Pub Date : 2025-04-03DOI: 10.1021/acs.analchem.4c06431
Jiaxing Chang, Zhinan Zhang, Chulin Qu, Qingzhi Han, Li Xu
We report a strategy based on pyridyl-anchored organic small-molecule fluorescent probes to develop a dual-signal sensing platform. The strategy accomplishes an intelligent integration of fluorescence analysis with photoelectrochemical (PEC) sensing, thereby enabling rapid and precise detection of hypochlorite. In this work, the natural dye chromone was selected as the fluorophore for generating fluorescent signals. Meanwhile, by using phenothiazine (PTZ) as the specific recognition group and pyridine as the anchoring moiety, we designed and synthesized a novel organic small-molecule fluorescent probe. The obtained probe was used as a photosensitive material anchored to the TiO2 surface via N → Ti bonds, to form an FTO/TiO2/FPTZ-1 heterostructure-based dual-signal sensing platform for the detection of hypochlorite. This sensing platform has the characteristics of high specificity, sensitivity, and ease of preparation, enabling rapid qualitative fluorescence readout and quantitative photoelectrochemical readout of hypochlorite, with a limit of detection of 0.288 μM for fluorescence and 1.37 nM for PEC.
{"title":"Organic Molecules as a Bridge Connecting Photoelectrochemistry and Fluorescence for Dual-Signal Assay","authors":"Jiaxing Chang, Zhinan Zhang, Chulin Qu, Qingzhi Han, Li Xu","doi":"10.1021/acs.analchem.4c06431","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c06431","url":null,"abstract":"We report a strategy based on pyridyl-anchored organic small-molecule fluorescent probes to develop a dual-signal sensing platform. The strategy accomplishes an intelligent integration of fluorescence analysis with photoelectrochemical (PEC) sensing, thereby enabling rapid and precise detection of hypochlorite. In this work, the natural dye chromone was selected as the fluorophore for generating fluorescent signals. Meanwhile, by using phenothiazine (PTZ) as the specific recognition group and pyridine as the anchoring moiety, we designed and synthesized a novel organic small-molecule fluorescent probe. The obtained probe was used as a photosensitive material anchored to the TiO<sub>2</sub> surface via N → Ti bonds, to form an FTO/TiO<sub>2</sub>/FPTZ-1 heterostructure-based dual-signal sensing platform for the detection of hypochlorite. This sensing platform has the characteristics of high specificity, sensitivity, and ease of preparation, enabling rapid qualitative fluorescence readout and quantitative photoelectrochemical readout of hypochlorite, with a limit of detection of 0.288 μM for fluorescence and 1.37 nM for PEC.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"25 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143766648","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}
Pub Date : 2025-04-03DOI: 10.1021/acs.analchem.4c05315
Yao-Yu Chen, Na An, Yan-Zhen Wang, Peng-Cheng Mei, Jun-Di Hao, Song-Mei Liu, Quan-Fei Zhu, Yu-Qi Feng
Metabolomics, which involves the comprehensive analysis of small molecules within biological systems, plays a crucial role in elucidating the biochemical underpinnings of physiological processes and disease conditions. However, current coverage of the metabolome remains limited. In this study, we present a heuristic strategy for untargeted metabolomics analysis (HeuSMA) based on multiple chromatographic gradients to enhance the metabolome coverage in untargeted metabolomics. This strategy involves performing LC-MS analysis under multiple gradient conditions on a given sample (e.g., a pooled sample or a quality control sample) to obtain a comprehensive metabolomics data set, followed by constructing a heuristic peak list using a retention index system. Guided by this list, heuristic peak picking in quantitative metabolomics data is achieved. The benchmarking and validation results demonstrate that HeuSMA outperforms existing tools (such as MS-DIAL and MZmine) in terms of metabolite coverage and peak identification accuracy. Additionally, HeuSMA improves the accessibility of MS/MS data, thereby facilitating the metabolite annotation. The effectiveness of the HeuSMA strategy was further demonstrated through its application in serum metabolomics analysis of human hepatocellular carcinoma (HCC). To facilitate the adoption of the HeuSMA strategy, we also developed two user-friendly graphical interface software solutions (HPLG and HP), which automate the analysis process, enabling researchers to efficiently manage data and derive meaningful conclusions (https://github.com/Lacterd/HeuSMA).
