Dual-Response Ratiometric Electrochemical Microsensor for Effective Simultaneous Monitoring of Hypochlorous Acid and Ascorbic Acid in Human Body Fluids.

Redox homeostasis between hypochlorous acid (HClO/ClO-) and ascorbic acid (AA) significantly impacts many physiological and pathological processes. Herein, we report a new electrochemical sensor for the simultaneous determination of HClO and AA in body fluids. We first coated a carbon fiber microelectrode (CFME) with a three-dimensional nanocomposite consisting of graphene oxide (GO) and carbon nanotubes (CNTs) to fabricate the CFME/GO-CNT electrode. After the electrochemical reduction of GO (ERGO), we integrated a latent 1-(3,7-bis(dimethylamino)-10H-phenothiazin-10-yl)-2-methylpropan-1-one (MBS) electrochemical molecular recognition probe to monitor HClO and employed anthraquinone (AQ) as an internal reference. The compact CFME/ERGO-CNT/AQ + MBS sensor enabled the accurate and simultaneous measurement of HClO and AA with excellent selectivity and sensitivity. Measurements were highly reproducible, and the sensor was stable and exceptionally biocompatible. We successfully detected changes in the redox cycles of HClO and AA in human body fluids. This sensor is a significant advance for the investigation of reactions involved in cellular redox regulation. More importantly, we have devised a strategy for the design and construction of ratiometric electrochemical biosensors for the simultaneous determination of various bioactive species.

A highly sensitive and adjustable colorimetric assay of hydrogen sulfide by signal amplification based on G-quadruplex-Cu2+ peroxidase mimetics.

This work reports the first example of a colorimetric H2S sensor constructed through G-quadruplex-Cu2+ (G4-Cu2+) peroxidase mimetics employing Cu2+ ions and G-rich DNA with signal amplification. In the hydrogen peroxide (H2O2)-mediated oxidation of 3,3',5,5'-tetramethylbenzidine (TMB), the catalytic capacity of Cu2+ can be greatly improved in the presence of 22AG DNA, where 22AG DNA acts as a signal amplifier. However, G4-Cu2+ peroxidase mimetics lose their catalytic abilities after reacting with H2S. This is employed to develop a colorimetric assay of H2S without complex synthesis and instrumentation, with a linear range from 0.01 µM to 150 µM and a detection limit of 7.5 nM. The sensitivity of the sensor can also be adjusted by changing the concentration of Cu2+. Moreover, the developed sensor is successfully applied for the quantitative determination of H2S in human serum samples.

Photoelectrochemical aptasensor for thrombin based on Au-rGO-CuS as signal amplification elements.

A photoelectrochemical platform for thrombin determination was developed based on Au-rGO-CuS as multiple signal amplification elements. CuInS2 QDs was used to sensitize burr-shape TiO2 (b-TiO2) to obtain a strong photocurrent. Under the specific recognition between aptamer and thrombin, a sandwichlike structure was formed and the Au-rGO-CuS-labeled aptamer (S2@Au-rGO-CuS) was immobilized on the electrode surface. This induced a sharp decrease in photocurrent. The phenomenon is mainly due to the fact that CuS NPs can competitively consume the light energy and electron donor with CuInS2/b-TiO2. The rGO can increase the amount of CuS NPs and the Au NPs can accelerate charge transferring which depress the recombination of photogenerated electrons and holes in CuS to further enhance the competitive capacity of CuS. The sandwichlike structure has a steric hindrance effect. Therefore, the S2@Au-rGO-CuS has a multiple signal amplification function for thrombin determination. Under optimal conditions, the PEC aptasensor exhibited a wide linear concentration range from 0.1 pM to 10 nM with a low detection limit of 30 fM (S/N = 3) for thrombin. Besides, the designed aptasensor performed well in the assay of human serum sample, indicating good potential for the determination of thrombin in real samples. Graphical abstract.

