Paper-based Analytical Devices Research Articles

Paper-based microfluidics: What can we expect?

Paper-based microfluidics: What can we expect?

Paper-based electrochemical platform modified with graphene nanoribbons: A new and affordable approach for analysis of 5-hydroxy-l-tryptophan

Here, a new and inexpensive electrochemical paper-based analytical device (ePAD) containing a pencil-drawn three-electrode setup, in which the working electrode was modified with graphene nanoribbons (GNRs), is presented for the first time. Under optimal conditions, the GNRs-ePAD provided higher peak currents (ca. 91 %), lower electron transfer resistance (ca. 32-fold) and greater electroactive area (more than twice) when compared to the unmodified ePAD. This significant improvement can be attributed to the increase in the number of active sites available on the electrode surface due to the high conductivity of GNRs. In addition, the ePAD showed good reproducibility and acceptable repeatability, with relative standard deviations (RSDs) ≤ 8.4 % for peak currents and peak potentials. As proof of concept, the GNRs-ePAD was employed for analysis of 5-HTP in pharmaceuticals. As far as we know, this is the first report of the use of ePADs for determination of 5-HTP. The electroanalytical quantification was carried out using square-wave voltammetry (SWV). A linear range from 25 to 1000 μmol L−1 and a limit of detection of 7.6 μmol L−1 were obtained. The sensitivity achieved with the GNRs-ePAD was almost 5-fold greater than that obtained with the bare electrode. The results achieved during the analysis of pharmaceutical products showed good precision (RSD < 3 %) and great concordance with the values found by UV–vis spectrophotometry, utilized for comparison (relative error was < 2 %). None interference or matrix effects were observed during measurements. These results demonstrate promising potential for the GNRs-ePAD proposed here. This device shows itself as an interesting, efficient and low-cost analytical platform for analysis of electroactive species, being specially interesting for places with limited resources.

A Paper-Based Photoelectrochemical Sensing Platform Based on In Situ Grown ZnO/ZnIn2S4 Heterojunctions onto Paper Fibers for Sensitively Detecting AFP.

Nowadays, developing a cost-effective, easy-to-operate, and efficient signal amplification platform is of important to microfluidic paper-based analytical devices (μPAD) for end-use markets of point-of-care (POC) assay applications. Herein, an ultrasensitive, paper-based photoelectrochemical (PEC) bioassay platform is constructed by in situ grown ZnO/ZnIn2S4 heterojunctions onto paper fibers, which acted as photoactive signal amplification probes for enhancing the sensitivity of antibodies-based diagnostic assays, for the sensitive detection of alpha-fetoprotein (AFP) targets. The crystalline flake-like ZnIn2S4 composited with hexagonal nanorods (NRs) morphology of ZnO is an in situ grown, at the first time, onto cellulose fibers surface supported with Au nanoparticle (Au NP) modification to improve conductivity of the device working zone. The obtained composites on paper fibers are implemented as a flexible paper-based photoelectrode to realize remarkable performance of the fabricated μPAD, resulting from the enhanced PEC activity of heterojunctions with effective electron-hole pair separation for accelerating photoelectric conversion efficiency of the sensing process under light irradiation. Once the target AFP was introduced into the biosensing interface assistant, with a specific recognition interaction of AFP antibody, a drastically photocurrent response was generated, in view of the apparent steric effects. With the concentration increase of AFP targets, more immune conjugates could be confined onto the biosensing interface, eventually leading to the quantitative decrease of photocurrent intensity. Combined with an ingenious origami design and permitting the hydrophobic/hydrophilic conversion procedure in the bioassay process, the ultrasensitive PEC detection of AFP targets was realized. Under the optimized conditions, the level of AFP could be sensitively tracked by the prepared μPAD with a liner range from 0.1 to 100 ng mL−1 and limit of detection of 0.03 ng mL−1. This work provides a great potential application for highly selective and sensitive POC testing of AFP, and finally, developments for clinical disease diagnosis.

Recent developments in microfluidic paper-based analytical devices for pharmaceutical analysis.

