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In this transformative study, machine learning (ML) and t‐distributed stochastic neighbor embedding (t‐SNE) are employed to interpret intricate patterns in colorimetric images of cold atmospheric plasma (CAP)‐treated water. The focus is on CAP's therapeutic potential, particularly its ability to generate reactive oxygen and nitrogen species (RONS) that play a crucial role in antimicrobial activity. RGB, HSV, LAB, YCrCb, and grayscale color spaces are extracted from the colorimetric expression of oxidative stress induced by RONS, and these features are used for unsupervised ML, employing density‐based spatial clustering of applications with noise (DBSCAN). The DBSCAN model's performance is evaluated using homogeneity, completeness, and adjusted rand index with a predictive data distribution graph. The best results are achieved with 3,3′,5,5′‐tetramethylbenzidine–potassium iodide colorimetric assay solution immediately after plasma treatment, with values of 0.894, 0.996, and 0.826. t‐SNE is further conducted for the best‐case scenario to evaluate the clustering efficacy and find the best combination of features to better present the results. Correspondingly, t‐SNE enhances clustering efficacy and adeptly handles challenging points. The approach pioneers dynamic and comprehensive solutions, showcasing ML's precision and t‐SNE's transformative visualization. Through this innovative fusion, complex relationships are unraveled, marking a paradigm shift in biomedical analytical methodologies. [ABSTRACT FROM AUTHOR]

In this transformative study, machine learning (ML) and t‐distributed stochastic neighbor embedding (t‐SNE) are employed to interpret intricate patterns in colorimetric images of cold atmospheric plasma (CAP)‐treated water. The focus is on CAP's therapeutic potential, particularly its ability to generate reactive oxygen and nitrogen species (RONS) that play a crucial role in antimicrobial activity. RGB, HSV, LAB, YCrCb, and grayscale color spaces are extracted from the colorimetric expression of oxidative stress induced by RONS, and these features are used for unsupervised ML, employing density‐based spatial clustering of applications with noise (DBSCAN). The DBSCAN model's performance is evaluated using homogeneity, completeness, and adjusted rand index with a predictive data distribution graph. The best results are achieved with 3,3′,5,5′‐tetramethylbenzidine–potassium iodide colorimetric assay solution immediately after plasma treatment, with values of 0.894, 0.996, and 0.826. t‐SNE is further conducted for the best‐case scenario to evaluate the clustering efficacy and find the best combination of features to better present the results. Correspondingly, t‐SNE enhances clustering efficacy and adeptly handles challenging points. The approach pioneers dynamic and comprehensive solutions, showcasing ML's precision and t‐SNE's transformative visualization. Through this innovative fusion, complex relationships are unraveled, marking a paradigm shift in biomedical analytical methodologies.

Soil nutrition is a key pillar in agricultural productivity. However, point-of-need testing for soil nutrition is not readily available in resource-limited settings such as Kenya. We set out to study the perceived need for soil testing among farmers in this country. A group of 547 farmers from Murang'a and Kiambu counties in central Kenya were recruited through multi-stage sampling to help assess the perceptions and willingness to pay (WTP) toward a prototype technology for surveillance of in-situ soil nutrition. The technology is based on a cafetière-style filter system for extraction and a microfluidic paper-based analytical device (μPAD) for nutrient readout. We employed the double bounded choice contingent valuation method (CVM) to analyze the willingness of farmers to accept and pay for the prototype if the technology was available on the market. It was found that currently, only 1.5 % of farmers carry out soil testing. The high costs of analysis at testing centers, which are often far from the farmers, are among the main reasons contributing to the majority of farmers not testing their soils. The farmers surveyed were generally willing to make their soil data publicly accessible, especially to extension officers. CVM showed that uncontrolled WTP had a 94.24 % premium above KSh1,000 ($6.60) incurred by using the existing rapid testing method. Factoring the control variables and disaggregating the model into gender categories, the findings showed that youth, women, and men had WTP values of KSh1,612.53 ($10.75), KSh1,558.68 ($10.39), and KSh1,504.83 ($10.03), respectively, indicating that farmers can indeed pay for the convenience to test their soils in situ. Through the democratization of soil nutrition data, extension agents can enhance the improvement of agricultural productivity, which implies that farmers can commercialize their agricultural activities.

Microfluidic paper-based analytical devices (µPADs) have been successfully developed using dual detection: electrochemical and colorimetric systems. The µPADs have the potential to be used as Pb(II), Cd(II), and Cu(II) sensors to test the quality of water. The fabrication process uses hot embossing and screen-printing methods. The working electrode in the electrochemical zone was enhanced by the bismuth metal deposition process, while in the colorimetric zone, the gold nanoparticles modified with thioctic acid and dansylhydrazine (TA-Au-DNS) were used as a colorimetric sensor to detect Cu. The basic material of µPADs was characterized using a Fourier-transform infrared (FTIR) and a contact angle meter (CAM). In the electrochemical zone, the signals of square wave anodic stripping voltammetry (SWASV) resulted in good detection of Pb(II) and Cd(II) (from 0 to 100 ppb) with a limit of detection of 1.588 and 1.42 ppb, respectively. In the colorimetric zone, the performance of TA-Au-DNS for detecting Cu metal was obtained from readings through the red-green-blue (RGB) sensor as a miniature of µPADs reader. The LOD, LOQ, and average Vx0 (linearity values) in the detection of Cu(II) (from 58 to 100 ppb) are 8.51 ppb, 28.36 ppb, and 0.41%, respectively.

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Chlorpyrifos (CHL), profenofos (PRO) and cypermethrin (CYP) are widely used in combination to increase crop yields. However, these three pesticides can cause serious harm to human health and do not easily degrade. In this study, a novel visible paper sensor has been prepared successfully and different colorimetric reactions were utilized to detect the three pesticides simultaneously. The sensor was constructed by grafting a zwitterionic polymer onto a cellulose filter (CF) and placing it on a glass surface modified with PDMS. The branch shape was designed to form multiple detection areas, which were modified with specific pesticides and corresponding chromogenic reagents. The as-prepared colorimetric platform exhibited high sensitivity, a short detection time, a good linear response and a low detection limit (LOD) for the three pesticides (chlorpyrifos: y = 46.801 − 1.939x, R2 = 0.983, LOD = 0.235 mg/L; profenofos: y = 40.068 + 42.5x, R2 = 0.988, LOD = 4.891 mg/L; cypermethrin: y = 51.993 + 1.474x, R2 = 0.993, LOD = 4.053 mg/L). The comparison of the results obtained by the proposed paper sensor and those obtained by spectrophotometry further revealed the stability and reliability of the paper sensor. In particular, the color intensity of the interaction between the pesticides and coloring agents could be directly observed by the human eye. The consistency of the colorimetric/optical assay was proven in real target pesticide samples. Thus, this sensing strategy provides a portable, cost-effective, accurate and visualized paper platform, which could be suitable for application in the fruit and vegetable industry for monitoring CHL, PRO and CYP in parallel. [ABSTRACT FROM AUTHOR]

