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1.
Anal Chem ; 96(3): 1284-1292, 2024 01 23.
Artículo en Inglés | MEDLINE | ID: mdl-38194438

RESUMEN

In this work, a novel nanozyme (Cu@Zr) with all-in-one dual enzyme and fluorescence properties is designed by simple self-assembly. A nanozyme cascade sensor with disodium phenyl phosphate (PPDS) as substrate was first established by exploiting the dual enzymatic activities of phosphatase and laccase. Specifically, phosphatase cleaves the P-O bond of PPDS to produce colorless phenol, which is then oxidized by laccase and complexed with the chromogenic agent 4-aminoantipyrine (4-AP) to produce red quinoneimine (QI). Strikingly, the NH3 produced by the urease hydrolysis of urea can interact with Cu@Zr, accelerating the electron transfer rate and ultimately leading to a significantly improved performance of the cascade reaction. Moreover, the fluorescence at 440 nm of Cu@Zr is further quenched by the inner filter effect (IFE) of QI. Thus, the colorimetric and fluorescence dual-mode strategy for sensitive urease analysis with LODs of 3.56 and 1.83 U/L was established by the proposed cascade sensor. Notably, a portable swab loaded with Cu@Zr was also prepared for in situ urease detection with the aid of a smartphone RGB readout. It also provides a potentially viable analytical avenue for environmental and biological analysis.


Asunto(s)
Técnicas Biosensibles , Ureasa , Ureasa/química , Lacasa , Hidrólisis , Monoéster Fosfórico Hidrolasas , Colorimetría
2.
Mikrochim Acta ; 191(5): 282, 2024 04 23.
Artículo en Inglés | MEDLINE | ID: mdl-38652326

RESUMEN

A novel dual-mode fluorometric and colorimetric sensing platform is reported for determining glutathione S-transferase (GST) by utilizing polyethyleneimine-capped silver nanoclusters (PEI-AgNCs) and cobalt-manganese oxide nanosheets (CoMn-ONSs) with oxidase-like activity. Abundant active oxygen species (O2•-) can be produced through the CoMn-ONSs interacting with dissolved oxygen. Afterward, the pink oxDPD was generated through the oxidation of colorless N,N-diethyl-p-phenylenediamine (DPD) by O2•-, and two absorption peaks at 510 and 551 nm could be observed. Simultaneously, oxDPD could quench the fluorescence of PEI-AgNCs at 504 nm via the inner filter effect (IFE). However, in the presence of glutathione (GSH), GSH prevents the oxidation of DPD due to the reducibility of GSH, leading to the absorbance decrease at 510 and 551 nm. Furthermore, the fluorescence at 504 nm was restored due to the quenching effect of oxDPD on decreased PEI-AgNCs. Under the catalysis of GST, GSH and1-chloro-2,4-dinitrobenzo (CDNB) conjugate to generate an adduct, initiating the occurrence of the oxidation of the chromogenic substrate DPD, thereby inducing a distinct colorimetric response again and the significant quenching of PEI-AgNCs. The detection limits for GST determination were 0.04 and 0.21 U/L for fluorometric and colorimetric modes, respectively. The sensing platform illustrated reliable applicability in detecting GST in real samples.


Asunto(s)
Cobalto , Colorimetría , Glutatión Transferasa , Compuestos de Manganeso , Nanopartículas del Metal , Óxidos , Polietileneimina , Plata , Polietileneimina/química , Plata/química , Cobalto/química , Óxidos/química , Compuestos de Manganeso/química , Nanopartículas del Metal/química , Colorimetría/métodos , Glutatión Transferasa/metabolismo , Glutatión Transferasa/química , Límite de Detección , Oxidorreductasas/química , Oxidorreductasas/metabolismo , Humanos , Glutatión/química , Oxidación-Reducción , Técnicas Biosensibles/métodos , Fenilendiaminas/química , Nanoestructuras/química
3.
Mikrochim Acta ; 190(11): 444, 2023 10 18.
Artículo en Inglés | MEDLINE | ID: mdl-37851103