{"title":"HeuSMA: A Multigradient LC-MS Strategy for Improving Peak Identification in Untargeted Metabolomics","authors":"Yao-Yu Chen, Na An, Yan-Zhen Wang, Peng-Cheng Mei, Jun-Di Hao, Song-Mei Liu, Quan-Fei Zhu, Yu-Qi Feng","doi":"10.1021/acs.analchem.4c05315","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c05315","url":null,"abstract":"Metabolomics, which involves the comprehensive analysis of small molecules within biological systems, plays a crucial role in elucidating the biochemical underpinnings of physiological processes and disease conditions. However, current coverage of the metabolome remains limited. In this study, we present a heuristic strategy for untargeted metabolomics analysis (HeuSMA) based on multiple chromatographic gradients to enhance the metabolome coverage in untargeted metabolomics. This strategy involves performing LC-MS analysis under multiple gradient conditions on a given sample (e.g., a pooled sample or a quality control sample) to obtain a comprehensive metabolomics data set, followed by constructing a heuristic peak list using a retention index system. Guided by this list, heuristic peak picking in quantitative metabolomics data is achieved. The benchmarking and validation results demonstrate that HeuSMA outperforms existing tools (such as MS-DIAL and MZmine) in terms of metabolite coverage and peak identification accuracy. Additionally, HeuSMA improves the accessibility of MS/MS data, thereby facilitating the metabolite annotation. The effectiveness of the HeuSMA strategy was further demonstrated through its application in serum metabolomics analysis of human hepatocellular carcinoma (HCC). To facilitate the adoption of the HeuSMA strategy, we also developed two user-friendly graphical interface software solutions (HPLG and HP), which automate the analysis process, enabling researchers to efficiently manage data and derive meaningful conclusions (https://github.com/Lacterd/HeuSMA).","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"11 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143766646","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}
The versatile fluorescent dyes are essential for specifically labeling plant cell walls in vivo, monitoring plasma membrane damage, and assessing cell viability. However, such dyes are rare and often discovered accidentally due to a lack of design principles. Propidium iodide, a well-known example, has limitations like low brightness, high toxicity, and poor bacterial differentiation. To address these challenges, we developed VersaDye, a modular probe designed for specific imaging of live plant cell walls and monitoring plasma membrane damage in plant cells, human cells, and certain bacteria. The design integrates impermeability principles and environment-dependent fluorophore scaffolds. VersaDye enables bright, wash-free labeling of plant cell walls and can stain various plant organs for constructing 3D tissue organization. Notably, it can selectively distinguish live Gram-positive from Gram-negative bacteria, a feature absent in other dyes. Its impermeability and targeting ability also allow it to probe membrane damage caused by physical, chemical, and biological stimuli. This study marks the first use of VersaDye in analyzing cell damage in live plants under salt stress. VersaDye offers a robust platform for wash-free, in vivo membrane damage monitoring and simultaneous cell wall labeling. Additionally, its design suggests adaptability for regulating permeability to meet specific diagnostic needs, such as identifying membrane-compromised cells in diseases or enabling high-throughput antibiotic screening targeting specific bacteria.
