Analytical and Bioanalytical Chemistry最新文献

{"title":"Ratiometric photoelectrochemical sensors: advances in food analysis and emerging applications.","authors":"Yuanqiang Hao, Jinting Sun, Peisheng Zhang, Rongjin Zeng, Maotian Xu, Shu Chen","doi":"10.1007/s00216-025-06102-9","DOIUrl":"https://doi.org/10.1007/s00216-025-06102-9","url":null,"abstract":"<p><p>Ratiometric photoelectrochemical (PEC) sensors have emerged as a compelling class of analytical tools, offering internal calibration, enhanced signal stability, and improved tolerance to environmental variability. These advantages are particularly relevant to the field of food analysis, where sample complexity often compromises the reliability of single-signal detection strategies. In recent years, diverse ratiometric PEC platforms have been developed. This review systematically categorizes ratiometric PEC sensors into five main types: wavelength-resolved, voltage-resolved, spatially resolved, dual-mode, and other emerging strategies involving polarity switching or stimulus-responsive interfaces. For each category, representative sensing mechanisms, photoelectrode architectures, and real-world applications-especially in the detection of food contaminants such as mycotoxins, antibiotics, and pesticide residues-are comprehensively discussed. Furthermore, key challenges and future directions are outlined. This review aims to provide a conceptual foundation and technical reference for advancing ratiometric PEC sensing technologies in food safety and beyond.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145068854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"WSe₂ nanosheets with peroxidase-mimetic activity and carbon dots based ratiometric fluorescent strategy for D-galactose sensing.","authors":"Sikai Wang, Weiping Liu, Lei Wang, Lin Lv, Xiangqin Liu, Xiaojun Luo, Guoqi Zhang, Yan Zhao","doi":"10.1007/s00216-025-06104-7","DOIUrl":"https://doi.org/10.1007/s00216-025-06104-7","url":null,"abstract":"<p><p>D-Galactose (D-Gal), an essential monosaccharide, plays multiple physiological roles in living organisms. Abnormal fluctuations in its concentration are closely associated with various genetic metabolic disorders, such as galactosemia and galactose deficiency. Therefore, the development of an efficient, sensitive, and accurate method for D-Gal detection is critical for disease diagnosis and treatment. In this study, a ratiometric fluorescence-based sensing platform was constructed for the quantitative detection of D-Gal. Galactose oxidase can catalyze the oxidation of D-Gal, resulting in the generation of hydrogen peroxide (H₂O₂). When combined with WSe₂ nanosheets prepared via liquid-phase exfoliation and functioning as nanozymes, in the presence of H₂O₂, the substrate o-phenylenediamine (OPD) undergoes oxidation to generate 2,3-diaminophenazine (DAP), which exhibits a fluorescence peak at 570 nm. Simultaneously, DAP quenches the fluorescence of carbon dots (CDs) at 430 nm via an inner filter effect, thereby enabling ratiometric detection with enhanced accuracy and sensitivity. The method exhibits a Limit of detection of 0.004 mM, Linear ranges spanning from 0.01 to 1 mM and from 1 to 300 mM, with recoveries ranging between 91.0 and 100.4%. Furthermore, the proposed method was effectively utilized for the detection of D-Gal in human serum specimens, demonstrating excellent selectivity and reliability. This research presents a novel method for D-Gal detection based on WSe₂ nanosheets and CDs, offering potential applications in the biomedical and biosensing fields.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145068859","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Challenges and advances in analytical separation techniques for RNA-lipid nanoparticle therapeutics.","authors":"Brady W Drennan, Kevin A Schug","doi":"10.1007/s00216-025-06096-4","DOIUrl":"https://doi.org/10.1007/s00216-025-06096-4","url":null,"abstract":"<p><p>Gene therapies are rapidly advancing as drug modalities, serving as a means to treat previously undruggable pathways. The use of small interfering ribonucleic acid (siRNA), messenger RNA (mRNA), and guide RNA (gRNA) in clustered regularly interspaced short palindromic repeats (CRISPR)-based gene editing, combined with lipid nanoparticles (LNPs), has demonstrated effective drug delivery. These complex delivery systems often require multiple analytical methodologies to achieve comprehensive characterization, some of which remain underdeveloped or inadequately adapted for RNA-LNP formulations. Commonly used batch-based methods, such as dynamic light scattering (DLS) or the modified RiboGreen assay, are frequently hindered by sample heterogeneity, a limitation that can be addressed through analytical separations. This review discusses the challenges limiting analytical separations for RNA-LNP therapeutics and highlights recent advances in separation science for reliable characterization and quality control. We focus on techniques for RNA, LNPs, and the RNA-LNP complex, emphasizing chromatographic, electrophoretic, and field-based separation techniques.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145051562","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A new wide-scope, multi-biomarker wastewater-based epidemiology analytical method to monitor the health and well-being of inhabitants at a metropolitan scale","authors":"Harry Elliss,&nbsp;Kit Proctor,&nbsp;Megan Robertson,&nbsp;John Bagnall,&nbsp;Barbara Kasprzyk-Hordern","doi":"10.1007/s00216-025-06097-3","DOIUrl":"10.1007/s00216-025-06097-3","url":null,"abstract":"<div><p>This manuscript establishes a new, comprehensive biomarker list and a multiresidue trace quantification method for community-wide health and well-being assessment at a metropolitan scale using wastewater-based epidemiology (WBE) and mass spectrometry pipelines. This method enables the quantification of 204 biochemical indicators (BCIs) across a range of biomarker classes within influent wastewater and includes illicit drug BCIs, pharmaceuticals as proxies for disease, health markers (hormones, oxidative stress, lipid peroxidation, etc.), Lifestyle chemicals, food BCIs, and hazardous chemicals in personal care products. This method facilitates the combined assessment of community exposure to chemicals and the effects of this exposure in the same framework. The method enables full quantification of 141 BCIs with method detection Limits varying from 0.01 ng/L for amlodipine to 23.8 ng/L for stachydrine. Total average method accuracies were 102.7% whereas precision was 10.4%. During an initial assessment of this method to test its suitability, 62% of all targets were detected and quantified during a week-long feasibility study of a large city with weekly average Daily BCI loads ranging from 40.0 ± 20.0 mg/day for salbutamol to 5836.5 ± 1697.1 g/day for creatinine. The inclusion of new endogenous markers such as advanced glycation end products, detected in wastewater for the first time, enables more accurate determination of community-level health and lifestyle habits. Alongside an unbiased and comprehensive health assessment through endogenous markers, health is further assessed via the use of pharmaceuticals, acting as a proxy for health and disease status whilst additionally providing insights into community lifestyle habits through the monitoring of licit/illicit drug use and food consumption. The analysis of all biomarker classes combined aims to provide insights to exposure and health effect outcomes at the community level.</p><h3>Graphical Abstract</h3>\u0000<div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":"417 26","pages":"5983 - 6005"},"PeriodicalIF":3.8,"publicationDate":"2025-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00216-025-06097-3.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145051493","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biomarker discovery for lung adenocarcinoma diagnosis using liquid chromatography-mass spectrometry-based enhanced pseudotargeted metabolomics.","authors":"Guoqin Ji, Di Yu, Luhan Li, Jinhui Zhao, Xiaolin Wang, Siqi Zhu, Shiheng Luo, Xiaodong Li, Guowang Xu, Penglong Cao, Xinyu Liu","doi":"10.1007/s00216-025-06092-8","DOIUrl":"https://doi.org/10.1007/s00216-025-06092-8","url":null,"abstract":"<p><p>Lung adenocarcinoma, the most prevalent subtype of non-small cell lung cancer, is often diagnosed at advanced stages due to the lack of effective early