Microfluidic Paper-based Analytical Devices Research Articles

Development of a novel three-dimensional microfluidic paper-based analytical device (3D-μPAD) for chlorpyrifos detection using graphene quantum-dot capped gold nanocomposite for colorimetric assay

ABSTRACT This report presents a three-dimensional microfluidic paper-based analytical device (3D-μPAD) with colorimetric assay, for chlorpyrifos organophosphate pesticide detection in vegetable samples. The 3D-µPAD was fabricated by one-step polymer-screen-printing, using rubber latex (RL) waste as a hydrophobic reagent for low-cost and simple manufacture. The 3D-µPAD design comprises two sheets; a testing sheet containing two circular zones, and a sampling sheet in the shape of a dumbbell design. Assay involves the acetylcholinesterase (AChE)-catalysed hydrolysis of an acetylthiocholine (ATCh) substrate to produce thiocholine. Thiocholine causes the aggregation of graphene-quantum-dot capped gold-nanocomposite particles (GQD-AuNPs) to give a purple–blue-coloured solution. Incubation with chlorpyrifos inhibits the hydrolysis reaction, resulting in anti-aggregation of red-coloured GQD-AuNPs. The assay can determine chlorpyrifos by ImageJ detection, over a linear range of 0.001 to 1.0 µg mL−1, with a detection limit of 0.0007 µg mL−1, without sophisticated instrumentation. The developed 3D-µPAD was applied to detect chlorpyrifos in spiked vegetable samples, with per cent recoveries ranging from 93.0% to 104.6%. Our developed device provides good precision (%RSD ranges from 0.3 to 1.6). The calculated relative error comparison with HPLC ranges from 1.0% to 5.2%, indicating a high degree of accuracy. The 3D-µPAD exhibits good sensitivity and selectivity for a low-cost and rapid-screening test for the presence of insecticides, and might be useful for on-site applications.

Fabrication, Flow Control, and Applications of Microfluidic Paper-Based Analytical Devices.

Paper-based microfluidic devices have advanced significantly in recent years as they are affordable, automated with capillary action, portable, and biodegradable diagnostic platforms for a variety of health, environmental, and food quality applications. In terms of commercialization, however, paper-based microfluidics still have to overcome significant challenges to become an authentic point-of-care testing format with the advanced capabilities of analyte purification, multiplex analysis, quantification, and detection with high sensitivity and selectivity. Moreover, fluid flow manipulation for multistep integration, which involves valving and flow velocity control, is also a critical parameter to achieve high-performance devices. Considering these limitations, the aim of this review is to (i) comprehensively analyze the fabrication techniques of microfluidic paper-based analytical devices, (ii) provide a theoretical background and various methods for fluid flow manipulation, and (iii) highlight the recent detection techniques developed for various applications, including their advantages and disadvantages.

Chromium speciation using paper-based analytical devices by direct determination and with electromembrane microextraction

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).

Three-dimensional microfluidic paper-based device for multiplexed colorimetric detection of six metal ions combined with use of a smartphone.

A simple double-layered three-dimensional (3D) microfluidic paper-based analytical device (μPAD) was designed for the simultaneous determination of six metal ions-Fe(III), Ni(II), Cr(VI), Cu(II), Al(III), and Zn(II)-for the first time. The 3D μPAD was composed of two paper layers: a top pretreatment layer and a bottom colorimetric detection layer. The sample solution added to the central sample reservoir of the 3D μPAD could be automatically divided into eight flow pathways and be automatically pretreated while flowing through the pretreatment zones located in the microfluidic channels, and automatically carried out the chromogenic reactions after reaching the detection zones. Random diffusion of the chromogenic reagents was effectively prevented by transport of the pretreated sample solution to the detection zones through 3D microfluidic channels with an L-type circuitous flow route design, resulting in highly increased color uniformity and reproducibility. Combined with use of a flat LED lamp as an upward lighting source and a smartphone as a convenient detector, improved color perception, highly enhanced sensitivity, and an extended detection range were obtained. Finally, the double-layered 3D μPAD was applied to the multiplexed determination of the six metal ions in mixtures and environmental samples with satisfactory results. Detection limits as low as 0.2, 0.3, 0.1, 0.03, 0.08, and 0.04 mg/L for Fe(III), Ni(II), Cr(VI), Cu(II), Al(III), and Zn(II) detection, respectively, were achieved, which are about one order of magnitude lower than obtained with previously reported μPADs for the detection of metal ions. The present 3D μPAD is simple, fast, selective, sensitive, and user-friendly, and holds great application potential for multiplexed on-site analysis.

