Pub Date : 2025-01-13DOI: 10.1021/acs.analchem.4c04414
Bin Wu, Xingliang He, Xing Guo, Fulong Deng, Hongen Sun, Yi Pan, Yixiang Duan, Zhongjun Zhao
The development of an atmospheric pressure interface (API) with a high ion transfer efficiency and wide mass range is advantageous for the performance improvement of mass spectrometry (MS) instruments. In this work, a novel ion guide, namely, the double-helix electrode ion funnel (DHE-IF), has been developed to enhance the ion transmission over a wide mass range in the rough vacuum region. The DHE-IF consists of two funnel-shaped helix electrodes. There are almost no potential “traps” along the central axis of DHE-IF due to the continuous double-helix electrode structure compared to the stacked ring ion funnel. The electrode design of the DHE-IF assembly was guided by ion trajectory simulations. After being fabricated, DHE-IF was integrated into an ESI-TOF-MS platform for tests. A conventional stacked ring ion funnel (IF) was also tested for comparison. The experimental results showed that DHE-IF extended the transmission window of the IF and improved the efficiency of the simultaneous transferring of low and medium m/z ions. In addition, the intensities of caffeine ions (m/z = 195) and reserpine ions (m/z = 609) were enhanced by more than 50% and 10%, respectively. These values were compared with the results obtained by the IF. The DHE-IF is expected to be widely used as an ion import device in MS instruments, which is due to its improved performance and advantages in, e.g., integration and power supply design.
{"title":"Double-Helix Electrode Ion Funnel: A New Ion Funnel Design with an Extended Mass Range","authors":"Bin Wu, Xingliang He, Xing Guo, Fulong Deng, Hongen Sun, Yi Pan, Yixiang Duan, Zhongjun Zhao","doi":"10.1021/acs.analchem.4c04414","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c04414","url":null,"abstract":"The development of an atmospheric pressure interface (API) with a high ion transfer efficiency and wide mass range is advantageous for the performance improvement of mass spectrometry (MS) instruments. In this work, a novel ion guide, namely, the double-helix electrode ion funnel (DHE-IF), has been developed to enhance the ion transmission over a wide mass range in the rough vacuum region. The DHE-IF consists of two funnel-shaped helix electrodes. There are almost no potential “traps” along the central axis of DHE-IF due to the continuous double-helix electrode structure compared to the stacked ring ion funnel. The electrode design of the DHE-IF assembly was guided by ion trajectory simulations. After being fabricated, DHE-IF was integrated into an ESI-TOF-MS platform for tests. A conventional stacked ring ion funnel (IF) was also tested for comparison. The experimental results showed that DHE-IF extended the transmission window of the IF and improved the efficiency of the simultaneous transferring of low and medium <i>m</i>/<i>z</i> ions. In addition, the intensities of caffeine ions (<i>m</i>/<i>z</i> = 195) and reserpine ions (<i>m</i>/<i>z</i> = 609) were enhanced by more than 50% and 10%, respectively. These values were compared with the results obtained by the IF. The DHE-IF is expected to be widely used as an ion import device in MS instruments, which is due to its improved performance and advantages in, e.g., integration and power supply design.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"6 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142968376","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 : 2025-01-13DOI: 10.1021/acs.analchem.4c05296
Haiyan Li, Xin Wang, Yafei Chen, Han Leng, Junjie Bai, Yi Lu, Nan Wang, Cheng Du, Jianhua Wang, Mingli Chen
The advancement of lanthanide fingerprint sensors characterized by targeted emission responses and low self-fluorescence interference for the detection of biothiols is of considerable importance for the early diagnosis and treatment of cancer. Herein, the lanthanide “personality function tailoring” HOF composite sensor array is designed for the specific discrimination of biothiols (GSH, Cys, and Hcy) based on the activation of various luminescent molecules, such as r-AuNCs/luminol via HOF surface proximity. Lumi-HOF@Ce serves as a versatile platform for catalyzing the oxidation of o-phenylenediamine (OPD) to generate yellow fluorescent oligomers, accompanied by the fluorescence attenuation of luminol. HOF@Tb functions as a confinement interface that gathers gold nanoclusters (r-AuNCs) with red fluorescence, facilitating an aggregation-induced emission enhancement (AIEE). The fluorescence properties of AuNCs are subsequently impacted to varying degrees by the Au(I)-thiolate motifs from biothiol rooted in an enhanced ligand–metal charge transfer (LMCT) process. Additionally, the catalytic activity of Lumi-HOF@Ce, which exhibits oxidase-like properties, can be inhibited by different biothiols to varying extents. The five-channel fluorescent array demonstrates exceptional discrimination of biothiol fingerprints, aided by machine learning algorithms. Feature-tailored lanthanide HOF sensor arrays achieve sensitive identification with nearly 100% accuracy in classifying clinical liver cancer samples versus normal samples, using a logic gate strategy. The current strategy of lanthanide function tailoring boosts the suitability of biosensing applications.
