Branched-chain amino acids (BCAAs) are vital for maintaining muscle performance and their measurement is critical for assessing nutritional status, health, and disease conditions. However, selective analysis of individual BCAAs remains challenging due to the limited selectivity of conventional enzymes. In this study, we explore the enzymatic detection of BCAAs (valine (Val), leucine (Leu), and isoleucine (Ile)) among 20 amino acids using their corresponding aminoacyl-tRNA synthetases (aaRSs) as recognition elements. Based on our findings, a paper-based analytical device (PAD) for Val, Leu, and Ile was developed using filtration paper.
Results
The calibration curves, and selectivity of each aaRS were assessed using a microplate reader. Trinder’s reagent was used as the colorimetric reaction following aaRS reaction. Additionally, Val, Leu, and Ile analysis was conducted on real food samples to evaluate practical applicability. Linear relationships were established for Val (2–75 μM; correlation coefficient: R2 = 0.959), Leu (2–50 μM; R2 = 0.984), and Ile (1–100 μM; R2 = 0.982), with detection limits of 18.5, 7.6, and 8.4 μM, respectively, using the microplate reader. Furthermore, PAD exhibited linear relationships for Val (7–80 μM; R2 = 0.978), Leu (9–80 μM; R2 = 0.956), and Ile (19–80 μM; R2 = 0.961), with detection limits of 2.2, 3.1, and 6.3 μM, respectively. The analysis was completed within 15 min.
Significance
Trinder’s reagent, a colorimetric reaction reagent, ensures ease of use and safety owing to its neutral pH. Furthermore, it facilitated the development of PAD and analysis of Val, Leu, and Ile. The findings of this study contribute to the advancement of robust biomedical devices precisely for amino acid analysis. High-resolution detection was accomplished using a USB camera and novel RGB value analysis equations.
{"title":"Selective measurement of valine, leucine, and isoleucine using corresponding aminoacyl-tRNA synthetases and application to paper-based analytical devices for colorimetric detection","authors":"Akimitsu Kugimiya, Makoto Kanesada, Akari Kawamura, Karen Mukainaka, Yasuhisa Nakano","doi":"10.1016/j.aca.2025.343924","DOIUrl":"https://doi.org/10.1016/j.aca.2025.343924","url":null,"abstract":"<h3>Background</h3>Branched-chain amino acids (BCAAs) are vital for maintaining muscle performance and their measurement is critical for assessing nutritional status, health, and disease conditions. However, selective analysis of individual BCAAs remains challenging due to the limited selectivity of conventional enzymes. In this study, we explore the enzymatic detection of BCAAs (valine (Val), leucine (Leu), and isoleucine (Ile)) among 20 amino acids using their corresponding aminoacyl-tRNA synthetases (aaRSs) as recognition elements. Based on our findings, a paper-based analytical device (PAD) for Val, Leu, and Ile was developed using filtration paper.<h3>Results</h3>The calibration curves, and selectivity of each aaRS were assessed using a microplate reader. Trinder’s reagent was used as the colorimetric reaction following aaRS reaction. Additionally, Val, Leu, and Ile analysis was conducted on real food samples to evaluate practical applicability. Linear relationships were established for Val (2–75 μM; correlation coefficient: <em>R</em><sup><em>2</em></sup> = 0.959), Leu (2–50 μM; <em>R</em><sup><em>2</em></sup> = 0.984), and Ile (1–100 μM; <em>R</em><sup><em>2</em></sup> = 0.982), with detection limits of 18.5, 7.6, and 8.4 μM, respectively, using the microplate reader. Furthermore, PAD exhibited linear relationships for Val (7–80 μM; <em>R</em><sup><em>2</em></sup> = 0.978), Leu (9–80 μM; <em>R</em><sup><em>2</em></sup> = 0.956), and Ile (19–80 μM; <em>R</em><sup><em>2</em></sup> = 0.961), with detection limits of 2.2, 3.1, and 6.3 μM, respectively. The analysis was completed within 15 min.<h3>Significance</h3>Trinder’s reagent, a colorimetric reaction reagent, ensures ease of use and safety owing to its neutral pH. Furthermore, it facilitated the development of PAD and analysis of Val, Leu, and Ile. The findings of this study contribute to the advancement of robust biomedical devices precisely for amino acid analysis. High-resolution detection was accomplished using a USB camera and novel RGB value analysis equations.","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"14 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143608445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-13DOI: 10.1016/j.aca.2025.343937
Thomas Swift, Edward Dyson, Natalia Koniuch, Richard Telford, Stephen Rimmer
Question
This paper studies the importance of resolving ‘in-solution’ viscosity to determine an accurate hydrodynamic radii for high molar mass or high dispersity macromolecules via DOSY NMR. Analysis of polymer size via diffusion NMR has become increasingly more common, however as in-solution viscosity increases NMR output becomes more complex and requires dedicated methodologies (both in the instrumentation and data treatment) that can sufficiently resolve slowly diffusing analytes.
