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1.
Anal Chim Acta ; 1191: 339363, 2022 Jan 25.
Article in English | MEDLINE | ID: mdl-35033235

ABSTRACT

We present a novel dual-imprinted electrochemical paper-based analytical device (Di-ePAD) to simultaneously determine 8-hydroxy-2'-deoxyguanosine (8-OHdG) and 3-nitrotyrosine (3-NT) and assess oxidative and nitrative biomarkers in urine and plasma samples. The Di-ePAD was designed with hydrophobic barrier layers formed on filter paper to provide three-dimensional circular reservoirs and assembled electrodes. The molecularly imprinted polymer (MIP) was synthesized using a silica nanosphere decorated with silver nanoparticles (SiO2@AgNPs) as a core covered with dual-analyte imprinted sites on the polymer to recognize selectively and bind the target biomarkers. This strategy drives monodispersity and enhances the conductivity of the resulting MIP core-shell products. 3-NT-MIP and 8-OHdG-MIP were synthesized by successively coating the surface of SiO2@AgNPs with l-Cysteine via the thiol group, then terminating with MIP shells. The dual imprinted core-shell composites possess attractive properties for the target biomarkers' sensing, including catalytic activity, selectivity, and good conductivity. The Di-ePAD revealed excellent linear dynamic ranges of 0.01-500 µM for 3-NT and 0.05-500 µM for 8-OHdG, with detection limits of 0.0027 µM for 3-NT and 0.0138 µM for 8-OHdG. This newly developed method based on the synergistic effects of SiO2@AgNPs combined with promising properties of MIP offers outstanding selectivity, sensitivity, reproducibility, simplicity, and low cost for quantitative analysis of 3-NT and 8-OHdG. The proposed Di-ePAD showed good accuracy and precision when applied to actual samples, including urine and serum samples validated by a conventional HPLC method.


Subject(s)
Metal Nanoparticles , Molecular Imprinting , Biomarkers , Electrochemical Techniques , Electrodes , Limit of Detection , Oxidative Stress , Reproducibility of Results , Silicon Dioxide , Silver
2.
Anal Chim Acta ; 1191: 339290, 2022 Jan 25.
Article in English | MEDLINE | ID: mdl-35033237

ABSTRACT

In this study, we developed novel, simple gravimetric and voltammetric sensors for the ultrasensitive detection of active matrix metalloproteinase (MMP)-2 in plasma. The developed sensors are cost-effective, require a very less amount of reagents, and are time-saving. They detect MMP-2 based on antigen-antibody recognition and its ability to cleave glycine-leucine peptide bond. The three-dimensional bioplatform of the sensors consisted of a cationic polyethyleneimine (PEI) polymer that facilitated robust immobilization of the dipeptide labeled with anthraquinone (AQ), or antibody molecules in appropriate density, which was crucial for biosensing. Detection was performed using quartz crystal microbalance with dissipation and voltammetry. The results showed that the developed sensors were characterized by high stability, wide analytical range (2.0 pg mL-1 to 5.0 µg mL-1), and low detection limit (ca. 10 fg mL-1). They also exhibited excellent efficiency in the determination of active MMP-2 in real samples, such as blood plasma. The developed sensors may hold great promise for the early diagnosis of cancers.


Subject(s)
Biosensing Techniques , Matrix Metalloproteinase 2 , Biomarkers, Tumor , Electrochemical Techniques , Immunoassay , Limit of Detection , Plasma , Quartz Crystal Microbalance Techniques
3.
Anal Chim Acta ; 1191: 339314, 2022 Jan 25.
Article in English | MEDLINE | ID: mdl-35033249

