El Hadji Tombé Bodian, Coumba Faye, Diène Diégane Thiaré, Ndeye Arame Diop, Pape Abdoulaye Diaw, François Delattre, Atanasse Coly, Philippe Giamarchi
The effect of adding organized supramolecular systems such as β-cyclodextrin (β-CD) and hydroxypropyl-β-cyclodextrin (HP-β-CD) on the photochemically-induced fluorescence (PIF) spectral properties of tau-fluvalinate (TFV) in aqueous solutions was examined. The influence of pH, UV irradiation time and photoproduct stability on the cyclodextrin-enhanced photochemically-induced fluorescence intensity was also investigated. The spectral changes associated with the inclusion process yielded values for the formation constants of TFV inclusion complexes between 450 and 640 M-1, which were calculated using the nonlinear iterative regression approach least squares. In addition, host-guest interaction was clearly determined by PIF enhancement and a 1 : 1 stoichiometry was found for the β-CD and HP-β-CD complexes formed with TFV. The negative free energy (ΔG°) value indicated that the reaction of TFV with cyclodextrins was thermodynamically favorable. Furthermore, the structures of inclusion complexes of TFV with cyclodextrins were elucidated by 3-21G ab initio calculations. The limits of detection and quantification obtained ranged between 1.3 and 4.0 ng mL-1 and from 4.4 to 13.0 ng mL-1 in β-CD and HP-β-CD media, respectively. The analytical application in tap and river water samples yielded satisfactory mean recoveries ranging from 98.12 to 102.97%. Due to its sensitivity and ease of use, this method can be reliably applied to routine analysis.
{"title":"Cyclodextrin-enhanced photo-induced fluorescence of <i>tau</i>-fluvalinate, molecular modelling of inclusion complexes and determination in natural waters.","authors":"El Hadji Tombé Bodian, Coumba Faye, Diène Diégane Thiaré, Ndeye Arame Diop, Pape Abdoulaye Diaw, François Delattre, Atanasse Coly, Philippe Giamarchi","doi":"10.1039/d4ay00326h","DOIUrl":"10.1039/d4ay00326h","url":null,"abstract":"<p><p>The effect of adding organized supramolecular systems such as β-cyclodextrin (β-CD) and hydroxypropyl-β-cyclodextrin (HP-β-CD) on the photochemically-induced fluorescence (PIF) spectral properties of <i>tau</i>-fluvalinate (TFV) in aqueous solutions was examined. The influence of pH, UV irradiation time and photoproduct stability on the cyclodextrin-enhanced photochemically-induced fluorescence intensity was also investigated. The spectral changes associated with the inclusion process yielded values for the formation constants of TFV inclusion complexes between 450 and 640 M<sup>-1</sup>, which were calculated using the nonlinear iterative regression approach least squares. In addition, host-guest interaction was clearly determined by PIF enhancement and a 1 : 1 stoichiometry was found for the β-CD and HP-β-CD complexes formed with TFV. The negative free energy (Δ<i>G</i>°) value indicated that the reaction of TFV with cyclodextrins was thermodynamically favorable. Furthermore, the structures of inclusion complexes of TFV with cyclodextrins were elucidated by 3-21G <i>ab initio</i> calculations. The limits of detection and quantification obtained ranged between 1.3 and 4.0 ng mL<sup>-1</sup> and from 4.4 to 13.0 ng mL<sup>-1</sup> in β-CD and HP-β-CD media, respectively. The analytical application in tap and river water samples yielded satisfactory mean recoveries ranging from 98.12 to 102.97%. Due to its sensitivity and ease of use, this method can be reliably applied to routine analysis.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141416694","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sensitive and accurate determination of glyphosate (GLYP) is vital for food safety and environmental protection. Herein, a novel electrochemical ratiometric biosensor was designed for the accurate quantification of GLYP through one-step electrodeposition of MWCNTs-Cu MOF films. MWCNTs-Cu MOF nanostructures were directly electro-synthesized in situ on the electrode from the precursor solution. The combination of Cu MOFs with MWCNTs not merely improved the conductivity of MOFs, but also enhanced the sensitivity of the biosensor. Furthermore, Cu sites within Cu MOFs were turned into CuCl to further amplify the current signal and enable the specific recognition of GLYP through competing reactions with the transformation of CuCl into non-electroactive Cu-GLYP. Meanwhile, internal reference molecules of methylene blue (MB) were incorporated to improve the measurement accuracy of GLYP for reducing unpredictable measurement errors aroused by environmental deviations. The ratiometric electrochemical sensor exhibited a high linearity with the logarithmic value of GLYP concentration from 0.5 nM to 400 nM. The detection limit was estimated to be as low as 0.014 nM. Finally, the present sensor with ratiometric signal export was applied for GLYP analysis in real samples with high sensitivity and accuracy. The simplicity and reliability of the ratiometric sensor make it a worthy and powerful tool for food and environmental monitoring. This design strategy also provides an avenue for the development of simple and efficient biosensors for other substances.