Results
Diffusion measurements were used to determine hydrodynamic radii of dissolved polymer chains of materials across a broad molar mass ranges in multiple solvents. Studied systems included poly(ethylene glycol), poly(ethylene oxide), poly(styrene), poly(methyl methacrylate) and poly(N-isopropylacrylamide) and all are shown to match known literature values for dissolved polymer coils with a high degree of accuracy. However, it is shown that it is essential to use the “in-solution viscosity”, which can be obtained by applying a viscosity correction factor to the pure solvent viscosity. It was found that % error in outputs correlates to the viscosity of the solvent, with low viscosity solvents contributing to a higher variability in output data. We have also shown how the experimental range of the technique can be expanded to high molar mass (in excess of 1 million g mol-1), or high viscosity, and demonstrated the advantages of a diffusion optimised NMR probe (Bruker DiffBB) to target slowly diffusing chemical species.
Significance
The presence of even small quantities of large molar mass polymer analytes (2 mg ml-1) has an impact on in-solution viscosity, and thus provides a systematic offset in output diffusion values that are commonly used to interpret polymer sample size. DOSY NMR data include the diffusion of the solvent in-solution. Therefore, DOSY NMR measurements alone, with no internal or external standard besides the solvent itself, can be used to correct for this, allowing for prediction of an accurate hydrodynamic radius (and thus molar mass) of large, slowly diffusing, materials.
{"title":"Overcoming ‘Diffusion Limits’ – Principles Required to Measure High Molar Mass Polymers by Diffusion Ordered NMR","authors":"Thomas Swift, Edward Dyson, Natalia Koniuch, Richard Telford, Stephen Rimmer","doi":"10.1016/j.aca.2025.343937","DOIUrl":"https://doi.org/10.1016/j.aca.2025.343937","url":null,"abstract":"<h3>Question</h3>This paper studies the importance of resolving ‘in-solution’ viscosity to determine an accurate hydrodynamic radii for high molar mass or high dispersity macromolecules via DOSY NMR. Analysis of polymer size via diffusion NMR has become increasingly more common, however as in-solution viscosity increases NMR output becomes more complex and requires dedicated methodologies (both in the instrumentation and data treatment) that can sufficiently resolve slowly diffusing analytes.<h3>Results</h3>Diffusion measurements were used to determine hydrodynamic radii of dissolved polymer chains of materials across a broad molar mass ranges in multiple solvents. Studied systems included poly(ethylene glycol), poly(ethylene oxide), poly(styrene), poly(methyl methacrylate) and poly(<em>N</em>-isopropylacrylamide) and all are shown to match known literature values for dissolved polymer coils with a high degree of accuracy. However, it is shown that it is essential to use the “in-solution viscosity”, which can be obtained by applying a viscosity correction factor to the pure solvent viscosity. It was found that % error in outputs correlates to the viscosity of the solvent, with low viscosity solvents contributing to a higher variability in output data. We have also shown how the experimental range of the technique can be expanded to high molar mass (in excess of 1 million g mol<sup>-1</sup>), or high viscosity, and demonstrated the advantages of a diffusion optimised NMR probe (Bruker DiffBB) to target slowly diffusing chemical species.<h3>Significance</h3>The presence of even small quantities of large molar mass polymer analytes (2 mg ml<sup>-1</sup>) has an impact on in-solution viscosity, and thus provides a systematic offset in output diffusion values that are commonly used to interpret polymer sample size. DOSY NMR data include the diffusion of the solvent in-solution. Therefore, DOSY NMR measurements alone, with no internal or external standard besides the solvent itself, can be used to correct for this, allowing for prediction of an accurate hydrodynamic radius (and thus molar mass) of large, slowly diffusing, materials.","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"23 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143608436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-13DOI: 10.1016/j.aca.2025.343936
Miao Fang, Lingxia Yao, Jing Zhang, Xiaolin Hou
The release of radioactive substances into the environment, particularly the discharge of contaminated water from the Fukushima Daiichi Nuclear Power Plant, has raised public concern. 90Sr, a highly hazardous radionuclide, remains a significant challenge for accurate determination in environmental seawater due to its low concentration. In this work, an effective co-precipitation of 90Sr with SrCO3 and CaCO3 was established for pre-concentration of 90Sr from a large volume of seawater up to 45 L by using an appropriate concentration of (NH4)2CO3 instead of Na2CO3 followed by removal of calcium by hydroxide precipitation. A chemical yield of (88 ± 2)% was achieved for strontium from 45 L of seawater, and the decontamination factors for most radionuclides were higher than 104. The 90Y ingrown from 90Sr decay was used for the determination of 90Sr, and sulfate precipitation was employed to remove radionuclides of Sr, Ba, Ra, and Pb, and the Y was further purified by hydroxide precipitation. The chemical yield of Y was higher than 90% in this step, and the counting efficiency of 90Y by liquid scintillation counting reached 100%. The detection limit for 90Sr was estimated at 0.11 mBq/L for 45 L seawater, an order of magnitude lower than surface seawater levels without direct human nuclear contamination. The method was validated by the standard addition method and successfully applied to determine 90Sr in seawater collected from China seas. The developed method is simple and cost-effective compared to the reported methods and robust for routine analysis of seawater for 90Sr.
