Simple distance-based thread analytical device integrated with ion imprinted polymer for Zn2+ quantification in human urine samples.

This article presents the development of a distance-based thread analytical device (dTAD) integrated with an ion-imprinted polymer (IIP) for quantitative monitoring of zinc ions (Zn2+) in human urine samples. The IIP was easily chemically modified onto the thread channel using dithizone (DTZ) as a ligand to bind to Zn2+ with methacrylic acid (MAA) as a functional monomer and ethylene glycol dimethacrylate (EGDMA) as well as 2,2-azobisisobutyronitrile (AIBN) as cross-linking agents to enhance the selectivity for Zn2+ detection. The imprinted polymer was characterized using Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy and Scanning Electron Microscopy-Energy Dispersive X-ray Spectroscopy (SEM-EDS). Under optimization, the linear detection range was from 1.0 to 20.0 mg L-1 (R2 = 0.9992) with a limit of detection (LOD) of 1.0 mg L-1. Other potentially interfering metal ions and molecules did not interfere with this approach, leading to high selectivity. Furthermore, our technique exhibits a remarkable recovery ranging from 100.48% to 103.16%, with the highest relative standard deviation (% RSD) of 5.44% for monitoring Zn2+ in human control urine samples, indicating high accuracy and precision. Similarly, there is no significant statistical difference between the results obtained using our method and standards on zinc supplement sample labels. The proposed method offers several advantages in detecting trace Zn2+ for point-of-care (POC) medical diagnostics and environmental sample analysis, such as ease of use, instrument-free readout, and cost efficiency. Overall, our developed dTAD-based IIP method holds potential for simple, affordable, and rapid detection of Zn2+ levels and can be applied to other metal ions' analysis.

Distance-based paper analytical devices integrated with molecular imprinted polymers for Escherichia coli quantification.

The development of distance-based paper analytical devices (dPADs) integrated with molecularly imprinted polymers (MIPs) to monitor Escherichia coli (E. coli) levels in food samples is presented. The fluidic workflow on the device is controlled using a designed hydrophilic bridge valve. Dopamine serves as a monomer for the formation of the E. coli-selective MIP layer on the dPADs. The detection principle relies on the inhibition of the E. coli toward copper (II) (Cu2+)-triggered oxidation of o-phenylenediamine (OPD) on the paper substrate. Quantitative detection is simply determined through visual observation of the residual yellow color of the OPD in the detection zone, which is proportional to E. coli concentration. The sensing exhibits a linear range from 25.0 to 1200.0 CFU mL-1 (R2 = 0.9992) and a detection limit (LOD) of 25.0 CFU mL-1 for E. coli detection. Additionally, the technique is highly selective with no interference even from the molecules that have shown to react with OPD to form oxidized OPD. The developed device demonstrates accuracy and precision for E. coli quantification in food samples with recovery percentages between 98.3 and 104.7% and the highest relative standard deviation (RSD) of 4.55%. T-test validation shows no significant difference in E. coli concentration measured between our method and a commercial assay. The proposed dPAD sensor has the potential for selective and affordable E. coli determination  in food samples without requiring sample preparation. Furthermore, this strategy can be extended to monitor other molecules for which MIP can be developed and integrated into paper-microfluidic platform.

Distance-based paper analytical device for multiplexed quantification of cytokine biomarkers using carbon dots integrated with molecularly imprinted polymer.

