation, has been widely applied in the clinical management of hyperlipidemia with confirmed efficacy and favorable safety. However, its in vivo exposure profile and pharmacokinetic characteristics in humans remain insufficiently characterized, limiting its rational clinical use. In this study, a human exposure-driven analytical strategy was employed, integrating high-resolution mass spectrometry (HRMS), untargeted intelligent data mining, and multi-component quantitative analysis to systematically elucidate the pharmacokinetic behavior and potential bioactive constituents of TMJZC in humans. Using a self-developed untargeted HRMS data-processing platform, a total of 215 TMJZC-related compounds (including 74 prototype constituents and 141 metabolites) were successfully identified in human plasma and urine. Based on human exposure abundance, quality-control components of the herbal medicine, and their circulating forms in vivo, selected seven potential active compounds for further investigation (Cryptochlorogenic acid, Chlorogenic acid, Kaempferol 3-O-β-sophoroside, Hyperoside, Nuciferine, Ginsenoside Rg1, and Notoginsenoside R1). Subsequently, a sensitive and selective multi-component quantification method was subsequently established to systematically characterize their metabolic profiles in humans. Meanwhile, sex-related differences in pharmacokinetics were observed, with the geometric mean ratios (GMRs, female/male) of Nuciferine, Ginsenoside Rg1 and Hyperoside showing substantially higher Cmax (1134.13 %, 447.65 %, 261.53 %,) and AUC_0–∞ (580.47 %, 422.63 %, 290.51 %) in females, respectively. In addition, pharmacological evaluation using hyperlipidemic hepatocyte models (AML12 and HepG2 cells) demonstrated that Hyperoside, Nuciferine, Notoginsenoside R1, and Ginsenoside Rg1 significantly reduced intracellulartriglyceride accumulation. Overall, this study provides the first comprehensive human exposure and pharmacokinetic characterization of TMJZC, highlighting representative circulating constituents and sex-related metabolic differences, and offers a robust analytical basis for quality evaluation, rational clinical application, and further mechanistic studies of this traditional Chinese medicine formulation.

高脂血症是心血管疾病和脂肪肝的主要危险因素。近年来，其发病率有所上升，已成为严重威胁人类健康的重大公共卫生问题。通脉降脂胶囊（TMJZC）是一种经典的纯中药制剂，已广泛应用于临床治疗高脂血症，疗效证实，安全性良好。然而，其体内暴露特征和人体内药代动力学特征仍然不充分，限制了其合理的临床应用。本研究采用人体暴露驱动分析策略，结合高分辨率质谱（HRMS）、非靶向智能数据挖掘和多组分定量分析，系统阐明TMJZC在人体内的药动学行为和潜在的生物活性成分。利用自主开发的非靶向HRMS数据处理平台，在人血浆和尿液中成功鉴定了215种tmjzc相关化合物（包括74种原型成分和141种代谢物）。根据人体暴露丰度、质量控制成分及其在体内循环形式，选择7种潜在活性化合物（隐绿原酸、绿原酸、山奈酚3-O-β-槐苷、金丝桃苷、荷叶碱、人参皂苷Rg1和三七皂苷R1）进行进一步研究。随后，建立了一种灵敏、选择性的多组分定量方法，系统地表征了它们在人体中的代谢谱。同时，观察到药代动力学的性别差异，荷叶碱、人参皂苷Rg1和金丝桃苷的几何平均比（GMRs，女性/男性）在女性中Cmax（1134.13%、447.65%、261.53%）和AUC0 -∞（580.47%、422.63%、290.51%）均显著高于女性。此外，利用高脂血症肝细胞模型（AML12和HepG2细胞）进行的药理学评估表明，金丝桃苷、荷叶碱、三七皂苷R1和人参皂苷Rg1显著降低了细胞内甘油三酯的积累。总体而言，本研究首次提供了TMJZC的全面人体暴露和药代动力学特征，突出了具有代表性的循环成分和性别相关的代谢差异，为该中药制剂的质量评价、合理临床应用和进一步的机制研究提供了有力的分析基础。