In the last decade, due to the global increase in diseases, drugs for biomedical applications have increased dramatically. Therefore, there is an urgent need for analytical tools to monitor, treat, investigate, and control drug compounds in diverse matrices. The new and challenging task has been looking for simple, low-cost, rapid, and portable analytical platforms. The development of microfluidic paper-based analytical devices (µPADs) has garnered immense attention in many analytical applications due to the benefit of cellulose structure. It can be functionalized and serves as an ideal channel and scaffold for the transportation and immobilization of various substances. Microfluidic technology has been considered an effective tool in pharmaceutical analysis that facilitates the quantitative measurement of several parameters on cells or other biological systems. The µPADs represent unique advantages over conventional microfluidics, such as the self-pumping capability. They have low material costs, are easy to fabricate, and do not require external power sources. This review gives an overview of the current designs in this decade for µPADs and their respective application in pharmaceutical analysis. These include device designs, choice of paper material, and fabrication techniques with their advantages and drawbacks. In addition, the strategies for improving analytical performance in terms of simplicity, high sensitivity, and selectivity are highlighted, followed by the application of µPADs design for the detection of drug compounds for various purposes. Moreover, we present recent advances involving µPAD technologies in the field of pharmaceutical applications. Finally, we discussed the challenges and potential of µPADs for the transition from laboratory to commercialization.

Microfluidic Paper-Based Device for Medicinal Diagnosis.

The demand for point-of-care testing (POCT) devices has rapidly grown since they offer immediate test results with ease of use, makingthem suitable for home self-testing patients and caretakers. However, the POCT development has faced the challenges of increased cost and limited resources. Therefore, the paper substrate as a low-cost material has been employed to develop a cost-effective POCT device, known as "Microfluidic paper-based analytical devices (μPADs)". This device is gaining attention as a promising tool for medicinal diagnostic applications owing to its unique features of simple fabrication, low cost, enabling manipulation flow (capillarydriven flow), the ability to store reagents, and accommodating multistep assay requirements. This review comprehensively examines the fabrication methods and device designs (2D/3D configuration) and their advantages and disadvantages, focusing on updated μPADs applications for motif identification. The evolution of paper-based devices, starting from the traditional devices of dipstick and lateral flow assay (LFA) with μPADs, has been described. Patterned structure fabrication of each technique has been compared among the equipment used, benefits, and drawbacks. Microfluidic device designs, including 2D and 3D configurations, have been introduced as well as their modifications. Various designs of μPADs have been integrated with many powerful detection methods such as colorimetry, electrochemistry, fluorescence, chemiluminescence, electrochemiluminescence, and SER-based sensors for medicinal diagnosis applications. The μPADs potential to deal with commercialization in terms of the state-of-the-art of μPADs in medicinal diagnosis has been discussed. A great prototype, which is currently in a reallife application breakthrough, has been updated.

A portable tool for colorimetric detection of corrosion inhibitors using paper-based analytical devices

In this work, we report a complete and portable analytical instrumentation dedicated to the colorimetric analysis of corrosion inhibitors. The construction of the analytical tool was planned based on carousel architecture and it involves a simple combination of a 3D-printed holder containing eight circular zones and an Arduino microcontroller photoelectric sensor. In addition, paper-based analytical devices (PAD) were manufactured in the form of microzones using manual cutting and assembled in the 3D printed holder for performing colorimetric measurements of quaternary amines in wastewater samples from the petroleum industry. A blue dye was used as a model color and the obtained pixels responses through the proposed analytical tool and well-known smartphone-based analysis revealed similar colorimetric intensities, thus clearly demonstrating the high performance of the 3D-printed analytical instrumentation. The target colorimetric reaction occurs via the interaction of ammonium quaternary present in corrosion inhibitors composition with the chromogen 3′, 3″, 5′, 5″ – tetrabromophenolphthalein-ethyl-ester (TBPE) in an acidic medium. Detailed optimization of the colorimetric reaction involving the ammonium quaternary was successfully performed, as well as the evaluation of possible physical–chemical interferences (pH, carbonate levels, and temperature) mostly usual in produced water from petroleum pipelines. Recovery studies of ammonium quaternary in real produced water samples exhibited acceptable performance (88 to 121%). Thus, the obtained results successfully demonstrated that the proposed 3D-printed analytical system associated with PADs offers a huge potential to be used for colorimetric detection of corrosion inhibitors in wastewater samples and it can be used in the point-of-need involving the petroleum industry field.

Vanadium-Doped Porous Cobalt Oxide for Its Superior Peroxidase-like Activity in Simultaneous Total Cholesterol and Glucose Determination in Whole Blood Based on a Simple Two-Dimensional Paper-Based Analytical Device.