screening methods, leading to poor patient outcomes. In this study, an enhanced pseudotargeted metabolomics approach was developed using liquid chromatography-mass spectrometry, combining untargeted-level coverage with targeted quantitative accuracy while enabling simplified clinical implementation. Serum samples from early-stage lung adenocarcinoma (LUAD) patients and healthy controls were analyzed using this method to identify potential biomarkers and establish a diagnostic model for early LUAD detection. A total of 329 serum samples were divided into discovery, internal validation, and external validation cohorts. Through non-parametric tests and machine learning algorithms, 113 differential metabolites were identified. Glycerophosphocholine and glutamine were validated as potential biomarkers for early LUAD diagnosis; the diagnostic model based on these biomarkers demonstrated good discriminative power, with AUCs of 0.972 and 0.867 in the internal and external validations, respectively. Additionally, comparative analysis between stage I and stage II patients revealed significant metabolic changes including elevated levels of choline, and sphingosine, and decreased levels of 3-dehydroteasterone and PC 31:0. These findings provided new insights into the metabolic alterations associated with LUAD progression and highlighted the potential of pseudotargeted metabolomics in discovering the metabolite biomarkers for early diagnosis of LUAD.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145038876","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of smartphone-based AIE fluorescence-quenching immunochromatographic sensors for the detection of illicit drugs in various complex sample matrices.","authors":"Lijiao Liu, Guanbo He, Lifang Ye, Yong He, Tao Xu, Xiaoli Zhang, Chenxing Zeng, Qifang Song, Jiajie Liang, Yong Tang","doi":"10.1007/s00216-025-06106-5","DOIUrl":"https://doi.org/10.1007/s00216-025-06106-5","url":null,"abstract":"<p><p>Illicit drug abuse poses a significant global threat to public health and social security, highlighting the urgent need for rapid, sensitive, and versatile detection technologies. To address the limitations of traditional chromatographic techniques-such as high costs and slow response times-and the drawbacks of conventional immunochromatographic sensors (ICS), including low sensitivity and non-intuitive signal outputs, a fluorescence-quenching ICS (FQICS) was developed. This sensor leverages fluorescence resonance energy transfer (FRET) between aggregation-induced emission fluorescent microspheres (AIEFMs) and gold nanoparticles (AuNPs). The ICS operates on a positive signal-readout mechanism and is integrated with a smartphone-based portable reader, enabling rapid quantitative detection of methamphetamine (MET), morphine (MOR), and ketamine (KET). Detection limits of 0.041, 0.072, and 0.059 ng mL^-1 were determined for MET, MOR, and KET, respectively. Recovery rates ranged from 73% to 134% across urine, hair, saliva, and sewage samples, with intra-assay precision consistently below 15%, indicating robust performance in complex matrices. Furthermore, we developed a multiplexed AIEFM-FQICS, enabling the simultaneous detection of three illicit drugs, thereby enhancing detection efficiency and reducing the cost. Generally, this work presents a highly sensitive, field-deployable platform for real-time monitoring in drug interdiction and public safety emergencies, offering substantial potential for practical anti-drug applications.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145032556","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ultrasensitive and specific electrochemical detection of Escherichia coli via functionalized magnetic nanoparticles by time–frequency analysis","authors":"Siqi Dong,&nbsp;Xiaobin Zhang,&nbsp;Shijuan Cao,&nbsp;Chenpan Lei,&nbsp;Hanyang Bao,&nbsp;Ying Xu","doi":"10.1007/s00216-025-06059-9","DOIUrl":"10.1007/s00216-025-06059-9","url":null,"abstract":"<div><p>The prompt and accurate identification of pathogenic bacteria is crucial for mitigating the transmission of infections. Conventional detection methods face limitations, including lengthy processing, complex sample pretreatment, high instrumentation