The detection of epidermal growth factor receptor (EGFR) mutation L858R in circulating tumor DNA (ctDNA) is beneficial for the clinical diagnosis and personalized therapy of non-small cell lung cancer (NSCLC). Herein, for the first time, the combination of the primer exchange reaction (PER) and clustered regularly interspaced short palindromic repeats (CRISPR) and its associated nucleases (Cas) 14a was used in electrochemical biosensor construction for the detection of ctDNA EGFR L858R. EGFR L858R, as the target, induced the isothermal amplification of the PER reaction, and then the CRISPR/Cas14a system was activated; subsequently, the substrate ssDNA-MB was cleaved and the electron on the surface of the gold electrode transferred, resulting in the fluctuation of the electrochemical redox signal on the electrode surface, whereas the electrochemical signal will be stable when EGFR L858R is absent. Therefore, the concentration of EGFR L858R can be quantified by electrochemical signal analysis. The low detection limit is 0.34 fM and the dynamic detection range is from 1 fM to 1 μM in this work. The PER-CRISPR/Cas14a electrochemical biosensor greatly improved the analytical sensitivity. In addition, this platform also exhibited excellent specificity, reproducibility, stability and good recovery. This study provides an efficient and novel strategy for the detection of ctDNA EGFR L858R, which has great potential for application in the diagnosis and treatment of NSCLC.

A DNOC electrochemical sensor has been developed by using a composite of Zr-UiO-66 and FMWCNTs on a glassy carbon electrode (GCE) and using the differential pulse voltammetry technique. The synthesized materials were physico-chemically characterized by BET, PXRD, FTIR, TGA, EDX, and FESEM. Cyclic voltammetry showed that DNOC has three oxidation peaks at 0.03 V (RSD: 0.23%), 0.42 V (RSD: 0.21%), and 1.32 V (RSD: 0.32%) and three reduction peaks at – 0.20 V (RSD: 0.15%), – 0.82 V (RSD: 0.26%), and – 1.14 V (RSD: 0.19%) which follow a diffusion-controlled mechanism. Different parameters were optimized using differential pulse voltammetry and good linear ranges were found for the simultaneous detection of the three reduction peaks. For a specific concentration range of 0.1–50 μM, a limit of detection of 0.119 μM based on 3Sb/m was obtained. The interfering effects of five non-phenolic pesticides and five heavy metals were evaluated to highlight the selectivity of the developed sensor. It is the first report of an electrochemical DNOC sensor in which all three oxidation peaks are prominently visible. Ethion and chloropyriphos were found to inhibit the redox process of DNOC on the developed sensor platform Zr-UiO-66/FMWCNT/GCE. The sensor was successfully applied to DNOC determination in spiked potato samples and the results showed a standard deviation of less than 3%. The proposed method is expected to provide a novel platform for the quantitative determination of DNOC pesticides in vegetables.

A study on the inactivation and germination mechanism of spores is very important in the application of spores, as such high-purity spores are the basis of related research. However, spores and vegetative cells of bacteria often coexist, and it is difficult to separate them. In this study, a magnetic flow device for the purification of spores in the culture medium system was developed based on a “stepped” structure with a magnetic force that could absorb vegetative cells with magnetic nanoparticles. The operation process was as follows: first, vancomycin functionalized nanoparticles were used to prepare Van-Fe3O4 NPs, which were then combined with vegetative cells to form a magnetic conjugate. Subsequently, the magnetic conjugate (vegetative cells) flowed through the “stepped” magnetic flow device and was adsorbed. Meanwhile, the spores moved through the channel and were collected. The achieved purity of the collected spores was more than 95%. Further, the number of the obtained spores was quickly quantified using Raman spectroscopy. The entire purification and quantitative process can be completed within 30 min and the limit of detection was 5 CFU mL−1. This study showed outstanding spore purification ability and provided a new method for purification and rapid quantitative detection of spores.

A metal–organic framework (MOF) is a good carrier for molecular imprinting due to its high surface area and strong adsorption capacity, but its poor dispersibility in aqueous solution is one of the significant drawbacks, which can severely impede its effectiveness. Amphiphilic block copolymers are good hydrophilic materials and have the potential to overcome the shortcomings of MOFs. In order to improve the hydrophilicity of molecularly imprinted fluorescent materials, we have applied a combination of molecularly imprinted technology and amphiphilic block copolymers on MOFs for the first time. Amphiphilic PAVE copolymer is selected as the molecular imprinted functional monomer to improve the hydrophilicity of UiO-66-NH2. The synthesized PAVE-MOF-MIP has adequate water dispersion ability and fluorescence activity. When encountering oxytetracycline, PAVE-MOF-MIP will produce fluorescence quenching, it is used to construct a fluorescence detection platform for oxytetracycline detection. Compared with traditional MIP@MOF, PAVE-MOF-MIP has better water dispersion ability and detection accuracy. Under optimal conditions, the linear range of oxytetracycline detection is 10–100 μmol L−1, and the minimum limit of detection (LOD) is 86 nmol L−1. This paper proposes a novel approach to use amphiphilic block copolymers as molecularly imprinted monomers on MOFs, providing an innovative idea that has not been previously explored.