A new voltammetric sensor and its application in pharmaceutical analysis for rutin.

A new and sensitive electrochemical sensor for rutin determination was developed based on cetyltrimethylammonium chloride (CTAC) functionalized graphene (Gr) and palladium nanoparticles (Pd) (CTAC-Gr-PdNPs) composite. Rutin displayed remarkably increased electrochemical activity on the CTAC-Gr-PdNPs composite modified electrode due to the synergistic effect of the large surface area and electrocatalytic activity of both Gr and Pd nanoparticles, which offers the feasibility for highly sensitive determination of rutin via electrochemistry. Under the optimal experimental conditions, the oxidation peak current of rutin was proportional to its concentration in the range of 2.0 × 10-8-1.0 × 10-6 mol L-1, and the limit of detection (LOD) was 5 nM (S/N = 3). The developed method was successfully applied to determine rutin in pharmaceuticals with satisfactory recoveries, which shows that the fabricated sensor has potential in pharmaceutical analysis.

A sandwich-type electrochemical aptasensor for the carcinoembryonic antigen via biocatalytic precipitation amplification and by using gold nanoparticle composites.

A sandwich-type electrochemical aptasensor is described for detecting the carcinoembryonic antigen (CEA) with high sensitivity and accuracy. Two kinds of nanomaterials are used. The first was obtained by modifying gold nanoparticles with reduced graphene oxide and hemin (Hemin-rGO-AuNPs). The second consists of horseradish peroxidase-modified organic-inorganic hybrid nanoflowers linked to gold nanoparticles to obtain an architecture of type HRP-Cu3(PO4)2-HNF-AuNPs). These serve as carriers for two aptamers (apt1 and apt2) against CEA. Simultaneously, they were used to catalyze the precipitation reaction between 4-chloro-1-naphthol(4-CN) and H2O2. A sandwich-type assay linked to enzyme inhibition amplification was established for electrochemical determination of CEA. Under optimal experimental conditions and by using differential pulse voltammetry, the response peak currents (best measured at -0.34 V vs. Ag/AgCl) increases linearly with the logarithm of the CEA concentration in the range between 100 fg mL-1 and 100 ng mL-1. The detection limit is as low as 29 fg mL-1. Graphical abstract Schematic representation of the sandwich-type electrochemical aptasensor based on signal inhibition amplification from biocatalytic precipitation reaction. (HRP-Cu3(PO4)2 hybrid nanoflowers: Horseradish Peroxidase-Cu3(PO4)2 hybrid nanoflowers; AuNPs: Gold Nanoparticles; Hemin-rGO-AuNPs: Hemin-Reduced Graphene Oxide-Gold Nanoparticles; BSA: Bovine Serum Albumin; CEA: Carcinoembryonic Antigen; CEAapt1: 5'-SH-(CH2)6-ATA CCA GCT TAT TCA ATT-3'; CEAapt2: 5'-NH2-(CH2)6-AGG GGG TGA AGG GAT ACC C-3'; GCE: Glassy carbon electrode; 4-CN: 4-Chloro-1-naphthol; DPV: Differential pulse voltammetry).

Lanthanide coordination polymer nanoparticles as a turn-on fluorescence sensing platform for simultaneous detection of histidine and cysteine.

In this work, we propose a strategy for the fluorescence assay of histidine (His) and cysteine (Cys) by using lanthanide coordination polymer nanoparticles (Ln-CPN) as a fluorescent probe. The Ln-CPN were prepared by self-assembly of adenosine monophosphate (AMP) with Tb3+, i.e. AMP-Tb. The nonluminescent AMP-Tb could be efficiently sensitized by 5-sulfosalicylic acid (SSA). The fluorescence of AMP-Tb-SSA can be remarkably quenched by Cu2+, and that of the resulting Cu/SSA/AMP-Tb can be significantly enhanced by His and Cys. Thus, a specific fluorescence "turn-on" assay for His and Cys was developed by using Cu/SSA/AMP-Tb as a sensing platform. The enhanced fluorescence intensity was proportional to the His and Cys concentration in the range from 0.2 to 150 µM and 0.5 to 200 µM, respectively, with the detection limits of 70 nM and 100 nM. Additionally, taking advantage of a masking agent for His, the differentiation of His from Cys was achieved in our system. Furthermore, the protocol can also work well for analyzing His and Cys in practical plasma samples with recovery in the range of 100.3-104.1%.

