Pub Date : 2024-07-04DOI: 10.1021/acs.analchem.4c01479
Giulia Bonaccorso, Lapo Renai, Leonardo Checchini, Massimo Del Bubba
A new self-assembled apparatus for the extraction of solid samples was designed and implemented to perform a recirculated pressurized hot water extraction (R-PHWE) directly coupled to liquid chromatography-tandem mass spectrometry. To investigate the potential of this new extraction apparatus, 34 target pharmaceutical compounds were analyzed in loam, silt-loam, and silty-clay-loam soils. The target analytes were characterized by heterogeneous physicochemical properties (e.g., -1.60 ≤ log D ≤ 5.91 at pH = 7.2, i.e., at the mean pH values of the three soils). Design of experiments (DoE) was used to identify the best extraction conditions for the target analytes by studying temperature, pressure, and number of extraction cycles. The results of DoE optimization pointed out the significant influence of the number of cycles on recovery. The application of DoE set point to the three reference soils provided recoveries ≥60% for 21-25 out the 34 target analytes, depending on soil. Good recovery precision (<25%) and moderate suppressive matrix effect (≤40%) were found for most target analytes, regardless of the soil considered. The optimized R-PHWE procedure evidenced statistically higher recoveries for 16 out of 34 target analytes when compared to conventional off-line dynamic PHWE.
{"title":"A Novel Apparatus for the Fully Automated Extraction and Online Liquid Chromatographic Analysis of Solid Environmental Samples: Application to the Pressurized Hot Water Extraction of Pharmaceuticals in Soil.","authors":"Giulia Bonaccorso, Lapo Renai, Leonardo Checchini, Massimo Del Bubba","doi":"10.1021/acs.analchem.4c01479","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c01479","url":null,"abstract":"<p><p>A new self-assembled apparatus for the extraction of solid samples was designed and implemented to perform a recirculated pressurized hot water extraction (R-PHWE) directly coupled to liquid chromatography-tandem mass spectrometry. To investigate the potential of this new extraction apparatus, 34 target pharmaceutical compounds were analyzed in loam, silt-loam, and silty-clay-loam soils. The target analytes were characterized by heterogeneous physicochemical properties (e.g., -1.60 ≤ log <i>D</i> ≤ 5.91 at pH = 7.2, i.e., at the mean pH values of the three soils). Design of experiments (DoE) was used to identify the best extraction conditions for the target analytes by studying temperature, pressure, and number of extraction cycles. The results of DoE optimization pointed out the significant influence of the number of cycles on recovery. The application of DoE set point to the three reference soils provided recoveries ≥60% for 21-25 out the 34 target analytes, depending on soil. Good recovery precision (<25%) and moderate suppressive matrix effect (≤40%) were found for most target analytes, regardless of the soil considered. The optimized R-PHWE procedure evidenced statistically higher recoveries for 16 out of 34 target analytes when compared to conventional off-line dynamic PHWE.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":null,"pages":null},"PeriodicalIF":6.7,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141532851","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 : 2024-07-04DOI: 10.1021/acs.analchem.4c01912
Zhichao Yu, Juan Tang, Man Xu, Di Wu, Yuan Gao, Yongyi Zeng, Xiaolong Liu, Dianping Tang
In this work, we reported a cholesterol oxidase (Chox)-loaded platinum (Pt) nanozyme with the collaborative cascade nanoreactor for the construction of nanozyme-enzyme-linked immunosorbent assay (N-ELSA) models to realize high-throughput rapid evaluation of cancer markers. Considering the high specific surface area and manipulable surface sites, ZIF-8 was used as a substrate for natural enzyme and nanozyme loading. The constructed ZIF-8-Pt nanozyme platform exhibited efficient enzyme-like catalytic efficiency with a standard corrected activity of 60.59 U mg-1, which was 12 times higher than that of the ZIF-8 precursor, and highly efficient photothermal conversion efficiency (∼35.49%). In N-ELISA testing, developed multienzyme photothermal probes were immobilized in microplates based on antigen-antibody-specific reactions. Cholesterol was reacted in a cascade to reactive oxygen radicals, which attacked 3,3',5,5'-tetramethylbenzidine, causing it to oxidize and color change, thus exhibiting highly enhanced efficient photothermal properties. Systematic temperature evaluations were performed by a hand-held microelectromechanical system thermal imager under the excitation of an 808 nm surface light source to determine the cancer antigen 15-3 (CA15-3) profiles in the samples. Encouragingly, the temperature signal from the microwells increased with increasing CA15-3, with a linear range of 2 mU mL-1 to 100 U mL-1, considering it to be the sensor with the widest working range for visualization and portability available. This work provides new horizons for the development of efficient multienzyme portable colorimetric-photothermal platforms to help advance the community-based process of early cancer detection.