This article introduces distance-based paper analytical devices (dPADs) integrated with molecularly imprinted polymers (MIPs) and carbon dots (CDs) for simultaneous quantification of cytokine biomarkers, namely C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) in human biological samples for diagnosis of cytokine syndrome. Using fluorescent CDs and MIP technology, the dPAD exhibits high selectivity and sensitivity. Detection is based on fluorescence quenching of CDs achieved through the interaction of the target analytes with the MIP layer on the paper substrate. Quantitative analysis is easily accomplished by measuring the distance length of quenched fluorescence with a traditional ruler and naked eye readout enabling rapid diagnosis of cytokine syndrome and the underlying infection. Our sensor demonstrated linear ranges of 2.50-24.0 pg mL-1 (R2 = 0.9974), 0.25-3.20 pg mL-1 (R2 = 0.9985), and 1.50-16.0 pg mL-1 (R2 = 0.9966) with detection limits (LODs) of 2.50, 0.25, and 1.50 pg mL-1 for CRP, TNF-α, and IL-6, respectively. This sensor also demonstrated remarkable selectivity compared to a sensor employing a non-imprinted polymer (NIP), and precision with the highest relative standard deviation (RSD) of 5.14%. The sensor is more accessible compared to prior methods relying on expensive reagents and instruments and complex fabrication methods. Furthermore, the assay provided notable accuracy for monitoring these biomarkers in various human samples with recovery percentages ranging between 99.22% and 103.58%. By integrating microfluidic systems, nanosensing, and MIPs technology, our developed dPADs hold significant potential as a cost-effective and user-friendly analytical method for point-of-care diagnostics (POC) of cytokine-related disorders. This concept can be further extended to developing diagnostic devices for other biomarkers.

Photothermal biosensing integrated with microfluidic paper-based analytical device for sensitive quantification of sarcosine.

This article presents the development of a photothermal biosensing integrated with microfluidic paper-based analytical device (PT-µPAD) as a quantitative biosensor method for monitoring sarcosine in human control urine, plasma, and serum samples. The device utilizes gold nanoparticles (AuNPs) as both a peroxidase-like nanozyme and a photothermal substrate to enable sarcosine detection. In our PT-µPAD, hydrogen peroxide (H2O2) is generated through the oxidation of sarcosine by a sarcosine oxidase (SOx) enzyme. Subsequently, the H2O2 flows through the paper microchannels to the detection zone, where it etches the pre-deposited AuNPs, inducing a temperature change upon exposure by a 532 nm laser. The temperature variation is then measured using a portable and inexpensive infrared thermometer. Under optimized conditions, we obtained a linear range between 10.0 and 40.0 nmol L-1 (R2 = 0.9954) and a detection limit (LOD) of 32.0 pmol L-1. These values fall within the clinical range for sarcosine monitoring in prostate cancer diagnostics in humans. Moreover, our approach exhibits high selectivity without interfering effects. Recovery studies in various human control samples demonstrated a range of 99.05-102.11 % with the highest RSD of 2.25 %. The PT-µPAD was further validated for sarcosine determination in human control urine and compared with a commercial ELISA assay, revealing no significant difference between these two methods at a 95 % confidence level. Overall, our proposed sarcosine biosensor is well-suited for prostate cancer monitoring, given its affordability, sensitivity, and user-friendliness, even for unskilled individuals. Moreover, this strategy has promising prospects for broader applications, potentially detecting various biomarkers as a point-of-care (POC) diagnostic tool.

Gold Nanomaterial-Based Microfluidic Paper Analytical Device for Simultaneous Quantification of Gram-Negative Bacteria and Nitrite Ions in Water Samples.