{"title":"Modular Design of Membrane-Impermeable Versatile Probe for Specific Imaging of Cell Walls and Real-Time Detection of Cell Membrane Damage","authors":"Zhengdong Han, Tian Li, Ziqing He, Engao Zhu, Zhaosheng Qian, Xia Liu, Hui Feng","doi":"10.1021/acs.analchem.5c01229","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c01229","url":null,"abstract":"The versatile fluorescent dyes are essential for specifically labeling plant cell walls in vivo, monitoring plasma membrane damage, and assessing cell viability. However, such dyes are rare and often discovered accidentally due to a lack of design principles. Propidium iodide, a well-known example, has limitations like low brightness, high toxicity, and poor bacterial differentiation. To address these challenges, we developed VersaDye, a modular probe designed for specific imaging of live plant cell walls and monitoring plasma membrane damage in plant cells, human cells, and certain bacteria. The design integrates impermeability principles and environment-dependent fluorophore scaffolds. VersaDye enables bright, wash-free labeling of plant cell walls and can stain various plant organs for constructing 3D tissue organization. Notably, it can selectively distinguish live Gram-positive from Gram-negative bacteria, a feature absent in other dyes. Its impermeability and targeting ability also allow it to probe membrane damage caused by physical, chemical, and biological stimuli. This study marks the first use of VersaDye in analyzing cell damage in live plants under salt stress. VersaDye offers a robust platform for wash-free, in vivo membrane damage monitoring and simultaneous cell wall labeling. Additionally, its design suggests adaptability for regulating permeability to meet specific diagnostic needs, such as identifying membrane-compromised cells in diseases or enabling high-throughput antibiotic screening targeting specific bacteria.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"16 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143766651","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}
Efficient ionization is essential for sensitive mass spectrometry (MS) analysis. Herein, a novel laser-assisted electrospray ionization (laser-assisted ESI) source was developed to efficiently ionize low-polar and thermal unstable compounds. By irradiating the electrospray nozzle with a simple and low-cost laser probe (450 nm, 500 mW), the sample droplets were stimulated with a laser beam as well as the high voltage of the electrospray, which significantly enhanced the ionization efficiency. In positive ionization mode, by further utilizing the reaction of Ag+ and tetrabromobisphenol A bis(allyl ether) (TBBPA-BAE), the established laser-assisted ESI strategy was able to efficiently ionize the low-polar and thermal unstable TBBPA-BAE. Specifically, a limit of detection (LOD) of 14 ng L–1, linear range of 0.1–10 μg L–1 (R2 > 0.99), and relative standard deviations (RSDs) of 2.5% (n = 7, intraday) and 6.2% (n = 3 per day for 5 days, interday) were achieved. In negative ionization mode, laser-assisted ESI also improved the detection sensitivity of tetrabromobisphenol A (TBBPA), achieving a LOD of 3.3 ng L–1, linear range of 0.01–10 μg L–1 (R2 > 0.99), and RSDs of 4.7% (n = 7, intraday) and 7.3% (n = 3 per day for 5 days, interday). Compared to extractive electrospray ionization mass spectrometry (EESI-MS) and ESI-MS, the LODs achieved with laser-assisted ESI-MS were 54 and 16 times lower for the detection of TBBPA-BAE and TBBPA, respectively. Notably, deep purification and preconcentration were not required to accurately detect TBBPA and TBBPA-BAE in river water and wastewater treatment plant effluent. The spiked recoveries were between 88.0% and 106.0%, demonstrating the high reliability and practicality of this method.
{"title":"Novel Laser-Assisted Electrospray Ionization Mass Spectrometry (Laser-Assisted ESI-MS): A Sensitive Method for Determining Tetrabromobisphenol A and Its Derivative","authors":"Haonan Liu, Xiaoxuan Han, Shuo Gao, Fada Shi, Yong Tian, Ligang Hu, Jianbo Shi, Guibin Jiang","doi":"10.1021/acs.analchem.5c00166","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c00166","url":null,"abstract":"Efficient ionization is essential for sensitive mass spectrometry (MS) analysis. Herein, a novel laser-assisted electrospray ionization (laser-assisted ESI) source was developed to efficiently ionize low-polar and thermal unstable compounds. By irradiating the electrospray nozzle with a simple and low-cost laser probe (450 nm, 500 mW), the sample droplets were stimulated with a laser beam as well as the high voltage of the electrospray, which significantly enhanced the ionization efficiency. In positive ionization mode, by further utilizing the reaction of Ag<sup>+</sup> and tetrabromobisphenol A bis(allyl ether) (TBBPA-BAE), the established laser-assisted ESI strategy was able to efficiently ionize the low-polar and thermal unstable TBBPA-BAE. Specifically, a limit of detection (LOD) of 14 ng L<sup>–1</sup>, linear range of 0.1–10 μg L<sup>–1</sup> (R<sup>2</sup> > 0.99), and relative standard deviations (RSDs) of 2.5% (n = 7, intraday) and 6.2% (n = 3 per day for 5 days, interday) were achieved. In negative ionization mode, laser-assisted ESI also improved the detection sensitivity of tetrabromobisphenol A (TBBPA), achieving a LOD of 3.3 ng L<sup>–1</sup>, linear range of 0.01–10 μg L<sup>–1</sup> (R<sup>2</sup> > 0.99), and RSDs of 4.7% (n = 7, intraday) and 7.3% (n = 3 per day for 5 days, interday). Compared to extractive electrospray ionization mass spectrometry (EESI-MS) and ESI-MS, the LODs achieved with laser-assisted ESI-MS were 54 and 16 times lower for the detection of TBBPA-BAE and TBBPA, respectively. Notably, deep purification and preconcentration were not required to accurately detect TBBPA and TBBPA-BAE in river water and wastewater treatment plant effluent. The spiked recoveries were between 88.0% and 106.0%, demonstrating the high reliability and practicality of this method.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"58 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143766652","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}