RESUMEN

A multi-signal aptasensor for thrombin determination is proposed based on catalytically active gold nanoparticles (AuNPs) and fluorescent silicon quantum dots (SiQDs). Yellow 4-Nitrophenol (4-NP) could be converted to colorless 4-Aminophenol (4-AP) by catalytically active aptamer-modified AuNPs (S1-AuNPs). The SiQDs emitted strong blue fluorescence at 455 nm at the excitation wavelength of 367 nm. When thrombin was absent, S1-AuNPs could catalytically reduce yellow 4-NP to colorless 4-AP. When thrombin was added, the aptamer could be transformed into a G-quadruplex structure, which masked the surface-active catalytic sites of AuNPs and restrained the reduction of 4-NP. Thus, the fluorescence of SiQDs was greatly quenched by 4-NP through the inner filter effect (IFE), and the solution color remained yellow. As the concentration of thrombin increased, the catalytic activity of S1-AuNPs decreased. The concentration of 4-NP that was converted to 4-AP declined and the unconverted 4-NP increased. In this process, the absorption peak of 4-NP at 400 nm increased while the fluorescence emission of SiQDs at 455 nm decreased. The linear ranges of the fluorometric and colorimetric aptasensor were 0.5-30 nM and 0.3-30 nM, respectively. The limits of detection (LOD) for the two modes were 0.15 nM and 0.13 nM. Furthermore, a portable sensing platform was constructed by combining the smartphone-based device with the software ImageJ for the determination of thrombin. With the advantages of cost-effectiveness, simplicity of operation and broad applicability, this aptasensor provided a new perspective for on-site determination of thrombin in the clinical field.


Asunto(s)
Aptámeros de Nucleótidos , Nanopartículas del Metal , Puntos Cuánticos , Puntos Cuánticos/química , Oro/química , Trombina , Silicio , Nanopartículas del Metal/química , Aptámeros de Nucleótidos/química , Colorantes
4.
Anal Chem ; 94(31): 11016-11022, 2022 08 09.
Artículo en Inglés | MEDLINE | ID: mdl-35899589

RESUMEN

Herein, based on electronic metal-support interaction (EMSI), a gold single atom confined MXene (AuSA/MXene) heterostructure was developed as the highly efficient electrochemiluminescence (ECL) functional material, which greatly improved the electrochemical properties and broadened the sensing application of MXenes. Gold single atoms were confined into the vacancy defects of Ti3C2Tx MXene, which could effectively avoid the masking of catalytic active sites. Meanwhile, electron transport could be accelerated with the assistance of titanium dioxide on the MXene nanosheets. Therefore, the AuSA/MXene heterostructure had high catalytic activity and electrical activity to promote hydrogen peroxide to generate free radicals, which achieved high-efficiency ECL. Eventually, the AuSA/MXene heterostructure was used to construct a Faraday cage-type ECL sensor with fluid nanoislands to detect miRNA-187 in triple-negative breast cancer tumor tissues.


Asunto(s)
Técnicas Electroquímicas , Oro , Oro/química , Mediciones Luminiscentes , Fotometría
5.
Analyst ; 147(4): 677-684, 2022 Feb 14.
Artículo en Inglés | MEDLINE | ID: mdl-35083988

RESUMEN

In this work, we report a novel and ultrasensitive dual-signal fluorescence emission detection system for protamine and trypsin based on the electrostatic interaction between polyethyleneimine (PEI) surface-modified positively charged carbon quantum dots (CDs-PEI) and the anionic fluorescent dye Eosin Y. The fluorescence system exhibited yellow-green fluorescence from Eosin Y and blue fluorescence from CDs-PEI. As a cationic peptide, protamine quenched the yellow-green fluorescence of Eosin Y at 542 nm through electrostatic interaction. In the presence of trypsin, protamine was specifically hydrolyzed by trypsin, which led to the subsequent recovery of the fluorescence of Eosin Y. Simultaneously, the blue fluorescence emission of CDs-PEI at 452 nm remained constant during the whole process. Hence, a ratiometric fluorescent nanoprobe for protamine and trypsin detection with high sensitivity was successfully constructed based on CDs-PEI and Eosin Y. For protamine detection, the ratiometric fluorescence intensity (I542/I452) exhibited an excellent linear relationship in the range of 0.1-5.2 µg mL-1 with a limit of detection (LOD) of 0.03 µg mL-1. And the linear relationship between I542/I452 and trypsin concentration ranged from 0.4 to 56 ng mL-1 with an LOD of 0.21 ng mL-1. Upon evaluating the performance of this method for the detection of trypsin in actual human urine samples, satisfactory results were finally obtained.