Vanadium-doped porous Co3O4 (V-porous Co3O4) was synthesized via a simple soft-templating method and used as a superior peroxidase mimic for the simultaneous colorimetric determination of glucose and total cholesterol (TC) in whole blood samples on a two-dimensional microfluidic paper-based analytical device (2D-μPAD). The large surface area and the presence of two metals in V-porous Co3O4 contributed to its excellent catalytic activity toward 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and 3,3',5,5'- tetramethylbenzidine (TMB) with Michaelis-Menten constants (KM) of 0.1301 and 0.0141 mM, respectively. The 2D-μPAD was fabricated using simple wax screen-printing and cutting techniques. The colorimetric reactions of both glucose and TC on 2D-μPAD were simultaneously performed by adding a single drop of a whole blood sample on the sample zone made of the LF1 membrane. After the enzymatic reactions, the generated hydrogen peroxide (H2O2) was oxidized by V-porous Co3O4 to produce hydroxy radicals (•OH), inducing ABTS and TMB to generate colored products. The generated H2O2 was proportional to the intensities of the green and blue products of the glucose and TC systems, respectively. The developed 2D-μPAD required a short analysis time (∼5 min) with small volumes of samples (15 μL of whole blood) whereby no sample preparation was needed. Owing to several advantages including simplicity, low cost, long-term stability, and simultaneous readout, the novel V-porous Co3O4 coupled with 2D-μPAD proved to be promising for practical uses as a pioneering portable device for the determination of glucose, TC, and other important biomarkers without the need of technical supports.

Semi-enclosed paper sensor for highly sensitive and selective detection of proline

In the current study, we have utilized semi-enclosed, leak-proof, microfluidic paper-based analytical devices (μPAD's) modified with isatin conjugated chitosan as specific colorimetric reagent for the detection of proline. Proline is one of the globally accepted stress biomarker in plants and also one of the prominent amino acid present in wine and some processed food. Quantification of proline is regularly required in agriculture field, food and wine industries. Specific interaction of isatin with proline, uniform film forming ability of chitosan which results in uniform coloration and the presence of leak-proof layer which prevent the diffusion of colorimetric reagent deeper resulted in enhancement of color signal intensity at the reaction zone were utilized. Further, the images of the μPAD's were captured using smartphone with 3D printed imaging box which houses smartphone and μPAD's. This platform utilizes smartphone flash for uniform illumination and ensures constant positioning of μPAD's to capture images. This greatly enhances the sensitivity and accuracy of our platform. Compared to previously published highly sensitive multi-layer, paper-based platform for detection of proline, current method has enhanced detection range (∼7 fold) and has comparable limit of detection of 23.75 μM. Moreover, the developed μPAD's platform has reduced optimum reaction temperature and time compared to previous work. The developed paper based platform was utilized for evaluation of proline content in young Arabidopsis plants which are subjected to water stress for 5 days. The devised paper-based methods have the potential to be applicable for the on-site evaluation of various stresses in plants.

RGB-Detector: A Smart, Low-Cost Device for Reading RGB Indexes of Microfluidic Paper-Based Analytical Devices.

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.

Exploitation of N-Gene of SARS-CoV-2 to Develop a New Rapid Assay by ASOs@AuNPs.

A naked-eye (equipment-free), label-free (cost-effective), and RNA extraction-free (to speed up) method for SARS-CoV-2 (as a case study of RNA viruses) detection is developed. Here, the DNA is being used as a template for in situ formation of anisotropic gold nanoparticles (AuNPs) without any chemical modification or DNA labeling. In this study, synthesized AuNPs for the direct detection of N-gene (nucleocapsid phosphoprotein) of SARS-CoV-2 are exploited. To this aim, antisense oligonucleotides (ASOs) with an extra poly guanine tail (G12) were designed. Thus, in the presence of its viral target RNA gene and ASOs@AuNPs-RNA hybridization, there was a red shift in its localized surface plasmon resonance (LSPR), and the intensity of the LSPR peak at 690 nm of throat swab samples was compared to the threshold cycle (Ct) of a reverse-transcriptase real-time polymerase chain reaction (RT-qPCR) (as a gold standard). Results suggested that the plasmonic biosensor can detect a very low amount of SARS-CoV-2 with a detection limit close to RT-qPCR. Simplicity of the new conjugation method with hybridization and annealing without amplification and denaturation steps enabled it to perform in a microfluidic paper-based analytical device.

