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Over the last few years, the SARS-CoV-2 pandemic has made the need for rapid, affordable diagnostics more compelling than ever. While traditional laboratory diagnostics like PCR and well-plate ELISA are sensitive and specific, they can be costly and take hours to complete. Diagnostic tests that can be used at the point-of-care or at home, like lateral flow assays (LFAs) are a simple, rapid alternative, but many commercially available LFAs have been criticized for their lack of sensitivity compared to laboratory methods like well-plate ELISAs. The Capillary-Driven Immunoassay (CaDI) device described in this work uses microfluidic channels and capillary action to passively automate the steps of a traditional well-plate ELISA for visual read out. This work builds on prior capillary-flow devices by further simplifying operation and use of colorimetric detection. Upon adding sample, an enzyme-conjugated secondary antibody, wash steps, and substrate are sequentially delivered to test and control lines on a nitrocellulose strip generating a colorimetric response. The end user can visually detect SARS-CoV-2 antigen in 15-20 min by naked eye, or results can be quantified using a smartphone and software such as ImageJ. An analytical detection limit of 83 PFU/mL for SARS-CoV-2 was determined for virus in buffer, and 222 PFU/mL for virus spiked into nasal swabs using image analysis, similar to the LODs determined by traditional well-plate ELISA. Additionally, a visual detection limit of 100 PFU/mL was determined in contrived nasal swab samples by polling 20 untrained end-users. While the CaDI device was used for detecting clinically relevant levels of SARS-CoV-2 in this study, the CaDI device can be easily adapted to other immunoassay applications by changing the reagents and antibodies., Competing Interests: Declaration of competing interest 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., (Copyright © 2023 Elsevier B.V. All rights reserved.)

At home electrolyte analysis is of growing interest due to the importance for early diagnosis for various diseases. This work presents the first device that integrates a potentiometric ion-selective electrode (ISE) with distance-based colorimetric detection for the simultaneous analysis of K + and Cl - ions at the point of care (POC). This hybrid sensing device was designed in a two-dimensional configuration using a plastic transparency sheet containing a stencil printed K + -ISE and a paper substrate for distance-based colorimetric detection of Cl - . K + quantification was performed using a low-cost (<$25) lab-made Wi-Fi supported potentiometer with a custom smartphone application while Cl - ions were quantified with an instrument-free distance-based paper analytical device (dPAD). The total analysis time is 5 min once the sample is added. The K + -ISE showed a sensitivity of 55.89 ± 2.52 mV/decade from 0.1 to 100 mM with a limit of detection (LOD) of 0.01 mM. The linear working range for Cl - detection was 0.5-50 mM whereas the LOD was 0.16 ± 0.05 mM. The proof-of-concept application of the sensing hybrid device was demonstrated in human urine and artificial sweat samples containing K + and Cl - ions at physiologically relevant ranges. The recoveries were found to be 88-108% for K + and 90-104% for Cl - , showing the potential use of the proposed device for low-cost routine analysis of biological samples at POC., Competing Interests: Declaration of competing interest 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., (Copyright © 2022 Elsevier B.V. All rights reserved.)