The first page of this article is displayed as the abstract.

The first page of this article is displayed as the abstract.

In this study, we developed an ultra-performance liquid chromatography-quadrupole-Orbitrap mass spectrometry (UPLC-Q-Orbitrap-MS) method for the analysis of seven steroid hormones in human tears. Tear samples were collected using Schirmer strips and extracted with methanol. The analytes were then subjected to a “one-step” clean-up process using solid phase extraction, and subsequently separated on a C18 column by UPLC. Detection was performed using an Orbitrap MS detector, operated at a resolution of 17 500 FWHM in parallel reaction monitoring mode with an HESI ion source under positive ionization. Our data showed the sensitivity with limits of detection for steroid hormones in tears ranging from 0.12 to 0.86 pg μL−1, and high correlation coefficients in the corresponding concentration range exceeding 0.99. The results also had high accuracy with spiking recoveries for spiked tear samples ranging from 78.2% to 96.7% and relative deviations of less than 15%. Furthermore, we successfully applied our method to detect the pg μL−1 level of steroid hormones in real human tear samples. Our findings showed the potential of this UPLC-Q-Orbitrap-MS method for the accurate and sensitive determination of steroid hormones in human tears, providing a valuable tool for ophthalmic and endocrine research.

The reliable and accurate detection of unauthorized additives in food is significant to prevent health risks. In recent years, surface-enhanced Raman spectroscopy (SERS) with fast, simple, and sensitive capabilities has been widely used for food safety analysis. In order to detect illegally added dye molecule rhodamine 6G and small molecule melamine in food, we have proposed a fast, convenient, and label-free SERS detection technology using flower-like Ag/ZnO as the SERS substrate. The structure and morphology of the flower-like Ag/ZnO were characterized by scanning electron microscopy, energy dispersive spectrometery, transmission electron microscopy, and X-ray diffraction analysis. We investigated the SERS effect and sensitivity of flower-like Ag/ZnO toward rhodamine 6G and melamine. The synergistic effect of flower-like Ag/ZnO provides high SERS activity for the detection of rhodamine 6G and melamine at the lowest detection concentrations of 0.5 ng mL−1 and 1.0 ng mL−1, respectively. Therefore, flower-like Ag/ZnO with good sensitivity and uniformity has potential for improving the detection of illegal food additives.

In this study, a poly(N-methyl aniline)-cerium oxide-functionalized MWCNTs (PNMA-CeO2-fMWCNTs) composite was synthesized in a one-step preparation technique. As a highly efficient modifier, the composite was used to modify the glassy carbon electrode surface for simultaneous detection of uric acid (UA) and 5-fluorouracil (5-FU). Morphological characterization of the GCE/PNMA-CeO2-fMWCNTs was studied using scanning electron microscopy. Structural characterization of the composite was performed using X-ray diffraction and Fourier-transformed infrared spectroscopy. Electron transfer properties of the prepared electrodes were carried out with electrochemical impedance spectroscopy and cyclic voltammetry. The linear working range for UA and 5-FU was found to be 0.25–50 μM and 0.5–750 μM, respectively. The limit of detection values for UA and 5-FU were 0.04 μM and 0.19 μM, respectively. The effects of various interfering substances on the electrochemical response of UA and 5-FU were investigated. The GCE/PNMA-CeO2-fMWCNTs sensor has excellent stability, reproducibility, anti-interference ability, and reproducibility. To demonstrate the practical application of the sensing platform, fetal bovine serum was selected and tested in the spiked samples, and satisfactory results were obtained. The prepared composite proved to be a promising platform for simple, rapid, and simultaneous analysis of UA and 5-FU.

Based on the original position statistic distribution analysis technique, the characterization method of segregation for large-size metal materials gives significant guidance to the research of material properties and production. However, random errors are inevitably brought into the calculation of segregation degree for materials characterization by the Spark Mapping Analysis for Large Sample (SMALS) technique, resulting in a misguide of the segregation degree. In this paper, we present the lower limit of segregation degree (Ds(L)) method to distinguish the random error from metal material segregation for large-size samples over the SMALS method. The random error of standard material in the 95% confidence interval was utilized as Ds(L) and the method has been applied for macro-segregation quantitative analysis. The precision correlation between Spark Atomic Emission Spectrometry (Spark-AES) and SMALS was established. Furthermore, the functional relationship between the Ds(L) and element content C can be obtained in the SMALS method. The Ds(L) method as the criterion can be used to not only characterize the minimum limit of the segregation degree but also the segregation existence for large-size samples. Applying to low-alloy steel can illustrate the effective performance of the Ds(L) method. Results on both spark mapping analysis and Spark-AES verify the substantial consistency.

