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Heavy metal contamination is an increasing global threat to human and environmental health, particularly in resource-limited areas. Traditional platforms for heavy metal detection are labor intensive and expensive and require lab facilities. While paper-based colorimetric sensors offer a simpler approach, their sensitivity limitations prevent them from meeting legislative requirements for many metals. Existing preconcentration systems, on the other hand, can achieve lower detection limits but typically focus on analyzing only one metal, making comprehensive monitoring difficult. We address these limitations by introducing a low-cost preconcentration system coupled with colorimetric analysis for the simultaneous detection of seven metal ions at low ppb levels without the need for external equipment outside a smartphone. The system achieved detection limits of 15 ppb (Ni(II)), 7 ppb (Cu(II)), 2 ppb (Fe(III)), 20 ppb (Cr(VI)), 13 ppb (Pb(II)), 26 ppb (Hg(II)), and 15 ppb (Mn(II)) with six out of seven limits of detection values falling well below EPA regulatory guidelines for drinking water. The user-friendly Fill, Fold, Photo approach eliminates complex pretreatment steps. Smartphone-based detection offers portable quantification within seconds. Employing masking strategies ensured higher selectivity for each assay on the card, while our packaging protocols enable system stability for over 4 weeks of study, facilitating mass production and deployment within a realistic time frame. To validate the sensor's performance in real-world scenarios, the sensor was tested with environmental water samples. The sensor demonstrated good recovery, ranging from 77% to 94% compared to the standard ICP-MS method. Furthermore, spike recovery analysis confirmed the sensor's accuracy, with a relative standard deviation (RSD) of less than 15%. This technology holds significant promise for future development as a convenient, portable solution for field-based monitoring of a broad spectrum of water contaminants, including pesticides, PFAS, fertilizers, and beyond.

Rapid and inexpensive serological tests for SARS-CoV-2 antibodies are needed to conduct population-level seroprevalence surveillance studies and can improve diagnostic reliability when used in combination with viral tests. Here, we report a novel low-cost electrochemical capillary-flow device to quantify IgG antibodies targeting SARS-CoV-2 nucleocapsid proteins (anti-N antibody) down to 5 ng/mL in low-volume (10 μL) human whole blood samples in under 20 min. No sample preparation is needed as the device integrates a blood-filtration membrane for on-board plasma extraction. The device is made of stacked layers of a hydrophilic polyester and double-sided adhesive films, which create a passive microfluidic circuit that automates the steps of an enzyme-linked immunosorbent assay (ELISA). The sample and reagents are sequentially delivered to a nitrocellulose membrane that is modified with a recombinant SARS-CoV-2 nucleocapsid protein. When present in the sample, anti-N antibodies are captured on the nitrocellulose membrane and detected via chronoamperometry performed on a screen-printed carbon electrode. As a result of this quantitative electrochemical readout, no result interpretation is required, making the device ideal for point-of-care (POC) use by non-trained users. Moreover, we show that the device can be coupled to a near-field communication potentiostat operated from a smartphone, confirming its true POC potential. The novelty of this work resides in the integration of sensitive electrochemical detection with capillary-flow immunoassay, providing accuracy at the point of care. This novel electrochemical capillary-flow device has the potential to aid the diagnosis of infectious diseases at the point of care.

α 1 -Acid glycoprotein (AGP) is a glycoprotein present in serum, which is associated with the modulation of the immune system in response to stress or injuries, and a biomarker for inflammatory diseases and cancers. Here, we propose a pump-free microfluidic device for the electrochemical determination of AGP. The microfluidic device utilizes capillary-driven flow and a passive mixing system to label the AGP with the Os (VI) complex (an electrochemical tag) inside the main channel, before delivering the products to the electrode surface. Furthermore, thanks to the resulting geometry, all the analytical steps can be carried out inside the device: labeling, washing, and detection by adsorptive transfer stripping square wave voltammetry. The microfluidic device exhibited a linear range from 500 to 2000 mg L -1 ( R 2 = 0.990) and adequate limit of detection (LOD = 231 mg L -1 ). Commercial serum samples were analyzed to demonstrate the success of the method, yielding recoveries around 83%. Due to its simplicity, low sample consumption, low cost, short analysis time, disposability, and portability, the proposed method can serve as a point-of-care/need testing device for AGP.

The albumin-to-alkaline phosphatase ratio (AAPR) has been a cancer prognostic indicator. This paper presents the concept of a dual-color change distance-based paper device (dPAD) for albumin (Alb) and alkaline phosphatase (ALP) detection to evaluate this cancer prognostic index. Whereas Alb interacts with the bromocresol green (BCG) indicator to form a bluish-green complex, ALP hydrolyzes l-ascorbic acid-2-phosphate (AAP) to produce ascorbic acid (AA), which reacts with KIO 3 to generate I 2 and I - . I 2 /I - reacts with silver hexagonal nanoprisms (purple color) in the presence of Cu 2+ , resulting in a color change from purple to colorless. The distance of the color change from yellow to the bluish-green and purple to colorless correlates to Alb and ALP concentration, respectively. The angle index for the AAPR is then defined by drawing a straight line that connects the tops of the two changed band lengths in the detection area. The highest bluish-green color band length on the Alb region is the midpoint, which is the position set of the protractor at 0°, and the angle is measured using a simple protractor. The results indicate that an AAPR below 0.57 will have an angle greater than 40° and correlates with a risk factor for lung cancer. The naked-eye detection limits for Alb and ALP were found to be 0.8 g/L and 5 U/L ( n = 10), respectively. The practical application of the developed dPAD was successfully demonstrated by Alb and ALP analysis in human serum and validated against standard methods. The proposed method does not require incubation conditions for the ALP assay, which strongly reduces the overall analysis steps and time. Moreover, our device provides a low-cost, simple, sensitive, selective, accurate, and precise determination of the AAPR.