Asunto(s)
Polietileneimina , Puntos Cuánticos , Carbono , Eosina Amarillenta-(YS) , Colorantes Fluorescentes , Humanos , Límite de Detección , Protaminas , Espectrometría de Fluorescencia , Tripsina
6.
Anal Bioanal Chem ; 414(28): 7989-7998, 2022 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-36125540

RESUMEN

Herein, a simple and sensitive ratiometric fluorescence sensing platform to detect alkaline phosphatase (ALP) activity is developed on the basis of yellow fluorescent nitrogen-doped carbon quantum dots (YNCDs). The hydrolysis of ascorbic acid 2-phosphate (AAP) into ascorbic acid (AA) is catalyzed by ALP. Then, AA will react with o-phenylenediamine (OPD) to form 3-(1,2-dihydroxyethyl)furo[3,4b]-quinoxaline (QXD) which is a blue fluorescent quinoxaline derivative with emission at 435 nm in the presence of Cu2+. YNCDs have yellow fluorescence emission at 555 nm, and can maintain stable in QXD reaction system. Therefore, by utilizing the fluorescence of YNCDs at 555 nm as reference signal and the fluorescence of QXD at 435 nm as report signal, we can detect the ALP activity by monitoring the fluorescence ratio (F435/F555). The linear range is 0.5-5 U/L, and the limit of detection is 0.14 U/L. An application of this method for the analysis of ALP in human serum has given satisfactory results. A ratiometric fluorescent nanoprobe for ascorbic acid and alkaline phosphatase detection with excellent biocompatible and high sensitivity was successfully constructed based on YNCDs and QXD.


Asunto(s)
Puntos Cuánticos , Humanos , Fosfatasa Alcalina/análisis , Carbono , Nitrógeno , Fluorescencia , Espectrometría de Fluorescencia/métodos , Ácido Ascórbico , Quinoxalinas , Colorantes Fluorescentes , Límite de Detección
7.
Mikrochim Acta ; 189(4): 135, 2022 03 07.
Artículo en Inglés | MEDLINE | ID: mdl-35257215

RESUMEN

Iron-cobalt oxide nanosheets (FeCo-ONSs) were proved to have intrinsic peroxidase-like activity. Additionally, the peroxidase-like activity of FeCo-ONSs toward the oxidation of 3,3,5,5-tetramethylbenzidine (TMB) was dramatically enhanced after heparin addition due to the stronger affinity toward TMB. Protamine combines with heparin, so the promotion of peroxidase-like activity of FeCo-ONSs with heparin was suppressed. With the addition of trypsin, protamine was hydrolyzed and the enhancement effect of catalytic activity of FeCo-ONSs was recovered. Based on above process, a sensitive colorimetric platform for trypsin activity determination was constructed through measuring the absorbance of produced oxTMB at 652 nm, providing a linear detection range of 5 to 500 ng/mL and a low detection limit of 2.8 ng/mL. The method was applied to trypsin determination in real samples (human urine sample and multienzyme tablet sample) with satisfactory results, illustrating the potential application of this biosensor.


Asunto(s)
Colorimetría , Peroxidasa , Cobalto , Colorimetría/métodos , Heparina , Humanos , Hierro , Óxidos , Oxidorreductasas , Peroxidasas , Protaminas , Tripsina
8.
Mikrochim Acta ; 189(9): 363, 2022 09.
Artículo en Inglés | MEDLINE | ID: mdl-36044087

