Journal of Chromatography B最新文献_第7页

LC-MS-based metabolomics revealed promising role of leukotriene receptor antagonists against colorectal cancer 基于lc - ms的代谢组学揭示了白三烯受体拮抗剂对结直肠癌的良好作用。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-15 DOI: 10.1016/j.jchromb.2025.124824

Farah Hudaib , Sanaa Bardaweel , Wesam Darwish , Salah Abdelrazig , Lina A. Dahabiyeh

Colorectal cancer (CRC) remains one of the leading causes of cancer-related mortality worldwide. Current treatments are often limited by multidrug resistance and significant side effects, underscoring the urgent need for novel therapeutic strategies. Leukotriene receptor antagonists (LTRAs), including montelukast, zafirlukast, and pranlukast, have shown promising anticancer potential; however, their underlying molecular mechanisms remain insufficiently understood. In this study, the antiproliferative effects of montelukast, zafirlukast, and pranlukast on CRC cell lines (HCT-116 and Caco-2) were evaluated using cell viability and proliferation assays. Montelukast and zafirlukast were further examined using a liquid chromatography-mass spectrometry (LC-MS)-based metabolomics to assess any possible metabolic alterations in HCT-116 cells, along with flow cytometry to determine their effects on cell death. Montelukast and zafirlukast were found to significantly inhibit the cell proliferation of HCT-116 in a dose-dependent manner, with IC₅₀ values of 57.4 μM and 73.28 μM, respectively. Flow cytometry demonstrated that apoptosis was the predominant mode of cell death with both drugs. Neither montelukast nor zafirlukast showed any significant in vitro antioxidant effects. LC-MS Metabolomics revealed that montelukast and zafirlukast induced distinct metabolic changes with 47 and 34 significantly altered metabolites, respectively. Montelukast notably affected mannosamine, 5-oxo-L-proline, acetyl-phenylalanine, acetoin, and taurine, implicating pathways related to amino acid metabolism, redox homeostasis, and mitochondrial function. In contrast, zafirlukast impacted nucleotide-related metabolites, including inosine, adenine, cytidine, deoxyguanosine, and itaconate, highlighting nucleotide biosynthesis and immunometabolism disruptions. These findings demonstrate the antiproliferative potential of montelukast and zafirlukast in CRC and provide new insights into their distinct molecular mechanisms of action. This supports their potential repurposing as adjunctive therapies in CRC treatment.

结直肠癌（CRC）仍然是全球癌症相关死亡的主要原因之一。目前的治疗往往受到多药耐药和显著副作用的限制，强调迫切需要新的治疗策略。白三烯受体拮抗剂（LTRAs），包括孟鲁司特、扎非鲁司特和普鲁司特，已经显示出有希望的抗癌潜力；然而，其潜在的分子机制仍未得到充分的了解。在这项研究中，孟鲁司特、扎非鲁司特和普鲁司特对CRC细胞系（HCT-116和Caco-2）的抗增殖作用通过细胞活力和增殖试验进行了评估。使用基于液相色谱-质谱（LC-MS）的代谢组学进一步检测孟鲁司特和扎非鲁司特，以评估HCT-116细胞中任何可能的代谢改变，并使用流式细胞术确定它们对细胞死亡的影响。发现孟鲁司特和扎非鲁司特以剂量依赖的方式显着抑制HCT-116的细胞增殖，IC₅0值分别为57.4 μM和73.28 μM。流式细胞术显示凋亡是两种药物的主要细胞死亡方式。孟鲁司特和扎非鲁司特均未显示出明显的体外抗氧化作用。LC-MS代谢组学显示孟鲁司特和扎非鲁司特诱导了明显的代谢变化，分别有47种和34种代谢物显著改变。孟鲁司特显著影响甘露胺、5-氧- l-脯氨酸、乙酰-苯丙氨酸、乙酰蛋白和牛磺酸，涉及氨基酸代谢、氧化还原稳态和线粒体功能相关的途径。相反，zafirlukast影响核苷酸相关代谢物，包括肌苷、腺嘌呤、胞苷、脱氧鸟苷和衣康酸，突出核苷酸生物合成和免疫代谢中断。这些发现证明了孟鲁司特和扎非鲁司特在结直肠癌中的抗增殖潜力，并为其独特的分子作用机制提供了新的见解。这支持了它们作为CRC治疗辅助疗法的潜力。

