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From biosensing to herbal discovery: A nanozyme cascade platform for acetylcholinesterase monitoring and inhibitor screening in synthetic and natural sources 从生物传感到草药发现:一个用于乙酰胆碱酯酶监测和抑制剂筛选的纳米酶级联平台
IF 6.1 1区 化学 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2026-07-01 Epub Date: 2026-02-09 DOI: 10.1016/j.talanta.2026.129523
Shuang Cai , Bao-Wen Fan , Jia Qin , Cheng-Yang Pu , Ya-Jian Qin , Cong-Ting Wu , Wen Xu , Jing Li
Early diagnosis and therapeutic intervention for Alzheimer's disease (AD) necessitate advanced tools for detecting acetylcholinesterase (AChE) activity and screening AChE inhibitors (AChEIs). This study developed a novel bimetallic MOF nanozyme, Cu–NH2–88B(Fe), exhibiting significant peroxidase-like activity. This nanozyme was integrated with AChE to construct a dual-enzyme cascade biosensing platform, which achieved highly sensitive AChE detection, with a detection limit of 0.01 mU/mL, and demonstrated excellent accuracy in fetal bovine serum (spiked recoveries: 92.07–111.09%). Additionally, the platform also enabled quantitative assessment of synthetic AChEIs, determining IC50 values for donepezil (9.85 nM), neostigmine (1.41 μM), huperzine A (6.21 μM), and galantamine (611.31 μM), all of which exhibited broad linear ranges and high sensitivity. All four compounds exhibited a broad linear range and excellent sensitivity. Innovatively, the platform was applied to screen AChE inhibitory activity in seven traditional Chinese medicines (TCMs). Crucially, when comparing four extraction methods, ultrasound-assisted extraction (UAE) proved most effective in liberating active compounds. The UAE-obtained extract of Poria cocos (Yunnan) showed the strongest inhibition, achieving an AChE inhibition rate of 39.03% at 0.01 mg/mL. Notably, the screening results across different species, origins, and extraction methods (including the superior UAE) showed high consistency with the classical Ellman method, validating the platform's reliability. This study not only provides a low-cost, easy-to-operate, sensitive, and reliable analytical strategy for AChE activity detection and AChE inhibitor development, but more importantly, it successfully applies nanozyme technology to the complex system of TCMs, offering strong technical support for the screening and preliminary identification of potential anti-AD active components from TCM resources.
阿尔茨海默病(AD)的早期诊断和治疗干预需要先进的工具来检测乙酰胆碱酯酶(AChE)活性和筛选AChE抑制剂(AChEIs)。本研究开发了一种新型双金属MOF纳米酶Cu-NH2-88B (Fe),具有明显的过氧化物酶样活性。将该纳米酶与乙酰胆碱酯酶结合构建双酶级联生物传感平台,实现了乙酰胆碱酯酶的高灵敏度检测,检出限为0.01 mU/mL,在胎牛血清中具有良好的准确度(加标回收率为92.07 ~ 111.09%)。此外,该平台还可以定量评价合成的AChEIs,测定了多奈哌齐(9.85 nM)、新斯的明(1.41 μM)、石杉碱A (6.21 μM)和加兰他明(611.31 μM)的IC50值,均具有宽线性范围和高灵敏度。四种化合物均具有较宽的线性范围和优良的灵敏度。创新地将该平台应用于7种中药中AChE抑制活性的筛选。关键是,在比较四种提取方法时,超声辅助提取(UAE)被证明在释放活性化合物方面最有效。在0.01 mg/mL浓度下,阿联酋提取的茯苓提取物对乙酰胆碱酯酶的抑制作用最强,抑制率为39.03%。值得注意的是,不同物种、来源和提取方法(包括优质UAE)的筛选结果与经典的Ellman方法高度一致,验证了平台的可靠性。本研究不仅为AChE活性检测和AChE抑制剂开发提供了一种低成本、易操作、灵敏、可靠的分析策略,更重要的是将纳米酶技术成功应用于中药复杂体系,为中药资源中潜在抗ad活性成分的筛选和初步鉴定提供了强有力的技术支持。
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引用次数: 0
Rapid magnetic solid-phase extraction approach employing a modified graphene oxide nanomaterial for the determination of 20 pharmaceuticals and transformation products in treated urban wastewater 采用改性氧化石墨烯纳米材料的快速磁固相萃取法测定处理过的城市废水中的20种药物和转化产物
IF 6.1 1区 化学 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2026-07-01 Epub Date: 2026-02-07 DOI: 10.1016/j.talanta.2026.129521
Ana Belén Martínez-Piernas , Irene Sánchez-Trujillo , Carlos Vereda-Alonso , María del Mar López-Guerrero , Elisa Isabel Vereda-Alonso
A magnetic solid-phase extraction (MSPE) method based on a green magnetic graphene oxide material (d-M@GO) was developed and validated for the simultaneous determination of 20 contaminants of emerging concern (CECs), including pharmaceuticals and transformation products, in wastewater using liquid chromatography–high-resolution mass spectrometry (LC-HRMS). The adsorption behaviour of analytes on d-M@GO was investigated through kinetic and equilibrium modelling, indicating mass-transfer-limited adsorption consistent with a linear isotherm. A two-zone kinetic model, accounting for fast and slow adsorption regions, provided the best fitting for most compounds, confirming the coexistence of easily accessible and diffusion-limited adsorption sites.