{"title":"One-step electrodeposition of MWCNTs-Cu MOF films for the ratiometric electrochemical analysis of glyphosate.","authors":"Fan Zhao, Dongqing Guo, Jingyue Lan, Yunxi Liu","doi":"10.1039/d4ay00691g","DOIUrl":"10.1039/d4ay00691g","url":null,"abstract":"<p><p>Sensitive and accurate determination of glyphosate (GLYP) is vital for food safety and environmental protection. Herein, a novel electrochemical ratiometric biosensor was designed for the accurate quantification of GLYP through one-step electrodeposition of MWCNTs-Cu MOF films. MWCNTs-Cu MOF nanostructures were directly electro-synthesized <i>in situ</i> on the electrode from the precursor solution. The combination of Cu MOFs with MWCNTs not merely improved the conductivity of MOFs, but also enhanced the sensitivity of the biosensor. Furthermore, Cu sites within Cu MOFs were turned into CuCl to further amplify the current signal and enable the specific recognition of GLYP through competing reactions with the transformation of CuCl into non-electroactive Cu-GLYP. Meanwhile, internal reference molecules of methylene blue (MB) were incorporated to improve the measurement accuracy of GLYP for reducing unpredictable measurement errors aroused by environmental deviations. The ratiometric electrochemical sensor exhibited a high linearity with the logarithmic value of GLYP concentration from 0.5 nM to 400 nM. The detection limit was estimated to be as low as 0.014 nM. Finally, the present sensor with ratiometric signal export was applied for GLYP analysis in real samples with high sensitivity and accuracy. The simplicity and reliability of the ratiometric sensor make it a worthy and powerful tool for food and environmental monitoring. This design strategy also provides an avenue for the development of simple and efficient biosensors for other substances.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141425721","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bijay Kafle, Sileshi Gizachew Wubshet, Kari Anne Hestnes Bakke, Ulrike Böcker, Marion O'Farrell, Katinka Dankel, Ingrid Måge, Jon Tschudi, Dimitrios Tzimorotas, Nils Kristian Afseth, Tim Dunker
The main objective of this study was to design, build, and test a compact, multi-well, portable dry film FTIR system for industrial food and bioprocess applications. The system features dry film sampling on a circular rotating disc comprising 31 wells, a design that was chosen to simplify potential automation and robotic sample handling at a later stage. Calibration models for average molecular weight (AMW, 200 samples) and collagen content (68 samples) were developed from the measurements of industrially produced protein hydrolysate samples in a controlled laboratory environment. Similarly, calibration models for the prediction of lactate content in samples from cultivation media (59 samples) were also developed. The portable dry film FTIR system showed reliable model characteristics which were benchmarked with a benchtop FTIR system. Subsequently, the portable dry film FTIR system was deployed in a bioprocessing plant, and protein hydrolysate samples were measured at-line in an industrial environment. This industrial testing involved building a calibration model for predicting AMW using 60 protein hydrolysate samples measured at-line using the portable dry film FTIR system and subsequent model validation using a test set of 26 samples. The industrial calibration in terms of coefficient of determination (R2 = 0.94), root mean square of cross-validation (RMSECV = 194 g mol-1), and root mean square of prediction (RMSEP = 162 g mol-1) demonstrated low prediction errors as compared to benchtop FTIR measurements, with no statistical difference between the calibration models of the two FTIR systems. This is to the authors' knowledge the first study for developing and employing a portable dry film FTIR system in the enzymatic protein hydrolysis industry for successful at-line measurements of protein hydrolysate samples. The study therefore suggests that the portable dry film FTIR instrument has huge potential for in/at-line applications in the food and bioprocessing industries.