{"title":"Lanthanide-Assisted Function Tailoring of the HOF-Based Logic Gate Sensor Array for Biothiol Detection and Disease Discrimination","authors":"Haiyan Li, Xin Wang, Yafei Chen, Han Leng, Junjie Bai, Yi Lu, Nan Wang, Cheng Du, Jianhua Wang, Mingli Chen","doi":"10.1021/acs.analchem.4c05296","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c05296","url":null,"abstract":"The advancement of lanthanide fingerprint sensors characterized by targeted emission responses and low self-fluorescence interference for the detection of biothiols is of considerable importance for the early diagnosis and treatment of cancer. Herein, the lanthanide “personality function tailoring” HOF composite sensor array is designed for the specific discrimination of biothiols (GSH, Cys, and Hcy) based on the activation of various luminescent molecules, such as r-AuNCs/luminol via HOF surface proximity. Lumi-HOF@Ce serves as a versatile platform for catalyzing the oxidation of <i>o</i>-phenylenediamine (OPD) to generate yellow fluorescent oligomers, accompanied by the fluorescence attenuation of luminol. HOF@Tb functions as a confinement interface that gathers gold nanoclusters (r-AuNCs) with red fluorescence, facilitating an aggregation-induced emission enhancement (AIEE). The fluorescence properties of AuNCs are subsequently impacted to varying degrees by the Au(I)-thiolate motifs from biothiol rooted in an enhanced ligand–metal charge transfer (LMCT) process. Additionally, the catalytic activity of Lumi-HOF@Ce, which exhibits oxidase-like properties, can be inhibited by different biothiols to varying extents. The five-channel fluorescent array demonstrates exceptional discrimination of biothiol fingerprints, aided by machine learning algorithms. Feature-tailored lanthanide HOF sensor arrays achieve sensitive identification with nearly 100% accuracy in classifying clinical liver cancer samples versus normal samples, using a logic gate strategy. The current strategy of lanthanide function tailoring boosts the suitability of biosensing applications.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"58 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142968423","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}
Single-cell metabolic analysis has not yet achieved the coverage of bulk analysis due to the diversity of cellular metabolites and the ionization competition among species. Direct ionization methods without separation lead to the masking of low-intensity species. By designing a capillary column emitter and introducing reverse-phase chromatography principles, we achieved the microseparation of lipophilic and hydrophilic metabolites and lowered the limit of detection of hydrophilic metabolites to the level of a single oocyte. We identified 517 metabolite species in a single oocyte, achieving coverage and reproducibility comparable to those of bulk analysis. By comparing oocytes at different maturation stages, 76 metabolic features were identified with significant differences between the germinal vesicle and meiosis II stages. Metabolite level changes suggested the roles of lipid metabolism remodeling, increased amino acid synthesis, and a shift from pyrimidine metabolism to purine metabolism in the process of oocyte maturation. This microseparation mass spectrometry analysis is expected to promote single-cell metabolomics.