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引用次数: 0

Facile construction of hydrophilic core-shell structured magnetic covalent organic framework for efficient magnetic solid-phase extraction of five glucocorticoids from milk sample 亲水性核壳结构磁性共价有机骨架的快速构建，用于高效磁固相萃取牛奶样品中的五种糖皮质激素。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-12 DOI: 10.1016/j.jchromb.2025.124822

Zhi-Peng Li , Chen-Ri Su , Long-Xin Yang , Han-Yue Li , Han Zhang , Ji-Fei Yang , Xian-Hua Wang , Yun-Chao Liu , Rui-Xue Ran

Glucocorticoids play a key role in a variety of physiological processes, but their extensive use in the environment has brought potential health hazards. Herein, it is of great necessity to develop a rapid and efficient method for the detection of glucocorticoids. In this work, a hydrophilic core-shell structured magnetic covalent organic framework (HMCOF) was fabricated via a post-modification strategy for the efficient magnetic solid-phase extraction (MSPE) of five glucocorticoids from tap water and milk samples. The HMCOF featured a Fe₃O₄@SiO₂ magnetic core encapsulated by a porous COF shell modified with polyethylene glycol, endowing it with a hydrophilic outer layer, porous structure and sufficient paramagnetism. The adsorption studies showed that HMCOF exhibited high adsorption capacities (50.77–80.25 mg g⁻¹) for glucocorticoids. Notably, HMCOF retained 80 % of its adsorption capacity after 5 cycles, confirming its reusability. Under the optimal conditions of MSPE, the HMCOF-based MSPE-UPLC method was developed to test five glucocorticoids, which demonstrated good linearity (5–150 ng mL⁻¹, R² ≥ 0.9991), low detection limits (0.2–1.4 ng mL⁻¹) and satisfactory spiked recovery rates (89.5–114.2 %) with intra-day variability below 4.3 % and inter-day precision within 6.3 %. The method underscores the potential of HMCOF serving as an adsorbent for MSPE, providing a promising approach for the analysis of glucocorticoids within food products.

糖皮质激素在多种生理过程中发挥着关键作用，但其在环境中的广泛使用带来了潜在的健康危害。因此，开发一种快速有效的糖皮质激素检测方法是十分必要的。在这项工作中，通过后修饰策略制备了亲水性核壳结构磁性共价有机骨架（HMCOF），用于从自来水和牛奶样品中高效地提取五种糖皮质激素。HMCOF的特点是Fe₃O₄@SiO₂磁芯被聚乙二醇修饰的多孔COF壳包裹，使其具有亲水性外层、多孔结构和足够的顺磁性。吸附研究表明，HMCOF对糖皮质激素具有较高的吸附量（50.77 ~ 80.25 mg g-1）。值得注意的是，HMCOF在5次循环后仍保持了80%的吸附容量，证实了其可重复使用。在最佳MSPE条件下，建立了基于hmcof的MSPE- uplc检测5种糖皮质激素的方法，线性良好（5 ~ 150 ng mL-1, R2≥0.9991），检出限低（0.2 ~ 1.4 ng mL-1），加标回收率（89.5 ~ 114.2%），日内变异性小于4.3%，日内精密度在6.3%以内。该方法强调了HMCOF作为MSPE吸附剂的潜力，为食品中糖皮质激素的分析提供了一种有前途的方法。

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引用次数: 0

Hypoglycemic potential of American ginseng saponins: inhibition of α-amylase and improvement of insulin resistance 西洋参皂苷的降糖潜能：抑制α-淀粉酶和改善胰岛素抵抗。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-10 DOI: 10.1016/j.jchromb.2025.124813