Nowadays, food safety has become a major concern for the sustainability of global public health. Through the production and distribution steps, food can be contaminated by either chemical hazards or pathogens, and the determination of these plays a critical role in the processes of ensuring food safety. Therefore, the development of analytical tools that can provide rapid screening of these hazards is highly necessary. Microfluidic paper-based analytical devices (µPADs) have advanced significantly in recent years as they are rapid and low-cost analytical screening tools for testing contaminated food products. This review focuses on recent developments of µPADs for various applications in the food safety field. A description of the fabrication of selected papers is briefly discussed, and evaluation of the μPADs' performance with regard to their precision and accuracy as well as their limits of detection is critically assessed. The advantages and disadvantages of these devices are highlighted. [ABSTRACT FROM AUTHOR]

The aim of this study is to employ a response surface methodology (RSM) to optimize experimental parameters in microfluidic paper-based analytical devices (µ-PADs). The independent parameters include concentrations and volumes of acetylcholinesterase, acetylthiocholine iodide, and 5,5-dithiobis-(2-nitrobenzoic acid). Their effect on mean color intensity was tested and optimized. From the analysis of variance, high regression and fitting values were obtained between the experimental and RSM predicted mean color intensity. Under optimum conditions, satisfactory linearity (R2 > 0.9990) in the range of 0.25–16 mg/L was obtained for the studied pesticides. The limit of detection (LOD) varies from 0.13 to 0.27 mg/L; high precision (RSD of 3.8–8.0%), reproducibility (RSD of 7.2–11.0%), and recovery (78–97%) were achieved. The RSM approach has been demonstrated to be more efficient than the traditional approach. It resulted in a µ-PAD system with less amount of reagent usage and better LOD compared with a univariate approach. [ABSTRACT FROM AUTHOR]

Accurately predicting durian maturity is a critically unresolved worldwide issue. Farmers currently determine durian ripeness based on their own observation and experience leading to inconsistencies in harvest timing. This reliance on human judgment often results in premature or overripe harvests, impacting fruit quality, yield, and market value. Existing technological solutions, such as sensors are often complex and require specialized expertise, hindering their adoption by farmers and consumers. Developing sensors that can accurately measure durian ripeness without damaging the fruit, are easy to use, and affordable remains a challenge. We introduce a microfluidic paper-based analytical device (μPAD) for on-site, safe matching to meet the demands of durian maturity evaluation. The μPAD automatically collected peduncle fluid without destroying the durian fruit for dual detection of total sugar and amino acid. For determining total sugar including sucrose, glucose, and fructose, several enzymatic steps were reduced to a single step of invertase for sucrose hydrolysis before total reducing sugar was measured using gold nanoparticle (AuNP) generation. Kinetics study of invertase on the μPAD showed V max and K m values of 1.42 mM min−1 and 2.17 mM, respectively, that agreed with the direct study of sucrose conversion. To increase device reliability, amino acid was also simultaneously measured with sugar using the simple ninhydrin test with the addition of SnCl 2. The developed sensor provided LODs of 3.50, 3.10, 3.30 μM, and 0.02 mg mL−1 for glucose, fructose, sucrose, and amino acid respectively. The μPADs were able to nondestructively discriminate between the mature and immature durians, showing high linear correlation with the standard dry weight method. The development of this μPAD technology has the potential to revolutionize durian cultivation practices, reduce post-harvest losses, and enhance the overall sustainability and profitability of the durian value chain, and can be further developed for maturity tests of other fruits. [Display omitted] • A user-friendly microfluidic paper-based device (μPAD) was introduced for on-site durian maturity evaluation. • The μPAD analyzed durian peduncle fluid, eliminating the need to damage the fruit itself. • The μPAD offered a single-step enzyme process to measure sugars and amino acids, simplifying traditional methods. • Kinetic study of invertase on paper substrate corresponded to the direct conversion of sucrose. • The μPAD accurately distinguished mature and immature durians, providing a reliable and affordable tool. [ABSTRACT FROM AUTHOR]

In this work, we developed an alternative manufacturing paper-based microfluidics method through 3D printing and wax filament. Microfluidic paper-based analytical devices (µPADs) are low-cost and easy-to-manufacture tools used for various chemical and biological analyses and studies. Paper-based microfluidics with wax has been limited as the manufacturers have discontinued most wax printing equipment. We aim to develop a low-cost and accessible manufacturing method that can replace conventional wax-on paper-based microfluidic manufacturing methods. Using highly available commercial 3D printing technology and wax filament, we could create hydrophobic wax barriers on the surface of different paper types. The properties and limits of this manufacturing method were characterized. Moreover, using this paper-based microfluidic manufacturing method, we were able to measure dopamine electrochemically using µPAD as a passive flow-based method in concentrations as low as 1 nM using injections as small as 15 µL.

A user-friendly, low-cost detector able to read the RGB indexes of microfluidic paper-based analytical devices (µPADs) was developed. The RGB-detector was built with 3D printing using PLA+ and reused Li-ion batteries. It is Arduino-based, which provides an easy interface between the sensor TCS3200, which reads the quadratic wave of the times corresponding to the RGB numbers, the Arduino itself, whose software translates the times into RGB values, and the touchscreen display, NX3224T028, which shows the results. This detector permits multi-sample analysis since it has a sample holder that can keep up to six µPADs simultaneously and rotate after the display's request. This work shows how the readings of the RGB indexes by the proposed RGB-detector implement the measurements' reproducibility. As a proof-of-concept, the RGB-detector application to a green array of µPADs for pH measurement coupled with chemometric analysis allowed us to achieve good results in terms of precision and agreement with the pH values measured by a classical pH-meter. [ABSTRACT FROM AUTHOR]

Tea (Camellia sinensis (L.) Kuntze—surname of German origin) is a popular beverage consumed worldwide due to its health benefits. Its quality depends on measuring features that may discriminate teas from distinct provenances. Protected designation of origin (PDO) is therefore a very useful label for tea quality evaluation. In the present work, antioxidant activity profiles obtained from microfluidic paper-based analytical devices (µPADs) were analyzed by chemometrics to determine the tea geographic origin. Based on the existing literature, we constructed a database containing chemical data from 26 samples and evaluated it by principal component analysis (PCA) coupled to linear discriminant analysis (LDA). Antioxidant activity was an effective LDA predictor for sample discrimination accomplishing accuracies from 75 to 82%. Modeling performance was favored by an external validation method. The best classification model was found using the first nine PCs as input variables. Training samples achieved a perfect success rate, while the test ones were predicted with 83% specificity, 100% sensitivity, and 90% overall accuracy. The modeling robustness was verified by integrating AUC (0.943) from ROC curve. The PCA-LDA approach taken here demonstrated that the teas coming from different countries can be correctly authenticated through µPADs, thus contributing to certificate samples PDO. [ABSTRACT FROM AUTHOR]