{"title":"Determination of low-level 90Sr in large volumes of seawater","authors":"Miao Fang, Lingxia Yao, Jing Zhang, Xiaolin Hou","doi":"10.1016/j.aca.2025.343936","DOIUrl":"https://doi.org/10.1016/j.aca.2025.343936","url":null,"abstract":"The release of radioactive substances into the environment, particularly the discharge of contaminated water from the Fukushima Daiichi Nuclear Power Plant, has raised public concern. <sup>90</sup>Sr, a highly hazardous radionuclide, remains a significant challenge for accurate determination in environmental seawater due to its low concentration. In this work, an effective co-precipitation of <sup>90</sup>Sr with SrCO<sub>3</sub> and CaCO<sub>3</sub> was established for pre-concentration of <sup>90</sup>Sr from a large volume of seawater up to 45 L by using an appropriate concentration of (NH<sub>4</sub>)<sub>2</sub>CO<sub>3</sub> instead of Na<sub>2</sub>CO<sub>3</sub> followed by removal of calcium by hydroxide precipitation. A chemical yield of (88 ± 2)% was achieved for strontium from 45 L of seawater, and the decontamination factors for most radionuclides were higher than 10<sup>4</sup>. The <sup>90</sup>Y ingrown from <sup>90</sup>Sr decay was used for the determination of <sup>90</sup>Sr, and sulfate precipitation was employed to remove radionuclides of Sr, Ba, Ra, and Pb, and the Y was further purified by hydroxide precipitation. The chemical yield of Y was higher than 90% in this step, and the counting efficiency of <sup>90</sup>Y by liquid scintillation counting reached 100%. The detection limit for <sup>90</sup>Sr was estimated at 0.11 mBq/L for 45 L seawater, an order of magnitude lower than surface seawater levels without direct human nuclear contamination. The method was validated by the standard addition method and successfully applied to determine <sup>90</sup>Sr in seawater collected from China seas. The developed method is simple and cost-effective compared to the reported methods and robust for routine analysis of seawater for <sup>90</sup>Sr.","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"16 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143608491","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-13DOI: 10.1016/j.aca.2025.343930
Runze Xu, Ning Liu, Zhenrun Li, Qiang Ma
In this work, a novel biosensor based on aggregation-induced luminescence (AIE) of copper-silver alloy nanoclusters (CuAg NCs) has been developed for the detection of gastric cancer marker miRNA-142-3p. The alloy nanoclusters were synthesized by doping Ag ions into Cu NCs as nano-emitters. On the one hand, the doped Ag ions induced Ag-Cu metallophilic interactions, which promoted the relaxation of excited electrons through the radiative pathway. On the other hand, the doped Ag ions can reduce the band gap between the HOMO-LUMO orbitals of the nanoclusters, which decreased the energy consumed for electronic excitation. Therefore, the luminescent signal and stability of CuAg NCs were significantly enhanced. Furthermore, the modification of the permanently positively charged polysaccharide quaternary ammonium salt (Curdlan-g-PGTMAC) on CuAg NCs induced the aggregation-induced electrochemiluminescence (AIECL) effect. The introduction of Curdlan-g-PGTMAC also accelerated the reduction of the co-interaction reagent (S2O82-), which significantly improved the ECL generation efficiency. The ECL response of the CuAg NC-based AIECL biosensor showed a good linear correlation with the concentration of miRNA-142-3p over a wide range from 1 fM to 100 nM with a detection limit as low as 8.8 fM. The ECL biosensor with high sensitivity and wide dynamic range was used for the clinical application of miRNA-142-3p in gastric cancer successfully.