This study presents a rapid microfluidic paper-based analytical device (µPAD) capable of simultaneously monitoring Gram-negative bacteria and nitrite ions (NO2-) for water quality monitoring. We utilize gold nanoparticles (AuNPs) functionalized with polymyxin molecules (AuNPs@polymyxin) to cause color change due to aggregation for the detection of Gram-negative bacteria, and antiaggregation in the presence of o-phenylenediamine (OPD) for NO2- detection. In this study, Escherichia coli (E. coli) serves as the model of a Gram-negative bacterium. Using the developed µPADs, the color changes resulting from aggregation and antiaggregation reactions are measured using a smartphone application. The linear detection ranges from 5.0 × 102 to 5.0 × 105 CFU/mL (R2 = 0.9961) for E. coli and 0.20 to 2.0 µmol/L (R2 = 0.995) for NO2-. The detection limits were determined as 2.0 × 102 CFU/mL for E. coli and 0.18 µmol/L for NO2-. Notably, the newly developed assay exhibited high selectivity with no interference from Gram-positive bacteria. Additionally, we obtained acceptable recovery for monitoring E. coli and NO2- in drinking water samples with no significant difference between our method and a commercial assay by t test validation. The sensor was also employed for assessing the quality of the pond and environmental water source. Notably, this approach can also be applied to human urine samples with satisfactory accuracy. Furthermore, the assay's stability is extended due to its reliance on AuNPs rather than reagents like antibodies and enzymes, reducing costs and ensuring long-term viability. Our cost-effective µPADs therefore provide a real-time analysis of both contaminants, making them suitable for assessing water quality in resource-limited settings.

A sensitive and facile electrochemical paper-based sensor for glucose detection in whole blood using the Pd/CB-Ni@rGO modified electrode.

We created novel Pd/CB-Ni@rGO nanomaterials for glucose detection. The as-synthesized nanomaterials were dropped on the electrode surface using the drop casting technique. The prepared electrode was then attached to a paper-based device containing the sample zone and the reaction zone, enabling plasma isolation and an enzymatic reaction for glucose detection in whole blood. The nanomaterials and surfaces of electrodes were characterized by FTIR, TEM, and SEM. The proposed approach is a disposable glucose detection method that is unaffected by protein fouling on the electrode, and it requires only one drop of human blood. Therefore, there is no need for extensive sample preparation, and there is less sample consumption. Under optimal conditions, Pd/CB-Ni@rGO can accurately measure blood glucose levels with a linear range of 7 to 7140 µM (R2 = 0.9986) and a low detection limit of 0.82 µM. Besides, the developed sensor shows excellent anti-interference capacity, stability, and satisfactory reproducibility and repeatability. Importantly, Pd/CB-Ni@rGO was successfully applied for glucose in whole blood from 4 volunteers, with results that correlated well with those obtained using an Accucheck glucometer at a 95% confidence level. Given its low cost, high accuracy, and ease of use, the blood glucose sensor holds significant potential for clinical use and broadens the area of future noninvasive sensor development.

Highly Sensitive Photothermal Microfluidic Thread-Based Duplex Immunosensor for Point-of-Care Monitoring.

Herein, we successfully developed a thread-based analytical device (µTAD) for simultaneous immunosensing of two biomolecules with attomolar sensitivity by using a photothermal effect. A photothermal effect exploits a strong light-to-heat energy conversion of plasmonic metallic nanoparticles at localized surface plasmon resonance. The key innovation is to utilize the cotton thread to realize this sensor and the use of chitosan modification for enhancing the microfluidic properties, for improving the efficiency of photothermal conversion, and for sensor stability. The developed µTAD sensor consists of (i) a sample zone, (ii) a conjugation zone coated with gold nanoparticles bound with an antibody (AuNPs-Ab2), and (iii) a test zone immobilized with a capture antibody (anti-Ab1). The prepared µTAD is assembled in a custom three-dimensional (3D) printed device which holds the laser for illumination and the thermometer for readout. The 3D-printed supportive device enhances signal response by focusing light and localizing the heat generated. For proof of concept, simultaneous sensing of two key stress and inflammation biomarkers, namely, cortisol and interleukin-6 (IL-6), are monitored using this technique. Under optimization, this device exhibited a detection linear range of 2.0-14.0 ag/mL (R2 = 0.9988) and 30.0-360.0 fg/mL (R2 = 0.9942) with a detection limit (LOD) of 1.40 ag/mL (∼3.86 amol/L) and 20.0 fg/mL (∼950.0 amol/L) for cortisol and IL-6, respectively. Furthermore, the analysis of both biomolecules in human samples indicated recoveries in the range of 98.8%-102.88% with the highest relative standard deviation being 3.49%, offering great accuracy and precision. These results are the highest reported sensitivity for these analytes using an immunoassay method. Our PT-µTAD strategy is therefore a promising approach for detecting biomolecules in resource-limited point-of-care settings.