The MSPE procedure was optimised to establish the most efficient adsorption and elution conditions. Method validation in treated urban wastewater demonstrated its suitability as a green alternative to conventional solid-phase extraction. Eighteen of the twenty analytes showed recoveries between 70% and 120%, with RSDs below 20%. Low method quantification limits (MQLs) were determined, ranging from 2 to 20 ng/L. Application of the method for the analysis of five wastewater samples enabled the quantification of up to 19 analytes at concentrations ranging from 6.37 to 1321 ng/L. In addition, non-target screening using MS-DIAL open-source software (a platform for untargeted metabolomics and lipidomics data processing) expanded the analytical scope, allowing the tentative identification of 24 additional CECs, five of which were confirmed using reference standards. The combination of target and non-target analyses demonstrates the capability of the method for comprehensive monitoring of CECs in wastewater. With a 60–70% lower carbon footprint than conventional SPE approaches, the d-M@GO-MSPE–LC-HRMS workflow represents a robust, rapid, and sustainable solution.
基于绿色磁性氧化石墨烯材料(d-M@GO)的磁固相萃取(MSPE)方法被开发并验证,用于使用液相色谱-高分辨率质谱(LC-HRMS)同时测定废水中的20种新关注污染物(CECs),包括药品和转化产品。通过动力学和平衡模型研究了分析物在d-M@GO上的吸附行为,表明传质限制吸附符合线性等温线。考虑快速和慢速吸附区域的两区动力学模型对大多数化合物提供了最佳拟合,证实了容易接近和扩散限制的吸附位点共存。优化了MSPE的吸附和洗脱条件。方法在处理后的城市废水中验证了其作为传统固相萃取的绿色替代品的适用性。其中18种加样回收率在70% ~ 120%之间,rsd < 20%。测定了低定量限(MQLs),范围为2 ~ 20 ng/L。应用该方法对5个废水样品进行分析,可以定量分析多达19种浓度范围为6.37至1321 ng/L的分析物。此外,使用MS-DIAL开源软件(非靶向代谢组学和脂质组学数据处理平台)进行非靶标筛选扩大了分析范围,允许初步鉴定24个额外的CECs,其中5个使用参考标准得到确认。目标分析和非目标分析相结合,证明了该方法综合监测废水中CECs的能力。与传统的SPE方法相比,d-M@GO-MSPE -LC-HRMS的碳足迹降低了60-70%,是一种强大、快速、可持续的解决方案。
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引用次数: 0
Immunosensors for dual tumor biomarker detection based on ternary electrochemiluminescence using confined CeO2 nanozyme as Co-reactant enhancers for luminol-O2 system 基于受限CeO2纳米酶作为鲁米诺- o2体系助反应增强剂的三元电化学发光双肿瘤生物标志物检测免疫传感器。
IF 6.1 1区 化学 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2026-07-01 Epub Date: 2026-02-08 DOI: 10.1016/j.talanta.2026.129524
Chao Lu , Xue Fan , Fengna Xi , Yucheng Zhou
Sensitive and parallel individual detection of multiple tumor biomarkers is critical for early diagnosis and therapeutic monitoring of pancreatic cancer. Herein, a ternary electrochemiluminescence (ECL) system was developed by employing nanochannel-confined cerium dioxide (CeO2) nanozyme as co-reactant enhancers for the luminol-dissolved oxygen (DO) system, enabling parallel individual immunoassays of carcinoembryonic antigen (CEA) and carbohydrate antigen 199 (CA199). Vertically-ordered mesoporous silica film (VMSF) was rapidly grown on an indium tin oxide (ITO) electrode, forming nanochannel array with thickness of 97 nm and diameters of 2.7 nm. CeO2 nanozyme was synthesized in situ within the VMSF nanochannels via electrochemical deposition. Transmission electron microscopy (TEM), high-angle annular dark-field scanning TEM (HAADF-STEM), and energy-dispersive X-ray spectroscopy (EDX) elemental mapping confirmed CeO2 nanozyme was confined in nanochannels with high dispersion. CeO2 nanozyme exhibited peroxidase-like catalytic activity and enhanced the ECL intensity of the luminol-DO system by 13.9-fold under near-neutral conditions through catalysis of both oxygen reduction and luminol oxidation. After derivatization of the VMSF outer surface with epoxy groups, antibodies specific to CEA or CA199 were covalently immobilized to construct two immunorecognition interfaces. Target binding formed antigen-antibody complexes, increasing interfacial resistance and hindering diffusion of the ECL emitters, enabling a signal-off immunoassay mode. The resulting immunosensors displayed wide detection linear ranges from 0.1 pg mL−1 to 100 ng mL−1 for CEA and 0.1 × 10−3 mU mL−1 to 1.0 × 104 mU mL−1 for CA199, with limit of detection (LOD) of 1.3 fg mL−1 and 0.5 × 10−4 mU mL−1, respectively. Both immunosensors exhibited good selectivity, reproducibility, and storage stability.
