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An electroanalytical platform capable to take and dilute the sample has been designed in order to fully integrate the different steps of the analytical process in only one device. The concept is based on the addition of glass-fiber pads for sampling and diluting to an electrochemical cell combining a paper-based working electrode with low-cost connector headers as counter and reference electrodes. In order to demonstrate the feasibility of this all-in-one platform for biosensing applications, an enzymatic sensor for glucose determination (requiring a potential as low as -0.1 V vs. gold-plated wire by using ferrocyanide as mediator) was developed. Real food samples, such as cola beverages and orange juice, have been analyzed with the bioelectroanalytical lab-on-paper platform. As a proof-of-concept, and trying to go further in the integration of steps, sucrose was successfully detected by depositing invertase in the sampling strip. This enzyme hydrolyzes sucrose into fructose and glucose, which was determined using the enzymatic biosensor. This approach opens the pathway for the development of devices applying the lab-on-paper concept, saving costs and time, and making possible to perform decentralized analysis with high accuracy., (© 2021. The Author(s).)

Flunitrazepam is one of the frequently used hypnotic drugs to incapacitate victims for sexual assault. Appropriate diagnostic tools should be available to victims regarding the growing concern about "date-rape drugs" and their adverse impact on society. Miniaturized screen-printed potentiometric sensors offer crucial point-of-care devices that alleviate this serious problem. In this study, all solid-state screen-printed potentiometric flunitrazepam sensors have been designed. The paper device was printed with silver and carbon ink. Formation of an aqueous layer in the interface between carbon-conducting material and ion-sensing membrane nevertheless poses low reproducibility in the solid-contact electrodes. Accordingly, poly(3,4-ethylenedioxythiophene) (PEDT) nano-dispersion was applied as a conducting hydrophobic polymer on the electrode surface to curb water accumulation. Conditioning of ion-sensing membrane in the vicinity of reference membrane has been considered carefully using special protocol. Electrochemical characteristics of the proposed PEDT-based sensor were calculated and compared favorably to PEDT-free one. The miniaturized device was successfully used for the determination of flunitrazepam in carbonated soft drinks, energy drink, and malt beverage. Statistical comparison between the proposed sensor and official method revealed no significant difference. Nevertheless, the proposed sensor provides simple and user-friendly diagnostic tool with less equipment for on-site determination of flunitrazepam.

The simple and reliable detection of microRNAs is of great significance for studying the biological functions, molecular diagnosis, disease treatment and targeted drug therapy of microRNA. In this study, we introduced a novel Ti 3 C 2 Tx (MXene) aerogels (denoted as MXA) composite gold nano-particles (AuNPs)-modified disposable carbon fiber paper (CFP) electrode for the label-free and sensitive detection of miRNA-155. Firstly, in the presence of MXene, graphene oxide (GO) and ethylenediamine (EDA), the 3D MXene hydrogel was formed by self-assembly method, and then adding the freeze-dried 3D MXA dropwise to CFP. Subsequently, electrodepositing AuNPs on the CFP/MXA was done to construct a 3D disposable DNA-circuit test strip with excellent interface. Under the optimum experimental conditions, the detection limit of 3D disposable DNA circuit strip for miRNA-155 was 136 aM (S/N = 3). The CFP/MXA/AuNPs (CMA) electrode also has a wide dynamic range (20 fM to 0.4 μM), with a span of 4 orders of magnitude. Notably, we also tested the practicality of the sensor in 8 clinical samples. The technological innovations in the detection and quantification of microRNA in this work may be helpful to the study new aspects of microRNA biology and the development of diagnosis., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Rod-like graphite carbon nitride@MnO 2 (R-g-C 3 N 5 @MnO 2 ) heterostructure was prepared by in situ self-anchored growth of MnO 2 nanosheet on the surface of R-g-C 3 N 5 . The synthesized R-g-C 3 N 5 @MnO 2 heterostructure as photoactive material exhibited excellent photoelectrochemical (PEC) performance, and the prepared heterostructure-aptamer probe displayed sensitive PEC response to cTnI. Therefore, the PEC method was developed to detect cTnI based on the R-g-C 3 N 5 @MnO 2 heterostructure. It was found that the linear response to cTnI was in the range 0.001-30 ng/mL under optimized conditions, and the detection limit of the proposed sensor was 0.3 pg/mL. The PEC method displays stable photocurrent response up to 8 cycles and exhibited outstanding selectivity and sensitivity. The PEC method was successfully applied to detect cTnI in serum samples. The recoveries of cTnI detection in serums reach 95.5-104%, and the relative standard deviations range from 3.20 to 4.45%., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

The ubiquity and importance of ROS and RNS in cellular signaling, disease development, and death give rise to an outstanding interest in their detection and quantification. Among the analytical techniques available, electrochemical sensors stand out for the detection of ROS/RNS due to their high sensitivity and inherent miniaturization which allows the in situ and real-time detection together with a tunable selectivity due to the different electrochemical behavior of ROS/RNS. Nanomaterial-based enzyme-free electrochemical sensors possess improved sensitivity, selectivity, stability, and unique catalytic activities. In addition, their integration in nanoelectrodes, lab-on-chips, microfluidic systems, and stretchable electrodes allow the determination of ROS/RNS in individual cells, cell organelles, or cell populations, under different experimental conditions hardly accessible using classical detection methods., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Recent research in the field of electrochemical biosensors equipped with peptides and nanomaterials have been categorized, reviewed, and critically analyzed. Indeed, using these innovative biosensors can revolutionize biomedical diagnostics in the future. Saving lives, time, and money in this field will be considered as some main benefits of this type of diagnosis. Here, these biosensors have been categorized and evaluated in four main sections. In the first section, the focus is on investigating the types of electrochemical peptide-based nanobiosensors applied to detect pathogenic microorganisms, microbial toxins, and viruses. In the second section, due to the importance of rapid diagnosis and prognosis of various cancers, the electrochemical peptide-based nanobiosensors designed to detect cancer biomarkers have been reviewed and analyzed. In the third section, the electrochemical peptide-based nanobiosensors, which were applied to detect the essential and effective biomolecules in the various diseases, and health control, including enzymes, hormones, biomarkers, and other biomolecules, have been considered. Finally, using a comprehensive analysis, all the used elements in these biosensors have been presented as conceptual diagrams that can effectively guide researchers in future developments. The essential factors in evaluating and analyzing these electrochemical peptide-based nanobiosensors such as analyte, peptide sequence, functional groups interacted between the peptide sequences and other biosensing components, the applied nanomaterials, diagnostic techniques, detection range, and limit of detection have also been included. Other analyzable items such as the type of used redox marker and the location of the peptide sequence against the signal transducer were also considered., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

