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The aim of this study was to identify the nature and cause of foxing spots in a historical manuscript. This manuscript was a Holy Quran from the beginning of the Qajar period and the end of the 18th century. Samples were incubated for 14 days and were evaluated for the presence of fungal activity. UV fluorescence photography, micro X-ray fluorescence spectroscopy and Fourier transform infrared spectroscopy were also used to investigate the characteristics and causes of foxing spots. The results showed that there was no fungal activity in the foxing spots of this manuscript. Based on the morphology of the stain in UV fluorescence photography, these foxing stains are of the Bullseye type, usually associated with metal ions. µXRF spectroscopy also showed a high accumulation of iron and copper at the site of these spots. This indicates abiotic foxing in this manuscript. Based on FTIR spectroscopy and peak deconvolution and fitting by Gaussian function, abiotic foxing increases the cellulose oxidation rate. Intensification of cellulose oxidation in foxing stains can be considered as one of the reasons for paper discoloration.

Paper spray ionization (PSI) is an extractive ambient ionization technique for mass spectrometry (MS), whereby a triangular paper tip serves as the sampling base and the electrospray tip. During PSI, analytes are extracted and transported to the edge of the paper tip by the applied spraying solvent. Analytes can be purified from a sample matrix and separated from each other by this transportation process. In this study, we investigated and utilized the analyte transportation process of PSI for the in situ separation and analysis of lipid mixtures. We found that differential transport of phosphatidylcholine (PC) and triacylglycerol (TAG), the two most abundant lipid classes in animals, occurred during PSI. We also found that the order in which these lipids moved strongly depended on how the spraying solvent was applied to the paper base. The more polar PC moved faster than the less polar TAG during PSI, when a polar solvent was slowly fed into a paper tip, whereas TAG was transported faster than PC when excess solvent was applied to the tip at once. In addition, we achieved a complete separation and detection of PC and TAG by slowly supplying a nonpolar solvent to a PSI tip.

Over the past decades, paper-based lateral flow immunoassays (LFIAs) have been extensively developed for rapid, facile, and low-cost detection of a wide array of target analytes in a point-of-care manner. Conventional home pregnancy tests are the most significant example of LFAs, which detect elevated concentrations of human chorionic gonadotrophin (hCG) in body fluids to identify early pregnancy. In this work, we have upgraded these platforms to a higher version by developing a customized microfluidic paper-based analytical device (μPAD), as the new generation of paper-based point-of-care platforms, for colorimetric immunosensing. This will offer a cost-efficient and environmentally friendly alternative platform for paper-based immunosensing, eliminating the need for nitrocellulose (NC) membrane as the substrate material. The performance of the developed platform is demonstrated by detection of hCG (as a model case) in urine samples and subsequently indicating positive or negative pregnancy. A dual-functional silane-based composite was used to treat filter paper in order to enhance the colorimetric signal intensity in the detection zones of μPADs. In addition, microfluidic pathways were designed in a manner to provide the desired regulated fluid flow, generating sufficient incubation time (delays) at the designated detection zones, and consequently enhancing the obtained signal intensity. The presented approaches allow to overcome the existing limitations of μPADs in immunosensing and will broaden their applicability to a wider range of assays. Although, the application of the developed hCG μPAD assay is mainly in qualitative (i.e., positive or negative) detection of pregnancy, the semi-quantitative measurement of hCG was also investigated, indicating the viability of this assay for sensitive detection of the target hCG analyte within the related physiological range (i.e., 10–500 ng/mL) with a LOD value down to 10 ng/mL.

In recent years, microfluidic lab-on-paper devices have emerged as a rapid and low-cost alternative to traditional laboratory tests. Additionally, they were widely considered as a promising solution for point-of-care testing (POCT) at home or regions that lack medical infrastructure and resources. This review describes important advances in microfluidic lab-on-paper diagnostics for human health monitoring and disease diagnosis over the past five years. The review commenced by explaining the choice of paper, fabrication methods, and detection techniques to realize microfluidic lab-on-paper devices. Then, the sample pretreatment procedure used to improve the detection performance of lab-on-paper devices was introduced. Furthermore, an in-depth review of lab-on-paper devices for disease measurement based on an analysis of urine samples was presented. The review concludes with the potential challenges that the future development of commercial microfluidic lab-on-paper platforms for human disease detection would face.

The development of diagnostic tools for measuring a wide spectrum of target analytes, from biomarkers to other biochemical parameters in biological fluids, has experienced a significant growth in the last decades, with a good number of such tools entering the market. Recently, a clear focus has been put on miniaturized wearable devices, which offer powerful capabilities for real-time and continuous analysis of biofluids, mainly sweat, and can be used in athletics, consumer wellness, military, and healthcare applications. Sweat is an attractive biofluid in which different biomarkers could be noninvasively measured to provide rapid information about the physical state of an individual. Wearable devices reported so far often provide discrete (single) measurements of the target analytes, most of them in the form of a yes/no qualitative response. However, quantitative biomarker analysis over certain periods of time is highly demanded for many applications such as the practice of sports or the precise control of the patient status in hospital settings. For this, a feasible combination of fluidic elements and sensor architectures has been sought. In this regard, this paper shows a concise overview of analytical tools based on the use of capillary-driven fluidics taking place on paper or fabric devices integrated with solid-state sensors fabricated by thick film technologies. The main advantages and limitations of the current technologies are pointed out together with the progress towards the development of functional devices. Those approaches reported in the last decade are examined in detail.

The fabrication of low-cost, flexible, and recyclable electronic devices has been the focus of many research groups, particularly for integration in wearable technology and the Internet of Things (IoT). In this work, porous zinc oxide (ZnO) nanostructures are incorporated as a UV sensing material into the composition of a sustainable water-based screen-printable ink composed of car-boxymethyl cellulose (CMC). The formulated ink is used to fabricate flexible and foldable UV sensors on ubiquitous office paper. The screen-printed CMC/ZnO UV sensors operate under low voltage (≤2 V) and reveal a stable response over several on/off cycles of UV light exposure. The devices reach a response current of 1.34 ± 0.15 mA and a rise and fall time of 8.2 ± 1.0 and 22.0 ± 2.3 s, respectively. The responsivity of the sensor is 432 ± 48 mA W−1, which is the highest value reported in the literature for ZnO-based UV sensors on paper substrates. The UV-responsive devices display impressive mechanical endurance under folding, showing a decrease in responsivity of only 21% after being folded 1000 times. Their low-voltage operation and extreme folding stability indicate a bright future for low-cost and sustainable flexible electronics, showing potential for low-power wearable applications and smart packaging. publishersversion published

Currently, there is an increasing interest in the use of biopolymers in industrial applications to replace petroleum-based additives, since they are abundantly available, renewable and sustainable. Cottonseed protein is a biopolymer that, when used as a modifier, has shown improved performance for wood adhesives and paper products. Thus, it would be useful to explore the feasibility of using cellulose nanomaterials to further improve the performance of cottonseed protein as a paper strength agent. This research characterized the performance of cottonseed protein isolate with/without cellulose nanofibers (CNFs) and cellulose nanocrystals (CNCs) to increase the dry strength of filter paper. An application of 10% protein solution with CNCs (10:1) or CNFs (50:1) improved the elongation at break, tensile strength and modulus of treated paper products compared to the improved performance of cottonseed protein alone. Further analysis using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) indicated that the cottonseed protein/nanocellulose composites interacted with the filter paper fibers, imparting an increased dry strength.