A paper-based method for heating preconcentration (PAD-HP) has been developed for the determination of Pb 2+ , Cd 2+ , Fe 3+ , and Ni 2+ . The design of our heating system was evaluated for dual quantification of ions using electrochemical and colorimetric methods simultaneously. The PAD-HP was used to detect Pb 2+ and Cd 2+ by anodic stripping voltammetry and to detect Fe 3+ and Ni 2+ by colorimetric reactions. Assay conditions were optimized by evaluating performance when changing the concentration of the colorimetric reagent, eluent volume, electrolyte concentration, and electrochemical parameters. Limits of detection (LOD) were determined to be 0.97 and 2.33 μg L -1 for Pb 2+ and Cd 2+ (via voltammetry) and 0.03 and 0.04 mg L -1 for Fe 3+ and Ni 2+ (via colorimetric assay), respectively. The relative standard deviations for assays were in the range of 5.76 to 10.12%. We observed that the PAD-HP method significantly enhanced the signal of all metals ions (14-100-fold, depending on the metal) in comparison to paper-based devices that did not use a heating preconcentration system. This PAD-HP method was successfully applied to the determination of metals ions in samples of drinking water, tap water, pond water, and wastewater. These results suggest that our approach can provide a convenient strategy to monitor aqueous samples for heavy metals with high sensitivity and selectivity.

Capillary forces are commonly employed to transport fluids in pump-free microfluidic platforms such as paper-based microfluidics. However, since paper is a porous material consisting of nonuniform cellulose fibers, it has some limitations in performing stable flow functions like mixing. Here, we developed a pump-free microfluidic device that enables rapid mixing by combining paper and plastic. The device was fabricated by laminating transparency film and double-sided adhesive and is composed of an overlapping inlet ending in a paper-based reaction area. The mixing performance of the developed device was confirmed experimentally using aqueous dyes and pH indicators. In addition, the absolute mixing index was evaluated by numerically calculating the concentration field across the microfluidic channels. To demonstrate the utility of the new approach, the detection of an organophosphate pesticide was carried out using a colorimetric enzymatic inhibition assay. The developed device and a smartphone application were used to detect organophosphate pesticide on food samples, demonstrating the potential for onsite analysis.

A simple and low-cost continuous-flow (CF) electrochemical paper-based analytical device (ePAD) coupled with thermoplastic electrodes (TPEs) was developed. The fast, continuous flow combined with flow injection analysis was made possible by adding two inlet reservoirs to the same paper-based hollow channel flowing over detection electrodes, terminating in a fan-shaped pumping reservoir. The upstream inlet reservoir was filled with buffer and provided constant flow through the device. Sample injections were performed by adding 2 μL of the sample to the downstream sample inlet. Differences in flow resistance resulted in sample plugs displacing buffer as the solution flowed over the working electrodes. The electrodes were fabricated by mixing carbon black and polycaprolactone (50% w/w). CF-TPE-ePADs were characterized with chronoamperometry using ferrocenylmethyl trimethylammonium as the electrochemical probe. Optimized flow rates and injection volumes gave analysis times roughly an order of magnitude faster than those of previously reported flow injection analysis ePADs. To demonstrate applicability, the CF-TPE-ePADs were used to quantify caffeic acid in three different tea samples. The proposed method had a linear range from 10 to 500 μmol L -1 and limits of detection and quantification of 2.5 and 8.3 μmol L -1 , respectively. Our approach is promising for fabricating simple, inexpensive, yet high-performance, flow injection analysis devices using paper substrates and easy-to-make electrodes that do not require external mechanical pumping systems or complicated valves.

Falsified and substandard antibiotics are a growing worldwide problem that leads to increased patient mortality and decreased trust in healthcare, and contributes to antimicrobial resistance. Monitoring falsified antibiotics is difficult because most falsified pharmaceuticals are most commonly found in developing countries, where detecting the active ingredient is difficult due to lack of access to complex instrumentation. Herein, we describe the development and optimization of a microfluidic paper-based analytical device (μPAD) to detect the active ingredient in the most falsified class of antibiotics, β-lactams. The assay is based on enzyme competition, making it the first demonstrated competitive enzyme assay reported in paper-based devices. The assay uses nitrocefin, a chromogenic substrate, to compete with β-lactam antibiotics in a reaction with β-lactamase. A yellow color indicates legitimate drugs, while a color change from yellow to red indicates falsified drugs. In addition to testing for the active ingredient, another section of the device was added to test the sample pH to further verify results and identify common falsified ingredients like aspirin or baking soda. Calibration curves for four different antibiotics, including cefazolin, have been generated making it the first paper-based device capable of detecting both cephalosporin and penicillin antibiotics. The μPAD has also been tested with common falsified ingredients and four antibiotics in tablet or injectable form, demonstrating its potential for in-field falsified antibiotic testing.