Dual-potential electrochemiluminescence cytosensor based on a metal-organic framework and ABEI-PEI-Au@AgNPs for the simultaneous determination of phosphatidylserine and epidermal growth factor receptors on an apoptotic cell surface.

A new electrochemiluminescence (ECL) cytosensor is proposed for the simultaneous determination of phosphatidylserine (PS) and epidermal growth factor receptor (EGFR) based on the ECL signals of metal-organic framework-5 (MOF-5) loaded CdS quantum dots and N-(aminobutyl)-N-(ethylisoluminol)-polyethylenimine capped Au and Ag nanoparticles. Apoptosis promotes the exposure of PS and reduces the expression of EGFR in cell membranes. Two spatially resolved areas on dual-disk glassy carbon electrodes were designed to eliminate the interference from different ECL probes. Using HepG2 cells treated with resveratrol to induce apoptosis, the cytosensor exhibited high sensitivity, simplicity, and high reproducibility, demonstrating its potential in drug screening and rapid apoptotic cell detection. The strategy reported provides a promising platform for the highly sensitive cytosensing and convenient screening of clinically relevant anticancer drugs.

Electrochemiluminescence resonance energy transfer of MnCO3 for ultrasensitive amyloid-ß protein detection.

A composite material MnCO3/poly(diallyl dimethyl ammonium chloride) (PDDA)/Ag with excellent electrochemiluminescence (ECL) performance and high biocompatibility was prepared by adding MnCO3 and PDDA to silver nanoparticles (AgNPs). MnCO3/PDDA/Ag and Au@SiO2NPs were used as ECL donors and acceptors, respectively. Thus, an effective ECL-resonance energy transfer (RET) sensing platform was established. In a potassium persulfate (K2S2O8) medium, MnCO3 exhibited ECL emission with an ECL band appearing at 500-600 nm. In addition, Au@SiO2 nanoparticles showed a UV-visible absorption at 450-650 nm. The ECL emission spectra of MnCO3 overlapped with the absorption spectra of Au@SiO2NPs. The effective ECL quenching resulted in a good response to the concentration of Aß42 in serum samples. The linear range was 5 fg ⋅ mL-1 to 100 ng ⋅ mL-1, and the detection limit was 2 fg ⋅ mL-1. The recovery ranged from 97.7% to 104%. The high-efficiency ECL-RET immunosensor has potential application in detecting human serum Aß42 and other biomarkers, and can be used for the early screening of diseases.

Spatially-resolved dual-potential sandwich electrochemiluminescence immunosensor for the simultaneous determination of carbohydrate antigen 19-9 and carbohydrate antigen 24-2.

A new electrochemiluminescence (ECL) immunosensor based on spatially-resolved dual-potential technology was designed for the simultaneous determination of carbohydrate antigen 19-9 (CA 19-9) and carbohydrate antigen 24-2 (CA 242). Luminol-AgNPs@ZIF-67 was used as the anodic probe, and Pt nanoparticle-functionalized graphitic carbon nitride nanosheets (g-C3N4@PtNPs) were used as the cathodic probe. Two spatially-resolved areas on the dual-disk glassy carbon electrode (DDGCE) were modified with a AuNPs film by electrodeposition to improve the conductivity of the sensing interface. By recording the ECL responses at two different excitation potentials, the linear range for CA 19-9 was determined to be 0.0001-10 U/mL, with a limit of detection of 31 µU/mL. The linear range for CA 242 was 0.0005-10 U/mL, with a limit of detection of 0.16 mU/mL. Moreover, the ECL immunosensor possessed high selectivity and stability and successfully detected CA 19-9 and CA 242 in real samples. This immunosensor provides a new platform for clinical immunoassays.

A potential-resolved electrochemiluminescence resonance energy transfer strategy for the simultaneous detection of neuron-specific enolase and the cytokeratin 19 fragment.