{"title":"Aggregation-induced ECL strategy based on CuAg nanoclusters/Curdlan-g-PGTMAC for gastric cancer detection","authors":"Runze Xu, Ning Liu, Zhenrun Li, Qiang Ma","doi":"10.1016/j.aca.2025.343930","DOIUrl":"https://doi.org/10.1016/j.aca.2025.343930","url":null,"abstract":"In this work, a novel biosensor based on aggregation-induced luminescence (AIE) of copper-silver alloy nanoclusters (CuAg NCs) has been developed for the detection of gastric cancer marker miRNA-142-3p. The alloy nanoclusters were synthesized by doping Ag ions into Cu NCs as nano-emitters. On the one hand, the doped Ag ions induced Ag-Cu metallophilic interactions, which promoted the relaxation of excited electrons through the radiative pathway. On the other hand, the doped Ag ions can reduce the band gap between the HOMO-LUMO orbitals of the nanoclusters, which decreased the energy consumed for electronic excitation. Therefore, the luminescent signal and stability of CuAg NCs were significantly enhanced. Furthermore, the modification of the permanently positively charged polysaccharide quaternary ammonium salt (Curdlan-g-PGTMAC) on CuAg NCs induced the aggregation-induced electrochemiluminescence (AIECL) effect. The introduction of Curdlan-g-PGTMAC also accelerated the reduction of the co-interaction reagent (S<sub>2</sub>O<sub>8</sub><sup>2-</sup>), which significantly improved the ECL generation efficiency. The ECL response of the CuAg NC-based AIECL biosensor showed a good linear correlation with the concentration of miRNA-142-3p over a wide range from 1 fM to 100 nM with a detection limit as low as 8.8 fM. The ECL biosensor with high sensitivity and wide dynamic range was used for the clinical application of miRNA-142-3p in gastric cancer successfully.","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"10 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143608202","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-12DOI: 10.1016/j.aca.2025.343929
Jiahui Liang, Fei Chen, Zhihui Tian, Yan Zhang, Lei Zhang, Wangbao Yin, Liantuan Xiao, Suotang Jia
Background
Laser-Induced Breakdown Spectroscopy (LIBS) has demonstrated significant potential in microbial detection due to its rapid, non-contact, and multi-element analytical capabilities. However, remote detection is hindered by challenges such as signal attenuation and the high similarity of spectral features, which reduce classification accuracy. To address these issues, this study proposes a multi-temporal LIBS remote identification method optimized based on the plasma electron temperature coefficient of variation (CVT). By analyzing CVT values across different delay ranges, optimal time delays were selected and combined to amplify spectral differentiation, thereby improving classification performance.
Results
A coaxial-design LIBS telemetry system with adjustable focus was constructed, and multi-substrate telemetry testing was conducted on 10 common pathogenic bacteria at distances of 5 m and 10 m. By optimizing multiple temporal delays within the 100-1000 ns range, the classification performance of single-temporal, dual-temporal, and multi-temporal spectral combinations was evaluated. The results showed that the multi-temporal approach improved the classification performance across all substrates. At a distance of 5 m, a 100% identification rate was achieved for all substrates, with Precision, Recall, and F1-score all reaching 1.0. At 10 m, the identification rate for the aluminum substrate increased from 76% to 93%. In addition, the contribution of the four major elements, Ca, Na, C, and K, was found to account for up to 60% of the classification results.
Significance and novelty
It is demonstrated that the CVT-optimized multi-temporal LIBS technology effectively overcomes the signal attenuation bottleneck at long distances, significantly enhancing the robustness and analytical capability of remote microbial identification. This approach provides a novel method for remote detection in areas such as public safety, medical diagnostics, and military defense.
{"title":"Study on precise identification of remote bacterial species using multi-temporal LIBS optimized by plasma electron temperature coefficient of variation","authors":"Jiahui Liang, Fei Chen, Zhihui Tian, Yan Zhang, Lei Zhang, Wangbao Yin, Liantuan Xiao, Suotang Jia","doi":"10.1016/j.aca.2025.343929","DOIUrl":"https://doi.org/10.1016/j.aca.2025.343929","url":null,"abstract":"<h3>Background</h3>Laser-Induced Breakdown Spectroscopy (LIBS) has demonstrated significant potential in microbial detection due to its rapid, non-contact, and multi-element analytical capabilities. However, remote detection is hindered by challenges such as signal attenuation and the high similarity of spectral features, which reduce classification accuracy. To address these issues, this study proposes a multi-temporal LIBS remote identification method optimized based on the plasma electron temperature coefficient of variation (<em>CV</em><sub><em>T</em></sub>). By analyzing <em>CV</em><sub><em>T</em></sub> values across different delay ranges, optimal time delays were selected and combined to amplify spectral differentiation, thereby improving classification performance.<h3>Results</h3>A coaxial-design LIBS telemetry system with adjustable focus was constructed, and multi-substrate telemetry testing was conducted on 10 common pathogenic bacteria at distances of 5 m and 10 m. By optimizing multiple temporal delays within the 100-1000 ns range, the classification performance of single-temporal, dual-temporal, and multi-temporal spectral combinations was evaluated. The results showed that the multi-temporal approach improved the classification performance across all substrates. At a distance of 5 m, a 100% identification rate was achieved for all substrates, with Precision, Recall, and F1-score all reaching 1.0. At 10 m, the identification rate for the aluminum substrate increased from 76% to 93%. In addition, the contribution of the four major elements, Ca, Na, C, and K, was found to account for up to 60% of the classification results.<h3>Significance and novelty</h3>It is demonstrated that the <em>CV</em><sub><em>T</em></sub>-optimized multi-temporal LIBS technology effectively overcomes the signal attenuation bottleneck at long distances, significantly enhancing the robustness and analytical capability of remote microbial identification. This approach provides a novel method for remote detection in areas such as public safety, medical diagnostics, and military defense.","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"213 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143608493","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-11DOI: 10.1016/j.aca.2025.343931
Huanhuan Xing, Xiaojing Xing, Fangfang Chen, Ning Li, Dangdang Xu, Ruili Wu, Yanbing Lv, Lin Song Li
Background
Highly sensitive analysis of protein biomarkers with low concentrations is essential for biological research and medical diagnosis, where quantum dots (QDs) based fluorescence-linked immunoassay (QD-FLISA) has been given considerable attention among the quantitative detection due to its outstanding characteristics. However, the traditional QD-FLISA is usually subject to the low sensitivity owing to the limited photoluminescence (PL) intensity of QDs. In this sense, the development of novel strategy that could remarkably enhance the sensitive of traditional QD-FLISA would be highly desirable.