Point-of-care monitoring of hydrogen peroxide is important due to its wide usage in biomedicine, the household and industry. Herein, a paper sensor is developed for sensitive, visual and selective detection of H2O2 using a mesoporous metal oxide hollow sphere as a nanozyme. The mesoporous CuO hollow sphere is synthesized by direct decomposition of copper–polyphenol colloidal spheres. The obtained mesoporous CuO hollow sphere shows a large specific surface area (58.77 m2/g), pore volume (0.56 cm3/g), accessible mesopores (5.8 nm), a hollow structure and a uniform diameter (~100 nm). Furthermore, they are proven to show excellent peroxidase-like activities with Km and Vmax values of 120 mM and 1.396 × 10−5 M·s−1, respectively. Such mesoporous CuO hollow spheres are then loaded on the low-cost and disposable filter paper test strip. The obtained paper sensor can be effectively used for detection of H2O2 in the range of 2.4–150 μM. This work provides a new kind of paper sensor fabricated from a mesoporous metal oxide hollow sphere nanozyme. These sensors could be potentially used in bioanalysis, food security and environmental protection.

The red imported fire ant (RIFA, Solenopsis invicta) is an exotic aggressive pest that is notorious for its ability to seriously harm humans and animals, cause economic loss to agriculture, and damage ecosystems. This is the first study to validate the capability of filter paper adsorption as a feasible odor bearer of RIFAs and evaluate its use in detection dog training. Two live RIFA-experienced detection dogs achieved a mean 92% positive indication rate (PIR) on RIFA-scented papers with a relatively low false response rate (0.8%). The similar accuracies in recognizing live RIFAs (96%) and scented papers (92%) suggest that a filter paper is an effective odor reservoir. After training with live RIFA and scented filter papers, both RIFA-experienced and inexperienced detection dogs successfully indicated filter papers that were scented with at least 10 RIFAs for 4 h with a high PIR (&gt, 93%) and low false response rate (2%). Detection dogs correctly recognized the filter papers scented by 10 RIFAs for 24 h with a 97.6% PIR. Even for scented samples stored at −20 °C and 4 °C for 13 weeks, the positive indication rates (PIRs) were as high as 90%. These results suggest that filter paper is an effective RIFA odor bearer, and the scent can be maintained at least 13 weeks for dog identification. After RIFA-scented paper training, detection dogs showed high (&gt, 95%) PIRs for both RIFA-scented paper and live RIFAs and also successfully performed field studies. Using filter paper as a RIFA odor bearer is an effective and economical method for detection dog training and RIFA identification.

Since the introduction of paper-based analytical devices as potential diagnostic platforms a few decades ago, huge efforts have been made in this field to develop systems suitable for meeting the requirements for the point-of-care (POC) approach. Considerable progress has been achieved in the adaptation of existing analysis methods to a paper-based format, especially considering the chemiluminescent (CL)-immunoassays-based techniques. The implementation of biospecific assays with CL detection and paper-based technology represents an ideal solution for the development of portable analytical devices for on-site applications, since the peculiarities of these features create a unique combination for fitting the POC purposes. Despite this, the scientific production is not paralleled by the diffusion of such devices into everyday life. This review aims to highlight the open issues that are responsible for this discrepancy and to find the aspects that require a focused and targeted research to make these methods really applicable in routine analysis.

This work reports a modification of a fibrous cellulose material (paper) by the addition of polyacrylonitrile (PAN) fibres doped with 10,12–pentacosadiynoic acid (PDA). The fibres are sensitive to ultraviolet (UV) light. When the paper containing PAN–PDA is irradiated with UV light it changes colour to blue as a consequence of interaction of the light with PDA. The colour intensity is related to the absorbed dose, content of PAN–PDA fibres in the paper and the wavelength of UV radiation. The features of the paper are summarised after reflectance spectrophotometry and scanning microscopy analyses. All the properties of the modified paper were tested in accordance with adequate ISO standards. Moreover, a unique method for assessing the unevenness of the paper surface and the quality of printing was proposed by using a Python script (RGBreader) for the analysis of RGB colour channels. The modification applied to the paper can serve as a paper security system. The modified paper can act also as a UV radiation indicator.

Here, we designed the composition of the coating of the paper sheets composed of chitosan, bacterial cellulose (nanofibres), and ZnO with boosted antibacterial and mechanical activity. We investigated the compositions with ZnO exhibiting two different sizes/shapes: (1) rods and (2) irregular sphere-like particles. The proposed processing of bacterial cellulose resulted in the formation of nanofibers. Antimicrobial behavior was tested using E. coli ATCC® 25922™ following ASTM E2149-13a standard. Mechanical properties of the paper sheets were measured by comparison of tearing resistance, tensile strength, and bursting strength according to ISO 5270 standard. The increased antibacterial response is assigned to the combination of chitosan and ZnO (independently of its shape and size), while the boosted mechanical behavior is due to bacterial cellulose nanofibers. Therefore, the proposed composition is an interesting multifunctional mixture for coatings in food packaging applications.

Despite the potential in fabrication of microfluidic paper-based analytical devices (μPADs) for point-of-care testing (POCT) kits, the development of simple, accurate, and rapid devices with higher sensitivity remains challenging. Here, we report a novel method for 3D-μPAD fabrication with enclosed channels using vat photopolymerization to avoid fluid evaporation. In detail, height of the enclosed channels was adjusted from 0.3 to 0.17 mm by varying the UV exposure time from 1 to 4 s for the top barrier, whereas the exposure time for the bottom and side barriers was fixed. As a result, sample flow in the enclosed channels of 3D-μPADs showed lesser wicking speed with very scant evaporation compared to that in the hemi channels in the 3D-μPADs. The stoppage of evaporation in the enclosed channels significantly improved the gray intensity and uniformity in the detection zone of the 3D-μPADs, resulting in as low as 0.3 mM glucose detection. Thus 3D-μPADs with enclosed channels showed enhanced sensitivity compared to the 3D-μPADs with hemi channels when dealing with a small volume sample. Our work provides a new insight into 3D-μPAD design with enclosed channels, which redefines the methodology in 3D printing.