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引用次数: 0

Rapid and sensitive detection of Candida albicans using nystatin-functionalized magnetic spheres 制氨抑素功能化磁球快速灵敏检测白色念珠菌。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-02-01 Epub Date: 2025-12-16 DOI: 10.1016/j.jchromb.2025.124896

Zhen-Yu Zhao , Ya-Ning Che , Qian Huang , Han-Wen Yang , Xian-Hua Wang , Lin-Yi Dong

Candida albicans is the leading cause of invasive fungal disease; candidemia carries 30–40 % mortality. It commonly colonizes human mucosa as a commensal, but immunosuppression or disrupted microbiota enables opportunistic infection. Therefore, rapid, reliable diagnostics are vital to improve public health.

In this study, a magnetic solid-phase microextraction (MSPE) technique based on nystatin-functionalized magnetic nanoparticles (Fe₃O₄@NYS) was developed to achieve the dual objectives of efficiently enriching C. albicans rapidly and extracting its deoxyribonucleic acid (DNA). Optimization of capture conditions enabled rapid enrichment of C. albicans with a maximum recovery rate of approximately 90 %. Subsequent to the capture of C. albicans, DNA extraction was performed on the Fe₃O₄@NYS-C. albicans complexes. Notably, Fe₃O₄@NYS facilitated the extraction of a substantial quantity of DNA, with the extraction yield reaching 1200 ng/mg. Therefore, the integration of DNA extraction with quantitative polymerase chain reaction (qPCR) technology enables sensitive and specific detection of C. albicans in real samples. The strategy demonstrated excellent linearity within the concentration range of 4.20 × 10² to 4.20 × 10⁷ CFU mL⁻¹ for C. albicans. The high sensitivity and accuracy of this method were confirmed in honey samples, showcasing a limit of detection (LOD) 420 CFU mL⁻¹, recoveries between 98.1 % and 99.8 %, and relative standard deviations (RSD) not exceeding 2.51 %. This method presents an innovative strategy for point-of-care clinical diagnosis, disease surveillance, and biosafety management, demonstrating substantial potential for the efficient isolation of DNA from complex biological matrices.

白色念珠菌是侵袭性真菌疾病的主要原因；念珠菌的死亡率为30- 40%。它通常以共生的形式定植在人粘膜上，但免疫抑制或微生物群破坏使机会性感染成为可能。因此，快速、可靠的诊断对于改善公共卫生至关重要。本研究建立了一种基于nystatin功能化磁性纳米颗粒（Fe₃O₄@NYS）的磁固相微萃取（MSPE）技术，以实现快速高效富集白色念珠菌和提取其脱氧核糖核酸（DNA）的双重目的。优化捕获条件，使白色念珠菌快速富集，最大回收率约为90%。捕获白色念珠菌后，在Fe₃O₄@NYS-C上进行DNA提取。白色的复合物。值得注意的是，Fe₃O₄@NYS促进了大量DNA的提取，提取率达到1200 ng/mg。因此，将DNA提取与定量聚合酶链反应（qPCR）技术相结合，可以对真实样品中的白色念珠菌进行灵敏、特异的检测。该策略对白色念珠菌在4.20 × 102 ~ 4.20 × 107 CFU mL-1浓度范围内具有良好的线性关系。该方法在蜂蜜样品中具有较高的灵敏度和准确性，检出限（LOD）为420 CFU mL-1，回收率为98.1% ~ 99.8%，相对标准偏差（RSD）不超过2.51%。该方法为即时临床诊断、疾病监测和生物安全管理提供了一种创新策略，展示了从复杂生物基质中有效分离DNA的巨大潜力。