This study reports a new electrochemical method for tryptamine determination using a paper-based microfluidic device and a thermoplastic electrode (TPE) as an amperometric detector. Tryptamine (Tryp) is a biogenic amine present in drinks and foods. Even though this compound has some beneficial effects on human health, the ingestion of foods with high concentrations of Tryp may be detrimental, which justifies the need for monitoring the Tryp levels. The TPEs were made from 50% carbon black and 50% polycaprolactone and characterized by cyclic voltammetry, demonstrating enhancement in the analytical response compared to other carbon composites. TPEs also showed a better antifouling effect for Tryp compared to conventional glassy carbon electrodes. Once characterized, the electrodes were incorporated into the microfluidic device to determine Tryp in water and cheese samples using amperometry. A linear range was achieved from 10 to 75 μmol L -1 with limits of detection and quantification of 3.2 and 10.5 μmol L -1 , respectively. Therefore, this work shows promising findings of the electrochemical determination of Tryp, bringing valuable results regarding the electrochemical properties of thermoplastic composites., Competing Interests: Declaration of competing interest 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., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Microfluidics has revolutionized the fields of bioanalytical chemistry, cellular biology, and molecular biology. Advancements in microfluidic technologies, however, are often limited by labor, time, and resource-intensive fabrication methods, most commonly a form of photolithography. The advent of 3D printing has helped researchers fabricate proof-of-concept microfluidics more rapidly and at lower costs but suffers from poor resolution and tedious post-processing to remove uncured resin from enclosed channels. Additionally, custom resins and printers are often needed to create entirely enclosed channels, which increases cost and complexity of fabrication. In this work we demonstrate the ability to create microfluidic devices by covalently sealing 3D-printed parts with open-faced channels to polydimethylsiloxane (PDMS). Open-faced channels are easier to print than fully enclosed channels and can be printed using an inexpensive and commercially available stereolithography 3D printer and resin. The 3D-printed parts are sealed to PDMS, a common substrate used in traditional microfluidic fabrication, using two different techniques. The first involves coating the part with a commercially available silicone spray before sealing to PDMS via plasma treatment. In the second technique, the cured methacrylate resin is silanized with (3-Aminopropyl)triethoxysilane (APTES) before binding to PDMS with plasma treatment. Both methods create a strong seal between the two substrates, which is demonstrated with several types of microfluidic devices including droplet and gradient generators., Competing Interests: Declaration of competing interest 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., (Copyright © 2020. Published by Elsevier B.V.)

The use of paper microfluidics to perform chemical measurements for various analytical applications has gained interest over the last decade. One of the growing applications of these platforms is for the qualitative and quantitative determination of drugs. The low cost and self-pumping ability of paper microfluidics are attractive for developing analytical tools capable of on-site drug screening. This review aims to present the unique features of microfluidic paper-based analytical devices (μPADs) that offer advantages to pharmaceutical analysis and evaluate the state-of-the-art technologies and applications of the platform for drug analysis in research and real-world settings. The current challenges and potential future directions of the field are also highlighted., Competing Interests: Declaration of competing interest 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., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Cadmium (Cd) and lead (Pb) pollution is a significant environmental and human health concern, and methods to detect Cd and Pb on site are valuable. Stencil-printed carbon electrodes (SPCEs) are an attractive electrode material for point-of-care (POC) applications due to their low cost, ease of fabrication, disposability and portability. At present, SPCEs are exclusively formulated from graphitic carbon powder and conductive carbon ink. However, graphitic carbon SPCEs are not ideal for heavy metal sensing due to the heterogeneity of graphitic SPCE surfaces. Moreover, SPCEs typically require extensive modification to provide desirable detection limits and sensitivity at the POC, significantly increasing cost and complexity of analysis. While there are many examples of chemically modified SPCEs, the bulk SPCE composition has not been studied for heavy metal detection. Here, a glassy carbon microparticle stencil printed electrode (GC-SPE) was developed. The GC-SPEs were first characterized with SEM and cyclic voltammetry and then optimized for Cd and Pb detection with an in situ Bi-film plated. The GC-SPEs require no chemical modification or pretreatment significantly decreasing the cost and complexity of fabrication. The detection limits for Cd and Pb were estimated to be 0.46 μg L -1 and 0.55 μg L -1 , respectively, which are below EPA limits for drinking water (5 μg L -1  Cd and 10 μg L -1  Pb) [1]. The reported GC-SPEs are advantageous with their low cost, ease of fabrication and use, and attractive performance. The GC-SPEs can be used for low-level metal detection at the POC as shown in the report herein., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: CSH is a cofounder and equity holder in Access Sensor Technologies., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Herein we report the first two distance-based microfluidic paper-based analytical devices (μPADs) using fluorescence to quantify aluminum. In addition to their read-by-eye quantification, the devices are simple to fabricate, require no sample pretreatment or preconcentration, and have a shelf life of >5 months. The first device is designed in a "chemometer" format where the length of a fluorescent band linearly responds to an Al(III) concentration. The second device uses a radial design where the fluorescent diameter also linearly responds to an Al(III) concentration. The chemometer device has a detection limit of 2.5 ppm (100 μM) and a linear range from 2 to 54 ppm Al(III) (100 μM-1 mM), R 2  = 0.989). The radial device has a detection limit of 0.9 ppm (33 μM) and a linear range from 2 to 24 ppm Al(III) (100-900 μM, R 2  = 0.968). The utility of the μPADs were successfully demonstrated by measuring Al(III) in two water effluent samples from the Gold King Mine near Silverton, CO., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: A portion of this work was supported by Access Sensor Technologies through a grant from the National Science Foundation. CSH has an ownership stake in Access Sensor Technologies., (Copyright © 2019. Published by Elsevier B.V.)