Paper based immunosensing of ovarian cancer tumor protein CA 125 using novel nano-ink: A new platform for efficient diagnosis of cancer and biomedical analysis using microfluidic paper-based analytical devices (μPAD)

Ovarian cancer is the first and most important cause of malignancy death in women. Mucin 16 or MUC16 protein also known as carcinoma antigen 125 (CA 125) is the most commonly used glycoprotein for early stage diagnosis of ovarian cancer. In this work, a novel paper-based bio-device through hand writing of Ag/RGO (silver nanoparticles/reduced graphene oxide) nano-ink on the flexible paper substrate using pen-on-paper technology was developed. The prepared interface was used to the recognition of CA 125 protein in human biofluid. For this purpose, Ag/rGO nano-ink was synthesized by deposition of Ag nanoparticles onto graphene oxide sheets and the reduction of graphene oxide to rGO simultaneously. Conductivity and resistance of conductive lines were studied after drawing on photographic paper. Subsequently, to prepare a new and unique immuno-device, paper electrode modified by cysteamine caped gold nanoparticles (CysA/Au NPs) using electrochemical techniques. CysA is bonded by sulfur atoms with Au (CysA/Au NPs), and from the amine group with hydroxyl and carboxyl groups of Ag/RGO nano-ink deposited on the surface of paper-based electrodes (CysA/Au NPs/Ag-rGO). Then, anti-CA 125 antibody was immobilized on the electrode surface through Au NPs and CA 125 positively charged amine groups interaction. Atomic force microscopy, Transmission electron microscopy, Field emission scanning electron microscopy, and dynamic light scattering, were performed to identify the engineered immunosensor. Using chronoamperometry technique and under the optimized conditions, the low limit of quantitation (LLOQ) for the proposed immunoassay was recorded as 0.78 U/ml, which this evaluation was performed at highly linear range of 0.78–400 U/ml. The high sensitivity of the electrochemical immunosensor device is indicative of the ability of this immuno-device to detect early stages ovarian cancer.

Isoelectric focusing on microfluidic paper-based chips

Isoelectric focusing (IEF), a powerful technique for protein separation and enrichment, was successfully integrated into microfluidic paper-based analytical devices (μPADs) in this work. The μPADs for isoelectric focusing were fabricated by octadecyltrichlorosilane (OTS) silanization and subsequent region-selective plasma treatment. The system of IEF on μPADs could be easily assembled. And a series of conditions of the system were investigated, including the suitable concentration of ampholyte to create good pH gradient, the effect of polyvinylpyrrolidone (PVP) on electroosmotic flow (EOF) suppression, and focusing voltage applied on the paper channel. After optimization, simultaneous separation and enrichment of protein sample containing myoglobin and cytochrome C was successfully demonstrated. Besides, parallel IEF on multichannels were also achieved for the separation of multiple protein samples on one single chip, and their performance was compared with that of the conventional gel-IEF system. The developed IEF on μPADs exhibits appealing features such as low cost, simplicity, and disposability and are believed to have great application potentials.

Screen-printed microfluidic paper-based analytical device (μPAD) as a barcode sensor for magnesium detection using rubber latex waste as a novel hydrophobic reagent

This work reports the first use of cis-1,4-polyisoprene obtained from rubber latex (RL) waste as the hydrophobic reagent for the fabrication of a microfluidic paper-based analytical device (μPAD), providing a user-friendly means for magnesium detection. The μPAD was fabricated using a screen printing technique and the barcode-like paper sensor was then used for the detection of Mg(II) ions in RL and water samples. Using different types of paper media (paper towel, Whatman No.1 and Whatman No.4), the results indicate that the key factors in optimizing the quality of the fabricated μPAD include the viscosity of cis-1,4-polyisoprene solution which could be adjusted using different solvents and heating temperatures, the mesh screen size, the pore size of the paper substrates, and the dimension of the sample zone. The fabricated μPAD, which showed high chemical resistance, durability and design flexibility, was tested for the detection of Mg(II) ions using the reaction based on complexometric titration with EDTA where Eriochrome Black T was used as an indicator. An Android application “UBU OMg Sensor” was also developed to provide a simple, fast, and accurate means for end-users to interpret results generated by our developed μPAD.

Fabrication of paper-based microfluidic device by recycling foamed plastic and the application for multiplexed measurement of biomarkers.