{"title":"A portable dry film FTIR instrument for industrial food and bioprocess applications.","authors":"Bijay Kafle, Sileshi Gizachew Wubshet, Kari Anne Hestnes Bakke, Ulrike Böcker, Marion O'Farrell, Katinka Dankel, Ingrid Måge, Jon Tschudi, Dimitrios Tzimorotas, Nils Kristian Afseth, Tim Dunker","doi":"10.1039/d4ay00238e","DOIUrl":"10.1039/d4ay00238e","url":null,"abstract":"<p><p>The main objective of this study was to design, build, and test a compact, multi-well, portable dry film FTIR system for industrial food and bioprocess applications. The system features dry film sampling on a circular rotating disc comprising 31 wells, a design that was chosen to simplify potential automation and robotic sample handling at a later stage. Calibration models for average molecular weight (AMW, 200 samples) and collagen content (68 samples) were developed from the measurements of industrially produced protein hydrolysate samples in a controlled laboratory environment. Similarly, calibration models for the prediction of lactate content in samples from cultivation media (59 samples) were also developed. The portable dry film FTIR system showed reliable model characteristics which were benchmarked with a benchtop FTIR system. Subsequently, the portable dry film FTIR system was deployed in a bioprocessing plant, and protein hydrolysate samples were measured at-line in an industrial environment. This industrial testing involved building a calibration model for predicting AMW using 60 protein hydrolysate samples measured at-line using the portable dry film FTIR system and subsequent model validation using a test set of 26 samples. The industrial calibration in terms of coefficient of determination (<i>R</i><sup>2</sup> = 0.94), root mean square of cross-validation (RMSECV = 194 g mol<sup>-1</sup>), and root mean square of prediction (RMSEP = 162 g mol<sup>-1</sup>) demonstrated low prediction errors as compared to benchtop FTIR measurements, with no statistical difference between the calibration models of the two FTIR systems. This is to the authors' knowledge the first study for developing and employing a portable dry film FTIR system in the enzymatic protein hydrolysis industry for successful at-line measurements of protein hydrolysate samples. The study therefore suggests that the portable dry film FTIR instrument has huge potential for in/at-line applications in the food and bioprocessing industries.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141416693","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Renato Soares de Oliveira Lins, Anandhakumar Sukeri, Mauro Bertotti
A nanoporous gold microelectrode (NPG-μE) was fabricated and used for Pb(II) detection in seawater samples via square wave anodic stripping voltammetry (SWASV). The Au microelectrode (Au-μE) was fabricated by embedding a gold microfiber into a Pasteur pipette, and its surface was further modified by an anodization-electrochemical reduction (A-ECR) method, yielding the NPG-μE. The fabricated electrodes were characterized by cyclic voltammetry (CV) and field emission scanning electron microscopy (FE-SEM) for electrochemical and structural morphological investigations. SWASV results show a Pb(II) stripping peak at around -0.05 V vs. Ag/AgCl, sat. KCl, which is unusual for common Pb(II) detection (typically occurring at around -0.40 V) in anodic stripping voltammetry (ASV) analysis. The Pb(II) detection at less negative stripping potential is more beneficial. Hence, it exhibited anti-interference properties with Cd(II), which is attributed to the preferential deposition and stripping of the target analyte on the low-indexed crystal planes of the NPG structure. The calibration plot obtained by SWASV was linear in the concentration range of 0.1-10 μM, and the detection limit was found to be 57 nM (correlation coefficient of 0.9974). The NPG microsensor presented a 15-fold enhanced current response compared to Au-μE, with excellent sensitivity (27.2 μA μM-1 cm-2). The application of the NPG microsensor was examined by detecting Pb(II) in seawater samples, and a satisfactory performance was obtained.