在这项工作中,我们报道了一种负载胆固醇氧化酶(Chox)的铂(Pt)纳米酶与协作级联纳米反应器用于构建纳米酶-酶联免疫吸附测定(N-ELSA)模型,以实现癌症标志物的高通量快速评估。考虑到高比表面积和可操作的表面位点,ZIF-8 被用作天然酶和纳米酶负载的底物。所构建的 ZIF-8-Pt 纳米酶平台具有高效的酶样催化效率,其标准校正活性为 60.59 U mg-1,是 ZIF-8 前体的 12 倍,并具有高效的光热转换效率(∼35.49%)。在 N-ELISA 测试中,开发的多酶光热探针被固定在基于抗原-抗体特异性反应的微孔板中。胆固醇与活性氧自由基发生级联反应,攻击 3,3',5,5'-四甲基联苯胺,使其氧化变色,从而表现出高度增强的高效光热特性。在 808 纳米表面光源的激发下,手持式微机电系统热成像仪对样品进行了系统的温度评估,以确定样品中癌症抗原 15-3 (CA15-3) 的分布情况。令人鼓舞的是,微孔的温度信号随着 CA15-3 的增加而增加,线性范围从 2 mU mL-1 到 100 U mL-1,是目前可视化和便携性工作范围最广的传感器。这项工作为开发高效的多酶便携式比色-光热平台开辟了新天地,有助于推进基于社区的早期癌症检测进程。
{"title":"Multi-Enzyme Cascade Nanoreactors for High-Throughput Immunoassay: Transitioning Concept in Lab to Application in Community.","authors":"Zhichao Yu, Juan Tang, Man Xu, Di Wu, Yuan Gao, Yongyi Zeng, Xiaolong Liu, Dianping Tang","doi":"10.1021/acs.analchem.4c01912","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c01912","url":null,"abstract":"<p><p>In this work, we reported a cholesterol oxidase (Chox)-loaded platinum (Pt) nanozyme with the collaborative cascade nanoreactor for the construction of nanozyme-enzyme-linked immunosorbent assay (N-ELSA) models to realize high-throughput rapid evaluation of cancer markers. Considering the high specific surface area and manipulable surface sites, ZIF-8 was used as a substrate for natural enzyme and nanozyme loading. The constructed ZIF-8-Pt nanozyme platform exhibited efficient enzyme-like catalytic efficiency with a standard corrected activity of 60.59 U mg<sup>-1</sup>, which was 12 times higher than that of the ZIF-8 precursor, and highly efficient photothermal conversion efficiency (∼35.49%). In N-ELISA testing, developed multienzyme photothermal probes were immobilized in microplates based on antigen-antibody-specific reactions. Cholesterol was reacted in a cascade to reactive oxygen radicals, which attacked 3,3',5,5'-tetramethylbenzidine, causing it to oxidize and color change, thus exhibiting highly enhanced efficient photothermal properties. Systematic temperature evaluations were performed by a hand-held microelectromechanical system thermal imager under the excitation of an 808 nm surface light source to determine the cancer antigen 15-3 (CA15-3) profiles in the samples. Encouragingly, the temperature signal from the microwells increased with increasing CA15-3, with a linear range of 2 mU mL<sup>-1</sup> to 100 U mL<sup>-1</sup>, considering it to be the sensor with the widest working range for visualization and portability available. This work provides new horizons for the development of efficient multienzyme portable colorimetric-photothermal platforms to help advance the community-based process of early cancer detection.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":null,"pages":null},"PeriodicalIF":6.7,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141496309","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 : 2024-07-03DOI: 10.1021/acs.analchem.4c01002
Amina Souihi, Anneli Kruve
Nontargeted LC/ESI/HRMS aims to detect and identify organic compounds present in the environment without prior knowledge; however, in practice no LC/ESI/HRMS method is capable of detecting all chemicals, and the scope depends on the instrumental conditions. Different experimental conditions, instruments, and methods used for sample preparation and nontargeted LC/ESI/HRMS as well as different workflows for data processing may lead to challenges in communicating the results and sharing data between laboratories as well as reduced reproducibility. One of the reasons is that only a fraction of method performance characteristics can be determined for a nontargeted analysis method due to the lack of prior information and analytical standards of the chemicals present in the sample. The limit of detection (LoD) is one of the most important performance characteristics in target analysis and directly describes the detectability of a chemical. Recently, the identification and quantification in nontargeted LC/ESI/HRMS (e.g., via predicting ionization efficiency, risk scores, and retention times) have significantly improved due to employing machine learning. In this work, we hypothesize that the predicted ionization efficiency could be used to estimate LoD and thereby enable evaluating the suitability of the LC/ESI/HRMS