Tetraazacalix[2]arence[2]triazine Coated Fe3O4/SiO2 Magnetic Nanoparticles for Simultaneous Dispersive Solid Phase Extraction and Determination of Trace Multitarget Analytes.

To satisfy the requirement of simultaneous extraction and characterization of diverse kinds of multitarget analytes, the preparation, characterization, and application of a novel tetraazacalix[2]arene[2]triazine (TCT) coated magnetic nanoparticle (TCT MNP) adsorbent are presented in this paper. TCT assembles two benzene rings and two triazines with nitrogen cross-bridging links, which exhibits a unique structural framework and versatile recognition features based on the multiple recognition sites. These include π electron stacking, charge transfer, hydrogen bonding, and ion-exchange. TCT MNPs acted as a dispersive SPE adsorbent showing strong interaction with and adsorption of polycyclic aromatic hydrocarbons (PAHs), nitroaromatics, and heavy metal ions. The dispersive magnetic nanoparticle solid phase extraction (d-MNSPE) strategy with the simultaneous extraction and stepwise elution (SESE) procedure was designed and optimized for the five PAHs (phenanthrene, anthracene, pyrene, chrysene, and benzo(a)pyrene), six nitroaromatics (4-nitrotoluene, 2,4-dinitrotoluene, 2,4,6-trinitrotoluene, 4-nitrophenol, 2,4-dinitrophenol, and 2,4,6-trinitrophenol), and four metal ions (Cu, Zn, Mn, Cd) in aqueous samples. Due to the high stability, desirable durability, larger saturation magnetization, reuse and distinct enrichment capacity of TCT MNPs, the d-MNSPE method with the SESE strategy provided high recovery (>90%) and good precision (relative standard deviations, RSD < 10%). Coupled with the commonly used HPLC-fluorescence detection, HPLC-UV detection, and atomic absorption spectrometry, these trace probes in tap water, river water, and lake water were determined with very low detection limits, in the range of 0.09-0.15 pg mL-1 for PAHs, 6-11 pg mL-1 for nitroaromatics, and 17-53 pg mL-1 for metal ions after being enriched by the d-MNSPE. The determination of trace PAHs in urine samples from smokers and nonsmokers was successfully carried out with this method, which implied that the versatile TCT MNPs and the robust method together represent a significant potential application in the analysis of body fluids and disease markers. Such methods for accurate quantification of trace components in water are very valuable as they fulfill an unmet need in environmental and medicinal chemistry.

Electrochemical behavior of palmatine and its sensitive determination based on an electrochemically reduced L-methionine functionalized graphene oxide modified electrode.

A new and sensitive voltammetric sensor for palmatine, based on an electrochemically reduced L-methionine functionalized graphene oxide modified glassy carbon electrode (L-Met-ERGO/GCE), is reported. The electrochemical characteristics of palmatine at the proposed sensor were studied systematically and some dynamic parameters were calculated for the first time. A reasonable reaction mechanism for palmatine on the L-Met-ERGO/GCE electrode was proposed and discussed, and this could be a reference for the pharmacological action of palmatine in clinical study. Under optimized conditions, the peak current had a linear relationship with palmatine concentration in the range of 1 × 10(-7) to 5 × 10(-5) mol L(-1) with a detection limit of 6 × 10(-8) mol L(-1). Additionally, the proposed method was also used to detect palmatine in human urine samples, medicinal tablets and the Chinese herb Fibraurea recisa Pierre with satisfactory results.