ABSTRACT

In our research, label-free and surface-enhanced Raman dyes-free Raman spectroscopy which was used to detect carcinoembryonic antigen (CEA) according to poly adenine (Poly A)-regulated self-assembly methods was developed and studied. CEA induced partial hybridization of Ab-H2 and Ab-H1, and Ab-H1-CEA-Ab-H2 (a sandwich proximity CEA-DNA complex) was formed, which unfolded molecular beacon 1 (MB1) and modified the substrate. Subsequently, MB2-AuNPs were hybridized with MB1, and Ab-H1-CEA-Ab-H2 was released via toehold regulated displacements of DNA strands. Therefore, hybridization processes of MB2 and MB1 were induced and promoted by CEA-DNA complexes which worked as catalysts. The misplaced target then induced a next round of strand exchange, and the signals for determination of CEA were amplified by AuNPs absorbed on the substrate. It was indicated that the spectral characteristics of adenine at 736 cm-1 were consistent with the SERS spectrum of DNA. Adenine acted as an internal marker for label-free SERS detection of CEA. Moreover, satisfactory stability and reproducibility were found. Meanwhile, the antibody could specifically recognize the corresponding antigen. Since adenine was dominant in SERS spectra, which was also proximal to Au surface, the sensitivity of the novel method was high without modifications. The analytical performance of this method in determining serum CEA was satisfactory.


Subject(s)
Biosensing Techniques , Metal Nanoparticles , Carcinoembryonic Antigen , DNA , Gold , Limit of Detection , Reproducibility of Results , Spectrum Analysis, Raman
4.
Anal Chim Acta ; 1191: 339313, 2022 Jan 25.
Article in English | MEDLINE | ID: mdl-35033251

ABSTRACT

A rapid and highly sensitive vesicle mediated dispersive liquid-liquid microextraction procedure is developed for the determination of parts per quadrillion level of beryllium in seawater and air filter samples for providing its natural background and contamination levels. In this procedure, dioctylsulfosuccinate, an anionic vesicular surfactant and acetylacetone are used as dispersing and chelating agents, respectively. At pH > 9.5, beryllium forms hydrophobic beryllium-acetylacetonate complex spontaneously at room temperature. This complex is selectively filled into the vesicular cavities of dioctylsulfosuccinate and is extracted into small chloroform phase from bulk aqueous phase. The beryllium present in the chloroform phase is back extracted with dilute nitric acid and is analyzed by graphite furnace atomic absorption spectrometry. This method is applied to groundwater, seawater, coal fly ash, air filter and sea sludge samples. Under the optimized conditions, the limit of detection, limit of quantification and linear dynamic range are 10 fg mL-1, 33 fg mL-1 and 40-500 fg mL-1 for seawater; 0.15 ng g-1, 0.5 ng g-1 and 0.4-4 ng g-1 for air filter and 1.5 ng g-1, 0.39 ng g-1 and 0.4-4 ng g-1 for coal fly ash, respectively. For 1 L seawater sample an enrichment factor of 954 is achieved. Coefficient of determination (R2) is found to be 0.997. The recoveries are in the range of 94-105% at 200-500 fg mL-1. The relative standard deviations are 20%, 11%, 8% for ppq, ppt and ppb levels of Be, respectively. The accuracy of the procedure is verified by analyzing NIST SRMs 1640 and 1640a trace elements in natural water.


Subject(s)
Graphite , Liquid Phase Microextraction , Beryllium , Hydrogen-Ion Concentration , Limit of Detection , Seawater , Spectrophotometry, Atomic
5.
Anal Chim Acta ; 1191: 339282, 2022 Jan 25.
Article in English | MEDLINE | ID: mdl-35033257

ABSTRACT

Accurate detection of circulating microRNAs (miRNAs) plays a vital role in the diagnosis of various diseases. However, enzyme-free amplification detection remains challenging. Here, we report an enzyme-free fluorescence resonance energy transfer assay termed "3C-TASK" (cyclic click chemical-triggered hairpin stacking kit) for the detection of circulating miRNA. In this strategy, the miRNA could initiate copper-free click chemical ligation reactions and the ligated products then trigger another hairpin stacking circuit. The first signal amplification was achieved through the recycling of the target miRNA in the click chemical ligation circuit, and the second signal amplification was realized through the recycling of ligated probes in a hairpin stacking circuit driven by thermodynamics. The two-step chain reaction event triggered by miRNAs was quantified by the fluorescence signal value so that accurate detection of target miRNA could be achieved. The 3C-TASK was easily controlled because no enzyme was involved in the entire procedure. Although simple, this strategy showed sensitivity with a detection limit of 8.63 pM and specificity for distinguishing miRNA sequences with single-base variations. In addition, the applicability of this method in complex biological samples was verified by detecting target miRNA in diluted plasma samples. Hence, our method achieved sensitive and specific detection of miRNA and may offer a new perspective for the broader application of enzyme-free chemical reaction and DNA circuits in biosensing.