The simultaneous detection of multiple analytes from a single sample is a critical tool for the analysis of real world samples. However, this is challenging to accomplish in the field by current electroanalytical techniques, where tuning assay conditions towards a target analyte often results in poor selectivity and sensitivity for other species in the mixture. In this work, an electrochemical paper-based analytical device (ePAD) capable of performing simultaneous electrochemical experiments in different solution conditions on a single sample was developed for the first time. We refer to the system as a Janus-ePAD after the two-faced Greek god because of the ability of the device to perform electrochemistry on the same sample under differing solution conditions at the same time with a single potentiostat. In a Janus-ePAD, a sample wicks down two channels from a single inlet towards two discreet reagent zones that adjust solution conditions, such as pH, before flow termination in two electrochemical detection zones. These zones feature independent working electrodes and shared reference and counter electrodes, facilitating simultaneous detection of multiple species at each species' optimal solution condition. The device utility and applicability are demonstrated through the simultaneous detection of two biologically relevant species (norepinephrine and serotonin) and a common enzymatic assay product (p-aminophenol) at two different solution pH conditions. Janus-ePADs show great promise as an inexpensive and broadly applicable platform which can reduce the complexity and/or number of steps required in multiplexed analysis, while also operating under the optimized conditions of each species present in a mixture., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A label-free electrochemical DNA sensor based on pyrrolidinyl peptide nucleic acid (acpcPNA)-immobilized on a paper-based analytical device (PAD) was developed. Unlike previous PNA-based electrochemical PAD (ePAD) sensors where the capture element was placed directly on the electrode, acpcPNA was covalently immobilized onto partially oxidized cellulose paper allowing regeneration by simple PAD replacement. As an example application, a sensor probe was designed for Mycobacterium tuberculosis (MTB) detection. The ePAD DNA sensor was used to determine a synthetic 15-base oligonucleotide of MTB by measuring the fractional change in the charge transfer resistance (R ct ) obtained from electrochemical impedance spectroscopy (EIS). The R ct of [Fe(CN) 6 ] 3-/4- before and after hybridization with the target DNA could be clearly distinguished. Cyclic voltammetry (CV) was used to verify the EIS results, and showed an increase in peak potential splitting in a similar stepwise manner for each immobilization step. Under optimal conditions, a linear calibration curve in the range of 2-200 nM and the limit of detection 1.24 nM were measured. The acpcPNA probe exhibited very high selectivity for complementary oligonucleotides over single-base-mismatch, two-base-mismatch and non-complementary DNA targets due to the conformationally constrained structure of the acpcPNA. Moreover, the ePAD DNA sensor platform was successfully applied to detect PCR-amplified MTB DNA extracted from clinical samples. The proposed paper-based electrochemical DNA sensor has potential to be an alternative device for low-cost, simple, label-free, sensitive and selective DNA sensor., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Coupling infrared (IR) spectroscopy to microfluidic devices provides a powerful tool for characterizing complex chemical and biochemical reactions. Examples of microfluidic devices coupled with infrared spectroscopy have been limited, however, largely due to the difficulties associated with fabricating systems in common infrared transparent materials like CaF 2 . Recent reports have shown that polydimethylsiloxane (PDMS) can be used as an IR transparent substrate when fabricated with thin layers. The use of soft lithography with PDMS expands the library of possible designs that can be achieved for IR measurements in microfluidics. In initial reports with thin PDMS, the target analytes were small molecules; however, IR spectroscopy offers a powerful tool to study protein structure and reactions. Here, a PDMS microfluidic device compatible with IR spectroscopy was fabricated by means of spin-coating of PDMS pre-polymer to obtain thin PDMS microfluidic features. The device was comprised of only PDMS and IR absorption of PDMS was significantly minimized due to the thickness (∼40 μm) of the PDMS layer. The use of thin PDMS allowed for measuring the amide I and II vibrational bands of proteins that have been difficult to measure in other microfluidic devices. To demonstrate the power of the system, the