nontargeted method for the detection of suspected chemicals even if analytical standards are lacking. For this, 221 representative compounds were selected from the NORMAN SusDat list (S0), and LoD values were determined by using 4 complementary approaches. The LoD values were correlated to ionization efficiency values predicted with previously trained random forest regression. A robust regression was then used to estimate LoD values of unknown features detected in the nontargeted screening of wastewater samples. These estimated LoD values were used for prioritization of the unknown features. Furthermore, we present LoD values for the NORMAN SusDat list with a reversed-phase C18 LC method.
{"title":"Estimating LoD-s Based on the Ionization Efficiency Values for the Reporting and Harmonization of Amenable Chemical Space in Nontargeted Screening LC/ESI/HRMS.","authors":"Amina Souihi, Anneli Kruve","doi":"10.1021/acs.analchem.4c01002","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c01002","url":null,"abstract":"<p><p>Nontargeted LC/ESI/HRMS aims to detect and identify organic compounds present in the environment without prior knowledge; however, in practice no LC/ESI/HRMS method is capable of detecting all chemicals, and the scope depends on the instrumental conditions. Different experimental conditions, instruments, and methods used for sample preparation and nontargeted LC/ESI/HRMS as well as different workflows for data processing may lead to challenges in communicating the results and sharing data between laboratories as well as reduced reproducibility. One of the reasons is that only a fraction of method performance characteristics can be determined for a nontargeted analysis method due to the lack of prior information and analytical standards of the chemicals present in the sample. The limit of detection (LoD) is one of the most important performance characteristics in target analysis and directly describes the detectability of a chemical. Recently, the identification and quantification in nontargeted LC/ESI/HRMS (e.g., via predicting ionization efficiency, risk scores, and retention times) have significantly improved due to employing machine learning. In this work, we hypothesize that the predicted ionization efficiency could be used to estimate LoD and thereby enable evaluating the suitability of the LC/ESI/HRMS nontargeted method for the detection of suspected chemicals even if analytical standards are lacking. For this, 221 representative compounds were selected from the NORMAN SusDat list (S0), and LoD values were determined by using 4 complementary approaches. The LoD values were correlated to ionization efficiency values predicted with previously trained random forest regression. A robust regression was then used to estimate LoD values of unknown features detected in the nontargeted screening of wastewater samples. These estimated LoD values were used for prioritization of the unknown features. Furthermore, we present LoD values for the NORMAN SusDat list with a reversed-phase C<sub>18</sub> LC method.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":null,"pages":null},"PeriodicalIF":6.7,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141496307","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 : 2024-07-03DOI: 10.1021/acs.analchem.4c01170
Jan Konrad Wied, Benjamin Mockenhaupt, Ulrich Schürmann, Lorenz Kienle, Sebastian Mangelsen, Janin Glänzer, Vinicius Ribeiro Celinski, Malte Behrens, Jörn Schmedt Auf der Günne
Nanoscale zinc-oxide doped with aluminum ZnO:Al is studied by different techniques targeting surface changes induced by the conditions at which ZnO:Al is used as support material in the catalysis of methanol. While it is well established that a variety of 1H and 27Al resonances can be found by solid-state NMR for this material, it was not clear yet which signals are related to species located close to the surface of the material and which to species located in the bulk. To this end, a method is suggested that makes use of a paramagnetically impregnated material to suppress NMR signals close to the particle surface in the blind sphere around the paramagnetic metal atoms. It is shown that it is important to use conditions that guarantee a stable reference system relative to which it can be established whether the coating procedure is conserving the original structure or not. This method, called paramagnetically assisted surface peak assignment, helped to assign the 1H and 27Al NMR peaks to the bulk and the surface layer defined by the blind sphere of the paramagnetic atoms. The assignment results are further corroborated by the results from heteronuclear 27Al{1H} dipolar dephasing experiments, which indicate that the hydrogen atoms are preferentially located in the surface layer and not in the particle core.