Mycotoxins, particularly deoxynivalenol (DON) and zearalenone (ZEN), are common food contaminants that frequently co-occur in grains, posing significant health risks. This study proposed a multiplexed detection platform for simultaneous quantification and imaging of three microRNAs (miRNAs) integrated with machine learning to evaluate the combined toxicity of DON and ZEN. Based on Exonuclease III-assisted signal amplification, highly sensitive fluorescent molecular beacon probes (MBs) targeting miR-21, miR-221, and miR-27a were developed, achieving remarkable detection limits of 0.18 pM, 0.22 pM, and 0.21 pM, respectively. The MBs were efficiently delivered into cells via liposome-mediated endocytosis, enabling simultaneous intracellular imaging of the three miRNAs. By integrating machine learning algorithms, including linear discriminant analysis and principal component analysis, with RGB values extracted from cellular fluorescence images, a robust analytical platform was established for classifying miRNA expression patterns induced by various DON/ZEN concentrations. A highest single agent model was subsequently constructed to evaluate the combined toxicity, revealing that ZEN exhibited antagonistic effects on DON at low doses but synergistic effects at high doses. This sensitive and multiplexed detection method demonstrates a strong correlation between miRNA expression profiles and DON/ZEN toxicity, providing an innovative analytical tool for multicomponent toxicity assessment.
{"title":"Machine Learning-Assisted Multiplexed Fluorescence-Labeled miRNAs Imaging Decoding for Combined Mycotoxins Toxicity Assessment","authors":"Lixin Kang, Xianfeng Lin, Jiaqi Feng, Mengxia Duan, Nuo Duan, Zhouping Wang, Shijia Wu","doi":"10.1021/acs.analchem.5c00404","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c00404","url":null,"abstract":"Mycotoxins, particularly deoxynivalenol (DON) and zearalenone (ZEN), are common food contaminants that frequently co-occur in grains, posing significant health risks. This study proposed a multiplexed detection platform for simultaneous quantification and imaging of three microRNAs (miRNAs) integrated with machine learning to evaluate the combined toxicity of DON and ZEN. Based on Exonuclease III-assisted signal amplification, highly sensitive fluorescent molecular beacon probes (MBs) targeting miR-21, miR-221, and miR-27a were developed, achieving remarkable detection limits of 0.18 pM, 0.22 pM, and 0.21 pM, respectively. The MBs were efficiently delivered into cells via liposome-mediated endocytosis, enabling simultaneous intracellular imaging of the three miRNAs. By integrating machine learning algorithms, including linear discriminant analysis and principal component analysis, with RGB values extracted from cellular fluorescence images, a robust analytical platform was established for classifying miRNA expression patterns induced by various DON/ZEN concentrations. A highest single agent model was subsequently constructed to evaluate the combined toxicity, revealing that ZEN exhibited antagonistic effects on DON at low doses but synergistic effects at high doses. This sensitive and multiplexed detection method demonstrates a strong correlation between miRNA expression profiles and DON/ZEN toxicity, providing an innovative analytical tool for multicomponent toxicity assessment.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"107 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143767025","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}
Pub Date : 2025-04-03DOI: 10.1021/acs.analchem.4c07055
Chengjie Huang, Tianbao Ye, Xiuyuan Wang, Ke Li, Yiyang Li, Lai Jiang, Xianting Ding
Mass cytometry (CyTOF) and imaging mass cytometry (IMC), as cutting-edge technologies in single-cell analysis, are capable of detecting more than 40 biomarkers simultaneously on a single cell. However, their sensitivity and multiparameter detection capabilities have been long constrained by the development of metal labeling materials. Meanwhile, as an imaging technique, IMC has suffered from a rather slow data acquisition rate. Here, we present a luminescent PCN-224-OH material that exhibits both fluorescent and mass dual-functionality and is enriched with Zr–OH–/H2O active sites. Without the additional need for complex postmodification or chemical coupling reactions, PCN-224-OH can be directly functionalized with antibodies/aptamers and poly(ethylene glycol) (PEG), resulting in the production of PCN-224-Ab-PEG or PCN-224-Apt-PEG probes. We demonstrated that PCN-224-Ab-PEG was compatible with commercial polymer-based probes but with superior sensitivity and specificity. Meanwhile, since PCN-224-Apt-PEG expressed both fluorescence and mass signals, we could adopt fluorescence signals for rapid tissue section scanning to swiftly identify the regions of interest (ROIs), and then adopt IMC for multiparameter imaging at the specific ROIs. The application of the PCN-224-Apt-PEG probe could significantly reduce the blind IMC scanning time by up to 90% and effectively compensate for IMC’s low resolution. This study not only broadens the application scope of luminescent metal–organic frameworks but also offers a potentially novel toolbox for single-cell multiparameter detection.