Subject(s)
Biosensing Techniques , Circulating MicroRNA , MicroRNAs , DNA , Limit of Detection , MicroRNAs/genetics , Nucleic Acid Amplification Techniques
6.
Anal Chim Acta ; 1191: 339279, 2022 Jan 25.
Article in English | MEDLINE | ID: mdl-35033266

ABSTRACT

Exosomes are promising biomarkers for cancer screening, but the development of a robust approach that can sensitively and accurately detect exosomes remains challenging. In the present study, an aptasensor based on the multifunctional signal probe 10-benzyl-2-amino-acridone (BAA) was developed for the colorimetric and photoelectrochemical detection and quantitation of exosomes. Exosomes are captured by cholesterol DNA anchor-modified magnetic beads (MBs) through hydrophobic interactions. This capture process can be monitored under a confocal fluorescence microscope using BAA as the fluorescent signal probe. The aptamer modified copper oxide nanoparticles (CuO NPs) then bind to mucin 1 (MUC1) on the surface of the exosomes to form a sandwich structure (MBs-Exo-CuO NPs). Finally, the MBs-Exo-CuO NPs are dissolved in nitric acid to generate Cu2+, which inhibits the visible-light-induced oxidase mimic activity and photoelectrochemical activity of BAA simultaneously. The changes in absorbance and photocurrent intensities are directly proportional to the concentration of exosomes. In this dual-modal aptasensor, the colorimetric assay can achieve rapid screening and identification, which is especially useful for point-of-care testing. The UV-vis absorbance and photocurrent assays then provide quantitative information, with a limit of detection of 1.09 × 103 particles µL-1 and 1.38 × 103 particles µL-1, respectively. The proposed aptasensor thus performs dual-modal detection and quantitation of exosomes. This aptasensor provides a much-needed toolset for exploring the biological roles of exosomes in specific diseases, particularly in the clinical setting.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Exosomes , Acridones , Colorimetry , Limit of Detection
7.
Anal Chim Acta ; 1191: 339312, 2022 Jan 25.
Article in English | MEDLINE | ID: mdl-35033271

ABSTRACT

A compact evanescent wave detection platform (EWDP) is developed for the detection of fluorescence gold nanoclusters. The EWDP employs a simple optical system and a Si-based photodetector SOP-1000 assembly to improve the optical efficiency and detection sensitivity. A microfluidic sample cell is also used to decrease the amount of analyte to 200 µL (The volume of sample cell is really about 30 µL). On this basis, we design a strategy for detecting dopamine (DA) based on the photoinduced electron transfer (PET) quenching mechanism. By introduction of tyrosinase (TYR) during the detection, the testing time is shortened to 1 min. The fluorescence emission signal decreased dramatically and the quenching ratio (F0-F)/F0 is linearly related to the concentration of DA in the range of 0.03-60 µM with a detection limit of 0.03 µM. Additionally, this detection platform has potential applications for DA fast detection in the microsamples.


Subject(s)
Dopamine , Gold , Electron Transport , Limit of Detection , Monophenol Monooxygenase/metabolism , Spectrometry, Fluorescence
8.
Anal Chim Acta ; 1191: 339291, 2022 Jan 25.
Article in English | MEDLINE | ID: mdl-35033276