{"title":"Method for Surface Characterization Using Solid-State Nuclear Magnetic Resonance Spectroscopy Demonstrated on Nanocrystalline ZnO:Al.","authors":"Jan Konrad Wied, Benjamin Mockenhaupt, Ulrich Schürmann, Lorenz Kienle, Sebastian Mangelsen, Janin Glänzer, Vinicius Ribeiro Celinski, Malte Behrens, Jörn Schmedt Auf der Günne","doi":"10.1021/acs.analchem.4c01170","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c01170","url":null,"abstract":"<p><p>Nanoscale zinc-oxide doped with aluminum ZnO:Al is studied by different techniques targeting surface changes induced by the conditions at which ZnO:Al is used as support material in the catalysis of methanol. While it is well established that a variety of <sup>1</sup>H and <sup>27</sup>Al resonances can be found by solid-state NMR for this material, it was not clear yet which signals are related to species located close to the surface of the material and which to species located in the bulk. To this end, a method is suggested that makes use of a paramagnetically impregnated material to suppress NMR signals close to the particle surface in the blind sphere around the paramagnetic metal atoms. It is shown that it is important to use conditions that guarantee a stable reference system relative to which it can be established whether the coating procedure is conserving the original structure or not. This method, called paramagnetically assisted surface peak assignment, helped to assign the <sup>1</sup>H and <sup>27</sup>Al NMR peaks to the bulk and the surface layer defined by the blind sphere of the paramagnetic atoms. The assignment results are further corroborated by the results from heteronuclear <sup>27</sup>Al{<sup>1</sup>H} dipolar dephasing experiments, which indicate that the hydrogen atoms are preferentially located in the surface layer and not in the particle core.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":null,"pages":null},"PeriodicalIF":6.7,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141489931","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}
Microalgae metabolite analysis is fundamental for the rational design of metabolic engineering strategies for the biosynthesis of high-value products. Mass spectrometry (MS) has been utilized for single-cell microalgae analysis. However, limitations in the detection throughput and polarities of detectable substances make it difficult to realize high-throughput screening of high-performance microalgae. Herein, a plasma-assisted label-free mass cytometry, named as PACyESI-MS, was proposed combining the advantages of orthogonal hybrid ionization and high-throughput MS analysis, which realized rapid metabolite detection of single microalgae. The cell detection throughput of PACyESI-MS was up to 52 cells/min. Dozens of the critical primary and secondary metabolites within single microalgae were detected simultaneously, including pigments, lipids, and energy metabolites. Furthermore, metabolite changes of Chlamydomonas reinhardtii and Haematococcus pluvialis under nitrogen deficiency stress were studied. Discrimination of Chlamydomonas under different nutrient conditions was realized using single-cell metabolite profiles obtained by PACyESI-MS. The relationships between the accumulation of bioactive astaxanthin and changes in functional primary metabolites of Haematococcus were investigated. It was demonstrated that PACyESI-MS can detect the flexible change of metabolites in single microalgae cells under different nutritional conditions and during the synthesis of high-value products, which is expected to become an important tool for the design of metabolic engineering-based high-performance microalgae factories.