This review summarizes the state of the art of paper-based biosensors (PBBs) for coronavirus disease 2019 (COVID-19) detection. Three categories of PBB are currently being been used for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnostics, namely for viral gene, viral antigen and antibody detection. The characteristics, the analytical performance, the advantages and drawbacks of each type of biosensor are highlighted and compared with traditional methods. It is hoped that this review will be useful for scientists for the development of novel PBB platforms with enhanced performance for helping to contain the COVID-19 outbreak, by allowing early diagnosis at the point of care (POC).

Cellulose paper has been functionalized by nanoparticles such as Ag nanoparticles, TiO2, and BaTiO3 for versatile applications including supercapacitor, sensors, photoactivity, and packaging. Herein, zinc oxide (ZnO) nanosheet-modified paper (ZnO@paper) with excellent antibacterial properties was fabricated via a mild ZnCl2-urea eutectic solvent. In this proposed method, cellulose fibers as the raw material for ZnO@paper were treated by an aqueous solvent of ZnCl2-urea, the crystalline region was destroyed and [ZnCl]+-based cations were adsorbed on the surface of cellulose fibers, facilitating more ZnO growth on ZnO@paper. A flexible paper-based triboelectric nanogenerator (P-TENG) was made of ZnO@paper paired with a PTFE film. The P-TENG presents high triboelectric output performance and antibacterial activity. For instance, the output voltage and current of the P-TENG were 77 V and 0.17 μA, respectively. ZnO@paper showed excellent antibacterial activity against E. coli and S. aureus, suggesting that a P-TENG can restrain and kill the bacteria during the working process. The results also indicated that ZnO could improve the surface roughness of cellulose paper, enhancing the output performance of a flexible P-TENG. In addition, the potential application of a P-TENG-based pressure sensor for determining human motion information was also reported. This study not only produced a high-performance P-TENG for fabricating green and sustainable electronics, but also provides an effective and novel method for ZnO@paper preparation.

The use of fully printed electrochemical devices has gained more attention for the monitoring of clinical, food, and environmental analytes due to their low cost, great reproducibility, and versatility characteristics, serving as an important technology for commercial application. Therefore, a paper-based inkjet-printed electrochemical system is proposed as a cost-effective analytical detection tool for paraquat. Chromatographic paper was used as the printing substrate due its sustainable and disposable characteristics, and an inkjet-printing system deposited the conductive silver ink with no further modification on the paper surface, providing a three-electrode system. The printed electrodes were characterized with scanning electron microscopy, cyclic voltammetry, and chronopotentiometry. The proposed sensor exhibited a large surface area, providing a powerful tool for paraquat detection due to its higher analytical signal. For the detection of paraquat, square-wave voltammetry was used, and the results showed a linear response range of 3.0–100 μM and a detection limit of 0.80 µM, along with the high repeatability and disposability of the sensor. The prepared sensors were also sufficiently selective against interference, and high accuracy (recovery range = 96.7–113%) was obtained when applied to samples (water, human serum, and orange juice), showing the promising applicability of fully printed electrodes for electrochemical monitoring.

An origami three-dimensional design of a paper-based potentiometric sensor is described. In its simplest form, this electrochemical paper-based analytical device (ePAD) is made from three small parts of the paper. Paper layers are folded on each other for the integration of a solid contact ion selective electrode (here a carbon-paste composite electrode) and a solid-state pseudo-reference electrode (here writing pencil 6B on the paper), which are in contact with a hydrophilic channel fabricated on the middle part (third part) of the paper. In this case, the pseudo-reference and working electrodes are connected to the two sides of the hydrophilic channel and hence the distance between them is as low as the width of paper. The unmodified carbon paste electrode (UCPE) and modification with the crown ether benzo15-crown-5 (B15C5) represented a very high sensitivity to Cu (II) and Cd2+ ions, respectively. The sensor responded to H2O2 using MnO2-doped carbon paste electrode (CPE). Furthermore, a biosensor was achieved by the addition of glucose oxidase to the MnO2-doped CPE and hence made it selective to glucose with ultra-sensitivity. In addition to very high sensitivity, our device benefits from consuming a very low volume of sample (10.0 µL) and automatic sampling without need for sampling devices.

This paper proposes a combined strategy of using paper-based competitive immunochromatography and a near field communication (NFC) tag for wireless cotinine determination. The glucose oxidase labeled cotinine antibody specifically binds free cotinine in a sample, whereas the unoccupied antibody attached to BSA-cotinine at the test line on a lateral flow strip. The glucose oxidase on the strip and an assistant pad in the presence of glucose generated H2O2 and imposed the Ag oxidation on the modified electrode. This enabled monitoring of immunoreaction by either electrochemical measurement or wireless detection. Wireless sensing was realized for cotinine in the range of 100–1000 ng/mL (R2 = 0.96) in PBS medium. Undiluted urine samples from non-smokers exhibited an Ag-oxidation rate three times higher than the smoker’s urine samples. For 1:8 diluted urine samples (smokers), the proposed paper-based competitive immunochromatography coupled with an enzyme-modified electrode differentiated positive and negative samples and exhibited cotinine discrimination at levels higher than 12 ng/mL. This novel sensing platform can potentially be combined with a smartphone as a reader unit.

Antibiotics are classes of antimicrobial substances that are administered widely in the field of veterinary science to promote animal health and feed efficiency. Cattle-administered antibiotics hold a risk of passing active residues to milk, during the milking process. This becomes a public health concern as these residues can cause severe allergic reactions to sensitive groups and considerable economic losses to the farmer. Hence, to ensure that the produced milk is safe to consume and adheres to permissible limits, an on-farm quick and reliable test is essential. This study illustrates the design and development of a microfluidic paper biosensor as a proof-of-concept detection system for gentamicin in milk. Localized surface plasmon resonance (LSPR) properties of gold nanoparticles have been explored to provide the user a visual feedback on the test, which was also corroborated by RGB analysis performed using Image J. The assay involves the use of a short stretch of single stranded DNA, called aptamer, which is very specific to the gentamicin present in the milk sample. The camera-based LOD for the fabricated paper device for milk samples spiked with gentamicin was calculated to be 300 nM, with a reaction time of 2 min.