ABSTRACT

The pregnancy-associated glycoproteins (PAGs) have been widely used as biomarkers for the early diagnosis of pregnancy in cattle and sheep. This study aimed to obtain the single-stranded DNA aptamers that specifically bound to ovine pregnancy-associated glycoprotein 7 (ovPAG7) with high affinity (Kd = 9.8-32.4 nM) using real serum sample-assisted FluMag-systematic evolution of ligands by exponential enrichment (SELEX). Subsequently, the selected aptamers were applied to fabricate an ultrasensitive colorimetric aptasensor for ovPAG7 detection based on functionalized magnetic microparticles and hybridization chain reaction. Under the optimized conditions, the colorimetric aptasensor exhibited a broad linear range (0.2-500 ng mL-1), low detection limit (0.081 ng mL-1), good recovery rate (94.5-109.1%), and high repeatability (relative standard deviation of 4.02-8.16%) in ovPAG7-spiked serum. Furthermore, this aptasensor was applied to measure the ovPAG7 in serum samples of ewes for pregnancy diagnosis. Blood samples were collected from Chinese Merino ewes on days 22, 28 after artificial insemination (AI) for ovPAG7 detection, respectively. Transrectal ultrasonography diagnosis of pregnancy 45 days after AI was the reference (gold) standard for all PAG tests. Diagnostic sensitivity, specificity, and accuracy of the proposed aptasensor were 95.8, 87.5, and 92.5% at day 22 and 95.8, 90.6, and 93.7% at day 28, respectively. The degree of agreement (Kappa) between developed aptasensor and ultrasonography diagnosis 22 and 28 day after AI were higher than 0.8. These results illustrated that the aptasensor was proved to be a sensitive, reliable and cost-effective way of measuring PAG and might be a useful means of pregnancy detection in ewes.


Subject(s)
Aptamers, Nucleotide , SELEX Aptamer Technique , Animals , Cattle , Colorimetry , Female , Glycoproteins , Limit of Detection , Magnetic Phenomena , Pregnancy , Sheep
9.
Food Chem ; 374: 131778, 2022 Apr 16.
Article in English | MEDLINE | ID: mdl-35021580

ABSTRACT

Core-shell structured magnetic covalent-organic frameworks (Fe3O4@TaTp) were facilely synthesized based on one-step functionalization at room temperature and applied for magnetic solid-phase extraction of okadaic acid from seawater and shellfish prior to LC-MS/MS detection. Parameters, including adsorbent amount, extraction time, desorption solution, and desorption time which could affect the extraction efficiency, were respectively investigated. The developed methods demonstrated good linearity (R2 > 0.99), acceptable accuracy and good precision (<15%), and low limit of detection (0.5 pg·mL-1 for seawater and 0.04 µg·kg-1 for shellfish). The amount of the material used (1 mg for seawater and 5 mg for shellfish) and the time required (4 min for seawater and 15 min for shellfish) for extracting analyte from 5 mL of seawater and 2 g of shellfish are both greatly shortened compared with the previous reports. In addition, we successfully applied this method to real sample analysis.


Subject(s)
Metal-Organic Frameworks , Adsorption , Chromatography, Liquid , Limit of Detection , Magnetic Phenomena , Okadaic Acid/analysis , Seawater , Shellfish/analysis , Solid Phase Extraction , Tandem Mass Spectrometry
10.
Sensors (Basel) ; 22(1)2021 Dec 23.
Article in English | MEDLINE | ID: mdl-35009607

ABSTRACT

The concentration of 5-hydroxymethyl-2-furfural (HMF) is an important quality-related index in milk and milk products. Fast, cost-effective and environmentally friendly determination of HMF is of great significance in milk products control. In this study, a three-dimensional (3D) graphene-like surface (3DGrls) was successfully prepared within 5 min by an electrochemical amperometric pretreatment on a pencil graphite electrode (PGE). The fast-obtained 3D graphene-like surface increased the electrode surface area and enhanced the electron transfer capability without the addition of any harmful chemicals. The morphology and chemical composition of the obtained electrode were characterized by scanning electron microscope (SEM), X-ray photoelectron spectroscopy (XPS), Raman spectroscopy and electrochemical impedance spectroscopy (EIS). The results found that the electrochemical response to HMF at the prepared 3DGrls/PGE was 34 times higher than that at PGE. The modified electrode showed a good linear response to HMF in a concentration range of 0.35~116 µM with a low limit of detection (LOD) of 0.099 µM. The integrated electrode also exhibited excellent stability and wonderful antifouling property. Furthermore, the 3DGrls/PGE was successfully applied for the determination of HMF in three processed cheese samples with satisfactory results.