microfluidic device was successfully used to measure the enzyme kinetics as one class of important biochemical reactions with broad use in a variety of fields from medicine to biotechnology. As a model, the reaction of glucose oxidase with glucose was tracked by following the formation of gluconic acid. Michaelis-Menten kinetics from the device were compared with bulk solution measurements and found to be in good agreement., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The field of microfluidic paper-based analytical devices (μPADs) is most notably characterized by portable and low-cost analysis; however, struggles to achieve the high sensitivity and low detection limits needs required for many environmental applications hinder widespread adoption of this technology. Loss of analyte to the device material represents an important problem impacting sensitivity. Critically, we found that at least 50% of a Ni(II) sample is lost when being transported down a 30 mm paper channel that is representative of structures commonly found in μPADs. In this work, we report simple strategies such as adding a waste zone, enlarging the detection zone, and using an elution step to increase device performance. A μPAD combining the best performing functionalities led to a 78% increase in maximum signal and a 28% increase in sensitivity when transporting Ni(II) samples. Using the optimized μPAD also led to a 94% increase in maximum signal for Mn(II) samples showing these modifications can be applied more generally., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In this work, we report a simple and yet efficient stencil-printed electrochemical platform that can be integrated into the caps of sample containers and thus, allows in-field quantification of Cd(II) and Pb(II) in river water samples. The device exploits the low-cost features of carbon (as electrode material) and paper/polyester transparency sheets (as substrate). Electrochemical analysis of the working electrodes prepared on different substrates (polyester transparency sheets, chromatographic, tracing and office papers) with hexaammineruthenium(III) showed that their electroactive area and electron transfer kinetics are highly affected by the porosity of the material. Electrodes prepared on transparency substrates showed the best electroanalytical performance for the simultaneous determination of Cd(II) and Pb(II) by square-wave anodic stripping voltammetry. Interestingly, the temperature and time at which the carbon ink was cured had significant effect on the electrochemical response, especially the capacitive current. The amount of Cd and Pb on the electrode surface can be increased about 20% by in situ electrodeposition of bismuth. The electrochemical platform showed a linear range comprised between 1 and 200 μg/L for both metals, sensitivity of analysis of 0.22 and 0.087 μA/ppb and limits of detection of 0.2 and 0.3 μg/L for Cd(II) and Pb(II), respectively. The analysis of river water samples was done directly in the container where the sample was collected, which simplifies the procedure and approaches field analysis. The developed point-of-need detection system allowed simultaneous determination of Cd(II) and Pb(II) in those samples using the standard addition method with precise and accurate results., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Simple methods have been developed for fabricating microfluidic paper-based analytical devices (μPADs) but few of these devices can be used with organic solvents and/or aqueous solutions containing surfactants. This study describes a simple fabrication strategy for μPADs that uses readily available scholar glue to create the hydrophobic flow barriers that are resistant to surfactants and organic solvents. Microfluidic structures were defined by magnetic masks designed with either neodymium magnets or magnetic sheets to define the patter, and structures were created by spraying an aqueous solution of glue on the paper surface. The glue-coated paper was then exposed to UV/Vis light for cross-linking to maximize chemical resistance. Examples of microzone arrays and microfluidic devices are demonstrated. μPADs fabricated with scholar glue retained their barriers when used with surfactants, organic solvents, and strong/weak acids and bases unlike common wax-printed barriers. Paper microzones and microfluidic devices were successfully used for colorimetric assays of clinically relevant analytes commonly detected in urinalysis to demonstrate the low background of the barrier material and generally applicability to sensing. The proposed fabrication method is attractive for both its ability to be used with diverse chemistries and the low cost and simplicity of the materials and process., (Copyright © 2017 Elsevier B.V. All rights reserved.)