{"title":"Microseparation of Lipophilic and Hydrophilic Metabolites for Single Oocyte Mass Spectrometry Analysis","authors":"Jinlei Yang, Jing Wang, Runsong Cheng, Tianyi Liao, Siyuan Pan, Murong Du, Weiliang Liu, Liying Yan, Sichun Zhang, Xinrong Zhang","doi":"10.1021/acs.analchem.4c04900","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c04900","url":null,"abstract":"Single-cell metabolic analysis has not yet achieved the coverage of bulk analysis due to the diversity of cellular metabolites and the ionization competition among species. Direct ionization methods without separation lead to the masking of low-intensity species. By designing a capillary column emitter and introducing reverse-phase chromatography principles, we achieved the microseparation of lipophilic and hydrophilic metabolites and lowered the limit of detection of hydrophilic metabolites to the level of a single oocyte. We identified 517 metabolite species in a single oocyte, achieving coverage and reproducibility comparable to those of bulk analysis. By comparing oocytes at different maturation stages, 76 metabolic features were identified with significant differences between the germinal vesicle and meiosis II stages. Metabolite level changes suggested the roles of lipid metabolism remodeling, increased amino acid synthesis, and a shift from pyrimidine metabolism to purine metabolism in the process of oocyte maturation. This microseparation mass spectrometry analysis is expected to promote single-cell metabolomics.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"16 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142968414","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 : 2025-01-12DOI: 10.1021/acs.analchem.4c05401
Jakub Máčala, Saara Kuusinen, Satu Lahtinen, Hans H. Gorris, Petr Skládal, Zdeněk Farka, Tero Soukka
The anti-Stokes emission of photon upconversion nanoparticles (UCNPs) facilitates their use as labels for ultrasensitive detection in biological samples as infrared excitation does not induce autofluorescence at visible wavelengths. The detection of extremely low-abundance analytes, however, remains challenging as it is impossible to completely avoid nonspecific binding of label conjugates. To overcome this limitation, we developed a novel hybridization complex transfer technique using UCNP labels to detect short nucleic acids directly without target amplification. The assay involves capturing the target–label complexes on an initial solid phase, then using releasing oligonucleotides to specifically elute only the target–UCNP complexes and recapturing them on another solid phase. The nonspecifically adsorbed labels remain on the first solid phase, enabling background-free, ultrasensitive detection. When magnetic microparticles were used as the first solid phase in a sample volume of 120 μL, the assay achieved a limit of detection (LOD) of 310 aM, a 27-fold improvement over the reference assay without transfer. Moreover, the additional target-specific steps introduced in the complex transfer procedure improved the sequence specificity of the complex transfer assay compared with the reference assay. The suitability for clinical analysis was confirmed using spiked plasma samples, resulting in an LOD of 190 aM. By increasing the sample volume to 600 μL and using magnetic preconcentration, the LOD was improved to 46 aM. These results highlight the importance of background elimination in achieving ultralow LODs for the analysis of low-abundance biomarkers.