costs, and insufficient sensitivity for rapid on-site screening. To address these challenges, an aptamer (Apt)-sensor based on functionalized magnetic nanoparticles (MNPs) was developed for detecting <i>Escherichia coli</i>. Fe₃O₄@Au nanoparticles were synthesized by stepwise modification, followed by Apt conjugation via Au–S bonds to form Fe₃O₄@Au@Apt. Subsequently, efficient capture and separation of target bacteria was achieved by combining the specific Apt-<i>E. coli</i> recognition sites with magnetic solid-phase extraction. A time–frequency domain feature-assisted XGBoost model was constructed for the sensor to achieve accurate prediction of bacterial concentration. Six equivalent-circuit frequency domain and six time domain characteristic parameters were extracted from the equivalent circuit model (ECM) and the distribution of relaxation times (DRT), respectively, and Bayesian optimization (BO) was subsequently adopted for automatic hyperparameter search to reduce prediction errors. Furthermore, the SHapley Additive exPlanations (SHAP) analysis demonstrated the necessity of time–frequency feature fusion for enhancing prediction accuracy. The experimental results indicated that the Fe₃O₄@Au@Apt-modified magnetic glassy carbon electrode (MGCE) can achieve quantitative detection of <i>E. coli</i> in a concentration range of 10⁰–10⁷ CFU/mL, with a detection Limit down to 1 CFU/mL. In addition, the intelligent detection framework based on BO-XGBoost exhibited excellent predictive performance, with an <i>R</i>² value of 0.990, mean absolute error (MAE) of 0.087 CFU/mL, and root mean square error (RMSE) of 0.158 CFU/mL. This approach shows significant potential for future <i>E. coli</i> monitoring applications in food safety and environmental surveillance.</p></div>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":"417 24","pages":"5449 - 5461"},"PeriodicalIF":3.8,"publicationDate":"2025-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145032619","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Carbon quantum dot-aptamer/MoS2 nanosheet fluorescent sensor for ultrasensitive, noninvasive cortisol detection","authors":"Rong Chen,&nbsp;Mingyu Wang,&nbsp;Hailiang Nie,&nbsp;Jinku Zhang,&nbsp;Hongyuan Yan","doi":"10.1007/s00216-025-06086-6","DOIUrl":"10.1007/s00216-025-06086-6","url":null,"abstract":"<div><p>This work presents the development of a highly sensitive, selective, and efficient aptamer-based fluorescent sensor for detecting cortisol in human urine. Carbon quantum dots-nucleic acid aptamer (CQDs-Apt) synthesized with excellent photoluminescent properties and stability, were selected as the fluorescent probe. In the presence of MoS₂-NSs, CQDs-Apt adsorbed onto the surface of MoS₂-NSs via electrostatic and π-π interactions, leading to strong and rapid fluorescence quenching due to static quenching mechanism between them. The CQDs-Apt/MoS₂-NSs complex can be employed as a “turn-on” fluorescent sensor for cortisol. Upon the addition of cortisol to the CQDs-Apt/MoS₂-NSs sensor, the aptamer specifically binds to cortisol, thereby weakening the interaction between CQDs-Apt and MoS₂-NSs. This results in the desorption of CDs-Apt from the surface of MoS₂-NSs and the recovery of the fluorescence signal. Under optimized conditions, the CQDs-Apt/MoS₂-NSs sensor exhibits a linear response to cortisol concentration (1–500 ng/mL) with a detection limit of 0.3 ng/mL. Furthermore, the sensor demonstrated excellent stability, high accuracy (92.0% to 97.7%), and superior precision (RSD ≤ 3.5%). This sensor has achieved a sensitive, rapid and efficient response to cortisol, and been successfully utilized for the detection of cortisol in human urine, demonstrating its potential for clinical cortisol detection.