Soil nutrition is a key pillar in agricultural productivity. However, point-of-need testing for soil nutrition is not readily available in resource-limited settings such as Kenya. We set out to study the perceived need for soil testing among farmers in this country. A group of 547 farmers from Murang'a and Kiambu counties in central Kenya were recruited through multi-stage sampling to help assess the perceptions and willingness to pay (WTP) toward a prototype technology for surveillance of in-situ soil nutrition. The technology is based on a cafetière-style filter system for extraction and a microfluidic paper-based analytical device (μPAD) for nutrient readout. We employed the double bounded choice contingent valuation method (CVM) to analyze the willingness of farmers to accept and pay for the prototype if the technology was available on the market. It was found that currently, only 1.5 % of farmers carry out soil testing. The high costs of analysis at testing centers, which are often far from the farmers, are among the main reasons contributing to the majority of farmers not testing their soils. The farmers surveyed were generally willing to make their soil data publicly accessible, especially to extension officers. CVM showed that uncontrolled WTP had a 94.24 % premium above KSh1,000 ($6.60) incurred by using the existing rapid testing method. Factoring the control variables and disaggregating the model into gender categories, the findings showed that youth, women, and men had WTP values of KSh1,612.53 ($10.75), KSh1,558.68 ($10.39), and KSh1,504.83 ($10.03), respectively, indicating that farmers can indeed pay for the convenience to test their soils in situ . Through the democratization of soil nutrition data, extension agents can enhance the improvement of agricultural productivity, which implies that farmers can commercialize their agricultural activities., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

For the first time, in-tube gel electro-membrane microextraction (IT-G-EME) system followed by microfluidic paper-based analytical devices (µPADs) as a low-cost reading platform was fabricated for the speciation of trivalent chromium (Cr(III)) and hexavalent chromium (Cr(VI)) as model cationic and anionic compounds. In this new miniaturized extraction mode, a transparent narrow-bore polymeric tube was used as housing of the aqueous acceptor phase (AP, 30 µL), while an agarose gel membrane was placed as a micro plug (2.5 mm) at the beginning of the tube. A circular shape vial (1.5 mL, pH 5.5) containing chromium species as donor phase (DP) was connected between two tubes which previously filled with gel membrane and aqueous AP. By applying electric potential, the positively charged Cr(III) and negatively charged Cr(VI) in the DP migrated selectively toward cathodic and anodic tubes, respectively. After extraction, each AP was analyzed by µPAD, which had already been modified with diphenylcarbazide (DPC). Under the optimized extraction conditions, a good limit of detection (LOD) equal to 7.0 µg L–1 was achieved for both analytes, while the extraction recoveries for Cr(VI) and Cr(III) were 72% and 84%. In addition, the developed approach was used for the quantification of chromium species in water samples.

The global prevalence of diabetes mellitus, a serious chronic disease with fatal consequences for millions annually, is of utmost concern. The development of efficient and simple devices for monitoring glucose levels is of utmost significance in managing diabetes. The advancement of nanotechnology has resulted in the indispensable utilization of advanced nanomaterials in high-performance glucose sensors. Modulating the morphology and intricate composition of transition metals represents a viable approach to exploit their structure/function correlation, thereby achieving optimal electrocatalytic performance of the synthesized catalysts. Herein, a sensitive and rapid Cu-encapsulated Cu 2 S@nitrogen-doped carbon (Cu@Cu 2 S@N–C) hollow nanocubes-functionalized microfluidic paper-based analytical device (μ-PAD) was fabricated. Through a delicate sacrificial template/interface technique and thermal decomposition, inter-connected hollow networks were formed to boost the active sites, and the carbon shell was coated to protect Cu from being oxidation. For application, the constructed μ-PAD is used for glucose sensing utilizing an origami automated sample pretreatment system enabled by a simple application of strong alkaline solution on wax paper. Under optimal circumstances, the Cu@Cu 2 S@N–C electrochemical biosensor exhibits broad detection range of 2–7500 μM (R2 = 0.996) with low detection limit of 0.16 μM (S/N = 3) and high sensitivity of 1996 μA mM−1 cm−2. Additionally, the constructed μ-PAD also exhibited excellent selectivity, stability, and reproducibility. By rationally designing the double-shell hollow nanostructure and introducing Cu-encapsulated inner layer, the synthesized Cu@Cu 2 S@N–C hollow nanocubes show large specific surface area, short diffusion channels, and high stability. The proposed origami μ-PAD has been successfully applied to serum samples without any additional sample preparation steps for glucose determination, offering a new perspective for early nonenzymatic glucose diagnosis. [Display omitted] • Cu@Cu 2 S@N–C hollow nanocubes-based origami μ-PAD sensor was fabricated. • Cu@Cu 2 S@N–C hollow nanocubes possessed large specific surface area and high stability. • Cu@Cu 2 S@N–C hollow nanocubes exhibited excellent electrocatalytic performance for glucose. • The proposed sensor was applied for glucose detection without any additional sample preparation steps. [ABSTRACT FROM AUTHOR]

Fluid control on a paper channel is necessary for analysis with multiple reagents, such as enzyme-linked immunosorbent assay (ELISA) in microfluidic paper-based analytical devices (µPADs). In this study, a thermo-responsive valve was fabricated by polymerizing N-isopropylacrylamide on a PVDF porous membrane by plasma-induced graft polymerization. The polymerized membrane was observed by scanning electron microscopy (SEM), and it was confirmed that more pores were closed at temperatures below 32 °C and more pores were opened at temperatures above 32 °C. Valve permeability tests confirmed that the proposed polymerized membrane was impermeable to water and proteins at temperatures below 32 °C and permeable to water at temperatures above 32 °C. The valve could also be reversibly and repeatedly opened and closed by changing the temperature near 32 °C. These results suggest that plasma-induced graft polymerization may be used to produce thermo-responsive valves that can be opened and closed without subsequent loss of performance. These results indicate that the thermo-responsive valve fabricated by plasma-induced graft polymerization could potentially be applied to ELISA with µPADs. [ABSTRACT FROM AUTHOR]

Microfluidic paper-based analytical devices (µPADs) represent one of the promising green analytical strategies for low-cost and simple determination of various analytes. The actual task is the development of such devices for quantitation of antioxidants, e.g., flavonoids. In this paper, possibilities of a novel three-reagent µPAD including silver nitrate, 4-nitrophenyldiazonium tetrafluoroborate, and iron(III) chloride as reagents are assessed with respect to the determination of dihydroquercetin. It is shown that all the three reagents produce different colorimetric responses that can be detected by a mini-spectrophotometer–monitor calibrator or by a smartphone. The method is applicable to direct measuring high contents of dihydroquercetin (the linearity range is 0.026–1 mg mL−1, and the limit of detection is 7.7 µg mL−1), which is favorable for many dietary supplements. The analysis of a food supplement was possible with the relative standard deviations of 9–26%, which is satisfactory for quantitative and semiquantitative determinations. It was found that plotting a calibration graph in 3D space of the three reagents' responses allows us to distinguish dihydroquercetin from its close structural analogue, quercetin. [ABSTRACT FROM AUTHOR]