{"title":"High-Throughput Metabolite Analysis of Unicellular Microalgae by Orthogonal Hybrid Ionization Label-Free Mass Cytometry.","authors":"Huan Yao, Jinlei Yang, Zhengmao Wang, Xingyu Pan, Junmin Pan, Hongmei Li, Sichun Zhang","doi":"10.1021/acs.analchem.4c01541","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c01541","url":null,"abstract":"<p><p>Microalgae metabolite analysis is fundamental for the rational design of metabolic engineering strategies for the biosynthesis of high-value products. Mass spectrometry (MS) has been utilized for single-cell microalgae analysis. However, limitations in the detection throughput and polarities of detectable substances make it difficult to realize high-throughput screening of high-performance microalgae. Herein, a plasma-assisted label-free mass cytometry, named as PACyESI-MS, was proposed combining the advantages of orthogonal hybrid ionization and high-throughput MS analysis, which realized rapid metabolite detection of single microalgae. The cell detection throughput of PACyESI-MS was up to 52 cells/min. Dozens of the critical primary and secondary metabolites within single microalgae were detected simultaneously, including pigments, lipids, and energy metabolites. Furthermore, metabolite changes of <i>Chlamydomonas reinhardtii</i> and <i>Haematococcus pluvialis</i> under nitrogen deficiency stress were studied. Discrimination of Chlamydomonas under different nutrient conditions was realized using single-cell metabolite profiles obtained by PACyESI-MS. The relationships between the accumulation of bioactive astaxanthin and changes in functional primary metabolites of Haematococcus were investigated. It was demonstrated that PACyESI-MS can detect the flexible change of metabolites in single microalgae cells under different nutritional conditions and during the synthesis of high-value products, which is expected to become an important tool for the design of metabolic engineering-based high-performance microalgae factories.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":null,"pages":null},"PeriodicalIF":6.7,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141496308","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 : 2024-07-03DOI: 10.1021/acs.analchem.4c01671
Yoshiyuki Tsuyama, Kazuma Mawatari
Characterization and quantification of plasmonic nanoparticles at the single particle level have become increasingly important with the advancements in nanotechnology and their application to various biological analyses including diagnostics, photothermal therapy, and immunoassays. While various nanoparticle detection methodologies have been developed and widely used, simultaneous measurement of light absorption and scattering from individual plasmonic nanoparticles in flow is still challenging. Herein, we describe a novel nanofluidic detection platform that enables simultaneous measurement of absorption and scattering signals from individual nanoparticles within a nanochannel. Our detection platform utilized optical diffraction phenomena by a single nanochannel as both a readout signal for photothermal detection and a reference light for interferometric scattering detection. Through the elucidation of the frequency effect on the detection performance and optimization of experimental conditions, we achieved the classification of gold and silver nanoparticles with a diameter of 20–60 nm at an average accuracy score of 82.6 ± 2.1% by measured data sets of absorption and scattering signals. Furthermore, we demonstrated the concentration determination of plasmonic nanoparticle mixtures using a trained Support vector machine (SVM) classifier. Our simple yet sensitive nanofluidic detection platform will be a valuable tool for the analysis of nanoparticles and their applications to chemical and biological assays.