Nanomaterials integrated with microfluidic paper-based analytical devices for enzyme-free glucose quantification.

In this study, nanomaterials capable of enzyme-free glucose quantification and colorimetric readout are integrated into a microfluidic paper-based analytical devices (µPADs). Gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs) were utilized as a peroxidase-like nanozyme and a colorimetric probe to achieve glucose monitoring. In this developed device, glucose is oxidized by AuNPs to generate hydrogen peroxide (H2O2), which flows in the paper microchannels toward detection zones. H2O2 then etches the immobilized AgNPs to induce a color change. The intensity of color change is easily monitored using a smartphone application. Following method optimization, we obtained a linear range from 0.50 to 10.0 mmol L-1 (R2 = 0.9921) and a detection limit (LOD) of 340.0 µmol L-1. This falls in the clinically relevant range for glucose monitoring and diabetes diagnosis in humans. In addition, the total analysis time is just 20 min, which is significantly less than the same experiment performed in the solution phase. Also, our method is markedly selective; other substrates do not interfere. The recovery test in human control samples was in the range of 98.47-102.34% and the highest relative standard deviation (RSD) was 3.58%. The enzyme-free approach for glucose sensing is highly desirable for diabetes diagnosis as it replaces the more expensive enzyme with cheaper nanomaterials. Furthermore, since nanomaterials are more environmentally stable compared to enzymes, it has the potential for widespread deployment as point-of-care diagnostics (POC) in resource-limited settings.

Distance-Based All-In-One Immunodevice for Point-of-Care Monitoring of Cytokine Interleukin-6.

We report the development of a distance-based paper analytical device combined with a hydrophilic bridge valve (B-dPAD) as a quantitative immunoassay method to monitor human interleukin-6 (IL-6) in human samples. Our device design features (i) a circular sample inlet zone, (ii) a circular capture zone with immobilized anti-IL-6 (anti-Ab1), and (iii) a detection zone channel coated with methylene blue (MB). Two hydrophilic valves are positioned between these three zones. IL-6 levels were determined quantitatively by measuring the extent of degradation of MB to a colorless product along the length of the detection zone channel. Following method optimization, we obtained a linear range from 0.05 to 25.0 pg/mL (R2 = 0.9995) and a detection limit (LOD) of 0.05 pg/mL by the naked-eye readout. This is directly within the clinically relevant range. The system does not require any external instrumentation, and the bridge valves can be easily connected and disconnected by a minimally trained operator. The total analysis time is 35 min, significantly reduced from a typical ELISA assay, which takes around 1 h since the B-dPAD workflow circumvents washing steps. The device was tested for IL-6 quantification in human saliva and urine samples of volunteers, with no significant difference found between our method and the standard clinical laboratory method at 95% confidence levels. Recoveries ranged from 98 to 105% with the highest standard deviation at 3.9%. Our B-dPAD immunodevice is therefore a promising approach for rapid IL-6 monitoring in the context of point-of-care diagnostics and analysis in resource-limited settings.

A Simple Counting-Based Measurement for Paper Analytical Devices and Their Application.