3D-printed portable devices have immense and proven potential to transform the field of electrochemiluminescence (ECL) for diverse biochemical applications. 3D printing (3DP) offers unparalleled ability to build tiny devices in a single step with high accuracy and compatibility, and integrability as per the requirement. In this study, for the first time, a six-well 3D-printed closed bipolar electrochemiluminescence (3DP-CBPE-ECL) device has been successfully fabricated and validated by performing single-step detection of various biochemicals such as glucose and choline. Luminol/H 2 O 2 -based enzymatic reactions were performed with optimized parameters for selective sensing of glucose and choline. The single-step detection of glucose and choline was accomplished for the linear ranges of 0.1 to 10 mM and 0.1 to 5 mM, with a limit of detections (LODs) of 24 µM and 10 µM, respectively. A smartphone was leveraged to execute multiple activities such as powering the ECL device, capturing ECL images, and calculating the ECL intensity of the obtained ECL signal. The feasibility of a six-well 3DP-CBPE-ECL device was tested by sensing glucose and choline simultaneously in a single device at three different concentrations. Furthermore, the concentration of glucose and choline was calculated in real blood serum using the conventional additive (spiking) method, demonstrating the high practicability of the fabricated ECL device and yielding promising findings. Finally, based on the obtained results and other advantages such as low-cost, fast prototyping and requirement of a minimum sample volume, the fabricated six-well 3DP-CBPE-ECL device has shown potential to be used in the field of biochemical applications., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Development of MXene (Ti 3 C 2 Cl 2 )-based sensing platforms by exploiting their inherent active electrochemistry is highly challenging due to their characteristic poor stability in air and water. Herein, we report a cost-effective methodology to deposit MXene on a conductive graphitic pencil electrode (GPE). MXenes can provide active surface area due to their clever morphology of accordion-like sheets; however, the disposition to stack together limits their potential applications. A task-specific ionic liquid (1-methyl imidazolium acetate) is utilized as a multiplex host material to engineer MXene interface via π-π interactions as well as to act as a selective binding site for biomolecules. The resulting IL-MXene/GPE interface proved to be a highly stable interface owing to good interactions between MXene and IL that inhibited electrode leaching and boosted electron transfer at the electrode-electrolyte interface. It resulted in robust dopamine (DA) oxidation with amplified faradaic response and enhanced sensitivity (9.61 µA µM -1  cm -2 ) for DA detection. This fabricated sensor demonstrated large linear range (10 µM - 2000 µM), low detection limit (702 nM), high reproducibility, and good selectivity. We anticipate that such platform will pave the way for the development of stable and economically viable MXene-based sensors without sacrificing their inherent properties. Scheme 1 Schematic illustration of the IL-MXene/GPE fabrication and oxidative process towards non-enzymatic dopamine sensor., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Noninvasive diagnosis using salivary samples to detect thiocyanate provides vital information on individual health. This article demonstrates the first example of a wearable sensing device to noninvasively assess thiocyanate levels. The customized screen-printed electrode system is integrated into a form of a mouthguard squarewave-voltammetric sensor toward the convenient and fast detection of the salivary biomarker within 15 s. The sensor with a protective film to mitigate the effect of biofouling offers high sensitivity and selectivity toward the detection of thiocyanate ions. Partial least square regression is applied to analyze the high-order squarewave-voltammetric data over the applied potential range of 0-1.75 V vs Ag/AgCl and quantify the thiocyanate concentration in a complex matrix. The mouthguard sensor operating under physiological conditions can monitor a wide range of thiocyanate (up to 11 mM) with a low detection limit of 30 µM. The demonstration introduces a unique approach, that obviates the requirement for blood sampling, to study thiocyanate levels of healthy people, cigarette smokers, or people with other health conditions. It is envisioned that the new cavitas device possesses a substantial promise for diverse biomedical diagnosis applications., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

For the first time, a comprehensive review is presented on the quantitative determination of narrow therapeutic index drugs (NTIDs) by nano optical and electrochemical sensors and biosensors. NTIDs have a narrow index between their effective doses and those at which they produce adverse toxic effects. Therefore, accurate determination of these drugs is very important for clinicians to provide a clear judgment about drug therapy for patients. Routine analytical techniques have limitations such as being expensive, laborious, and time-consuming, and need a skilled user and therefore  the nano/(bio)sensing technology leads to high interest., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

A novel biosensing interface for tumor markers was designed based on the atom transfer radical polymerization (ATRP) of poly(isopropenylphenol) (PPPL) in situ initiated by the fixing of p-chloromethyl benzoic acid on the surface of amino-modified electrodes. It was found that the electrochemical activity of PPPL itself can provide sufficient signals for these biosensors, which can avoid signal leakage and streamline the interface modification process. Cu(II) ions absorbed on the carbon spheres and then were released via acid stimulation to act as a catalyst to participate in the interface polymerization with ATRP. As the concentration of targets increased, more Cu(II) ions were released, and the electrochemical signal of polymers was enhanced. Therefore, the sensitive detection of carbohydrate antigen 19-9 (CA19-9) as a model target was achieved, with an ultralow limit of detection of 39 µU mL -1 and wide detection range from 100 µU mL -1 to 100 U mL -1 under optimal conditions. Furthermore, this method achieved satisfying performance in human blood serum with good inter-assay precision (RSD < 6%) and satisfactory recovery of ~ 99-105%. According to the results, this work is of great significance for constructing biosensor interfaces via in situ polymerization. A novel biosensing interface for tumor marker was designed based on atom transfer radical polymerization (ATRP), which poly(isopropenylphenol) with electrochemical signal was fabricated in situ on electrode., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