Microfluidic paper‐based analytical devices (μPADs) are commonly used in point‐of‐care testing (POCT) due to their advantages of low cost and easy operation. However, the fabrication and potential environmental pollution of μPADs are the universal and pendent concerns. In this respect, a simple and novel method is developed for preparing photo‐adjustable μPADs using the photocatalytic properties of TiO2. A superhydrophobic TiO2‐paper substrate with unique photoactivity is fabricated. The flow channels on the superhydrophobic TiO2‐paper substrate can be quickly generated and on‐demand edited by a simple UV irradiation process, to fabricate various 2D and 3D μPADs for specific applications. After usage, the residual organics and biological specimens on μPADs can be easily bleached via TiO2‐assited degradation under UV or sunlight, avoiding the risk of potential environment pollution and disease transmission. The successful application of such novel substrate in the fabrication of smart and safe μPADs for sweat analysis and motion monitoring is shown. [ABSTRACT FROM AUTHOR]

Abstract Microfluidic paper-based analytical devices (μPADs) for separating particles have been playing a key role for point-of-care diagnostics in the area of remote settings. While splendid separation methods using μPADs have been explosively developed, they still require external devices inducing external field. In this work, the spontaneous separation method in μPADs was suggested by leveraging convective flow (the imbibition of paper and nanoporous medium) and diffusiophoresis by ion exchange medium. Especially, the paper’s fast imbibition was utilized as driving particles at the first stage, which results in fast overall processing in contrast to the spontaneous separation method of microfluidic chip integrated with only ion exchange medium. Therefore, our novel spontaneous selective preconcentration method based on μPADs would have key potential to be used in portable point-of-care devices in remote settings.

Chlorpyrifos (CHL), profenofos (PRO) and cypermethrin (CYP) are widely used in combination to increase crop yields. However, these three pesticides can cause serious harm to human health and do not easily degrade. In this study, a novel visible paper sensor has been prepared successfully and different colorimetric reactions were utilized to detect the three pesticides simultaneously. The sensor was constructed by grafting a zwitterionic polymer onto a cellulose filter (CF) and placing it on a glass surface modified with PDMS. The branch shape was designed to form multiple detection areas, which were modified with specific pesticides and corresponding chromogenic reagents. The as-prepared colorimetric platform exhibited high sensitivity, a short detection time, a good linear response and a low detection limit (LOD) for the three pesticides (chlorpyrifos: y = 46.801 − 1.939x, R2 = 0.983, LOD = 0.235 mg/L; profenofos: y = 40.068 + 42.5x, R2 = 0.988, LOD = 4.891 mg/L; cypermethrin: y = 51.993 + 1.474x, R2 = 0.993, LOD = 4.053 mg/L). The comparison of the results obtained by the proposed paper sensor and those obtained by spectrophotometry further revealed the stability and reliability of the paper sensor. In particular, the color intensity of the interaction between the pesticides and coloring agents could be directly observed by the human eye. The consistency of the colorimetric/optical assay was proven in real target pesticide samples. Thus, this sensing strategy provides a portable, cost-effective, accurate and visualized paper platform, which could be suitable for application in the fruit and vegetable industry for monitoring CHL, PRO and CYP in parallel.

Herein, we describe the development of a paper-based device to detect nucleic acids of pathogens of interest in complex samples using loop-mediated isothermal amplification (LAMP) by producing a colorimetric response visible to the human eye. To demonstrate the utility of this device in emerging public health emergencies, we developed and optimized our device to detect SARS-CoV-2 in human saliva without preprocessing. The resulting device was capable of detecting the virus within 60 min and had an analytical sensitivity of 97% and a specificity of 100% with a limit of detection of 200 genomic copies/μL of patient saliva using image analysis. The device consists of a configurable number of reaction zones constructed of Grade 222 chromatography paper separated by 20 mil polystyrene spacers attached to a Melinex® backing via an ARclean® double-sided adhesive. The resulting device is easily configurable to detect multiple targets and has the potential to detect a variety of pathogens simply by changing the LAMP primer sets.

Paper-based microfluidic analytical devices (μPADs) have shown great potential in the field of analysis due to their advantages of rapid analysis, environmental friendliness and the ability to realize the flow of fluid without external power. Saliva is an emerging biofluid which is used in diseases diagnostic and screening for the easy collection and the reflection of the physiological state. This review focuses on the fabrication methods for two-dimensional (2D) and three-dimensional (3D) μPADs and their applications on the saliva analysis. In the first part, the flow mechanism in μPADs is discussed. The second part mainly introduces the fabrication methods for the μPADs and compares the different methods. The third part presents the application of μPADs in the detection of biomarkers such as nitrite, glucose, and thiocyanate in saliva. Finally, the research directions of saliva analysis are discussed in the conclusion. There have been a lot of researches on μPADs, but the fabrication methods and applications need to be further studied to meet the commercial needs. [ABSTRACT FROM AUTHOR]

Microfluidic paper-based analytical devices (µPADs) are leading the field of low-cost, quantitative in-situ assays. However, understanding the flow behavior in cellulose-based membranes to achieve an accurate and rapid response has remained a challenge. Previous studies focused on commercial filter papers, and one of their problems was the time required to perform the test. This work studies the effect of different cellulose substrates on diffusion-based sensor performance. A diffusion-based sensor was laser cut on different cellulose fibers (Whatman and lab-made Sisal papers) with different structure characteristics, such as basis weight, density, pore size, fiber diameter, and length. Better sensitivity and faster response are found in papers with bigger pore sizes and lower basis weights. The designed sensor has been successfully used to quantify the ionic concentration of commercial wines with a 13.6 mM limit of detection in 30 s. The developed µPAD can be used in quantitative assays for agri-food applications without the need for any external equipment or trained personnel.

A user-friendly, low-cost detector able to read the RGB indexes of microfluidic paper-based analytical devices (µPADs) was developed. The RGB-detector was built with 3D printing using PLA+ and reused Li-ion batteries. It is Arduino-based, which provides an easy interface between the sensor TCS3200, which reads the quadratic wave of the times corresponding to the RGB numbers, the Arduino itself, whose software translates the times into RGB values, and the touchscreen display, NX3224T028, which shows the results. This detector permits multi-sample analysis since it has a sample holder that can keep up to six µPADs simultaneously and rotate after the display’s request. This work shows how the readings of the RGB indexes by the proposed RGB-detector implement the measurements’ reproducibility. As a proof-of-concept, the RGB-detector application to a green array of µPADs for pH measurement coupled with chemometric analysis allowed us to achieve good results in terms of precision and agreement with the pH values measured by a classical pH-meter.