{"title":"Luminescent Metal–Organic Framework Probes with Metallic and Fluorescent Dual-Properties for Mass Cytometry and Imaging","authors":"Chengjie Huang, Tianbao Ye, Xiuyuan Wang, Ke Li, Yiyang Li, Lai Jiang, Xianting Ding","doi":"10.1021/acs.analchem.4c07055","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c07055","url":null,"abstract":"Mass cytometry (CyTOF) and imaging mass cytometry (IMC), as cutting-edge technologies in single-cell analysis, are capable of detecting more than 40 biomarkers simultaneously on a single cell. However, their sensitivity and multiparameter detection capabilities have been long constrained by the development of metal labeling materials. Meanwhile, as an imaging technique, IMC has suffered from a rather slow data acquisition rate. Here, we present a luminescent PCN-224-OH material that exhibits both fluorescent and mass dual-functionality and is enriched with Zr–OH<sup>–</sup>/H<sub>2</sub>O active sites. Without the additional need for complex postmodification or chemical coupling reactions, PCN-224-OH can be directly functionalized with antibodies/aptamers and poly(ethylene glycol) (PEG), resulting in the production of PCN-224-Ab-PEG or PCN-224-Apt-PEG probes. We demonstrated that PCN-224-Ab-PEG was compatible with commercial polymer-based probes but with superior sensitivity and specificity. Meanwhile, since PCN-224-Apt-PEG expressed both fluorescence and mass signals, we could adopt fluorescence signals for rapid tissue section scanning to swiftly identify the regions of interest (ROIs), and then adopt IMC for multiparameter imaging at the specific ROIs. The application of the PCN-224-Apt-PEG probe could significantly reduce the blind IMC scanning time by up to 90% and effectively compensate for IMC’s low resolution. This study not only broadens the application scope of luminescent metal–organic frameworks but also offers a potentially novel toolbox for single-cell multiparameter detection.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"216 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143766649","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}
Pub Date : 2025-04-03DOI: 10.1021/acs.analchem.4c07123
Haoni Yan, Yan Zhang, Yujie Shi, Jiahui Ding, Hengxing Su, Wenqiong Su, Yan Wang, Yanfei Mao, Tawfik A. Khattab, Salhah D. Al-Qahtani, Aynur Abdulla, Lai Jiang, Xianting Ding
Sepsis, a lethal organ dysfunction caused by a dysregulated host response to infection, is the leading cause of worldwide in-hospital mortality. However, the early diagnostic methods for sepsis are still urgent for guiding accurate antibiotic usage and improving the survival rate of the patients. Herein, we constructed a PD-L1 antibody affinity microfluidic (PAAM) chip for early sepsis diagnosis and severity assessment. The chip was used to capture PD-L1-expressing leukocytes from whole blood samples obtained from healthy control (HC) volunteers (n = 15) and sepsis patients on day 1 (D1) and day 7 (D7) (n = 20), and there was a statistically significant difference between HC and sepsis patients (p < 0.0001), and the AUC was 0.96. However, there was no significant difference in the number of cells captured on-chip between sepsis patients on D1 and D7 (p = 0.16). Therefore, we performed immunofluorescence staining of PD-L1, CD64, and CD123 on the chip. The results showed that the combination of PD-L1, CD64, and CD123 for sepsis diagnosis had an AUC of 0.98, and there was a significant difference in PD-L1+/CD64+/CD123+ leukocytes between sepsis patients on D1 and on D7 (p < 0.0001). In conclusion, we found that the combination of multiple biomarkers was more precise and dependable for sepsis diagnosis and severity assessment.