Microfluidic paper-based analytical devices (μPADs) are emerging as effective analytical platforms for point-of-care assays in resource-limited areas. Simple and cost-effective fabrication method still remains challenging on μPADs. A simple and cost-effective method for fabricating paper-based devices was presented in this work by using of dipping strategy with the recycled polystyrene in chloroform as the hydrophobic reagent. Adhesive tape was employed as mask to transfer the hydrophilic channel pattern to the paper substrate. With the single-sided adhesive tape stuck on the hydrophilic parts of the paper surface, the paper-based device was immersed in chloroform solution with dissolving recycling polystyrene for several seconds. Then the hydrophilic pattern can be achieved and all the other parts on the paper surface were hydrophobic. The adhesive tape was torn off from the hydrophilic parts. The highest contact angle value of 114° of the hydrophobic part was acquired with this simple fabrication method. By using of the sandwich-type immunoreactions and luminol-H2O2p-iodophenol (PIP) chemiluminescence(CL) system, three cancer biomarkers were simultaneously detected in human serum samples on μPADs with the linear range of 0.05-80.0 ng·mL-1 for carcinoembryonic antigen (CEA), 5.0-80.0 ng·mL-1 for alpha-fetal protein (AFP) and 1.0-50.0 ng·mL-1 for prostate-specific antigen (PSA). The fabricating strategy with recycling polystyrene and adhesive tape provides a versatile platform for prototyping of μPADs in both developed and resource constrained region.

Rapid Bacteria Detection at Low Concentrations Using Sequential Immunomagnetic Separation and Paper-Based Isotachophoresis.

Detecting bacteria is important in the fields of human health, environmental monitoring, and food safety. Foodborne pathogens alone are estimated to cause 420 000 deaths annually, with low-income regions affected most. Despite improvements in bacterial detection, fast, disposable, low-cost, sensitive, and user-friendly methods are still needed. Traditional methods for detecting bacteria rely primarily on cell culturing or polymerase chain reaction (PCR), which require highly trained personnel and a central laboratory and take several hours or even days to deliver results. Low-cost methods like lateral flow immunoassays exist but frequently suffer from poor sensitivity and/or lack quantitative results. Here, a rapid method for detecting bacteria at very low concentrations is presented using two sequential preconcentration steps. In the first preconcentration step, the sample is mixed with antibody-modified magnetic particles and free antibodies conjugated to β-galactosidase (β-gal). The target bacteria are isolated and concentrated using immunomagnetic separation. The isolated bacteria are then incubated with chlorophenol red-β-d-galactopyranoside (CPRG), which reacts with β-gal to produce chlorophenol red (CPR) in a bacteria concentration-dependent manner. In the second step, CPR and CPRG are separated and focused using an isotachophoretic microfluidic paper-based analytical device, significantly improving the final detection limit relative to paper-based devices lacking the focusing mechanism. Moreover, CPR and CPRG form two visible color bands that act as test and control bands, respectively, improving assay robustness. The method was tested with E. coli DH5-α and successfully detected concentrations as low as 9.2 CFU/mL in laboratory samples and 920 CFU/mL in apple juice samples in ∼90 min.

Pesticide Residues Identification by Optical Spectrum in the Time-Sequence of Enzyme Inhibitors Performed on Microfluidic Paper-Based Analytical Devices (µPADs).

Pesticides vary in the level of poisonousness, while a conventional rapid test card only provides a general “absence or not” solution, which cannot identify the various genera of pesticides. In order to solve this problem, we proposed a seven-layer paper-based microfluidic chip, integrating the enzyme acetylcholinesterase (AChE) and chromogenic reaction. It enables on-chip pesticide identification via a reflected light intensity spectrum in time-sequence according to the different reaction efficiencies of pesticide molecules and assures the optimum temperature for enzyme activity. After pretreatment of figures of reflected light intensity during the 15 min period, the figures mainly focused on the reflected light variations aroused by the enzyme inhibition assay, and thus, the linear discriminant analysis showed satisfying discrimination of imidacloprid (Y = −1.6525X − 139.7500), phorate (Y = −3.9689X − 483.0526), and avermectin (Y = −2.3617X − 28.3082). The correlation coefficients for these linearity curves were 0.9635, 0.8093, and 0.9094, respectively, with a 95% limit of agreement. Then, the avermectin class chemicals and real-world samples (i.e., lettuce and rice) were tested, which all showed feasible graphic results to distinguish all the chemicals. Therefore, it is feasible to distinguish the three tested kinds of pesticides by the changes in the reflected light spectrum in each min (15 min) via the proposed chip with a high level of automation and integration.

