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A paper-based analytical device (PAD) with an integrated composite electrode has been designed and fabricated for non-enzymatic creatinine sensing. Reduced graphene oxide (rGO) was employed to modify the PAD so that it could function as a solid-contact transducer. A new macrocyclic pyrido-hexapeptide derivative was made and used as a special ionophore in the creatinine membrane sensor. The synthesized PAD showed a detection limit of 1.0 µM (S/N = 3) and a potentiometric response towards creatinine throughout a log-linear range of 2.0 µM–10 mM (R2 = 0.9998). The sensor shows significant selectivity for a few related substances, including ephedrine, codeine, ketamine, caffeine, urea, urate, carbinoxamine, and dextromethorphan. It has been established that the testing method is appropriate for the direct potentiometric detection of creatinine in a variety of human urine sample types. When an indicating electrode and a reference electrode are put on the same flexible disposable, this lets applications with a small sample volume be done. For point-of-care creatinine measurement, the developed paper-based analytical equipment is a good choice because it is affordable, easily accessible, and self-pumping (especially when combined with potentiometric detection). [ABSTRACT FROM AUTHOR]

Perchlorate, a hazardous pollutant, is mainly found in untreated wastewater from urban and industrial sites and unregulated surface and groundwater sources. Effective monitoring of perchlorate in water is essential to mitigate its potential harmful effects. Microfluidic systems are evolving as promising technologies for detecting chemical contaminants in water due to their ability to enable rapid analysis with minimal consumption of reagents and samples. The integration of paper-based microfluidic devices with digital imaging has garnered enormous attention from the perspective of developing portable analytical techniques. Nevertheless, there is a need for further exploration to fully realize the potential of these systems. This study aimed to develop and evaluate the performance of a microfluidic paper-based device for measuring perchlorate levels in water samples. Smartphone-based digital imaging was integrated with microfluidic paper-based analytical device to establish a reliable colorimetric method for detecting perchlorate contamination. The results demonstrated successful quantitative estimation of perchlorate levels in water samples using a colorimetric assay based on the methylene blue-perchlorate reaction. Real-time, on-site colorimetric data were collected using a digital smartphone, and image processing methods were used to detect the occurrence of perchlorate in water samples from digital images. The developed approach yielded a broad linear response ranging from 4 to 12 µg/L (R2 = 0.97) for perchlorate detection, with a limit of detection of 3.41 µg/L and a limit of quantification of 10.34 µg/L. The findings underscore the effectiveness of colorimetric analysis and digital imaging for paper-based analytical devices. The limitations of this method include the capability to detect only a single analyte and the requirement for additional steps in image processing to obtain analytical results. Future developments should focus on designing devices for simultaneous detection of multiple contaminants and exploring automated methods of image analysis. [ABSTRACT FROM AUTHOR]

This study investigated the colorimetric response of standard glucose, serum glucose, and nucleic acid assays on various paper surfaces with different wettability, including hydrophilic, hydrophobic, and nearly superhydrophobic surfaces. Water contact angles (WCA) formed by water droplets on each surface were measured using ImageJ software. The hydrophilic surface showed no contact angle, while the hydrophobic and nearly superhydrophobic surfaces exhibited contact angles of 115.667° and 133.933°, respectively. The colorimetric sensitivity of the standard glucose assay was analyzed on these surfaces, revealing enhanced sensitivity on the nearly superhydrophobic surface due to the high molecular crowding effect owing to its non-wetting behavior and eventually confined reaction product at the sample loading zone. The hydrophobic nature of the surface restricts the spreading and diffusion of the reaction product, leading to a controlled and localized concentration of the assay product leading to moderate colorimetric intensity. On the other hand, the hydrophilic surface showed the least enhancement in colorimetric sensitivity; this is attributed to the high wettability of the hydrophilic surface causing the reaction product to spread extensively, resulting in a larger area of dispersion and consequently a lower colorimetric intensity. The measured limit of detection (LOD) for nucleic acid on nearly superhydrophobic surfaces was found to be 16.15 ng/µL, which was almost four-fold lower than on hydrophilic surfaces (60.08 ng/µL). Additionally, the LODs of standard glucose and clinical serum samples were two-fold lower on nearly superhydrophobic surfaces compared to hydrophilic surfaces. Our findings clearly highlight the promising potential of utilizing superhydrophobic surfaces to significantly enhance colorimetric sensitivity in paper-based diagnostic applications. This innovative approach holds promise for advancing point-of-care diagnostics and improving disease detection in resource-limited settings. [ABSTRACT FROM AUTHOR]

The development of simple and accurate detection of uracil-DNA glycosylase (UDG) is of great significance for early clinical diagnosis and biomedical research. Here, we on the first effort introduced the uracil bases into the rolling circle amplification (RCA) reaction to produce the functionalized pure DNA hydrogel (PDH) for UDG detection. During RCA process, methylene blue (MB) molecules as the indicators were encapsulated into PDH. The addition of UDG can remove the uracil bases of PDH to generate abasic sites, which are further cleaved with the assistance of apurinic/apyrimidinic endonuclease (APE), thus resulting in the dissociation of PDH to release blue MB. By combining with the paper analytical devices as the signal readout platform, a colorimetric and electrochemical dual-signal biosensor was constructed for convenient and accurate detection of UDG. The proposed MB@PDH-based dual-signal sensing system exhibited good selectivity and high sensitivity with a detection limit of 6.4 × 10−4 U/mL (electrochemical method). It was also demonstrated that this sensing system showed excellent performance in UDG inhibitor screening, thus providing great potential in UDG-related disease diagnosis and drug discovery.

Recently, with the development of microfluidic chips, attempts to integrate cell culture and biomedical material testing functions into a single chip have increased to supplement experimental animal models. Among the evaluations of biomaterials, immunogenicity testing, a current priority, is attracting attention. In this study, we developed a simple and easy-to-handle immunogenicity-testing cell chip to evaluate the immunogenicity of biomaterials. On this chip, macrophages were introduced as immunogenicity indicators, and a micro-paper-based analytical device (µPAD) was used for optical analysis. Macrophages are present in all parts of the body and mediate immune reactions against body implants or bio-derived substances, resulting in the production of hydrogen peroxide. In the cell chamber of the developed cell chip, macrophages grow and react to immunogenic materials. Activated macrophages secrete hydrogen peroxide, which is then transferred to the PAD with single-finger actuation. The hydrogen peroxide molecules reaching the PAD detection zone react with the colorimetric detection substrate, resulting in a color that corresponds to the hydrogen peroxide concentration. With the developed testing chip, the immunogenicity of biomaterials can be determined before administration. [ABSTRACT FROM AUTHOR]

The separation of enantiomers plays a critical role in pharmaceutical development, ensuring therapeutic efficacy, safety, and patent protection. It enables the production of enantiopure drugs and enhances our understanding of the properties of chiral compounds. In this study, a straightforward and effective chiral detection strategy was developed for distinguishing between tryptophan (TRP) enantiomers. The approach involved the preparation of a magnetic molecularly imprinted chitosan (MMIC) for preparation of the sample, which was combined with a nitrocellulose membrane (a paper-based analytical device, PAD) integrated with D-TRP covalently grafted with polymethacrylic acid (PAD-PMA_D-TRP). Discriminating between the TRP enantiomers was achieved using AuNPs as a colorimetric probe. Indeed, the presence of D-TRP rapidly induced the aggregation of AuNPs due to its strong affinity to PAD-PMA_D-TRP, resulting in a noticeable change in the color of the AuNPs from red to purple. On the other hand, L-TRP did not induce any color changes. The chiral analysis could be easily performed with the naked eye and/or a smartphone. The developed method exhibited a detection limit of 3.3 µM, and it was successfully applied to detect TRP in serum samples, demonstrating good recovery rates. The proposed procedure is characterized by its simplicity, cost-effectiveness, rapidity, and ease of operation. [ABSTRACT FROM AUTHOR]