多种肿瘤生物标志物的敏感和平行个体检测对于胰腺癌的早期诊断和治疗监测至关重要。本文采用纳米通道限制的二氧化铈(CeO2)纳米酶作为鲁米诺-溶解氧(DO)体系的助反应增强剂,建立了三元电化学发光(ECL)体系,实现了癌胚抗原(CEA)和碳水化合物抗原199 (CA199)的平行个体免疫测定。垂直有序介孔硅膜(VMSF)在氧化铟锡(ITO)电极上快速生长,形成了厚度为97 nm、直径为2.7 nm的纳米通道阵列。采用电化学沉积的方法在VMSF纳米通道内原位合成了CeO2纳米酶。透射电子显微镜(TEM)、高角环形暗场扫描电镜(HAADF-STEM)和能量色散x射线能谱(EDX)元素图谱证实了CeO2纳米酶被限制在高色散的纳米通道中。CeO2纳米酶表现出类似过氧化物酶的催化活性,在近中性条件下通过催化氧还原和鲁米诺氧化,使鲁米诺- do体系的ECL强度提高了13.9倍。在VMSF外表面环氧基衍生化后,将CEA或CA199特异性抗体共价固定,构建两个免疫识别界面。靶标结合形成抗原-抗体复合物,增加界面阻力并阻碍ECL发射器的扩散,从而实现信号关闭免疫分析模式。所得免疫传感器对CEA的检测线性范围为0.1 pg mL-1 ~ 100 ng mL-1,对CA199的检测线性范围为0.1 × 10-3 mU mL-1 ~ 1.0 × 104 mU mL-1,检测限(LOD)分别为1.3 fg mL-1和0.5 × 10-4 mU mL-1。两种免疫传感器均具有良好的选择性、重复性和储存稳定性。
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引用次数: 0
Hollow Sn-doped ZnO nanotubes for synergistically enhanced sensing performance toward 3-hydroxy-2-butanone with preliminary exploration of a multi-parameter intelligent system 空心掺锡ZnO纳米管协同增强对3-羟基-2-丁酮的传感性能与多参数智能系统的初步探索。
IF 6.1 1区 化学 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2026-07-01 Epub Date: 2026-02-05 DOI: 10.1016/j.talanta.2026.129512
Xiao-Xue Lian, Jing-Wen Liu, Yan Li, Shu-Mei Yang
Targeted detection of the 3-hydroxy-2-butanone (3H-2B) biomarker represents a feasible and reliable strategy for monitoring Listeria monocytogenes, a highly virulent foodborne pathogen that poses severe threats to public health and food safety. The significance of this study lies in addressing the urgent demand for rapid, sensitive, and on-site detection of Listeria monocytogenes, as traditional detection methods are often time-consuming, labor-intensive, and unsuitable for field applications. Herein, we successfully prepared Sn-doped ZnO hollow nanotubes via an electrospinning process, and further designed a multi-parameter intelligent sensing system based on this material for portable 3H-2B detection. A key improvement of our work is the synergistic integration of Sn doping and morphological engineering, which dramatically enhances gas-sensing performance: Sn doping introduces a high density of oxygen vacancies and free charge carriers, while the tailored hollow nanotube configuration maximizes the accessibility of active sites and facilitates rapid gas diffusion across the sensing interface. The optimized 3% Sn-doped ZnO sensor exhibits excellent 3H-2B sensing performance, with an optimal response of 155.1 to 100 ppm 3H-2B at 200 °C, a rapid response time of 1 s, and a low detection limit of 100 ppb, significantly outperforming undoped ZnO and other Sn-doped counterparts. This sensing system shows promising application prospects in food safety supervision, catering industry, and food processing workshops, enabling rapid on-site monitoring of Listeria monocytogenes contamination. This study not only establishes Sn-doped ZnO hollow nanotubes as a superior sensing platform for 3H-2B but also opens a new avenue for the rapid, sensitive detection of foodborne pathogens, providing technical support for food safety guarantee.