Methyltestosterone is one of the banned drugs in aquaculture, and it should be monitored in food-producing animals. 17α-Methyl-5β-androstane-3α,17β-diol, as the main metabolite of methyltestosterone in vertebrates, could be used as another marker for controlling the administration of methyltestosterone, due to its high residual concentration and slow elimination rate. In this study, an analytical method based on gas chromatography-mass spectrometry (GC-MS) was developed and validated for the simultaneous determination of methyltestosterone and its main metabolite in fish. After pretreatment by liquid–liquid extraction with n-hexane and solid phase extraction with C18 and NH2 columns, the target analytes in the muscle tissues were extracted and concentrated, and the influence of the sample matrix was eliminated. Then, the prepared samples were separated and detected with GC-MS in the selected ion monitoring (SIM) mode. Methyltestosterone-D3 was chosen as the internal standard for quantitation. After optimization, the limits of detection for methyltestosterone and 17α-methyl-5β-androstane-3α,17β-diol were 20 μg kg−1 and 15 μg kg−1, respectively. The limits of quantitation were both 50 μg kg−1. The calibration curves showed good linearity in the concentration range from 50.0 ng mL−1 to 500.0 ng mL−1. The correlation coefficients of methyltestosterone and 17α-methyl-5β-androstane-3α,17β-diol were more than 0.9990. The recoveries of the analytes in real samples were in the range of 99.7–116.6% with the relative standard deviation of 5.2–8.3%. The established method could meet the demand for simultaneous detection of methyltestosterone and its major metabolite, and it could be used to provide more information on the abuse of methyltestosterone in food-producing animals.

Gastric cancer is a prevalent malignant tumor of the gastrointestinal tract accompanied by a high mortality rate; therefore, early gastric cancer screening is critical for improving patient survival. In this study, we present a facile fluorescence immunoassay for highly sensitive screening of pepsinogen I (PG I) based on a one-pot biomimetic mineralization process for the synthesis of gold nanocluster-anchored manganese dioxide (AuNCs/MnO2) nanosheets. MnO2 first quenches the fluorescence of AuNCs through the Förster resonance energy transfer effect, whereas the introduction of ascorbic acid (AA) leads to the decomposition of MnO2 and rapidly recovers the fluorescence of AuNCs. Based on the above principles and phenomena, we developed a sensitive fluorescence immunoassay for the in situ generation of AA to detect PG I. Specifically, in the presence of PG I, the sandwich-type immunoreactivity-enriched alkaline phosphatase-labeled secondary antibody catalyzes the production of AA from the substrate, which enhances the fluorescence intensity. Under optimized conditions, the fluorescence intensity increased linearly with the concentration of PG I (0.05 to 200 ng mL−1) with a limit of detection (LOD) of 0.013 ng mL−1 (S/N = 3). The designed sensing platform has good stability (more than one year) and excellent anti-interference capability and demonstrates satisfactory accuracy for detection in real samples compared to commercial ELISA kits.

A novel multi-functional microfluidic paper-based analytical device (μPAD) integrated with ion imprinted polymers (IIPs) was proposed for specific, portable and low-cost detection of cadmium (Cd(II)) in water. The IIP was grafted on paper and integrated into the μPAD for separation of Cd(II) through multi-layer design. The paper-based screen printed carbon electrode (pSPCE) modified with reduced graphene oxide was fabricated and combined with the μPAD for electrochemical sensing of the separated Cd(II). Reduced graphene oxide (rGO) was prepared via electroreduction on the working electrode surface of the pSPCE (rGO/pSPCE), which provided a sensitization effect with an improved signal for Cd(II) detection. The μPAD developed with the integrated IIP and combined with rGO/pSPCE is able to detect Cd(II) with a linear range from 1 ng ml−1 to 100 ng ml−1 and a detection limit of 0.05 ng ml−1. The accuracy of this μPAD was evaluated with spiked real water samples and compared with that of the inductively coupled plasma mass spectrometry (ICP-MS) method, from which the recovery values ranged from 96.5% to 114.2% with RSDs <10% between the two methods. This μPAD demonstrated its advantages of low-cost, portability, and suitability for highly sensitive detection of Cd(II), making it a valuable tool for on-site analysis.