{"title":"Amplification-Free Attomolar Detection of Short Nucleic Acids with Upconversion Luminescence: Eliminating Nonspecific Binding by Hybridization Complex Transfer","authors":"Jakub Máčala, Saara Kuusinen, Satu Lahtinen, Hans H. Gorris, Petr Skládal, Zdeněk Farka, Tero Soukka","doi":"10.1021/acs.analchem.4c05401","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c05401","url":null,"abstract":"The anti-Stokes emission of photon upconversion nanoparticles (UCNPs) facilitates their use as labels for ultrasensitive detection in biological samples as infrared excitation does not induce autofluorescence at visible wavelengths. The detection of extremely low-abundance analytes, however, remains challenging as it is impossible to completely avoid nonspecific binding of label conjugates. To overcome this limitation, we developed a novel hybridization complex transfer technique using UCNP labels to detect short nucleic acids directly without target amplification. The assay involves capturing the target–label complexes on an initial solid phase, then using releasing oligonucleotides to specifically elute only the target–UCNP complexes and recapturing them on another solid phase. The nonspecifically adsorbed labels remain on the first solid phase, enabling background-free, ultrasensitive detection. When magnetic microparticles were used as the first solid phase in a sample volume of 120 μL, the assay achieved a limit of detection (LOD) of 310 aM, a 27-fold improvement over the reference assay without transfer. Moreover, the additional target-specific steps introduced in the complex transfer procedure improved the sequence specificity of the complex transfer assay compared with the reference assay. The suitability for clinical analysis was confirmed using spiked plasma samples, resulting in an LOD of 190 aM. By increasing the sample volume to 600 μL and using magnetic preconcentration, the LOD was improved to 46 aM. These results highlight the importance of background elimination in achieving ultralow LODs for the analysis of low-abundance biomarkers.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"1 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142962755","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}
A novel sensing platform was constructed for the recognition and identification of dihydroxybenzene isomers based on the MOF-0.02TEA fluorescence sensor with the morphology of nanosheet microspheres through coordination modulation. Based on the sensing principle that the amino group on the MOF-0.02TEA can make the Michael reaction with o-benzoquinone and p-benzoquinone, which were individually the oxidation intermediate of catechol and hydroquinone, the fluorescence intensity of MOF-0.02TEA could be quenched through the inner filter effect (IFE) without the interference from resorcinol. Besides, catechol and hydroquinone could be further distinguished with the assistance of the Schiff-base reaction by introducing o-phenylenediamine (OPD) to the detection system. The MOF-0.02TEA sensor exhibited good selectivity, and the detection limits for catechol and hydroquinone were 90.5 nmol/L and 0.52 μmol/L (S/N = 3), respectively. Moreover, the sensor could be used for the determination of dihydroxybenzene isomers in tap water and lake water.
{"title":"Michael and Schiff-Base Reactions-Assisted Fluorescence Sensor Based on the MOF Nanosheet Microspheres for the Effective Discrimination and Detection of Hydroquinone and Catechol","authors":"Jiayi Fan, Jinfan Li, Chunyan Liu, Mengyun Lu, Xinwen Jia, Wuduo Zhao, Ajuan Yu, Shusheng Zhang","doi":"10.1021/acs.analchem.4c06359","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c06359","url":null,"abstract":"A novel sensing platform was constructed for the recognition and identification of dihydroxybenzene isomers based on the MOF-0.02TEA fluorescence sensor with the morphology of nanosheet microspheres through coordination modulation. Based on the sensing principle that the amino group on the MOF-0.02TEA can make the Michael reaction with <i>o</i>-benzoquinone and <i>p</i>-benzoquinone, which were individually the oxidation intermediate of catechol and hydroquinone, the fluorescence intensity of MOF-0.02TEA could be quenched through the inner filter effect (IFE) without the interference from resorcinol. Besides, catechol and hydroquinone could be further distinguished with the assistance of the Schiff-base reaction by introducing <i>o</i>-phenylenediamine (OPD) to the detection system. The MOF-0.02TEA sensor exhibited good selectivity, and the detection limits for catechol and hydroquinone were 90.5 nmol/L and 0.52 μmol/L (S/N = 3), respectively. Moreover, the sensor could be used for the determination of dihydroxybenzene isomers in tap water and lake water.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"40 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142961345","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 : 2025-01-11DOI: 10.1021/acs.analchem.4c03660
Matthew J. Herman, Chris E. Freye
Alteration analysis (ALA), an unsupervised chemometric technique, was evaluated for its ability to discover statistically significant trends in chromatographic data sets. Recently introduced, adoption of ALA has been limited due to uncertainty regarding its sensitivity to minor changes, and there are no rules implementing ALA especially for multivariate data sets such as liquid or gas chromatography coupled to mass spectrometry. Using in-silico data sets, ALA limits of discovery for various signal-to-noises (S/Ns), rates of change across samples, and a number of samples were assessed. For 10 samples, ALA discovered changes of ∼2% across each sample for low S/Ns (15–50), ∼1% change across each sample for moderate S/Ns (65–200), and as little as a 0.1% change at high S/Ns. ALA was also evaluated for unresolved chromatographic peaks, detecting changes down to a resolution of 0.01. In tandem with ALA, two-dimensional correlation analysis (2DCOR), a nonquantitative technique, was employed post-ALA processing to provide unique insights into the relationships between the chemical changes across simulated data sets. Finally, ALA and 2DCOR were applied to the pyrolysis gas chromatography–mass spectrometry (pyGC-MS) of Kraton G1650, a styrene-ethylene-butylene-stryrene (SEBS) polymer, pyrolyzed at temperatures ranging from 350 to 700 °C. A total of 523 statistically significant chemical compounds were discovered. The ALA output was fed into 2DCOR, and a subset of the data was evaluated to understand the relationship between the chemical changes of four selected statistically significant compounds.
