POC device for rapid oral pH determination based on a smartphone platform.

Salivary pH serves as a valuable and useful diagnostic marker for periodontal disease, as it not only plays a critical role in disease prevention but also in its development. Typically, saliva sampling is collected by draining and spitting it into collection tubes or using swabs. In this study, we have developed a Point-of-Care (POC) device for in situ determination of oral pH without the need for complex instruments, relying solely on a smartphone as the detection device. Our system utilizes a non-toxic vegetable colourimetric indicator, immobilized on a chitosan membrane located on a disposable stick, enabling direct sampling within the buccal cavity. An ad hoc designed 3D-printed attachment is used to ensure accurate positioning and alignment of the stick, as well as isolation from external lighting conditions. A custom-developed smartphone application captures and automatically processes the image of the sensing membrane, providing the salivary pH results. After optimizing the cocktail composition, the developed sensors demonstrated the capacity to determine pH within a range of 5.4 to 8.1 with a remarkable precision of 0.6%, achieving a very short analysis time of just 1 min. A stability study conducted on the sensing membranes revealed a lifetime of 50 days. To validate the performance of our analytical device, we compared its results against those obtained from a calibrated pH-meter, using a group of individuals. The device exhibited an average error of 2.4% when compared with the pH-meter results, confirming its reliability and accuracy.

Using a cotton thread-based colorimetric sensor modified by carboxymethylcellulose and cuprizone with smartphone detection for quantification of copper.

In the present work, we report the development of a novel cotton thread-based colorimetric sensor modified by carboxymethylcellulose (CMC) and cuprizone (CPZ) with smartphone detection and its application for the quantitative determination of cupric ions in water and cachaça. The cotton thread/smartphone detection-based colorimetric method is an easily affordable, low-cost technique which allows one to perform real-time and on-field determination analyses, especially with limited financial resources. The method involves the complexation of Cu(II) with CPZ, which causes a change in the coloration of the cotton thread from a shade of white to blue in the detection zone of the colorimetric sensor. The immobilization of CPZ on CMC in the cotton thread leads to the pre-concentration of Cu(II) via a complexation mechanism with colorimetric reaction. The application of the colorimetric sensor allows the quantification of copper in the range from 1 to 12 mg L-1, with a low limit of detection of 0.21 mg L-1. In addition, the recovery assays conducted in samples of water and cachaça resulted in recovery percentages ranging from 84.9% to 107%, which is indicative of a precise method. To validate the precision of the proposed colorimetric method, the values obtained from the quantification analysis were compared with those of the flame atomic absorption spectrometry and a good agreement at the 95% confidence level was obtained.

SARS-CoV-2 viral RNA detection using the novel CoVradar device associated with the CoVreader smartphone app.

The COVID-19 pandemic has highlighted the need for innovative approaches to its diagnosis. Here we present CoVradar, a novel and simple colorimetric method that combines nucleic acid analysis with dynamic chemical labeling (DCL) technology and the Spin-Tube device to detect SARS-CoV-2 RNA in saliva samples. The assay includes a fragmentation step to increase the number of RNA templates for analysis, using abasic peptide nucleic acid probes (DGL probes) immobilized to nylon membranes in a specific dot pattern to capture RNA fragments. Duplexes are formed by labeling complementary RNA fragments with biotinylated SMART bases, which act as templates for DCL. Signals are generated by recognizing biotin with streptavidin alkaline phosphatase and incubating with a chromogenic substrate to produce a blue precipitate. CoVradar results are analysed by CoVreader, a smartphone-based image processing system that can display and interpret the blotch pattern. CoVradar and CoVreader provide a unique molecular assay capable of detecting SARS-CoV-2 viral RNA without the need for extraction, preamplification, or pre-labeling steps, offering advantages in terms of time (∼3 h/test), cost (∼€1/test manufacturing cost) and simplicity (does not require large equipment). This solution is also promising for developing assays for other infectious diseases.

Using a biphasic system and digital imaging analysis with chemometric tools for simultaneous determination of Cu2+ and furfural in cachaça.