An electrochemiluminescence resonance energy transfer (ECL-RET) immunosensor was developed based on the potential-resolved technology for the simultaneous detection of neuron-specific enolase (NSE) and the cytokeratin 19 fragment (CYFRA21-1). The absorption spectrum of gold nanorods (AuNRs) perfectly overlapped with the ECL spectra of SnS2@Pt and Ru(bpy)32+/Zn-MOF, so they exhibited an excellent ECL-RET effect with high efficiency. Zn-MOF possesses a large surface area, which allows for the loading of Ru(bpy)32+. This results in a signal probe of Ru(bpy)32+/Zn-MOF/Ab1 showing a strong ECL emission. Simultaneously, owing to the excellent electronic conductivity of PtNPs, they can increase the electron transfer rate between S2O82- and tin disulfide nanoflowers (SnS2NFs). Hence, the ECL signal of SnS2NFs can be enhanced. Under the optimal conditions, the linear range for NSE is 0.2 pg mL-1-20 ng mL-1 with a detection limit of 79 fg mL-1. The linear range for CYFRA21-1 is 1.25 pg mL-1-12.5 ng mL-1 with a detection limit of 0.43 pg mL-1. The proposed immunosensor can be used for the sensitive simultaneous detection of NSE and CYFRA21-1 in human serum and has promise for clinical diagnostics.

Electrochemiluminescence immunoassay of human chorionic gonadotropin using silver carbon quantum dots and functionalized polymer nanospheres.

A composite, reduced graphene oxide (rGO) doped with silver nanoparticles (Ag NPs), was prepared by using binary reductants of sodium citrate and hydrazine hydrate. Carbon quantum dots (CQDs) synthesized by papaya peel combined with silver ions to form a CQDs-loaded silver nanoparticle (AgCQDs) nanocomposite. Polymer nanospheres (PNS) were generated via the infinite coordination polymer of ferrocene dicarboxylic acid and employed as carriers to load AgCQDs. The prepared AgCQDs@PNS-PEI has good biocompatibility and electrical conductivity and can be used as a matrix for the immobilization of a secondary antibody (Ab2). A sandwich-type electrochemiluminescence (ECL) immunosensor using AgCQDs@PNS-PEI nanocomposite as probe has been developed for the detection of human chorionic gonadotropin (HCG). The proposed immunosensor exhibits a linear range from 0.00100 to 500 mIU mL-1 and the detection limit is 0.33 µIU mL-1 (S/N = 3) under optimal conditions. The sensor exhibits excellent selectivity, good reproducibility, and high stability. These features demonstrate that the proposed method has promising potential for clinical protein detection and displays a new strategy to fabricate an immunosensor. Graphical abstract.

[Selenium speciation in watermelon by g-C3N4 enrichment combined with capillary electrophoresis-inductively coupled plasma-mass spectrometry].

Selenium is one of the essential trace elements in the human body, and it plays a critical role in human health. In this work, 2.0 g melamine was placed in an alumina crucible, which was heated in a box-type resistance furnace for 2 h at 600 ℃, at the heating rate of 3 ℃/min, and then cooled to room temperature. After cooling, yellow graphite phase carbon nitride (g-C3N4) nanosheets were obtained. Subsequently, 500 mg of the nanosheets was dispersed in 50 mL water with ultrasonication for 10 h in order to remove the residual un-exfoliated g-C3N4 nanoparticles and large-sized nanosheets. The obtained suspension was centrifuged at about 10000 r/min, followed by drying at 60 ℃ to produce g-C3N4. The prepared g-C3N4 was characterized by Fourier transform infrared spectroscopy (FT-IR), X-ray powder diffraction (XRD), and field emission-environmental scanning electron microscopy (SEM) analyses. Given that the selenium content in actual samples is very low, high sensitivity, and accuracy are imperative for selenium detection. The combination of capillary electrophoresis (CE) with inductively coupled plasma-mass spectrometry (ICP-MS) can greatly improve the sensitivity, accuracy, and speed of the analysis. A novel method based on CE-ICP-MS was established for the determination of selenourea (SeUr), L-selenocystine (SeCys2), DL-selenomethionine (SeMet), selenite (Se(Ⅳ)), selenate (Se(Ⅵ)), and selenoethionine (SeEt) in watermelon. The selenium species in watermelon were extracted by ultrasonication with pepsin as an extractant and g-C3N4 enrichment. The enrichment factor of g-C3N4 ranged from 12 to 29. Six selenium species were completely separated within 11 min in a 100-cm-long capillary with 100 µm internal diameter, at an applied voltage of 22 kV, using a buffer solution of 8 mmol/L NaH2PO4-12 mmol/L H3BO3-0.2 mmol/L cetyl trimethyl ammonium bromide (CTAB; pH 9.2). The interference in the selenium detection was eliminated using a dynamic reaction cell with CH4. The linear correlation coefficients of all the selenium species were greater than 0.9995. Under the optimal conditions, the limits of detection (3 σ, σ for standard deviation, as Se) for SeUr, SeCys2, SeMet, Se(Ⅳ), Se(Ⅵ), and SeEt were 6.2, 30, 11, 8.2, 48, and 5.5 ng/L, respectively. The linear range (as Se) for SeUr, SeCys2, SeMet, Se(Ⅳ), Se(Ⅵ), and SeEt were 0.017-20 µg/L, 0.091-50 µg/L, 0.032-40 µg/L, 0.023-60 µg/L, 0.015-75 µg/L, and 0.015-30 µg/L, respectively. The recoveries ranged from 96.0% to 106%, and the relative standard deviations (RSDs; n=5) were less than 3%. The developed method is simple, rapid, and sensitive, and it is also suitable for the detection of selenium species in other food and environmental samples.