Loop-mediated isothermal amplification (LAMP) has been widely used to detect many infectious diseases. However, minor inconveniences during the steps of adding reaction ingredients and lack of simple color results hinder point-of-care detection. We therefore invented a fluorometric paper-based LAMP by incorporating LAMP reagents, including a biotinylated primer, onto a cellulose membrane paper, with a simple DNA fluorescent dye incubation that demonstrated rapid and accurate results parallel to quantitative polymerase chain reaction (qPCR) methods. This technology allows for instant paper strip detection of methicillin-resistant Staphylococcus aureus (MRSA) in the laboratory and clinical samples. MRSA represents a major public health problem as it can cause infections in different parts of the human body and yet is resistant to commonly used antibiotics. In this study, we optimized LAMP reaction ingredients and incubation conditions following a central composite design (CCD) that yielded the shortest reaction time with high sensitivity. These CCD components and conditions were used to construct the paper-based LAMP reaction by immobilizing the biotinylated primer and the rest of the LAMP reagents to produce the ready-to-use MRSA diagnostic device. Our paper-based LAMP device could detect as low as 10 ag (equivalent to 1 copy) of the MRSA gene mecA within 36-43 min, was evaluated using both laboratory (individual cultures of MRSA and non-MRSA bacteria) and clinical blood samples to be 100% specific and sensitive compared to qPCR results, and had 35 day stability under 25 °C storage. Furthermore, the color readout allows for quantitation of MRSA copies. Hence, this device is applicable for point-of-care MRSA detection.

Rapid and sensitive detection of tetracycline (TC) in the aquatic environment is important for the environmental ecosystem and human health. Herein, we presented on the first attempt to construct a fast‐responding gating system by using horseradish peroxidase (HRP)‐loaded functional RNA superstructures (HRP@3D RNA). Proteins were directly loaded into 3D RNA by the rolling circle transcription (RCT) reaction. We found that the specificity of aptamer‐ligand interaction could serve as a triggered manner to release the loaded HRP from 3D RNA. The above general scheme showed a few minutes of rapid release kinetics. According to the controlled‐release mechanism, we designed a disposable origami paper‐based analytical device (doPAD) coupled with HRP@3D RNA for colorimetric detection of TC. This doPAD can realize target recognition and signal readout by taking advantage of the unique spatial control advantages of 3D origami, thus generating a signal of color change that is positively related to the concentration of TC. This doPAD achieved the quantitative analysis of TC with a limit of detection of 10 nM within 41 min. We envision that the platform will be widely used in the field of environmental monitoring and biosensing. [ABSTRACT FROM AUTHOR]

Purpose – The purpose of this study was to develop an accurate, selective, low-cost and user-friendly colorimetric pad to detect formaldehyde at low concentration. Design/methodology/approach – 1-phenyl-1,3-butanedione, a reactive chemical, was selected to develop the colorimetric pad for indoor air formaldehyde measurement. Silica nanoparticle impregnated with the reactive chemical was coated on the cellulose filter surface to increase the reactive site. A certified formaldehyde permeation tube was used to generate six varied concentrations between 0.01 and 0.10 ppm in a test chamber. The color intensity on the pads was measured using an image processing program to produce a formaldehyde concentration reading chart. The colorimetric pad was tested for optimum reaction time, accuracy, precision, stability, selectivity and shelf life. Findings – The color of the pads changed from white to yellow and the color intensity varied with the concentrations and appeared to be stable after exposure to formaldehyde for 8 hours. At room temperature, the stability of the pad was 7 days, and shelf life was 120 days. The accuracy, precision and bias of the pad were 12.38%, 0.032 and 6.0%, respectively. Carbonyl compounds, benzene and toluene did not interfere with the reading of this developed colorimetric pad. Originality/value – The developed colorimetric pad meets NIOSH's criteria for an overall accuracy of ±25%, bias = 10%. They were accurate at low concentrations, user-friendly and had low cost compared to an electronic direct reading instrument (cost of chemicals and materials was 21.50 Bath or 0.69 USD per piece) so that favorable for the use of general people for health protection.

A paper-based analytical device (PAD) is presented with colorimetric/electrochemical dual readouts for the simultaneous sensing of total chromium (Cr) and hexavalent chromium (Cr(VI)). This device consists of a homemade three-electrode system and a patterned paper chip, integrating multiple functions including electrochemical detection, fluid driving, online oxidation, and colorimetric detection. The fiberglass filter paper with a hydrophilic microchannel was used to achieve self-driving fluidics without external equipment. One end of the microchannel was integrated with a homemade three-electrode system to achieve sample loading and electrochemical detection. The middle region on the microchannel was modified with oxidizing reagents to perform online pretreatment, and the yield of Cr(III) oxidation can reach 97.9%, ensuring reliable colorimetric detection of total Cr at another end of the microchannel modified with chromogenic agents. With this device, the signals of Cr(VI) (the signal peak at 0.29 V vs. Ag/AgCl) and total Cr can be obtained in one single injection. After optimization, the limit of detection (LOD) of Cr(VI) and total Cr were 0.01 mg L−1 and 0.06 mg L−1 and the linear ranges were 0.05–3.0 mg L−1 and 0.2–3.0 mg L−1, respectively. The relative standard deviations (RSD) of the electrochemical testing of Cr(VI) results were in a range 1.3%–8.7% (n = 3), and the RSD values of the colorimetric testing of total Cr were between 0.7–9.2% (n = 3). The device's reliability was demonstrated by performing the practical speciation of Cr in tap water, river water, and electroplating wastewater while the recoveries obtained using the present method were in the range 93.5–106%. Overall, the proposed device provides high application prospect in the on-site rapid Cr speciation. [ABSTRACT FROM AUTHOR]

The need to monitor the presence of cyanide (CN−) in water is necessary to minimize the risks to aquatic ecosystems and human health. In this paper, a paper-based analytical device (PAD) was fabricated by immobilizing silver nanoparticles (AgNPs) on filter paper (FP) for the semi-quantitative colorimetric detection of CN− in water. The average diameter of the synthesized AgNPs was estimated to be around 26.23 ± 8.37 nm, with a characteristic optical absorption peak around 420 nm. Scanning electron microscopy and energy-dispersive X-ray spectroscopy results confirmed the successful immobilization of AgNPs on the filter paper via direct immersion technique. The potential of the fabricated FP-AgNPs PAD as a colorimetric sensor for CN− was evaluated using water samples contaminated with various ions and CN− concentration. Here, a color change from yellow to colorless was instantly observed as the FP-AgNPs PAD was exposed to water samples containing CN−. Interestingly, no color change was observed for samples exposed to other analytes suggesting the good selectivity of the FP-AgNPs PAD. Ultraviolet–Visible spectroscopy results and digital image analysis revealed that the fabricated sensor can detect CN− with concentration down to 1.0 ppm. The colorimetric response was also obtained for real water samples spiked with CN−. The results stipulated in this work offer baseline information that can be used in developing highly selective and sensitive digital sensing devices for affordable, accessible, and fast water contaminant monitoring and other related applications. [ABSTRACT FROM AUTHOR]

Milk adulteration is life-threatening, and hence, its detection is an integral part of food safety. Most of the currently established detection techniques are expensive, sophisticated, and time-consuming. Though microfluidic devices offer a promising advantage in this regard, they are mostly colorimetric with sophisticated image processing, however, with compromised accuracy. In sharp contrast to this, we develop a low cost, simple yet accurate paper-based microfluidic device for the detection of starch concentration in milk. A 10 μl sample is injected in the inlet zone of the developed device and the resulting color transition length is captured using a smartphone after 5 min of the sample injection, and the starch concentration is measured by an in-house developed app, "starch-app". An excellent correlation (R2 = 0.9981) has been observed between the values of the starch concentration measured by the device and spectrophotometer in the range of 0–10% w/v. This novel, simple and inexpensive device has potential for its deployment in resource-constrained settings for estimation of starch adulteration in milk, especially in developing and under developing nations. [ABSTRACT FROM AUTHOR]