{"title":"Combining CD64 and CD123 Biomarkers for Sepsis Early Diagnosis and Severity Assessment via PD-L1 Antibody Affinity Microfluidic (PAAM) Chip in Trace Clinical Samples","authors":"Haoni Yan, Yan Zhang, Yujie Shi, Jiahui Ding, Hengxing Su, Wenqiong Su, Yan Wang, Yanfei Mao, Tawfik A. Khattab, Salhah D. Al-Qahtani, Aynur Abdulla, Lai Jiang, Xianting Ding","doi":"10.1021/acs.analchem.4c07123","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c07123","url":null,"abstract":"Sepsis, a lethal organ dysfunction caused by a dysregulated host response to infection, is the leading cause of worldwide in-hospital mortality. However, the early diagnostic methods for sepsis are still urgent for guiding accurate antibiotic usage and improving the survival rate of the patients. Herein, we constructed a PD-L1 antibody affinity microfluidic (PAAM) chip for early sepsis diagnosis and severity assessment. The chip was used to capture PD-L1-expressing leukocytes from whole blood samples obtained from healthy control (HC) volunteers (<i>n</i> = 15) and sepsis patients on day 1 (D1) and day 7 (D7) (<i>n</i> = 20), and there was a statistically significant difference between HC and sepsis patients (<i>p</i> < 0.0001), and the AUC was 0.96. However, there was no significant difference in the number of cells captured on-chip between sepsis patients on D1 and D7 (<i>p</i> = 0.16). Therefore, we performed immunofluorescence staining of PD-L1, CD64, and CD123 on the chip. The results showed that the combination of PD-L1, CD64, and CD123 for sepsis diagnosis had an AUC of 0.98, and there was a significant difference in PD-L1<sup>+</sup>/CD64<sup>+</sup>/CD123<sup>+</sup> leukocytes between sepsis patients on D1 and on D7 (<i>p</i> < 0.0001). In conclusion, we found that the combination of multiple biomarkers was more precise and dependable for sepsis diagnosis and severity assessment.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"34 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143766650","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}
Compound 6:2 chlorinated polyfluoroalkyl ether sulfonate (6:2 Cl-PFESA) is an emerging per- and polyfluoroalkyl substance (PFAS) with potential toxicity and health risks to biosystems and ecosystems. Here, we developed a metabolomics method based on single-cell mass spectrometry to investigate the hepatotoxicity and heterogeneous responses in zebrafish exposed to 6:2 Cl-PFESA. Zebrafish were exposed to an environmentally relevant concentration (200 ng/L) of 6:2 Cl-PFESA for 14 days. The livers were dissociated and prepared as cell suspensions and then introduced to high-throughput single-cell mass spectrometry for analysis of 6:2 Cl-PFESA and endogenous metabolites in individual primary liver cells. Significant sex-specific heterogeneity in 6:2 Cl-PFESA accumulation was observed (p < 0.05). Metabolomics analysis revealed perturbations in lipid metabolism, particularly affecting unsaturated fatty acids, ether lipids, and sphingolipids in zebrafish liver cells, indicating potential hepatotoxicity. Sex-dependent metabolic responses were evident: males showed notable changes in glucose and fatty acid metabolism, whereas females experienced pronounced disruptions in glycerophospholipid and amino acid pathways. ROC analysis identified sex-specific biomarkers, including FA(18:3) and FA(16:1) in males (AUC > 0.85), as well as proline and phosphatidylcholine in females (AUC > 0.90). These findings reflect metabolic dysregulation and highlight sex-specific responses. This study demonstrates the feasibility of single-cell metabolomics to elucidate the cellular mechanisms and metabolic responses of pollutant exposure, offering insights into precise and comprehensive toxicity assessments at the single-cell level.