Liwen Liang , Yunlong Guo , Xu Yao , Guangzhi Cai , Xiaokang Liu , Jiyu Gong

Diabetes Mellitus (DM) represents a significant global health challenge, necessitating the development of novel therapeutic agents with reduced adverse effects for effective management. This study investigated the hypoglycemic potential of total saponins derived from American ginseng (TSAG), focusing on its inhibitory activity against α-amylase and its modulatory effects on insulin resistance in HepG2 cells. Results from the α-amylase inhibition assay demonstrated that TSAG elicited significant, concentration-dependent inhibition. Enzyme kinetic analysis revealed that TSAG functions as a reversible, mixed-type inhibitor of α-amylase. In insulin-resistant HepG2 cells, TSAG significantly enhanced glucose consumption and promoted glycogen synthesis, indicating its capacity to ameliorate insulin sensitivity. Ultra-high-performance liquid chromatography coupled with quadrupole orbitrap mass spectrometry (UHPLC-Q-Orbitrap/MS) analysis identified 40 ginsenoside constituents within TSAG. Molecular docking studies demonstrated high-affinity binding interactions between these key ginsenosides and α-amylase, corroborating their contribution to the observed inhibitory effects. Collectively, these findings highlight the therapeutic potential of American ginseng saponins in diabetes management, particularly in improving postprandial glycemic control and enhancing insulin sensitivity.

糖尿病（DM）是一个重大的全球健康挑战，需要开发新的治疗药物，减少不良反应的有效管理。本研究探讨了西洋参总皂苷（TSAG）的降糖潜能，重点研究了其对α-淀粉酶的抑制作用及其对HepG2细胞胰岛素抵抗的调节作用。α-淀粉酶抑制实验结果表明，TSAG具有显著的浓度依赖性抑制作用。酶动力学分析表明，TSAG是一种可逆的混合型α-淀粉酶抑制剂。在胰岛素抵抗的HepG2细胞中，TSAG显著增加葡萄糖消耗，促进糖原合成，表明其改善胰岛素敏感性的能力。超高效液相色谱-四极杆轨道阱质谱联用（UHPLC-Q-Orbitrap/MS）分析鉴定出人参皂苷的40种成分。分子对接研究表明，这些关键人参皂苷与α-淀粉酶之间存在高亲和力的结合作用，证实了它们对观察到的抑制作用的贡献。总的来说，这些发现强调了西洋参皂苷在糖尿病治疗中的治疗潜力，特别是在改善餐后血糖控制和提高胰岛素敏感性方面。

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引用次数: 0

Secretory expression and optimization of type II L-asparaginase from Pseudomonas sp. PCH199 假单胞菌PCH199型l -天冬酰胺酶的分泌表达及优化

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-08 DOI: 10.1016/j.jchromb.2025.124814

Subhash Kumar , Virender Kumar , Dharam Singh

L-asparaginase (L-ASNase) is used to treat acute lymphoblastic leukemia and acrylamide reduction in food. Presently, recombinant L-ASNase formulations are produced in the cytoplasmic fractions of E. coli. Here, we describe an efficient approach for secretory expression of recombinant L-ASNase II (Pg-ASNase II) with pelB signal peptide in the E. coli expression host. Pg-asn II was successfully cloned and expressed in pET-26b(+) having pelB leader sequence for periplasmic expression. It led to the successful secretion of Pg-ASNase II in the supernatant. However, a significant amount of Pg-ASNase II was also left in the cytoplasmic space. The protein secretion increased from 0.33 to 0.77 mg mL^-1 with 0.2 % Tween 80 in the medium. Pg-ASNase II was purified in a single chromatographic step using a Q-sepharose column with 8.0 mg L⁻¹ purified protein production and 60.0 U mg^-1 Pg-ASNase II activity. It demonstrated a 75 % yield and a 15-fold increase in purification. The current study reported an increased extracellular L-ASNase expression compared to the wild organism and helped reduce the cost of the downstream process. Hence, this research can contribute to upcoming investigations of periplasmic Pg-ASNase II for therapeutic applications.