{"title":"Nanofluidic Detection Platform for Simultaneous Light Absorption and Scattering Measurement of Individual Nanoparticles in Flow","authors":"Yoshiyuki Tsuyama, Kazuma Mawatari","doi":"10.1021/acs.analchem.4c01671","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c01671","url":null,"abstract":"Characterization and quantification of plasmonic nanoparticles at the single particle level have become increasingly important with the advancements in nanotechnology and their application to various biological analyses including diagnostics, photothermal therapy, and immunoassays. While various nanoparticle detection methodologies have been developed and widely used, simultaneous measurement of light absorption and scattering from individual plasmonic nanoparticles in flow is still challenging. Herein, we describe a novel nanofluidic detection platform that enables simultaneous measurement of absorption and scattering signals from individual nanoparticles within a nanochannel. Our detection platform utilized optical diffraction phenomena by a single nanochannel as both a readout signal for photothermal detection and a reference light for interferometric scattering detection. Through the elucidation of the frequency effect on the detection performance and optimization of experimental conditions, we achieved the classification of gold and silver nanoparticles with a diameter of 20–60 nm at an average accuracy score of 82.6 ± 2.1% by measured data sets of absorption and scattering signals. Furthermore, we demonstrated the concentration determination of plasmonic nanoparticle mixtures using a trained Support vector machine (SVM) classifier. Our simple yet sensitive nanofluidic detection platform will be a valuable tool for the analysis of nanoparticles and their applications to chemical and biological assays.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":null,"pages":null},"PeriodicalIF":7.4,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141495956","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}
It is well-known that the bacterial microenvironment imposes restrictions on the growth and behavior of bacteria. The localized monitoring of microenvironmental factors is appreciated when consulting bacterial adaptation and behavior in the presence of chemical or mechanical stimuli. Herein, we developed a novel liquid crystal (LC) biosensor in a microsphere configuration for real-time 3D monitoring of the bacteria microenvironment, which was implemented by a microfluidic chip. As a proof of concept, a LC gel (LC-Gel) microsphere biosensor was prepared and employed in the localized pH changes of bacteria by observing the configuration change of LC under polarized optical microscopy. Briefly, the microsphere biosensor was constructed in core-shell configuration, wherein the core contained LCE7 (a nematic LC) doped with 4-pentylbiphenyl-4'-carboxylic acid (PBA), and the shell encapsulated the bacteria. The protonation of carboxyl functional groups of the PBA induced a change in charge density on the surface of LCE7 and the orientation of E7 molecules, resulting in the transitions of the LC nucleus from axial to bipolar. The developed LC-Gel microspheres pH sensor exhibited its dominant performance on localized pH real-time sensing with a resolution of 0.1. An intriguing observation from the prepared pH biosensor was that the diverse bacteria impelled distinct acidifying or alkalizing effects. Overall, the facile LC-Gel microsphere biosensor not only provides a versatile tool for label-free, localized pH monitoring but also opens avenues for investigating the effects of chemical and mechanical stimuli on cellular metabolism within bacterial microenvironments.