{"title":"A home-made nanoporous gold microsensor for lead(II) detection in seawater with high sensitivity and anti-interference properties.","authors":"Renato Soares de Oliveira Lins, Anandhakumar Sukeri, Mauro Bertotti","doi":"10.1039/d4ay00698d","DOIUrl":"10.1039/d4ay00698d","url":null,"abstract":"<p><p>A nanoporous gold microelectrode (NPG-μE) was fabricated and used for Pb(II) detection in seawater samples <i>via</i> square wave anodic stripping voltammetry (SWASV). The Au microelectrode (Au-μE) was fabricated by embedding a gold microfiber into a Pasteur pipette, and its surface was further modified by an anodization-electrochemical reduction (A-ECR) method, yielding the NPG-μE. The fabricated electrodes were characterized by cyclic voltammetry (CV) and field emission scanning electron microscopy (FE-SEM) for electrochemical and structural morphological investigations. SWASV results show a Pb(II) stripping peak at around -0.05 V <i>vs.</i> Ag/AgCl, sat. KCl, which is unusual for common Pb(II) detection (typically occurring at around -0.40 V) in anodic stripping voltammetry (ASV) analysis. The Pb(II) detection at less negative stripping potential is more beneficial. Hence, it exhibited anti-interference properties with Cd(II), which is attributed to the preferential deposition and stripping of the target analyte on the low-indexed crystal planes of the NPG structure. The calibration plot obtained by SWASV was linear in the concentration range of 0.1-10 μM, and the detection limit was found to be 57 nM (correlation coefficient of 0.9974). The NPG microsensor presented a 15-fold enhanced current response compared to Au-μE, with excellent sensitivity (27.2 μA μM<sup>-1</sup> cm<sup>-2</sup>). The application of the NPG microsensor was examined by detecting Pb(II) in seawater samples, and a satisfactory performance was obtained.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141430880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The analytical determination of opiates in biological samples is a critical mission and remains a challenge for almost all judicial and clinical drug testing panels due to their high abuse potential. Based on the high sensitivity of the longitudinal surface plasmon resonance (LSPR) peak of gold nanorods (AuNRs), we successfully developed a novel and simple refractive index sensing platform for detection of morphine (MOR) and codeine (COD) by means of 2-amino-5-mercapto-1,3,4-thiadiazole functionalized gold nanorods (AMTD-AuNRs) in aqueous solution, which is, to the best of our knowledge, the first report on the assay of MOR and COD using AuNRs. AMTD molecules strongly anchor onto the tips of AuNRs via the mercapto group and subsequent hydrogen-bonding interactions between AMTD and the analytes induced end-to-end chain assembly of AuNRs and a consequent decrease of the LSPR absorption band at 850 nm along with a bathochromic shift and emergence of a new hybridized plasmon mode at 1050 nm which was characterized using a Vis-NIR spectrophotometer. After systematic optimization, the absorbance ratio (A1050/A850) was proportional to the concentration of MOR in the ranges of 0.08-5 μM and 0.2-8 μM for COD without any significant effect from possible interferents. Furthermore, detection limits of 40 and 62 nM were achieved for MOR and COD, respectively, which are much lower than the cut-off level of 2000 ng mL-1 for opiates in urine samples set by the Substance and Abuse Mental Health Services Administration (SAMHSA). Eventually, as proof-of-applicability, human urine and blood serum samples spiked with MOR and COD were analyzed and excellent recoveries ranging from 94.4 to 108.9% were obtained, demonstrating the successful applicability of the designed refractive index probe in real biological specimens.