l -天冬酰胺酶（L-ASNase）用于治疗急性淋巴细胞白血病和减少食物中的丙烯酰胺。目前，重组L-ASNase制剂是在大肠杆菌的细胞质部分中生产的。在此，我们描述了一种在大肠杆菌表达宿主中分泌表达含有pelB信号肽的重组L-ASNase II （Pg-ASNase II）的有效方法。成功克隆出Pg-asn II，并在pET-26b（+）中表达，具有pelB领导序列，可在质周表达。这导致Pg-ASNase II在上清液中成功分泌。然而，大量的Pg-ASNase II也留在细胞质间隙中。当培养基中添加0.2%的Tween 80时，蛋白质分泌量从0.33 mg mL-1增加到0.77 mg mL-1。Pg-ASNase II采用Q-sepharose柱纯化，纯化蛋白产量为8.0 mg L-1，活性为60.0 U mg- 1pg - asnase II。它证明了75%的产量和15倍的净化。目前的研究报告了与野生生物相比，细胞外L-ASNase表达增加，有助于降低下游过程的成本。因此，本研究可以为即将开展的质周Pg-ASNase II的治疗应用研究做出贡献。

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引用次数: 0

Innovative multifunctional tag system for protein purification and analytical characterization 用于蛋白质纯化和分析表征的创新多功能标签系统。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-08 DOI: 10.1016/j.jchromb.2025.124811

María Jesús Leopold, Verónica Ferrando , Ricardo Kratje, Marcos Oggero, Natalia Ceaglio

Epitope tag–monoclonal antibody (mAb) systems are essential tools in biotechnology. We developed a novel system based on the mGMOP peptide—derived from GM-CSF with six O-glycosylation sites and an N-terminal APARSPS epitope—and the in-house mAb CC1H7, which binds this epitope, in single or multiple copies, with enhanced affinity under high ionic strength. The mGMOP–mAb CC1H7 system was applied to immunoaffinity chromatography, Western blot, and competitive ELISA. Optimization used two engineered interferon variants: one with a single tag (mGMOP–IFN) and another with four tags (mGMOP3–IFN–mGMOP). A third variant, mGMOP3–EPO–mGMOP, was constructed using erythropoietin for further validation. Immunoaffinity purification was optimized via factorial design, testing different salts. The final protocol used 1 M Na₂SO₄ for loading and 50 mM sodium phosphate at pH 11 for elution, achieving 80–92 % recovery and 100 % purity. Competitive ELISA was optimized using a Box-Behnken design, yielding low LOD and LOQ. Western blotting confirmed detection of both IFN variants, also with low LOD and LOQ. All methods were successfully applied to tagged EPO. The platform enables standardized workflows for characterizing recombinant proteins without requiring protein-specific antibodies, and offers a versatile approach for protein bioprocessing.

表位标签-单克隆抗体（mAb）系统是生物技术中必不可少的工具。我们开发了一种基于mGMOP肽（源自GM-CSF，具有6个o糖基化位点和一个n端APARSPS表位）和内部单克隆抗体CC1H7的新系统，该单克隆或多拷贝结合该表位，在高离子强度下具有增强的亲和力。mgop - mab CC1H7系统应用于免疫亲和层析、Western blot和竞争性ELISA。优化使用了两种工程干扰素变体：一种带有单个标签（mgmopp - ifn），另一种带有四个标签（mGMOP3-IFN-mGMOP）。第三个变体mGMOP3-EPO-mGMOP使用促红细胞生成素构建以进一步验证。通过析因设计优化免疫亲和纯化，检测不同盐类。最终方案采用1 M硫酸钠（Na₂SO₄）和50 mM磷酸钠（pH 11）洗脱，回收率为80- 92%，纯度为100%。竞争性ELISA采用Box-Behnken设计优化，LOD和LOQ均较低。Western blotting证实检测到两种IFN变体，同样具有低LOD和LOQ。所有方法均成功应用于标记EPO。该平台实现了标准化的工作流程，无需蛋白质特异性抗体即可表征重组蛋白，并为蛋白质生物处理提供了一种通用的方法。