Herein, a paper-based glucose/air biofuel cell (BFC) was constructed and implemented for self-powered pesticide detection. Our developed paper-based chip relies on a hollow-channel to transport fluids rather than capillarity, which reduces analysis times as well as physical absorption. The gold nanoparticles (Au NPs) and carbon nanotubes (CNTs) were adapted to modify the paper fibers to fabricate the flexible conductive paper anode/cathode electrode (Au–PAE/CNT–PCE). Molecularly imprinted polymers (MIPs) using 2,4-dichlorophenoxyacetic acid (2,4-D) as a template were synthesized on Au–PAE for signal control. In the cathode, bilirubin oxidase (BOD) was used for the oxygen reduction reaction. Based on a competitive reaction between 2,4-D and glucose-oxidase-labeled 2,4-D (GOx-2,4-D), the amount of GOx immobilized on the bioanode can be simply tailored, thus a signal-off self-powered sensing platform was achieved for 2,4-D determination. Meanwhile, the coupling of the paper supercapacitor (PS) with the paper-based chip provides a simple route for signal amplification. Combined with a portable digital multi-meter detector, the amplified signal can be sensitively readout. Through rational design of the paper analytical device, the combination of BFC and PS provides a new prototype for constructing a low-cost, simple, portable, and sensitive self-powered biosensor lab-on-paper, which could be easily expanded in the field of clinical analysis and drug delivery. [ABSTRACT FROM AUTHOR]

A newly developed research topic, fabricated paper-based microfluidic sensors, was discussed in the field of low-cost environmental detection. Distinguished with the traditional dipstick or lateral-flow setups, these paper-based microfluidic sensors can serve as a tool for onsite quantitative and semi-quantitative measurements, without risks to cause environmental pollution. They have attracted increasing interest since the first easy-fabricated paper-based setup reported by Whitesides group in 2007. Most of the publications utilized paper-based sensors in clinical detection. In recent years, some groups started to use these sensors in environmental measurement, leading to precise, easy operation, low-cost, and eco-friendly methods for onsite detection. In this review, paper-based microfluidic sensors were briefly introduced, followed by literatures review and discussion for future perspectives. [ABSTRACT FROM AUTHOR]

In this work, indoxyl-glucoside was used as the substrate to develop a cost-effective, paper-based analytical device for the fluorescent and colorimetric dual-mode detection of β-glucosidase activity through a smartphone. The β-glucosidase can hydrolyze the colorless substrate indoxyl-glucoside to release indoxyl, which will be self-oxidized to generate green products in the presence of oxygen. Meanwhile, the green products emit bright blue-green fluorescence under ultraviolet–visible light irradiation at 365 nm. Fluorescent or colorimetric images were obtained by a smartphone, and the red-green-blue channels were analyzed by the Adobe Photoshop to quantify the β-glucosidase activity. Under the optimum conditions, the relative fluorescent and colorimetric signals have a good linear relationship with the activity of β-glucosidase, in the range of 0.01–1.00 U/mL and 0.25–5.00 U/mL, and the limits of detection are 0.005 U/mL and 0.0668 U/mL, respectively. The activities of β-glucosidase in a crude almond sample measured by the fluorescent and colorimetric methods were 23.62 ± 0.53 U/mL and 23.86 ± 0.25 U/mL, respectively. In addition, the spiked recoveries of normal human serum and crude almond samples were between 87.5% and 118.0%. In short, the paper-based device, combined with a smartphone, can provide a simple, environmentally friendly, and low-cost method for the fluorescent and colorimetric dual-mode detection of β-glucosidase activity. [ABSTRACT FROM AUTHOR]

[Display omitted] • The accurate measurement of creatinine (Ctn) in human biofluid samples is a challenge. • A stable carboxyl (C=O, COOH) groups were introduced on the electrode surface during electrochemical activation. • ePAD* was applied for the selective non-enzymatic detection of Ctn. • ePAD* detects Ctn in blood serum and urine samples with high reliability. • ePAD* is economic and accurate compared to existing methods for Ctn detection. Selective detection of creatinine (Ctn) is crucial for the clinical diagnosis of early-stage kidney dysfunction. A non-enzymatic electrochemical method for a highly selective and sensitive detection of Ctn in blood serum and urine samples was developed using an electrochemically activated carbon-coated paper-based analytical device (ePAD*). During activation, the growth of stable carboxyl (C = O, COOH) groups on the edge planes of the carbon surface was observed. The prepared ePAD* was characterized using various microscopic, spectroscopic, and electrochemical techniques. The ePAD* was utilized as a medium exchanger, and the voltammetric response of Ctn detection in the linear concentration from 0.03 to 45 mM with a detection limit of 5.41 µM was achieved. In addition, the selective detection of Ctn in the presence of potential interferences (i.e., uric acid, ascorbic acid, ammonia, and urea) was examined. The examination revealed a remarkable selectivity toward Ctn detection. Real-time detection of Ctn in blood serum and urine samples was performed. The results were correlated with the classical Jaffé colorimetric method. Our ePAD* exhibited a better electrochemical response. Thus, it is a highly potential diagnostic method for further development as a rapid and precise detection platform for point-of-care (POC) devices for examining patients with kidney disease. [ABSTRACT FROM AUTHOR]

Hydrogen peroxide is a key reagent in many analytical assays. At the same time, it is rather unstable and prone to evaporation. For these reasons, its application in sensors requiring reagents in solid state, for example in paper-based microfluidics, is hindered. Usually in paper-based analytical devices reagents are stored in a dried form within paper matrix until the device is used. This approach is not feasible in case of hydrogen peroxide. Here, hydrogen peroxide stabilization on paper with the aid of silica xerogel was studied and optimized to create long-term stable systems which rapidly deliver hydrogen peroxide. The variables affecting hydrogen peroxide stability such as gelation time, silica to H 2 O 2 ratio, type of solid support and storage conditions were optimized to find the combination of variables providing stable H 2 O 2 concentration for the longest time possible. Such paper-silica-H 2 O 2 composites allow to maintain steady hydrogen peroxide concentration for at least 27 days in the optimal conditions. Hydrogen peroxide is rapidly released from silica-paper matrix within a few minutes upon contact with water, without any byproducts. The obtained systems were characterized using scanning electron microscopy with energy dispersive spectroscopy and infrared spectroscopy, revealing that silica is present as a thin film covering cellulose fibers. Finally, to test the developed hydrogen peroxide stabilization method in real sensing scenario, a proof-of-concept paper-based sensor was created for phenolic content determination in fruits and wine. The outcome of this research will open new avenues in the development of user-friendly, long-term stable paper-based analytical devices which utilize hydrogen peroxide as one of reagents. Owing to the fact, that silica matrix is insoluble in water, the proposed H 2 O 2 stabilization method is compatible with most detection schemes without the risk of interfering with the assay. [Display omitted] • Hydrogen peroxide stabilization on paper with silica xerogel was studied. • Factors affecting hydrogen peroxide stability were optimized. • H 2 O 2 -silica-paper composites retain a stable H 2 O 2 amount for at least 27 days. • Paper-based analytical device for phenolic content determination was developed. [ABSTRACT FROM AUTHOR]

The separation of enantiomers plays a critical role in pharmaceutical development, ensuring therapeutic efficacy, safety, and patent protection. It enables the production of enantiopure drugs and enhances our understanding of the properties of chiral compounds. In this study, a straightforward and effective chiral detection strategy was developed for distinguishing between tryptophan (TRP) enantiomers. The approach involved the preparation of a magnetic molecularly imprinted chitosan (MMIC) for preparation of the sample, which was combined with a nitrocellulose membrane (a paper-based analytical device, PAD) integrated with D-TRP covalently grafted with polymethacrylic acid (PAD-PMA_D-TRP). Discriminating between the TRP enantiomers was achieved using AuNPs as a colorimetric probe. Indeed, the presence of D-TRP rapidly induced the aggregation of AuNPs due to its strong affinity to PAD-PMA_D-TRP, resulting in a noticeable change in the color of the AuNPs from red to purple. On the other hand, L-TRP did not induce any color changes. The chiral analysis could be easily performed with the naked eye and/or a smartphone. The developed method exhibited a detection limit of 3.3 µM, and it was successfully applied to detect TRP in serum samples, demonstrating good recovery rates. The proposed procedure is characterized by its simplicity, cost-effectiveness, rapidity, and ease of operation.