A highly efficient introduction system for single cell- ICP-MS and its application to detection of copper in single human red blood cells.

A method of simultaneous cell counting and determination of metals in single cells using time-resolved inductively coupled plasma-mass spectrometry (ICP-MS) was reported. A facile, low cost and highly efficient single-cell introduction system of time-resolved ICP-MS consists of a flow cell, a visual contrast calibration device, a customized nebulizer and a fabricated spray chamber. The flow cell includes a cell sample tube, a sheath liquid tube and a flow chamber. The visual contrast calibration device was composed of a microscope with a 16 × microscope objective (160 × total magnification). The flow chamber was used to combine a flow of red blood cell suspension (0.800 µL/min) and a flow of PBS (4.40 µL/min) into the nebulizer. The intact cells were directly introduced with the single-cell introduction system into the plasma via nebulizing, and then ion plumes corresponding to single cells were individually detected with mass spectrometer. The frequency of the spikes directly reflects the number of cells, and the intensity of spikes is proportional to the concentration of copper within one cell. The single-cell introduction system can be transported into the ICP-MS via a customized transport system with 100% efficiency. A high cell introduction efficiency into the plasma supports for a reduction of cell consumption. The Cu signal frequency was about 120 cell events per minute. This single-cell introduction system simplifies the introduction of individual and intact cells. The copper content in single red blood cell was 0.20-0.40 fg.

Electrochemiluminescent immunoassay for neuron specific enolase by using amino-modified reduced graphene oxide loaded with N-doped carbon quantum dots.

An ultrasensitive electrochemiluminescence based sandwich immunoassay is presented for determination of neuron specific enolase. The method uses silver-cysteine nanowires as the capture probe and a composite made of amino-modified reduced graphene oxide and nitrogen-doped carbon quantum dots as the signal probe. It was synthesized by covalent coupling of amino-modified reduced graphene oxide to the carboxy groups of nitrogen-doped carbon quantum dots. The nanowires possess a large specific surface and abundant functional groups which facilitate immobilizing the primary antibody (Ab1). The amino-modified reduced graphene oxide is employed as a carrier for loading a large number of the quantum dots and secondary antibody (Ab2). This increases the electrochemiluminescence intensity of quantum dots. Response to neuron specific enolase is linear in the 0.55 fg·mL-1 to 5.5 ng·mL-1 concentration range. It has a detection limit of 0.18 fg·mL-1 (at S/N = 3). The relative standard deviation (for n = 6) is less than 2.9%. The assay is highly sensitive, reproducible, selective and stable. Graphical abstractA novel electrochemiluminescence immunosensor is described that uses amino-modified reduced graphene oxide (amino-rGO), nitrogen-doped carbon quantum dots (N-CQDs) and silver-cysteine nanowires (SCNWs). It was applied to the determination of neuron specific enolase (NSE). Bovine serum albumin: BSA;1-ethyl-3-(3-dimethylaminopropyl)carbodiimide: (EDC;, N-hydroxysuccinimide: NHS.