Subject(s)
Cheese , Graphite , Electrochemical Techniques , Electrodes , Furaldehyde/analogs & derivatives , Limit of Detection
11.
Sensors (Basel) ; 22(1)2021 Dec 24.
Article in English | MEDLINE | ID: mdl-35009659

ABSTRACT

In this work, the development of an electrochemical sensor for melatonin determination is presented. The sensor was based on Sonogel-Carbon electrode material (SNGCE) and Au nanoparticles (AuNPs). The low-cost and environmentally friendly SNGCE material was prepared by the ultrasound-assisted sonogel method. AuNPs were prepared by a chemical route and narrow size distribution was obtained. The electrochemical characterization of the SNGCE/AuNP sensor was carried out by cyclic voltammetry in the presence of a redox probe. The analytical performance of the SNGCE/AuNP sensor in terms of linear response range, repeatability, selectivity, and limit of detection was investigated. The optimized SNGCE/AuNP sensor displayed a low detection limit of 8.4 nM melatonin in synthetic samples assessed by means of the amperometry technique. The potential use of the proposed sensor in real sample analysis and the anti-matrix capability were assessed by a recovery study of melatonin detection in human peripheral blood serum with good accuracy.


Subject(s)
Melatonin , Metal Nanoparticles , Carbon , Electrochemical Techniques , Electrodes , Gold , Humans , Limit of Detection
12.
Sensors (Basel) ; 22(1)2021 Dec 30.
Article in English | MEDLINE | ID: mdl-35009811

ABSTRACT

Tetracycline (TC) is a widely known antibiotic used worldwide to treat animals. Its residues in animal-origin foods cause adverse health effects to consumers. Low-cost and real-time measuring systems of TC in food samples are, therefore, extremely needed. In this work, a three-electrode sensitive and label-free sensor was developed to detect TC residues from milk and meat extract samples, using CO2 laser-induced graphene (LIG) electrodes modified with gold nanoparticles (AuNPs) and a molecularly imprinted polymer (MIP) used as a synthetic biorecognition element. LIG was patterned on a polyimide (PI) substrate, reaching a minimum sheet resistance (Rsh) of 17.27 ± 1.04 Ω/sq. The o-phenylenediamine (oPD) monomer and TC template were electropolymerized on the surface of the LIG working electrode to form the MIP. Surface morphology and electrochemical techniques were used to characterize the formation of LIG and to confirm each modification step. The sensitivity of the sensor was evaluated by differential pulse voltammetry (DPV), leading to a limit of detection (LOD) of 0.32 nM, 0.85 nM, and 0.80 nM in buffer, milk, and meat extract samples, respectively, with a working range of 5 nM to 500 nM and a linear response range between 10 nM to 300 nM. The sensor showed good LOD (0.32 nM), reproducibility, and stability, and it can be used as an alternative system to detect TC from animal-origin food products.


Subject(s)
Graphite , Metal Nanoparticles , Molecular Imprinting , Animals , Anti-Bacterial Agents , Electrochemical Techniques , Electrodes , Gold , Lasers , Limit of Detection , Meat , Milk , Molecularly Imprinted Polymers , Polymers , Reproducibility of Results , Tetracycline
13.
Biosens Bioelectron ; 200: 113926, 2022 Mar 15.
Article in English | MEDLINE | ID: mdl-34990956