This work introduces the concept of a counting-based measurement on paper analytical devices (cPADs) to improve the utilization of numerous reactions. The design of cPADs consists of two layers of paper substrates; the first layer contains a central sample zone combined with a radial surrounded by 12 detection zones that are predeposited with the various reagents, and the second layer acts as a connection channel between the sample zone and each detection zone. The solution can vertically flow from the first to the second layer and then move through the area to each subsequent detection zone. The analyte level can be evaluated by counting the number of detection zones that change color from a blank signal. Furthermore, our cPADs exhibit a capability of implementation for a broad series of reactions. Compared to the dPAD technique, some reactions that are possibly difficult to apply in such devices can be wholly enabled in our devices. The final color reaction on cPADs can apparently occur due to its identity. We applied this technique to the monitoring of carbaryl (CBR) and copper ions (Cu2+) using different reactions, including azo-coupling and complexation, respectively. Accordingly, this indicates an excellent result validated using the more traditional methods. Our cPADs can be applied for rapid screening of both CBR and Cu2+ in water samples with outstanding accuracy and precision using a naked-eye measurement by a relatively unskilled person. We offer a simple platform on PADs for rapid screening, combining high cost-effectiveness within a miniaturized platform designed for use with onsite applications, which is thus suitable for several different reactions.

Distance-based ß-amyloid protein detection on PADs for the scanning and subsequent follow-up of Alzheimer's disease in human urine samples.

We report on the first development of a simple distance-based ß-amyloid (Aß) protein quantification using a paper-based device (dPAD) to screen for Alzheimer's disease (AD) and to subsequently follow up on its influence, i.e., clinical dementia. This sensor method is based on the transformation of a free acid form and its binding with a basic form of bromocresol purple (BCP) through its electrostatic interaction with an Aß protein. This sensor can measure the length of color change from yellow to blue-green on a paper strip, with this change proportional to the amount of Aß protein level. We found that the linearity for Aß protein monitoring was in the range from 0.50 to 10.0 ng mL-1, and the subsequent naked-eye detection limit for Aß was 0.20 ng mL-1. This system also provided high reproducibility and with no apparent interference effect for Aß protein analysis in human urine samples. Furthermore, our developed dPAD constituted an accurate and effective device to precisely determine an Aß protein concentration in real samples, with percentage recoveries in the range of 97-103%, and with the highest relative standard deviation of 5.41%. Subsequently, the validation of our assay was assessed by comparison with a commercial ELISA approach, with favorable results. Finally, the proposed dPAD was successfully applied to the determination of an Aß protein in human urine samples and showed more benefits for the unskilled user, such as cost-efficiency, simplicity, low reagent usage, and low time consumption. It is also suitable for point-of-care monitoring.

Distance-Based Paper Device for a Naked-Eye Albumin-to-Alkaline Phosphatase Ratio Assay.

The albumin-to-alkaline phosphatase ratio (AAPR) has been a cancer prognostic indicator. This paper presents the concept of a dual-color change distance-based paper device (dPAD) for albumin (Alb) and alkaline phosphatase (ALP) detection to evaluate this cancer prognostic index. Whereas Alb interacts with the bromocresol green (BCG) indicator to form a bluish-green complex, ALP hydrolyzes l-ascorbic acid-2-phosphate (AAP) to produce ascorbic acid (AA), which reacts with KIO3 to generate I2 and I-. I2/I- reacts with silver hexagonal nanoprisms (purple color) in the presence of Cu2+, resulting in a color change from purple to colorless. The distance of the color change from yellow to the bluish-green and purple to colorless correlates to Alb and ALP concentration, respectively. The angle index for the AAPR is then defined by drawing a straight line that connects the tops of the two changed band lengths in the detection area. The highest bluish-green color band length on the Alb region is the midpoint, which is the position set of the protractor at 0°, and the angle is measured using a simple protractor. The results indicate that an AAPR below 0.57 will have an angle greater than 40° and correlates with a risk factor for lung cancer. The naked-eye detection limits for Alb and ALP were found to be 0.8 g/L and 5 U/L (n = 10), respectively. The practical application of the developed dPAD was successfully demonstrated by Alb and ALP analysis in human serum and validated against standard methods. The proposed method does not require incubation conditions for the ALP assay, which strongly reduces the overall analysis steps and time. Moreover, our device provides a low-cost, simple, sensitive, selective, accurate, and precise determination of the AAPR.

A rapid and highly sensitive paper-based colorimetric device for the on-site screening of ammonia gas.