Colorimetric determination of peroxide value in vegetable oils using a paper based analytical device

Peroxide value (PV) is one of the most typically used quality parameters to monitor lipid oxidation. Here, a simple paper-based analytical device (PAD) has been developed to determine PV in vegetable oils. The analysis is based on setting up the iodometric titration, where hydroperoxides in the oil are reacted with excess iodide ions to generate iodine molecules, on the paper substrate. The device is composed of two paper layers acting as reaction and detection zones, aligned in a metallic mold. A well-defined inverse logarithmic calibration curve was established between the measured PV by the official iodometric method (ISO;3960, 2017) and the color intensity of PAD. It offered a working range of 0.01–30.0 meq/Kg. The limit of detection of 0.015 meq/Kg demonstrated enough sensitivity of the method to estimate peroxide value in edible oils. On-site and visual detection, low cost, simplicity, and less solvent consumption are advantages of the proposed device.

Rapid Assembly of Cellulose Microfibers into Translucent and Flexible Microfluidic Paper-Based Analytical Devices via Wettability Patterning.

Microfluidic paper-based analytical devices (μPADs) are emerging as powerful analytical platforms in clinical diagnostics, food safety, and environmental protection because of their low cost and favorable substrate properties for biosensing. However, the existing top-down fabrication methods of paper-based chips suffer from low resolution (>200 μm). Additionally, papers have limitations in their physical properties (e.g., thickness, transmittance, and mechanical flexibility). Here, we demonstrate a bottom-up approach for the rapid fabrication of heterogeneously controlled paper-based chip arrays. We simply print a wax-patterned microchip with wettability contrasts, enabling automatic and selective assembly of cellulose microfibers to construct predefined paper-based microchip arrays with controllable thickness. This paper-based microchip printing technology is feasible for various substrate materials ranging from inorganic glass to organic polymers, providing a versatile platform for the full range of applications including transparent devices and flexible health monitoring. Our bottom-up printing technology using cellulose microfibers as the starting material provides a lateral resolution down to 42 ± 3 μm and achieves the narrowest channel barrier down to 33 ± 2 μm. As a proof-of-concept demonstration, a flexible paper-based glucose monitor is built for human health care, requiring only 0.3 μL of sample for testing.

Quantum dots immobilized paper for specific and sensitive quantitation of Ni(II) by headspace photochemical vapor generation: Mechanism and application for RGB detection in tea fusion with a smartphone

To avoid the interferences of coexisting ions from sample matrixes during CdTe quantum dots (QDs) based fluorescence and colorimetric assay, an instrument-free paper-based analytical device immobilized CdTe QDs was developed for specific and sensitive quantitation of Ni(II) by headspace photochemical vapor generation (HS-PVG). In the PVG procedure, Ni(II) was converted to volatile Ni(CO)4 by reacting with formic acid under ultraviolet light irradiation, which can effectively quench the fluorescence of the CdTe QDs coated paper. The quenching mechanism was studied in depth via both experimental and theoretical investigations. The results of X-Ray diffraction, X-ray photoelectron spectroscopy, and density functional theory (DFT) show that the generated Ni(CO)4 was reacted with O2 to decompose into NiO at room temperature, which was deposited on the surface of CdTe QDs coated paper, causing its fluorescence quenching. Moreover, visual detection of Ni(II) was achieved with a smartphone by identifying the RGB value of QDs coated paper via HS-PVG procedure. A limit of detection (LOD) of 6.2 μg L−1 was required, which can match those obtained by other large-scale instruments. The concentration of Ni(II) in a series of tea infusions was accurately detected with different leaching experiments and soaking conditions.

Multi-parameter photoelectric data fitting for microfluidic sweat colorimetric analysis

Colorimetric detection has been widely used in sweat colorimetric analysis due to its advantages of convenient detection and easy implementation. However, its relatively low detection accuracy limits its application. It is therefore necessary to develop new methods to extract quantitative data from the colorimetric biosensor. In this work, a plug type paper-based analytical device (μPad) sensor was fabricated and put into a Polydimethylsiloxane (PDMS) microfluidic chamber prepared through 3D-Printing. Photoelectric sensor with a light-emitting diode (LED) light source was utilized for acquiring the colour change information (including R, G, B, Lux, colour temperature value) before and after the colorimetric sensing. Wireless Bluetooth transmission module was used for data acquisition. The obtained multi-parameter data was fitted through machine learning algorithm using Python to establish the relationship between the multi-parameter and the concentration of analyte. Discoloration experiments of glucose, lactic acid and pH were used to verify the feasibility of the sensor and data analysis method. Analysis of human sweat in a running volunteer has proved its potential as a wearable device and provided a new insight for data analysis of colorimetric method of wearable colorimetric biosensors.