We report on the design of a simple all-DNA circuit with dual functions of signal amplification and signal reporting and its use for detection of human telomerase activity from cancer cells. The system utilizes a catalytic hairpin assembly (CHA) reaction for amplification, which produces split G-quadruplex outputs that assemble to form complete guanine quadruplex structures as reporting modules. As designed, a linear DNA sequence (the target) functions as a catalyst to drive cyclic programmed assembly of two hairpins, producing a DNA duplex with two guanine-rich sequences that assemble to form a complete Gq structure. The formation of the Gq element allows either fluorescence or colorimetric detection of the target. Examples are provided to demonstrate fluorescence detection of cancer cells' telomerase activities in solution and the first example of a CHAmodulated colorimetric assay for detecting telomerase activities of cancer cells using a simple paper device. [ABSTRACT FROM AUTHOR]

In this study, a paper-based sensor was developed for the detection of hydrogen-peroxide-related biomarkers, with glucose oxidase catalyzing as an example. Potassium iodide can catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine in the presence of hydrogen peroxide to colorize the paper-based biosensor detection area, which was imaged by a scanner, and the color intensity was analyzed by the Adobe Photoshop. Under the optimal conditions, the color intensity shows a good linear relationship with hydrogen peroxide and glucose concentrations in the ranges of 0.1–5.0 mM and 0.5–6.0 mM, respectively. The detection limit of hydrogen peroxide is 0.03 mM and the limit of quantification of glucose is 0.5 mM. Besides, the method was employed in measuring glucose concentration in fruit samples, and the spiked recoveries are in the range of 95.4–106.1%. This method is cost-effective, environmentally friendly, and easy to be operated, which is expected to realize the point-of-care testing of more hydrogen-peroxide-related biomarkers. [ABSTRACT FROM AUTHOR]

Here, a simple and portable paper‐based analytical device (PAD) based on the inherent capability of carbon quantum dots (CQDs) to serve as a great emitter for the bis(2,4,6‐trichlorophenyl)oxalate (TCPO)–hydrogen peroxide (H₂O₂) chemiluminescence (CL) reaction is introduced for the detection of harmful mercury ions (Hg2+). The energy is transferred from the unstable reaction intermediate (1,2‐dioxetanedione) to CQDs, as acceptors, and an intensive orange‐red CL emission is generated at ~600 nm, which is equal to the fluorescence emission wavelength of CQDs. The analytical applicability of this system was examined for the determination of Hg2+. It was observed that Hg2+ could significantly quench the produced emission, which can be attributed to the formation of a stable and nonluminescent Hg2+–CQDs complex. Accordingly, a simple and rapid PAD was established for monitoring Hg2+, with a limit of detection of 0.04 μg ml−1. No interfering effect on the signal was found from other examined cations, indicating the acceptable specificity of the method. The designed assay was appropriately utilized to detect Hg2+ ions in cosmetic samples with high efficiency. It was characterized by its low cost, ease of use, and was facile but accurate and high selective for the detection of Hg2+ ions. In addition, the portability of this probe makes it suitable for on‐site screening purposes. [ABSTRACT FROM AUTHOR]

The similar transmission patterns and early symptoms of respiratory viral infections, particularly severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza (H1N1), and respiratory syncytial virus (RSV), pose substantial challenges in the diagnosis, therapeutic management, and handling of these infectious diseases. Multiplexed point-of-care testing for detection is urgently needed for prompt and efficient disease management. Here, we introduce an electrochemical paper-based analytical device (ePAD) platform for multiplexed and label-free detection of SARS-CoV-2, H1N1, and RSV infection using immobilized pyrrolidinyl peptide nucleic acid probes. Hybridization between the probes and viral nucleic acid targets causes changes in the electrochemical response. The resulting sensor offers high sensitivity and low detection limits of 0.12, 0.35, and 0.36 pM for SARS-CoV-2 (N gene), H1N1, and RSV, respectively, without showing any cross-reactivities. The amplification-free detection of extracted RNA from 42 nasopharyngeal swab samples was successfully demonstrated and validated against reverse-transcription polymerase chain reaction (range of cycle threshold values: 17.43–25.89). The proposed platform showed excellent clinical sensitivity (100 %) and specificity (≥97 %) to achieve excellent agreement (κ ≥ 0.914) with the standard assay, thereby demonstrating its applicability for the screening and diagnosis of these respiratory diseases. [Display omitted] • The PNA-based ePAD was developed for multiplex detection of respiratory viruses. • This platform can be used to diagnose SARS-CoV-2, H1N1, and RSV infection. • The developed device achieves high specificity and sensitivity for the detection. • This system can be applied to clinical samples without requiring RNA amplification. • The analysis results corresponded well with those of the RT-PCR assays. [ABSTRACT FROM AUTHOR]

Branched-chain amino acids (BCAAs) are essential amino acids that play a crucial role in regulating animal physiology and metabolism. The lack of sensitive, rapid, and portable methods for measuring BCAA concentrations in blood samples hinders precise and dynamic monitoring of nutritional status during swine production. In this work, a paper-based analytical device (PAD) was developed for quantifying BCAAs in blood samples. The device utilized a redox reaction between BCAAs and NAD+ catalyzed by leucine dehydrogenase, producing NADH that could be detected by NADH-responsive polymer dots with high selectivity, sensitivity, and accuracy. A separation membrane was incorporated in the PAD for spontaneous blood-plasma separation. The fluorescent images were captured by using a smartphone. It required only 20 μL of blood volume, and the entire assay could be completed in less than 5 minutes. The quantitation range spanned from 35 μM to 1000 μM, covering the normal BCAA concentration range in porcine bloods. The BCAA concentration measured with the PAD assay was in good agreement with those obtained with an ELISA kit. In sum, this PAD-based BCAA assay offers a point-of-care testing approach for monitoring BCAA levels in pig production, especially with a low-protein diet. • A portable paper-based analytical device was developed for BCAAs quantification. • The NADH-sensitive polymer dots were utilized as the ratiometric fluorescent probe. • A ratiometric fluorescent assay was achieved by using a smartphone. • Total BCAA level in pig blood was accurately determined. [ABSTRACT FROM AUTHOR]

A novel and facile method by sewing low-cost fiber electrodes on paper is proposed to fabricate electrochemical paper-based analytical devices (ePADs). Prussian Blue (PB) and Ag/AgCl were deposited on carbon fibers to prepare the working and reference electrodes. The hydrophilic carbon fiber was employed as the counter electrode. All electrodes were sewed on the paper substrate. The linear voltammetry scanning (LSV) and physical characterization results showed that the physico-chemical characteristics of the prepared electrode were maintained after sewing. As a proof of concept, the ePAD fabricating by sewing was used to detect hydrogen peroxide (H 2 O 2). The limit of detection (LOD) achieved 0.9 µM and the ePAD possessed high anti-interference ability to a variety of common ions and biological molecules. Additionally, the robustness of the ePADs against bending (error 6.2 %) manifested that the ePAD had excellent practical applicability. The good reproducibility (3.65 %) displayed by the current responses confirmed that this strategy can be profitably adopted to easily assemble ePADs in a highly flexible manner. In addition, a user-friendly reagent-free ePAD was proposed to determine the content of H 2 O 2 in real water sample without treatment. The good recovery percentage (95.7–99.7 %) demonstrated the high feasibility of the reagent-free ePAD with facile carbon fiber-sewed electrodes in the detection of H 2 O 2 from complex water samples. These results confirm that the sewing method is feasible and promising. This work provides a new insight into fabrication of highly sensitive and highly selective ePADs with fiber electrodes. [Display omitted] [ABSTRACT FROM AUTHOR]