{"title":"A near-infrared plasmonic biosensor for detection of morphine and codeine in biological samples based on the end-to-end assembly of modified gold nanorods.","authors":"Naimeh Mohseni, Morteza Bahram","doi":"10.1039/d4ay00442f","DOIUrl":"10.1039/d4ay00442f","url":null,"abstract":"<p><p>The analytical determination of opiates in biological samples is a critical mission and remains a challenge for almost all judicial and clinical drug testing panels due to their high abuse potential. Based on the high sensitivity of the longitudinal surface plasmon resonance (LSPR) peak of gold nanorods (AuNRs), we successfully developed a novel and simple refractive index sensing platform for detection of morphine (MOR) and codeine (COD) by means of 2-amino-5-mercapto-1,3,4-thiadiazole functionalized gold nanorods (AMTD-AuNRs) in aqueous solution, which is, to the best of our knowledge, the first report on the assay of MOR and COD using AuNRs. AMTD molecules strongly anchor onto the tips of AuNRs <i>via</i> the mercapto group and subsequent hydrogen-bonding interactions between AMTD and the analytes induced end-to-end chain assembly of AuNRs and a consequent decrease of the LSPR absorption band at 850 nm along with a bathochromic shift and emergence of a new hybridized plasmon mode at 1050 nm which was characterized using a Vis-NIR spectrophotometer. After systematic optimization, the absorbance ratio (<i>A</i><sub>1050</sub>/<i>A</i><sub>850</sub>) was proportional to the concentration of MOR in the ranges of 0.08-5 μM and 0.2-8 μM for COD without any significant effect from possible interferents. Furthermore, detection limits of 40 and 62 nM were achieved for MOR and COD, respectively, which are much lower than the cut-off level of 2000 ng mL<sup>-1</sup> for opiates in urine samples set by the Substance and Abuse Mental Health Services Administration (SAMHSA). Eventually, as proof-of-applicability, human urine and blood serum samples spiked with MOR and COD were analyzed and excellent recoveries ranging from 94.4 to 108.9% were obtained, demonstrating the successful applicability of the designed refractive index probe in real biological specimens.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141430881","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The detection of anions using carbon dots (CDs) has received less attention compared to cations. Therefore, the present study aimed to develop a fluorescence sensor based on carbon dots (CDs) capable of detecting S2- in real water samples. The CDs were successfully prepared from the residues of a traditional Chinese herb, Gardenia, which emitted green photoluminescence (PL) under ultraviolet light irradiation. The as-prepared CDs were quasi-spherical in shape and ranged in size from 10 to 30 nm. Different detailed analyses proved that the CDs had good morphology, various functional groups, high water solubility, great optical features, and excellent stability under diverse environmental conditions. The ion detection showed that only Ag+ had the strongest fluorescence quenching effect on the CDs, however, the addition of S2- could recover their fluorescence. Based on these results, an "off-on" fluorescence sensor was achieved to selectively detect the concentration of S2- in real water samples with a limit of detection (LOD) of 39 μM, which further expanded the application of residues from traditional Chinese herbal medicine.
{"title":"Detection of sulphur(II) of carbon dots synthesized from <i>Gardenia</i> residue.","authors":"Zhaoxia Li, Yuchuan Dong, Xinyi Li, Dongchun Li, Jia Dong, Panchen Wang, Shuwei Chen, Huiling Geng","doi":"10.1039/d4ay00909f","DOIUrl":"10.1039/d4ay00909f","url":null,"abstract":"<p><p>The detection of anions using carbon dots (CDs) has received less attention compared to cations. Therefore, the present study aimed to develop a fluorescence sensor based on carbon dots (CDs) capable of detecting S<sup>2-</sup> in real water samples. The CDs were successfully prepared from the residues of a traditional Chinese herb, <i>Gardenia</i>, which emitted green photoluminescence (PL) under ultraviolet light irradiation. The as-prepared CDs were quasi-spherical in shape and ranged in size from 10 to 30 nm. Different detailed analyses proved that the CDs had good morphology, various functional groups, high water solubility, great optical features, and excellent stability under diverse environmental conditions. The ion detection showed that only Ag<sup>+</sup> had the strongest fluorescence quenching effect on the CDs, however, the addition of S<sup>2-</sup> could recover their fluorescence. Based on these results, an \"off-on\" fluorescence sensor was achieved to selectively detect the concentration of S<sup>2-</sup> in real water samples with a limit of detection (LOD) of 39 μM, which further expanded the application of residues from traditional Chinese herbal medicine.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141430882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Eugenia E Bashmakova, Alexander N Kudryavtsev, Alexey E Tupikin, Marsel R Kabilov, Aleksey E Sokolov, Ludmila A Frank
Melanoma inhibitory activity protein (MIA) does obviously offer the potential to reveal clinical manifestations of melanoma. Despite a pressing need for effective diagnosis of this highly fatal disease, there are no clinically approved MIA detection ELISA kits available. A recommended MIA threshold has not yet been defined, mostly by reason of variability in immunoglobulins' affinity and stability, the difference in sample preparation and assay conditions. Here we present a pair of high-affinity DNA aptamers developed as an alternative recognition and binding element for MIA detection. Their stability and reproducible synthesis are expected to ensure this analysis under standard conditions. The devised aptamer-based solid-phase microassay of model standard and control human sera involves luciferase NLuc as a highly sensitive reporter. Bioluminescence dependence on MIA concentration ranges in a linear manner from 2.5 to 250 ng mL-1, providing a MIA detection limit of 1.67 ± 0.57 ng mL-1.