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引用次数: 0

Integrated chemometric and spectrum-effect analysis discriminates Belamcanda chinensis and Iris tectorum in treating A549 lung cancer cells 综合化学计量学和光谱效应分析鉴别了白菖蒲和鸢尾对A549肺癌细胞的治疗作用

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-02-01 Epub Date: 2025-12-01 DOI: 10.1016/j.jchromb.2025.124875

Li-Jun Ruan , Bing-Xiong Yan , Yun-Qiu Wu , Ru-Xin Liu , Shan-Shan Song , Cai-Yun Yao , Feng-Li Huang , Zhi-Jun Song

Belamcanda chinensis (射干, She Gan, SG) and Iris tectorum (川射干, Chuan She Gan, CS), morphologically similar Iridaceae species with overlapping phytochemistry and respiratory applications, are frequently misused in traditional and modern practice. This study established a comprehensive differentiation strategy for SG and CS by integrating chemical characterization with pharmacological evaluation to identify bioactive markers. Twenty-nine herbal samples (11 CS, 18 SG) were analyzed for extraction yields, HPLC fingerprints, isoflavone quantification, multivariate chemometric analysis, in vitro cytotoxicity assessment, and spectrum-effect correlations. CS exhibited significantly higher extraction yields (44.8 % vs. 23.4 %) and total isoflavonoid content (168.7 vs. 42.6 mg/g) than SG, which correlated with stronger cytotoxicity against A549 cells. Chemometric analysis (PCA, PLS-DA and HCA) distinguished species-specific markers: tectoridin/ iristectorin A & B/ iristectorigenin B dominated CS, while irigenin characterized SG. In cytotoxicity assays using A549 cells at concentrations of 50–400 μg/mL, CS extracts showed significantly stronger dose-dependent cytotoxicity (93.6 % to 32.1 % viability reduction) compared to SG extracts (102.8 % to 68.3 %). PLSR-based spectrum-effect relationship analysis at 100/200 μg/mL linked eleven chromatographic peaks to bioactivity, with tectoridin, iristectorin A, and iristectorigenin B demonstrating strong correlations. Experimental validation confirmed these monomers as key cytotoxic agents. In summary, our integrated chemometric and spectrum-effect analysis showed stronger cytotoxicity of CS versus SG in A549 cells, with tectoridin, iristectorin A, and iristectorigenin B identified as potential quality markers. This study establishes a reliable chemometric-bioassay approach for bioactive marker discovery and herb-herb differentiation in traditional medicine. Further validation in diverse models and mechanistic studies is needed.

Belamcanda对(射干,她甘,SG)和虹膜tectorum(川射干,栓她甘,CS),形态相似的鸢尾科物种与重叠的植物化学和呼吸系统应用程序,在传统和现代实践中经常被误用。本研究通过化学表征与药理评价相结合的方法，建立了SG和CS的综合鉴别策略，以鉴定其生物活性标记物。对29份中草药样品（11份CS， 18份SG）进行了提取率、HPLC指纹图谱、异黄酮定量、多元化学计量分析、体外细胞毒性评估和光谱效应相关性分析。CS的提取率（44.8% vs. 23.4%）和总异黄酮含量（168.7 vs. 42.6 mg/g）显著高于SG，且CS对A549细胞具有较强的细胞毒性。化学计量分析（PCA， PLS-DA和HCA）区分了种特异性标记：鸢尾苷/鸢尾苷A和鸢尾苷B/鸢尾苷B以CS为主，而鸢尾苷以SG为特征。在浓度为50 ~ 400 μg/mL的A549细胞的细胞毒性实验中，CS提取物的细胞毒性（活性降低93.6% ~ 32.1%）明显强于SG提取物（活性降低102.8% ~ 68.3%）。在100/200 μg/mL浓度下，基于plsr的光谱效应关系分析将11个色谱峰与生物活性联系起来，鸢尾鸢尾素、鸢尾鸢尾鸢尾素A和鸢尾鸢尾鸢尾素B显示出很强的相关性。实验验证证实这些单体是关键的细胞毒性药物。综上所述，我们的综合化学计量学和光谱效应分析显示，CS比SG在A549细胞中具有更强的细胞毒性，鸢尾鸢尾素、鸢尾鸢尾鸢尾素A和鸢尾鸢尾鸢尾素B被确定为潜在的质量标记。本研究为中药生物活性标志物的发现和中草药鉴别建立了一种可靠的化学计量学-生物测定方法。需要在不同的模型和机制研究中进一步验证。