An instrument-free, user-friendly, and cost-effective PAD colorimetric sensor that can quantitative detection of H 2 O 2 has been realized by using bimetallic nickel–cobalt selenides as highly active peroxidase mimic and smartphone integrated with dark cavity as detector. Integrated with an ANN model and a self-compiled easy-to-use smartphone APP, the intelligent and on-site detection of H 2 O 2 was constructed, which exhibited an ultra-wide dynamic range. [Display omitted] • An effective paper-based colorimetric sensor was prepared based on nanozyme. • The optimized Ni 0.75 Co 0.25 Se exhibited excellent peoxidase-mimetic activities. • The on-site detection of H 2 O 2 was constructed by a self-compiled smartphone APP. • The machine learning-assisted sensor can detect H 2 O 2 over a wide dynamic range. Paper-based analytical devices (PADs) integrated with smartphones have shown great potential in various fields, but they also face challenges such as single signal reading, complex data processing and significant environmental impacting. In this study, a colorimetric PAD platform has been proposed using bimetallic nickel–cobalt selenides as highly active peroxidase mimic, smartphone with 3D-printing dark-cavity as a portable detector and an artificial neural network (ANN) model as multi-signal processing tool. Notably, the optimized nickel–cobalt selenides (Ni 0.75 Co 0.25 Se with Ni to Co ratio of 3/1) exhibit excellent peoxidase-mimetic activities and are capable of catalyzing the oxidation of four chromogenic reagents in the presence of H 2 O 2. Using a smartphone with image capture function as a friendly signal readout tool, the Ni 0.75 Co 0.25 Se based four channel colorimetric sensing paper is used for multi-signal quantitative analysis of H 2 O 2 by determining the Grey, red (R), green (G) and blue (B) channel values of the captured pictures. An intelligent on-site detection method for H 2 O 2 has been constructed by combining an ANN model and a self-programmed easy-to-use smartphone APP with a dynamic range of 5 μM to 2 M. Noteworthy, machine learning-assisted smartphone sensing devices based on nanozyme and 3D printing technology provide new insights and universal strategies for visual ultrasensitive detection in a variety of fields, including environments monitoring, biomedical diagnosis and safety screening. [ABSTRACT FROM AUTHOR]

The generation of DNase type I 3′OH DNA ends is closely related to the harm of endogenous reactive oxygen species (ROS) and environmental genotoxic agents. The evaluation of this type of DNA damage plays an important role in clinical intervention and environmental toxicity assessment. Terminal deoxynucleotidyl transferase (TdT)-assisted isothermal amplification (TAIA) offers a facile and versatile way to detect DNase type I 3′OH DNA ends. Its ability of templated-independent isothermal amplification is one unique feature. Here, we reported a paper-based analytical device (PAD) coupled with a smartphone for the detection of DNase type I 3′OH DNA ends using TAIA and colorimetric signal readout. We achieved the integration of cell lysis, DNA extraction, TAIA, horseradish peroxidase (HRP)-enabled colorimetric reaction, and signal readout. This device could achieve a limit of detection of 264 cells with a total assay time of less than 45 min. By combining PAD with a smartphone, the integrated platform could be used for the visual and quantitative analysis of DNA damages with the advantages of ease-to-use, fast response, inexpensive, and instrument free. Furthermore, successful assessment of the genotoxicity in wastewater effluents suggested the great promise of the integrated platform for on-site testing in practical applications.

Herein, a paper-based glucose/air biofuel cell (BFC) was constructed and implemented for self-powered pesticide detection. Our developed paper-based chip relies on a hollow-channel to transport fluids rather than capillarity, which reduces analysis times as well as physical absorption. The gold nanoparticles (Au NPs) and carbon nanotubes (CNTs) were adapted to modify the paper fibers to fabricate the flexible conductive paper anode/cathode electrode (Au–PAE/CNT–PCE). Molecularly imprinted polymers (MIPs) using 2,4-dichlorophenoxyacetic acid (2,4-D) as a template were synthesized on Au–PAE for signal control. In the cathode, bilirubin oxidase (BOD) was used for the oxygen reduction reaction. Based on a competitive reaction between 2,4-D and glucose-oxidase-labeled 2,4-D (GOx-2,4-D), the amount of GOx immobilized on the bioanode can be simply tailored, thus a signal-off self-powered sensing platform was achieved for 2,4-D determination. Meanwhile, the coupling of the paper supercapacitor (PS) with the paper-based chip provides a simple route for signal amplification. Combined with a portable digital multi-meter detector, the amplified signal can be sensitively readout. Through rational design of the paper analytical device, the combination of BFC and PS provides a new prototype for constructing a low-cost, simple, portable, and sensitive self-powered biosensor lab-on-paper, which could be easily expanded in the field of clinical analysis and drug delivery.

In this work, indoxyl-glucoside was used as the substrate to develop a cost-effective, paper-based analytical device for the fluorescent and colorimetric dual-mode detection of β-glucosidase activity through a smartphone. The β-glucosidase can hydrolyze the colorless substrate indoxyl-glucoside to release indoxyl, which will be self-oxidized to generate green products in the presence of oxygen. Meanwhile, the green products emit bright blue-green fluorescence under ultraviolet–visible light irradiation at 365 nm. Fluorescent or colorimetric images were obtained by a smartphone, and the red-green-blue channels were analyzed by the Adobe Photoshop to quantify the β-glucosidase activity. Under the optimum conditions, the relative fluorescent and colorimetric signals have a good linear relationship with the activity of β-glucosidase, in the range of 0.01–1.00 U/mL and 0.25–5.00 U/mL, and the limits of detection are 0.005 U/mL and 0.0668 U/mL, respectively. The activities of β-glucosidase in a crude almond sample measured by the fluorescent and colorimetric methods were 23.62 ± 0.53 U/mL and 23.86 ± 0.25 U/mL, respectively. In addition, the spiked recoveries of normal human serum and crude almond samples were between 87.5% and 118.0%. In short, the paper-based device, combined with a smartphone, can provide a simple, environmentally friendly, and low-cost method for the fluorescent and colorimetric dual-mode detection of β-glucosidase activity.

In this study, a paper-based sensor was developed for the detection of hydrogen-peroxide-related biomarkers, with glucose oxidase catalyzing as an example. Potassium iodide can catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine in the presence of hydrogen peroxide to colorize the paper-based biosensor detection area, which was imaged by a scanner, and the color intensity was analyzed by the Adobe Photoshop. Under the optimal conditions, the color intensity shows a good linear relationship with hydrogen peroxide and glucose concentrations in the ranges of 0.1–5.0 mM and 0.5–6.0 mM, respectively. The detection limit of hydrogen peroxide is 0.03 mM and the limit of quantification of glucose is 0.5 mM. Besides, the method was employed in measuring glucose concentration in fruit samples, and the spiked recoveries are in the range of 95.4–106.1%. This method is cost-effective, environmentally friendly, and easy to be operated, which is expected to realize the point-of-care testing of more hydrogen-peroxide-related biomarkers.