RESUMEN

A Co, N co-doped porous carbon-based nanozyme (Co-N-C nanozyme) has been fabricated. Taking advantages of the excellent oxidase catalytic activity and significant stability of Co-N-C nanozyme, we propose a fluorescence and colorimetric system based on Co-N-C nanozyme and red-emitting carbon quantum dots (RCDs) for butyrylcholinesterase (BChE) sensing. As the chromogenic substrate 3,3',5,5'-tetramethylbenzidine (TMB) was catalyzed and oxidized by Co-N-C nanozyme, the generated oxTMB had a new absorption peak at 652 nm, which resulted in the significant quenching of the fluorescence of the carbon quantum dots at 610 nm. Under the catalysis of BChE, thiocholine was generated from the hydrolysis of S-butyrylthiocholine iodide (BTCh), and the as-generated thiocholine effectively inhibited the oxidation of TMB catalyzed by Co-N-C nanozyme, leading to a decrease of the absorption of oxTMB at 652 nm and effective fluorescence recovery of RCDs. By measuring the absorbance of produced oxTMB at 652 nm and the fluorescence of RCDs at 610 nm, the fluorescence and colorimetric system both exhibited an outstanding linear response to the activity of BChE in the range 0.5 to 40 U L-1, with a detection limit of 0.16 U L-1 and 0.21 U L-1, respectively. Furthermore, this established dual-channel biosensing strategy has been successfully applied to the determination of BChE in human serum samples. The present work has effectively expanded the development and application of nanozyme in biosensing.


Asunto(s)
Técnicas Biosensibles , Butirilcolinesterasa , Colorimetría , Técnicas Biosensibles/métodos , Butirilcolinesterasa/análisis , Butirilcolinesterasa/química , Carbono , Colorimetría/métodos , Humanos , Nanoestructuras/química , Oxidorreductasas , Porosidad , Tiocolina
9.
Anal Chem ; 93(51): 17086-17093, 2021 12 28.
Artículo en Inglés | MEDLINE | ID: mdl-34914874

RESUMEN

MXene material has been gradually studied in recent years due to its fascinating characteristics. This work developed a novel MXene-derived quantum dots (MQDs) @gold nanobones (Au NBs) heterostructure as the electrochemiluminescence (ECL) sensor. First, MXene and MQDs were synthesized via the green preparation process, which avoided the harm of hydrofluoric acid to humans and the environment. There was a strong ECL signal enhancement in the MQD@Au NBs heterostructure. On the one hand, Au NBs with surface plasmon resonance (SPR) effect acted as an "electronic regulator" that can transfer electrons to itself to control over-injection of electrons into the conduction band of MQDs. The luminous signal of MQDs can be efficiently generated and significantly amplified in the ECL sensing process. On the other hand, the work function of MQDs with excellent conductivity was relatively close to that of Au NBs in the heterostructure. So, ECL quenching caused by short-distance electron transfer between luminophore and Au nanomaterial has been effectively suppressed. The MQD@Au NBs heterostructure-based ECL sensing system was applied to determine miRNA-26a in the serum of patients with triple-negative breast cancer. It not only provides ideas for the green synthesis of MXene but also provides a guide for the application of MQD@Au NBs heterostructure in the field of ECL sensing.


Asunto(s)
Técnicas Biosensibles , Nanopartículas del Metal , MicroARNs , Puntos Cuánticos , Neoplasias de la Mama Triple Negativas , Combinación de Medicamentos , Durapatita , Técnicas Electroquímicas , Oro , Humanos , Mediciones Luminiscentes , Dióxido de Silicio , Neoplasias de la Mama Triple Negativas/diagnóstico
10.
Anal Chem ; 93(6): 3130-3137, 2021 02 16.
Artículo en Inglés | MEDLINE | ID: mdl-33535742

RESUMEN

Herein, we designed a new strategy for fabricating a renewable bioresource-derived N-doped hierarchical porous carbon-supported iron (Fe/NPC)-based oxidase mimic. The obtained results suggested that Fe/NPC possessed a large specific surface area (1144 m2/g) and pore volume (0.62 cm3/g) to afford extensive Fe-Nx active sites. Taking advantages of the remarkable oxidase-mimicking activity, outstanding stability, and reusability of Fe/NPC, a novel dual-channel biosensing system was strategically fabricated for sensitively determining acetylcholinesterase (AChE) through the integration of Fe/NPC and fluorescent silver nanoclusters (AgNCs) for the first time. The limits of detection for AChE can achieve as low as 0.0032 and 0.0073 U/L by the outputting fluorometric and colorimetric dual signals, respectively. Additionally, this dual-signal system was applied to analyze human erythrocyte AChE and its inhibitor with robust analytical performance. This work provides one sustainable and effective avenue to apply a bioresource for fabricating an Fe/NPC-based oxidase mimic with high catalytic performance and also gives new impetuses for developing novel biosensors by applying Fe/NPC-based enzyme mimics as substitutes for the natural enzyme.