T-shirt ink for one-step screen-printing of hydrophobic barriers for 2D- and 3D-microfluidic paper-based analytical devices

This work presents the use of polyvinyl chloride (PVC) fabric ink, commonly employed for screening t-shirts, as new and versatile material for printing hydrophobic barrier on paper substrate for microfluidic paper-based analytical devices (μPADs). Low-cost, screen-printing apparatus (e.g., screen mesh, squeegee, and printing table) and materials (e.g. PVC ink and solvent) were employed to print the PVC ink solution onto Whatman filter paper No. 4. This provides a one-step strategy to print flow barriers without the need of further processing except evaporation for 3–5 min in a fume hood to remove the solvent. The production of the single layer μPADs is reasonably high with up to 77 devices per screening with 100% success rate. This method produces very narrow fluidic channel 486 ± 14 μm in width and hydrophobic barrier of 642 ± 25 μm thickness. Reproducibility of the production of fluidic channels and zones is satisfactory with RSDs of 2.9% (for 486-μm channel, n = 10), 3.7% (for 2-mm channel, n = 50) and 1.5% (for 6-mm diameter circular zone, n = 80). A design of a 2D-μPAD produced by this method was employed for the colorimetric dual-measurements of thiocyanate and nitrite in saliva. A 3D-μPADs with multiple layers of ink-screened paper was designed and constructed to demonstrate the method's versatility. These 3D-μPADs were designed for gas-liquid separation with in-situ colorimetric detection of ethanol vapor on the μPADs. The 3D-μPADs were applied for direct quantification of ethanol in beverages and highly colored pharmaceutical products. The printed barrier was resistant up to 8% (v/v) ethanol without liquid creeping out of the barrier.

Novel 3D Printed Microfluidic Paper-Based Analytical Device With Integrated Screen-Printed Electrodes for Automated Viscosity Measurements

Various miniaturized viscometers have been developed utilizing several fabrication methods. Among them, microfluidic paper-based analytical devices ( $\mu $ PADs) are becoming popular due to their fabrication ease, cost-effectiveness, and the fact that the flow can be carried out using the embedded capillaries themselves. Mostly, $\mu $ PADs are reported to be fabricated by a solid-ink printer, which has significantly high capital and operational cost. To overcome such drawbacks, a novel rapid prototyping method has been proposed, wherein the formation of the hydrophobic regions was created by polycaprolactone (PCL) filament using a 3-D printer. To leverage this, $\mu $ PAD as a viscometer, velocity, or time between two points with known distances was required, which was carried out by an amperometric approach, established by fabricating the integrated screen-printed electrodes intersecting the microchannel of the $\mu $ PAD. The time measurement was fully automated by a microcontroller, and the relative viscosity was calculated by comparing the time taken by the reference fluid with that of a test fluid to cover a known length. Such integrated, automated, and low-cost paper-based microviscometer was leveraged to measure and analyze the viscosities of various milk variants, which has an accuracy of >92%.

High-resolution temporally resolved chemiluminescence based on double-layered 3D microfluidic paper-based device for multiplexed analysis

In this work, a double-layered three-dimensional (3D) microfluidic paper-based analytical device (μPAD) with high resolution temporally resolved chemiluminescence (CL) emissions were designed for multiplexed CL analysis. The temporally resolved CL emissions were obtained by virtue of the 3D branched microfluidic channel design, which create time delays for luminol transported from one detection zone to another. The peak intensity and peak shape of the temporally resolved CL emissions were quite stable and base-line separated with resolution as high as 21.2–24.4. Then, the fabricated μPAD was applied to multiplexed determination of glucose, lactate, cholesterol, and choline as model analytes. The sample was added to four detection zone modified with CL catalyst cobalt ion and different oxidase by virtue of chitosan. When luminol flowed to μPAD, four temporally resolved CL peaks were successively generated from the cobalt ion catalyzed CL reactions between luminol and generated H2O2 from the specific enzymatic reactions between the oxidase and the analytes. The generated four CL emission peaks in the CL kinetic curve increased in proportion to the concentrations of glucose, lactate, cholesterol, and choline, respectively. Finally, four linear calibration curves were obtained for the detection of glucose (0.01–1.0 mmol/L), lactate (0.02–5.0 mmol/L), cholesterol (0.01–0.4 mmol/L), and choline (0.001–1.0 mmol/L). The detection limits were as low as 8 μmol/L, 15 μmol/L, 6 μmol/L, and 0.07 μmol/L for glucose, lactate, choline, and cholesterol detection, respectively. The present work provides a new strategy for the fabrication of simple and sensitive 3D μPAD with high resolution temporally resolved CL emissions for multiplexed CL analysis, which holds great application potential for point-of-care diagnosis.