Paper-based microfluidic devices are an attractive platform for developing low-cost, point-of-care diagnostic tools. As paper-based devices&rsquo, detection chemistries become more complex, more complicated devices are required, often entailing the sequential delivery of different liquids or reagents to reaction zones. Most research into flow control has been focused on introducing delays. However, delaying the flow can be problematic due to increased evaporation leading to sample loss. We report the use of a CO2 laser to uniformly etch the surface of the paper to modify wicking speeds in paper-based microfluidic devices. This technique can produce both wicking speed increases of up to 1.1&times, faster and decreases of up to 0.9&times, slower. Wicking speeds can be further enhanced by etching both sides of the paper, resulting in wicking 1.3&times, faster than unetched channels. Channels with lengthwise laser-etched grooves were also compared to uniformly etched channels, with the most heavily grooved channels wicking 1.9&times, faster than the fastest double-sided etched channels. Furthermore, sealing both sides of the channel in packing tape results in the most heavily etched channels, single-sided, double-sided, and grooved, wicking over 13&times, faster than unetched channels. By selectively etching individual channels, different combinations of sequential fluid delivery can be obtained without altering any channel geometry. Laser etching is a simple process that can be integrated into the patterning of the device and requires no additional materials or chemicals, enabling greater flow control for paper-based microfluidic devices.

High concentrations of certain nutrients, including phosphate, are known to lead to undesired algal growth and low dissolved oxygen levels, creating deadly conditions for organisms in marine ecosystems. The rapid and robust detection of these nutrients using a colorimetric, paper-based system that can be applied on-site is of high interest to individuals monitoring marine environments and others affected by marine ecosystem health. Several techniques for detecting phosphate have been reported previously, yet these techniques often suffer from high detection limits, reagent instability, and the need of the user to handle toxic reagents. In order to develop improved phosphate detection methods, the commonly used molybdenum blue reagents were incorporated into a paper-based, colorimetric detection system. This system benefited from improved stabilization of the molybdenum blue reagent as well as minimal user contact with toxic reagents. The colorimetric readout from the paper-based devices was analyzed and quantified using RGB analyses (via ImageJ), and resulted in the detection of phosphate at detection limits between 1.3 and 2.8 ppm in various aqueous media, including real seawater.

Norovirus is one of the leading causes of gastroenteritis, acute vomiting, intense diarrhoea, acute pain in the stomach, high fever, headaches, and body pain. Conventional methods of detection gave us very promising results but had disadvantages such as low sensitivity, cost ineffectiveness, reduced specificity and selectivity, etc. Therefore, biosensors can be a viable alternative device which can overcome all setbacks associated with the conventional method. An electrochemical sensor based on oxidized graphitic carbon nitride (Ox-g-C3N4) modified electrochemical paper-based analytical device (ePAD) was fabricated for the detection of norovirus DNA. The synthesized Ox-g-C3N4 nanosheets were characterized by field emission scanning electron microscopy (FESEM), X-ray Diffraction (XRD), UV-Vis spectroscopy and X-Ray Photoelectron Spectroscopy. The capture probe DNA (PDNA) modified electrodes were characterized by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). These two characterization techniques were also employed to find the optimal scan rate, response time and temperature of the fabricated sensor. The fabricated biosensor showed a limit of detection (LOD) of 100 fM. Furthermore, the specificity of the reported biosensor was affirmed by testing the response of capture probe DNA with oxidized graphitic carbon nitride (PDNA/Ox-g-C3N4) modified ePAD on the introduction of a non-complimentary DNA. The fabricated ePAD sensor is easy to fabricate, cost effective and specific, and requires a minimum analysis time of 5 s.

Although the established ELISA-based sensing platforms have many benefits, the importance of cytokine and cancer biomarkers detection for point-of-care diagnostics has propelled the search for more specific, sensitive, simple, accessible, yet economical sensor. Paper-based biosensor holds promise for future in-situ applications and can provide rapid analysis and data without the need to conduct in a laboratory. Electrochemical detection plays a vital role in interpreting results obtained from qualitative assessment to quantitative determination. In this review, various factors affecting the design of an electrochemical paper-based biosensor are highlighted and discussed in depth. Different detection methods, along with the latest development in utilizing them in cytokine and cancer biomarkers detection, are reviewed. Lastly, the fabrication of portable electrochemical paper-based biosensor is ideal in deliberating positive societal implications in developing countries with limited resources and accessibility to healthcare services. Ministry of Education (MOE) Published version The authors would like to acknowledge the financial support for this work funded by Singapore Ministry of Education Academic Research Fund Tier 1 (MOE-2018-T1-001-004).

In this study, a highly efficient carbon-supported Pd catalyst for the direct ethanol fuel cell was developed by electrodepositing nanostructured Pd on oxygen plasma-treated carbon fiber paper (Pd/pCFP). The oxygen plasma treatment has been shown to effectively remove the surface organic contaminants and add oxygen species onto the CFP to facilitate the deposition of nano-structured Pd on the surface of carbon fibers. Under the optimized and controllable electrodeposition method, nanostructured Pd of ~10 nm can be easily and evenly deposited onto the CFP. The prepared Pd/pCFP electrode exhibited an extraordinarily high electrocatalytic activity towards ethanol oxidation, with a current density of 222.8 mA mg−1 Pd. Interestingly, the electrode also exhibited a high tolerance to poisoning species and long-term stability, with a high ratio of the forward anodic peak current density to the backward anodic peak current density. These results suggest that the Pd/pCFP catalyst may be a promising anodic material for the development of highly efficient direct alcohol fuel cells.

We report a paper-based electrochemical immunosensor made with sustainable materials to detect aflatoxin B1 (AFB1), a highly toxic, carcinogenic mycotoxin found in food. The immunosensor was prepared with a waterproof paper substrate and low-cost graphite-based conductive ink through a simple cut-printing method. The working electrode was functionalized with a drop-cast film of multiwalled carbon nanotubes (MWCNT)/chitosan on which a layer of anti-AFB1 monoclonal antibodies was immobilized covalently. The architecture of the immunosensor was confirmed with polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS) and electrochemical impedance spectroscopy (EIS), including the effective immobilization of the active layer of anti-AFB1. With EIS as the principle of detection, the immunosensor could detect AFB1 in the range from 1 to 30 ng&middot, mL&minus, 1, and detection limit of 0.62 ng&middot, 1. This sensitivity is sufficient to detect AFB1 in food according to regulatory agencies. The immunosensor exhibited good repeatability, reproducibility, stability, and selectivity in experiments with a possible interferent. Furthermore, detection of AFB1 in maize flour samples yielded recovery of 97&ndash, 99%, in a demonstration of the possible use of the paper-based immunosensor to detect AFB1 using extraction solutions from food samples.