Results
Herein, DNA nanowires-mediated high sensitive QD-FLISA (DNA-nano- QD-FLISA) are first designed and used for the ultrasensitive detection of proteins, where DNA-nanowires are assembled through the hybridization chain reaction (HCR) and C-reactive protein (CRP) is chosen as the model analyte. The results demonstrate that the proposed DNA-nano-QD-FLISA can achieve sensitive detection of CRP, with a limit of detection (LOD) of 0.17 ng/mL, significantly lower than the system without DNA nanowires (1.66 ng/mL). Furthermore, the CRP levels in clinical samples were analyzed, yielding an excellent agreement with the Roche immunoturbidimetric method. Additionally, the versatility of the assay were demonstrated by adapting it to detect the other clinical proteins, interleukin-6 (IL-6) and procalcitonin (PCT), achieving the LODs of 0.07 ng/mL for IL-6 and 0.07 ng/mL for PCT. Furthermore, we found that the length of DNA nanowires significantly influenced the detection performance of QD-FLISA, offering a straightforward approach to precisely adjust the detection range.
Significance
This work presents an ultra-sensitive QD-FLISA for protein detection via the introduction of DNA-nanowires assembled through HCR. The achieved results demonstrate that the incorporation of DNA nanowires enhances the detection sensitivity and accuracy of traditional QD-FLISA in quantifying low-abundance biomarkers, which holds significant clinical importance for early disease screening and diagnosis.
{"title":"DNA nanowires-mediated high sensitive quantum dot-fluorescence-linked immunoassay for proteins analysis","authors":"Huanhuan Xing, Xiaojing Xing, Fangfang Chen, Ning Li, Dangdang Xu, Ruili Wu, Yanbing Lv, Lin Song Li","doi":"10.1016/j.aca.2025.343931","DOIUrl":"https://doi.org/10.1016/j.aca.2025.343931","url":null,"abstract":"<h3>Background</h3>Highly sensitive analysis of protein biomarkers with low concentrations is essential for biological research and medical diagnosis, where quantum dots (QDs) based fluorescence-linked immunoassay (QD-FLISA) has been given considerable attention among the quantitative detection due to its outstanding characteristics. However, the traditional QD-FLISA is usually subject to the low sensitivity owing to the limited photoluminescence (PL) intensity of QDs. In this sense, the development of novel strategy that could remarkably enhance the sensitive of traditional QD-FLISA would be highly desirable.<h3>Results</h3>Herein, DNA nanowires-mediated high sensitive QD-FLISA (DNA-nano- QD-FLISA) are first designed and used for the ultrasensitive detection of proteins, where DNA-nanowires are assembled through the hybridization chain reaction (HCR) and C-reactive protein (CRP) is chosen as the model analyte. The results demonstrate that the proposed DNA-nano-QD-FLISA can achieve sensitive detection of CRP, with a limit of detection (LOD) of 0.17 ng/mL, significantly lower than the system without DNA nanowires (1.66 ng/mL). Furthermore, the CRP levels in clinical samples were analyzed, yielding an excellent agreement with the Roche immunoturbidimetric method. Additionally, the versatility of the assay were demonstrated by adapting it to detect the other clinical proteins, interleukin-6 (IL-6) and procalcitonin (PCT), achieving the LODs of 0.07 ng/mL for IL-6 and 0.07 ng/mL for PCT. Furthermore, we found that the length of DNA nanowires significantly influenced the detection performance of QD-FLISA, offering a straightforward approach to precisely adjust the detection range.<h3>Significance</h3>This work presents an ultra-sensitive QD-FLISA for protein detection <em>via</em> the introduction of DNA-nanowires assembled through HCR. The achieved results demonstrate that the incorporation of DNA nanowires enhances the detection sensitivity and accuracy of traditional QD-FLISA in quantifying low-abundance biomarkers, which holds significant clinical importance for early disease screening and diagnosis.","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"40 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143599097","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-11DOI: 10.1016/j.aca.2025.343928
Eun Yeong Lee, Minju Lee, Myoung Gyu Kim, Chae Eun Bae, Sung-Han Kim, Yong Shin
Background
Despite significant advancements in nucleic acid testing technologies, current nucleic acid extraction methods are often limited by inefficiency, complexity, and a lack of versatility. To overcome these challenges, we have developed an innovative solid-phase extraction (SPE) method employing sulfuric acid-activated bentonite (SAB) for extracting nucleic acids from various sample types.