3-羟基-2-丁酮(3H-2B)生物标志物的靶向检测是监测单核增生李斯特菌的一种可行和可靠的策略,李斯特菌是一种严重威胁公共卫生和食品安全的高毒力食源性病原体。本研究的意义在于解决了快速、灵敏、现场检测单核增生李斯特菌的迫切需求,传统的检测方法往往耗时、费力,不适合现场应用。本文通过静电纺丝工艺成功制备了掺锡ZnO空心纳米管,并在此基础上设计了多参数智能传感系统,用于便携式3H-2B检测。我们工作的一个关键改进是锡掺杂和形态工程的协同集成,这极大地提高了气敏性能:锡掺杂引入了高密度的氧空位和自由载流子,而定制的空心纳米管配置最大化了活性位点的可达性,并促进了气体在传感界面上的快速扩散。优化后的3% sn掺杂ZnO传感器具有优异的3H-2B传感性能,在200°C下的最佳响应为155.1 ~ 100 ppm 3H-2B,快速响应时间为1 s,检测限低至100 ppb,显著优于未掺杂ZnO和其他sn掺杂ZnO传感器。该传感系统在食品安全监管、餐饮行业、食品加工车间等领域具有广阔的应用前景,可实现单核增生李斯特菌污染的快速现场监测。本研究不仅建立了锡掺杂ZnO空心纳米管作为3H-2B的优越传感平台,而且为食源性致病菌的快速、灵敏检测开辟了新的途径,为食品安全保障提供了技术支持。
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引用次数: 0
Potential adoption of electrochemical biosensors for cancer DNA biomarker detection in liquid biopsies: A systematic review 电化学生物传感器在液体活检中用于癌症DNA生物标志物检测的潜力:系统综述。
IF 6.1 1区 化学 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2026-07-01 Epub Date: 2026-02-03 DOI: 10.1016/j.talanta.2026.129488
Anouk Peymen , Thijs Van der Snickt , Alejandro Valverde , Scott Ailliet , Karen Zwaenepoel , Pieter Mestdagh , Karolien De Wael

Background

The detection of actionable DNA biomarkers in liquid biopsies is essential for advancing precision oncology. While conventional techniques such as PCR and NGS are highly accurate, their high cost, complexity, and slow turnaround time limit widespread clinical applications. Electrochemical biosensors present a promising alternative through their portability, affordability, and rapid analysis capabilities.

Objective

This systematic review evaluates the current status and potential clinical adoption of electrochemical biosensors for detecting cancer DNA biomarkers in liquid biopsies, focusing on their sensitivity and specificity compared to standard PCR-based methods.

Methods

A comprehensive search across PubMed, Scopus, and Web of Science was conducted up to August 2025, yielding 1723 articles. After applying PRISMA guidelines with strict inclusion and exclusion criteria, 31 studies were selected. The quality of the selected studies was assessed using the SURE checklist. Data on biosensor design, amplification strategies, detection limits, and clinical applicability were extracted.

Results

Most biosensors achieved femtomolar to attomolar detection limits (LODs), reaching the required sensitivity limits for germline and somatic mutations respectively These LODs were achieved primarily via signal amplification strategies using redox mediators, nanomaterials, and strand displacement assays. Target amplification was less common and did not demonstrate consistent superiority. Specificity was enhanced through modified probes, enzymatic cleavage, magnetic separation, and blocking agents, though assessment methods varied. In general, covalently immobilized probes improved stability and sensitivity. However, most assays were validated in buffer with synthetic DNA, limiting clinical relevance.

Conclusions

With respect to their performance, electrochemical biosensors exhibit considerable potential for clinical integration. In particular when integrating different signal amplification strategies, such as a synergistic approach of redox mediators and nanocomposites, or combining target and signal amplification to reach the required sensitivity limits. Nonetheless, their clinical integration requires evaluation in complex biological samples, more thorough selectivity studies, and rigorous validation on patient samples. When standard validation assays can be provided and performed, electrochemical biosensing assays hold great promise for the detection of cancer DNA biomarkers in liquid biopsies.
背景:液体活检中可操作的DNA生物标志物的检测对于推进精确肿瘤学至关重要。虽然PCR和NGS等传统技术非常准确,但它们的高成本、复杂性和缓慢的周转时间限制了广泛的临床应用。电化学生物传感器通过其便携性、可负担性和快速分析能力提供了一个有前途的替代方案。目的:本系统综述了电化学生物传感器在液体活检中检测癌症DNA生物标志物的现状和潜在的临床应用,重点介绍了电化学生物传感器与基于pcr的标准方法相比的敏感性和特异性。方法:对PubMed、Scopus和Web of Science进行综合检索,截至2025年8月,共检索到1723篇文章。在应用PRISMA指南和严格的纳入和排除标准后,选择了31项研究。所选研究的质量使用SURE检查表进行评估。提取了生物传感器设计、扩增策略、检测限和临床适用性的数据。结果:大多数生物传感器达到了飞摩尔到原子摩尔的检测限(lod),分别达到了生殖系和体细胞突变所需的灵敏度限。这些lod主要是通过使用氧化还原介质、纳米材料和链位移测定的信号放大策略实现的。靶扩增是不常见的,并没有表现出一贯的优势。特异性通过改良探针、酶裂解、磁分离和阻断剂增强,尽管评估方法各不相同。总的来说,共价固定探针提高了稳定性和灵敏度。然而,大多数分析都是在合成DNA的缓冲液中验证的,限制了临床相关性。结论:就其性能而言,电化学生物传感器具有相当大的临床应用潜力。特别是当整合不同的信号放大策略时,例如氧化还原介质和纳米复合材料的协同方法,或者将目标和信号放大结合起来以达到所需的灵敏度限制。然而,它们的临床整合需要在复杂的生物样本中进行评估,进行更彻底的选择性研究,并对患者样本进行严格的验证。当可以提供和执行标准验证分析时,电化学生物传感分析在液体活检中检测癌症DNA生物标志物方面具有很大的前景。
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引用次数: 0