Facile Preparation of Boron and Nitrogen Codoped Green Emission Carbon Quantum Dots for Detection of Permanganate and Captopril.

A hydrothermal strategy for preparing boron and nitrogen codoped carbon quantum dots was studied using the precursors of p-amino salicylic acid, boric acid and ethylene glycol dimethacrylate. The boron and nitrogen codoped carbon quantum dots have high fluorescence intensity, good monodispersity, high stability, superior water solubility, and a fluorescence quantum yield of 19.6%. Their average size is 5 nm. Their maximum excitation and emission wavelengths are 380 and 520 nm, respectively. Permanganate (MnO4-) quenched boron and nitrogen codoped carbon quantum dots fluorescence through inner filter effect and static quenching effects. The linear relation between quenching efficiency and MnO4- concentration ranged from 0.05 to 60 µmol/L with a detection limit of 13 nmol/L. In the presence of captopril, MnO4- was reduced to Mn2+ and the fluorescence of boron and nitrogen codoped carbon quantum dots was recovered. The linear range between recovery and captopril concentration was from 0.1 to 60 µmol/L. The limit of detection was 0.03 µmol/L. The developed method can be employed as a sensitive fluorescence sensing platform for MnO4-. It has been successfully used for captopril detection in mouse plasma.

A Novel Carbon Quantum Dots Signal Amplification Strategy Coupled with Sandwich Electrochemiluminescence Immunosensor for the Detection of CA15-3 in Human Serum.

A sensitive sandwich electrochemiluminescence immunosensor was established by employing graphene oxide-PEI-carbon quantum dots (CQDs)-Au nanohybrid as probe to measure carbohydrate antigen 15-3 (CA15-3), a breast cancer biomarker. In this work, nanocomposites of Ag nanoparticles and polydopamine (AgNPs-PDA) were synthesized by redox reaction between dopamine and Ag+. The nanocomposite with high surface area can provide an efficient substrate for immobilizing initial antibody (Ab1). Carbon quantum dots (CQDs) are fixed on polyethylenimine-functionalized graphene oxide (PEI-GO) by amide bonds. Au nanoparticles are modified on CQDs-decorated PEI-GO substrates. The secondary antibody (Ab2) was immobilized by AuNPs/CQDs-PEI-GO composite. CQDs can be assembled onto the surface of an electrode by incorporation of CA15-3 with Ab1 and Ab2. Under the synergistic action of AgNPs, polydopamine, AuNPs, and PEI-GO, the ECL signal of CQDs is greatly amplified as an excellent conductive material to facilitate electron transfer rate and further increase electrochemical detection capability. Under optimal conditions, the fabricated immunosensor showed a linear concentration range from 0.005 to 500 U mL-1, with a detection limit of 0.0017 U mL-1 (signal-to-noise ratio of 3) for CA15-3. The designed ECL immunosensor displayed receivable accuracy, excellent stability, and high specificity. The results of the detection of human serum samples are satisfactory, revealing that the method offers a potential application for the clinical diagnosis of tumor markers.

Based on reduced graphene oxide-copper sulfide-carbon nitride nanosheets composite electrochemiluminescence sensor for determination of gatifloxacin in mouse plasma.

A new method for determination of gatifloxacin hydrochloride (GAT) by reduced graphene oxide-copper sulfide (rGO-CuS) composite coupled with graphite-like carbon nitride nanosheets (g-C3N4 NSs) modified glassy carbon electrode was developed. In this work, rGO-CuS composite was synthesized by one-pot hydrothermal method and used for enhancing sensitivity of GAT. g-C3N4 NSs were synthesized as radiant agent. The sensor characteristics of electrochemistry and electrochemiluminescence (ECL) were investigated. The ECL intensity has enhanced four-fold after modifying with rGO-CuS composite. The results can be ascribed to the presence of rGO-CuS composite on the electrode surface that facilitates the electron transfer rate between the electroactive center of g-C3N4 NSs and the electrode. Under the optimum experimental conditions (photomultiplier tuber for 800 V, scan rate for 0.1 V/s, 0.1 mol/L PBS (pH 7.5) and 1.0 × 10-2 mol/L K2S2O8), the linear range for GAT was from 1.0 × 10-4 to 1.0 × 10-8 mol/L (R2 = 0.9991) with detection limit of 3.5 × 10-9 mol/L (S/N = 3). RSD for ECL intensity was 4.8% (n = 10). The recoveries of GAT in mouse plasma samples were from 98.36 to 104.7%. The sensor showed the advantages of low cost, high sensitivity and wide application in drug analysis.