It is of great significance for the clinical diagnosis of Alzheimer's disease (AD) to achieve the on-site activity evaluation of acetylcholinesterase (AChE), the hydrolase of acetylcholine (ACh). Herein, we have developed a biosensing method endowed with considerable superiority based on the organic–inorganic hybrid composite Eu(DPA)3@Lap with excellent stability and fluorescent properties for this purpose by loading Eu3+ ions and 2,6-dipicolinic acid (DPA) into LAPONITE® (Lap). Through the comprehensive consideration of the specific hydrolysis of acetylthiocholine (ATCh) into thiocholine (TCh) by AChE, the high binding affinity of TCh to copper ion (Cu2+), and the selective fluorescence quenching ability of Cu2+, a simple Eu(DPA)3@Lap-based assay was developed to realize the rapid and convenient evaluation of AChE activity. Owning to the facile signal on-off-on response mode with a clear PET-based sensing mechanism, our assay presents favorable selectivity and sensitivity (LOD of 0.5 mU mL−1). Furthermore, the fluorescent assay was successfully applied for assessing AChE activity in human serum samples and screening potential AChE inhibitors, showing potential for application in the early diagnosis and drug screening of AD, as a new development path of AD therapy.

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The emerging sample pretreatment technique of magnetic solid-phase extraction (MSPE) has drawn the attention of researchers owing to its advantages of less reagent consumption, fast separation/enrichment process, high adsorption capacity, and simple operation. This paper presents a review of synthesis techniques, classification, and analysis procedures for MSPE in the detection of heavy metals in food. Magnetic adsorbents derived from silica, metal oxides, carbon, polymers, etc., are applied for the detection of heavy metals in food. Then, the recent development of the technology of MSPE for the analysis of heavy metal extraction in food is summarized in detail. Finally, the future outlook for the improvement of MSPE is also discussed.

The pretreatment of samples was vital for enhancing the sensitivity and accuracy of analytical methods. An efficient and sensitive method, based on modified QuEChERS with high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) for the simultaneous determination of the 13 sulfonamides (SAs) in soil, was developed. After extraction by sonication with methanol, the clean-up procedure was achieved using QuEChERS with a primary secondary amine (PSA). The quantification of the 13 SAs was performed by HPLC-MS/MS in electrospray ionization (ESI) and multiple reaction monitoring (MRM) modes. Under optimized conditions, the standard solution exhibited good linearity within the range of 0.01–0.5 μg mL−1. The limits of detection and the limits of quantification of the developed method were 0.007–0.030 μg kg−1 and 0.022–0.101 μg kg−1, respectively. The spiked recoveries for the 13 SAs were in the range of 74.5–111.7% with RSD less than 15%. Furthermore, the developed method was successfully applied for the determination of SAs in real soil samples. The above results showed that the proposed method would be an ideal analytical method for SAs in environmental ecological research.

This study focused on creating a SERS composite particle specifically designed for detecting malachite green. We synthesized silver nano-dendritic structures on p-type porous silicon using an external electric field, separating them from the silicon wafer. Ultrasonic crushing yielded irregular silver nanodendrite-modified porous silicon composite particles. Upon being tested in an aqueous solution of malachite green, these composite particles demonstrated significant surface-enhanced Raman scattering effects. Our findings highlight the exceptional performance of the SERS substrate composed of porous silicon and irregular silver nano-dendritic particles. It exhibited high sensitivity, specificity, consistent signal strength, and reliability in detecting trace amounts of malachite green in water. Under ideal conditions, the substrate could detect malachite green at concentrations as low as 10−8 M. Moreover, its swift response to trace amounts of malachite green in fish underscores its potential as an effective Raman detector.

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Brazilian spotted fever (BSF) is a serious disease of medical importance due to its rapid evolution and high lethality. The effectiveness of the treatment mainly depends on the rapid diagnosis, which is currently performed by indirect immunofluorescence and PCR tests, which require high costs and laboratory structure. In order to propose an alternative methodology, we sought to develop an impedimetric immunosensor (IM) based on the immobilization of specific IgY antibodies for IgG anti Rickettsia rickettsii, using blood plasma from capybara (Hydrochoerus hydrochaeris), for characterization, validation and applications of the ready IM. IM selectivity was observed when comparing capybara reagent IgG (IgGcr) readings with non-reagent IgG (IgGnr). A reagent IgG calibration curve was obtained, from which the limits of detection (LOD) and quantification (LOQ) of 1.3 ng mL−1 and 4.4 ng mL−1 were calculated, respectively. The accuracy tests showed that different concentrations of IgGcr showed a maximum deviation of 20.0%, with CI between 90.00% and 95.00%. Intermediate precision tests showed a relative standard deviation of 2.09% for researcher 1 and 2.61% for researcher 2, and the F test showed no significant difference between the recovery values found between the two analysts, since Fcal 1.56 < 5.05 and P-value 0.48 > 0, 05. Therefore, an impedimetric immunosensor was developed to detect anti BSF IgG in capybara blood plasma, which greatly contributes to the improvement of diagnostic tests, cost reduction and ease of execution.