Simultaneous Detection of Citric Acid and Oxalic Acid Based on Dual Spectrum and Biomimetic Peroxidase for Urolithiasis Screening with a Fully Automatic Urine Analyzer.

Urolithiasis stands as a prevalent ailment within the urinary system, with hyperoxaluria and hypocitraturia being the most frequent manifestations characterized by excessive oxalic acid (OA) and deficient citric acid (CA) levels in urine. Detecting these compounds in urine quantitatively holds paramount importance for early urolithiasis screening. Existing methodologies fall short in achieving simultaneous and on-site identification of OA and CA, posing challenges for accurate urolithiasis screening. Addressing this concern, the study successfully accomplishes the concurrent identification of OA and CA in urine through a combination of dual-spectral analysis and biomimetic peroxidase utilization. Bovine serum albumin and dithiothreitol-modified copper nanoclusters (BSA-DTT-CuNCs) are employed as biomimetic peroxidases, effectively mitigating interference and enabling the simultaneous determination of OA and CA. The quantification range spans from 0 to 12 mm for OA and 0.5 to 2.5 mm for CA, with detection limits of 0.18 and 0.11 mm, respectively. To facilitate swift and on-location urine analysis, a fully automated urine analyzer (FAUA) is introduced that streamlines the process of biomarker pretreatment and identification within urine samples. Validation with real urine samples from urolithiasis patients demonstrates the method's diagnostic precision, highlighting the dual-spectral technique and analyzer's promising role in urolithiasis screening.

A robust colorimetric aptasensor for the label-free detection of marine toxins based on tyrosine-capped gold nanoparticles.

PbTx-2 and okadaic acid (OA) are two typical marine toxins that are highly toxic and harmful to human health. The approach based on citrate-capped gold nanoparticles (Cit-AuNPs) and specific aptamers to construct label-free colorimetric sensors is a widely used method for marine toxin detection. However, the potential interactions between Cit-AuNPs and target molecules have always been ignored, which may result in wrong analytical results due to shortcomings in the Cit-AuNPs. To overcome these shortfalls, in this work, AuNPs were synthesized using tyrosine as a reducing and capping agent, and a robust colorimetric aptasensor based on tyrosine-capped AuNPs (Tyr-AuNPs) was constructed for the label-free detection of marine toxins. Tyr-AuNPs presented better stability compared to Cit-AuNPs due to the stronger binding of amine groups on tyrosine to AuNPs through the Au-N bond. Interactions between Tyr-AuNPs and PbTx-2 were analyzed through UV-vis and isothermal titration calorimetry methods and the results validated the robustness of the Tyr-AuNPs. Colorimetric aptasensors were established for PbTx-2 and OA detection with a linear range of 0.05-4 ppm and limits of detection of 2.25 ppb and 5.19 ppb, respectively. These results demonstrate that the developed colorimetric aptasensor can be a robust and promising method for marine toxin detection.

Label-free and highly-sensitive detection of calcium ions using a silicon-on-sapphire light-addressable potentiometric sensor.

BACKGROUND: Ionic calcium (Ca2+) plays a crucial role in maintaining normal physiological and biochemical functions within the human body. Detecting the concentration of Ca2+ is of utmost significance for various purposes, including disease screening, cellular metabolism research, and evaluating drug effectiveness. However, current detection approaches such as fluorescence and colorimetry face limitations due to complex labeling techniques and the inability to track changes in Ca2+ concentration. In recent years, extensive research has been conducted in this field to explore label-free and efficient approaches. RESULTS: In this study, a novel light-addressed potentiometric sensor (LAPS) using silicon-on-sapphire technology, has been successfully developed for Ca2+ sensing. The Ca2+-sensitive LAPS achieved a wide-range detection of Ca2+, ranging from 10-2 M to 10-7 M, with an impressive detection limit of 100 nM. These advancements are attributed to the ultra-thin silicon layer, silicon dioxide layer, and solid-state silicon rubber sensitive membrane around 6 µm. Furthermore, the sensor demonstrated the ability to dynamically monitor fluctuations in Ca2+ concentration ranging from 10-9 M to 10-2 M within a solution. Its remarkable selectivity, specificity, and long-term stability have facilitated its successful application in the detection of Ca2+ in human serum and urine. SIGNIFICANCE AND NOVELTY: This work presents a Ca2+-sensitive sensor that combines a low detection limit and a wide detection range. The development represents the emergence of a label-free and rapid Ca2+ detection tool with immense prospects in home-based health monitoring, community disease screening, as well as cellular metabolism, and drug screening evaluations.