A novel electrochemical DNA sensor was designed to detect Helicobacter pylori based on accordion-like Ti 3 C 2 T x . Here the multilayer Ti 3 C 2 T x obtained by DMSO delamination was used to modify the glass carbon electrode, with a large specific surface area and excellent conductivity. Au nanoparticles were supported on the modified electrode and worked as an effective carrier to fix the capture probe (cpDNA) with sulfhydryl group through the firm binding of Au-S bond. Such an accordion-like Ti 3 C 2 T x structure provides an ultrahigh electroactive surface area and ample binding sites for accommodating Au nanoparticles, which is advantageous for the signal amplification during the detection. And further, the sandwich structure formed by hybridizing cpDNA with target DNA sequence (tDNA) and rpDNA (rpDNA is a strand of DNA that can be base-paired with the tested tDNA) increases greatly the current signal and enhances the sensitivity of the electrochemical DNA sensor. Under optimal conditions, the developed electrochemical DNA sensor showed a wide linear range from 10 -11 to 10 -14  M and a low detection limit of 1.6 × 10 -16  M and exhibited good sensitivity, reproducibility, and stability. A novel electrochemical DNA sensor with simple sandwich structure was designed to detect H. pylori based on accordion-like Ti 3 C 2 T x .

A novel immunoassay is introduced based on co-reactant enhancing strategy for the electrochemiluminescent (ECL) determination of CA15-3 and CA72-4 tumor markers in real samples. For the preparation of the signaling probe, CA15-3 and CA72-4 antibodies first were labeled using Ru(bpy) 3 2+ -N-hydroxysuccinimide ester (Ru(bpy) 3 2+ -NHS) and conjugated with L-cysteine capped cadmium selenide (CdSe) quantum dots. Finally, it was cross-linked with chitosan-grafted graphene oxide (GO@CS) nanocomposite. The capture probe was constructed by deposition of multi-walled carbon nanotubes (MWCNT) at the surface of dual-working gold screen-printed electrodes (MWCNT-dwSPE) and covalent attachment of capture CA15-3 and CA72-4 antibodies to MWCNT-dwSPE. ECL signals were recorded by applying cyclic potential ranging from 0.3 to 1.1 V (vs. pseudo-reference Ag/AgCl) at the scan rate of 100 mV.s -1 . This immunoassay was used for determination of CA15-3 and CA72-4 in real samples the detection limits of 9.2 μU.ml -1 and 89 μU.ml -1 within linear ranges of 10 μU.ml -1 -500 U.ml -1 and 100 μU.ml -1 -150 U.ml -1 , respectively. This immunoassay also showed acceptable accuracy with recoveries in the range 96.5-108 % and high reproducibility with RSD of 3.1 and 4.9.

Since the COVID-19 disease caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS- CoV-2) was declared a pandemic, it has spread rapidly, causing one of the most serious outbreaks in the last century. Reliable and rapid diagnostic tests for COVID-19 are crucial to control and manage the outbreak. Here, a label-free square wave voltammetry-based biosensing platform for the detection of SARS-CoV-2 in nasopharyngeal samples is reported. The sensor was constructed on screen-printed carbon electrodes coated with gold nanoparticles. The electrodes were functionalized using 11-mercaptoundecanoic acid (MUA) which was used for the immobilization of an antibody against SARS-CoV-2 nucleocapsid protein (N protein). The binding of the immunosensor with the N protein caused a change in the electrochemical signal. The detection was realised by measuring the change in reduction peak current of a redox couple using square wave voltammetry at 0.04 V versus Ag ref. electrode on the immunosensor upon binding with the N protein. The electrochemical immunosensor showed high sensitivity with a linear range from 1.0 pg.mL -1 to 100 ng.mL -1 and a limit of detection of 0.4 pg.mL -1 for the N protein in PBS buffer pH 7.4. Moreover, the immunosensor did not exhibit significant response with other viruses such as HCoV, MERS-CoV, Flu A and Flu B, indicating the high selectivity of the sensor for SARS-CoV-2. However, cross reactivity of the biosensor with SARS-CoV is indicated, which gives ability of the sensor to detect both SARS-CoV and SARS-CoV-2. The biosensor was successfully applied to detect the SARS-CoV-2 virus in clinical samples showing good correlation between the biosensor response and the RT-PCR cycle threshold values. We believe that the capability of miniaturization, low-cost and fast response of the proposed label-free electrochemical immunosensor will facilitate the point-of-care diagnosis of COVID 19 and help prevent further spread of infection.

Folic acid (FA) is essential for human health, particularly for pregnant women and infants. In this work, a glassy carbon electrode (GCE) was modified by a bimetallic layer of Cu/Co nanoparticles (CuNPs/CoNPs) as a synergistic amplification element by simple step-by-step electrodeposition, and was used for sensitive detection of FA. The proposed CuNPs/CoNPs/GCE sensor was characterized by differential pulse voltammetry (DPV), electrochemical impedance spectroscopy (EIS) and field emission scanning electron microscopy (FESEM). Then, under optimal conditions, a linear relationship was obtained in the wide range of 110.00-1750.00 μM for the detection of FA with a limit of detection (LOD) of 34.79 μM (S/N = 3). The sensitivity was calculated as 0.096 μA μM -1 cm -2 . Some interfering compounds including glucose (Glc), biotin, dopamine (DA), and glutamic acid (Glu) showed little effect on the detection of FA by amperometry (i-t). Finally, the average recovery obtained was in a range of 91.77-110.06%, with a relative standard deviation (RSD) less than 8.00% in FA tablets, indicating that the proposed sensor can accurately and effectively detect the FA content in FA tablets., (© 2022. Springer-Verlag GmbH Germany, part of Springer Nature.)