Magnetic bead-assisted extraction combined with ID-LC-MS/MS for simultaneous quantification of fat-soluble vitamins A, D, E, and K in human serum 磁珠辅助提取联合ID-LC-MS/MS同时定量测定人血清中脂溶性维生素A、D、E和K
IF 6.1 1区 化学 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2026-07-01 Epub Date: 2026-02-06 DOI: 10.1016/j.talanta.2026.129520
Qingqing Pan , Jing Lin , Chunlan Tang , Tao Yu , Min Shen , Guangliang Wu , Huoyan Ji
This study developed and validated a high-throughput method based on automated magnetic solid-phase extraction (MSPE) with isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) for the simultaneous and accurate quantification of five fat-soluble vitamins (VA, 25(OH)D3, 25(OH)D2, VE, and VK1) in human serum. For the MSPE, a functionalized silica-based magnetic core-shell material was synthesized, featuring a poly (polystyrene-co-divinylbenzene-co-N-vinylpyrrolidone) coating and a covalently immobilized 2-vinylbenzofuran affinity ligand linked via a hydrophilic polyethylene glycol spacer. This design facilitated efficient enrichment and purification of the target analytes. The validated method demonstrated excellent specificity, with complete chromatographic separation, without significant matrix effects or interferences. It exhibited superior sensitivity and linearity, with a satisfactory linear response (R2 > 0.995 for all analytes) and a low limit of quantification (0.15 ng/mL-0.49 μg/mL). Recoveries were between 90.48 % and 101.95 %, while precision was satisfactory with intra-day and inter-day coefficients of variation (CV) below 7.14 % and 4.68 %, respectively. The accuracy of the method was further demonstrated by analyzing NIST standard reference material SRM 968f and SRM 972a, with a mean bias within ±7 %. Compared to traditional liquid-liquid extraction (LLE) process, the automated MSPE process efficiently extracted the target analytes from serum within 11 min through systematic optimization. Furthermore, method comparison showed a good agreement between the clinical routine LLE method and the MSPE method established. In addition, the method performed well in external quality assessment schemes, also confirming high accuracy and reliability. This robust methodology provides a powerful analytical tool for the precise monitoring of fat-soluble vitamins and nutritional status assessment in clinical practice and has the potential to promote the development of laboratory automation.
本研究建立并验证了一种基于自动磁固相萃取(MSPE) -同位素稀释液相色谱-串联质谱(ID-LC-MS/MS)的高通量方法,用于同时准确定量人血清中5种脂溶性维生素(VA、25(OH)D3、25(OH)D2、VE和VK1)。对于MSPE,合成了一种功能化硅基磁性核壳材料,该材料具有聚(聚苯乙烯-共二乙烯基苯-共n -乙烯基吡咯烷酮)涂层和通过亲水性聚乙二醇隔离剂连接的共价固定的2-乙烯基苯并呋喃亲和配体。这种设计促进了目标分析物的高效富集和纯化。经验证的方法具有良好的特异性,色谱分离完全,无明显的基质效应或干扰。该方法具有良好的灵敏度和线性,线性响应良好(所有分析物R2 >; 0.995),定量下限为0.15 ng/mL ~ 0.49 μg/mL。加样回收率在90.48% ~ 101.95%之间,精密度满意,日内变异系数(CV)分别小于7.14%和4.68%。通过分析NIST标准物质SRM 968f和SRM 972a进一步验证了该方法的准确性,平均偏差在±7%以内。与传统的液-液萃取(LLE)工艺相比,通过系统优化,自动化MSPE工艺在11 min内有效地从血清中提取出目标分析物。方法比较表明,临床常规LLE方法与建立的MSPE方法吻合较好。此外,该方法在外部质量评价方案中表现良好,也证实了较高的准确性和可靠性。这种稳健的方法为临床实践中脂溶性维生素的精确监测和营养状况评估提供了强大的分析工具,并有可能促进实验室自动化的发展。
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引用次数: 0
Chemometrics-assisted validation of a high-throughput method for quantitative analysis of multiclass phytohormones across plant matrices by UHPLC-QQQ-MS/MS for targeted metabolomics UHPLC-QQQ-MS/MS靶向代谢组学高通量定量分析植物基质中多类植物激素方法的化学计量学辅助验证
IF 6.1 1区 化学 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2026-07-01 Epub Date: 2026-02-12 DOI: 10.1016/j.talanta.2026.129541
Tingting Tong , Lili He , Liya Qiao , Yaqin Zhou , Jieqiong Li , Jie Liang , Jine Fu , Shugen Wei , Gaowu Huang , Xianghua Xia
The quantification of phytohormones in plant samples is significantly challenging because of their trace concentrations and the complex plant matrix. This study establishes and optimizes a simple and fast method for simultaneous quantification of multiclass phytohormones using ultra high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UHPLC-QQQ-MS) for high-throughput targeted metabolomics analysis. It is characterized by simplicity and operational feasibility, and it involves sequential steps of ultrasound-assisted extraction, centrifugation, and nitrogen flow-drying. The assessment of sample preprocessing method employs the technique for order preference by similarity to an ideal solution (TOPSIS) chemometrics-assisted approach, and methanol containing 