Twice-walk strategy based on three-dimensional DNA walking machine driven by duplex-specific nuclease.

A twice-walk strategy based on a three-dimensional (3D) cleat-equipped DNA walking machine with a high signal amplification efficiency was investigated for ultrasensitive detection of miRNA. Impressively, addition of duplex-specific nuclease (DSN) just once drove the twice-walk strategy, making the strategy simpler. With the advantages of being simple, rapid and ultrasensitive, the biosensor offers potential for use in early clinical diagnosis.

A rapid one-step electrochemical method based on cleat-equipped molecular walking machine.

Various nucleic acid molecular machines have emerged in recent years. However, when the nucleic acid tracks are fully depleted, these walkers are highly susceptible to premature release or stalling in regions where the tracks are locally exhausted. In this work, a molecular walking machine with a cleat domain preventing dissociation from the track was explored for ultrasensitive detection of miRNA. It has been verified that the cleat design can enhance the signal amplification efficiency of molecular walking machines for electrochemical miRNA-141 detection. Notably, the single-step electrochemical biosensing platform utilizing the cleat-equipped molecular walking machine (CMWM) is exceptionally straightforward and rapid, concluding the reaction within 90 min and achieving a remarkable low detection limit of 0.26 fM. The proposed molecular walking machine with this specific cleat structure was utilized for the identification of miRNA-141 in cellular lysates, exhibiting remarkable selectivity and consistent reproducibility, showcasing its effective utility in bioanalysis. Therefore, the cleat walker developed in this study introduces an innovative method for constructing a miRNA electrochemical biosensing platform, offering new perspectives for its application in biomolecule detection and clinical disease diagnosis.

Development and application of a new 25,27-bis(L-phenylalaninemethylester-N-carbonylmethoxy)-26,28-dihydroxy-para-tert-butylcalix[4]arene stationary phase.

A 25,27-bis(L-phenylalaninemethylester-N-carbonylmethoxy)-26,28-dihydroxy- para-tert-butylcalix[4]arene-bonded silica gel stationary phase was synthesized, structurally characterized and used for LC. Its separation mechanism was studied and compared with octadecyl-bonded stationary phase, as well as our previously prepared para-tert-butylcalix[4]arene-1,2-crown-4 stationary phase. Meanwhile, the chromatographic behaviors were investigated by using polycyclic aromatic hydrocarbons, monosubstituted benzenes, anilines, phenols, Tanaka tests solutes, fluoroquinolones, and flavonoids as probes. Mechanisms involved in the chromatographic separation included hydrophobic, π-π and π-electron transfer, hydrogen bonding, and inclusion interactions. Moreover, the column was successfully employed for the analysis of the illegal additive of melamine in milk product.

Electrochemical detection of dihydromyricetin using a DNA immobilized ethylenediamine/polyglutamic modified electrode.

A novel voltammetric sensor, based on DNA immobilized on the surface of an ethylenediamine/polyglutamic (En/PGA) modified glassy carbon electrode (GCE), was constructed and used for determination of dihydromyricetin (DMY). The electrochemical behaviour of DMY at this sensor was investigated in pH 3.6 NaAc-HAc buffer solutions by cyclic voltammetry (CV) and differential pulse anodic voltammetry (DPV). The oxidation of DMY is an adsorption-controlled irreversible process. The oxidation mechanism was proposed and discussed. It was found that the modified electrode exhibited a linear voltammetric response for DMY in the range of 4.0 × 10(-8) mol L(-1) to 2 × 10(-6) mol L(-1), with a detection limit of 2 × 10(-8) mol L(-1). The method was also applied successfully to detect DMY in an ampelopsis sample with satisfactory results.

Preparation, characterization and application of a new 25,27-bis-[2-(5-methylthiadiazole)thioethoxyl]-26,28-dihydroxy-para-tert-butyl calix[4]arene stationary phase for HPLC.