Monitoring of cerebrospinal fluid (CSF) microRNAs (miRs) offers a promising option for the diagnosis and management of patients with central nervous system tumors. However, the sensitive detection of miRs in clinical CSF samples has been hindered by the ultra-low abundance of target miRs. Here, we report an electrochemical biosensor for the highly sensitive label-free detection of CSF miR-21 relying on target-induced redox signal amplification (eTIRSA). The biosensor was developed by covalently assembling the capture stands partially complementary to miR-21 on the gold nanoparticle-coated glassy carbon electrode. In the presence of miR-21, the short capture stand hybridized with the partial bases of miR-21, allowing the rest sequence of the target molecule to further bind with a long guanine-rich sequence which could specifically adsorb a number of methylene blue indicators, thus generating an amplified electrochemical redox signal, typically at a working potential of - 0.19 V (vs. SCE). The response of the surface-bound methylene blue indicators was positively correlated to the concentration of miR-21, providing a dynamic range of 0.5-80 pM and a limit of detection down to 56 fM. Moreover, the eTIRSA biosensor had high specificity with single-base resolution and exhibited good performance for label-free quantification of miR-21 in medulloblastoma cell extracts and clinical CSF samples and for accurate discrimination of medulloblastoma against non-cancer controls, indicating its potential application in CSF miR-based liquid biopsy of brain cancers., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Alzheimer's disease, as a most prevalent type of dementia, is quickly becoming one of the most expensive, lethal, and burdening diseases of this century. Though there are still no efficient therapies, early diagnosis and intervention are important directive significance to clinical works. Here, we develop a portable electrochemical micro-workstation platform consisting of an electrochemical micro-workstation and integrated electrochemical microarray for simultaneously detecting multiple AD biomarkers including Aβ40, Aβ42, T-tau, and P-tau181 in serum. The integrated electrochemical microarray is mainly used for droplet sample manipulation and signal generation. The micro-workstation can regulate signals and transfer the signals to a smartphone by Bluetooth embedded inside. This portable electrochemical micro-workstation platform exhibits excellent analysis performance. The LODs for Aβ40, Aβ42, T-tau, and P-tau181 are 0.125 pg/mL, 0.089 pg/mL, 0.142 pg/mL, and 0.176 pg/mL, respectively, which satisfies the needs of detecting AD biomarkers in serum. The combination of portable micro-workstation and integrated electrochemical microarray provides a promising strategy for the early diagnosis of Alzheimer's disease and personal healthcare., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Rapid and selective sensing of KRAS gene mutation which plays a crucial role in the development of colorectal, pancreatic, and lung cancers is of great significance in the early diagnosis of cancers. In the current study, we developed a simple electrochemical biosensor by differential pulse voltammetry technique for the specific detection of KRAS mutation that uses the mismatch-specific cleavage activity of T7-Endonuclease I (T7EI) coupled with horseradish peroxidase (HRP) to catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) substrate in the presence of hydrogen peroxide (H 2 O 2 ). In addition, we synthesized the nanocomposite composed of multi-walled carbon nanotube/chitosan-ionic liquid/gold nanoparticles (MWCNT/Chit-IL/AuNPs) on screen-printed carbon electrode surface to increase the electrode surface area and electrochemical signal. In principle, T7E1 enzyme recognized and cleaved the mismatched site formed by the presence of KRAS gene mutation, removing 5'-biotin of capture probes and subsequently reducing the differential pulse voltammetry signal compared to wild-type KRAS gene. With this proposed strategy, a limit of detection of 11.89 fM was achieved with a broad linear relationship from 100 fM to 1 µM and discriminated 0.1% of mutant genes from the wild-type target genes. This confirms that the developed biosensor is a potential platform for the detection of mutations in early disease diagnosis., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Wearable film-based smart biosensors have been developed for real-time biomolecules detection. Particularly, interfacial co-assembly of reduced graphene oxide-prussian blue (PB-RGO) film through electrostatic interaction has been systematically studied by controllable pH values, achieving optimal PB-RGO nanofilms at oil/water (O/W) phase interface driven by minimization of interfacial free energy for wearable biosensors. As a result, as-prepared wearable biosensors of PB-RGO film could be easily woven into fabrics, exhibiting excellent glucose sensing performance in amperometric detection with a sensitivity of 27.78 µA mM -1  cm -2 and a detection limit of 7.94 μM, as well as impressive mechanical robustness of continuously undergoing thousands of bending or twist. Moreover, integrated wearable smartsensing system could realize remotely real-time detection of biomarkers in actual samples of beverages or human sweat via cellphones. Prospectively, interfacial co-assembly engineering driven by pH-induced electrostatic interaction would provide a simple and efficient approach for acquiring functional graphene composites films, and further fabricate wearable smartsensing devices in health monitoring fields., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

In this study, Lba Cas12a (Cpf1) as one of the CRISPR systems from Lachnospiraceae bacterium was coupled with a hybridization chain reaction (HCR) to develop an electrochemical biosensor for detecting the pathogenic bacterium, Salmonella typhimurium. Autonomous cross-opening of functional DNA hairpin structures of HCR yielded polymer double-stranded DNA wires consisting of numerous single-stranded DNAs, which initiated the trans-cleavage activity of CRISPR-Cas12a to indiscriminately cleave random single-stranded DNA labeling electrochemical tags on the surface of the electrode. It led to a variation in the electron transfer of electrochemical tags. The polymer double-stranded DNA of HCR was immobilized on dynabeads (DBs) via the S. typhimurium aptamer and released from DBs. The established method could selectively and sensitively quantify S. typhimurium in samples with detection limits of 20 CFU/mL. Our study provides a novel insight for exploring universal analytical methods for pathogenic bacteria based on CRISPR-Cas12a coupled with HCR., (© 2021. Springer-Verlag GmbH Germany, part of Springer Nature.)

Microfluidic devices can provide a versatile, cost-effective platform for disease diagnostics and risk assessment by quantifying biomarkers. In particular, simultaneous testing of several biomarkers can be powerful. Here, we critically review work from the previous 4 years up to February 2021 on developing microfluidic devices for multiplexed detection of biomarkers from samples. We focus on two principal approaches: electrical and optical detection methods that can distinguish and quantify biomarkers. Both electrical and spectroscopic multiplexed detection strategies are being employed to reach limits of detection below clinical sample levels. Some of the most promising strategies for point-of-care assays involve inexpensive materials such as paper-based microfluidic devices, or portable and accessible detectors such as smartphones. This review does not comprehensively cover all multiplexed microfluidic biomarker studies, but rather provides a critical evaluation of key work and suggests promising prospects for future advancement in this field. Electrical and optical multiplexing are powerful approaches for microfluidic biomarker analysis., (© 2021. Springer-Verlag GmbH Germany, part of Springer Nature.)