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引用次数: 0

Simultaneous chiral analysis of DL-asp and DL-α-HB with derivatization in human fingernail by UHPLC-MS/MS: an application in colorectal cancer UHPLC-MS/MS同时手性分析人指甲中衍生DL-asp和DL-α-HB在结直肠癌中的应用

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-02-01 Epub Date: 2025-12-18 DOI: 10.1016/j.jchromb.2025.124895

Yinshuo Xu , Yuanjie Shan , Yuta Nishio , Kenichiro Todoroki , Toshimasa Toyo'oka , Toufeng Jin , Jun Zhe Min

With the development of untargeted metabolomics, studies have shown that alterations in the serum levels of specific amino acids are strongly correlated with the development of early-stage colorectal cancer (CRC). However, the relationship between optically active free amino acids in human nails and the risk of CRC remains unclear. In this study, a highly sensitive UHPLC-MS/MS chiral separation method based on (S)-(−)-DBD-Pro-COCl derivatization was established for the simultaneous detection of DL-Asp, DL-Kyn, DL-α-hydroxybutyric acid (DL-α-HB), and Cystamine in the fingernails of patients with CRC. The derivatization reagents were incubated with DL-amino acids, DL-α-HB and Cyst at 60 °C for 15 min to form diastereomers. Subsequently, an Imtakt cadenza CD-C₁₈ column (3.0 × 150 mm, 3.0 μm) was used for the separation of diastereomers under gradient elution of 10 mM HCOONH₄ in H₂O as mobile phase A and 0.1 % HCOOH in CH₃CN as mobile phase B. The degrees of separation (Rs) ranged from 1.76 to 3.21. R² was >0.9995, indicating good linearity; the limit of detection (LOD) was 25–100 fmol; the interday and intraday precision of DL-Asp and DL-α-HB detection were 0.85 %–4.91 %; and the average recovery ranged from 81.12 % to 110.2 %. However, during the determination process of fingernail samples, only DL-Asp and DL-α-HB were successfully detected, while DL-Kyn and Cyst were not detected. Therefore, this novel method was used to quantify DL-Asp and DL-α-HB in fingernail samples from 32 healthy volunteers (HVs) and 16 patients with CRC (CPs). The results revealed significant differences in D-Asp (p < 0.05), L-Asp (p < 0.01), and L-α-HB (p < 0.05) levels between male HVs and CPs and in D-α-HB (p < 0.05) levels between female HVs and CPs. In addition, the D/L-α-HB ratio (p < 0.01) was significantly different between male HVs and CPs. These findings validate the use of DL-amino acids, DL-α-HB and Cyst from human fingernails as potential diagnostic biomarkers for CRC, providing a basis for the clinical application of human fingernail samples for the noninvasive diagnosis of CRC.