黑色素瘤抑制活性蛋白(MIA)显然具有揭示黑色素瘤临床表现的潜力。尽管迫切需要对这种高度致命的疾病进行有效诊断,但目前还没有临床认可的 MIA 检测 ELISA 试剂盒。主要由于免疫球蛋白亲和力和稳定性的变化、样品制备和检测条件的差异,目前尚未确定推荐的 MIA 临界值。在此,我们提出了一对高亲和力 DNA 对映体,作为 MIA 检测的另一种识别和结合元件。它们的稳定性和可重复合成有望确保在标准条件下进行分析。所设计的基于适配体的固相微量检测模型标准和对照人类血清,采用了荧光素酶 NLuc 作为高灵敏度的报告物。生物发光对 MIA 浓度的依赖性在 2.5 至 250 ng mL-1 之间呈线性关系,MIA 检测限为 1.67 ± 0.57 ng mL-1。
{"title":"Bioluminescent aptamer-based microassay for detection of melanoma inhibitory activity protein (MIA).","authors":"Eugenia E Bashmakova, Alexander N Kudryavtsev, Alexey E Tupikin, Marsel R Kabilov, Aleksey E Sokolov, Ludmila A Frank","doi":"10.1039/d4ay00706a","DOIUrl":"10.1039/d4ay00706a","url":null,"abstract":"<p><p>Melanoma inhibitory activity protein (MIA) does obviously offer the potential to reveal clinical manifestations of melanoma. Despite a pressing need for effective diagnosis of this highly fatal disease, there are no clinically approved MIA detection ELISA kits available. A recommended MIA threshold has not yet been defined, mostly by reason of variability in immunoglobulins' affinity and stability, the difference in sample preparation and assay conditions. Here we present a pair of high-affinity DNA aptamers developed as an alternative recognition and binding element for MIA detection. Their stability and reproducible synthesis are expected to ensure this analysis under standard conditions. The devised aptamer-based solid-phase microassay of model standard and control human sera involves luciferase NLuc as a highly sensitive reporter. Bioluminescence dependence on MIA concentration ranges in a linear manner from 2.5 to 250 ng mL<sup>-1</sup>, providing a MIA detection limit of 1.67 ± 0.57 ng mL<sup>-1</sup>.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141425719","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Namita Kumari, Madhumati S Vaishnav, S S SRIKANTA, P. R. Krishnaswamy, Navakanta Bhat
Glycation and the subsequent formation of advanced glycation end products (AGEs) disrupt and impair the physiological functions of proteins. This study presents a comprehensive glycation site mapping of human serum albumin (HSA) utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS). Both in-vitro glycation experiments and patient samples were investigated, exploring various enzymes, processing techniques, and their impacts on glycation site detection. A pilot study was conducted, analyzing sixteen serum samples, which spanned from healthy individuals to severe diabetic patients (with HbA1c values ranging from 5.7% to 18.1%). The aim was to comprehend the progression of glycation on various sites of HSA with increasing levels of glycation. Their glycated albumin levels (GA) spanned from 19.7% to 62.3%. Trypsin-mediated proteolytic digestion unveiled 12 glycation sites through direct in-solution digestion of whole serum. However, isolating albumin from serum enabled the identification of a higher number of glycation sites in each sample compared to direct serum digestion. Boronate affinity chromatography facilitated the segregation of less glycated albumin (LGA) from the more glycated albumin (MGA) fraction. Subsequent proteolytic digestion of both LGA and MGA samples revealed similar glycation sites. The MGA fraction exhibited a greater number of identified glycation sites, thereby elucidating which sites are particularly prone to glycation in highly glycated albumin samples. Changes in relative glycation levels were noted in the tryptic digests of albumin samples following the sample enrichment steps, as opposed to direct in-solution digestion of whole serum. Two enzymes, trypsin and Glu-C, were evaluated for efficacy in sequence coverage and glycation site analysis of HSA, with trypsin demonstrating superior efficiency over Glu-C.