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引用次数: 0

Accurate determination of fluoxetine and its metabolites in Mediterranean mussel organs based on solid phase extraction clean-up and HPLC-HRMS 固相萃取净化- HPLC-HRMS法精确测定地中海贻贝器官中氟西汀及其代谢物

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-08 DOI: 10.1016/j.jchromb.2025.124808

E. Lemaire , E. Gomez , D. Rosain , F. Courant

Fluoxetine, a widely prescribed antidepressant, is frequently detected in marine environments, exposing marine invertebrates to this emerging contaminant. Due to its potential to bioaccumulate and undergo biotransformation, it is essential to quantify both the parent compound and its metabolites for accurate risk assessment. Furthermore, tissue-specific measurements are crucial for identifying organ-specific accumulation that may cause localized toxic effects. An analytical method was then developed to quantify fluoxetine and three of its metabolites in individual mussel tissues (soft tissues, gills, digestive glands) using a minimal sample size (40 or 400 mg). Four solid-phase extraction sorbents were evaluated: MCX, HLB, C18, and Phree phospholipid removal. The Phree sorbent was selected as the optimal cleanup method. Further optimization involved testing two extraction solvents (acetonitrile and methanol) and two extraction techniques: ultrasonic-assisted extraction and bead-based extraction. Identification and quantification were performed using HPLC-Orbitrap-MS, relying on accurate mass measurements of selected MS/MS fragments. The method demonstrated satisfactory Method Quantification Limits (MQL) for fluoxetine (1.2, 10.5, and 10.4 μg/kg dw) and its main metabolite, norfluoxetine (5.1, 25.6, and 24.3 μg/kg dw) in soft tissues, digestive glands, and gills, respectively. These MQL values align with literature data, especially given the low sample size and the expression of concentrations on a dry weight basis. The final method allows the sensitive quantification of fluoxetine and its metabolites from small volumes, facilitating the assessment of their distribution across three target organs. These advancements support the development of toxicokinetic/toxicodynamic (TK/TD) models, contributing to environmental risk assessment of emerging contaminants.

氟西汀是一种广泛使用的抗抑郁药，经常在海洋环境中检测到，使海洋无脊椎动物暴露于这种新出现的污染物中。由于其具有生物积累和生物转化的潜力，因此对母体化合物及其代谢物进行量化以进行准确的风险评估至关重要。此外，组织特异性测量对于识别可能导致局部毒性作用的器官特异性积累至关重要。然后开发了一种分析方法，使用最小样本量（40或400毫克）对单个贻贝组织（软组织、鳃、消化腺）中的氟西汀及其三种代谢物进行定量。评估了四种固相萃取吸附剂：MCX， HLB， C18和Phree磷脂去除。选择Phree吸附剂作为最佳的清除方法。进一步的优化包括测试两种提取溶剂（乙腈和甲醇）和两种提取技术：超声波辅助提取和珠状提取。使用HPLC-Orbitrap-MS进行鉴定和定量，依赖于所选MS/MS片段的精确质量测量。该方法在软组织、消化腺和鳃中分别检测到氟西汀（1.2、10.5和10.4 μg/kg dw）及其主要代谢物去甲氟西汀（5.1、25.6和24.3 μg/kg dw）的定量限（MQL）满意。这些MQL值与文献数据一致，特别是考虑到低样本量和以干重为基础的浓度表达。最后一种方法允许小体积的氟西汀及其代谢物的敏感定量，便于评估它们在三个目标器官中的分布。这些进展支持毒物动力学/毒物动力学（TK/TD）模型的发展，有助于对新出现的污染物进行环境风险评估。