A novel ECL sensor based on a boronate affinity molecular imprinting technique and functionalized SiO2@CQDs/AuNPs/MPBA nanocomposites for sensitive determination of alpha-fetoprotein.

In this work, a boronate-affinity sandwich electrochemiluminescence (ECL) sensor was constructed to detect alpha-fetoprotein (AFP) based on a multiple signal amplification strategy. Gold nanoparticles (AuNPs) were utilized and modified on the surface with chitosan in order to facilitate electron transfer. The composite of the molecularly imprinted polymer (MIP) enhanced the selectivity of alpha-fetoprotein detection. 4-mercaptophenylboronic acid (MPBA) was used as the tracing tag for capture of alpha-fetoprotein. SiO2 nanoparticles carried carbon quantum dots (CQDs) labeled with gold nanoparticles and produced an ECL signal. Under the optimum experimental conditions, the linear range for alpha-fetoprotein was between 0.001 and 1000 ng/mL with a correlation coefficient of 0.9952, and the detection limit was 0.0004 ng/mL (S/N = 3). This proposed ECL sensor displayed several advantages, including outstanding selectivity, fine reproducibility, high sensitivity, low detection limit and wide linear range. Furthermore, the newly constructed boronate-affinity sandwich ECL sensor was successfully applied to the determination of alpha-fetoprotein in serum samples, indicating great potential for application in clinical diagnostics.

Ginkgo leaf-based synthesis of nitrogen-doped carbon quantum dots for highly sensitive detection of salazosulfapyridine in mouse plasma.

A simple, economical hydrothermal strategy for synthesizing nitrogen-doped carbon quantum dots (N-CQDs) was developed using Ginko leaves as a carbon source. These N-CQDs have strong blue fluorescence, excitation-relevant emissions, high monodispersity, good stability, good water solubility, and a 22.8% fluorescence quantum yield. They average 3 nm in size, and have maximum excitation and emission wavelengths of 350 and 436 nm, respectively. They are used as an effective fluorescent sensing platform for the label-free sensitive detection of salazosulfapyridine (SASP) due to the strong quenching effect of SASP. When SASP concentration is 0.1-80 µmol/L, there is a good linear relationship with a detection limit of 40 nmol/L. This method was successfully applied to detect SASP in mouse plasma. The results show that the SASP recovery range was 96%-101%. RSDs ranged from 2.6% to 3.1%.

Based on ZnSe quantum dots labeling and single particle mode ICP-MS coupled with sandwich magnetic immunoassay for the detection of carcinoembryonic antigen in human serum.

A highly sensitive sandwich-type magnetic immunoassay based on inductively coupled plasma mass spectrometry detection in single particle mode, with ZnSe Quantum dots (QDs) serving as model tags, was proposed. The transient signals induced by the flash of ions (64Zn+) in the plasma torch from the ionization of nanoparticles tagged on antibody were recorded in a single particle mode. The frequency of transient signals is directly related to the concentration of nanoparticle tags, and the concentration of nanoparticle tagged antibodies can be quantified by the frequency of transient signals. Amino-modified magnetic nanoparticles (AMNPs) were synthesized and conjugated with primary carcinoembryonic antigen (CEA) antibody to extract the target biomarker. ZnSe QDs were synthesized as a probe to determine CEA by ICP-MS. A detection limit of 0.006 ng mL-1 was obtained for CEA after immunoreactions, and a wide linear range of 0.02-100 ng mL-1 with the relative standard deviation (RSD) was 4.4%. The method was successfully applied to human serum samples.