{"title":"High-Throughput Single-Cell Mass Spectrometry Reveals Sex-Specific Metabolic Responses to 6:2 Chlorinated Polyfluoroalkyl Ether Sulfonate in Zebrafish Liver Cells","authors":"Chunfei Zhong, Haishen Zeng, Jiewei Deng, Quchang Li, Ziqing Li, Junqiu Zhai, Xinyan Li, Tiangang Luan","doi":"10.1021/acs.analchem.4c06345","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c06345","url":null,"abstract":"Compound 6:2 chlorinated polyfluoroalkyl ether sulfonate (6:2 Cl-PFESA) is an emerging per- and polyfluoroalkyl substance (PFAS) with potential toxicity and health risks to biosystems and ecosystems. Here, we developed a metabolomics method based on single-cell mass spectrometry to investigate the hepatotoxicity and heterogeneous responses in zebrafish exposed to 6:2 Cl-PFESA. Zebrafish were exposed to an environmentally relevant concentration (200 ng/L) of 6:2 Cl-PFESA for 14 days. The livers were dissociated and prepared as cell suspensions and then introduced to high-throughput single-cell mass spectrometry for analysis of 6:2 Cl-PFESA and endogenous metabolites in individual primary liver cells. Significant sex-specific heterogeneity in 6:2 Cl-PFESA accumulation was observed (<i>p</i> < 0.05). Metabolomics analysis revealed perturbations in lipid metabolism, particularly affecting unsaturated fatty acids, ether lipids, and sphingolipids in zebrafish liver cells, indicating potential hepatotoxicity. Sex-dependent metabolic responses were evident: males showed notable changes in glucose and fatty acid metabolism, whereas females experienced pronounced disruptions in glycerophospholipid and amino acid pathways. ROC analysis identified sex-specific biomarkers, including FA(18:3) and FA(16:1) in males (AUC > 0.85), as well as proline and phosphatidylcholine in females (AUC > 0.90). These findings reflect metabolic dysregulation and highlight sex-specific responses. This study demonstrates the feasibility of single-cell metabolomics to elucidate the cellular mechanisms and metabolic responses of pollutant exposure, offering insights into precise and comprehensive toxicity assessments at the single-cell level.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"32 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143766647","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}
Pub Date : 2025-04-02DOI: 10.1021/acs.analchem.5c00373
Lingling Luo, Yuying Zhou, Yaqin Chai, Ruo Yuan, Hongyan Liu
The minor changes of miRNA levels due to various diseases and cancers bring great challenges for early diagnosis. Here we propose a “signal on-off-super on” PEC biosensor based on a homogeneous multicycle cascaded DNA circuit and a SnSe/CdS photoanode for sensitive detection of biomarker miRNA-222. Specifically, a Z-type SnSe/CdS heterojunction with greatly enhanced photoanodic performance was developed to provide the initial “on” signal. The target miRNA-222 was converted to a dendritic DNA structure through a cascade DNA circuit. The PEC signal can be switched off by the dendritic DNA structure and further switched super on by the loading of photosensitizer manganese porphyrin (MnPP). It is worth noting that the homogeneous multicycle cascaded DNA circuit not only improved the reaction kinetics but also avoided the leakage of signal. Compared with the traditional “signal-on” or “signal-off” readout, this “signal on-off-super on” strategy avoids the false response and background, thereby enhancing the sensitivity and accuracy of the PEC biosensor. The detection limit of the constructed PEC sensor is 0.3 fM in the linear range from 1 fM to 10 nM. The PEC biosensor with outstanding reproducibility, stability, and sensitivity provides a promising platform for biomarker detection and early disease diagnosis.