Results
Activation with sulfuric acid expands the surface area of bentonite by 2.2 times, thereby enhancing its adsorption capacity and surface modification efficiency. To further improve extraction efficiency, we modified SAB through amine-functionalization using 3-aminopropyl(diethoxy)methylsilane (APDMS), resulting in the creation of APDMS-modified SAB (ASAB). This modification facilitates efficient nucleic acid binding via reversible interactions mediated by a homobifunctional imidoester (HI) reagent. Our ASAB-based SPE system offers a streamlined, universal protocol for isolating DNA, RNA, and miRNA from diverse samples, including clinical bodily fluids and culture media, in under 30 minutes. Moreover, the system effectively enriches low concentrations of negatively charged pathogens (down to 20 CFU/reaction) from large-volume samples (up to 50 mL) through a 30-minute pre-enrichment step utilizing the positively charged ASAB-HI complex. Comparative testing with pooled human urine and plasma samples revealed up to a 3.95-fold increase in DNA recovery compared to commercial SPE kits. Additionally, the system demonstrated up to a 6.3-fold improvement in the isolation of unstable viral RNA from clinical nasopharyngeal swabs, as well as critical microRNA biomarkers.
Significance
The versatility and high efficiency of nucleic acid recovery with our ASAB-based SPE system indicate its potential to revolutionize traditional SPE methods, positioning it as a universal nucleic acid extraction platform for molecular biology research.
{"title":"Acid-activated bentonite for solid-phase nucleic acid extraction from various pathogenic samples","authors":"Eun Yeong Lee, Minju Lee, Myoung Gyu Kim, Chae Eun Bae, Sung-Han Kim, Yong Shin","doi":"10.1016/j.aca.2025.343928","DOIUrl":"https://doi.org/10.1016/j.aca.2025.343928","url":null,"abstract":"<h3>Background</h3>Despite significant advancements in nucleic acid testing technologies, current nucleic acid extraction methods are often limited by inefficiency, complexity, and a lack of versatility. To overcome these challenges, we have developed an innovative solid-phase extraction (SPE) method employing sulfuric acid-activated bentonite (SAB) for extracting nucleic acids from various sample types.<h3>Results</h3>Activation with sulfuric acid expands the surface area of bentonite by 2.2 times, thereby enhancing its adsorption capacity and surface modification efficiency. To further improve extraction efficiency, we modified SAB through amine-functionalization using 3-aminopropyl(diethoxy)methylsilane (APDMS), resulting in the creation of APDMS-modified SAB (ASAB). This modification facilitates efficient nucleic acid binding via reversible interactions mediated by a homobifunctional imidoester (HI) reagent. Our ASAB-based SPE system offers a streamlined, universal protocol for isolating DNA, RNA, and miRNA from diverse samples, including clinical bodily fluids and culture media, in under 30 minutes. Moreover, the system effectively enriches low concentrations of negatively charged pathogens (down to 20 CFU/reaction) from large-volume samples (up to 50 mL) through a 30-minute pre-enrichment step utilizing the positively charged ASAB-HI complex. Comparative testing with pooled human urine and plasma samples revealed up to a 3.95-fold increase in DNA recovery compared to commercial SPE kits. Additionally, the system demonstrated up to a 6.3-fold improvement in the isolation of unstable viral RNA from clinical nasopharyngeal swabs, as well as critical microRNA biomarkers.<h3>Significance</h3>The versatility and high efficiency of nucleic acid recovery with our ASAB-based SPE system indicate its potential to revolutionize traditional SPE methods, positioning it as a universal nucleic acid extraction platform for molecular biology research.","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"2 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143599096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
One of the crucial enzymes for cancer cell growth is lactate dehydrogenase (LDH, E.C. 1.1.1.27), an oxidoreductase that catalyzes the conversion between pyruvate and lactate. It has been found that in cancer cells metabolism, the LDH isoenzyme profile changes, with forms rich in the muscle-type subunit beginning to dominate over those in which the heart-type predominates. This suggests that by examining changes in the enzymatic activity of isoforms with a specific subunit content, it may be possible to quickly distinguish a physiological sample from a pathological one.
Results
This article focuses on the development of an analytical strategy that enables the estimation of the ratio of LDH fraction activities as a basis for a simple and quick screening test. Spectrophotometric detection of LDH activity is based on the ferrozine photometric reaction with ferrous ions generated during the biocatalytic reduction of ferric ions by NADH. The developed Multicommutated Flow Analysis (MCFA) system, coupled with an optoelectronic flow-through detector, enables the use of a kinetic method based on the inhibition of LDH subunits to monitor the enzyme reaction kinetics. The distinctly different responses of the muscle-type and heart-type subunits to the selected inhibitors revealed a linear relationship between the obtained analytical signal and the percentage content of each subunit. The calibration curves for selected inhibitors are linear within the tested range of standards with coefficients of determination equal to 0.99 each.