A novel paper-based electrochemical biosensor was developed using an anthraquinone-labeled pyrrolidinyl peptide nucleic acid (acpcPNA) probe (AQ-PNA) and graphene-polyaniline (G-PANI) modified electrode to detect human papillomavirus (HPV). An inkjet printing technique was employed to prepare the paper-based G-PANI-modified working electrode. The AQ-PNA probe baring a negatively charged amino acid at the N-terminus was immobilized onto the electrode surface through electrostatic attraction. Electrochemical impedance spectroscopy (EIS) was used to verify the AQ-PNA immobilization. The paper-based electrochemical DNA biosensor was used to detect a synthetic 14-base oligonucleotide target with a sequence corresponding to human papillomavirus (HPV) type 16 DNA by measuring the electrochemical signal response of the AQ label using square-wave voltammetry before and after hybridization. It was determined that the current signal significantly decreased after the addition of target DNA. This phenomenon is explained by the rigidity of PNA-DNA duplexes, which obstructs the accessibility of electron transfer from the AQ label to the electrode surface. Under optimal conditions, the detection limit of HPV type 16 DNA was found to be 2.3 nM with a linear range of 10-200 nM. The performance of this biosensor on real DNA samples was tested with the detection of PCR-amplified DNA samples from the SiHa cell line. The new method employs an inexpensive and disposable device, which easily incinerated after use and is promising for the screening and monitoring of the amount of HPV-DNA type 16 to identify the primary stages of cervical cancer., (Copyright © 2016 Elsevier B.V. All rights reserved.)

We report herein the first development of graphene-polyaniline modified carbon paste electrode (G-PANI/CPE) coupled with droplet-based microfluidic sensor for high-throughput detection of 4-aminophenol (4-AP) in pharmaceutical paracetamol (PA) formulations. A simple T-junction microfluidic platform using an oil flow rate of 1.8 μL/min and an aqueous flow rate of 0.8 μL/min was used to produce aqueous testing microdroplets continuously. The microchannel was designed to extend the aqueous droplet to cover all 3 electrodes, allowing for electrochemical measurements in a single droplet. Parameters including flow rate, water fraction, and applied detection potential (Edet) were investigated to obtain optimal conditions. Using G-PANI/CPE significantly increased the current response for both cyclic voltammetric detections of ferri/ferrocyanide [Fe(CN)6](3-/4-) (10 times) and 4-AP (2 times), compared to an unmodified electrode. Using the optimized conditions in the droplet system, 4-AP in the presence of PA was selectively determined. The linear range of 4-AP was 50-500 μM (R(2) = 0.99), limit of detection (LOD, S/N = 3) was 15.68 μM, and limit of quantification (LOQ, S/N = 10) was 52.28 μM. Finally, the system was used to determine 4-AP spiked in commercial PA liquid samples and the amounts of 4-AP were found in good agreement with those obtained from the conventional capillary zone electrophoresis/UV-Visible spectrophotometry (CZE/UV-Vis). The proposed microfluidic device could be employed for a high-throughput screening (at least 60 samples h(-1)) of pharmaceutical purity requiring low sample and reagent consumption., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Measurement variance is a crucial aspect of quantitative chemical analysis. Variance directly affects important analytical figures of merit, including detection limit, quantitation limit, and confidence intervals. Most reported analyses for emerging analytical techniques implicitly assume constant variance (homoskedasticity) by using unweighted regression calibrations. Despite the assumption of constant variance, it is known that most instruments exhibit heteroskedasticity, where variance changes with signal intensity. Ignoring nonconstant variance results in suboptimal calibrations, invalid uncertainty estimates, and incorrect detection limits. Three techniques where homoskedasticity is often assumed were covered in this work to evaluate if heteroskedasticity had a significant quantitative impact-naked-eye, distance-based detection using paper-based analytical devices (PADs), cathodic stripping voltammetry (CSV) with disposable carbon-ink