</p></div>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":"417 26","pages":"5919 - 5930"},"PeriodicalIF":3.8,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145022608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a certified reference material for per- and polyfluoroalkyl substances (PFAS) in textiles","authors":"Thomas Sommerfeld,&nbsp;Juliane Riedel,&nbsp;Jan Lisec,&nbsp;Tatjana Mauch,&nbsp;Silke Richter,&nbsp;Matthias Koch","doi":"10.1007/s00216-025-06098-2","DOIUrl":"10.1007/s00216-025-06098-2","url":null,"abstract":"<div><p>Per- and polyfluoroalkyl substances (PFASs) are a large group of emerging organic pollutants that contaminate the environment, food, and consumer products. Textiles and other outdoor products are a major source of PFAS exposure due to their water-repellent impregnations. Determination of PFASs in textiles is increasingly important for enhancing their contribution to the circular economy. While maximum levels and restrictions exist for certain key compounds under the Stockholm Convention on Persistent Organic Pollutants and the REACH regulation, certified reference materials (CRMs) are not currently available. To address this issue, the first CRM for determining PFASs in outdoor textiles (BAM-B003) was developed. It fully complies with the requirements of ISO 17034 and ISO 33405. This work presents the entire process of CRM development process, including preparation, a homogeneity study, a stability study, and value assignment. Certification was based on an in-house study at BAM using liquid chromatography tandem mass spectrometry (LC–MS/MS) with stable isotope dilution analysis (SIDA). The certified mass fractions of 18 PFASs range widely from 0.46 to 69 µg/kg, with a prevalence of PFOA (69 µg/kg), PFOS (41 µg/kg) and PFHxA (35 µg/kg) exceeding legal limits. BAM-B003 is intended for analytical quality control and contributes to improving the chemical safety of textiles and strengthening the circular economy.</p><h3>Graphical Abstract</h3>\u0000<div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":"417 26","pages":"6007 - 6015"},"PeriodicalIF":3.8,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00216-025-06098-2.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145028783","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Asymmetric volume-mediated buffer control overcomes sensitivity limits in one-pot RAA-CRISPR/Cas12a visual detection","authors":"Yue Zhang,&nbsp;Zunquan Zhao,&nbsp;Mingzhu Liu,&nbsp;Jincai Yang,&nbsp;Chun Yang,&nbsp;Nan Su,&nbsp;Jingran Sun,&nbsp;Yanjun Fang,&nbsp;Yonghui Wang,&nbsp;Xiaoli Li,&nbsp;Wang Chen,&nbsp;Jin Wu,&nbsp;Jialei Bai","doi":"10.1007/s00216-025-06095-5","DOIUrl":"10.1007/s00216-025-06095-5","url":null,"abstract":"<div><p>Rapid, low-cost, and visual nucleic acid detection methods are highly attractive for curbing colistin resistance spread through the food chain. CRISPR/Cas12a combined with recombinase-aided amplification (RAA) offers a one-pot, aerosol-free approach for visual detection. However, traditional one-pot systems often run Cas12a <i>trans</i>-cleavage in a buffer suitable for RAA, thus limiting Cas12a cleavage efficiency. This study proposes an asymmetric volume-optimized RAA-CRISPR/Cas12a assay for ultrasensitive visual detection of mobile colistin resistance gene <i>mcr-1</i>. Unlike conventional one-pot systems constrained by buffer incompatibility, our design spatially segregates a minimal-volume RAA-MIX (lid) from a CRISPR-dominant buffer microenvironment (tube bottom). This architecture leverages RAA’s exponential amplification power to ensure sufficient product yield from minimal reaction volumes, while enabling subsequent enhancement of Cas12a <i>trans</i>-cleavage through automatic buffer assimilation upon mixing. The results were able to be visually observed under UV light, achieving 63.1% cost reduction compared to standard one-pot methods. The sensitivity of the proposed method for the <i>mcr-1</i> gene was 2.5 copies/reaction, with anti-interference against other plasmids or bacteria. This method was applied to the detection of <i>mcr-1</i> in animal-derived foods, showing satisfactory practical performance. By fundamentally reengineering buffer microenvironments through volume asymmetry, this work provides a general strategy for one-pot molecular diagnostics, achieving dual optimization of amplification and cleavage without trade-offs.</p><h3>Graphical Abstract</h3>\u0000<div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":"417 26","pages":"5971 - 5981"},"PeriodicalIF":3.8,"publicationDate":"2025-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145022564","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}