Paper-based microfluidic devices have the potential of being a low-cost platform for diagnostic devices. Electrical circuit analogy (ECA) model has been used to model the wicking process in paper-based microfluidic devices. However, material characteristics such as absorption capacity cannot be included in the previous ECA models. This paper proposes a new model to describe the wicking process with liquid absorption in a paper strip. We observed that the fluid continues to flow in a paper strip, even after the fluid reservoir has been removed. This phenomenon is caused by the ability of the paper to store liquid in its matrix. The model presented in this paper is derived from the analogy to the current response of an electric circuit with a capacitance. All coefficients in the model are fitted with data of capillary rise experiments and compared with direct measurement of the absorption capacity. The theoretical data of the model agrees well with experimental data and the conventional Washburn model. Considering liquid absorption capacity as a capacitance helps to explain the relationship between material characteristics and the wicking mechanism.

There is a huge demand for rapid, reliable and low-cost methods for the analysis of heavy metals in drinking water, particularly in the range of sub-part per billion (ppb). In the present work, we describe the preparation, characterization and analytical performance of the disposable sensor to be employed in Square Wave Anodic Stripping Voltammetry (SWASV) for ultra-trace simultaneous determination of cadmium and lead. The electrode consists of graphene paper-perfluorosulfonic ionomer-bismuth nano-composite material. The electrode preparation implies a key step aimed to enhance the Bi3+ adsorption into nafion film, prior to the bismuth electro-deposition. Finely dispersed bismuth nanoparticles embedded in the ionomer film are obtained. The electrode was characterized by Scanning Electron Microscopy (SEM), Energy Dispersive X-ray Spectroscopy (EDX), Atomic Force Microscopy (AFM), X-ray Photoelectron Spectroscopy (XPS) and Electrochemical Impedance Spectroscopy (EIS). The electrode shows a linear response in the 5&ndash, 100 ppb range, a time-stability tested up to almost three months, and detection limits up to 0.1 ppb for both Pb2+ and Cd2+. The electrode preparation method is simple and low in cost and the obtained analytical performance is very competitive with the state of art for the SWASV determination of Pb2+ and Cd2+ in solution.

Microfluidic paper-based analytical devices (µPADs) have become promising tools offering various analytical applications for chemical and biological assays at the point-of-care (POC). Compared to traditional microfluidic devices, µPADs offer notable advantages; they are cost-effective, easily fabricated, disposable, and portable. Because of our better understanding and advanced engineering of µPADs, multistep assays, high detection sensitivity, and rapid result readout have become possible, and recently developed µPADs have gained extensive interest in parallel analyses to detect biomarkers of interest. In this review, we focus on recent developments in order to achieve µPADs with high-throughput capability. We discuss existing fabrication techniques and designs, and we introduce and discuss current detection methods and their applications to multiplexed detection assays in relation to clinical diagnosis, drug analysis and screening, environmental monitoring, and food and beverage quality control. A summary with future perspectives for µPADs is also presented.

We propose a flexible, binder-free and free-standing carbonaceous paper fabricated via electrostatic spray deposition using reduced graphene oxide/carbon nanotube (rGO/CNT) as a promising electrode material for flexible sodium-ion batteries (NIBs). The as-prepared rGO/CNT paper exhibits a three-dimensional (3D) layered structure by employing rGO as conductive frameworks to provide sodium-storage active sites and CNT as spacer to increase rGO interlayer distance and benefit the diffusion kinetics of sodium ions. Consequently, the rGO/CNT paper delivers an enhanced sodium ion storage capacity of 166.8 mAh g&minus, 1 at 50 mA g-1, retaining an average capacity of 101.4 mAh g&minus, 1 when current density sets back 100 mA g&minus, 1 after cycling at various current rates. An average capacity of 50 mAh g&minus, 1 at 200 mA g&minus, 1 was stabilized when cycling up to 300 cycles. The well-maintained electrochemical performance of free-standing rGO/CNT paper is due to the well-established hybrid 3D nanostructures, which demonstrates our carbon based material fabricated by a facile approach can be applied as one of the high-performance and low-cost electrode materials for applications in flexible energy storage devices.

This work describes a novel approach to produce high quality release paper at lower cost than traditional methods. The anti-adhesive properties of release paper require the use of expensive machine glazed kraft or &ldquo, Glassine&rdquo, paper as the paper base. A series of polymer coatings including polyvinyl alcohol, carboxymethyl cellulose, polyethylene vinyl acetate, and polystyrene were chemically synthesized and coated onto a low cost pulp paper base. Surface roughness (Sa) and smoothness coefficients (k) were determined by atomic force microscopy (AFM), and the interactions between the polymer coating and base paper were investigated by Raman spectroscopy. Studies show the use of polyethylene vinyl acetate (PEVA) as a pre-coating layer on low cost pulp paper exhibits similar anti-adhesive properties as higher cost paper bases. In low margin markets such as the production of release paper, decreases in cost are critical to industry survival.

By analyzing the three-dimensional digital model of a real carbon paper gas diffusion layer (GDL) reconstructed by X-ray computed tomography (CT), it was found that fibers are not distributed at any angle but within a certain range. The fiber orientation can be represented by fiber pitch (i.e., the angle between a single fiber and the in-plane direction). The effect of fiber orientation on stochastic reconstruction and transport properties (permeability) was investigated in this paper to find which fiber pitch range can achieve a better GDL on fluid flow. First, the actual fiber pitch was measured by analyzing SGL-24BA images obtained by X-ray CT. Also, seven different ranges of fiber pitch were randomly chosen to reconstruct GDL. Then, the permeability of these digital models was calculated using the Lattice Bolzmann Method (LBM) and discussed to obtain the fiber pitch range of the optimal permeability. The results show that the mean fiber pitch of SGL-24BA is 2.40&deg, and the individual values are all less than 6&deg, also, the permeability of the through-plane direction increases gradually as the range of fiber pitch increases, which can be used for the structural design of carbon paper GDL.

Paper has recently found widespread applications in biomedical fields, especially as an alternative scaffolding material for cell cultures, owing to properties such as its fibrous nature, porosity and flexibility. However, paper on its own is not an optimal material for cell cultures as it lacks adhesion moieties specific to mammalian cells, and modifications such as hydrogel integration and chemical vapor deposition are necessary to make it a favorable scaffolding material. The present study focuses on modification of filter paper through electrospin-coating and dip-coating with polycaprolactone (PCL), a promising biomaterial in tissue engineering. Morphological analysis, evaluation of cell viability, alkaline phosphatase (ALP) activity and live/dead assays were conducted to study the potential of the modified paper-based scaffold. The results were compared to filter paper (FP) and electrospun PCL (ES-PCL) as reference samples. The results indicate that electrospin-coating paper is a simple and efficient way of modifying FP. It not only improves the morphology of the deposited electrospun layer through reduction of the fiber diameter by nearly 75%, but also greatly reduces the scaffold fabrication time compared to ES-PCL. The biochemical assays (Resazurin and ALP) indicate that electrospin-coated filter paper (ES-PCL/FP) provides significantly higher readings compared to all other groups. The live/dead results also show improved cell-distribution and cell-scaffold attachment all over the ES-PCL/FP.