Engineering a universal and fully automatic analyzer for multichannel and on-site biochemical detection in body fluids.

OBJECTIVE: Rising concerns over wellness and aging have heightened the demand for convenient and efficient on-site health monitoring and disease screening. Current research, focused on specific biomarker detection, often neglects the complexities of sample matrix interference and the absence of a comprehensive, automated platform. To address these issues, we have developed a universal, fully automated analyzer for multifaceted, on-site biochemical analysis of body fluids. METHODS: This analyzer integrates automated sample pretreatment, automatic dilution, detection, and self-cleaning functionalities seamlessly. It is designed to detect a wide range of analytes, from small molecules to macromolecules, including ions and proteins, utilizing spectrophotometric sensing. After optimization, the analyzer achieves performance comparable to traditional Enzyme-Linked Immunosorbent Assay (ELISA), while significantly expanding its detection range through automated dilution. RESULTS: Demonstrations of small molecule detection include the simultaneous assessment of citric acid (CA) and oxalic acid (OA) in urine, achieving recovery rates between 96.65%-106.42% and 93.13%-112.50%, respectively. For protein detection, the analyzer successfully identified Cyfra21-1 in saliva with a recovery rate of 104.93%-111.31%. The pre-treatment process requires only 8.8 minutes, showing enhanced recovery rates for CA and OA at 97.8% and 97.6% respectively, which are superior and more rapid than manual methods. CONCLUSION: The exemplary pretreatment and detection performance of the analyzer underlines its effectiveness in multifaceted, on-site biomarker detection, establishing it as a promising and versatile tool for disease screening and health monitoring. SIGNIFICANCE: This analyzer offers a powerful technological solution for on-site fluid testing, advancing community health care by facilitating more reliable and rapid diagnostics.

A light-addressable potentiometric sensor-based extracellular calcium dynamic monitoring and imaging platform for cellular calcium channel drug evaluation.

Disruption and dysregulation of cellular calcium channel function can lead to diseases such as ischemic stroke, heart failure, and arrhythmias. Corresponding calcium channel drugs typically require preliminary efficacy evaluations using in vitro models such as cells and simulated tissues before clinical testing. However, traditional detection and evaluation methods often encounter challenges in long-term continuous monitoring and lack calcium specificity. In this study, a dynamic monitoring system based on ion-sensitive membranes for light-addressable potentiometric sensor (LAPS) was developed to meet the demand for monitoring changes in extracellular calcium ion (Ca2+) concentration in live cells. The effects of Ca2+ channel agonists and blockers on 2D and 3D HL-1 cells were investigated, with changes in extracellular Ca2+ concentration reflecting cellular calcium metabolism, facilitating drug evaluation. Additionally, calcium imaging technology with optical addressing capability complemented the LAPS system's ability to perceive 3D cell morphology, enhancing its drug evaluation capabilities. This work provides a novel, label-free, specific, and stable technique for monitoring cellular calcium metabolism. It achieves both continuous monitoring at single points and custom sensing area calcium imaging, holding significant implications for drug screening and disease treatment related to human calcium homeostasis.

Enzyme-free and wide-range portable colorimetric sensing system for uric acid and hydrogen peroxide based on copper nanoparticles.

Uric acid (UA) is the final product of purine metabolism. A high concentration of UA in body fluid may lead to kidney stones, gout, and some cardiovascular diseases. Therefore, the non-invasive daily monitoring of UA is of great significance for both hyperuricemia patients and fit people. However, most of the current detection methods for UA are enzyme-dependent which limits the application scenarios and lacks portable instruments for on-site detection, including optics and electrochemistry. In this work, an enzyme-free and wide-range colorimetric sensor for UA and H2O2 detection was developed based on a mercaptosuccinic acid (MSA)-modified Cu nanoparticles (CuNPs). Under the action of UA or H2O2, with the cleavage of MSAs on the CuNPs surface, small Cu particles are further aggregated into larger particles with a lightning violet color. With the employment of the multi-channel handheld automatic photometer (MHAP), the concentration of UA and H2O2 can be determined on-site according to the absorbance measurement by the photodiodes. The linear range of UA was 5 µM-4.5 mM with the limit of detection (LOD) of 3.7 µM, while the linear range of H2O2 was 5 mM-500 mM and 5 µM-5 mM with the LOD of 4.3 µM. This approach has been applied to the detection of UA in human urine, providing more possibilities for non-invasive home health monitoring, community medical diagnosis, and broader prospects of on-site disease detection.

Progress in optical sensors-based uric acid detection.