随着非靶向代谢组学的发展，研究表明血清特定氨基酸水平的改变与早期结直肠癌（CRC）的发展密切相关。然而，人类指甲中光学活性游离氨基酸与结直肠癌风险之间的关系尚不清楚。本研究建立了一种基于(S)-(-)- dbd - pro - cocl衍生化的高灵敏度UHPLC-MS/MS手性分离方法，用于同时检测大肠癌患者指甲中DL- asp、DL- kyn、DL-α-羟基丁酸（DL-α- hb）和胱胺。衍生化试剂与DL-氨基酸、DL-α-HB和囊肿在60℃下孵育15min，形成非对映体。采用Imtakt cadenza CD-C18色谱柱（3.0 × 150 mm, 3.0 μm）梯度洗脱，以10 mm HCOONH4水溶液为流动相A， 0.1% HCOOH水溶液为流动相b，分离非对映体，分离度（Rs）为1.76 ~ 3.21。R2为0.9995，线性良好；检出限（LOD） 25 ~ 100 fmol；DL- asp和DL-α-HB检测的日间和日内精密度分别为0.85% ~ 4.91%；平均回收率为81.12% ~ 110.2%。但在指甲样品的检测过程中，仅成功检测到DL- asp和DL-α-HB，未检测到DL- kyn和囊肿。因此，我们采用这种新方法定量了32名健康志愿者（HVs）和16名CRC患者（CPs）指甲样本中的DL- asp和DL-α-HB。结果显示D-Asp (p

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引用次数: 0

Assessment of the benefits of adding differential mobility spectrometry to LC-MS/MS workflows for challenging steroid measurements 在LC-MS/MS工作流程中增加差分迁移度光谱法的好处评估，以用于具有挑战性的类固醇测量。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-02-01 Epub Date: 2025-12-10 DOI: 10.1016/j.jchromb.2025.124893

Kayla Moehnke, Pragya Sharma, Jennifer Kemp, Anthony Maus

Accurate quantification of steroid hormones such as estrogens (estradiol, estrone) and glucocorticoids (cortisol, cortisone) is essential for diagnosing and monitoring endocrine disorders. However, their structural similarity and low physiological concentrations pose analytical challenges in clinical laboratories. This study evaluates the utility of differential mobility spectrometry (DMS) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for improving the specificity and sensitivity of steroid measurements in serum (estrogens) and scalp hair (glucocorticoids) using a SelexION-equipped Sciex 6500+ mass spectrometer coupled to a high throughput Thermo Scientific TLX-2 LC system. DMS significantly reduced chromatographic interferences and enhanced signal-to-noise (S/N) ratios—up to 420 % for estradiol and 210 % for estrone. For cortisol and cortisone in hair, DMS also reduced interferences and increased S/N, as well as improving fragment ion agreement as evidenced by the reduction in fragment ion calculated concentration discrepancies exceeding ±20 % from 18 to 8 samples for cortisol and from 23 to 2 samples for cortisone. These findings support DMS as a complementary technique to LC-MS/MS, offering orthogonal separation and improved analytical performance for steroid hormone quantification in complex biological matrices.

准确定量类固醇激素，如雌激素（雌二醇、雌酮）和糖皮质激素（皮质醇、可的松），对于诊断和监测内分泌紊乱至关重要。然而，它们的结构相似性和低生理浓度给临床实验室的分析带来了挑战。本研究利用selexion - Sciex 6500+质谱联用高通量Thermo Scientific TLX-2 LC系统，评估差分迁移率光谱（DMS）联用液相色谱-串联质谱（LC-MS/MS）在提高血清（雌激素）和头皮（糖皮质激素）中类固醇测量的特异性和敏感性方面的实用性。DMS显著降低了色谱干扰，提高了雌二醇和雌酮的信噪比（S/N），分别高达420%和210%。对于毛发中的皮质醇和可的松，DMS还减少了干扰，增加了S/N，并改善了碎片离子一致性，这一点可以从皮质醇从18个样品减少到8个样品，可的松从23个样品减少到2个样品，碎片离子计算浓度差异超过±20%得到证明。这些发现支持DMS作为LC-MS/MS的补充技术，为复杂生物基质中类固醇激素的定量分析提供正交分离和改进的分析性能。

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引用次数: 0

Chemical composition and bioactivity of Sideritis taurica Stephan ex Wild. (Lamiaceae) leaves: GC/MS analysis, antioxidant and enzyme inhibition activities, and in silico studies 牛黄黄芪的化学成分及生物活性研究。（Lamiaceae）叶片：GC/MS分析，抗氧化和酶抑制活性，以及硅晶硅研究。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-02-01 Epub Date: 2025-12-13 DOI: 10.1016/j.jchromb.2025.124894