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引用次数: 0

A fast GC–MS/MS method for the simultaneous measurement of key metabolites of peroxisomal beta-oxidation and ether lipid biosynthesis in human fibroblasts 同时测定人成纤维细胞过氧化物酶体β -氧化和醚类脂质生物合成关键代谢物的快速GC-MS /MS方法

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-06 DOI: 10.1016/j.jchromb.2025.124806

Karin Preindl , Thomas Stimpfl , Gunda Koellensperger , Fabian Dorninger , Johannes Berger , Christoph Wiesinger

Peroxisomes are subcellular compartments that host a variety of metabolic pathways, including the chain shortening of fatty acids (FAs) by beta-oxidation and certain steps in the formation of ether lipids. Here, we describe the development of a GC–MS/MS-based method for the simultaneous and reproducible determination of key metabolites of these pathways, also including less common FA species related to peroxisomal metabolism that are typically not part of standard analytical methods.

We for the first time utilize 1-chlorobutane for the extraction of FAs as an effective alternative to commonly used extraction solvents. 1-Chlorobutane offers a broader polarity range than hexane and lower toxicity relative to chloroform with solvent consumption of less than one mL per sample.

Six saturated long to very long-chain FAs, nine polyunsaturated FAs (PUFAs), two dicarboxylic FAs and 1-O-octadecyl glycerol (ODG, batyl alcohol) were extracted simultaneously. The method was validated using fibroblasts and for the majority of FA species accuracies ranged from 80 to 110 % with precision values (CV %) from 6 to 20 %. The measurement of ODG is for the first time described as marker for the estimation of the cellular ether lipid synthesis rate. The suitability of the method was demonstrated by the analysis of primary human fibroblasts from controls and individuals with peroxisomal disorders. This cell type represents a widely used model system for the investigation of peroxisomal metabolism and disease, thus rendering our protocol a valuable addition to the toolkit for studying peroxisomal pathways.

过氧化物酶体是承载多种代谢途径的亚细胞室，包括通过β -氧化缩短脂肪酸（FAs）链和形成醚类脂质的某些步骤。在这里，我们描述了一种基于GC-MS / ms的方法的发展，用于同时和可重复性地测定这些途径的关键代谢物，也包括与过氧化物酶体代谢相关的不太常见的FA物种，这些物种通常不是标准分析方法的一部分。我们首次利用1-氯丁烷作为一种有效的萃取溶剂来萃取脂肪酸。1-氯丁烷的极性范围比己烷宽，毒性比氯仿低，每个样品的溶剂消耗量小于1 mL。同时提取了6个饱和长链至甚长链脂肪酸，9个多不饱和脂肪酸（PUFAs）， 2个二羧基脂肪酸和1- o -十八烷基甘油（ODG, batyl alcohol）。用成纤维细胞验证了该方法，对大多数FA物种的准确度在80%到110%之间，精度值（CV %）在6%到20%之间。ODG的测定首次被描述为估计细胞醚脂质合成速率的标记物。通过分析来自对照组和过氧化物酶体疾病患者的原代人成纤维细胞，证明了该方法的适用性。这种细胞类型代表了研究过氧化物酶体代谢和疾病的广泛使用的模型系统，因此使我们的方案成为研究过氧化物酶体途径的工具包的有价值的补充。

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引用次数: 0

Recommendation of a milder cleaning-in-place method for Cytiva's second-generation protein L resin MabSelect VL 推荐Cytiva第二代蛋白L树脂MabSelect VL的温和就地清洗方法。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-04 DOI: 10.1016/j.jchromb.2025.124810