{"title":"Discovery-Based Analysis for Chemical Trends in Chromatographic Data Sets Using Alteration Analysis and Two-Dimensional Correlation Analysis","authors":"Matthew J. Herman, Chris E. Freye","doi":"10.1021/acs.analchem.4c03660","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c03660","url":null,"abstract":"Alteration analysis (ALA), an unsupervised chemometric technique, was evaluated for its ability to discover statistically significant trends in chromatographic data sets. Recently introduced, adoption of ALA has been limited due to uncertainty regarding its sensitivity to minor changes, and there are no rules implementing ALA especially for multivariate data sets such as liquid or gas chromatography coupled to mass spectrometry. Using in-silico data sets, ALA limits of discovery for various signal-to-noises (S/Ns), rates of change across samples, and a number of samples were assessed. For 10 samples, ALA discovered changes of ∼2% across each sample for low S/Ns (15–50), ∼1% change across each sample for moderate S/Ns (65–200), and as little as a 0.1% change at high S/Ns. ALA was also evaluated for unresolved chromatographic peaks, detecting changes down to a resolution of 0.01. In tandem with ALA, two-dimensional correlation analysis (2DCOR), a nonquantitative technique, was employed post-ALA processing to provide unique insights into the relationships between the chemical changes across simulated data sets. Finally, ALA and 2DCOR were applied to the pyrolysis gas chromatography–mass spectrometry (pyGC-MS) of Kraton G1650, a styrene-ethylene-butylene-stryrene (SEBS) polymer, pyrolyzed at temperatures ranging from 350 to 700 °C. A total of 523 statistically significant chemical compounds were discovered. The ALA output was fed into 2DCOR, and a subset of the data was evaluated to understand the relationship between the chemical changes of four selected statistically significant compounds.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"9 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142961347","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 : 2025-01-11DOI: 10.1021/acs.analchem.4c05830
Lu huang, Hong Jiang, Bin Li, Lanying Chen, Tongtong Yang, Jiang Wang, Huanwen Chen
An online reactive internal extraction electrospray ionization (iEESI) method was developed for the rapid determination of organic and inorganic speciation information for selenium in poultry tissue samples without complex sample pretreatment. The addition of citric acid as a reducing agent to the internal extraction solvent of methanol/acetic acid (99:1, V/V) for iEESI resulted in the reduction of selenate in the sample to selenite, accompanied by the production of malic acid as an oxidation product. The quantitative analysis of selenate was conducted by using malic acid. By addition of o-phenylenediamine (OPD) to the extractant, inorganic selenium including selenite and selenite reduced from selenate was immediately extracted and converted into 1,3-dihydro-2,1,3-benzoselenadiazole after an in situ reaction. The methanol in the reagent extracted organic selenium compounds from the samples, enabling a rapid quantitative analysis of organic and inorganic selenium in chicken tissue. The concentrations of selenomethionine (SeMet), l-selenocystine (SeCys(2)), selenite, and selenate in different tissue samples were examined, showing strong linearity in the range of 1.00–100.00 μg/kg with the limit of detection (LOD) ranging from 0.36 to 0.84 μg/kg. The results were validated by two conventional methods with an accuracy of 84.2–107.6%. The quantitative analysis of three chicken visceral samples revealed that the highest concentration of organic selenium was present in chicken livers, while the highest concentration of inorganic selenium was observed in gizzards. The findings may provide a reliable method for studying selenium metabolism pathways in organisms.