We developed a portable device for blood detection as well as blood typing assay, using a microfluidic paper-based analytical devices (µPADs) which proves to be sensitive, cost-effective, user-friendly and field-appropriate. A different patterned µPAD was fabricated using wax and a laser printer for detection of blood and ABO and Rh blood typing. Five different blood detection tests and two blood typing systems, i.e. ABO and Rh, were performed within minutes without following any complex protocol and were simple to infer. The performance of this hand-held μPAD proves it to be a reliable alternative to lab-based tests, at a fraction of the cost and with simplicity. The developed μPAD provides on-site blood detection as well as blood typing, which proves to be a reasonable and very useful add-on to conventional forensic and clinical investigations. [ABSTRACT FROM AUTHOR]

Microfluidic analytical devices manufactured on paper and similar inexpensive substrates (µ-PADs) have shown considerable promise for disease diagnostics in resource-limited regions. However, current commercialization approaches can be improved substantially by addressing existing technical challenges associated with µ-PADs. Among these, off-device plasma separation from whole blood is a critical challenge in µ-PAD technology that limits commercialization. Existing µ-PADs made by combining multiple components require extra fabrication steps and manufacturing material. Our approach utilizes a two-step plasma process to fabricate single-layer semi-enclosed µ-PADs directly on a commercially available blood plasma separation membrane to incorporate blood plasma separation functionality into the device. The semi-enclosed µ-PADs are bonded with low-cost adhesive plastic tape to provide mechanical support to the device and make it more mechanically robust for field applications. Detection zones of the µ-PADs have also been modified with a cellulose nanocrystal (CNC) to increase colorimetric signal homogeneity, thus enhancing signal quality. The CNC-modified µ-PADs have been used for colorimetric detection of two model analytes (glucose and albumin) in whole blood. Colorimetric signals for both glucose and albumin from whole blood samples were consistent with the calibration curves generated using stock solutions. [ABSTRACT FROM AUTHOR]

[Display omitted] • An enrichment method of glycoproteins using microfluidic paper-based analytical devices (μPADs) was developed. • The μPADs consist of two separate parts: the enrichment part (Chip I) and the detection part (Chip II). • The 3-aminophenylboronic acid immobilized on Chip I can capture glycoproteins at pH 8.0 and release them at pH 3.0. • The method can effectively enrich an ovalbumin solution with concentrations as low as 1.0 × 10−11 mol/L. • Glycoproteins was enriched from 1 × 106-fold diluted real egg white samples. We introduce a facile method for glycoprotein enrichment, separation, and detection using microfluidic paper-based analytical devices (μPADs). The μPADs comprise two distinct components: the enrichment region (Chip I) and the detection section (Chip II). The reusable Chip I is fabricated by immobilizing 3-aminophenylboronic acid (APBA) onto a circular paper substrate, demonstrating exceptional glycoprotein affinity. In disposable Chip II, glycoproteins react with periodic acid, generating carbonyl compounds that produce a purple-colored product upon reacting with the Schiff reagent. Alterations in color intensity are captured using an ordinary cell phone and are subsequently analyzed using Photoshop software. Observations note a linear increase in average color intensities from 1.0 × 10−3 to 5.0 × 10−3 mol/L, with an R2 value of 0.9927. The relative standard deviations (RSDs) of inter-day (n = 5) and intra-day (n = 5) assessments stand at 1.5 % and 1.4 %, respectively. Our proposed approach can effectively enrich an ovalbumin (OVA) solution with concentrations as low as 1.0 × 10−11 mol/L, resulting in enrichment rates of 107 to 108 times. The interference deviations of five substances relative to OVA lie within the range of − 2.0 % to + 2.0 %. The methodology successfully enriches glycoproteins from egg white samples diluted by a factor of 1 × 106, demonstrating that the proposed method is remarkably suitable for enriching glycoproteins from authentic samples. [ABSTRACT FROM AUTHOR]

[Display omitted] • Orientationally immobilized enzyme microreactor (OIMER) was developed. • OIMER embedded microfluidic paper-based analytical devices was prepared. • The method is simple, accurate, low cost, rapid and effective in pesticide detection. • The method avoids the need for sophisticated instruments. • The system was applied to detect pesticide residues in vegetable samples. Orientationally immobilized enzyme microreactors (OIMERs), embedded in microfluidic paper-based analytical devices (μPADs) were developed for improved detection of pesticide residues in food. Acetylcholinesterase (AChE) was orientationally immobilized on the reusable Part I of the μPADs, using the specific affinity binding of concanavalin A (Con A) to a glycosyl group on AChE. Using the disposable Part II, facile colorimetric quantification was performed with a smartphone and software, or qualitative detection by a naked-eye visual test. The AChE immobilized in OIMERs not only had improved activity and stability, but also high sensitivity, with a limit of detection as low as (0.007 ± 0.003) μg/mL. The method was used to detect pesticides residues in real vegetable samples; the recovery (88.6–102.7%) showed high reliability for pesticide residues detection in foods. A molecular docking study and an enzyme kinetic analysis were conducted to characterize the mechanism of action of the OIMERs. [ABSTRACT FROM AUTHOR]

Fluid control on a paper channel is necessary for analysis with multiple reagents, such as enzyme-linked immunosorbent assay (ELISA) in microfluidic paper-based analytical devices (µPADs). In this study, a thermo-responsive valve was fabricated by polymerizing N-isopropylacrylamide on a PVDF porous membrane by plasma-induced graft polymerization. The polymerized membrane was observed by scanning electron microscopy (SEM), and it was confirmed that more pores were closed at temperatures below 32 °C and more pores were opened at temperatures above 32 °C. Valve permeability tests confirmed that the proposed polymerized membrane was impermeable to water and proteins at temperatures below 32 °C and permeable to water at temperatures above 32 °C. The valve could also be reversibly and repeatedly opened and closed by changing the temperature near 32 °C. These results suggest that plasma-induced graft polymerization may be used to produce thermo-responsive valves that can be opened and closed without subsequent loss of performance. These results indicate that the thermo-responsive valve fabricated by plasma-induced graft polymerization could potentially be applied to ELISA with µPADs.

Microfluidic paper-based analytical devices (µPADs) represent one of the promising green analytical strategies for low-cost and simple determination of various analytes. The actual task is the development of such devices for quantitation of antioxidants, e.g., flavonoids. In this paper, possibilities of a novel three-reagent µPAD including silver nitrate, 4-nitrophenyldiazonium tetrafluoroborate, and iron(III) chloride as reagents are assessed with respect to the determination of dihydroquercetin. It is shown that all the three reagents produce different colorimetric responses that can be detected by a mini-spectrophotometer–monitor calibrator or by a smartphone. The method is applicable to direct measuring high contents of dihydroquercetin (the linearity range is 0.026–1 mg mL−1, and the limit of detection is 7.7 µg mL−1), which is favorable for many dietary supplements. The analysis of a food supplement was possible with the relative standard deviations of 9–26%, which is satisfactory for quantitative and semiquantitative determinations. It was found that plotting a calibration graph in 3D space of the three reagents’ responses allows us to distinguish dihydroquercetin from its close structural analogue, quercetin.