1% formic acid was selected as the optimal extraction solvent. Quantitative analysis is performed via a multiple reaction monitoring method. The method was validated satisfactory for 31 phytohormones, the specific concentration levels used to estimate recovery and precision were 0.25–200 ng/mL, and the results showed good linearity (correlation coefficients: 0.9910–0.9999), sensitivity (LOD: 0.01–2.87 ng/g; LOQ: 0.02–8.70 ng/g), accuracy (recovery rates: 64.50–129.67%), precision (relative standard deviations <20%, except for IAA-Trp), and its applicability to a variety species of medicinal plant was verified, including Taxillus chinensis (DC.) Danser, Ophiopogon japonicus (L.f.) Ker-Gawl, Citrus grandis “Tomentosa,” Plumeria rubra L. “Acutifolia,” and Pogostemon cablin (Blanco) Benth. In the real samples analyzed, auxins concentrations exceeded 4.07 μg/g, salicylic acids concentrations ranged from 0.027 to 79.7 μg/g, and concentrations of other categorize of phytohormones were below 1 μg/g. The established method demonstrated high throughput, accuracy, stability, and reliability. It can be extended to cover additional phytohormone types and diverse application scenarios. This method serves as a potent tool for investigating the function of multiclass phytohormones in the regulation of secondary metabolite accumulation in medicinal plants, elucidating complex signal transduction mechanisms and interaction networks, and laying the groundwork for improving the quality of Chinese medicinal materials.
由于植物样品中植物激素的痕量浓度和复杂的植物基质,对植物激素的定量分析具有很大的挑战性。本研究建立并优化了一种高效液相色谱-三重四极杆串联质谱(UHPLC-QQQ-MS)同时定量多类植物激素的方法,用于高通量靶向代谢组学分析。它的特点是简单和操作可行性,它涉及超声辅助提取,离心和氮流干燥的顺序步骤。样品预处理方法的评价采用TOPSIS化学计量辅助相似性排序法,选择含1%甲酸的甲醇作为最佳提取溶剂。定量分析是通过多反应监测方法进行的。该方法对31种植物激素的检测结果满意,测定回收率和精密度范围为0.25 ~ 200 ng/mL,线性良好(相关系数为0.9910 ~ 0.9999),灵敏度为0.01 ~ 2.87 ng/g;LOQ: 0.02 ~ 8.70 ng/g)、准确度(回收率:64.50 ~ 129.67%)、精密度(相对标准偏差:20%,除iaa -色氨酸外),并验证了其对多种药用植物的适用性,包括紫杉(Taxillus chinensis, DC)。丹瑟,日本麦冬(L.f)大柑橘“Tomentosa”,鸡蛋花L.“Acutifolia”和广藿香(Blanco) Benth。在实际样品中,生长素浓度超过4.07 μg/g,水杨酸浓度在0.027 ~ 79.7 μg/g之间,其他类型的植物激素浓度在1 μg/g以下。所建立的方法具有较高的通量、准确度、稳定性和可靠性。它可以扩展到涵盖更多的植物激素类型和不同的应用场景。该方法可为研究多类植物激素调控药用植物次生代谢物积累的功能,阐明复杂的信号转导机制和相互作用网络,为提高中药材质量奠定基础。
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引用次数: 0
A sensitive pH fluorescent probe with large Stokes shift and narrow transition for lysosome imaging during cell cycle 一种敏感的pH荧光探针,具有大斯托克斯位移和窄跃迁,用于细胞周期溶酶体成像
IF 6.1 1区 化学 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2026-07-01 Epub Date: 2026-02-10 DOI: 10.1016/j.talanta.2026.129534
Bochao Chen , Jixiang Zhang , Yifan Zhong , Xi Liu , Ji Xu , Ningning Cui , Jin Zhou
Intracellular pH plays a crucial role in regulating cellular functions, and its subtle changes can influence normal physiological activities and contribute to various diseases. Although numerous probes have been developed for detecting microenvironmental pH in living cells, their broad dynamic ranges induce blurred imaging boundaries and may generate false-positive results, compromising the accurate detection of subtle pH fluctuations. In response to these issues, we designed and synthesized a novel 1,8-naphthalic anhydride-based pH fluorescent probe (NAM) that demonstrated highly sensitive responses to pH changes with a narrow transition range spanning just 2.4 pH units. The pH responsiveness of NAM originated from the deprotonation of its aniline group, which induced an enhanced intramolecular charge transfer (ICT) process. This mechanism enables NAM to exhibit a remarkable 15-fold fluorescence enhancement at 569 nm with a pKa value of 6.71 and a significant Stokes shift of 112 nm. Furthermore, NAM specifically targets lysosomes and was successfully employed to detect intracellular pH fluctuations induced by lipopolysaccharide (LPS), nutrient deprivation and chloroquine treatment. Significantly, we demonstrate for the first time that NAM achieved pH imaging analysis across different cell cycle phases. During the transition from G0/G1 to S phase, intracellular pH gradually alkalinizes, followed by rapid acidification in the G2/M phase. These observations reflected the dynamic balance between energy storage and consumption during cell cycle progression. These distinctive features position NAM as a promising tool for investigating physiological and pathological processes associated with abnormal pH variations.