{"title":"Rapid Determination of Organic and Inorganic Selenium in Poultry Tissues by Internal Extractive Electrospray Ionization Mass Spectrometry","authors":"Lu huang, Hong Jiang, Bin Li, Lanying Chen, Tongtong Yang, Jiang Wang, Huanwen Chen","doi":"10.1021/acs.analchem.4c05830","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c05830","url":null,"abstract":"An online reactive internal extraction electrospray ionization (iEESI) method was developed for the rapid determination of organic and inorganic speciation information for selenium in poultry tissue samples without complex sample pretreatment. The addition of citric acid as a reducing agent to the internal extraction solvent of methanol/acetic acid (99:1, V/V) for iEESI resulted in the reduction of selenate in the sample to selenite, accompanied by the production of malic acid as an oxidation product. The quantitative analysis of selenate was conducted by using malic acid. By addition of <i>o</i>-phenylenediamine (OPD) to the extractant, inorganic selenium including selenite and selenite reduced from selenate was immediately extracted and converted into 1,3-dihydro-2,1,3-benzoselenadiazole after an in situ reaction. The methanol in the reagent extracted organic selenium compounds from the samples, enabling a rapid quantitative analysis of organic and inorganic selenium in chicken tissue. The concentrations of selenomethionine (SeMet), <span>l</span>-selenocystine (SeCys(2)), selenite, and selenate in different tissue samples were examined, showing strong linearity in the range of 1.00–100.00 μg/kg with the limit of detection (LOD) ranging from 0.36 to 0.84 μg/kg. The results were validated by two conventional methods with an accuracy of 84.2–107.6%. The quantitative analysis of three chicken visceral samples revealed that the highest concentration of organic selenium was present in chicken livers, while the highest concentration of inorganic selenium was observed in gizzards. The findings may provide a reliable method for studying selenium metabolism pathways in organisms.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"18 3 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142961349","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 : 2025-01-10DOI: 10.1021/acs.analchem.4c04113
Yanawut Manmana, Shuma Kinugasa, Yuki Hiruta, Daniel Citterio
The integration of barcode technology with smartphones on paper-based analytical devices (PADs) presents a promising approach to bridging manual detection with digital interpretation and data storage. However, previous studies of 1D barcode approaches have been limited to providing only a “yes/no” response for analyte detection. Herein, a method of using barcode readout for semiquantitative signal detection on PADs has been achieved through the integration of barcode technology with a distance-based measurement concept on PADs. To demonstrate the feasibility of this concept, a PAD fabrication strategy incorporating barcodes was explored, using the enzymatic reaction between horseradish peroxidase (HRP), 3,3′-diaminobenzidine (DAB), and H2O2 as a model system. The enzyme-catalyzed polymerization of DAB to polyDAB in the presence of hydrogen peroxide results in the appearance of color observable by the naked eye inside a paperfluidic channel, with the color-changed length depending on the H2O2 concentration. At the same time, the barcode pattern displayed as a result of this distance-based color evolution overlaid with a paper-based barcode layer can be read using a smartphone application. Parameters affecting the signal readout performance were studied. The developed device can be used to detect H2O2 concentrations in the range of 0.25 to 10 mM within 90 min with 79.6% of barcode signals correctly readable. Additionally, results from different smartphone models showed a consistent reading performance (78.4–79.6%). Finally, the quantification of glucose levels in artificial urine samples was demonstrated. This developed PAD signaling strategy offers end-users more simplicity and can be used as a standalone device or in conjunction with other digital devices.