Multilayered Microfluidic Paper-Based Devices: Characterization, Modeling, and Perspectives.

Microfluidic paper-based analytical devices (μPADs) are simple but powerful analytical tools that are gaining significant recent attention due to their many advantages over more traditional monitoring tools. These include being inexpensive, portable, pump-free, and having the ability to store reagents. One major limitation of these devices is slow flow rates, which are controlled by capillary action in the hydrophilic pores of cellulosic paper. Recent investigations have advanced the flow rates in μPADs through the generation of a gap or channel between two closely spaced paper sheets. This multilayered format has opened up μPADs to new applications and detection schemes, where large gap sizes (>300 μm) provide at least 169× faster flow rates than single-layer μPADs, but do not conform to established mathematical models for fluid transport in porous materials, such as the classic Lucas-Washburn equation. In the present study, experimental investigations and analytical modeling are applied to elucidate the driving forces behind the rapid flow rates in these devices. We investigate a range of hypotheses for the systems fluid dynamics and establish a theoretical model to predict the flow rate in multilayered μPADs that takes into account viscous dissipation within the paper. Device orientation, sample addition method, and the gap height are found to be critical concerns when modeling the imbibition in multilayered devices.

Inkjet printed microfluidic paper-based analytical device (μPAD) for glucose colorimetric detection in artificial urine.

This article introduces a novel inkjet printing method for the fabrication of a microfluidic paper-based analytical device (μPADs) with improved analytical performance for colorimetric measurements. Firstly, a hydrophobic boundary was created by wax printing on chromatography paper. Then, chitosan (CHI), 3,3',5,5'-Tetramethylbenzidine (TMB) and enzymatic mixture solvent (glucose oxidase (GOx) and horseradish peroxidase (HRP)) were sequentially printed in the sensing zone. Polyethylene glycol (PEG6000) was mixed with the bienzymatic solution to act as an enzyme stabilizer, forming the printable ink. The resulting μPADs exhibited a linear relationship between color intensity and glucose concentration from 0.025mg/ml to 0.5mg/ml. The detectable glucose concentration was in a clinically relevant range from 0.01mg/ml to 4mg/ml. The limit of detection (LOD) was achieved at 0.01mg/ml. After 60-day storage under 4°C, the color intensity at the testing zone retained over 80% of the original intensity. In addition, a smartphone application was developed for in situ colorimetric image processing, and the colorimetric analysis results were compared with those from the use of a scanner followed by processing using ImageJ. Furthermore, the development of this ink printing method also provides a point of care (POC) platform for other substances detection purposes.

A paper-based microfluidic analytical device combined with home-made SPE column for the colorimetric determination of copper(II) ion

Monitoring of copper(II) ion is critical for water safety. Herein, a solid phase extraction (SPE) column filled with home-made sulfonated PS-DVB microspheres was adopted to eliminate interference from other co-existing ions. Meanwhile, 1-(2-pyridylazo)-2-naphthol (PAN) as colorimetric reactant was dispersed in polyvinyl butyral (PVB) and immobilized on the filter paper. The as-fabricated paper-based microfluidic analytical device (μPAD) combined with home-made SPE column was utilized for the selective and quantitative determination of copper(II) ion. The selectivity and sensitivity for copper(II) ion were significantly enhanced by SPE-combined μPAD. The sensor has a noticeable color change in the presence of copper(II) ion. The limit of detection was as low as 0.340 μM, which is much less than the drinking water guidelines in many countries and organizations. This technology is convenient and does not need any expensive instrumentation and professional skills. It is easily used to achieve rapid and on-site assessment of copper(II) ion in real-world situations.