细胞内pH值在调节细胞功能中起着至关重要的作用,其细微的变化会影响正常的生理活动,导致多种疾病的发生。尽管已经开发了许多探针用于检测活细胞中的微环境pH值,但它们的宽动态范围会导致成像边界模糊,并可能产生假阳性结果,从而影响对细微pH值波动的准确检测。针对这些问题,我们设计并合成了一种新型的1,8-萘酸酐基pH荧光探针(NAM),该探针对pH变化具有高度敏感的响应,其过渡范围仅为2.4 pH单位。NAM的pH响应性源于其苯胺基的去质子化,这引起了分子内电荷转移(ICT)过程的增强。这种机制使得NAM在569 nm处表现出15倍的荧光增强,pKa值为6.71,Stokes位移为112 nm。此外,NAM特异性靶向溶酶体,并成功用于检测由脂多糖(LPS)、营养剥夺和氯喹处理引起的细胞内pH波动。值得注意的是,我们首次证明了NAM在不同细胞周期阶段实现了pH成像分析。在G0/G1期向S期过渡期间,细胞内pH逐渐碱化,随后在G2/M期快速酸化。这些观察结果反映了细胞周期进程中能量储存和消耗之间的动态平衡。这些独特的特征使NAM成为研究与异常pH变化相关的生理和病理过程的有前途的工具。
{"title":"A sensitive pH fluorescent probe with large Stokes shift and narrow transition for lysosome imaging during cell cycle","authors":"Bochao Chen ,&nbsp;Jixiang Zhang ,&nbsp;Yifan Zhong ,&nbsp;Xi Liu ,&nbsp;Ji Xu ,&nbsp;Ningning Cui ,&nbsp;Jin Zhou","doi":"10.1016/j.talanta.2026.129534","DOIUrl":"10.1016/j.talanta.2026.129534","url":null,"abstract":"<div><div>Intracellular pH plays a crucial role in regulating cellular functions, and its subtle changes can influence normal physiological activities and contribute to various diseases. Although numerous probes have been developed for detecting microenvironmental pH in living cells, their broad dynamic ranges induce blurred imaging boundaries and may generate false-positive results, compromising the accurate detection of subtle pH fluctuations. In response to these issues, we designed and synthesized a novel 1,8-naphthalic anhydride-based pH fluorescent probe (<strong>NAM</strong>) that demonstrated highly sensitive responses to pH changes with a narrow transition range spanning just 2.4 pH units. The pH responsiveness of <strong>NAM</strong> originated from the deprotonation of its aniline group, which induced an enhanced intramolecular charge transfer (ICT) process. This mechanism enables <strong>NAM</strong> to exhibit a remarkable 15-fold fluorescence enhancement at 569 nm with a pKa value of 6.71 and a significant Stokes shift of 112 nm. Furthermore, <strong>NAM</strong> specifically targets lysosomes and was successfully employed to detect intracellular pH fluctuations induced by lipopolysaccharide (LPS), nutrient deprivation and chloroquine treatment. Significantly, we demonstrate for the first time that <strong>NAM</strong> achieved pH imaging analysis across different cell cycle phases. During the transition from G0/G1 to S phase, intracellular pH gradually alkalinizes, followed by rapid acidification in the G2/M phase. These observations reflected the dynamic balance between energy storage and consumption during cell cycle progression. These distinctive features position <strong>NAM</strong> as a promising tool for investigating physiological and pathological processes associated with abnormal pH variations.</div></div>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"304 ","pages":"Article 129534"},"PeriodicalIF":6.1,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146187179","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
HRP-induced wide wavelength-tunable in situ fluorogenic system and its immunoassay for cTnI in clinical samples hrp诱导的宽波长可调原位荧光系统及其临床样品中cTnI的免疫测定
IF 6.1 1区 化学 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2026-07-01 Epub Date: 2026-02-05 DOI: 10.1016/j.talanta.2026.129511
Jingjing Yin , Yuehua Wang , Shanshan Zhu , Xiaoxue Liu, Yujie Sun, Hong Yang, Yifei Ma, Jinhua Liu
The development of wavelength-tunable fluorescence immunoassays for sensitive biomarker detection remains a significant challenge in clinical diagnostics. In this work, we present an HRP-induced in situ fluorogenic immunoassay platform based on the rapid reaction between p-phenylenediamine (PPD) analogues and 6-hydroxyquinoline (6-Hy). The proposed mechanism involves HRP-catalyzed generation of hydroxyl radicals from H2O2, which subsequently oxidize PPD to form fluorescent copolymer nanoparticles (FCNPs) with 6-Hy. By strategically modifying the PPD structure with various substituents, we achieved discrete and tunable emission of FCNPs, with maxima spanning from 380 to 600 nm (Δλ ≈ 220 nm). Using cardiac troponin I (cTnI) as a model antigen and zinc-coordination nanoparticles (ZnNPs) as carriers, the immunoassay demonstrated exceptional performance, with a wide dynamic range (0.5-125 ng/mL) and an ultralow detection limit (0.17 ng/mL) for cTnI. The platform was successfully validated in human serum samples, showing excellent correlation with clinical standards. This work not only establishes a novel strategy for wavelength-programmable immunoassays but also provides a versatile platform for advancing biomarker detection in clinical diagnostics.