{"title":"Exploring Glycated Sites in Human Serum Albumin: Impact of Sample Processing Techniques on Detection and Analysis","authors":"Namita Kumari, Madhumati S Vaishnav, S S SRIKANTA, P. R. Krishnaswamy, Navakanta Bhat","doi":"10.1039/d4ay00503a","DOIUrl":"https://doi.org/10.1039/d4ay00503a","url":null,"abstract":"Glycation and the subsequent formation of advanced glycation end products (AGEs) disrupt and impair the physiological functions of proteins. This study presents a comprehensive glycation site mapping of human serum albumin (HSA) utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS). Both in-vitro glycation experiments and patient samples were investigated, exploring various enzymes, processing techniques, and their impacts on glycation site detection. A pilot study was conducted, analyzing sixteen serum samples, which spanned from healthy individuals to severe diabetic patients (with HbA1c values ranging from 5.7% to 18.1%). The aim was to comprehend the progression of glycation on various sites of HSA with increasing levels of glycation. Their glycated albumin levels (GA) spanned from 19.7% to 62.3%. Trypsin-mediated proteolytic digestion unveiled 12 glycation sites through direct in-solution digestion of whole serum. However, isolating albumin from serum enabled the identification of a higher number of glycation sites in each sample compared to direct serum digestion. Boronate affinity chromatography facilitated the segregation of less glycated albumin (LGA) from the more glycated albumin (MGA) fraction. Subsequent proteolytic digestion of both LGA and MGA samples revealed similar glycation sites. The MGA fraction exhibited a greater number of identified glycation sites, thereby elucidating which sites are particularly prone to glycation in highly glycated albumin samples. Changes in relative glycation levels were noted in the tryptic digests of albumin samples following the sample enrichment steps, as opposed to direct in-solution digestion of whole serum. Two enzymes, trypsin and Glu-C, were evaluated for efficacy in sequence coverage and glycation site analysis of HSA, with trypsin demonstrating superior efficiency over Glu-C.","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141515605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Food and Drug Administration (FDA) recently reported a new nitrosamine impurity in sitagliptin that was named as Nitroso-STG-19 (NTTP), whose acceptable intake limit was extremely low at 37 ng per day. Besides, NTTP was found to be a degradation impurity in sitagliptin tablet, which was formed by the reaction of 3-(Trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-A]pyrazine hydrochloride and nitrite salts introduced via excipients. Consequently, NTTP content in tablets was larger than that in API. To control the impurity, an ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) procedure for the detection of NTTP in both sitagliptin phosphate tablet and active pharmaceutical ingredient (API) was developed and validated. Furthermore, a derivatization method for the detection of nitrite salts at lower concentration was developed to select applicable excipients to decelerate the generation of NTTP. During validation of the analytical procedure for NTTP, quantitation limit (LOQ) of NTTP was 56 ppb (0.056 ng/mL), linear correlation coefficient was 0.9998, and recoveries of NTTP in spiked samples ranged from 95.5% to 105.2%, indicating the method is rapid, sensitive and accurate for NTTP test. In the nitrite salts detection method, LOQ was 0.21 ng/mL, recoveries of NTTP in spiked samples ranged from 87.6 % to 107.8%, indicating a sensitive and accurate method and it was suitable for screening appropriate pharmaceutical excipients.