ABSTRACT

In this work, an unprecedented study exploring the role that slight changes into the Pd/Au proportion have in the electrocatalytic activity of bimetallic Pd-AuNPs toward the oxygen reduction reaction (ORR) is conducted. In particular, a careful control of the amount of Au atoms introduced in the cluster and the evaluation of the optimum Pd:Au ratio for getting the maximum catalytic activity is performed for the first time. First, PdNPs are synthesized by alcohol reduction in the presence of polyvinylpyrrolidone, and gold atoms are selectively introduced on vertex or corner positions of the cluster in different amounts following a galvanic substitution procedure. Average elemental analysis done relying on EDX spectroscopy allows to evaluate the Pd:Au ratio in the Pd-AuNPs obtained. Lineal sweep voltammetry and chronoamperometry are used for the evaluation of the Pd-AuNPs electrocatalytic activity toward ORR at a neutral pH compared to PdNPs and AuNPs alone. Our results indicate that, the synergy between both metals is strongly enhanced when the amount of gold is controlled and occupies the more reactive positions of the cluster, reaching a maximum activity for the NPs containing a 30% of gold, while an excess of this metal leads to a decrease in such activity, as a shelter of the PdNPs is achieved. Chronoamperometric analysis allows the quantification of the optimal Pd-AuNPs at over 6 × 109 NPs/mL levels. Such optimal Pd-AuNPs were used as tags, taking advantage of the bio-functionalities of gold present in the cluster, in a proof-of-concept electrochemical immunosensor for the detection of hyaluronidase wound infection biomarker, using magnetic beads as platforms. Hyaluronidase was detected at levels as low as 50 ng/mL (0.02 U/mL; 437 U/mg) with good reproducibility (RSD below 8%) and selectivity (evaluated against bovine serum albumin, immunoglobulin G and lysozyme). The low matrix effects inherent to the use of magnetic bead platforms allowed us to discriminate between wound exudates with both sterile and infected ulcers without sample pre-treatment. This novel electrocatalytic immunoassay has the advantage, over common methods for NP tags electrochemical detection, of the signal generation in the same neutral medium where the immunoassay takes place (10 mM PBS pH 7.4), avoiding the use of additional and hazardous reagents, bringing it closer to their use as point-of-care devices. Overall, our findings may be of great interest not only for biosensing, but also for applications such as energy converting on fuel cells, in which the ORR has a pivotal role.


Subject(s)
Biosensing Techniques , Metal Nanoparticles , Wound Infection , Biomarkers , Electrochemical Techniques , Gold , Humans , Immunoassay , Limit of Detection , Palladium , Reproducibility of Results
14.
Biosens Bioelectron ; 200: 113922, 2022 Mar 15.
Article in English | MEDLINE | ID: mdl-34990959

ABSTRACT

Fast, affordable, portable, and sensitive technology to detect COVID-19 is critical to address the current outbreak. Here, we present a CRISPR/Cas12a-derived electrochemical aptasensor for cost-effective, fast, and ultrasensitive COVID-19 nucleocapsid protein (Np) detection. First, an electrochemical sensing interface was fabricated by immobilizing methylene blue labeled poly adenines DNA sequence (polyA-MB electrochemical reporter) on a gold electrode surface. Second, an arched probe was prepared via hybridization of Np aptamer and an activator strand. In the presence of COVID-19 Np, the activator strand could be released from the arched probe due to the specific interaction between the target and the aptamer, which then activated the trans-cleavage activity of the CRISPR/Cas12a system. Subsequently, the polyA-MB reporters were cleaved from the electrode surface, decreasing the current of differential pulse voltammetry (DPV) at a potential of -0.27 V(vs. Ag/AgCl). The CRISPR/Cas12a-derived electrochemical aptasensor shows a highly efficient performance for COVID-19 Np detection in 50 pg mL-1 to 100 ng mL-1 with a limit of detection (LOD) low to 16.5 pg mL-1. Notably, the whole process of one test can be completed within 30 min. Simultaneously, the aptasensor displays a high selectivity to other proteins. The further measurements demonstrate that the aptasensor is robust in a natural system for point-of-care testing, such as in tap water, milk, or serum. The aptasensor is universal and expandable and holds great potential in the COVID-19 early diagnosis, environmental surveillance, food security, and other aspects.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , COVID-19 , CRISPR-Cas Systems , Electrochemical Techniques , Electrodes , Gold , Humans , Limit of Detection , Nucleocapsid Proteins , SARS-CoV-2
15.
Environ Monit Assess ; 194(2): 58, 2022 Jan 06.
Article in English | MEDLINE | ID: mdl-34989878

ABSTRACT

This study utilized switchable solvent liquid-phase microextraction (SS-LPME) to enrich eleven nervous system active pharmaceutical ingredients (APIs) from aqueous samples for their determination at trace levels by gas chromatography mass spectrometry. The analytes selected for the study included APIs utilized in antidepressant, antipsychotic, antiepileptic, and anti-dementia drugs. Parameters of the microextraction method including switchable solvent volume, concentration and volume of the trigger agent (sodium hydroxide), and sample agitation period were optimized univariately to boost extraction efficiency. Under the optimum conditions, the detection limits calculated for the analytes were in the range of 0.20-8.0 ng/mL, and repeatability for six replicate measurements as indicated by percent relative standard deviation values were below 10%. Matrix matching calibration strategy was used to enhance quantification accuracy for the analytes. The percent recovery results calculated for the eleven analytes ranged between 86 and 117%.