The present study reports, for the first time, the development and application of a highly efficient method based on digital imaging analysis for the simultaneous determination of Cu2+ and furfural in cachaça samples using a two-phase system and chemometrics tools. Furfural reacts with aniline in an acidic medium to form a Schiff base, which exhibits a pink color. On the other hand, Cu2+ reacts with cuprizone in a basic medium to form a blue complex. The two reactions were performed on a porcelain plate, and a smartphone was used to capture the colorimetric images. Partial least squares (PLS) regression was used to construct the prediction models for Cu2+ and furfural contents in cachaça samples. After finding the best PLS models, the ordered predictor selection (OPS) analysis was performed in order to select the most predictive variables. The method developed was found to be effective in estimating the amounts of Cu2+ and furfural in cachaça samples, with a mean absolute error of 0.2 mg L-1 for the Cu2+ model, and 0.3 mg per 100 mL of anhydrous alcohol for the furfural model. The method proposed in this study is simple and straightforward; it does not require complex technical knowledge and can be used by the producers themselves in the cachaça manufacturing process.

Near-Field Communication Tag for Colorimetric Glutathione Determination with a Paper-Based Microfluidic Device.

Here, we propose a microfluidic paper-based analytical device (µPAD) implemented with a near-field communication (NFC) tag as a portable, simple and fast colorimetric method for glutathione (GSH) determination. The proposed method was based on the fact that Ag+ could oxidize 3,3',5,5'-tetramethylbenzidine (TMB) into oxidized blue TMB. Thus, the presence of GSH could cause the reduction of oxidized TMB, which resulted in a blue color fading. Based on this finding, we developed a method for the colorimetric determination of GSH using a smartphone. A µPAD implemented with the NFC tag allowed the harvesting of energy from a smartphone to activate the LED that allows the capture of a photograph of the µPAD by the smartphone. The integration between electronic interfaces into the hardware of digital image capture served as a means for quantitation. Importantly, this new method shows a low detection limit of 1.0 µM. Therefore, the most important features of this non-enzymatic method are high sensitivity and a simple, fast, portable and low-cost determination of GSH in just 20 min using a colorimetric signal.

Reversal of a Fluorescent Fluoride Chemosensor from Turn-Off to Turn-On Based on Aggregation Induced Emission Properties.

Here we present a new approach for the development of fluoride chemosensors taking advantage of aggregation induced emission (AIE) properties. Although AIE-based chemosensors have been described, they rely primarily on the analyte causing aggregation and hence fluorescence. We propose a new concept in the use of AIE for the development of fluorescent sensors. Our hypothesis is based on the fact that a turn-off chemosensor in solution can be transformed into turn-on in the solid state if the properties of ACQ and AIE are properly combined between the fluorescent molecules involved. To demonstrate this hypothesis, we have selected a fluorescent chemosensor for the fluoride anion with a conjugated structure of bis(styryl)pyrimidine that, while showing turn-off behavior in solution, becomes turn-on when it is brought to the solid state. We have also combined it with the advantages of a detection system based on the microfluidic paper-based analytical devices (µPAD). The system is fully characterized spectroscopically both in solution and in the solid state, and quantum mechanical calculations were performed to explain how the sensor works. The prepared device presents a high sensitivity, with no interference and with an LoD and LoQ that allow determination of fluoride concentrations in water 2 orders of magnitude below the maximum allowed by WHO.

Microanalysis based on paper device functionalized with cuprizone to determine Cu2+ in sugar cane spirits using a smartphone.