Avni Yıldızbaş , Parham Taslimi , Burak Tüzün , Nastaran Sadeghian , Rıfat Kurt , Abdullah İstek

<div><div>Since ancient times, <em>Sideritis taurica</em> and other <em>Sideritis</em> species have been used in traditional medicine in Türkiye and beyond for treating a variety of ailments, including coughs, sore throats, gastrointestinal, respiratory, and urogenital disorders, as well as wounds, colds, and flu, and are believed to possess numerous therapeutic properties such as antispasmodic, analgesic, antibacterial, anti-inflammatory, and antioxidant effects. This study aimed to evaluate the enzyme inhibition and antioxidant activities of various extracts from <em>S. taurica</em> leaves collected from Kurucaşile, Bartın, Türkiye. The extracts were prepared using ethanol, methanol, and hot/cold water extraction methods from leaves that were dried at room temperature and stored in a freezer. Enzyme inhibition activities were assessed against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α-glucosidase, and α-amylase, with IC<sub>50</sub> values calculated. Antioxidant activities were measured using DPPH, ABTS, and CUPRAC assays. Furthermore, ADME/T (Absorption, Distribution, Metabolism, Excretion, and Toxicity) and molecular docking calculations were performed on the phytochemical components of <em>S. taurica</em> to investigate their effects and interactions with human metabolism. These calculations were performed on a number of proteins, including alpha-amylase protein (PDB ID: <span><span>1HNY</span><svg><path></path></svg></span>), AChE protein (PDB ID: <span><span>4M0E</span><svg><path></path></svg></span>), and BChE protein (PDB ID: <span><span>5NN0</span><svg><path></path></svg></span>). The purpose of these calculations was to investigate the interaction between these substances and human metabolism. The results indicated that the ethanol and methanol extracts exhibited the highest inhibition on AChE and BChE (IC<sub>50</sub> values of 73.99 μg/mL and 5.04 μg/mL, respectively). The methanol extracts also demonstrated potent inhibition against α-glucosidase (IC<sub>50</sub> value of 25.81 μg/mL) and α-amylase (IC<sub>50</sub> value of 70.42 μg/mL). Regarding antioxidant activity, the methanol extracts showed the highest radical scavenging activity in the DPPH (87.88 %) and ABTS (99.97 %) tests. Additionally, the methanol extracts stored in the freezer exhibited the best-reducing power in the CUPRAC assay (2.436 ± 0.1669). These findings underscore the potential of <em>S. taurica</em> as a source of natural antioxidants and enzyme inhibitors, suggesting its applicability in the treatment of neurodegenerative diseases such as Alzheimer's disease. In conclusion, extracts obtained from <em>S. taurica</em> leaves, particularly those derived from room temperature-dried leaves, demonstrate significant enzyme inhibition and antioxidant properties. Also, the findings support the consideration of <em>S. taurica</em> as a natural therapeutic source for neurodegenerative diseases and emphasize for further investigation into its active c