Traditionally, the acidity of paper-based relics was determined by an extraction method and using a pH meter. This method could not obtain the total acidity of the paper-based relics because it only detected the concentration of free protons in the aqueous soaking solution. To overcome this defect, a new method for determining the total acidity of paper-based relics has been established by using quaternary alloy quantum dots. The quantum dots, CdZnSeS, modified by p-Aminothiophenol (pATP) were prepared, and their composition and structure were characterized. The fluorescence behavior of prepared quantum dots with acidity was investigated. The following results were obtained. The fluorescence of CdZnSeS-pATP quantum dots could decrease with increases in acidity because pATP dissociated from the surfaces of the quantum dots due to protons or undissociated weak acids. Based on this feature, a method for determining the acidity of paper-based relics was constructed, and this method was used to evaluate the acidity of actual paper-based relics. Obviously, for a given paper sample, since both free protons and bound protons can be determined by this method, the acidity measured by this method is more reasonable than that by pH meter.

Maize stem sculptures, produced during the 16th and 17th centuries in New Spain (today, Mexico) are a clear example of the convergence of the artistic traditions from the American indigenous populations and European influence. This typology of sculptures is not limited to the Americas, as the examples found in European countries have shown. Therefore, a detailed technological investigation is required to correctly classify them. This work presents the interdisciplinary and multianalytical investigation of a 16th-century sculpture made with a maize stem preserved in Guadalajara city, Mexico. We used a set of techniques, such as CT, SEM-EDX, μ-FTIR, and μ-Raman, to study, from a macro to a micro level, the structure, the polychromy, and the modification of the sculpture. The results showed the use of maize stems, paper, and wood in the construction of the sculpture and the use of the traditional polychromy, as well as the numerous modifications that changed its appearance considerably resulting in its misclassification. We were able to associate the statue with the Cortés workshop (Mexico City region), probably produced in the decade of 1580, and track its liturgical use and historical development through the centuries.

Enzyme immobilization on different supports has emerged as an efficient and cost-effective tool to improve their stability and reuse capacity. This work aimed to produce a stable immobilized multienzymatic system of xylanase and filter paper-ase (FPase) onto magnetic chitosan using genipin as a cross-linking agent and to evaluate its biochemical properties and reuse capacity. A mixture of chitosan magnetic nanoparticles, xylanase, and FPase was covalently bonded using genipin. Immobilization yield and efficiency were quantified. The activity of free and immobilized enzymes was quantified at different values of pH, temperature, substrate concentration (Km and Vmax), and reuse cycles. The immobilization yield, immobilization efficiency, and activity recovery were 145.3% ± 3.06%, 14.8% ± 0.81%, and 21.5% ± 0.72%, respectively, measured as the total hydrolytic activity. Immobilization confers resistance to acidic/basic conditions and thermal stability compared to the free form. Immobilization improved 3.5-fold and 78-fold the catalytic efficiency (Kcat/Km) of the xylanase and filter paper-ase activities, while immobilized xylanase and FPase could be reused for 34 min and 43 min, respectively. Cross-linking significantly improved the biochemical properties of immobilized enzymes, combined with their simplicity of reuse due to the paramagnetic property of the support. Multienzyme immobilization technology is an important issue for industrial applications.

Microparticles encapsulated with orange essential oil were prepared by improved emulsifying solvent volatilization technology, and modified with chitosan to improve their loading and adhesion properties on paper. Characterization was performed by Zetasizer Nano ZS instrument, transmission electron microscope (TEM), scanning electron microscopy (SEM), Fourier transform infrared spectrometer (FTIR) spectroscopy, thermogravimetric analyzer (TGA), gas-chromatography-mass spectrometry (GC-MS) and the ultrafast GC Electronic Nose Heracles II, etc. The results showed that for poly (lactic-co-glycolic acid) (PLGA) microparticles and chitosan&ndash, PLGA microparticles, respectively, the particle sizes were 233.2 and 277.6 nm, loading capacity was 19.17% and 24.36%, Zeta potential was &minus, 8.27 and 5.44 mV, adhesive capacity was 76.32 and 324.84 mg/g, and encapsulation efficiency was 93.23% and 94.06%. GC-MS demonstrated that the embedding process minimally effected the aroma quality of orange essential oil. The ultrafast GC Electronic Nose Heracles II showed that chitosan&ndash, PLGA microparticles could effectively slow the release of the orange essential oil. Therefore, this work provides a proposal for a better understanding of biodegradable functional packaging paper.

Humidity sensors are indispensable for various electronic systems and instrumentations. To develop a new humidity sensing mechanism is the key for the next generation of sensor technology. In this work, a novel flexible paper-based current humidity sensor is proposed. The developed alternating current electroluminescent devices (ACEL) consist of the electroless plating Ni on filter paper and silver nanowires (AgNWs) as the bottom and upper electrodes, and ZnS:Cu as the phosphor layer, respectively. The proposed humidity sensor is based on ACEL with the paper substrate and the ZnS:Cu phosphor layer as the humidity sensing element. The moisture effect on the optical properties of ACELs has been studied firstly. Then, the processing parameters of the paper-based ACELs such as electroless plated bottom electrode and spin-coated phosphor layer as a function of the humidity-sensitive characteristics are investigated. The sensing mechanism of the proposed sensor has been elucidated based on the Q ~ V analysis. The sensor exhibits an excellent linearity ( R 2 = 0.99965 ) within the humidity range from 20% to 90% relative humidity (RH) and shows excellent flexibility. We also demonstrate its potential application in postharvest preservation where the EL light is used for preservation and the humidity can be monitored simultaneously through the current.

The following article summarizes United States Patent Application No. US20180052155A1, titled ‘Assay Devices and Methods’ (filed 16 August 2016, published 22 February 2018). While lateral flow assays (LFAs) have revolutionized point-of-care diagnostics by enabling accurate, inexpensive, and rapid detection of biomarkers, they typically do not provide quantitative results. Hence, there is a significant need for quantitative assays at the point of care. This patent summary describes a novel method of chronometric biomarker quantitation via enzymatic degradation of a metastable gelatin-based biomatrix, principally suited for use in paper-based microfluidic devices (microPADs). This new quantitation mechanism was designed to meet the ASSURED criteria for point-of-care diagnostic devices laid forth by the World Health Organization and may ultimately provide increased access to healthcare, at a significantly reduced cost, around the world.