开发波长可调的荧光免疫检测技术用于灵敏的生物标志物检测仍然是临床诊断中的一个重大挑战。在这项工作中,我们建立了一个基于对苯二胺(PPD)类似物和6-羟基喹啉(6-Hy)之间快速反应的酶促原位荧光免疫分析平台。所提出的机制涉及hrp催化H2O2产生羟基自由基,随后氧化PPD形成具有6-Hy的荧光共聚物纳米颗粒(FCNPs)。通过用不同的取代基修饰PPD结构,我们实现了FCNPs的离散和可调谐发射,最大发射距离为380 ~ 600 nm (Δλ≈220 nm)。以心肌肌钙蛋白I (cTnI)为模型抗原,锌配位纳米颗粒(ZnNPs)为载体,具有较宽的动态范围(0.5 ~ 125 ng/mL)和超低的检测限(0.17 ng/mL)。该平台已在人血清样本中成功验证,与临床标准具有良好的相关性。这项工作不仅建立了波长可编程免疫测定的新策略,而且为推进临床诊断中的生物标志物检测提供了一个通用的平台。
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引用次数: 0
Extended-gate FET biosensor utilizing ionic strength engineering for sensitive prostate-specific antigen detection 利用离子强度工程进行前列腺特异性抗原检测的扩展栅场效应晶体管生物传感器
IF 6.1 1区 化学 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2026-07-01 Epub Date: 2026-02-14 DOI: 10.1016/j.talanta.2026.129531
Sheng-Chun Hung , Kai-Chun Hung , Chia Kai Lin
This work presents an extended-gate field-effect transistor (EGFET) biosensor functionalized with prostate-specific antigen (PSA) aptamers. It employs ionic strength engineering to enhance electrostatic coupling and improve detection sensitivity. By systematically adjusting the concentration of phosphate-buffered saline (PBS), we modified the electric double-layer (EDL) structure and Debye length to investigate their effects on charge screening and signal transduction. The proposed biosensor achieved a detection limit of 0.408 ng/mL in 0.001 × PBS, which is significantly lower than the clinical threshold of 4 ng/mL. Additionally, it exhibited an impressive nearly fivefold increase in sensitivity as the ionic strength decreased from 1 × to 0.001 × PBS. These enhancements can be attributed to extended Debye screening and reduced electrostatic shielding in low-ionic environments, facilitating more efficient field-effect coupling between the aptamer–PSA complex and the transistor channel. Temporal noise characterization, conducted through Allan deviation analysis, revealed an optimal averaging range of 30 to 80 s. This range effectively balances noise suppression and signal fidelity. In summary, this study demonstrates a straightforward and effective strategy for regulating ionic strength to enhance the transduction efficiency of BioFET-based biosensors, offering a practical route toward high-performance and label-free PSA biosensing in biomedical diagnostics.
这项工作提出了一个扩展门场效应晶体管(EGFET)生物传感器功能与前列腺特异性抗原(PSA)适配体。采用离子强度工程增强静电耦合,提高检测灵敏度。通过系统调节磷酸盐缓冲盐水(PBS)的浓度,我们改变了双电层(EDL)结构和德拜长度,研究了它们对电荷筛选和信号转导的影响。该生物传感器在0.001 × PBS中检测限为0.408 ng/mL,明显低于临床阈值4 ng/mL。此外,当离子强度从1 × PBS降低到0.001 × PBS时,它的灵敏度增加了近5倍。这些增强可归因于低离子环境中扩展的德拜筛选和减少的静电屏蔽,促进适体- psa复合物与晶体管通道之间更有效的场效应耦合。通过Allan偏差分析进行的时间噪声表征显示,最佳平均范围为30至80秒。这个范围有效地平衡了噪声抑制和信号保真度。总之,本研究展示了一种简单有效的策略来调节离子强度以提高基于biofet的生物传感器的转导效率,为生物医学诊断中高性能和无标记的PSA生物传感提供了一条实用的途径。
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引用次数: 0
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