electrode devices, and microchip electrophoresis (MCE) with conductivity detection. Despite these techniques representing a wide range of chemistries and precision, heteroskedastic behavior was confirmed for each. The general variance forms were analyzed, and recommendations for accounting for nonconstant variance discussed. Monte Carlo simulations of instrument responses were performed to quantify the benefits of weighted regression, and the sensitivity to uncertainty in the variance function was tested. Results show that heteroskedasticity should be considered during development of new techniques; even moderate uncertainty (30%) in the variance function still results in weighted regression outperforming unweighted regressions. We recommend utilizing the power model of variance because it is easy to apply, requires little additional experimentation, and produces higher-precision results and more reliable uncertainty estimates than assuming homoskedasticity., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Microwire electrodes are presented as an alternative to screen-printed electrodes for detection in electrochemical paper-based analytical devices (ePADs). Compared to carbon ink electrodes, microwire electrodes offer lower resistance and a significant increase in current density relative to carbon ink electrodes. Various microwire compositions and diameters, including 30 μm Pt, 25 μm Au, 18 μm Pt with 8% W, and 15 μm Pt with 20% Ir, were tested and compared to theoretically predicted behavior. The measured current in static solution was below predicted levels for cylindrical microelectrodes but greater than levels predicted for hemi-cylindrical electrodes most likely as a result of the proximity of the electrode to the paper surface. Furthermore, the current response was indicative of semi-thin layer behavior, likely due to the confined solution volume in the paper. After electrode characterization, a device was developed for the non-enzymatic detection of glucose, fructose, and sucrose using a Cu electrode in alkaline solution. The limits of detection for glucose, fructose, and sucrose were 270 nM, 340 nM, and 430 nM, respectively, which are significantly below sugar concentrations found in sweetened beverages or glucose levels in serum., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This work describes the development of an electrochemical sensor for simultaneous detection of Zn(II), Cd(II), and Pb(II) using a graphene-polyaniline (G/PANI) nanocomposite electrode prepared by reverse-phase polymerization in the presence of polyvinylpyrrolidone (PVP). Two substrate materials (plastic film and filter paper) and two nanocomposite deposition methods (drop-casting and electrospraying) were investigated. Square-wave anodic stripping voltammetry currents were higher for plastic vs. paper substrates. Performance of the G/PANI nanocomposites was characterized by scanning electron microscopy (SEM) and cyclic voltammetry. The G/PANI-modified electrode exhibited high electrochemical conductivity, producing a three-fold increase in anodic peak current (vs. the unmodified electrode). The G/PANI-modified electrode also showed evidence of increased surface area under SEM. Square-wave anodic stripping voltammetry was used to measure Zn(II), Cd(II), and Pb(II) in the presence of Bi(III). A linear working range of 1-300 μg L(-1) was established between anodic current and metal ion concentration with detection limits (S/N=3) of 1.0 μg L(-1) for Zn(II), and 0.1 μg L(-1) for both Cd(II) and Pb(II). The G/PANI-modified electrode allowed selective determination of the target metals in the presence of common metal interferences including Mn(II), Cu(II), Fe(III), Fe(II), Co(III), and Ni(II). Repeat assays on the same device demonstrated good reproducibility (%RSD<11) over 10 serial runs. Finally, this system was utilized for determining Zn(II), Cd(II), and Pb(II) in human serum using the standard addition method., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Occupational exposure to Cr is concerning because of its myriad of health effects. Assessing chromium exposure is also cost and resource intensive because the analysis typically uses sophisticated instrumental techniques like inductively coupled plasma-mass spectrometry (ICP-MS). Here, we report a novel, simple, inexpensive