This work describes the use of a paper substrate for electro-optical detection of toxic hydrogen sulfide (H2S) gas. For electrical detection, a chemiresistive type of gas sensor was developed. Ultrathin gold film electrodes (UTGFE) were produced by physical vapor deposition of gold on nanostructured latex-coated paper substrate. The gas-sensing film was deposited on the electrodes by inkjet printing. The sensing films were characterized by atomic force microscopy, X-ray photoelectron spectroscopy and conductometry. The sensing films showed more than seven orders of magnitude change in resistance when exposed to as low as 1 part per million (ppm) H2S gas at room temperature. Besides resistive response, the change in color of the sensing films was studied on a paper substrate, both as a function of print density of the sensing material and H2S concentration. For quantification of the analyte the red, green and blue color deconvolution was performed on the pictures of the paper strip indicator using an open source software. A clear response was obtained from the blue channel. The inexpensive disposable color strips produced on the paper substrate can be used for qualitative and quantitative detection (as low as 1.5 ppm) of H2S gas. Electro-Optical Gas Sensor Consisting of Nanostructured Paper Coating and an Ultrathin Sensing Element

Paper-based relics, which are an important part of cultural heritage worldwide, are at risk of imminent damage from various environmental sources. To protect them, the atmospheric pressure plasma polymerization of hexamethyldisiloxane (HMDSO) precursor has been explored on paper-based relics in situ. The macro and micro images taken during this process suggest that the in situ plasma treatment does not change the macro morphology and the micro structure of the treated paper-based relic samples. On the other hand, plasma treatment causes the polymerization of the HMDSO which then produces nanoparticles deposited onto the paper-based relics. These nanoparticles provide good waterproof properties with large static water contact angles and smaller rolling angles, which protect the paper-based relics from water penetration. Moreover, since the nanoparticles are deposited onto the fibers, waterproof fastness is ensured. Also, the examined mechanical properties of the treated and untreated paper-based relics indicate that the atmospheric pressure plasma treatment does not affect the strength of the paper very much. The results in this study show that atmospheric pressure plasma treatment with the use of HMDSO precursor is a good method to preserve paper-based relics.

In this paper, we report the fabrication of highly conductive poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT:PSS)/cellulose nanofiber (CNF) nanocomposite paper with excellent flexibility through post-treatment with an organic solvent. The post-treated PEDOT:PSS/CNF porous nanocomposite papers showed a lower sulfur content, indicating the removal of residual PSS. The electrical conductivity of PEDOT:PSS/CNF porous nanocomposite paper was increased from 1.05 S/cm to 123.37 S/cm and 106.6 S/cm by post-treatment with dimethyl sulfoxide (DMSO) and ethylene glycol (EG), respectively. These values are outstanding in the development of electrically conductive CNF composites. Additionally, the highly conductive nanocomposite papers showed excellent bending stability during bending tests. Cyclic voltammetry (CV) showed a Faradaic redox reaction and non-Faradaic capacitance due to the redox activity of PEDOT:PSS and large surface area, respectively. Electrochemical energy storage ability was evaluated and results showed that capacitance improved after post-treatment. We believe that the highly conductive PEDOT:PSS/CNF porous nanocomposite papers with excellent flexibility described here are potential candidates for application in porous paper electrodes, flexible energy storage devices, and bioengineering sensors.

Food safety and access to systematic approaches for ensuring detection of food hazards is an important issue in most developing countries. With the arrival of paper-based analytical devices (&micro, PADs) as a promising, rapid, easy-to-use, and low-cost analytical tool, we demonstrated a simple microfluidic-based titration study for the analysis of packaged fruit juices. Similar, to the titration experiments using traditional glassware in chemistry laboratories, in this study the titration experiments were developed using paper microfluidics for the analysis of several analytes such as pH, vitamin C, sugars, and preservatives present in the packaged fruit juices. The allergen found commonly in dairy based mixtures and the non-pathogenic biochemical component responsible for food spoilage in cider based fruit juices were also determined. The results obtained using paper microfluidics were compared with those obtained using a conventional spectrophotometric technique. Finally, a paper microfluidics based multiplexed sensor was developed for the analysis of common nutritional ingredients, an allergen, and a non-pathogenic byproduct present in packaged fruit juices on a single platform. Overall, the results presented in this study reveal that the proposed paper microfluidic assisted colorimetric multiplexed sensor offers a quick and reliable tool for on-spot routine analysis for food safety applications.

Here, we aimed to boost antibacterial performance of cellulose fibers for paper sheet application. Therefore, TiO2 nanoparticles have been used with controlled loading onto the surface of the fibers. A simple and facile composite preparation route based on ultrasound and mechanical assisted stirring has been developed. We tested cellulose paper enriched by TiO2 from 1.0 wt% to 8.0 wt%, respectively. Antibacterial performance has been studied against Staphylococcus aureus and Escherichia coli bacteria. Studies showed that all composites exhibit significant capability to reduce living cells of S. aureus and E. coli bacteria at least 60%. The simplicity, low cost, and reproducibility of the prepared method indicates the potential to be scaled up for industrial applications.

Food safety issues caused by pesticide residue have exerted far-reaching impacts on human daily life, yet the available detection methods normally focus on surface residue rather than pesticide penetration to the internal area of foods. Herein, we demonstrated gold nanoparticle (AuNP)-immersed paper imprinting mass spectrometry imaging (MSI) for monitoring pesticide migration behaviors in various fruits and vegetables (i.e., apple, cucumber, pepper, plum, carrot, and strawberry). By manually stamping food tissues onto AuNP-immersed paper, this method affords the spatiotemporal visualization of insecticides and fungicides within fruits and vegetables, avoiding tedious and time-consuming sample preparation. Using the established MSI platform, we can track the migration of insecticides and fungicides into the inner region of foods. The results revealed that both the octanol-water partition coefficient of pesticides and water content of garden stuffs could influence the discrepancy in the migration speed of pesticides into food kernels. Taken together, this nanopaper imprinting MSI is poised to be a powerful tool because of its simplicity, rapidity, and easy operation, offering the potential to facilitate further applications in food analysis. Moreover, new perspectives are given to provide guidelines for the rational design of novel pesticide candidates, reducing the risk of food safety issues caused by pesticide residue.