Subject(s)
Liquid Phase Microextraction , Pharmaceutical Preparations , Water Pollutants, Chemical , Calibration , Environmental Monitoring , Gas Chromatography-Mass Spectrometry , Limit of Detection , Nervous System/chemistry , Solvents , Water Pollutants, Chemical/analysis
16.
J Biomed Nanotechnol ; 17(12): 2444-2454, 2021 Dec 01.
Article in English | MEDLINE | ID: mdl-34974867

ABSTRACT

An electrochemical aptasensor for quantitatively detecting glypican-3 (GPC3) was constructed by combining hemin-reduced graphene oxide-platinum (H-rGO-Pt) nanoparticles (NPs) with reduced graphene oxide-gold (rGO-Au) nanoparticles (NPs). Herein, the rGO-Au NPs deposited onto screen-printed electrodes resulted in signal amplification due to their large surface areas. Meanwhile, highly conductive H-rGO-Pt NPs acted as a sensing medium that improved electrical conductivity and as an indicator for monitoring peak current for determination. A GPC3 aptamer (GPC3apt) with a low equilibrium dissociation constant was used as a bio-recognition molecule. GPC3apt specifically captured GPC3 proteins and formed aptamer-GPC3 complexes, which impeded electron transfer and thus hampered the redox signal of hemin in H-rGO-Pt NPs. This developed electrochemical aptasensor showed a linear response to GPC3 (from 0.001 µg/mL to 10 µg/mL) and had a detection limit of 0.001 µg/mL. This work provides a low-cost and highly sensitive detection with and good recovery for GPC3 and holds great promise for the clinical diagnosis of hepatocellular carcinoma.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Graphite , Metal Nanoparticles , Electric Conductivity , Electrochemical Techniques , Glypicans , Gold , Hemin , Limit of Detection , Platinum
17.
J Biomed Nanotechnol ; 17(12): 2495-2504, 2021 Dec 01.
Article in English | MEDLINE | ID: mdl-34974872

ABSTRACT

An aptasensor was developed on an interdigitated microelectrode (IDME) by current-volt sensing for the diagnosis of ulcerative colitis by detecting the biomarker lipocalin-2. Higher immobilization of the anti-lipocalin-2 aptamer as a probe was achieved by using sodium dodecyl benzenesulfonate-aided zeolite particles. FESEM and FETEM observations revealed that the size of the zeolite particles was <200 nm, and they displayed a uniform distribution and spherical shape. XPS analysis attested the occurrence of Si, Al, and O groups on the zeolite particles. Zeolite particles were immobilized on IDME by a (3-aminopropyl)-trimethoxysilane amine linker, and then, the aptamer as the probe was tethered on the zeolite particles through a biotin-streptavidin strategy assisted by a bifunctional aldehyde linker. Due to the high occupancy of the aptamer and the efficient electric transfer from zeolite particles, higher changes in current can be observed upon interaction of the aptamer with lipocalin-2. The lower detection of lipocalin-2 was noted as 10 pg/mL, with a linear range from 10 pg/mL to 1 µg/mL and a linear regression equation of y=8E-07x+8E-08; R² = 0.991. Control experiments with complementary aptamer and matrix metalloproteinase-9 indicate the specific detection of lipocalin-2. Furthermore, spiking lipocalin-2 in human serum does not interfere with the identification.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Colitis, Ulcerative , Nanoparticles , Zeolites , Colitis, Ulcerative/diagnosis , Electrodes , Humans , Limit of Detection , Lipocalin-2 , Microelectrodes
18.
J Chromatogr A ; 1663: 462768, 2022 Jan 25.
Article in English | MEDLINE | ID: mdl-34974368