A method is described for cysteine (Cys) determination on paper-based analytical devices using aspartic acid modified gold nanoparticles (Asp-AuNPs). The Asp-AuNPs were characterized by their size, zeta potential, and UV-visible absorption spectrum. After the addition of Cys, it will interact with Asp-AuNPs selectively and leads to the aggregation of Asp-AuNPs. A color change from red to blue can be observed on the paper-based analytical devices. The results were recorded using a cell phone and subsequently analyzed using the Photoshop software. The ratiometric color intensity at red channel and blue channel (Red/Blue) increased linearly in the range 99.9–998.7 μM for Cys (R = 0.9984), and the limit of detection was 1.0 μM. The effects of assay conditions have been investigated and are discussed. The Cys concentration was determined as (0.27 ± 0.02 mM) in human plasma, and the recovery was from 99.2 to 101.1%. [ABSTRACT FROM AUTHOR]

The diagnosis of dengue infection is still a critical factor determining success in the clinical management and treatment of patients. Here, the development of microfluidic paper-based analytical devices (μPADs) utilizing a sandwich immunoassay on wax patterned paper functionalized with anti-dengue NS1 monoclonal antibodies for point-of-care detection of dengue NS1 (DEN-NS1-PAD) is reported. Various assay conditions, including the length of the channel and diluent, were optimized, and the response detected by the naked eye and digitized images within 20–30 min. The DEN-NS1-PAD was successfully tested in the field for detecting dengue NS1 in buffer, cell culture media, and human serum. The limit of detection (LoD) of the DEN-NS1-PAD obtained with the naked eye, scanner, and a smartphone camera was 200, 46.7, and 74.8 ng mL−1, respectively. The repeatability, reproducibility, and stability of the DEN-NS1-PAD were also evaluated. High true specificity and sensitivity in the serum of pediatric patients were observed. These evaluation results confirm that the DEN-NS1-PAD can potentially be used in point-of-care dengue diagnostics, which can significantly impact on the spreading of mosquito-borne diseases, which are likely to become more prevalent with the effects of global warming. [ABSTRACT FROM AUTHOR]

Microfluidic paper-based analytical devices (μPADs) for separating particles have been playing a key role for point-of-care diagnostics in the area of remote settings. While splendid separation methods using μPADs have been explosively developed, they still require external devices inducing external field. In this work, the spontaneous separation method in μPADs was suggested by leveraging convective flow (the imbibition of paper and nanoporous medium) and diffusiophoresis by ion exchange medium. Especially, the paper's fast imbibition was utilized as driving particles at the first stage, which results in fast overall processing in contrast to the spontaneous separation method of microfluidic chip integrated with only ion exchange medium. Therefore, our novel spontaneous selective preconcentration method based on μPADs would have key potential to be used in portable point-of-care devices in remote settings. [ABSTRACT FROM AUTHOR]

The incompatibility of most organic solvents with acetylcholinesterase (AChE) inhibition assay normally limits pesticide extraction efficiency in sample pretreatment, which might cause false negatives in real world sample assessment. Herein, a novel method has been developed for an improved AChE inhibition assay via organic solvent extraction combined spontaneous in situ solvent evaporation on microfluidic paper-based analytical devices. Enzyme pre-immobilization procedure was spared and AChE was added to the system after sampling step until a complete in-situ solvent evaporation process was performed on chip. IC 50 levels of the six investigated organophosphate and carbamate pesticides indicated a completely eliminated influence of solvents on AChE behavior with the new method. Most importantly, analytical performances were significantly improved in food sample measurements. Reduction in matrix effect was observed when acetonitrile was adopted for lettuce sample pretreatment instead of water. Studies on different pesticides suggested a remarkably decreased discrimination effect on recoveries from sample pretreatment with the new developed method. The recovery level for phoxim spiked head lettuce samples reached (107.5 ± 14.2) %, in comparison with that of (18.6 ± 1.4) % from water-based extraction. Spiked water and apple juice samples with carbaryl concentration of as low as 0.02 mg L−1 were also successfully recognized with the present method by visual detection. This is the first report on direct sampling of organic extracts for AChE inhibition assay on-chip and it might provide a new perspective for real world sample assessments involving bio-reagents. Image 1 • An AChE inhibition assay utilizing organic solvents for pesticide extraction in food analysis. • Online solvent elimination via spontaneous evaporation before contacting with enzyme. • Significantly improved recoveries for different pesticides in food samples. • Rapid and direct pesticide pre-enrichment ability during sample extraction. [ABSTRACT FROM AUTHOR]

A major goal of green chemistry is to avoid the use of toxic organic solvents. In particular, there are few green methods allowing extraction and quantification of chromium species in environmental samples. Here, we developed home-made, dual-gel electromembrane extraction combined with a microfluidic paper-based analytical device for the simultaneous determination of Cr(III) and Cr(VI) in water, without using any organic solvent. The positively charged Cr(III) and negatively charged Cr(VI) migrated selectively into the cathodic (pH 2.0) and anodic (pH 3.0) aqueous acceptor phases, respectively. After extraction, the anodic acceptor phase containing Cr(VI) was analyzed directly by a microfluidic paper-based analytical device, after the addition of the diphenylcarbazide colorimetric reagent. The cathodic acceptor phase containing Cr(III) was mixed with Ce(IV) to oxidize Cr(III) to Cr(VI), and then, Cr(VI) ions were detected on the microfluidic paper-based analytical device after adding diphenylcarbazide. Under the optimized conditions, limits of detection of 2.0 and 3.0 ng/mL and extraction recoveries of 58.8% and 83.3% were achieved for Cr(VI) and Cr(III), respectively. [ABSTRACT FROM AUTHOR]

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.

Microfluidic paper-based analytical devices (µPADs) are a promising technology to enable accurate and quantitative in situ assays. Paper’s inherent hydrophilicity drives the fluids without the need for external pressure sources. However, controlling the flow in the porous medium has remained a challenge. This study addresses this problem from the nature of the paper substrate and its design. A computational fluid dynamic model has been developed, which couples the characteristics of the porous media (fiber length, fiber diameter and porosity) to the fluidic performance of the diffusion-based µPAD sensor. The numerical results showed that for a given porous membrane, the diffusion, and therefore the sensor performance is affected not only by the substrate nature but also by the inlets’ orientation. Given a porous substrate, the optimum performance is achieved by the lowest inlets’ angle. A diffusion-based self-referencing colorimetric sensor was built and validated according to the design. The device is able to quantify the hydronium concentration in wines by comparison to 0.1–1.0 M tartaric acid solutions with a 41.3 mM limit of detection. This research showed that by proper adjustments even the simplest µPADs can be used in quantitative assays for agri-food applications.