microfluidic paper-based analytical device (μPAD) for measuring total Cr in airborne particulate matter. In the μPAD, tetravalent cerium (Ce(IV)) was used in a pretreatment zone to oxidize all soluble Cr to Cr(VI). After elution to the detection zone, Cr(VI) reacts with 1,5-diphenylcarbazide (1,5-DPC) forming 1,5-diphenylcarbazone (DPCO) and Cr(III). The resulting Cr(III) forms a distinct purple colored complex with the DPCO. As proof-of-principle, particulate matter (PM) collected on a sample filter was analyzed with the μPAD to quantify the mass of total Cr. A log-linear working range (0.23-3.75 μg; r(2)=0.998) between Cr and color intensity was obtained with a detection limit of 0.12 μg. For validation, a certified reference containing multiple competing metals was analyzed. Quantitative agreement was obtained between known Cr levels in the sample and the Cr measured using the μPAD., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Electrochemical paper-based analytical devices (ePADs) with integrated plasma isolation for determination of glucose from whole blood samples have been developed. A dumbbell shaped ePAD containing two blood separation zones (VF2 membranes) with a middle detection zone was fabricated using the wax dipping method. The dumbbell shaped device was designed to separate plasma while generating homogeneous flow to the middle detection zone of the ePAD. The proposed ePADs work with whole blood samples with 24-60% hematocrit without dilution, and the plasma was completely separated within 4 min. Glucose in isolated plasma separated was detected using glucose oxidase immobilized on the middle of the paper device. The hydrogen peroxide generated from the reaction between glucose and the enzyme pass through to a Prussian blue modified screen printed electrode (PB-SPEs). The currents measured using chronoamperometry at the optimal detection potential for H2O2 (-0.1 V versus Ag/AgCl reference electrode) were proportional to glucose concentrations in the whole blood. The linear range for glucose assay was in the range 0-33.1 mM (r(2)=0.987). The coefficients of variation (CVs) of currents were 6.5%, 9.0% and 8.0% when assay whole blood sample containing glucose concentration at 3.4, 6.3, and 15.6mM, respectively. Because each sample displayed intra-individual variation of electrochemical signal, glucose assay in whole blood samples were measured using the standard addition method. Results demonstrate that the ePAD glucose assay was not significantly different from the spectrophotometric method (p=0.376, paired sample t-test, n=10)., (Copyright © 2013 Elsevier B.V. All rights reserved.)

We report the development of an electrochemical paper-based analytical device (ePAD) for the selective determination of dopamine (DA) in model serum sample. The ePAD device consists of three layers. In the top layer, SU-8 photoresist defines a hydrophilic sample application spot on the filter paper. The middle layer was made from transparency film and contained two holes, one for sample preconcentration and the other for the surfactant to allow transfer to the third layer. A screen-printed carbon electrode formed the bottom layer and was used for electrochemical measurements. In the absence of the anionic surfactant, sodium dodecyl sulfate (SDS), the oxidation peaks of DA, ascorbic acid (AA) and uric acid (UA) overlapped. With the addition of SDS, the DA oxidation peak shifted to more negative values and was clearly distinguishable from AA and UA. The oxidation potential shift was presumably due to preferential electrostatic interactions between the cationic DA and the anionic SDS. Indeed, whilst the SDS-modified paper improved the DA current five-fold, the non-ionic Tween-20 and cationic tetradecyltrimethylammonium bromide surfactants had no effect or reduced the current, respectively. Furthermore, only the SDS-modified paper showed the selective shift in oxidation potential for DA. DA determination was carried out using square-wave voltammetry between -0.2 and 0.8 V vs. Ag/AgCl, and this ePAD was able to detect DA over a linear range of 1-100 μM with a detection limit (S/N=3) of 0.37 μM. The ePAD seems suitable as a low cost, easy-to-use, portable device for the selective quantitation of DA in human serum samples., (Copyright © 2012 Elsevier B.V. All rights reserved.)