Asunto(s)
Carbono , Hierro , Colorimetría , Humanos , Oxidorreductasas , Porosidad
11.
Anal Chem ; 93(20): 7508-7515, 2021 05 25.
Artículo en Inglés | MEDLINE | ID: mdl-33970617

RESUMEN

A smart electrochemiluminescent (ECL) sensor has been designed in this work. The sensing system consisted of Ag NPs-Ti3AlC2 nanosheets (Ag-TACS) as the self-luminous Faraday cage and biomimetic magnetic vesicles as the functional substrate. By engineering the structure and properties of Ti3AlC2 nanosheets to induce the Faraday-cage effect, the outer Helmholtz plane (OHP) was extended to contribute to ECL enhancement. Compared with the Faraday cage that further incorporated luminous materials, the self-luminous Faraday cage in the "direct label" model kept all the luminous materials on the OHP. Meanwhile, biomimetic magneticvesicles with highly efficient fluidity were used to improve the sensing efficiency and obtain a perfect Faraday-cage structure to enhance the ECL signals. The highest ECL enhancement (ca. 25 times) has been achieved by the synergistic effect of the Faraday cage and biomimetic magnetic vesicles. This sensing system was used to detect the wild-type K-ras gene in the colorectal tumor tissue. It provides not only an important guide for the novel ECL sensing concept but also a smart modulation system of the electromagnetic field.


Asunto(s)
Técnicas Biosensibles , Técnicas Electroquímicas , Biomimética , Límite de Detección , Mediciones Luminiscentes , Fenómenos Magnéticos
12.
Analyst ; 146(17): 5280-5286, 2021 Sep 07.
Artículo en Inglés | MEDLINE | ID: mdl-34342310

RESUMEN

Herein, we designed a diversified sensing platform for d-penicillamine based on amino-functionalized Zr-based metal-organic frameworks (UiO-66-NH2 MOFs) and 3-aminophenylboronic acid (APBA)@Alizarin Red (ARS). The boronic acid group of 3-aminophenylboronic acid could react with Alizarin Red to form an APBA@ARS complex with a yellow fluorescence emission at 580 nm and ultraviolet absorption at 435 nm. APBA@ARS can greatly quench the fluorescence of UiO-66-NH2 MOFs at 450 nm via fluorescence resonance energy transfer (FRET). When copper ions were present in the reaction system of APBA and Alizarin Red, the copper ions could complex with Alizarin Red to prevent the generation of APBA@ARS, and the absorption of Cu@ARS at 530 nm occurred. Thus, the absorbance of APBA@ARS at 435 nm declined, restoring the fluorescence of UiO-66-NH2 MOFs. Nevertheless, when d-penicillamine and copper ions coexist in the APBA and Alizarin Red reaction system, the copper ions would complex with the sulfhydryl group of d-penicillamine and no longer hinder the generation of APBA@ARS, and the fluorescence of UiO-66-NH2 MOFs is quenched again. Meanwhile, the absorbance of APBA@ARS at 435 nm enhanced and the absorbance at 530 nm decreased. Thus, a fluorescence and colorimetric dual-signal sensing platform was constructed for d-penicillamine detection, which could detect d-penicillamine in the 1-20 µM and 2-50 µM ranges with the limit of detection (LOD) values of 0.46 µM and 1.38 µM, respectively. Furthermore, this sensing platform could also realize the intelligent RGB detection via mobile phones due to the obvious color change of the reaction system.


Asunto(s)
Ácidos Borónicos , Penicilamina , Antraquinonas
13.
Analyst ; 146(3): 896-903, 2021 Feb 07.
Artículo en Inglés | MEDLINE | ID: mdl-33237050