Abrin is a highly toxic ribosome-inactivating protein, which could be used as a biological warfare agent and terrorist weapon, and thus needs to be detected efficiently and accurately. Affibodies are a new class of engineered affinity proteins with small size, high affinity, high stability, favorable folding and good robustness, but they have rarely played a role in biological detection. In this work, we establish a novel electrochemiluminescence (ECL) method for abrin detection with a phage display affibody as the specific probe for the first time, to our knowledge, and a portable biosensor based on a screen-printed electrode (SPE) as the testing platform. On the basis of the double antibody sandwich structure in our previous work, we used a phage display affibody instead of monoclonal antibody as a new specific labeled probe. Due to numerous signal molecules labeled on M13 phages, significant signal amplification was achieved in this experiment. Under optimized conditions, a linear dependence was observed from 0.005 to 100 ng/mL with a limit of detection (LOD) of 5 pg/mL. This assay also showed good reproducibility and specificity, and performed well in the detection of simulated samples. Considering its high sensitivity, interference resistance and convenience, this new biosensing system based on phage display affibodies and a portable ECL biosensor holds promise for in situ detection of toxins and pollutants in different environments., (© 2021. Springer-Verlag GmbH Germany, part of Springer Nature.)

A sensitive detection of carbohydrate antigen 15-3 (CA15-3) levels may allow for early diagnosis and monitoring the treatment of breast cancer, but this can only be made in routine clinical practice if low-cost immunosensors are available. In this work, we developed a sandwich-type electrochemical immunosensor capable of rapid detection of CA15-3 with an ultra-low limit of detection (LOD) of 0.08 fg mL -1 within a wide linear concentration range from 0.1 fg mL -1 to 1 µg mL -1 . The immunosensor had a matrix of a layer-by-layer film of Au nanoparticles and reduced graphene oxide (Au-rGO) co-electrodeposited on screen-printed carbon electrodes (SPCE). The high sensitivity was achieved by using secondary antibodies (Ab 2 ) labeled with horseradish peroxidase (HRP) in the presence of hydrogen peroxide (H 2 O 2 ) as signal amplifiers, and hydroquinone (HQ) was used as an electron mediator. The immunosensor was selective for CA15-3 in human serum and artificial saliva samples, robust, and stable to permit storage at 4 °C for more than 30 days. With its high performance, the immunosensor may be incorporated into future point-of-care (POC) devices to determine CA15-3 in distinct biological fluids, including in blood and saliva samples., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

A novel electrochemical sensor has been fabricated for sensitive determination of zearalenone (ZEA) in food samples based on molybdenum disulfide quantum dots (MoS 2 QDs) and two-dimensional titanium carbide (2D-Ti 3 C 2 T x MXene) co-modified multi-walled carbon nanotube (MWCNT) heterostructure. Physical and electrochemical characterizations reveal that 2D-Ti 3 C 2 T x and MoS 2 QDs co-modified MWCNTs yields synergistic signal amplification effect, together with large specific surface area and excellent conductivity for the heterostructure, endowing the developed sensor with high detection performance to ZEA. Under optimized conditions, the sensor shows a wide linear range from 3.00 to 300 ng mL -1 and a low limit of detection (LOD) of 0.32 ng mL -1 , which is far lower than the maximum residue limits (MRLs) settled by the European Commission. In addition, it exhibits excellent selectivity, high reproducibility with a relative standard deviation (RSD) of 1.1%, and good repeatability (RSD 1.1%). In real sample analysis recoveries ranged from 94.8 to 105% showing the proposed electrochemical sensor has high potential in practical applications. This work presents an effective and valuable pathway for the use of novel heterostructure in the biosensing field., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

As well known, the electrochemiluminescence (ECL) of tris(2,2'-bipyridine)ruthenium(II) (Ru(bpy) 3 2+ ) heavily relies on highly positive or negative triggered voltage, prejudicing the detection toward the bio-molecules. In this work, Ru(bpy) 3 2+ could generate enhanced and stable ECL at a low potential of 0.05 V (vs. Ag/AgCl) on graphene-PtPd hybrid, attributing to its excellent electrocatalysis from the synergistic effect between Pt and Pd. The obtained low-potential-driven ECL could be quenched by MoS 2 nanoflowers. Based on the quenching effect, a sandwich "signal-off" ECL immunosensor was fabricated to sensitively detect carcinoembryonic antigen (CEA). A linear calibration curve from 1 fg mL -1 to 1 ng mL -1 was obtained along with a low detection limit of 0.54 fg mL -1 (S/N = 3) under optimal conditions. The sensor showed satisfactory specificity, stability, and reproducibility and was successfully applied to determine CEA in actual samples. The recoveries ranged from 98.80 to 100.23%, and the relative standard deviation (RSD) was lower than 5%. Above all, this work explored new materials in low-potential-driven ECL system and provided a reliable sensing strategy for clinical applications., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

This research proposed a replacement-type electrochemiluminescent (ECL) aptasensor for lysozyme (LYZ) detection at trace levels based on a full-electric modification electrode (FEMG) coupled to silica-coated Ru(bpy) 3 2+ /silver nanospheres (Ru/SNs@SiO 2 ). The multi-walled carbon nanotubes-doped-thionine (MWCNTs/PTn) electropolymerized modified electrode was decorated with electrodeposited gold nanoparticles (GNs) to form the FEMG. Then, the FEMG was utilized as sensing substrates for the immobilization of the anti-lysozyme aptamer (LA); the stability and number of LA attaching onto the FEMG were dramatically increased. The ECL measurement was used to evaluate the hybridization reaction of LA and the Ru/SNs@SiO 2 marked DNA probe, and it was noted as I a . After the combination of the LA with the LYZ, the target-triggered replacement of the DNA probe was actualized and the ECL measurement descended to I b . The ECL difference (ΔI ECL  = I a  - I b ) before and after the replacement event was utilized for quantitation of LYZ. As a result, the fabricated aptasensor with great sensitivity and specificity achieved a wide linear range (10 fM-10 pM) and a low limit of detection (5 fM). It obtained satisfactory recovery for the detection of LYZ in human serum, and the results were identified with the LYZ ELISA kit. Therefore, the proposed ECL sensor is expected to become a promising approach in the field of biomolecule detection., (© 2021. Springer-Verlag GmbH Germany, part of Springer Nature.)