{"title":"Real-Time pH Sensor in Bacterial Microenvironments Using Liquid Crystal Core-Shell Microspheres.","authors":"Yaoshuang Xie, Yuxuan Li, Haifeng Lin, Xiaorui Wang, Wenjun Liao, Zeyang Liu, Ling Lin","doi":"10.1021/acs.analchem.4c02040","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c02040","url":null,"abstract":"<p><p>It is well-known that the bacterial microenvironment imposes restrictions on the growth and behavior of bacteria. The localized monitoring of microenvironmental factors is appreciated when consulting bacterial adaptation and behavior in the presence of chemical or mechanical stimuli. Herein, we developed a novel liquid crystal (LC) biosensor in a microsphere configuration for real-time 3D monitoring of the bacteria microenvironment, which was implemented by a microfluidic chip. As a proof of concept, a LC gel (LC-Gel) microsphere biosensor was prepared and employed in the localized pH changes of bacteria by observing the configuration change of LC under polarized optical microscopy. Briefly, the microsphere biosensor was constructed in core-shell configuration, wherein the core contained LCE7 (a nematic LC) doped with 4-pentylbiphenyl-4'-carboxylic acid (PBA), and the shell encapsulated the bacteria. The protonation of carboxyl functional groups of the PBA induced a change in charge density on the surface of LCE7 and the orientation of E7 molecules, resulting in the transitions of the LC nucleus from axial to bipolar. The developed LC-Gel microspheres pH sensor exhibited its dominant performance on localized pH real-time sensing with a resolution of 0.1. An intriguing observation from the prepared pH biosensor was that the diverse bacteria impelled distinct acidifying or alkalizing effects. Overall, the facile LC-Gel microsphere biosensor not only provides a versatile tool for label-free, localized pH monitoring but also opens avenues for investigating the effects of chemical and mechanical stimuli on cellular metabolism within bacterial microenvironments.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":null,"pages":null},"PeriodicalIF":6.7,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141489868","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 : 2024-07-03DOI: 10.1021/acs.analchem.4c01564
Deben N Shoup, Sanjun Fan, Mario Zapata-Herrera, Hannah C Schorr, Javier Aizpurua, Zachary D Schultz
Strongly confined electric fields resulting from nanogaps within nanoparticle aggregates give rise to significant enhancement of surface-enhanced Raman scattering (SERS). Nanometer differences in gap sizes lead to drastically different confined field strengths; so much attention has been focused on the development and understanding of nanostructures with controlled gap sizes. In this work, we report a novel petal gap-enhanced Raman tag (GERT) consisting of a bipyramid core and a nitrothiophenol (NTP) spacer to support the growth of hundreds of small petals and compare its SERS emission and localization to a traditional bipyramid aggregate. To do this, we use super resolution spectral SERS imaging that simultaneously captures the SERS images and spectra while varying the incident laser polarization. Intensity fluctuations inherent of SERS enabled super resolution algorithms to be applied, which revealed subdiffraction limited differences in the localization with respect to polarization direction for both particles. Interestingly, however, only the traditional bipyramid aggregates experienced a strong polarization dependence in their SERS intensity and in the plasmon-induced conversion of NTP to dimercaptoazobenzene (DMAB), which was localized with nanometer precision to regions of intense electromagnetic fields. The lack of polarization dependence (validated through electromagnetic simulations) and surface reactions from the bipyramid-GERTs suggests that the emissions arising from the bipyramid-GERTs are less influenced by confined fields.