A new para-tert-butylcalix[4]arene column containing thiadiazole functional groups was prepared and used for the separation of polycyclic aromatic hydrocarbons, phenolic compounds, aromatic amines, benzoic acid and its derivatives by high-performance liquid chromatography (HPLC). The effect of organic modifier content in the mobile phase on retention and selectivity of these compounds were investigated. The results indicate that the stationary phase behaves like reversed-phase packing. However, hydrogen bonding, π-π and inclusion interactions seem to be involved in the separation process. The column has been successfully employed for the analysis of clenbuterol in pork and pig casing; the limit of detection and the limit of quantitation for this method by HPLC-UV detection was 0.03 and 0.097 µg/mL, respectively; the method is demonstrated to be suitable and a competitive alternative analytical method for the determination of clenbuterol.

An ultrasensitive label-free photoelectrochemical aptasensor based on terminal deoxynucleotidyl transferase amplification and catalytic reaction of G-quadruplex/hemin.

An ultrasensitive photoelectrochemical (PEC) analysis method based on terminal deoxynucleotidyl transferase (TdT) amplification effect and G-quadruplex/hemin (G4/hemin) catalytic reaction assisted signal amplification was prepared for the determination of ampicillin. A novel Au NPs@SnIn4S8-graphene sensitized structure was used as photoactive material to attain an intense basic photocurrent. In the presence of the target, P1 obtained by strand displacement reaction was introduced into the electrode surface, and abundant G4/hemin was formed in the presence of TdT and hemin. Subsequently, G4/hemin with horseradish peroxidase-mimicking activity catalyzed the oxidation of 4-chloro-1-naphthol by H2O2 to form benzo-4-chlorohexadienone precipitation on the modified electrode surface, which severely hindered the electron transfer and effectively inhibited the photocurrent output. The detection range of the PEC aptasensor for ampicillin was 10 fM-30 nM, and the limit of detection was 4.97 fM. The aptasensor had good stability and high sensitivity, and possessed a promising application in biological analysis and environmental monitoring.

An ultrasensitive carcinoembryonic antigen electrochemical aptasensor based on 3D DNA nanoprobe and Exo III.

In this study, a highly ordered three dimensional (3D) DNA nanostructure was self-assembled by label-free DNA nanotweezers, which was used as recognized probe to interact with target. Once the target was recognized by the 3D DNA nanoprobe (3D DNT), DNA nanotweezers opened to release target analog (T1). This recognition process was proceeded in homogeneous solution, which can avoid complex electrode modification and improve reaction efficiency. Then these target analogs were captured by the signal DNA probes (E1) modified on the electrode. In the assistance of Exo III, E1 was digested and the T1 was released to participate in the next cycle to realize signal amplification. Finally, an ultrasensitive carcinoembryonic antigen (CEA) electrochemical biosensing with a detection limit of 4.88 fg mL-1 was developed.

Ratiometric fluorescence detection of ciprofloxacin using the terbium-based coordination polymers.

Herein, a facile self-assembly through adenosine monophosphate (AMP) and luminol with Tb3+ was employed to construct a dual-ligand coordinated AMP-Tb-luminol coordination polymers (CPs), which emitted the typical fluorescence of luminol. Based on the sensitization effect of ciprofloxacin (CIP) on the luminescence of Tb3+, a ratiometric sensor was fabricated using the fluorescence of luminol as an inert reference. The fluorescent intensity ratios of Tb3+ to that of luminol enhanced linearly with the CIP concentration in the range from 5 nM to 2.5 µM with a lower limit of detection of 2 nM. In addition, the proposed ratiometric fluorescent sensor exhibited high selectivity for CIP, which could also be used to detect CIP in human blood serum (HBS) with satisfactory results. To our knowledge, this is the first demonstration of using dual-ligand coordination lanthanide (Ln)-based CPs for ratio-metric CIP assay, and this straightforward strategy may open up a new platform for designing the ratio-metric sensors based on the Ln CPs.