The paper-based device for allopurinol detection was developed by an enzymatic reaction. The concept is that allopurinol will act by blocking the xanthine oxidase enzyme to produce uric acid and H2O2, resulting in a color change from purple to pale purple until it is colorless. The concept was proofed by spectrophotometry and was applied in four types of filter paper including Whatman Qualitative No. 1, No. 4, and No. 6, and Whatman 1 chromatography. The application of this system was observed using ImageJ analysis. The performance of the paper reveals that each Whatman has different characteristics with LOD and LOQ range were 1.62-2.24 ppm and 4.91-6.80 ppm, respectively. The stability study shows that all of the paper was stable for 3 days both at room temperature (around 24°C-26°C) and 4°C. The presence of another drug like paracetamol did not prevent allopurinol detection. The real traditional medicine sample was also analyzed to confirm the applicability of this paper, and the result showed good agreement with spectrophotometry data. [ABSTRACT FROM AUTHOR]

A microcapillary grooved paper-based analytical device capable of dual-mode sensing (colorimetric and electrochemical detection) was demonstrated for analysis of viscous samples (e.g., human saliva). Herein, a hollow capillary channel was constructed via laser engraved micropatterning functions as a micropump to facilitate viscous fluidic transport, which would otherwise impede analysis on paper devices. Using salivary thiocyanate as a model analyte, the proposed device was found to exhibit a promising sensing ability on paper devices without the need for sample pretreatment or bulky instrumentation, as normally required in conventional methods used for saliva analysis. An extensive linear dynamic range covering detection of salivary thiocyanate for both high and trace level regimes (5 orders of magnitude working range) was collectively achieved using the dual-sensing modes. Under optimal conditions, the limit of detection was 6 μmol L−1 with a RSD of less than 5%. An excellent stability for the μpumpPAD was also observed for over 30 days. Real sample analysis using the proposed device was found to be in line with the standard chromatographic method. Benefitting from simple fabrication and operation, portability, disposability, low sample volume (20 μL), and low cost (< 1 USD), the μpumpPAD is an exceptional alternative tool for the detection of various biomarkers in saliva specimens. [ABSTRACT FROM AUTHOR]

• Surface molecularly imprinted polymer with selective recognition capability to butachlor was fabricated. • A colorimetric paper chip combined with the smartphone was developed to detect butachlor in mung bean. • The sensor has highly selective and sensitive to butachlor with LOD is 1.43 ng/g. • The sensor is simple, cheap, portable and widely used for fast detection. A microfluidic paper chip colorimetric detection system based on surface molecular imprinting of zinc ferrite nanoparticles was established, and the detection images were obtained by smartphone for gray value analysis and determination of butachlor. The best functional monomers and addition ratio were selected by quantum chemical simulation calculation, the properties of the prepared molecularly imprinted polymers were analyzed, and the detection conditions were optimized. The linear range, sensitivity, and selectivity of the method were evaluated. The results showed that under the optimum conditions, the concentration of 2–80 ng/g had a good linear relationship (R2 is 0.9953), the detection limit was 1.43 ng/g, the specificity was good, and the whole detection process did not exceed 20 min. The microfluidic paper chip was applied to detect butachlor in mung bean samples. The results showed that the recovery was 93.4–106.4 %, and the relative standard deviation was less than 5.6 %. [ABSTRACT FROM AUTHOR]

In the present study, an alternative platform for trace copper ions (Cu(II)) determination using a diamond electrode in combination with a disposable paper‐based analytical device (d‐PAD) has been proposed. First, the complexation between Cu(II) and 1,10‐phenanthroline ligand was adsorptively accumulated onto a paper that is directly in contact with the diamond electrode and measured via square wave anodic stripping voltammetry. Under the optimal experimental conditions, the peak current was proportional to the concentration of Cu(II) in the range of 0.4–70 ng mL−1, and the detection limit was found to be 0.1 ng mL−1. Most importantly, this platform can be successfully applied to detect trace Cu(II) in different water samples (drinking water, tap water, groundwater and river water) with satisfactory results. Thus, the proposed d‐PAD could be a highly efficient platform for trace Cu(II) determination in environmental samples. [ABSTRACT FROM AUTHOR]

We conducted electrochemical immunochromatography using a developed electrochemical immunochromatography platform. We used immunoglobulin G (IgG) as a representative protein. We also conducted enzyme-linked immunosorbent assay (ELISA) as a control test. We used the same reagents except for the substrate and compared their limits of detection (LODs) and detection ranges. We succeeded in measuring IgG concentrations ranging from 30 ng/ml to 10 μg/ml and obtained 23.00 ng/ml as the LOD for electrochemical immunochromatography. The detection signal was not saturated even at 10 μg/ml, so it seems that higher concentrations of IgG can also be measured. The LOD was approximately 80-fold higher than that of ELISA (0.30 ng/ml); however, the detection range was one order of magnitude higher than that of ELISA. While electrochemical immunochromatography is less sensitive than ELISA, we demonstrated sufficient sensitivity for its use in a point-of-care testing (POCT) device. Furthermore, this information will be very helpful when introducing an assay system already established in ELISA to POCT. [ABSTRACT FROM AUTHOR]

Colorimetric sandwich-type biosensors that can both provide sensitivity competitive with fluorescence-based approaches, and leverage reagents that are cost-effective, widely available and as safe as possible, are highly sought after. Herein, we demonstrate an alternative highly-sensitive colorimetric method for paper-based sandwich-type biosensing that uses starch–iodide complexation to simplify practical biosensing using ubiquitous reagents. Targeting the mycotoxin ochratoxin A (OTA), a covalently-immobilised OTA antibody on a cellulose surface captures OTA and forms a sandwich with OTA aptamer-conjugated glucose oxidase. Adding the chromogenic reagents at an optimized concentration, a distinct blue color develops within 30 min, offering excellent contrast with the clear/white of the negative sample. With a sampling volume down to just 5 μL, the assay exhibits concentration limits of detection and quantitation of 20 and 320 pg mL−1, respectively, and a linear range from 10−1 to 105 ng mL−1 (R2 = 0.997). The method displays excellent selectivity against related mycotoxins, excellent %recovery (95–117%) and robust operation in complex matrices (beer, urine and human serum), with no significant difference versus gold-standard liquid chromatography. Along with its excellent analytical performance, this assay benefits from non-toxic and extremely cheap reagents that can be safely disposed of in the field, and presents an attractive alternative to toxic dyes and nanoparticles. Image 1 • Novel colorimetric antibody-aptamer sandwich assay using starch-iodine complexation. • Central composite design for fast and efficient sensor optimization. • Ultrasensitive detection of OTA using PADs with LoD of 20 pg mL−1 using 5 μL. • Applied for OTA detection in beverages, urine and human serum. [ABSTRACT FROM AUTHOR]

Rapid and sensitive detection of acrylamide in food samples is important for food safety and public health. Here, we describe a disposable origami paper-based analytical device (denoted doPAD) for colorimetric detection of acrylamide. This device uniquely exploits 3D origami folding paper for spatial control of the target recognition and signal readout, thus resulting in a positive correlation between the signals and the analytes. Under optimal conditions, the device achieved the quantitative analysis of acrylamide with a limit of detection of 1.13 μg/L within 120 min (including a derivatization time of 90 min and an assay time of 21 min). Furthermore, our method allowed the rapid and sensitive detection of acrylamide in complex food matrices. We envision that the platform described will find useful applications in the fields of food safety and environmental health.

A point-of-care (POC) can be defined as an in vitro diagnostic test that can provide results within minutes. It has gained enormous attention as a promising tool for biomarkers detection and diagnosis, as well as for screening of chronic noncommunicable diseases such as diabetes mellitus. Diabetes mellitus type 2 is one of the metabolic disorders that has grown exponentially in recent years, becoming one of the greatest challenges to health systems. Early detection and accurate diagnosis of this disorder are essential to provide adequate treatments. However, efforts to reduce incidence should remain not only in these stages but in developing continuous monitoring strategies. Diabetes-monitoring tools must be accessible and affordable; thus, POC platforms are attractive, especially paper-based ones. Paper-based POCs are simple and portable, can use different matrixes, do not require highly trained staff, and are less expensive than other platforms. These advantages enhance the viability of its application in low-income countries and hard-to-reach zones. This review aims to present a critical summary of the main components required to create a sensitive and affordable enzymatic paper-based POC, as well as an oriented analysis to highlight the main limitations and challenges of current POC devices for diabetes type 2 monitoring and future research opportunities in the field.