RESUMEN

Single-atom nanozymes have drawn wide attention in bio-sensing for their remarkable merits such as low cost, high stability, and maximum atom utilization. Herein, a colorimetric strategy based on Fe-N-C single-atom nanozymes (Fe/NC-SAs) was established for the detection of alkaline phosphatase (ALP) activity. The Fe/NC-SAs prepared by pyrolysis have excellent peroxidase-like activity and can oxidize 3,3',5,5'-tetramethylbenzidine (TMB) to a blue color product in the presence of hydrogen peroxide (H2O2). When ascorbic acid (AA) is added to the system, the blue color fades, and the absorbance has a linear relationship with the concentration of AA. Alkaline phosphatase (ALP) can catalyze the hydrolysis of ascorbic acid 2-phosphate (AAP) to produce AA. Thus, a strategy based on Fe/NC-SAs for the detection of ALP activity was established, which provided a linear range of 0.1-1.5 U L-1 and a limit of detection as low as 0.05 U L-1. Besides, Fe/NC-SAs showed high stability under harsh conditions. Moreover, an Fe/NC-SA-based assay was successfully validated using human serum samples for ALP determination with satisfactory results, and has broad prospects in the field of biosensing.


Asunto(s)
Fosfatasa Alcalina , Peróxido de Hidrógeno , Fosfatasa Alcalina/metabolismo , Colorimetría , Humanos , Límite de Detección , Oxidación-Reducción , Peroxidasas
14.
Anal Bioanal Chem ; 413(23): 5779-5787, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34312692

RESUMEN

A simple and rapid ratiometric fluorescent sensing system for D-penicillamine (D-PA) determination is developed based on yellow carbon dots (Y-CDs) combined with thiochrome (oxVB1) for the first time. The oxidization of thiamine (VB1) can be catalyzed by Alkaline-hydrolyzed artemisinin (a-ART) to form oxVB1, which leads to the occurrence of fluorescence emission peak at 466 nm. Furthermore, the oxidation reaction between a-ART and VB1 could be inhibited by D-PA, and accompanied with the decrease of fluorescence at 466 nm. However, the fluorescence peak of Y-CDs as an internal reference at 566 nm was almost unchanged. The ratiometric signal changes contributed to a robust and sensitive D-PA sensing. Under the optimal condition, a good linear response for the D-PA detection was obtained in the ranges of 0.5-50 µM with a detection limit of 0.33 µM. In addition, Y-CDs and thiochrome-based sensing system was applied to D-PA determination in real samples and obtained acceptable results. We developed a new carbon dots/thiochrome fluorescent nanoprobe for ratiometric fluorescence sensing of D-penicillamine.


Asunto(s)
Carbono/química , Penicilamina/análisis , Puntos Cuánticos/química , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Tiamina/análogos & derivados , Catálisis , Humanos , Límite de Detección , Penicilamina/sangre , Tiamina/química
15.
Mikrochim Acta ; 188(7): 227, 2021 06 10.
Artículo en Inglés | MEDLINE | ID: mdl-34109464

RESUMEN

A sensitive fluorescence strategy was constructed for the detection of α-glucosidase activity based on AgInZnS QDs. The AIZS QDs which were synthesized by hydrothermal method have a fluorescence emission wavelength of 554 nm. Ce4+ was able to oxidize p-phenylenediamine (PPD) to generate oxPPD, which can quench the fluorescence of AIZS QDs through dynamic quenching. When α-glucosidase was introduced into the system, L-ascorbic acid-2-O-α-D-glucopyranosyl (AAG) could be hydrolyzed to form ascorbic acid (AA), which can reduce Ce4+ and prevent the oxidation of PPD. Thus, the dynamic quenching process was blocked accompanying with the fluorescence recovery of AIZS QDs. The developed detection system for α-glucosidase displayed a good linear relationship between 0.01 and 0.16 U·mL-1 with a detection limit of 0.0073 U·mL-1. The sensing platform with high feasibility and anti-interference is a competitive alternative applied to α-glucosidase-related diagnostics.


Asunto(s)
Pruebas de Enzimas/métodos , Fluorometría/métodos , Puntos Cuánticos/química , alfa-Glucosidasas/metabolismo , Humanos
16.
Analyst ; 145(3): 1001-1007, 2020 Feb 03.
Artículo en Inglés | MEDLINE | ID: mdl-31830153