{"title":"Comparison of Gap-Enhanced Raman Tags and Nanoparticle Aggregates with Polarization Dependent Super-Resolution Spectral SERS Imaging.","authors":"Deben N Shoup, Sanjun Fan, Mario Zapata-Herrera, Hannah C Schorr, Javier Aizpurua, Zachary D Schultz","doi":"10.1021/acs.analchem.4c01564","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c01564","url":null,"abstract":"<p><p>Strongly confined electric fields resulting from nanogaps within nanoparticle aggregates give rise to significant enhancement of surface-enhanced Raman scattering (SERS). Nanometer differences in gap sizes lead to drastically different confined field strengths; so much attention has been focused on the development and understanding of nanostructures with controlled gap sizes. In this work, we report a novel petal gap-enhanced Raman tag (GERT) consisting of a bipyramid core and a nitrothiophenol (NTP) spacer to support the growth of hundreds of small petals and compare its SERS emission and localization to a traditional bipyramid aggregate. To do this, we use super resolution spectral SERS imaging that simultaneously captures the SERS images and spectra while varying the incident laser polarization. Intensity fluctuations inherent of SERS enabled super resolution algorithms to be applied, which revealed subdiffraction limited differences in the localization with respect to polarization direction for both particles. Interestingly, however, only the traditional bipyramid aggregates experienced a strong polarization dependence in their SERS intensity and in the plasmon-induced conversion of NTP to dimercaptoazobenzene (DMAB), which was localized with nanometer precision to regions of intense electromagnetic fields. The lack of polarization dependence (validated through electromagnetic simulations) and surface reactions from the bipyramid-GERTs suggests that the emissions arising from the bipyramid-GERTs are less influenced by confined fields.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":null,"pages":null},"PeriodicalIF":6.7,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141489924","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 : 2024-07-03DOI: 10.1021/acs.analchem.4c02251
Kenta Iitani, Mika Suzuki, Kenta Ichikawa, Koji Toma, Takahiro Arakawa, Kohji Mitsubayashi
Human-borne acetone is a potent marker of lipid metabolism. Here, an enzyme immobilization method for secondary alcohol dehydrogenase (S-ADH), which is suitable for highly sensitive and selective biosensing of acetone, was developed, and then its applicability was demonstrated for spatiotemporal imaging of concentration distribution. After various investigations, S-ADH-immobilized meshes could be prepared with less than 5% variation by cross-linking S-ADH with glutaraldehyde on a cotton mesh at 40 °C for 15 min. Furthermore, high activity was obtained by adjusting the concentration of the coenzyme nicotinamide adenine dinucleotide (NADH) solution added to the S-ADH-immobilized mesh to 500 μM and the solvent to a potassium phosphate buffer solution at pH 6.5. The gas imaging system using the S-ADH-immobilized mesh was able to image the decrease in NADH fluorescence (ex 340 nm, fl 490 nm) caused by the catalytic reaction of S-ADH and the acetone distribution in the concentration range of 0.1-10 ppm-v, including the breath concentration of healthy people at rest. The exhaled breath of two healthy subjects at 6 h of fasting was quantified as 377 and 673 ppb-v, which were consistent with the values quantified by gas chromatography-mass spectrometry.
{"title":"Image Sensing of Gaseous Acetone Using Secondary Alcohol Dehydrogenase-Immobilized Mesh for Exhaled Air.","authors":"Kenta Iitani, Mika Suzuki, Kenta Ichikawa, Koji Toma, Takahiro Arakawa, Kohji Mitsubayashi","doi":"10.1021/acs.analchem.4c02251","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c02251","url":null,"abstract":"<p><p>Human-borne acetone is a potent marker of lipid metabolism. Here, an enzyme immobilization method for secondary alcohol dehydrogenase (S-ADH), which is suitable for highly sensitive and selective biosensing of acetone, was developed, and then its applicability was demonstrated for spatiotemporal imaging of concentration distribution. After various investigations, S-ADH-immobilized meshes could be prepared with less than 5% variation by cross-linking S-ADH with glutaraldehyde on a cotton mesh at 40 °C for 15 min. Furthermore, high activity was obtained by adjusting the concentration of the coenzyme nicotinamide adenine dinucleotide (NADH) solution added to the S-ADH-immobilized mesh to 500 μM and the solvent to a potassium phosphate buffer solution at pH 6.5. The gas imaging system using the S-ADH-immobilized mesh was able to image the decrease in NADH fluorescence (ex 340 nm, fl 490 nm) caused by the catalytic reaction of S-ADH and the acetone distribution in the concentration range of 0.1-10 ppm-v, including the breath concentration of healthy people at rest. The exhaled breath of two healthy subjects at 6 h of fasting was quantified as 377 and 673 ppb-v, which were consistent with the values quantified by gas chromatography-mass spectrometry.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":null,"pages":null},"PeriodicalIF":6.7,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141489930","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}