Paper-based analytical devices (PADs) have encountered a wealth of applications in recent years thanks to the numerous advantages of paper as a support. A silver nanoflower (AgNF) modified paper-based dual substrate for both surface-enhanced Raman spectroscopy (SERS) and ambient pressure paper spray mass spectrometry (PS-MS) was developed. AgNFs were immobilized on nylon-coated paper modified with silver and ethylenediamine. The developed substrate was characterized via scanning electron microscopy and infrared spectroscopy. The densely packed nanoscale petals of the AgNFs lead to a large number of so-called hot spots at their overlapping points, which result in an enhancement of the Raman signal. In addition, the presence of the AgNFs produces an increase in the sensitivity of the mass spectrometric analysis as compared with bare paper and nylon/Ag-coated paper. The dual substrate was evaluated for the identification and quantification of ketoprofen in aqueous standards as well as human saliva from healthy volunteers. The method enables the determination of ketoprofen with a limit of detection and limit of quantification via PS-MS of 0.023 and 0.076 mg L−1, respectively, with a relative standard deviation (RSD) of 3.4% at a concentration of 0.1 mg L−1. This dual substrate enables the simple and fast detection of ketoprofen with minimal sample preparation, providing complementary Raman and mass spectrometric information. [ABSTRACT FROM AUTHOR]

A new paper-based analytical device based on bulk ion-selective optodes (ISOs) for dual Ag+ and Hg2+ detection has been developed. A plasticized PVC hydrophobic phase composed of 25,27-di(benzothiazolyl)-26,28-hydroxycalix[4]arene (CU1) as an ion-selective ionophore, potassium tetrakis(4-chlorophenyl)borate as an ion-exchanger and chromoionophore XIV as a lipophilic pH indicator was entrapped in the pores of cellulose paper. This paper strip showed higher selectivity for Ag+ and Hg2+ over common alkali, alkaline earth and some transition metal ions with a color change from blue to yellow. With the proposed sensor, Ag+ and Hg2+ can be measured with the range of 1.92 × 10−6 to 5.00 × 10−3 M for Ag+ and 5.74 × 10−7 to 5.00 × 10−5 M for Hg2+ with a limit of detection of 1.92 × 10−6 M for Ag+ and 5.74 × 10−7 M for Hg2+. The proposed sensor was successfully applied to determine the amount of mercury in various water sources and the amount of silver in cleaning product samples containing silver nanoparticles (AgNPs). The results were in good agreement with inductively couple plasma-optical emission spectrometric measurements (ICP-OES). Image 1 • Paper-based bulk ion-selective optode (ISOs) was fabricated. • Benzothiazole calix[4]arene was employed as an ionophore. • The sensor was used for dual colorimetric Ag+ and Hg2+ detection. • High selectivity for Ag+ and Hg2+ over other metal ions was achieved. [ABSTRACT FROM AUTHOR]

We developed a paper-based analytical device based on the electropolymerization of poly (3,4-ethylenedioxythipohene) (PEDOT) and graphene oxide (GO) composites on the ITO substrate for the detection of uric acid (UA) in authentic human saliva. Scanning electron microscopy, UV–vis spectrum and X–ray diffraction confirmed the formation of porous PEDOT combined with GO film during the electropolymerization process. The nanocomposite based sensor showed an enhanced electrocatalytic activity toward UA with high sensitivity and stability. We demonstrate that UA can be directly detected in undiluted saliva using the paper-based electroanalytical device with no interference from ascorbic acid and dopamine that are normally present in biological fluids. The results indicated that the developed device is promising for non-invasive monitoring of salivary UA in human body. A paper-based electroanalytical device integrated with a disposable ITO electrode based on PEDOT/GO nanocomposite was applied for the evaluation of salivary uric acid in authentic human saliva. Image 1 • A paper-based electroanalytical device was developed for the salivary uric acid analysis. • The PEDOT-GO nanocomposite based disposable sensor showed an enhanced electrocatalytic activity toward uric acid. • Salivary uric acid in human saliva can be directly detected with the sensor. • The sensor is low-cost, portable and promising for non-invasive monitoring of salivary uric acid. [ABSTRACT FROM AUTHOR]

This work presents a novel development that exploits the concept of in-situ gas-separation together with a specific enzymatic colorimetric detection to produce a portable biosensor called "Blood Alcohol Micro-pad" for direct quantitation of ethanol in whole blood. The thin square device (25 mm × 25 mm × 1.8 mm) comprises two layers of patterned filter paper held together with a double-sided mounting tape with an 8-mm circular hole (the headspace). In operation, the reagent is deposited on one layer and covered with sticky tape. Then 8 μL of a blood sample is dispensed onto the opposite layer and covered with sticky tape. Diffusion of ethanol across the 1.6 mm narrow headspace permits selective detection of ethanol by the enzymatic reagents deposited on the opposite layer. This reagent zone contains alcohol oxidase, horseradish peroxidase and 2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, as the chromogenic reagent. The color intensity, measured from the recorded digital image, resulting from the enzymatic assay of ethanol, correlates with the concentration of blood alcohol. The results obtained with spiked mice and sheep blood samples, using an external calibration in the range of 1–120 mg dL−1ethanol, gave recoveries of 93.2–104.4% (n = 12). The "Blood Alcohol Micro-pad" gave good precision with %RSD <1 (50 mg dL−1 ethanol, n = 10) and limit of quantification (10SD of intercept/slope) of 11.56 mg dL−1. The method was successfully validated against a headspace gas chromatography-mass spectrometric method. It has good potential for development as a simple and convenient blood alcohol sensor for on-site testing. Image 1 • The first paper-based biosensor for quantifying alcohol content directly from whole blood. • In-situ headspace separation with colorimetric enzymatic assay for rapid readout within 5 min. • The device is portable, low cost and disposable. • Results are promising for roadside check and toxicological test. [ABSTRACT FROM AUTHOR]

Herein we report the first two distance-based microfluidic paper-based analytical devices (μPADs) using fluorescence to quantify aluminum. In addition to their read-by-eye quantification, the devices are simple to fabricate, require no sample pretreatment or preconcentration, and have a shelf life of >5 months. The first device is designed in a "chemometer" format where the length of a fluorescent band linearly responds to an Al(III) concentration. The second device uses a radial design where the fluorescent diameter also linearly responds to an Al(III) concentration. The chemometer device has a detection limit of 2.5 ppm (100 μM) and a linear range from 2 to 54 ppm Al(III) (100 μM–1 mM), R2 = 0.989). The radial device has a detection limit of 0.9 ppm (33 μM) and a linear range from 2 to 24 ppm Al(III) (100–900 μM, R2 = 0.968). The utility of the μPADs were successfully demonstrated by measuring Al(III) in two water effluent samples from the Gold King Mine near Silverton, CO. Image 1 • Distance-based detection of Al(III) in water samples. • Radial and linear distance based detection demonstrated. • Method gives accurate results in natural waters. [ABSTRACT FROM AUTHOR]

A paper-based analytical device (PAD) with an integrated composite electrode for non-enzymatic creatinine sensing was developed. The electrode was produced and optimization was efficiently accomplished using a rapid digital dispensing approach. The electrochemical sensor was fabricated using an HP D300 digital dispenser to deliver a copper oxide and ionic liquid composite onto an electrochemically reduced graphene modified screen-printed carbon electrode (CuO/IL/ERGO/SPCE) on a PAD. The modified electrode was characterized using electrochemical and microscopic techniques. Electrochemical detection of creatinine was performed on the SPCE using amperometry at a constant potential. Under optimized conditions, the paper-based sensor exhibited a linear range of 0.01–2.0 mM (R2 = 0.99) and the limit of detection was 0.22 μM (S/N = 3, IUPAC definition) for creatinine. The simple fabrication process, low cost, and clinically appropriate creatinine sensitivity make this device applicable for point-of-care use. Image 1 • A CuO/IL/ERGO modified paper-based screen printed electrode was constructed and used as a non-enzymatic creatinine sensor. • An HP D300 dispenser was used to enable efficient electrode modification, leading to improved sensitivity and reproducibility. • The sensitivity was sufficient for detecting clinically relevant creatinine levels in acute kidney injury diagnosis. [ABSTRACT FROM AUTHOR]