{"title":"Development of a Semiquantitative Barcode Readout Approach for Paper-Based Analytical Devices (PADs) for Enzymatic H2O2 and Glucose Detection","authors":"Yanawut Manmana, Shuma Kinugasa, Yuki Hiruta, Daniel Citterio","doi":"10.1021/acs.analchem.4c04113","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c04113","url":null,"abstract":"The integration of barcode technology with smartphones on paper-based analytical devices (PADs) presents a promising approach to bridging manual detection with digital interpretation and data storage. However, previous studies of 1D barcode approaches have been limited to providing only a “yes/no” response for analyte detection. Herein, a method of using barcode readout for semiquantitative signal detection on PADs has been achieved through the integration of barcode technology with a distance-based measurement concept on PADs. To demonstrate the feasibility of this concept, a PAD fabrication strategy incorporating barcodes was explored, using the enzymatic reaction between horseradish peroxidase (HRP), 3,3′-diaminobenzidine (DAB), and H<sub>2</sub>O<sub>2</sub> as a model system. The enzyme-catalyzed polymerization of DAB to polyDAB in the presence of hydrogen peroxide results in the appearance of color observable by the naked eye inside a paperfluidic channel, with the color-changed length depending on the H<sub>2</sub>O<sub>2</sub> concentration. At the same time, the barcode pattern displayed as a result of this distance-based color evolution overlaid with a paper-based barcode layer can be read using a smartphone application. Parameters affecting the signal readout performance were studied. The developed device can be used to detect H<sub>2</sub>O<sub>2</sub> concentrations in the range of 0.25 to 10 mM within 90 min with 79.6% of barcode signals correctly readable. Additionally, results from different smartphone models showed a consistent reading performance (78.4–79.6%). Finally, the quantification of glucose levels in artificial urine samples was demonstrated. This developed PAD signaling strategy offers end-users more simplicity and can be used as a standalone device or in conjunction with other digital devices.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"1 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142940540","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 : 2025-01-10DOI: 10.1021/acs.analchem.4c04217
Jiahong Jin, Dujie Liao, Lin Zhao, Marion S. Greene, Yu Sa, Heng Hong, Xin-Hua Hu
Diffraction imaging of cells allows rapid phenotyping by the response of intracellular molecules to coherent illumination. However, its ability to distinguish numerous types of human leukocytes remains to be investigated. Here, we show that accurate classification of three lymphocyte subtypes can be achieved with features extracted from cross-polarized diffraction image (p-DI) pairs. A deep neural network (DNN) of DINet-PS has been developed for feature extraction from and filtering of, in the angular frequency domain, p-DI pairs acquired from live lymphocytes isolated from human spleen tissues. We built the network in a dual-channel structure and incorporated two adaptive spectral filter blocks to actively suppress extracted features related to the noise component of light in p-DI pairs. The DINet-PS was trained with p-DI pairs acquired from 5311 CD4+ T, 3819 CD8+ T, and 4054 CD19+ B cells after preprocessing and rebelling of manually derived secondary labels and classification accuracy of 96.6 ± 0.40% has been achieved in hold-out test data sets among the three subtypes. Our results show the power of DNN to extract cell-related features from p-DI pairs and the potential of polarization diffraction imaging flow cytometry for accurate and label-free classification of lymphocyte subtypes in particular and leukocytes in general.