The escalating number of patients affected by various diseases, such as gout, attributed to abnormal uric acid (UA) concentrations in body fluids, has underscored the need for rapid, efficient, highly sensitive, and stable UA detection methods and sensors. Optical sensors have garnered significant attention due to their simplicity, cost-effectiveness, and resistance to electromagnetic interference. Notably, research efforts have been directed towards UA on-site detection, enabling daily monitoring at home and facilitating rapid disease screening in the community. This review aims to systematically categorize and provide detailed descriptions of the notable achievements and emerging technologies in UA optical sensors over the past five years. The review highlights the advantages of each sensor while also identifying their limitations in on-site applications. Furthermore, recent progress in instrumentation and the application of UA on-site detection in body fluids is discussed, along with the existing challenges and prospects for future development. The review serves as an informative resource, offering technical insights and promising directions for future research in the design and application of on-site optical sensors for UA detection.

Single-stranded DNA binding protein coupled aptasensor with carbon-gold nanoparticle amplification for marine toxins detection assisted by a miniaturized absorbance reader.

Okadaic acid (OA), one of the most widely distributed marine toxins worldwide poses a severe threat to human health. Previous sensing methods for OA detection are usually based on antigen-antibody binding mechanism. However, the drawbacks of antibodies especially the enzyme-labeled antibodies, such as the harsh storage condition and high cost, lead to significant challenges to OA detection in biological samples. To overcome these limitations, a single-stranded DNA binding protein (SSB) coupled aptasensor was developed for OA detection. SSB was incubated on the microplate as a substitute for conventional OA-protein conjugations. Carbon-gold nanoparticles were synthesized and labeled with horseradish peroxidase and thiol-modified aptamers to obtain a capture probe (CGNs@HRP-Apt) instead of the enzyme-labeled antibody for signal amplification. OA and SSB competed to bind with limited aptamers on CGNs@HRP-Apt probes followed by colorimetric assay to obtain the optical signals correlated to OA concentration. To achieve on-site detection, a miniaturized and multichannel absorbance reader (Smart-plate reader) was self-designed with full automation for OA detection. Utilizing the SSB coupled aptasensor and the Smart-plate reader, our approach enables cost-effective and on-site OA sensing with a detection range of 2.5-80 ppb and an ultra-low limit of detection of 0.68 ppb. Moreover, novel OA detection kits based on the SSB coupled aptasensor were prepared which can effectively reduce the cost by 15 times lower than that of commercial ELISA kits. Therefore, the developed platform provides a favorable and promising avenue for marine toxin detection in aquaculture and food safety.

Evaluating the efficacy and cardiotoxicity of EGFR-TKI AC0010 with a novel multifunctional biosensor.

Non-small cell lung cancer (NSCLC) is a leading cause of cancer mortality worldwide. Although epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) have dramatically improved the life expectancy of patients with NSCLC, concerns about TKI-induced cardiotoxicities have increased. AC0010, a novel third-generation TKI, was developed to overcome drug resistance induced by EGFR-T790M mutation. However, the cardiotoxicity of AC0010 remains unclear. To evaluate the efficacy and cardiotoxicity of AC0010, we designed a novel multifunctional biosensor by integrating microelectrodes (MEs) and interdigital electrodes (IDEs) to comprehensively evaluate cell viability, electrophysiological activity, and morphological changes (beating of cardiomyocytes). The multifunctional biosensor can monitor AC0010-induced NSCLC inhibition and cardiotoxicity in a quantitative, label-free, noninvasive, and real-time manner. AC0010 was found to significantly inhibit NCI-H1975 (EGFR-L858R/T790M mutation), while weak inhibition was found for A549 (wild-type EGFR). Negligible inhibition was found in the viabilities of HFF-1 (normal fibroblasts) and cardiomyocytes. With the multifunctional biosensor, we found that 10 µM AC0010 significantly affected the extracellular field potential (EFP) and mechanical beating of cardiomyocytes. The amplitude of EFP continuously decreased after AC0010 treatment, while the interval decreased first and then increased. We analyzed the change in the systole time (ST) and diastole time (DT) within a beating interval and found that the DT and DT/beating interval rate decreased within 1 h after AC0010 treatment. This result probably indicated that the relaxation of cardiomyocytes was insufficient, which may further aggravate the dysfunction. Here, we found that AC0010 significantly inhibited EGFR-mutant NSCLC cells and impaired cardiomyocyte function at low concentrations (10 µM). This is the first study in which the risk of AC0010-induced cardiotoxicity was evaluated. In addition, novel multifunctional biosensors can comprehensively evaluate the antitumor efficacy and cardiotoxicity of drugs and candidate compounds.

Multifunctional AuNPs@HRP@FeMOF immune scaffold with a fully automated saliva analyzer for oral cancer screening.