Field-amplified stacking (FAS) is a commonly used method for enhancing the sensitivity of charged species from low conductive media in capillary electrophoresis. FAS also showed significant sensitivity enhancement effect on a uniform paper fluidic channel by proper design of the electrolyte. In this paper, a novel method of introducing electric field gradient is proposed by geometry design of a 2D paper fluidic channel, and field amplification effect was successfully demonstrated with reduced requirement on the sample's conductivity. Sensitive colorimetric detection of microalbuminuria (MAU) from urine samples was demonstrated by mobile phone camera. Experimental results showed that, with active electric field motivation, up to 93.5% of the loaded protein probe could be effectively transferred and stacked into a narrow band on the newly designed paper fluidic channel. A limit of detection (LOD) of 6.5 mg‧L−1 HSA was achieved with a dynamic range of 10–300 mg‧L−1 (linear in the range of 10–100 mg‧L−1, R2 = 0.991). Combined with selective staining of albumin with bromophenol blue (BPB), the established method was applied to the detection of MAU from clinical urine samples, and consistent results with that of the clinical method were obtained. With this paper-based analytical device (PAD), MAU from highly conductive urine samples can be directly loaded and detected without any pretreatment. This method provides a way to develop highly sensitive point-of-care test (POCT) for rapid screening of some diseases. Image 1 • Novel field amplification achieved mainly by the change of paper channel geometry, rather than the conductivity difference. • Protein probe was actively transported with a high transfer rate up to 93.5%, vs. less than 50% only by capillary force. • Sensitivity was enhanced by 2 orders of magnitude as demonstrated by colorimetric detection of human serum albumin (HSA). • Detection of microalbuminuria (MAU) from clinical urine samples was successfully demonstrated. [ABSTRACT FROM AUTHOR]

[Display omitted] • A mix-and-measure fluorimetric method was developed for naftazone. • Naftazone quenches the fluorescence of GQDs via inner filter effect. • The method showed nano molar sensitivity for naftazone down to 40 nM. • Successful determination of naftazone in tablets and on equipment surfaces. • Smartphone detection of naftazone by PAD using UV-torch as an excitation source. A simple, fast, and direct mix-and-read spectrofluorimetric method has been developed for the sensitive determination of naftazone (NFZ) utilizing graphene quantum dots (GQDs) as a greener and highly luminescent nanosensor. NFZ effectively quenches the strong fluorescence of GQDs at λ ex /λ em of 350/440 nm via the inner filter effect mechanism. The nanosensor exhibits excellent linearity for NFZ over the concentration range of 0.46 to 186 μM with a limit of detection of 0.04 μM. The proposed method was validated for the successful determination of NFZ in tablets and on manufacturing equipment surfaces with good % recoveries of 98.4–101.6 and 96.3 – 102.2%, respectively. Furthermore, an integrated smartphone-based reader has been implemented and successfully applied for the determination of NFZ. The smartphone-based reader consists of a 365 nm UV torch as an excitation source, a smartphone for recording images, and smartphone-powered image analysis software for signal interpretation, together with a paper-based analytical device (PAD) utilizing filter paper as a substrate and correction fluid as a barrier for creation of detection zones. This smart platform showed excellent sensitivity with a limit of detection down to 0.12 nmol/zone, and it could be used for in-site determination of NFZ, especially for the limited resources laboratories. [ABSTRACT FROM AUTHOR]

Cyanide anion can be found in foodstuffs, tobacco smoke and a variety of types of waters, mainly originating from anthropogenic activities. Due to its highly toxic nature, several agencies have established limits for cyanide levels in water. Additionally, monitoring cyanide levels in biological samples, such as blood and urine, is crucial for obtaining clinical information about the health condition of patients. Therefore, there is a pressing need for the development of simple, cost-effective, and reliable analytical methods capable of quantifying cyanide at low concentrations. This study presents a novel analytical method for the selective and sensitive determination of cyanide based on analyte volatilization, pre-concentration via single-drop microextraction (SDME) using a selective reagent, and colorimetric quantification using a paper-based analytical device. For this, 10 mL of a liquid sample was acidified with phosphoric acid and the generated HCN was collected using a single drop of 3 μL of a palladium dimethylglyoximate solution (Pd (DMG) 2 2−) positioned in the flask headspace using a syringe. The reaction of Pd (DMG) 2 2− leads to the formation of Pd(CN) 4 2− and the demasking of the organic ligand. After 15 min of extraction time, the reagent drop was added to a paper-based analytical device that has been previously impregnated with 3 μL of nickel chloride, resulting in the formation of a red precipitate of nickel (II) dimethylglyoximate. Digital images of the paper-based device were captured and the red channel (R) was used for quantification purposes. Under optimized conditions, the method demonstrates a suitable linear relation (r2 > 0.99) ranging from 26 to 286 μg L−1 and a limit of detection of 5 μg L−1. As a proof of concept, cyanide levels were quantified in water and urine samples using this method. The proposed approach offers high sensitivity and selectivity while requiring only a small volume of reagents. Furthermore, it exhibits a high degree of portability for in-situ applications. A selective and sensitive method for the determination of cyanide using HS-SDEM. [Display omitted] • Cyanide in liquid samples was determined by combining HS-SDME and digital image analysis. • Colorimetric reaction is performed on a microfluidic paper-based analytical device. • The reagent is easy to prepare and highly selective for cyanide. • Under optimized conditions, a limit of detection of 5 μg L-1 was achieved. • The point-of-care device is suitable for cyanide determination in water and urine samples. [ABSTRACT FROM AUTHOR]

Severe antimicrobial resistance calls for developing rapid, sensitive and affordable methodological platform for clinical diagnosis of bacterial infection. Herein, a paper-based analytical device (PAD) for fluorescent (FL) detection and antimicrobial susceptibility test (AST) of Pseudomonas aeruginosa (P. aeruginosa) was fabricated by defining 96 hydrophilic microzones on a filter paper using a handheld stamp dipped with liquid wax. The size of microzones was designed to be consistent with traditional 96-well microplate, thus the FL signals can be collected by a commercialized microplate reader. Streptavidin was immobilized into the microzones by chitosan-glutaraldehyde crosslinking reaction, and then biotinylated bacteriophage tail fiber protein (TFP) was conjugated through biotin-streptavidin affinity system. TFP and fluorescein isothiocyanate (FITC) labeled antimicrobial peptide were used as capture agent and signal probe, respectively, for FL detection of P. aeruginosa on the PAD. The linear range for quantifying P. aeruginosa is 1.0 × 103 CFU/mL to 1.0 × 107 CFU/mL, with a detection limit of 137 CFU/mL. The PAD was also applied to conduct AST of P. aeruginosa for imipenem, meropenem, cefepime, amikacin, and gentamicin, and the results are consistent with the traditional broth dilution method. The PAD provides an affordable diagnosis platform for bacterial infection, especially in resource-limited institutes and countries. [ABSTRACT FROM AUTHOR]

Paper-based analytical devices (PADs) and Electrospun Fiber-Based Biosensors (EFBs) have aroused the interest of the academy and industry due to their affordability, sensitivity, ease of use, robustness, being equipment-free, and deliverability to end-users. These features make them suitable to face the need for point-of-care (POC) diagnostics, monitoring, environmental, and quality food control applications. Our work introduces new and experienced researchers in the field to a practical guide for fibrous-based biosensors fabrication with insight into the chemical and physical interaction of fibrous materials with a wide variety of materials for functionalization and biofunctionalization purposes. This research also allows readers to compare classical and novel materials, fabrication techniques, immobilization methods, signal transduction, and readout. Moreover, the examined classical and alternative mathematical models provide a powerful tool for bioanalytical device designing for the multiple steps required in biosensing platforms. Finally, we aimed this research to comprise the current state of PADs and EFBs research and their future direction to offer the reader a full insight on this topic.