In this study, we developed and compared three different methods for chromium speciation in water samples using microfluidic paper-based analytical devices (μPADs). In all methods, detection was based on the complexation reaction of Cr(VI) with diphenylcarbazide on the μPADs. Cr(III) ions were oxidized to Cr(VI) by Ce(IV) prior to colorimetric detection on the μPADs. In the first method, oxidization of Cr(III) to Cr(VI) in the solution containing both trivalent and hexavalent chromium was performed using a batch procedure to obtain total chromium. A dual electromembrane extraction (DEME) technique for simultaneous preconcentration and extraction of chromium species and a single electromembrane extraction (SEME) for preconcentration and extraction of Cr(VI)/total chromium [quantified as Cr(VI) content after oxidation of Cr(III) ions to Cr(VI)] were used in the second and third methods, respectively. The electromembrane extraction was based on the electrokinetic migration of cationic Cr(III) and anionic Cr(VI) toward the cathode and anode, respectively, into the two different hollow fibres. Octanol-1 and bis(2-ethylhexyl) phosphate (DEHP) in octanol-1 (0.7% v/v) were the most suitable supported liquid membranes for extraction of Cr(VI) and Cr(III), respectively. Among these methods, SEME showed the lowest limits of detection for both analytes. Under optimized conditions, linear calibrations were obtained for Cr(III) from 3 to 30 μg L−1 and for Cr(VI) from 3 to 70 μg L−1. The detection limits were 1.0 μg L−1 and 0.7 μg L−1 for Cr(III) and Cr(VI), respectively. Our developed method was applied to analyse water samples spiked with different concentrations of Cr(III) and Cr(VI) at the parts-per-billion (ppb) level. The statistical evaluation showed that the proposed method agreed well with the validation method, i.e., inductively coupled plasma atomic emission spectroscopy (ICP-AES). Image 1 • Electromembrane was used for sensitivity enhancement of μPAD's limit of detection. • 1,5-diphenylcarbazide (DPC) was used as selective colorimetric reagent for detection of Cr (VI) on paper-based device. • Cr(III) was oxidized to Cr(VI) by Ce(IV) in acidic medium. • The limit of detection of 1.0 μg L−1 and 0.7 μg L−1 for Cr(III) and Cr(VI), respectively, were obtained [ABSTRACT FROM AUTHOR]

Microfluidic paper-based analytical devices (μPADs) for detection of hydrogen peroxide and glucose have been developed. The analytical performance of colorimetric detection using the conventional starch-iodine color reaction has been significantly improved by using gelatin as the surface modifier which retains the enzyme activity in the dry filter paper strip, improves antioxidability, as well as decreases the strong background signal. Under optimal conditions, the color intensities show a good linear relationship with glucose concentration ranging from 0.5 to 5 mM and hydrogen peroxide concentration from 0.5 to 6 mM, with the detection limit of 0.05 mM and 0.1 mM, respectively. In addition, the accuracy of colorimetric sensor has been successfully assessed in detecting glucose from real human serum samples and recovery value ranges from 95.7% to 97%, which are approaching to the glucose oxidase endpoint. The new colorimetric assay exhibits high sensitivity, good selectivity, acceptable stability and reproducibility. The present approach is promising for monitoring glucose for point of care diagnostic applications, especially in regions with resource-limited settings. Image 1 • A facile and sensitive colorimetric assay was proposed based on μPADs coupled with starch-iodide-gelatin system. • The gelatin can provide biocompatible microenvironment and improve the analytical performance of colorimetric methods. • The accuracy of colorimetric sensor was successfully assessed to detect the glucose in the real serum samples. • This colorimetric assay is used to monitor glucose for point of care diagnostic applications. [ABSTRACT FROM AUTHOR]

Microfluidic paper-based devices (μPADs) and wearable devices have been highly studied to be used as diagnostic tools due to their advantages such as simplicity and ability to provide instrument-free fast results. Diseases such as periodontitis and diabetes mellitus can potentially be detected through these devices by the detection of important biomarkers. This study describes the development of μPADs through craft cutter printing for glucose and nitrite salivary diagnostics. In addition, the use of μPADs integrated into a mouthguard as a wearable sensor for glucose monitoring is also presented. μPADs were designed to contain two detection zones for glucose and nitrite assays and a sampling zone interconnected by microfluidic channels. Initially, the analytical performance of the proposed μPADs was investigated and it provided linear behavior (r2 ≥ 0.994) in the concentration ranges between 0 to 2.0 mmol L−1 and 0 to 400 μmol L−1 for glucose and nitrite, respectively. Under the optimized conditions, the limits of detection achieved for glucose and nitrite were 27 μmol L−1 and 7 μmol L−1, respectively. Human saliva samples were collected from healthy individuals and patients previously diagnosed with periodontitis or diabetes and then analyzed on the proposed μPADs. The results found using μPADs revealed higher glucose concentration values in saliva collected from patients diagnosed with diabetes mellitus and greater nitrite concentrations in saliva collected from patients diagnosed with periodontitis, as expected. The results obtained on μPADs did not differ statistically from those measured by spectrophotometry. With the aim of developing paper-based wearable sensors, μPADs were integrated, for the first time, into a silicone mouthguard using a 3D-printed holder. The proof of concept was successfully demonstrated through the monitoring of the glucose concentration in saliva after the ingestion of chocolate. According to the results reported herein, paper-based microfluidic devices offer great potential for salivary diagnostics, making their integration into a silicone mouthguard possible, generating simple, low-cost, instrument-free, and powerful wearable sensors. [ABSTRACT FROM AUTHOR]

Highlights • A sample-in-answer-out assay is developed for detection of sub-nanomolar Ag+. • It only requires a cheap timer to realize instrument-free quantitative measurement. • A target-triggered autocatalysis strategy is proposed for signal amplification. • Nanopore membrane-integrated paper devices serve as portable testing platforms. Abstract This work initially describes a sample-in-answer-out paper-based assay for simple, rapid, sensitive, specific, and low-cost detection of silver (I) (Ag+) ions. It combines Ag+-triggered in situ autocatalysis for signal amplification with an instrument-free, timing readout in 3D origami paper-based analytical devices integrated with nanopore membranes (NMs). The autocatalytic amplification is based on the oxidability of Ag+ towards 3,3',5,5'-tetramethylbenzidine (TMB) loaded in the NMs. The Ag+ ions are rapidly reduced by the TMB to form silver nanoparticles that further act as nanocatalysts to catalyse the oxidation of more TMB molecules to their derivatives by Ag+ ions for producing bigger (and more) metal particles. The NM's pores are thus partially blocked by these metal particles to adjust (prolong) the flowing time of a coloured reagent in the paper device. Under optimized conditions, this developed point-of-need method could quantitatively detect the Ag+ ions in a linear concentration range of 1.9–62.5 nM within 4 min, by simply using a ubiquitous phone (or a timer). Its detection limit was estimated to be ˜0.91 nM according to the 3σ rule. The recoveries of analyzing real water examples (i.e., pond water, tap water, drinking water, and soil water) were from ˜95.2 to ˜103.9%, and the corresponding relative standard deviations ranged from 3.3 to 8.1% (n = 6). [ABSTRACT FROM AUTHOR]