Significance
The developed MCFA system was utilized in the analysis of human serum samples obtained from both healthy patients and patients with cancer. The analysis demonstrates that the proposed approach can differentiate oncological serum samples from reference ones based on the LDH fractions activity ratio, even when their total LDH activity level is low.
{"title":"Analytical Approach to Distinguishing Lactate Dehydrogenase Fractions for Oncological Diagnostics","authors":"Justyna GŁOWACKA-GUDANEK, Kamil GRYCKIEWICZ, Kamil STRZELAK","doi":"10.1016/j.aca.2025.343921","DOIUrl":"https://doi.org/10.1016/j.aca.2025.343921","url":null,"abstract":"<h3>Background</h3>One of the crucial enzymes for cancer cell growth is lactate dehydrogenase (LDH, E.C. 1.1.1.27), an oxidoreductase that catalyzes the conversion between pyruvate and lactate. It has been found that in cancer cells metabolism, the LDH isoenzyme profile changes, with forms rich in the muscle-type subunit beginning to dominate over those in which the heart-type predominates. This suggests that by examining changes in the enzymatic activity of isoforms with a specific subunit content, it may be possible to quickly distinguish a physiological sample from a pathological one.<h3>Results</h3>This article focuses on the development of an analytical strategy that enables the estimation of the ratio of LDH fraction activities as a basis for a simple and quick screening test. Spectrophotometric detection of LDH activity is based on the ferrozine photometric reaction with ferrous ions generated during the biocatalytic reduction of ferric ions by NADH. The developed Multicommutated Flow Analysis (MCFA) system, coupled with an optoelectronic flow-through detector, enables the use of a kinetic method based on the inhibition of LDH subunits to monitor the enzyme reaction kinetics. The distinctly different responses of the muscle-type and heart-type subunits to the selected inhibitors revealed a linear relationship between the obtained analytical signal and the percentage content of each subunit. The calibration curves for selected inhibitors are linear within the tested range of standards with coefficients of determination equal to 0.99 each.<h3>Significance</h3>The developed MCFA system was utilized in the analysis of human serum samples obtained from both healthy patients and patients with cancer. The analysis demonstrates that the proposed approach can differentiate oncological serum samples from reference ones based on the LDH fractions activity ratio, even when their total LDH activity level is low.","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"54 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143590170","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Increasing attention has been caught by the allergy-related food safety issue. The rapid and sensitive diagnosing approaches are still in high demand for providing clinical reference. Paper-based analytical devices (PADs) are appealing candidates for allergy diagnosis and prediction due to their portability, stability, and operational easiness. However, the sensitivity of PADs needs to be further improved for the targets with low abundance. In addition to the complex signal amplifications, an alternative strategy that requires fewer reagents, steps, and shorter time is anticipated. (82)
Results
We report fluorescent PADs (FPADs) that can accumulate and detect the major peanut allergen glycoprotein Arachis hypogaea h2 (Ara h2)-specific IgE (sIgE). The FPADs are constructed by in-situ synthesis of blue-emissive carbon dots (BCDs) on the surface of cellulose paper, followed by the conjugation of Ara h2. After the capture of sIgE, a green-emissive carbon dots-labeled secondary anti-sIgE reporter (Ab2-GCDs) is assembled on FPADs. The detection relies on the sIgE concentration-dependent color variation of FPADs. In addition, the accumulation of sIgE is achievable by filtering the sample through FPADs, improving the assay sensitivity and efficiency. It is demonstrated that the limit of detection (LOD) is 15.7 ng/mL, evidently lower than the simple immersion-based assay (90.2 ng/mL). The excellent selectivity allows sIgE quantification in serum with high accuracy. (130)
Significance
By harnessing the outperforming sensing performance of the proposed FPADs, the rapid, accurate, and cost-efficient diagnosis and prediction of peanut allergy can be realized. In addition, the FPADs could serve as a universal sensing platform for varying targets by flexibly engineering the capture moieties on the surface of fluorescent paper. (50)
{"title":"A dual-functional paper-based analytical device for ultrasensitive detection of peanut allergen-specific IgE","authors":"Ze-Nan Ma, Jun-Jie Ding, Xin-Qiao Shi, Ying Yuan, Meng-Tian Wang, Li-Na Yu, Xiao-Jun Wang, Peng Shen","doi":"10.1016/j.aca.2025.343922","DOIUrl":"https://doi.org/10.1016/j.aca.2025.343922","url":null,"abstract":"<h3>Background</h3>Increasing attention has been caught by the allergy-related food safety issue. The rapid and sensitive diagnosing approaches are still in high demand for providing clinical reference. Paper-based analytical devices (PADs) are appealing candidates for allergy diagnosis and prediction due to their portability, stability, and operational easiness. However, the sensitivity of PADs needs to be further improved for the targets with low abundance. In addition to the complex signal amplifications, an alternative strategy that requires fewer reagents, steps, and shorter time is anticipated. (82)<h3>Results</h3>We report fluorescent PADs (FPADs) that can accumulate and detect the major peanut allergen glycoprotein <em>Arachis hypogaea</em> h2 (Ara h2)-specific