{"title":"Homogeneous Multicycle Cascaded DNA Circuit for Sensitive “Signal On-Off-Super On” PEC Biosensing","authors":"Lingling Luo, Yuying Zhou, Yaqin Chai, Ruo Yuan, Hongyan Liu","doi":"10.1021/acs.analchem.5c00373","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c00373","url":null,"abstract":"The minor changes of miRNA levels due to various diseases and cancers bring great challenges for early diagnosis. Here we propose a “signal on-off-super on” PEC biosensor based on a homogeneous multicycle cascaded DNA circuit and a SnSe/CdS photoanode for sensitive detection of biomarker miRNA-222. Specifically, a Z-type SnSe/CdS heterojunction with greatly enhanced photoanodic performance was developed to provide the initial “on” signal. The target miRNA-222 was converted to a dendritic DNA structure through a cascade DNA circuit. The PEC signal can be switched off by the dendritic DNA structure and further switched super on by the loading of photosensitizer manganese porphyrin (MnPP). It is worth noting that the homogeneous multicycle cascaded DNA circuit not only improved the reaction kinetics but also avoided the leakage of signal. Compared with the traditional “signal-on” or “signal-off” readout, this “signal on-off-super on” strategy avoids the false response and background, thereby enhancing the sensitivity and accuracy of the PEC biosensor. The detection limit of the constructed PEC sensor is 0.3 fM in the linear range from 1 fM to 10 nM. The PEC biosensor with outstanding reproducibility, stability, and sensitivity provides a promising platform for biomarker detection and early disease diagnosis.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"128 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143766655","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}
Pub Date : 2025-04-02DOI: 10.1021/acs.analchem.4c05228
Phuong T. Do, Bailey Richardson, Samuel C. Brydon, Therese M. Fulloon, Stephen J. Blanksby, Hendrik Frisch, Berwyck L. J. Poad
[2 + 2] Photocycloaddition reactions are powerful tools for synthetic chemistry. However, analysis of the head-to-head or head-to-tail conformation of the resulting cycloadducts is often challenging by conventional spectroscopic methods. Herein, we report the analysis of coumarin and styrylpyrene cycloadducts by cyclic ion-mobility tandem mass spectrometry (cIM-MS/MS) to characterize the regioisomeric products of this important class of photoresponsive groups. Photodissociation (PD) and collision-induced dissociation (CID) of the cycloadduct ions in the gas phase gave similar products to photodissociation in solution with regiospecific fragmentation of the core cyclobutane ring. The styrylpyrene cycloadduct ion was observed to be more stable than the coumarin analog under CID conditions, reflecting the impact of different substituents on the stability of the cyclobutane ring. Exploiting the difference in cyclobutane fragmentation for head-to-head and head-to-tail styrylpyrene cycloadduct isomers, ion mobility enabled CID-MS/MS was applied successfully to differentiate and identify these isomers. The developed method proved to be robust even to complex molecular structures and enabled the identification and separation of photocycloadducts resulting from styrylpyrene terminated peptides, providing access to a rapid analysis of challenging cycloadduct isomers.
{"title":"Mass Spectrometry Directed Structural Elucidation of Isomeric [2 + 2] Photocycloadducts","authors":"Phuong T. Do, Bailey Richardson, Samuel C. Brydon, Therese M. Fulloon, Stephen J. Blanksby, Hendrik Frisch, Berwyck L. J. Poad","doi":"10.1021/acs.analchem.4c05228","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c05228","url":null,"abstract":"[2 + 2] Photocycloaddition reactions are powerful tools for synthetic chemistry. However, analysis of the head-to-head or head-to-tail conformation of the resulting cycloadducts is often challenging by conventional spectroscopic methods. Herein, we report the analysis of coumarin and styrylpyrene cycloadducts by cyclic ion-mobility tandem mass spectrometry (cIM-MS/MS) to characterize the regioisomeric products of this important class of photoresponsive groups. Photodissociation (PD) and collision-induced dissociation (CID) of the cycloadduct ions in the gas phase gave similar products to photodissociation in solution with regiospecific fragmentation of the core cyclobutane ring. The styrylpyrene cycloadduct ion was observed to be more stable than the coumarin analog under CID conditions, reflecting the impact of different substituents on the stability of the cyclobutane ring. Exploiting the difference in cyclobutane fragmentation for head-to-head and head-to-tail styrylpyrene cycloadduct isomers, ion mobility enabled CID-MS/MS was applied successfully to differentiate and identify these isomers. The developed method proved to be robust even to complex molecular structures and enabled the identification and separation of photocycloadducts resulting from styrylpyrene terminated peptides, providing access to a rapid analysis of challenging cycloadduct isomers.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"89 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143758272","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}
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