Abstract We report the colorimetric detection of dopamine (DA) on microfluidic paper-based analytical devices (μPADs) using an oxidation-reduction method. Here, dopamine reacts with ferric chloride forming reduced Fe2+ that subsequently reacts with phenanthroline to form the red tris(1,10-phenanthroline)iron(II) complex. The devices were fabricated by wax printing and changes in color intensity were recorded using a common cell phone. Subsequent analysis using Photoshop software, yielded a limit of detection (LOD) for DA of 0.37 μmol/L with a linear range of 0.527–4.75 μmol/L and relative standard deviation of 0.11% (inter-day) and 0.15% (intra-day) for n = 15 paper chips. The effects of detection conditions have been investigated and are discussed. Cow serum samples and human blood serum and plasma samples were detected. The work, herein, demonstrates the potential of this method as a low cost and rapid colorimetric technique to detect DA in real samples. Graphical abstract fx1 Highlights • A colorimetric detection of dopamine (DA) on microfluidic paper-based analytical devices (μPADs) was developed. • The changes in color intensity were recorded using a common cell phone and analysis using Photoshop software. • The method provide an inexpensive and selective determination of DA with low LOD. It does not need other assistances. • The relative standard deviation (RSD) of inter-day (n =15) and intra-day (n =15) is no more than 0.2%. • Cow serum samples and human serum and plasma samples were detected. [ABSTRACT FROM AUTHOR]

Abstract A simple and highly sensitive procedure based on the combination of hollow fiber membrane liquid phase microextraction and a microfluidic paper-based analytical device (µPAD) was developed for pre-concentration and determination of hexavalent chromium in water samples. The hexavalent chromium was pre-concentrated using the HF-LPME technique via ion exchange or a coupled transport process through a supported ionic liquid (Aliquat 336) prior to colorimetric detection with diphenylcarbazide on the µPAD. The violet colour could be seen by the naked eye. Images from the µPADs were scanned using a commercial desktop scanner at 600 dpi resolution. ImageJ software was used for quantitative analysis by measuring the intensity values at green colour channel since it gave the best sensitivity among the RGB colour. Under optimal conditions, the calibration curve was linear in the range 10–90 µg L-1, with a limit of detection of 3 µg L-1. The developed method was successfully applied to determine the level of hexavalent chromium spiked into natural water samples at the parts- per -billion (ppb) level, and the results were in good agreement with those obtained using inductively coupled plasma atomic emission spectroscopy (ICP-AES). The developed method was able to improve the detection limit of the conventional µPAD, and was expected to be used for the effective analysis of hexavalent chromium in natural water. Graphical abstract fx1 Highlights • HF-LPME was used for sensitivity enhancement of paper-based analytical device for detection of Cr (VI) in water samples. • 1,5-diphenylcarbazide (DPC) was used as colorimetric reagent for detection of Cr (VI) on paper-based device. • The methodology showed good reproducibility with low operational cost. • The limit of detection of 3 µg L-1 was obtained. [ABSTRACT FROM AUTHOR]

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.

A fully integrated device for salivary detection with a sample-in-answer-out fashion is critical for noninvasive point-of-care testing (POCT), especially for the screening of contagious disease infection. Microfluidic paper-based analytical devices (μPADs) have demonstrated their huge potential in POCT due to their low cost and easy adaptation with other components. This study developed a generic POCT platform by integrating a centrifugal microfluidic disc with μPADs to realize sample-to-answer salivary diagnostics. Specifically, a custom centrifugal microfluidic disc integrated with μPADs is fabricated, which demonstrated a high efficiency in saliva treatment. To demonstrate the capability of the integrated device for salivary analysis, the SARS-CoV-2 Nucleocapsid (N) protein, a reliable biomarker for SARS-CoV-2 acute infection, is used as the model analyte. By the chemical treatment of the μPAD surface, and by optimizing the protein immobilization conditions, the on-disc μPADs were able to detect the SARS-CoV-2 N protein down to 10 pg mL -1 with a dynamic range of 10-1000 pg mL -1 and an assay time of 8 min. The integrated device was successfully used for the quantification of the N protein of pseudovirus in saliva with high specificity and demonstrated a comparable performance to the commercial paper lateral flow assay test strips.

Abstract This review reports on recent advances in state-of-the-art detection methods for microfluidic paper-based analytical devices (μPADs). The review commences by describing the materials, fabrication methods and driving mechanisms employed in μPADs. The review then explores the main detection methods and applications of μPADs proposed over the past five years. The discussions focus on seven detection technologies, namely (1) colorimetric detection, (2) fluorescent detection, (3) electrochemical detection and photoelectrochemical detection, (4) chemiluminescent detection, (5) electrochemiluminescent detection, (6) nanoparticle-based detection, and (7) spectrometry detection. Overall, the review provides a useful insight into recent advances in the μPAD detection field and serves as a useful source of reference for further research and innovation in the field. Highlights • Detection methods of microfluidic paper-based analytical devices (μPADs) have been discussed. • The review focuses on proposals for main detection methods and applications of μPADs. • A comprehensive review of the main applications of μPADs over the past five years. [ABSTRACT FROM AUTHOR]

Abstract: Isoelectric focusing plays a critical role in the analysis of complex protein samples. Conventionally, isoelectric focusing is implemented with carrier ampholytes in capillary or immobilized pH gradient gel. In this study, we successfully exhibited a carrier ampholyte‐free isoelectric focusing on paper‐based analytical device. Proof of the concept was visually demonstrated with color model proteins. Experimental results showed that not only a pH gradient was well established along the open paper fluidic channel as confirmed by pH indicator strip, the pH gradient range could also be tuned by the catholyte or anolyte. Furthermore, the isoelectric focusing fractions from the paper channel can be directly cut and recovered into solutions for post analysis with sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and matrix‐assisted laser desorption/ionization‐time‐of‐flight mass spectrometry. This paper‐based isoelectric focusing method is fast, cheap, simple and easy to operate, and could potentially be used as a cost‐effective protein sample clean‐up method for target protein analysis with mass spectrometry. [ABSTRACT FROM AUTHOR]

A visual colorimetric microfluidic paper-based analytical device (μPAD) was constructed following the direct synthesis of enzyme-inorganic hybrid nanomaterials on the paper matrix. An inorganic solution of MnSO 4 and KH 2 PO 4 containing a diluted enzyme (glucose oxidase, GOx) was subsequently pipetted onto cellulose paper for the in situ growth of GOx@Mn 3 (PO 4 ) 2 hybrid functional materials. The characterization of the morphology and chemical composition validated the presence of hybrid materials roots in the paper fiber, while the Mn 3 (PO 4 ) 2 of the hybrid provided both a surface for enzyme anchoring and a higher peroxidase-like catalytic activity as compared to the Mn 3 (PO 4 ) 2 crystal that was synthesized without enzyme modulation. This new approach for the in situ growth of an enzyme-inorganic hybrid on a paper matrix eliminates centrifugation and the dry process by casting the solution on paper. The sensing material loading was highly reproducible because of the accuracy and stability of pipetting, which eventually contributed to the reliability of the μPAD. The self-assembled natural and artificial enzyme hybrid on the μPADs specifically detected glucose from a group of interferences, which shows great specificity using this method. Moreover, the colorimetric signal exhibited detection limitation for glucose is 0.01 mM, which lies in the physiological range of glucose in biological samples. [ABSTRACT FROM AUTHOR]