Electrochemical oxidation behavior of colchicine on a graphene oxide-Nafion composite film modified glassy carbon electrode.

An electrochemical sensor, based on a graphene oxide (GO)-Nafion composite film modified glassy carbon electrode (GCE), was developed and used for detection of trace amounts of colchicine. Owing to the large surface area, good conductivity of GO and good affinity of Nafion, the sensor exhibited excellent electrocatalytic activity for the oxidation of colchicine, displaying a wide linear response from 5.0 × 10(-8) to 2.0 × 10(-5) mol L(-1) and a low detection limit of 1.5 × 10(-8) mol L(-1) in 0.1 mol L(-1) H(2)SO(4) solution. And this modified electrode exhibited a superior immunity from epinephrine, dopamine and ascorbic acid interference. The method was also applied successfully to detect colchicine in medicinal tablets.

Determination of caffeine content in tea based on poly(safranine T) electroactive film modified electrode.

Safranine T was electropolymerised on a glassy carbon electrode and then characterised by scanning electron microscope (SEM), X-ray diffraction (XRD) and electrochemical impedance spectroscopy (EIS). This uniform electropolymerised film was crystallisable and showed a high electrocatalytic ability towards the oxidation of caffeine. To avoid the interferences of the anions, Nafion was covered on the surface of poly(safranine T) film modified glassy carbon electrode. As a new voltammetric sensor, this modified electrode is sensitive, selective and stable to determine caffeine content in tea. The peak current increased linearly with the concentration of caffeine in the range of 3×10(-7)-1×10(-4)M, with a detection limit of 1×10(-7)M. All of these make it a useful tool for determining caffeine content in tea. What's more, it produces much less organic waste compared with other analytical techniques.

Ratiometric fluorescence sensing of glutathione by using the oxidase-mimicking activity of MnO2 nanosheet.

In this work, a facile ratiometric fluorescence sensor for GSH measurement was designed based on MnO2 nanosheet (NS), carbon dots (CDs), as well as a simple substrate o-phenylenediamine (OPD). Herein, MnO2 NS played triple essential roles in the sensing system. First, it could be reduced by GSH through a special reaction, and therefore served as GSH recognizer. Second, it played as a fluorescence nanoquencher to strongly quench the fluorescence of CDs. Third, it could directly oxidize OPD to yield a luminescent product 2, 3-diaminophenazine (DAP) via the intrinsic oxidase-like activity. It revealed that MnO2 NS could be reduced to Mn2+ in the presence of GSH. Thus its oxidase-like activity and fluorescence quenching abilities were inhibited, which then restricted the generation of DAP and recovered the fluorescence of CDs. Based on this phenomenon, a novel ratiometric fluorescence sensor for GSH determination was fabricated by measuring the ratio of fluorescent intensity of DAP to that of CDs. Besides, the constructed ratiometric fluorescent sensor, which could be facilely operated with single-wavelength excitation, exhibited high sensitivity and selectivity with a wider linear range and a lower detection limit.

Raloxifene, identified as a novel LSD1 inhibitor, suppresses the migration of renal cell carcinoma.

Aim: As an important epigenetic modulator, histone lysine-specific demethylase 1 (LSD1) has been proved to be associated with the progression of renal cell carcinoma (RCC). Discovering novel LSD1 inhibitors offers therapeutic potential for RCC treatment. Methods & Results: We identified raloxifene as a novel LSD1 inhibitor (IC50 = 2.08 µM) through small compound library screening. Molecular docking indicated raloxifene might bind LSD1 in the flavin adenine dinucleotide (FAD) binding cavity in a reversible manner. Cell viability and migration assays showed raloxifene could suppress the proliferation and migration of RCC cells bearing overexpressed LSD1. Conclusion: Our findings indicated that LSD1 might be a promising therapeutic target for RCC and that raloxifene could serve as a lead compound for further anti-RCC metastasis drug discovery.