RESUMEN

Herein, papain-protected bimetallic gold/silver nanoclusters (Au/Ag NCs) were successfully synthesized and applied for the detection of ascorbate oxidase (AAO). The doping of papain-protected Au nanoclusters with Ag enhanced the fluorescence intensity with an intense red fluorescence peak at 617 nm, and the red-emitting Au/Ag nanoclusters were further used to monitor the AAO activity. The fluorescence of Au/Ag NCs could be quenched by hydrogen peroxide (H2O2) due to the generation of hydroxyl radicals (˙OH) from the reaction of Ag/Au nanoclusters and H2O2. However, the addition of ascorbic acid (AA) effectively reacted with the free radicals and caused the fluorescence recovery of the Au/Ag NCs. Furthermore, AAO could catalyze the oxidation of AA to form dehydro-ascorbate (DHA). As a result, there was not enough AA to consume the hydroxyl radicals, which resulted in a decrease in the fluorescence of the papain-capped Au/Ag NCs. Therefore, the AAO activity can be monitored by measuring the fluorescence intensity of the red-emitting Au/Ag NCs. Moreover, the developed method for AAO detection displayed a good linear relationship from 5 to 80 mU mL-1 and the detection limit was 1.72 mU mL-1. Thus, a simple and selective method for the determination of the AAO activity was constructed and satisfactory results were obtained in real sample detection.


Asunto(s)
Ascorbato Oxidasa/metabolismo , Oro/química , Nanopartículas del Metal/química , Plata/química , Espectrometría de Fluorescencia/métodos , Ascorbato Oxidasa/sangre , Ácido Ascórbico/análisis , Ácido Ascórbico/química , Ácido Ascórbico/metabolismo , Biocatálisis , Humanos , Peróxido de Hidrógeno/química , Radical Hidroxilo/química , Radical Hidroxilo/metabolismo , Límite de Detección , Oxidación-Reducción , Papaína/química , Papaína/metabolismo , Reproducibilidad de los Resultados
17.
Analyst ; 145(17): 5808-5815, 2020 Aug 24.
Artículo en Inglés | MEDLINE | ID: mdl-32672281

RESUMEN

In this work, a novel ratiometric fluorescent platform for α-glucosidase (α-glu) and its inhibitor was constructed based on N-doped carbon dots (N-CDs). The α-glucosidase present can catalyze the release of hydroquinone (HQ) from α-arbutin. Then, the generated HQ can be oxidized and copolymerized with polyethyleneimine (PEI) to form a yellowish green fluorescence copolymer (PHQ-PEI) with intense fluorescence emission at 510 nm. When the PHQ-PEI was formed, blue fluorescence of N-CDs at 425 nm was decreased, whereas the fluorescence of PHQ-PEI at 510 nm increased sharply as a result of the fluorescence resonance energy transfer (FRET) effect between N-CDs and PHQ-PEI. However, in the presence of acarbose, the activity of α-glucosidase is inhibited, and α-arbutin cannot be hydrolyzed to hydroquinone, leading to the fluorescence recovery of N-CDs at 425 nm and the fluorescence decrease of PHQ-PEI at 510 nm. The linear range from 0.2 to 1.6 mU mL-1 and 25-150 µmol L-1 was obtained for α-glucosidase and acarbose detection, respectively, and the detection limit (LOD) for α-glucosidase and acarbose was as low as 0.082 mU mL-1 and 14.5 µmol L-1. Thus, a ratiometric fluorescent sensor with good sensitivity and high specificity was established for α-glucosidase assay and satisfactory results were acquired in real sample determination.


Asunto(s)
Técnicas Biosensibles , Puntos Cuánticos , Acarbosa , Carbono , alfa-Glucosidasas
18.
Mikrochim Acta ; 187(9): 532, 2020 08 31.
Artículo en Inglés | MEDLINE | ID: mdl-32864710

RESUMEN

A novel fluorescent sensing platform based on nitrogen-doped graphene quantum dots (N-GQDs) is presented, which is able to detect various metabolites (cholesterol, glucose, lactate, and xanthine) rapidly, sensitively, and selectively. Hg2+ can attach on the surface of N-GQDs, leading to the quenching of N-GQD fluorescence. In the presence of cysteine (Cys), Hg2+ is released from N-GQDs and associates with Cys. Then, the fluorescence of N-GQDs is recovered. Hydrogen peroxide, resulting from the enzymatic oxidation of metabolites, can convert two molecules of Cys into one molecule of cystine, which cannot bind with Hg2+. So, the fluorescence of N-GQDs quenched again. For cholesterol, glucose, lactate, and xanthine, the limits of detection are 0.035 µmol/L, 0.025 µmol/L, 0.07 µmol/L, and 0.04 µmol/L, respectively, and the linear ranges are 1-12 µmol/L, 0.06-3 µmol/L, 0.2-70 µmol/L, and 0.12-17 µmol/L, respectively. The presented method was applied to quantify metabolites in human blood samples with satisfactory results. Graphical abstract.