自古以来，牛黄黄体炎和其他黄体炎物种就被用于基基ye及其他地区的传统医学中，用于治疗各种疾病，包括咳嗽、喉咙痛、胃肠道、呼吸系统和泌尿生殖系统疾病，以及伤口、感冒和流感，并且被认为具有许多治疗特性，如抗痉挛、镇痛、抗菌、抗炎和抗氧化作用。本研究旨在评价从kuruca、Bartın、trkiye等地采集的金牛花葡萄叶中提取的各种提取物的酶抑制和抗氧化活性。提取液采用乙醇、甲醇和热水/冷水提取的方法从室温下干燥的叶子中提取，并储存在冰箱中。测定对乙酰胆碱酯酶（AChE）、丁基胆碱酯酶（BChE）、α-葡萄糖苷酶和α-淀粉酶的抑制活性，并计算IC50值。采用DPPH、ABTS和CUPRAC测定抗氧化活性。此外，我们还对牛头菌的植物化学成分进行了ADME/T（吸收、分布、代谢、排泄和毒性）和分子对接计算，以研究它们对人体代谢的影响及其相互作用。这些计算是在许多蛋白质上进行的，包括α -淀粉酶蛋白（PDB ID: 1HNY）， AChE蛋白（PDB ID: 4M0E）和BChE蛋白（PDB ID: 5NN0）。这些计算的目的是研究这些物质与人体代谢之间的相互作用。结果表明，乙醇和甲醇提取物对AChE和BChE的抑制作用最强（IC50值分别为73.99 μg/mL和5.04 μg/mL）。甲醇提取物对α-葡萄糖苷酶（IC50值为25.81 μg/mL）和α-淀粉酶（IC50值为70.42 μg/mL）也有较强的抑制作用。在抗氧化活性方面，甲醇提取物在DPPH和ABTS试验中表现出最高的自由基清除活性（87.88%）和（99.97%）。在CUPRAC实验中，冷冻保存的甲醇提取物表现出最好的还原能力（2.436±0.1669）。这些发现强调了牛头葡萄球菌作为天然抗氧化剂和酶抑制剂来源的潜力，表明它在治疗神经退行性疾病如阿尔茨海默病方面的适用性。综上所述，牛头金牛花叶提取物，尤其是室温干燥的牛头金牛花叶提取物，具有显著的酶抑制和抗氧化性能。此外，研究结果支持牛头葡萄球菌作为神经退行性疾病的天然治疗来源的考虑，并强调进一步研究其活性成分和健康益处。

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引用次数: 0

Overcoming the challenge of Protein A resin sanitisation: Evaluating the efficacy of peracetic acid as a novel sanitisation solution for MabSelect SuRe LX resin impacted by spore-forming bacteria 克服蛋白A树脂消毒的挑战：评估过氧乙酸作为一种新型消毒溶液对受孢子形成细菌影响的MabSelect SuRe LX树脂的效果

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-02-01 Epub Date: 2025-12-03 DOI: 10.1016/j.jchromb.2025.124889

Aoife Corr, Aoife Kearney, Aida Aliefendic, Beth Kirschenheiter, Donal Monaghan, Adam Curtin, Pamela O'Brien

Effective sanitisation of Protein A chromatography resin is critical for commercial monoclonal antibody (mAb) production, enabling microbial control and resin reuse. An effective strategy encompasses; resin compatibility with sanitising agents, microbial inactivation, and the removal of residual impurities from the resin. Sodium hydroxide (NaOH) is typically used for resin sanitisation. However, certain spore-forming microorganisms demonstrate resistance to NaOH, even at high concentrations. Alternative sanitising approaches are required to address potential spore-forming bioburden ingress. This paper introduces peracetic acid (PAA) as an alternative to NaOH for remediating MabSelect SuRe LX (MSLX) Protein A resin impacted by spore forming bacteria such as Paenibacillus glucanolyticus, Bacillus subtilis, and Paenibacillus polymyxa. PAA at a 20 mM concentration demonstrated superior microbial control to 0.5 M NaOH. A single 60 min exposure to 20 mM PAA was carried out, followed by continuous cycling until 100 h of NaOH exposure was achieved. Protein A chromatography cycles were conducted at laboratory scale using two distinct mAbs under two loading conditions. A reduction in step yield was observed at the maximum loading condition, suggesting that PAA exposure contributes to a reduction in resin binding capacity. Sufficient binding capacity was maintained when loading at manufacturing-scale relevant ranges for both mAb A & mAb B. Impurity clearance was not impacted following PAA exposure. A manufacturing-scale verification assessment confirmed that a single exposure to 20 mM PAA for 60 min is a suitable remedial treatment for MSLX resin in response to NaOH-resistant bioburden ingress.