Peroxynitrite anion (ONOO - ) is an important in vivo oxidative stress biomarker whose aberrant levels have pathophysiological implications. In this study, an electrochemical sensor for ONOO - detection was developed based on graphene nanoplatelets-cerium oxide nanocomposite (GNP-CeO 2 ) incorporated polyaniline (PANI) conducting hydrogels. The nanocomposite-hydrogel platform exhibited distinct synergistic advantages in terms of large electroactive surface coverage and providing a conductive pathway for electron transfer. Besides, the 3D porous structure of hydrogel integrated the GNP-CeO 2 nanocomposite to provide hybrid materials for the evolution of catalytic activity towards electrochemical oxidation of ONOO - . Various microscopic and spectroscopic characterization techniques endorsed the successful formation of GNP-CeO 2 -PANI hydrogel. Cyclic voltammetry (CV) measurements of GNP-CeO 2 -PANI hydrogel modified screen-printed electrodes (SPE) were carried out to record the current changes influenced by ONOO - . The prepared sensor demonstrated a significant dose-dependent increase in CV peak current within a linear range of 5-100 µM (at a potential of 1.12 V), and a detection limit of 0.14 with a sensitivity of 29.35 ± 1.4 μA μM -1 . Further, a customized microfluidic flow system was integrated with the GNP-CeO 2 -PANI hydrogel modified SPE to enable continuous electrochemical detection of ONOO - at low sample volumes. The developed microfluidic electrochemical device demonstrated an excellent sensitivity towards ONOO - under optimal experimental conditions. Overall, the fabricated microfluidic device with hybrid hydrogels as electrochemical interfaces provides a reliable assessment of ONOO - levels. This work offers considerable potential for understanding the oxidative stress-related disease mechanisms through determination of ONOO - in biological samples., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Advances in materials science have accelerated the development of diagnostic tools with the last decade witnessing the development of enzyme-free sensors, owing to the improved stability, low cost and simple fabrication of component materials. However, the specificity of non-enzymatic sensors for certain analytes still represents a challenging task, for example the determination of cholesterol level in blood is vital due to its medical relevance. In this work, a reagent displacement assay for cholesterol sensing in serum samples was developed. It is based on coating of a glassy carbon electrode with a polymer of intrinsic microporosity (PIM) that forms a host-guest complex with methylene blue (MB). In the presence of cholesterol, the MB electroactive probe was displaced due to the stronger association of cholesterol guest to the PIM host. The decrease in the oxidative current was proportional to the cholesterol concentration achieving a detection limit of approximately 0.1 nM. Moreover, to further assist the experimental studies, comprehensive theoretical calculations are also performed by using density functional theory (DFT) calculations., (© 2021. Springer-Verlag GmbH Germany, part of Springer Nature.)

A hydrogel based on titanium dioxide/MXene with polyvinyl alcohol/graphene oxide (TiO 2 /MXene-PVA/GO) composite was successfully formulated and applied to modify a screen-printed carbon electrode (SPCE) for urinary norepinephrine (NE) detection. The characterization confirmed that a nanocomposite hydrogel structure of TiO 2 /MXene-PVA/GO was formed. The as-prepared hydrogel substantially enhanced the sensor performances due to electrocatalytic activity of TiO 2 , high conductivity of MXene, and auto-sample preconcentration via PVA/GO hydrogel. The electrochemical behavior of NE was investigated by cyclic voltammetry and amperometry. Under optimized conditions, the TiO 2 /MXene-PVA/GO hydrogel/SPCE response due to the oxidation of NE at +0.4 V (vs. Ag|AgCl) is proportional to the concentration of NE over 0.01 to 1.00 μM (R 2  = 0.9968) and 1.00 to 60.0 μM (R 2  = 0.9936) ranges with a detection limit (3σ) of 6 nM without interferent effect from common interferences in urine. Furthermore, this sensor was employed for urinary NE determination and validated by high performance liquid chromatography (HPLC) with a UV detector at 280 nm; the average recovery was found to be 97.6 to 102%, with a relative standard deviation (RSD) less than 4.9%. This device was sensitive enough to evaluate an early stage of neurological disorder via detecting clinically relevant NE level. Eventually, it was integrated with pantyliners which could be a potential wearable sensor in the near future., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

A highly sensitive electrochemical biosensors has been developed for the detection of multiplex micro ribonucleic acids (miRNAs) by modifying an electrode with reduced graphene oxide/poly(2-aminobenzylamine)/gold nanoparticles and adopting porous, hollow silver-gold nanoparticles as tagged labeling with metal ions. In addition, an anti-deoxyribonucleic acid (DNA)-RNA hybrid [S9.6] antibody was used to detect different hybridized capture DNAs and miRNAs that can detect multiple miRNAs simultaneously. The developed electrochemical platform exhibits high selectivity, stability, and sensitivity with a wide linear range from 1 fM to 10 nM and a low detection limit of 0.98 fM, 3.58 fM, and 0.25 fM for miRNA-155, miRNA-21, and miRNA-16, respectively. In addition, the proposed electrochemical biosensor capable for the simultaneous detection of miRNA-155, miRNA-16, and miRNA-21, which are breast cancer biomarkers, in normal human serum, can be adopted and potentially used for breast cancer screening., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Three-dimensional flowerlike Au@(MoS 2 /GO/o-MWNTs) nanohybrids (abbreviated as AMGMs) were synthesized and then introduced into an electrochemiluminescence (ECL) system as a new co-reaction accelerator for the ultrasensitive prostate-specific antigen (PSA). The AMGMs not only served as a substrate with good conductivity and a large specific surface area for loading abundant primary antibodies but also acted as an effective co-reaction accelerator; the co-reaction accelerator could interact with a co-reactant rather than the luminophore to boost the generation of free radical intermediates, thereby producing abundant excited states of luminophores to amplify the ECL signal response. Additionally, an anticipated signal amplification strategy based on the hybridization chain reaction (HCR) was developed by gathering a large amount of a DNA initiator on gold nanoparticles. These gathered DNA initiators could generate multiple DNA concatemers and attach more signal molecules, which resulted in outstanding exponential signal amplification. Consequently, the ECL immunosensor demonstrated high sensitivity, with a linear range from 0.1 pg mL -1 to 50 ng mL -1 and a detection limit of 0.028 pg mL -1 . In addition, the immunosensor displayed excellent stability and selectivity. It was evaluated by analyzing human serum sample. The recovery obtained was 98.80-112.00% and the RSD was 1.73-3.12%, indicating the immunosensor could be applied to the simultaneous detection of PSA in human serum samples. Graphical abstract., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Tetrabromobisphenol A (TBBPA) is a kind of brominated flame retardant that is usually added to products to reduce their flame retardancy. However, its extensive use has resulted in their residues being found in the environment, which is very harmful. Herein, an indirect competitive immunosensor has been established for TBBPA detection based on the signal amplification system. Pd nanospheres in situ reduced on the surface of MnO 2 nanosheet hybrid (MnO 2 /Pd) was used as the label for the secondary antibody through the Pd-N bond, and gold-toluidine blue composite was loaded onto MWCNTs (MWCNTs/Au-TB), which functioned as the platform for the immunosensor. The spherical structure of Pd had abundant catalytic active sites, which enhanced the catalytic activity of MnO 2 /Pd as the label, hence amplifying the signal response. Besides, MWCNTs/Au-TB improved electron transfer and produced a strong signaling pathway for immobilizing antigens through the Au-NH 2 bond, which can specifically recognize primary antibodies to improve sensitivity. The immunosensor had a linear concentration range of 0-81 ng/mL, a low detection limit of 0.17 ng/mL (S/N = 3), with good stability, selectivity, and reproducibility based on the above advantages. Additionally, the acceptable accuracy and recoveries (recoveries, 92-124%; CV, 3.3-8.8%) in the real water sample analysis indicated that this strategy is promising for emerging pollutant analysis.