Ju Qu, Yifeng Li, Yan Wan

Protein L-based media are a type of affinity resin whose application in antibody purification is only secondary to Protein A resins. As Protein L resins exhibit stronger byproduct separation capabilities than Protein A resins, they are particularly preferred when purifying complex molecules such as bispecific antibodies (bsAbs). Despite the growing use of Protein L resins, comprehensive evaluations of their lifetime and cleaning-in-place (CIP) methods are generally lacking. In the current study, we examined MabSelect VL, Cytiva's second-generation Protein L resin, for its alkaline stability. In specific, the resin's performance was evaluated after 80 cycles of sanitization with 0.1 M sodium hydroxide (NaOH). In addition, the binding behavior of monomer and aggregates to aged resin was studied to understand the change in resin ligands during use. According to the data, 0.1 M NaOH is not well tolerated by MabSelect VL, and its repeated treatment causes performance degradation. Therefore, we recommend a milder sanitization method, 0.05 M NaOH, which can be combined with 1 % benzyl alcohol to improve microbial control.

蛋白l基介质是一种亲和树脂，其在抗体纯化中的应用仅次于蛋白a树脂。由于蛋白L树脂比蛋白A树脂具有更强的副产物分离能力，因此它们在纯化复杂分子（如双特异性抗体（bsab））时特别受欢迎。尽管蛋白L树脂的使用越来越多，但通常缺乏对其使用寿命和就地清洗（CIP）方法的全面评估。在目前的研究中，我们检测了MabSelect VL， Cytiva的第二代蛋白L树脂，其碱性稳定性。以0.1 M氢氧化钠（NaOH）消毒80次后，对树脂的性能进行了评价。此外，研究了单体和聚集体与老化树脂的结合行为，以了解树脂配体在使用过程中的变化。数据显示，MabSelect VL对0.1 M NaOH的耐受性不佳，反复处理会导致性能下降。因此，我们推荐一种较温和的消毒方法，0.05 M NaOH，可与1%苄醇结合使用，以改善微生物控制。

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引用次数: 0

Retention time prediction of forensic compounds using ensemble machine learning and molecular descriptors 基于集成机器学习和分子描述符的法医化合物保留时间预测。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-02 DOI: 10.1016/j.jchromb.2025.124812

Asena Avci Akca Ph.D. , Sefa Akca Ph.D.

Retention time (RT) prediction can greatly improve the efficiency of chromatographic workflows in forensic toxicology, especially in high-throughput or non-targeted analytical workflows. In the present study, we compare the performance of four ensemble machine learning models—Random Forest (RF), Extra Trees, XGBoost, and LightGBM—in predicting RTs of 229 structurally diverse forensic compounds. Each compound was represented by a minimal set of RDKit-derived descriptors and an extended feature space that combines Mordred descriptors and Morgan circular fingerprints. All RTs were experimentally measured under standardized reversed-phase liquid chromatographic conditions. Model performance was evaluated using coefficient of determination (R²) and root-mean-square error (RMSE). Results show that models trained on extended descriptors (>2000 molecular features) outperformed those trained on basic descriptors, with XGBoost showing the highest predictive power (R² = 0.718, RMSE = 1.23). Feature importance analysis showed that RTs are not only affected by global molecular properties like hydrophobicity and size but also by topological and electronic features. These results highlight the value of ensemble learning in RT prediction and demonstrate its practical utility in compound screening and chromatographic method development in forensic toxicology.

保留时间（RT）预测可以大大提高法医毒理学色谱工作流程的效率，特别是在高通量或非靶向分析工作流程中。在本研究中，我们比较了四种集成机器学习模型——随机森林（random Forest， RF）、Extra Trees、XGBoost和lightgbm——在预测229种结构不同的法医化合物的RTs中的性能。每个化合物都由rdkit派生的描述符的最小集合和一个扩展的特征空间来表示，该特征空间结合了莫德雷德描述符和摩根圆形指纹。所有rt均在标准化反相液相色谱条件下进行实验测量。采用决定系数（R2）和均方根误差（RMSE）评价模型的性能。结果表明，基于扩展描述符（bbb2000分子特征）训练的模型优于基于基本描述符训练的模型，其中XGBoost的预测能力最高（R2 = 0.718, RMSE = 1.23）。特征重要性分析表明，RTs不仅受疏水性和大小等整体分子性质的影响，还受拓扑和电子特征的影响。这些结果突出了集成学习在RT预测中的价值，并展示了其在法医毒理学中化合物筛选和色谱方法开发中的实际应用。