ABSTRACT

New psychoactive substances (NPS) continue to emerge in the drug market every year, becoming a global threat to public health and safety. These compounds are mostly synthetic cannabinoids and designer cathinones. However, synthetic opioids have appeared on the recreational drug markets in recent years, particularly fentanyl and its derivatives ("fentanyls"). Fentanyl and its analogs are related to harmful intoxications and an increase in opioid-related mortality in many countries, such as in the United States and Europe in the last years. Taking the drug related global crisis into consideration, this work developed and validated an effective and sensitive method based on fabric phase sorptive extraction (FPSE) followed by gas chromatography-mass spectrometry (GC-MS) for the simultaneous determination of 11 fentanyl analogs in oral fluid samples. The extraction was carried out using a sol-gel Carbowax 20 M sorbent immobilized on 100% cellulose fabric substrate and using ethyl acetate as the desorption solvent. The limits of detection (LODs) and quantification (LOQs) ranged from 1 to 15 ng mL-1 and 5 to 50 ng mL-1, respectively. Intra-day and inter-day precision were found within 8.2% and 8.6%, respectively, while accuracy ranged from -5.5 to 9.1%, in accordance with the established criteria. The absolute recovery values were in the range of 94.5%-109.1%. The validated method demonstrated its great potential to detect and quantify fentanyl analogs in possible forensic work and off-site analysis in road traffic cases.


Subject(s)
Analgesics, Opioid , Illicit Drugs , Gas Chromatography-Mass Spectrometry , Limit of Detection , Textiles
19.
J Chromatogr A ; 1663: 462743, 2022 Jan 25.
Article in English | MEDLINE | ID: mdl-34974369

ABSTRACT

Synthetic cannabinoids (SCs) are new psychoactive substances that function as endocannabinoid CB1 and CB2 receptor agonists. Abuse of SCs can lead to symptoms such as confusion, dizziness, and even death. At present, Synthetic cannabinoids constitute one of the largest groups of new psychoactive substances and become popular recreational drugs of abuse for their psychoactive properties. The continuous transformation of SCs also leads to an endless emergence of new types. An efficient, high-throughput screening method is therefore very important for their identification. This paper describes a liquid chromatography-high resolution mass spectrometry (LC-HRMS) method for simultaneously screening 179 SCs and 80 SC metabolites in blood and urine. Simple acetonitrile was used to precipitate the blood and urine proteins, and the supernatants obtained after centrifugation were analyzed. The LC-HRMS run time was 20 min. The mass spectrometer used an ESI source with a scanning range of m/z 100-1000. LC-HRMS provided accurate mass, retention time, and fragment ions for qualitative analysis. The method validation results showed that the limits of detection (LODs) for over 80% compounds were 5 ng/mL in blood and urine samples. At low concentrations (50 ng/mL), 229 compounds (95.8%) in the blood showed recoveries of more than 50%, and 232 compounds (97.1%) had matrix effects greater than 80%. In the urine, 219 compounds (91.6%) had recoveries above 50%, and the matrix effects of 234 compounds (97.9%) were greater than 80%. This method was successfully applied to actual forensic cases.


Subject(s)
Cannabinoids , Illicit Drugs , Chromatography, Liquid , Limit of Detection , Mass Spectrometry , Substance Abuse Detection
20.
Food Chem ; 375: 131888, 2022 May 01.
Article in English | MEDLINE | ID: mdl-34974348

ABSTRACT

In this study, an ultrasensitive monoclonal antibody (mAb) was produced and used to develop a gold nanoparticle-based lateral flow immunochromatographic (ICA) strip for screening of clomazone (CLO) in potato and pumpkin samples. With assayed by indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) method, the mAb belonging of IgG2 subclass showed a half-maximal inhibitory concentration (IC50) of 3.47 ng/mL and a linear range of detection of 0.43-28.09 ng/mL. A cross-reactivity test revealed that the mAb had good specificity for CLO. The strip assay had a visual limit of detection (LOD) of 5 µg/kg and a cut-off value of 50 µg/kg for CLO pumpkin samples (potato samples was 100 µg/kg) when evaluated with the naked eye. The results were consistent with ic-ELISA and high performance liquid chromatography tandem mass spectrometry (HPLC-MS). Thus, this ICA strip assay represents a potentially tool for on-site and rapid initial detection of CLO in potato and pumpkin samples.


Subject(s)
Cucurbita , Metal Nanoparticles , Solanum tuberosum , Enzyme-Linked Immunosorbent Assay , Gold , Gold Colloid , Immunoassay , Isoxazoles , Limit of Detection , Oxazolidinones
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