A rapid and highly sensitive paper-based colorimetric device for the on-site detection of ammonia (NH3) gas is presented in this study. The detection principle of this device is based upon a change of color from red to yellow on a paper that has been immobilized with a pH indicator, i.e., methyl orange (pKa = 3.4), in the presence of NH3 gas. The color signal of the device can be measured through the hue channel of an HSL system via the application of a smartphone. This device can detect the amount of NH3 gas within 3 min. The linear relationship between the NH3 gas concentration and the hue signal was found to be in the range from 6.0 to 54.0 ppbv with R2 = 0.9971, and the limit of detection was found to be 2.0 ppbv. In addition, this device showed remarkably high selectivity to NH3 gas amongst the other common volatile organic compounds and general gases that are present in environmental air without the assistance of any membrane material. Furthermore, we demonstrated the applicability of this device for the detection of total NH3 gas at a chicken farm and in a laboratory, with relative standard deviations of 6.2% and 5.4%, respectively. The developed NH3 gas device in the study is easy to operate and cost-effective, with the reduction of a large consumption of chemical reagents; also, its signals can be measured simply and then recorded through a smartphone. It is suitable for the application of routine on-site detection of NH3 gas, especially concerning regions which have limited resources.

Highly sensitive distance-based liquid crystalline visualization for paper-based analytical devices.

We successfully report on the first demonstration of a highly sensitive distance-based liquid crystalline visualization for paper-based analytical devices. The construction of this paper sensor was achieved by immobilizing 4-cyano-4'-pentylbiphenyl (5CB) as liquid crystalline molecules (LCs) onto a paper strip substrate. The sensing mechanism is based on the ultrasound-assisted decomposition of 5CB by the hydroxyl radical (•OH) which is generated from the oxidase enzymatic reaction of the analyte, this then results in the change of texture and color of paper. The utility of our devices was then demonstrated with the determination of bilirubin (BR) in biological samples using a bilirubin oxidase enzymatic reaction. The quantification of BR can be achieved by dipping the tip of the paper strips into the analyte solutions and then by measuring the length of color which has been changed on the paper, by the naked eye. Under optimized conditions, this paper sensor offered the linear range of BR detection from 2.0 to 30.0 pmol/L (R2 = 0.9945) with the limit of detection (LOD) of 0.80 pmol/L. In addition, the results of this sensor were highly reproducible, with a relative standard deviation (RSD) of less than 3.50%. The recoveries of spiked BR in human urine and serum samples were in the range of 99.09-107.89%, which demonstrates the high accuracy of this paper sensor. Overall, this work presents a simple method to determine the concentration of H2O2 and BR at pmol levels with an instrument-free length-measuring readout, so it could be suitable for quantitative analysis of other biomarkers based on oxidase enzymatic reaction, which can provide important information about early disease diagnosis and patient prognosis.

Rapid Distance-Based Cardiac Troponin Quantification Using Paper Analytical Devices for the Screening and the Follow-Up of Acute Myocardial Infarction, Using a Single Drop of Human Whole Blood.

This work introduces the procedure of using non-immunoassay distance-based paper analytical devices (dPADs) to accurately measure any traces of the cardiac troponin I (TnI) in whole blood samples without the use of any external blood separation. This enables a rapid clinical diagnosis and the subsequent follow-up in regard to identifying acute myocardial infarction. These dPADs are designed and constructed to accommodate three parts: (1) a blood separation zone that is immobilized with a hemostatic agent, this no longer requires a blood separation membrane for the isolation of the plasma from the blood element, (2) a pretreatment zone, and (3) a detection zone coated with thymol blue. The quantitative TnI level in the whole blood was determined by measuring the blue color length found in the detection zone, which is proportional to the concentration, owing to the dry protein binding principle. Correspondingly, a mere single drop of human whole blood performs adequately within our proposed method. This reduces both the size of the collection process and the sample volumes needed in the respective medical fields. As we cover all of the optimization studies, our dPADs provide an evaluation of the linearity range from 0.025 to 2.5 ng/mL (R2 = 0.9989) of TnI, with a detection limit as low as 0.025 ng/mL by use of an observation just using the naked eye. To validate the clinical utilities of our proposed method, our dPADs were then applied for the detection of TnI in humans using the whole blood sample of 15 volunteers. A great amount of accuracy was required in this assay because there was no significant difference between both methods, with the confidence level being as high as 95%. This technique also showed that the recoveries ranged from 99.40 to 104.27%, with the highest relative standard deviation being at 3.77%. Thus, our proposed dPADs offer more benefits for a rapid TnI determination.