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引用次数: 0

Screening biomarkers for diagnosis of COPD by multiplex-high-resolution mass spectrometry based pseudotargeted lipidomics 基于多分辨率质谱的假靶向脂质组学筛选COPD诊断的生物标志物

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-02 DOI: 10.1016/j.jchromb.2025.124809

Yunfan Zhao , Yang Xie , Hulei Zhao , Jinyan Wu , Xuemei Xu , Liuyue Yin , Yanmin Shi , Jianya Yang , Peng Zhao , Qingzhou Guan , Yan Du , Suyun Li , Jiansheng Li , Xinguang Liu

Triple quadrupole (QQQ) MS based pseudotargeted lipidomics combines high coverage and quantitative accuracy, is commonly established by selecting the most responsive lipid ion pairs identified from high-resolution mass spectrometry (HRMS), then sending them to QQQ MS for quantification by multiple reaction monitoring. Due to the low resolution of QQQ MS, it may result in faulty peak identification in integration and method transition from HRMS to QQQ MS, thus, directly establish pseudotargeted lipidomics on HRMS is needed to improve the accuracy of present methods. We propose a method for rapidly screening high-resolution ion pairs for constructing multiplex-HRMS-based pseudotargeted lipidomics (MHPL). Firstly, high-resolution precursor and product ions for lipid quantification were obtained by HRMS. To solve the problem of lower acquisition speed in HRMS, we used the multiplex mode to simultaneously fragment co-eluting lipid precursor ions within one isolation window, scan the MS/MS mass spectra, and quantify the unique product ions (UPI) of co-eluting lipid (defined as Quan-PIs). Meanwhile, we provide a group of scripts for searching for lipid Quan-PIs in multiplex mode. The proposed MHPL strategy could ensure accurate quantification of 460 lipids within 5 injections, and was applied in serum differential lipid discovery for chronic obstructive pulmonary disease (COPD) patients. As a result, 47 differential lipids were found to comprise a potential biomarker panel for COPD diagnosis. The lipid identification and quantification in this work were conducted in the same instrument, and faulty peak identification was considerably reduced in integration and when transitioning methods from HRMS to QQQ MS.

基于三重四极杆（QQQ）质谱的伪靶向脂质组学具有高覆盖率和定量准确性，通常通过选择高分辨率质谱（HRMS）鉴定的最敏感的脂质离子对，然后将其发送到QQQ质谱进行多反应监测定量来建立。由于QQQ质谱的分辨率较低，在整合过程中可能出现峰识别错误，方法从HRMS过渡到QQQ质谱，因此需要在HRMS上直接建立伪靶向脂质组学，以提高现有方法的准确性。我们提出了一种快速筛选高分辨率离子对的方法，用于构建基于多重hrms的假靶向脂质组学（MHPL）。首先，利用HRMS获得了高分辨率的脂质定量前体和产物离子；为了解决HRMS采集速度较慢的问题，我们采用多重模式在一个分离窗口内同时对共洗脱脂质前体离子进行片段化，扫描MS/MS质谱，并定量共洗脱脂质的唯一产物离子（UPI）（定义为quan - pi）。同时，我们提供了一组多路搜索脂质全pi的脚本。所提出的MHPL策略可确保在5次注射内准确定量460种脂质，并应用于慢性阻塞性肺疾病（COPD）患者的血清脂质鉴别发现。结果，发现47种不同的脂质组成了COPD诊断的潜在生物标志物面板。本工作中的脂质鉴定和定量在同一台仪器上进行，在整合和从HRMS过渡到QQQ MS时，大大减少了错误峰鉴定。

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