Microfluidic paper-based analytical devices (microPADs) have emerged as a promising platform for point-of-care diagnostic devices. While the inherent wicking properties of microPADs allow for fluid flow without supporting equipment, this also presents a major challenge in achieving robust fluid control, which becomes especially important when performing complex multi-step assays. Herein, we describe an ideal method of fluid control mediated by wax-printed fluidic time delays. This method relies on a simple fabrication technique, does not utilize chemicals/reagents that could affect downstream assays, is readily scalable, and has a wide temporal range of tunable fluid control. The delays are wax printed on both the top and bottom of pre-fabricated microPAD channels, without subsequent heating, to create hemi-/fully-enclosed channels. With these wax printed delays, we were able to tune the time it took aqueous solutions to wick across a 25 mm-long channel between 3.6 min and 13.4 min. We then employed these fluid delays in the sequential delivery of four dyes to a test zone. Additionally, we demonstrated the automation of two simple enzymatic assays with this fluid control modality. This method of fluid control may allow future researchers to automate more complex assays, thereby further advancing microPADs toward real-world applications.

Invasive alien plant species (IAPS) are one of the biggest challenges in European ecosystems, displacing local vegetation, destroying agricultural land, and causing billions of dollars of damage to the European economy every year. Many of them are removed daily and mainly burned. In this work, we investigated the possibilities of using plants as feedstock for paper production. Papers made from three invasive alien plants, i.e., Knotweed, Goldenrod, and Black locust, were studied and compared with commercial office paper. The study included testing of: (1) structural properties—basic physical properties, grammage, thickness, density and specific volume, moisture content, and ash content, (2) physical and dynamic mechanical properties—tensile strength, Clark stiffness, viscoelastic properties, (3) colorimetric properties of prints, (4) effect of UV light on ageing, and (5) study of cellulose fiber structure and morphology by microscopy. The results suggested that the paper produced can be used as commercial office paper, considering that the paper is slightly dyed. Such papers can also be used for special purposes that present a natural style and connection to nature. The papers produced can also be used for printing documents that are meant to be kept.

Paper spray ionization mass spectrometry (PSI-MS) is a direct MS analysis technique with several reported bacterial metabolomics applications. As with most MS-based bacterial studies, all currently reported PSI-MS bacterial analyses have focused on the chemical signatures of the cellular unit. One dimension of the bacterial metabolome that is often lost in such analyses is the exometabolome (extracellular metabolome), including secreted metabolites, lipids, and peptides. A key component of the bacterial exometabolome that is gaining increased attention in the microbiology and biomedical communities is extracellular vesicles (EVs). These excreted structures, produced by cells in all domains of life, contain a variety of biomolecules responsible for a wide array of cellular functions, thus representing a core component of the bacterial secreted metabolome. Although previously examined using other MS approaches, no reports currently exist for a PSI-MS analysis of bacterial EVs, nor EVs from any other organism (exosomes, ectosomes, etc.). PSI-MS holds unique analytical strengths over other commonly used MS platforms and could thus provide an advantageous approach to EV metabolomics. To address this, we report a novel application representing, to our knowledge, the first PSI-MS analysis of EVs from any organism (using the human gut resident Oxalobacter formigenes as the experimental model, a bacterium whose EVs were never previously investigated). In this report, we show how we isolated and purified EVs from bacterial culture supernatant by EV-specific affinity chromatography, confirmed and characterized these vesicles by nanoparticle tracking analysis, analyzed the EV isolate by PSI-MS, and identified a panel of EV-derived metabolites, lipids, and peptides. This work serves as a pioneering study in the field of MS-based EV analysis and provides a new, rapid, sensitive, and economical approach to EV metabolomics.

Development of paper-based sensors that do not suffer with humidity interference is desirable for practical environmental applications. In this work, a laser processing method was reported to effectively modulate the cross-sensitivity to humidity of ZnO-based UV (Ultraviolet) sensors printed on paper substrate. The results reveal that the laser induced zinc oxide (ZnO) surface morphology contributes to the super-hydrophobicity of the printed ZnO nanoparticles, reducing humidity interference while enhancing UV sensitivity. Herein, this conducted research highlights for the first time that laser processing is an attractive choice that reduces the cross-sensitivity to water vapor in the UV sensing response of ZnO-based devices printed on paper, paving the way to low-cost and sophisticated paper-based sensors.

Paraquat is a highly toxic herbicide. Paraquat poisoning is often fatal and is an important public health threat in many places. The quick identification and timely initiation of treatment based on timely analysis of the paraquat concentration in urine/serum could improve the prognosis for patients. However, current paraquat concentration measurements are time-consuming and difficult to implement due to the expensive and bulky equipment required. To address these practical challenges, paper-based devices have emerged as alternative diagnostic tools for improving point-of-care testing. In this study, we demonstrate the successful use of a paper-based analytical device for the accurate detection of urine paraquat concentration. The developed paper-based analytical device employs colorimetric paraquat concentration measurements. The R2 value for the urine paraquat standard curve was 0.9989, with a dynamic range of 0&ndash, 100 ppm. The limit of detection was 3.01 ppm. Two other optical-based approaches, Spectrochip and NanoDrop, were used for comparison. The results suggest that the developed paper-based analytical device is comparable to other colorimetric measurements, as determined by Bland&ndash, Altman analysis. The device was clinically validated using urine from six paraquat-poisoned patients. The results prove that the developed paper-based analytical device is accurate, easy-to-use, and efficient for urine paraquat concentration measurement, and may enable physicians to improve clinical management.

To transform from reactive to proactive healthcare, there is an increasing need for low-cost and portable assays to continuously perform health measurements. The paper-based analytical devices could be a potential fit for this need. To miniaturize the multiplex paper-based microfluidic analytical devices and minimize reagent use, a fabrication method with high resolution along with low fabrication cost should be developed. Here, we present an approach that uses a desktop pen plotter and a high-resolution technical pen for plotting high-resolution patterns to fabricate miniaturized paper-based microfluidic devices with hundreds of detection zones to conduct different assays. In order to create a functional multiplex paper-based analytical device, the hydrophobic solution is patterned on the cellulose paper and the reagents are deposited in the patterned detection zones using the technical pens. We demonstrated the effect of paper substrate thickness on the resolution of patterns by investigating the resolution of patterns on a chromatography paper with altered effective thickness. As the characteristics of the cellulose paper substrate such as thickness, resolution, and homogeneity of pore structure affect the obtained patterning resolution, we used regenerated cellulose paper to fabricate detection zones with a diameter as small as 0.8 mm. Moreover, in order to fabricate a miniaturized multiplex paper-based device, we optimized packing of the detection zones. We also showed the capability of the presented method for fabrication of 3D paper-based microfluidic devices with hundreds of detection zones for conducting colorimetric assays., Scientific and Technological Research Council of Turkey (TÜBİTAK); 2232 International Fellowship for Outstanding Researchers Program