Electrochemiluminescence (ECL) can be produced through two main routes: annihilation route and coreactant route. The vast majority of applications of ECL are based on coreactant ECL which can be generated in aqueous media at relatively low potentials compared with organic solvents. However, the development of more efficient ECL systems remains a compelling goal. Co-reaction accelerator (CRA) can significantly enhance the ECL signal through promoting more production of the coreactant intermediate. Compared with other ECL enhancement strategies, the CRA protocol is distinctive owing to its diverse, simple, and highly effective features. Various species such as inorganic compound, organic compound, and nanomaterials (NMs) have been developed as CRA and NM CRA has gained particular attention owing to their unique properties of excellent catalytic behavior and large surface area. By integration with the inherent advantages of ECL, bioanalysis based on CRA-enhanced ECL showed excellent performance such as ultrahigh sensitivity, wide dynamic range, low cost, simple instrumentation, and measurements in complex media. It has been extensively applied in various fields including clinical diagnosis, environmental monitoring, and food safety. Therefore, it is of great interest to present a systematic and critical review on the advances in ECL CRA. Herein, the recent progress on CRA and its applications in ECL bioanalysis are summarized by illustrating some representative work and a discussion of the future development trends of CRA ECL is offered.

The integration of large surface area and high catalytic profiles of Cu-MOF and CuO nanoparticles is described toward electrochemical sensing of nitric oxide (NO) in a microfluidic platform. The CuO/Cu-MOF nanocomposite was prepared through hydrothermal method, and its formation was confirmed by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and energy-dispersive X-ray spectroscopy (EDS). The CuO/Cu-MOF nanostructured modified Au electrodes enabled electrocatalytic NO oxidation at 0.6 V vs. reference electrode, demonstrating linear response over a broad concentration range of 0.03-1 μM and 1-500 μM with a detection limit of 7.8 nM. The interference effect of organic molecules and common ions was negligible, and the sensing system demonstrated excellent stability. Finally, an electrochemical microfluidic NO sensor was developed to detect of NO released from cancer cells, which were stimulated by L-arginine. Furthermore, in the presence of Fe 3+ , the stressed cells produced more NO. This work offers considerable potential for its practical applications in clinical diagnostics through determination of chemical symptoms in microliter-volume biological samples. Electrochemical microfluidic NO sensor was developed for detection of NO released from cancer cells. This miniaturized device consumes less materials and provides the basis for greener analytical chemistry.

Entomotoxicology allows the detection and analysis of substances such as poisons, drugs, and metals in necrophagous insects using analytical protocols. In a forensic situation related to death by gunshot, the gunshot residue (GSR) is dispersed at the crime scene and may be consumed by necrophagous insects. Lead (Pb) is the most abundant metal in GSR samples and it can be determined using non-portable methods. However, the toxicity effects of GSR samples on Lucilia cuprina (Diptera: Calliphoridae) and the detection of Pb via portable electrochemical methods have not been investigated. This study describes for the first time the toxicity analysis of Pb on immature L. cuprina through their survival rate and influence of Pb on immature development. In addition, the bioaccumulation of Pb in the larvae samples was determined based on square wave anodic stripping voltammetry (SWASV) measurements. The results revealed a low limit of detection to Pb (6.5 μg L -1 ) and the analytical performance was satisfactory because it measures Pb levels in larvae exposed to a diet containing 50 μg Pb g -1 . Furthermore, the levels of Pb influenced the survival rate and development time of the immature L. cuprina. Larvae exposed to a high concentration of the metal (50 μg Pb g -1 ) showed statistically significant changes (p < 0.05). The presence of Pb in immature L. cuprina can be used to estimate the post-mortem interval; thus, the present study provides important information in forensic entomology.

In recent years, an increasing number of studies has demonstrated that redox polymers can be used in simple and effective electrochemical sensing platforms due to their fast electron transfer and electrocatalytic ability. To develop more sensitive and selective electrochemical (bio)sensors, the electrocatalytic properties of redox polymers and the electrical, mechanical, and catalytic properties of various nanomaterials are combined. This review aims to summarize and contribute to the development of (bio)sensors based on polyphenazine or polytriphenylmethane redox polymers combined with nanomaterials, including carbon-based nanomaterials, metal/metal oxide, and semiconductor nanoparticles. The synthesis, preparation, and modification of these nanocomposites is presented and the contribution of each material to the performance of (bio)sensor has been be examined. It is explained how the combined use of these redox polymers and nanomaterials as a sensing platform leads to improved analytical performance of the (bio)sensors. Finally, the analytical performance characteristics and practical applications of polyphenazine and polytriphenylmethane redox polymer/nanomaterial-based electrochemical (bio)sensors are compared and discussed.