Delayed diagnosis of cancer-causing death is a worldwide concern. General diagnosis methods are invasive, time-consuming, and operation complicated, which are not suitable for preliminary screening. To address these challenges, the sensing platform based on immune scaffold and fully automated saliva analyzer (FASA) was proposed for oral cancer screening for the first time by non-invasive detection of Cyfra21-1 in saliva. Through one-step synthesis method with unique covalent and electrostatic adsorption strategy, AuNPs@HRP@FeMOF immune scaffold features multiple functions including antibody carrier, catalytic activity, and signal amplification. Highly integrated FASA with the immune scaffold provides automatic testing to avoid false-positive results and reduce pretreatment time without any user intervention. Compared with the commercial analyzer, FASA has comparable performance for Cyfra21-1 detection with a detection range of 3.1-50.0 ng/mL and R2 of 0.971, and superior features in full automation, high integration, time saving and low cost. Oral cancer patients could be distinguished accurately by the platform with an excellent correlation (R2 of 0.904) and average RSD (5.578%) without sample dilution. The proposed platform provides an effective and promising tool for cancer screening in point-of-care applications, which can be further extended for biomarker detection in universal body fluids, disease screening, prognosis review and homecare monitoring.

Colorimetric detection of citric acid as the biomarker for urolithiasis based on sodium dodecylsulfate-AgNPs with a portable CD-spectrometer.

Citric acid (CA) has been considered as a biomarker of urolithiasis due to its vital suppression role in urinary stone formation. Most analytical methods for detecting CA are complicated and require expensive equipment. Herein, a colorimetric method based on sodium dodecylsulfate (SDS) modified AgNPs was presented for convenient and portable detection of CA in urine. By detecting the absorption of the solution color, the quantitative detection of CA can be achieved. The pH value of SDS-AgNPs, concentration of Al3+ and incubation time were optimized. Under optimal conditions, the method has the detection range of 1-10 mg/L, with a limit of detection (LOD) of 0.21 mg/L. Moreover, a self-developed portable CD(Compact Disk)-spectrometer (CDs) was established for detecting CA with a LOD of 0.49 mg/L featuring high simplicity, low time cost and good portability. This method was also validated with good selectivity to CA. In addition, the artificial urine samples were also detected to verify the capability of the method and CDs. The results validated that the method integrating with the CDs can be a promising platform for citric acid detection that can be further used for early screening and prognostic monitoring of urolithiasis.

Development of QDs-based nanosensors for heavy metal detection: A review on transducer principles and in-situ detection.

Heavy metal pollution has severe threats to the ecological environment and human health. Thus, it is urgent to achieve the rapid, selective, sensitive and portable detection of heavy metal ions. To overcome the defects of traditional methods such as time-consuming, low sensitivity, high cost and complicated operation, QDs (Quantum dots)-based nanomaterials have been used in sensors to significantly improve the sensing performance. Due to their excellent physicochemical properties, high specific surface area, high adsorption and reactive capacity, nanomaterials could act as potential probes or offer enhanced sensitivity and create a promising nanosensors platform. In this review, the rapidly advancing types of QDs for heavy metal ions detection are first summarized. Modified with ligands, nanomaterials, or biomaterials, QDs are assembled on sensors by the interaction of electrostatic adsorption, chemical bonding, steric hindrance, and base-pairing. The stability of QDs-based nanosensors is improved by doping the elements to QDs, providing the reference substance, optimizing the assemble strategies and so on. Then, according to transducer principles, the two most typical sensor categories based on QDs: optical and electrochemical sensors are highlighted to be discussed. In the meanwhile, portable devices combining with QDs to adapt the practical detection in complex situations are summarized. The deficiencies and future challenges of QDs in toxicity, specificity, portability, multi-metal co-detection and degradation during the detection are also pointed out. In the end, the development trends of QDs-based nanosensors for heavy metal ions detection are discussed. This review presents an overall understanding, recent advances, current challenges and future outlook of QDs-based nanosensors for heavy metal detection.

A multi-channel handheld automatic spectrometer for wide range and on-site detection of okadaic acid based on specific aptamer binding.