Microfluidic paper-based analytical device for determination of sucrose in sugarcane juice using Benedict's reagent.

This work presents a microfluidic paper-based analytical device (μPAD) for the determination of sucrose using the Benedict's test. An asymmetric dumbbell-shaped hydrophobic barrier was produced by rubber stamping the barrier pattern onto a laboratory filter paper. Hydrochloric acid and solution containing sucrose were successively deposited onto the sample reservoir of the μPAD attached to a glass slide. The device was placed in a plastic bag and dipped into boiling water for accelerating the hydrolysis of sucrose into the reducing sugars. Then the Benedict's reagent was added at the narrow straight channel connecting the two circular zones of the μPAD, which was replaced in the plastic bag and heated again for reduction of Cu(II) by the reducing sugars. Precipitate of brick-red copper(I) oxide was formed. The image of the μPAD was recorded by a smartphone. The ratio of the red to blue intensities gave linear correlation with the concentration of sucrose in the range of 0.5-10% w/v. The relative standard deviation of the measurement was less than 5% for 2 and 4% w/v sucrose (n = 10), with limit of determination, calculated using standard deviation of regression divided by slope of calibration, of 0.26% w/v sucrose. The method was successfully validated using the dinitrosalicylic acid method for sucrose measurement. Percent recoveries of sucrose were evaluated using ten sugarcane samples. The recoveries were in the range of 89 to 101%, demonstrating that there were no significant sample matrix effects on the quantification.

Integrated calibration and serum iron in situ analysis into an array microfluidic paper-based analytical device with smartphone readout

In this work, a colorimetric microfluidic paper-based analytical device (μPAD) combined with a smartphone readout was proposed for the determination of serum iron (Fe3+), which is linked to transferrin. Firstly, Fe3+ was selectively isolated and preconcentrated from serum by using anti-transferrin immunomagnetic beads (anti-Tf-MBs). Secondly, Fe3+ is reduced to Fe2+ by a hydroxylamine solution (pH 4.8) and then measured in the μPAD, which contains the colorimetric reagent ferrozine. Finally, the intensity of the purple color formed in the μPAD was measured by a smartphone. The approach exhibited an excellent linear correlation (r = 0.996) and good limit of detection (0.3 μg mL−1). Moreover, a certified reference material (human serum) was analyzed by this approach, showing an excellent accuracy (Er < 4%) and inter-device reproducibility (RSD = 1%, n = 3).Interestingly, the μPAD array-design allowed the simultaneous analysis of different samples, improving the sample throughput (up to 5 samples in 130 min, using 100 μL each), and the integration of calibration and analysis into the same device, simplifying the analysis without losing accuracy or sensitivity, and avoiding inter-device variability, which constituted an added value to this approach.These disposable μPADs meet several requirements of point-of-care testing (POCT) because it is cheap, portable, easy-to-use, sensitive, and specific. Therefore, it may be an interesting way for measuring patients’ serum iron levels in situ with reliability, especially, in developing countries, where the prevalence of iron deficiency and iron-deficiency anemia is higher and there are lower health resources than in developed countries.

Electric yo-yo centrifugation combining with paper-based microfluidic immunoassay chip for inflammatory biomarkers detection in whole blood

At present, most countries or regions use commercial centrifuges for centrifugation, but this is out of reaching for limited-resource areas. To overcome this problem, a portable electric yo-yo as centrifuge was firstly proposed to obtain serum, and this device can be combined with paper-based analytical devices for enzyme-linked immunosorbent assay (ELISA) analysis from human whole blood. In this study, inflammatory biomarkers C-reactive protein (CRP) and serum amyloid A (SAA) were used as target biomarker to verify the performance of the proposed method. The results shows good performance and their detection limits were determined to be 580 pg/mL for CRP and 800 pg/mL for SAA, respectively. We believe this method provides a new platform of low cost and fast detection for inflammatory biomarkers in the limited-resource settings.