A sensitive photoelectrochemical (PEC) sensor based on hexagonal carbon nitride tubes (HCNT) as photoactive material was prepared for the detection of human epidermal growth factor receptor 2 (HER2). Magnetic Fe 3 O 4 nanospheres (MNs) modified with anti-HER2 antibodies were employed for highly efficient capture of HER2 from serum sample, and Co 3 O 4 nanoparticles (Co 3 O 4 NPs) modified with ascorbic acid oxidase (AAO) as well as HER2 aptamer were used for signal amplification. When the aptamer-Co 3 O 4 -AAO probe was captured onto the electrode surface through the specific binding of the aptamer with HER2, the photocurrent intensity decreased. This was because Co 3 O 4 NPs competed with HCNT for consumption of the excitation energy. As a consequence AAO catalyzed the oxidation of the electron donor (AA), and the aptamer-Co 3 O 4 -AAO probe increased the steric hindrance at the electrode surface, leading to significant photocurrent intensity decrease, thus realizing multiple signal amplification. Based on this signal amplification strategy, at 0 V (vs Ag/AgCl), the PEC sensor shows a wide linear response ranging from 1 pg mL -1 to 1 ng mL -1 with a low detection limit of 0.026 pg mL -1 for HER2. Importantly, the prepared PEC sensor was applied for detection of HER2 in human serum samples with recoveries between 98.8 and 101%. Sensitive photoelectrochemical sensor based on Co 3 O 4 nanoparticles modified with ascorbic acid oxidase for signal amplification is reported.

Three disposable stochastic sensors using nanolayer deposition of a graphene nanocomposite comprising graphene nanoparticles and gold nanoparticles, on different supports: silk, plastic, and paper, and modified with chitosan, were characterized and validated for molecular recognition and quantification of maspin in biological samples. Very low limits of determination (of pg mL -1 magnitude order) were recorded (5.12 pg mL -1 for the sensor based on silk at pH 7.40 and on copy paper at pHs 3.00 and 7.40; 16 ng mL -1 at pH 7.40 for the sensor based on plastic, 41.00 pg mL -1 for the sensor based on silk at pH 3.00, and 204.00 pg mL -1 for the sensor based on plastic, at pH 3.00), with wide linear concentration ranges (5.12 × 10 -12 -2.00 × 10 -6  g mL -1 for the sensors based on silk at pH 7.40, and on copy paper at pH 3.00; 5.12 × 10 -12 -8.00 × 10 -7  g mL -1 for the sensor based on copy paper at pH 7.40; 1.60 × 10 -8 -2.00 × 10 -6  g mL -1 for the sensor based on plastic at pH 7.40; 4.10 × 10 -14 -2.00 × 10 -6  g mL -1 for the sensor based on silk, at pH 3.00; and 2.04 × 10 -13 -8.00 × 10 -7  g mL -1 for the sensor based on plastic at pH 3.00) allowing the molecular recognition and quantification of maspin in healthy people and patients with gastric cancer, when a potential of 125 mV vs. Ag/AgCl was applied. The recoveries of maspin in whole blood, saliva, urine, and tissue samples were higher than 95.00%, with a relative standard deviation lower than 1.0%., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Low oxidation stability is the main drawback of biodiesels and biokerosenes that is overcome by using antioxidants, which can be combined due to synergistic effects. This paper demonstrates that 3D-printed electrochemical devices can be applied to biofuel electroanalysis, including the monitoring of oxidation stability by quantifying the antioxidant content in biofuels. Fabrication requires 3D-printed acrylic templates at which a polylactic acid (PLA) filament with conducting carbon-black filling sensors is extruded by a 3D pen. The antioxidants butyl hydroxyanisole (BHA) and tert-butylhydroquinone (TBHQ) are the most employed additives in biodiesel production, and thus, their electrochemical behavior was investigated; 2,6-ditertbutylphenol (2,6-DTBP) was included in this investigation because it is commonly added to biokerosenes. The electrochemical surface treatment of the 3D-printed electrodes improved the current responses of all antioxidants; however, the electrochemical oxidation of TBHQ was clearly more affected by an electrocatalytic action shifting its oxidation towards less positive potentials (~200 mV), which resulted in a better separation of TBHQ and BHA oxidation peaks (+0.4 and +0.6 V vs Ag|AgCl, respectively). The oxidation of 2,6-DTBP occurred at more positive potentials (+1.2 V vs Ag|AgCl). The simultaneous determination of TBHQ and BHA by differential-pulse voltammetry resulted in linear responses in the range 0.5 and 175 μmol L -1 with limits of detection and quantification of 0.15 μmol L -1 and 0.5 μmol L -1 , respectively. The presence of Fe 3+ , Cu 2+ , Pb 2+ , Mn 2+ , Cd 2+ , and Zn 2+ , even in high concentrations, did not interfere in the determination of TBHQ and BHA. The determination of 2,6-DTBP in biokerosene was achieved by cyclic voltammetry. All relative standard deviations (RSD) were lower than 6.0 %, indicating adequate precision of the methods. Spiked biofuel samples were analyzed (after dilution in electrolyte) and recovery values between 85 and 120% were obtained, which indicates absence of sample matrix effects., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

Aptamers are short single-stranded oligonucleotides (either DNA or RNA) that can fold into well-defined three-dimensional (3D) spatial structures which enable them to capture their specific target by complementary shape interactions. Aptamers are selected from large random libraries through the SELEX process and only a small fraction of the sequence is involved in direct docking with the target. In this paper, we describe the possible truncation variants of zearalenone (ZEA) aptamer which might be an effective binding region for the target. The originally selected zearalenone (ZEA) aptamer was 80-mer in length and shown to bind the target with a high affinity (K d  = 41 ± 5 nM). Herein, computational docking simulation was performed with 15 truncated variants to determine the predicted binding energy and responsible binding site of the aptamer-analyte complex. The results revealed that 5 truncated variants had binding energy lower than - 7.0 kcal/mol. Circular dichroism analysis was performed on the shortlisted aptamer and the conformational change of aptamers was observed with the presence of an analyte. Aptamer Z3IN (29-mer) was chosen as the most enhanced affinity for its target with a dissociation constant of 11.77 ± 1.44 nM. The aptamer was further applied in the electrochemical aptasensor of ZEA based on an indirect competitive format. The results demonstrated that the truncated aptamer leads to an enhancement of the sensitivity of the biosensor.