Okadaic acid (OA) is one of the marine toxins that are widely distributed and harmful to humans. However, the current detection methods for OA involve complex procedures, need long detection time, and rely on large-scale laboratory equipment. In this work, a multi-channel handheld automatic spectrometer (MHAS) based on a spectral sensor was developed with the advantages of small size, simple operation and low cost. It could achieve rapid detection within 30 s and a wide spectral detection range of 470-780 nm with a broadband LED as the light source and a microplate containing 8 wells as a sample cell. Moreover, through the combination of gold nanoparticles (AuNPs) and aptamer-OA34, a highly sensitive and rapid system for OA detection was established with a LOD of 1.80 µg L-1 and a wide detection range of 20-10 000 µg L-1, which is comparable to a microplate reader. Compared with other studies, the proposed MHAS realized rapid on-site detection of OA with a wider detection range, shorter detection time and higher portability. Therefore, the MHAS promises to be a stable and efficient optical detection instrument for on-site detection in the fields of food safety, disease diagnosis and environmental monitoring.

A novel smartphone-based CD-spectrometer for high sensitive and cost-effective colorimetric detection of ascorbic acid.

As a powerful tool for medical diagnosis and bioanalysis, conventional optical spectrometers are generally expensive, bulky and always require an accompanying data processing device. In this work, we developed a novel smartphone-based CD-spectrometer (SCDS) for high sensitive and ultra-portable colorimetric analysis, with the advantage of cost-effective and simplicity. The distance between the light source and slit, the structure of SCDS and the parameters of camera in the smartphone were all optimized to ensure the best analytical performance. Besides, the SCDS employed HSV color model and utilized the overall intensity calculated by summing V-value of adjacent position for the absorbance measurement. In this way the errors caused by the low resolution of CD-grating can effectively be eliminated to promote the sensitivity of the SCDS. The performance of the SCDS was first validated for colorimetric detection of BSA with a detection limit of 0.0073 mg/mL, which is superior compared to that of the microtiter plate reader (MTPR). Moreover, by combining with 3,3',5,5'-tetramethylbenzidine-manganese dioxide (TMB-MnO2) nanosheets reaction, a high sensitive and specific system for ascorbic acid detection was established. The SCDS gives a detection range from 0.6250 µM to 40 µM with a detection limit of 0.4946 µM for AA detection. Compared to other studies, the SCDS features wide detection range and very low detection limit with low cost instrument. Therefore, the SCDS will be an ideal and promising colorimetric system for point-of-care (POC) application in food security, disease diagnosis and environmental monitoring.

Microfluidic-based fluorescent electronic eye with CdTe/CdS core-shell quantum dots for trace detection of cadmium ions.

Cadmium ions (Cd2+) greatly threat human health and the environment due to its extremely severe toxicity. Therefore, it is of paramount importance to establish a sensitive and portable platform for monitoring Cd2+ on site. In this work, a novel microfluidic-based fluorescent electronic eye (E-eye) combined with tetrasodium iminodisuccinate (IDS)-etched CdTe/CdS quantum dots (QDs) was developed for portable and sensitive detection of trace Cd2+ in water environment. The fluorescent E-eye consists of a microfluidic chip for miniaturized flow analysis, an ultraviolet light excitation module for fluorescent excitation, an optical lens for device miniaturization and a smartphone for portable photographing and analysis. The IDS was added in the CdTe/CdS QDs to cause fluorescence quenching due to the chemical etching. Subsequently added Cd2+ will be recognized by etched QDs, thus inducing the fluorescence changes that can be directly captured by the E-eye for quantitative detection of Cd2+. With the optimization of all parameters including pH, reaction time and the concentration of IDS, the proposed platform could detect Cd2+ with a low detection limit of 0.26 µg/L in the range of 1-250 µg/L. It is worth noting that the performance of the developed fluorescent E-eye is quite comparable to a commercial microplate reader with a detailed comparison in linearity, sensitivity and detection limit. In summary, the proposed microfluidic-based fluorescent E-eye provides a promising platform for portable and high sensitive detection of trace cadmium in water environment.

A novel portable biosensor based on aptamer functionalized gold nanoparticles for adenosine detection.

Adenosine has received great attentions acting as a potential biomarker for monitoring lung cancer. Most of the reported studies for adenosine detection require large instruments and complicated procedures. Herein, a sensitive, rapid and in-situ colorimetric aptasensor was developed for adenosine detection. Moreover, a homemade biomimetic electronic-eye (E-eye) was established and utilized as a portable in-time detection equipment. The entire measurement can be completed within 20 min, including the combination of aptamer with adenosine or AuNPs and the detection of adenosine. Four different kinds of aptamer were compared and the results showed that the AuNPs-aptamer-biotin system was the most stable and with the widest detection range of 5.0 µM-60.0 µM and the lowest LOD of 0.17 µM. Moreover, the artificial urine samples were also tested with a linear range from 5.0 to 50.0 µM and a LOD of 0.48 µM. The results validated that the aptasensor together with the E-eye can be a promising platform for adenosine detection.