Colorimetric detection of H2O2 with Fe3O4@Chi nanozyme modified µPADs using artificial intelligence.

Peroxidase mimicking Fe3O4@Chitosan (Fe3O4@Chi) nanozyme was synthesized and used for high-sensitive enzyme-free colorimetric detection of H2O2. The nanozyme was characterized in comparison with Fe3O4 nanoparticles (NPs) using X-ray diffraction,Fourier-transform infrared spectroscopy,dynamic light scattering, and thermogravimetric analysis. The catalytic performance of Fe3O4@Chi nanozyme was first evaluated by UV-Vis spectroscopy using 3,3',5,5'-tetramethylbenzidine. Unlike Fe3O4NPs, Fe3O4@Chi nanozyme exhibited an intrinsic peroxidase activity with a detectionlimit of 69nM. Next, the nanozyme was applied to a microfluidic paper-based analytical device (µPAD) and colorimetric analysis was performed at varying concentrations of H2O2 using a machine learning-based smartphone app called "Hi-perox Sens++ ." The app with machine learning classifiers made the system user-friendly as well as more robust and adaptive against variation in illumination and camera optics. In order to train various machine learning classifiers, the images of the µPADs were taken at 30s and 10min by four smartphone brands under seven different illuminations. According to the results, linear discriminant analysis exhibited the highest classification accuracy (98.7%) with phone-independent repeatability at t = 30s and the accuracy was preserved for 10min. The proposed system also showed excellent selectivity in the presence of various interfering molecules and good detection performance in tap water.

Palladium-platinum bimetallic nanomaterials and their application in Staphylococcus aureus detection on paper-based devices

In this study, we demonstrate how palladium@platinum nanoparticles (Pd@Pt NPs) can be used as a nanozyme for the detection of Staphylococcus aureus (S. aureus) on a paper-based analytical device. Pt was doped with Pd to form bimetallic NPs that serve as a peroxidase mimetic, which can enhance the reaction area with a substrate (3, 3', 5, 5'-tetramethylbenzidine, TMB). After material characterization, we show the peroxidase-like activity of the Pd@Pt NPs featured a 13-times higher binding affinity value (Km) for TMB compared to horseradish peroxidase, which is currently widely used in immunoassays. By incorporating the Pd@Pt NPs in a paper-based immunoassay, we can detect protein A, the biomarker of S. aureus, within 30 min with a detection limit of 9.56 ng/mL. We have also successfully validated this nanozyme-immobilized paper-based analytical device (nPAD) for the detection of human immunoglobulin G to demonstrate the capability of the Pd@Pt NPs for different target analytes. This highly sensitive, rapid, and portable nPAD design has the potential for personalized medicine and point-of-care testing, which could expand on-site prognoses in resource-limited settings.

Paper microfluidic device and spectrophotometry methods for colorimetric detection of phenylephrine hydrochloride in pure and pharmaceutical preparations

ABSTRACT. Two easy-to-use, rapid, and inexpensive colorimetric-based assays for the detection of active constituent in nasal decongestant phenylephrine hydrochloride (PHP), in solution and their respective pharmaceutical preparations were reported. The assay principle depends on the diazotized sulfadimidine (DSDM) coupling with PHP active pharmaceutical compound under alkaline conditions to produce a colored azo-dye product. A UV/VIS spectrophotometry method was utilized to monitor the color absorbance change at 460 nm, with a linear calibration graph in the concentration range of 1-20 mg L-1 and a limit of detection of 0.22 mg L-1 and limit of quantification of 0.72 mg L-1. A comparison was done using a paper-based microfluidic analytical device (µPADs) and color intensity change was measured using a Samsung note 9 camera to capture digital images and Image J software to analyze the color intensity development. A linear trend was observed above approximately 10 mg L-1. Thus, paper-based microfluidic analytical devices (µPADs) can be used as an alternative platform to conventional methods to perform a semi-quantitative PHP drug analysis. The proposed method was applied for the determination of PHP in commercial pharmaceutical preparations.&#x0D; &#x0D; KEY WORDS: Phenylephrine hydrochloride, Spectrophotometry, Paper-based microfluidic analytical devices (µPADs), Pharmaceutical preparations, Colorimetric detection&#x0D; &#x0D; Bull. Chem. Soc. Ethiop. 2022, 36(4), 727-736. &#x0D; DOI: https://dx.doi.org/10.4314/bcse.v36i4.1