We report here the use of multiple indicators for a single analyte for paper-based microfluidic devices (microPAD) in an effort to improve the ability to visually discriminate between analyte concentrations. In existing microPADs, a single dye system is used for the measurement of a single analyte. In our approach, devices are designed to simultaneously quantify analytes using multiple indicators for each analyte improving the accuracy of the assay. The use of multiple indicators for a single analyte allows for different indicator colors to be generated at different analyte concentration ranges as well as increasing the ability to better visually discriminate colors. The principle of our devices is based on the oxidation of indicators by hydrogen peroxide produced by oxidase enzymes specific for each analyte. Each indicator reacts at different peroxide concentrations and therefore analyte concentrations, giving an extended range of operation. To demonstrate the utility of our approach, the mixture of 4-aminoantipyrine and 3,5-dichloro-2-hydroxy-benzenesulfonic acid, o-dianisidine dihydrochloride, potassium iodide, acid black, and acid yellow were chosen as the indicators for simultaneous semi-quantitative measurement of glucose, lactate, and uric acid on a microPAD. Our approach was successfully applied to quantify glucose (0.5-20 mM), lactate (1-25 mM), and uric acid (0.1-7 mM) in clinically relevant ranges. The determination of glucose, lactate, and uric acid in control serum and urine samples was also performed to demonstrate the applicability of this device for biological sample analysis. Finally results for the multi-indicator and single indicator system were compared using untrained readers to demonstrate the improvements in accuracy achieved with the new system., (2010 Elsevier B.V. All rights reserved.)

Metabolomics is an emerging area of research focused on measuring small molecules in biological samples. There are a number of different types of metabolomics, ranging from global profiling of all metabolites in a single sample to measurement of a selected group of analytes. Microfluidics and related technologies have been used in this research area with good success. The aim of this review article is to summarize the use of microfluidics in metabolomics. Direct application of microfluidics to the determination of small molecules is covered first. Next, important sample preparation methods developed for microfluidics and applicable to metabolomics are covered. Finally, a summary of metabolomic work as it relates to analysis of cellular events using microfluidics is covered.

Immunoassays represent a core workhorse methodology for many applications ranging from clinical diagnostics to environmental monitoring. In traditional formats such as the enzyme linked immunosorbent assay (ELISA), analytes are measured singly or in small sets. As more biomarkers are identified for disease states, there is a need to develop methods that can measure multiple markers simultaneously. Immunoaffinity arrays are one such chemistry that can achieve multi-marker screening. Most arrays are performed in either competitive or non-competitive formats, where the former are used predominantly for small molecules and the later for macromolecules. To date, ELISA and immunoaffinity array methods have relied exclusively on one of these formats and not the other. Here an immunoaffinity array method capable of performing simultaneous competitive and non-competitive analysis generated using micromosaic immunoassay techniques is introduced for the analysis of metabolites and proteins. In this report, three markers of oxidative stress were used as a model system. The method described here demonstrates the simultaneous analysis of 3-nitrotyrosine, by indirect competitive immunoassay while the enzymes catalase and superoxide dismutase are analyzed by non-competitive sandwich immunoassay. The method requires less than 1 microL sample and 45 min for completion. Logistic curve fits and LOD (limits of detection) statistical analysis of the binding results are presented and show good agreement with published data for these antibody-antigen systems.