Asunto(s)
Glucemia/análisis , Colesterol/sangre , Ácido Láctico/sangre , Puntos Cuánticos/química , Espectrometría de Fluorescencia/métodos , Xantina/sangre , Cisteína/química , Grafito/química , Humanos , Peróxido de Hidrógeno/química , Límite de Detección , Mercurio/química , Nitrógeno/química , Oxidación-Reducción
19.
Analyst ; 144(24): 7398-7405, 2019 Dec 21.
Artículo en Inglés | MEDLINE | ID: mdl-31670357

RESUMEN

α-Glucosidase and its inhibitors play a key role in diagnosis and treatment of diabetes. In the present work, we established a facile, sensitive and selective fluorescence method based on silicon quantum dots (SiQDs) and MnO2 nanosheets for the determination of α-glucosidase and one of its inhibitors acarbose. The fluorescence of SiQDs was greatly quenched by MnO2 nanosheets due to the inner filter effect. α-Glucosidase could easily catalyze the hydrolysis of l-ascorbic acid-2-O-α-d-glucopyranosyl (AAG) to produce ascorbic acid (AA), which could reduce MnO2 nanosheets to Mn2+, resulting in dramatic recovery of the fluorescence of SiQDs. The proposed sensing platform could provide a good linear relationship between the fluorescence intensity of SiQDs and the concentration of α-glucosidase in the range of 0.02-2.5 U mL-1 with a detection limit of 0.007 U mL-1. In addition, the sensing platform could be used for α-glucosidase inhibition. Acarbose was one of the most common and typical inhibitors, and this sensing platform can be utilized to detect acarbose in the range of 1-1000 µM. The developed fluorescence method was successfully validated for the determination of α-glucosidase in human serum samples.


Asunto(s)
Colorantes Fluorescentes/química , Inhibidores de Glicósido Hidrolasas/análisis , Nanocompuestos/química , Puntos Cuánticos/química , alfa-Glucosidasas/sangre , Humanos , Compuestos de Manganeso/química , Óxidos/química , Silicio/química , Espectrometría de Fluorescencia
20.
Mikrochim Acta ; 186(12): 839, 2019 11 23.
Artículo en Inglés | MEDLINE | ID: mdl-31760490

RESUMEN

A fluorometric method is described for the detection of alkaline phosphatase (ALP) activity. It is based on the use of the product of hydrolysis of the drug amifostine (a thiophosphoester) by ALP. It is known that MnO2 nanosheets quench the blue fluorescence of tungsten disulfide quantum dots (WS2 QDs) which have excitation/emission wavelengths of 320/448 nm. However, in the presence of ALP and amifostine, the product of hydrolysis [2-(3-aminopropylamino)ethanethiol] triggers the decomposition of the MnO2 nanosheets. This results in the recovery of fluorescence. Based on this finding, an assay for ALP activity was developed that works in the 0.09-1.6 U L-1 range, with a 40 mU L-1 detection limit. The relative standard deviation is 1.87% for five repeated measurements of 0.8 U L-1 ALP. The method was applied to the analysis of ALP in real samples and gave satifactory results. Graphical abstractSchematic representation of a fluorometric method for determination of the activity of alkaline phosphatase (ALP). The fluorescence of a system composed of WS2 quantum dots and MnO2 nanosheets is quenched. Hydrolysis of the cytoprotective adjuvant amifostine (a phosphothioester) by ALP leads to a thiol that causes the decomposition of the MnO2 nanosheets. As a result, the blue fluorescence of the system becomes increasingly restored.


Asunto(s)
Fosfatasa Alcalina/análisis , Fluorometría , Compuestos de Manganeso/química , Nanopartículas/química , Óxidos/química , Puntos Cuánticos/química , Compuestos de Tungsteno/química , Fosfatasa Alcalina/metabolismo , Estructura Molecular , Tamaño de la Partícula , Propiedades de Superficie
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