A Portable Reflective Absorbance Spectrophotometric Smartphone Device for the Rapid and Highly Accurate Determination of Amlodipine in Pharmaceutical Formulation and Human Urine Samples.

The simple reflective absorbance spectrophotometric smartphone device for point-of-monitoring amlodipine is presented here for the first time. The immediate analysis of amlodipine in the human urine of the patients who suffered severe side effects of this drug is very important for the diagnosis, treatment, and reduction of the death rate. This measurement technique is based on the charge-transfer complex between amlodipine and picric acid, which forms a yellow product. This product can absorb light intensity from an LED strip and measure through the Blue channel from the RGB mode with a smartphone application. The linear relationship for amlodipine monitoring was found in a wide range from 100.0 µg L-1 to 140.0 mg L-1 (R2 = 0.999), and the limit of detection was found to be 25.0 µg L-1. Our proposed method can be applied to different smartphone brands with consistent sensitivity of amlodipine detection. Additionally, the determination of amlodipine in pharmaceutical formulations and human urine samples was demonstrated by our proposed method. The recoveries were indicated in the range of 98.60 - 100.00%, which is at the acceptable level for pharmacy. This method offers an interweaving of basic technology and chemical analysis with being environmentally friendly due to reducing the complex instrument and the amount of organic waste compared to the chromatographic technique and efficient use for the detection of amlodipine. Hence, this method can be applied for prompt medical diagnoses and laboratories with limited budget resources.

Development of an ultrasound-enhanced smartphone colorimetric biosensor for ultrasensitive hydrogen peroxide detection and its applications.

In this work, we developed the first ultrasound technique enhanced smartphone application for highly sensitive determination of hydrogen peroxide (H2O2). The measurement technique is based on the change in color intensity due to the transformation of tetramethylbenzidine (TMB) to oxidized tetramethylbenzidine (oxTMB) by the oxidation process with hydroxyl radical (OH˙) from the oxidation etching of silver nanoparticles (AgNPs) and its ultrasound usability. The oxTMB product occurs without peroxidase and can be detected with a saturation channel using HSV methodology via the application of a smartphone. To prove the peroxidase mimic property, our proposed method was also validated by determination of certain biomolecules, including glucose, uric acid, acetylcholine and total cholesterol, of which the known amounts are a valuable diagnostic tool. The proposed method provided the lowest limits of detection (LOD) of 2.0, 5.0, 12.50, 7.50, and 10.0 nmol L-1 for H2O2, glucose, uric acid, acetylcholine, and cholesterol, respectively, when compared with LODs obtained from other smartphone colorimetric methods. Reproducibility was calculated from the detection of H2O2 at 25.0 and 50.0 nmol L-1 with the highest standard deviations of 3.47 and 4.58%, respectively. Additionally, the determination of all analytes in human urine samples indicated recoveries in the range of 96-104% with the highest relative standard deviation of 3.98%, offering high accuracy and precision. Our research shows the novel compatibility of basic technology and chemical methodology with green chemistry principles by reducing a high-power process and organic solvent as well as exhibiting good colorimetric performance and effective sensitivity and selectivity. Thus, our developed method can be applied for point-of-care medical diagnosis.