{"title":"Accurate Classification of Human CD4+ T, CD8+ T, and CD19+ B Cells Isolated from Splenocytes by Cross-Polarized Diffraction Image Pairs","authors":"Jiahong Jin, Dujie Liao, Lin Zhao, Marion S. Greene, Yu Sa, Heng Hong, Xin-Hua Hu","doi":"10.1021/acs.analchem.4c04217","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c04217","url":null,"abstract":"Diffraction imaging of cells allows rapid phenotyping by the response of intracellular molecules to coherent illumination. However, its ability to distinguish numerous types of human leukocytes remains to be investigated. Here, we show that accurate classification of three lymphocyte subtypes can be achieved with features extracted from cross-polarized diffraction image (p-DI) pairs. A deep neural network (DNN) of DINet-PS has been developed for feature extraction from and filtering of, in the angular frequency domain, p-DI pairs acquired from live lymphocytes isolated from human spleen tissues. We built the network in a dual-channel structure and incorporated two adaptive spectral filter blocks to actively suppress extracted features related to the noise component of light in p-DI pairs. The DINet-PS was trained with p-DI pairs acquired from 5311 CD4+ T, 3819 CD8+ T, and 4054 CD19+ B cells after preprocessing and rebelling of manually derived secondary labels and classification accuracy of 96.6 ± 0.40% has been achieved in hold-out test data sets among the three subtypes. Our results show the power of DNN to extract cell-related features from p-DI pairs and the potential of polarization diffraction imaging flow cytometry for accurate and label-free classification of lymphocyte subtypes in particular and leukocytes in general.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"2 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142940542","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 : 2025-01-10DOI: 10.1021/acs.analchem.4c04689
Noor Ali, Swapnil Singh, Chaitrali Sengupta, Shashwati Paul, Megan C. Thielges
Two-dimensional infrared (2D IR) spectroscopy is a powerful technique for measuring molecular heterogeneity and dynamics with a high spatiotemporal resolution. The methods can be applied to characterize specific residues of proteins by incorporating frequency-resolved vibrational labels. However, the time scale of dynamics that 2D IR spectroscopy can measure is limited by the vibrational label’s excited-state lifetime due to the decay of 2D IR absorption bands. To extend this time scale, vibrational labels with longer lifetimes are sought. An effective approach to inhibiting intramolecular energy relaxation is to isolate the vibration from the rest of the molecule by inserting a heavy atom bridge. Although this strategy has been demonstrated through the generation of functionalized amino acids, a straightforward route to their selective incorporation into proteins is often unclear. A facile approach for the attachment of a cyano group at cysteine to generate a thiocyanate has contributed to its adoption as a vibrational label of proteins. We demonstrate that an analogous route can be used for introducing cyanoselenocysteine to generate a selenocyanate vibrational label containing a heavier bridge atom. We confirm by infrared pump–probe and 2D IR spectroscopy longer vibrational lifetimes of 100–250 ps, depending on the solvent, which enable the collection of 2D IR spectra to measure frequency dynamics on longer time scales.
{"title":"Facile Generation of Cyanoselenocysteine as a Vibrational Label for Measuring Protein Dynamics on Longer Time Scales by 2D IR Spectroscopy","authors":"Noor Ali, Swapnil Singh, Chaitrali Sengupta, Shashwati Paul, Megan C. Thielges","doi":"10.1021/acs.analchem.4c04689","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c04689","url":null,"abstract":"Two-dimensional infrared (2D IR) spectroscopy is a powerful technique for measuring molecular heterogeneity and dynamics with a high spatiotemporal resolution. The methods can be applied to characterize specific residues of proteins by incorporating frequency-resolved vibrational labels. However, the time scale of dynamics that 2D IR spectroscopy can measure is limited by the vibrational label’s excited-state lifetime due to the decay of 2D IR absorption bands. To extend this time scale, vibrational labels with longer lifetimes are sought. An effective approach to inhibiting intramolecular energy relaxation is to isolate the vibration from the rest of the molecule by inserting a heavy atom bridge. Although this strategy has been demonstrated through the generation of functionalized amino acids, a straightforward route to their selective incorporation into proteins is often unclear. A facile approach for the attachment of a cyano group at cysteine to generate a thiocyanate has contributed to its adoption as a vibrational label of proteins. We demonstrate that an analogous route can be used for introducing cyanoselenocysteine to generate a selenocyanate vibrational label containing a heavier bridge atom. We confirm by infrared pump–probe and 2D IR spectroscopy longer vibrational lifetimes of 100–250 ps, depending on the solvent, which enable the collection of 2D IR spectra to measure frequency dynamics on longer time scales.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"22 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142940543","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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