蛋白A层析树脂的有效消毒对于商业单克隆抗体（mAb）生产至关重要，可以实现微生物控制和树脂再利用。有效的策略包括：树脂与杀菌剂的相容性，微生物灭活，并从树脂中去除残留杂质。氢氧化钠（NaOH）通常用于树脂消毒。然而，某些孢子形成微生物表现出对NaOH的抗性，即使在高浓度。需要替代的消毒方法来解决潜在的孢子形成的生物负担进入。本文介绍了过氧乙酸（PAA）作为NaOH的替代品，用于修复MabSelect SuRe LX （MSLX）蛋白A树脂，该树脂受到产孢细菌如葡解芽孢杆菌、枯草芽孢杆菌和多粘芽孢杆菌的影响。20 mM浓度的PAA对0.5 M NaOH的微生物控制效果较好。单次暴露于20 mM PAA 60分钟，然后连续循环，直到NaOH暴露100小时。在两种负载条件下，使用两种不同的单抗在实验室规模上进行蛋白A层析循环。在最大负载条件下，观察到阶梯产率的降低，表明PAA暴露有助于树脂结合能力的降低。当在生产规模的相关范围内加载mAb和mAb时，保持足够的结合能力。接触PAA后，杂质清除不受影响。一项生产规模的验证评估证实，单次暴露于20 mM PAA 60分钟是MSLX树脂对耐氢氧化钠生物负荷侵入的一种合适的补救处理。

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引用次数: 0

Online sequential stacking enrichment to verify the relationship between branched chain amino acids in saliva and body mass index by capillary electrophoresis 毛细管电泳在线顺序叠加富集验证唾液支链氨基酸与体重指数的关系

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2026-02-01 Epub Date: 2025-12-15 DOI: 10.1016/j.jchromb.2025.124897

Jianzhu Han , Na Guo , Sijia Zhou, Yijia Chen, Wenqiang Yang, Zhaoyan Wang

The incidence of obesity has increased in recent years, and studies have shown that obese individuals often exhibit elevated branched chain amino acids (BCAAs) in blood. However, the relationship between the salivary BCAA content and obesity remains to be clarified. In this work, BCAAs in saliva were directly determined by capillary electrophoresis with ultraviolet detection (CE-UV) within 15 min, and a sequential stacking enhance combining sweeping and acid barrage stacking (ABS) was employed to enhance sensitivity. Under the optimized conditions, enrichment factors of 49.3, 58.2 and 56.4 were achieved for valine (Val), leucine (Leu), and isoleucine (Ile), respectively. The limits of detection (LODs) for Val, Leu and Ile were 0.28, 0.25 and 0.26 μg/mL, and the limits of quantification (LOQs) were 0.94, 0.83 and 0.88 μg/mL, respectively. Based on salivary BCAA measurements from 16 volunteers, a preliminary positive correlation between salivary BCAA levels and body mass index (BMI) was observed.

近年来，肥胖的发病率有所增加，研究表明，肥胖个体经常表现出血液中支链氨基酸（BCAAs）升高。然而，唾液BCAA含量与肥胖之间的关系尚不清楚。本研究采用毛细管电泳-紫外检测法（CE-UV）在15 min内直接测定唾液中的BCAAs，并采用扫描法和酸弹幕法（ABS）相结合的顺序叠加增强法提高灵敏度。在优化条件下，缬氨酸（Val）、亮氨酸（Leu）和异亮氨酸（Ile）的富集系数分别为49.3、58.2和56.4。Val、Leu和Ile的检出限分别为0.28、0.25和0.26 μg/mL，定量限分别为0.94、0.83和0.88 μg/mL。基于对16名志愿者唾液BCAA的测量，观察到唾液BCAA水平与体重指数（BMI）之间的初步正相关。

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引用次数: 0