IgE (sIgE). The FPADs are constructed by in-situ synthesis of blue-emissive carbon dots (BCDs) on the surface of cellulose paper, followed by the conjugation of Ara h2. After the capture of sIgE, a green-emissive carbon dots-labeled secondary anti-sIgE reporter (Ab2-GCDs) is assembled on FPADs. The detection relies on the sIgE concentration-dependent color variation of FPADs. In addition, the accumulation of sIgE is achievable by filtering the sample through FPADs, improving the assay sensitivity and efficiency. It is demonstrated that the limit of detection (LOD) is 15.7 ng/mL, evidently lower than the simple immersion-based assay (90.2 ng/mL). The excellent selectivity allows sIgE quantification in serum with high accuracy. (130)<h3>Significance</h3>By harnessing the outperforming sensing performance of the proposed FPADs, the rapid, accurate, and cost-efficient diagnosis and prediction of peanut allergy can be realized. In addition, the FPADs could serve as a universal sensing platform for varying targets by flexibly engineering the capture moieties on the surface of fluorescent paper. (50)","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"73 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143599634","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Fluorescence imaging agents have been benefiting tremendously from tailor-made aggregation-induced emission (AIE) luminogens, owing to their high on-off ratio, large signal contrast, low background noise as well as the resistance to photobleaching. In the domain of fingerprint imaging, AIE luminogens are beginning to exhibit an advantage owing to the aforementioned superiorities.
Results
We present an amphiphilic benzoic-acid salicylaldehyde AIE luminogen AIE-BASB, and outline its water sensitivity, self-assembly behavior as well as fingerprint imaging properties. AIE-BASB self-assembles into nanoscale textures when fabricated into a drop-casting film but undergoes a disassembly process in response to trace water on fingertip upon physical-contacting. Owing to the biological textures on the skin, fingerprint image can be clearly recorded by AIE-BASB film, which reveals detailed microscopic features of fingerprint information ranging from level-1 to level-3. Furthermore, it allows accurate measurements of the sizes, shapes, centroids, and areas of the sweat pores, which leads the fingerprint information into the next level. In addition, we develop an intelligent system based on AIE-BASB by integrating hardware and software modules, which is capable of recording and identifying fingerprint. After inputting fingerprint segments in trial operation, this intelligent system makes identification by calculation of the categorical probability, and successfully predicts the classification of the undefined fingerprint segments, implying 100% accuracy in fingerprint identification.
Significance
We predict that AIE-BASB may facilitate the development of new biometric technologies, which have broad applications in the domain of artificial intelligence, including machine tactility, target perception and object-machine interaction.
{"title":"Mapping human fingerprint beyond level-3 based on an amphiphilic aggregation-induced emission luminogen and the construction of intelligent platform for personal identification","authors":"Xinyi Zhao, Zixuan Wang, Haoran Liu, Siyu Yan, Zihan Liu, Yuai Duan, Tianyu Han, Tiandong Han","doi":"10.1016/j.aca.2025.343927","DOIUrl":"https://doi.org/10.1016/j.aca.2025.343927","url":null,"abstract":"<h3>Background</h3>Fluorescence imaging agents have been benefiting tremendously from tailor-made aggregation-induced emission (AIE) luminogens, owing to their high on-off ratio, large signal contrast, low background noise as well as the resistance to photobleaching. In the domain of fingerprint imaging, AIE luminogens are beginning to exhibit an advantage owing to the aforementioned superiorities.<h3>Results</h3>We present an amphiphilic benzoic-acid salicylaldehyde AIE luminogen <strong>AIE-BASB</strong>, and outline its water sensitivity, self-assembly behavior as well as fingerprint imaging properties. <strong>AIE-BASB</strong> self-assembles into nanoscale textures when fabricated into a drop-casting film but undergoes a disassembly process in response to trace water on fingertip upon physical-contacting. Owing to the biological textures on the skin, fingerprint image can be clearly recorded by <strong>AIE-BASB</strong> film, which reveals detailed microscopic features of fingerprint information ranging from level-1 to level-3. Furthermore, it allows accurate measurements of the sizes, shapes, centroids, and areas of the sweat pores, which leads the fingerprint information into the next level. In addition, we develop an intelligent system based on <strong>AIE-BASB</strong> by integrating hardware and software modules, which is capable of recording and identifying fingerprint. After inputting fingerprint segments in trial operation, this intelligent system makes identification by calculation of the categorical probability, and successfully predicts the classification of the undefined fingerprint segments, implying 100% accuracy in fingerprint identification.<h3>Significance</h3>We predict that <strong>AIE-BASB</strong> may facilitate the development of new biometric technologies, which have broad applications in the domain of artificial intelligence, including machine tactility, target perception and object-machine interaction.","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"68 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143590241","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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