Developing a Mickey-Mouse-Designed Microfluidic Paper-Based Analytical Device (μPAD) to Determine The Antioxidant Activity of Green Tea

The determination of the antioxidant activity in green tea using FRAP essays developed in a Microfluidic Paper-Based Analytical Device (μPADs) was observed. μPAD was prepared on the chromatographic paper with a suitable pattern and then printed using a solid ink printer. The solid printing was intended to obtain a hydrophobic barrier and a hydrophilic channel on the chromatographic paper. A preliminary study was done to determine the optimum time and temperature used on the penetration of obtaining the hydrophobic barrier to avoid leakage in the channel. Optimization of temperature and time was calculated by the average velocity, the optimum condition was obtained at 120 °C for 90 seconds with an average speed of 0.1 mm.s−1. The green tea samples were prepared by extracting its active compound using demineralized water at 25 °C and 90 °C with 2 hours of immersion time. For the measurement of antioxidant activities, the analysis was carried out by placing 0.5 μl in the detection zone and 5 μl samples into the sample zone μPAD. Then, the color reaction product, which propagates from the sample zone to the detection zone, is processed by Image J software to measure color intensity in CMYK mode. Extracts of green tea samples at 25 °C and 90 °C has a significant difference in antioxidant activity. These results indicate that the method developed in this work can be used as an alternative method for analyzing antioxidant activity in green tea extract. The results of the average amount of antioxidant activity in green tea samples were shown with Fe2+ concentration. High-speed detection, low cost, high accuracy, and ease of use can be attributed to the advantages of our μPAD method.

Microfluidic Paper based Analytical Device (µpads) for Analysis of Benzoat Acid in Packaged Beverages

Microfluidic Paper-based Analytical Devices (μPADs) are a simple analytical platform that satisfy combination of low cost, portability, and ease-to-use with selectivity, sensitivity, and accuracy. In this experiment, the μPADs devices are prepared using a chromatographic paper and designed at appropriate pattern prior to printing by mean of a solid ink (wax) printer to produce hydrophobic barriers and hydrophilic channels. Then, the μPADs are heated at 120°C for 3 min to allow a wax penetrates in the both side of a paper. The resulted μPADs are used for the detection of benzoic acid through alkalimetry principle. For this purpose, the devices are implanted with 5M NaOH in the detection zone and a 2% phenolphthalein indicator in the sample zone before dried in open air. When the devices have dried, the μPADs are ready to be used to detect benzoic acid. To avoid color interferences, the samples are distilled before introduced to the sample zone for quantitative detection. The result soft the color changes can be immediately seen by naked eye, and their intensities are quantified by the Image J software. Excellent linearity is achieved in the range of benzoic acid concentration of 20-120 ppm with the linear equation of Y = 0.0102 x + 0.0077 and the correlation coefficient (R2) of 0.9999. The optimized μPADs devices are successfully applied to the quantitative analysis of benzoic acid in the 5 kinds of commercially packaged beverages. There is no significantly different of the analytical results obtained by the µPADs in comparison to the spectrophotometric method. Overall, the results obtained in this study indicate that the μPADs devices are a reliable tool for high throughout and on-site determination of benzoic acid content in commercial drinks.

Simultaneous Determination Of BUN-Creatinine as Kidney Function Biomarkers in Blood using a Microfluidic Paper-based Analytical Devices

In this paper, we describe a Microfluidic Paper-based Analytical Devices (μPADs) for the simultaneous quantification of two important biomarkers of kidney function in blood. This paper provides a simple, disposable, portable, and inexpensive colorimetric method for the quantification of Blood Urea Nitrogen (BUN) and serum creatinine (sCRE). BUN detection is based on the Berthelot reaction, in which urea is converted to ammonium by the use of urease, ammonium ion which then reacts with a mixture of salicylate, sodium nitroprusside, and hypochlorite to yield a blue-green chromophore. sCRE detection is based on the Jaffe reaction, in which sCRE under alkaline conditions react with picrate ions forming an orange complex. The intensity of the color formed is proportional to the BUN & sCRE concentration in the sample. Various experimental parameters were optimized to achieve the best performance of the μPADs. There were no significant differences between the results obtained using the μPADs and a comparative method.

Microfluidic Paper-based Analytical Devices (µPADs) For Analysis Lead Using Naked Eye and Colorimetric Detections

This work describes comparation two designs of a microfluidic paper-based analytical devices (µPADs) using a wax printing for fabrication of hydrophobic zones on the chromatographic paper and their application to the detection of lead in waste samples. Two different designs used in this research were distance-based method and analysis image method. Sodium rhodizonate in tartrate buffer solution (pH 2.8) solution are used as the colorimetric reagents for the direct naked eyed detection of lead(II) on µPADs. The detection of lead(II) concentration is conducted by measuring the distanceof a colored reaction product which propagate in the detection zone (design 1) and the change in color intensity on the µPADs is also used for the lead detection using ImageJ software to determine the RGB value (design 2). Related to both methods can be used to determine the concentration of lead(II) with a comparable coefficient of determination, but based on the analysis method, design one is a design that is very simple, easy, fast, inexpensive and only uses the naked eye for detection.