{"title":"Exploration and Detection of Nitrosamine Impurity Nitroso-STG-19 in Sitagliptin Tablet and API as well as Nitrites in Excipients by LC-MS/MS Methods","authors":"Yajie Hao, Juan Fu, Ruixia Wei, Hao Teng, Guang Yin, Qihui Cao, Zhong Feng, Guimin Zhang","doi":"10.1039/d4ay00967c","DOIUrl":"https://doi.org/10.1039/d4ay00967c","url":null,"abstract":"Food and Drug Administration (FDA) recently reported a new nitrosamine impurity in sitagliptin that was named as Nitroso-STG-19 (NTTP), whose acceptable intake limit was extremely low at 37 ng per day. Besides, NTTP was found to be a degradation impurity in sitagliptin tablet, which was formed by the reaction of 3-(Trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-A]pyrazine hydrochloride and nitrite salts introduced via excipients. Consequently, NTTP content in tablets was larger than that in API. To control the impurity, an ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) procedure for the detection of NTTP in both sitagliptin phosphate tablet and active pharmaceutical ingredient (API) was developed and validated. Furthermore, a derivatization method for the detection of nitrite salts at lower concentration was developed to select applicable excipients to decelerate the generation of NTTP. During validation of the analytical procedure for NTTP, quantitation limit (LOQ) of NTTP was 56 ppb (0.056 ng/mL), linear correlation coefficient was 0.9998, and recoveries of NTTP in spiked samples ranged from 95.5% to 105.2%, indicating the method is rapid, sensitive and accurate for NTTP test. In the nitrite salts detection method, LOQ was 0.21 ng/mL, recoveries of NTTP in spiked samples ranged from 87.6 % to 107.8%, indicating a sensitive and accurate method and it was suitable for screening appropriate pharmaceutical excipients.","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141515608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Baijiang Jin, Gaojian Yang, Zhukang Guo, Zhu Chen, Yuan Liu, Song Li, Hui Chen, Yile Fang, Yan Deng, Nongyue He
Esophageal cancer is a common cancer with high morbidity and mortality that severely threatens the safety and quality of human life. The strong metastatic nature of esophageal cancer enables it to metastasize more quickly and covertly, making it difficult for current diagnostic and treatment methods to achieve efficient early screening, as well as timely and effective treatment. As a promising solution, nucleic acid aptamers, a kind of special single-stranded DNA or RNA oligonucleotide selected by the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) technology, can specifically bind with different molecular targets. In this paper, random DNA single-stranded oligonucleotides were used as the initial library. Using TE-1 cells and HEEC cells as targets, specific binding sequences were selected by 15 rounds of the cell-SELEX method, and the aptamer sequence that binds to TE-1 cells with the most specificity was obtained and named Te4. The Te4 aptamer was further validated for binding specificity, binding affinity, type of target, in vitro cytotoxicity when conjugated with DOX(Te4-DOX), and in vivo distribution. Results of in vitro validation showed that Te4 has outstanding binding specificity with a Kd value of 51.16 ± 5.52 nM, and the target type of Te4 was preliminarily identified as a membrane protein. Furthermore, the cytotoxicity experiment showed that Te4-DOX has specific cytotoxicity towards cultured TE-1 cells. Finally, the results of the in vivo distribution experiment showed that the Te4 aptamer is able to specifically target tumor regions in nude mice, showing great potential to be applied in future diagnosis and targeted therapy of esophageal cancer.
{"title":"Cell-SELEX and application research of a DNA aptamer against esophageal squamous cell carcinoma (ESCC) cell line TE-1.","authors":"Baijiang Jin, Gaojian Yang, Zhukang Guo, Zhu Chen, Yuan Liu, Song Li, Hui Chen, Yile Fang, Yan Deng, Nongyue He","doi":"10.1039/d4ay00895b","DOIUrl":"https://doi.org/10.1039/d4ay00895b","url":null,"abstract":"<p><p>Esophageal cancer is a common cancer with high morbidity and mortality that severely threatens the safety and quality of human life. The strong metastatic nature of esophageal cancer enables it to metastasize more quickly and covertly, making it difficult for current diagnostic and treatment methods to achieve efficient early screening, as well as timely and effective treatment. As a promising solution, nucleic acid aptamers, a kind of special single-stranded DNA or RNA oligonucleotide selected by the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) technology, can specifically bind with different molecular targets. In this paper, random DNA single-stranded oligonucleotides were used as the initial library. Using TE-1 cells and HEEC cells as targets, specific binding sequences were selected by 15 rounds of the cell-SELEX method, and the aptamer sequence that binds to TE-1 cells with the most specificity was obtained and named Te4. The Te4 aptamer was further validated for binding specificity, binding affinity, type of target, <i>in vitro</i> cytotoxicity when conjugated with DOX(Te4-DOX), and <i>in vivo</i> distribution. Results of <i>in vitro</i> validation showed that Te4 has outstanding binding specificity with a <i>K</i><sub>d</sub> value of 51.16 ± 5.52 nM, and the target type of Te4 was preliminarily identified as a membrane protein. Furthermore, the cytotoxicity experiment showed that Te4-DOX has specific cytotoxicity towards cultured TE-1 cells. Finally, the results of the <i>in vivo</i> distribution experiment showed that the Te4 aptamer is able to specifically target tumor regions in nude mice, showing great potential to be applied in future diagnosis and targeted therapy of esophageal cancer.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141490031","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}