In this work we propose for the first time, a paper-based test strip to analyse Cu2+ content in sugar cane spirits, which, due to its simplicity, high portability and fast analytical response (3 min), can be easily applied to in situ analyses by producers. The test strip was developed aiming: i) identify qualitatively the Cu2+ content in sugar cane spirits, and, ii) determine quantitatively the Cu2+ content using a digital image method employing a smartphone. The paper-based test strip was functionalized with cuprizone and optimized through a Box-Behnken, an experimental design for obtaining the best reaction conditions. Based on qualitative method with naked eyes approach performed by six volunteers analyst untrained, the method present a percentage of accuracy of 93%. For the quantitative analysis, it was determined the metal content at a level of statistical agreement with the reference method, as well as it was obtained the dynamic linear range from 2 to 13 mg L-1 with limits of detection and quantification of 0.034 and 0.103 mg L-1, respectively. Furthermore, the quantitative method showed a reliable precision with an RSD of 4.3% (n = 10) and the recovery of Cu2+ ranged from 80 to 103.8%.

Synthesis of a thermoresponsive crosslinked MEO2MA polymer coating on microclusters of iron oxide nanoparticles.

Encapsulation of magnetic nanoparticles (MNPs) of iron (II, III) oxide (Fe3O4) with a thermopolymeric shell of a crosslinked poly(2-(2-methoxyethoxy)ethyl methacrylate) P(MEO2MA) is successfully developed. Magnetic aggregates of large size, around 150-200 nm are obtained during the functionalization of the iron oxide NPs with vinyl groups by using 3-butenoic acid in the presence of a water soluble azo-initiator and a surfactant, at 70 °C. These polymerizable groups provide a covalent attachment of the P(MEO2MA) shell on the surface of the MNPs while a crosslinked network is achieved by including tetraethylene glycol dimethacrylate in the precipitation polymerization synthesis. Temperature control is used to modulate the swelling-to-collapse transition volume until a maximum of around 21:1 ratio between the expanded: shrunk states (from 364 to 144 nm in diameter) between 9 and 49 °C. The hybrid Fe3O4@P(MEO2MA) microgel exhibits a lower critical solution temperature of 21.9 °C below the corresponding value for P(MEO2MA) (bulk, 26 °C). The MEO2MA coating performance in the hybrid microgel is characterized by dynamic light scattering and transmission electron microscopy. The content of preformed MNPs [up to 30.2 (wt%) vs. microgel] was established by thermogravimetric analysis while magnetic properties by vibrating sample magnetometry.

Carbon Dots as Sensing Layer for Printed Humidity and Temperature Sensors.

This work presents an innovative application of carbon dots (Cdots) nanoparticles as sensing layer for relative humidity detection. The developed sensor is based on interdigitated capacitive electrodes screen printed on a flexible transparent polyethylene terephthalate (PET) film. Cdots are deposited on top of these electrodes. An exhaustive characterization of the nanoparticles has been conducted along with the fabrication of the sensor structure. The accompanied experiments give all the sensibility to the Cdots, showing its dependence with temperature and exciting frequency. To the best of our knowledge, this work paves the path to the use of these kind of nanoparticles in printed flexible capacitive sensors aimed to be employed in the continuously expanding Internet of Things ecosystem.

A vinyl sulfone clicked carbon dot-engineered microfluidic paper-based analytical device for fluorometric determination of biothiols.

A microfluidic paper-based analytical device integrating carbon dot (CDs) is fabricated and used for a fluorometric off-on assay of biothiols. Vinyl sulfone (VS) click immobilization of carbon dots (CDs) on paper was accomplished by a one-pot simplified protocol that uses divinyl sulfone (DVS) as a homobifunctional reagent. This reagent mediated both the click oxa-Michael addition to the hydroxyl groups of cellulose and ulterior covalent grafting of the resulting VS paper to NH2-functionalized CDs by means of click aza-Michael addition. The resulting cellulose nanocomposite was used to engineer an inexpensive and robust microfluidic paper-based analytical device (µPAD) that is used for a reaction-based off-on fluorometric assay of biothiols (GSH, Cys, and Hcy). The intrinsic blue fluorescence of CDs (with excitation/emission maxima at 365/450 nm) is turned off via the heavy atom effect of an introduced iodo group. Fluorescence is turned on again due to the displacement of iodine by reaction with a biothiol. The increase in fluorescence is related to the concentration over a wide range (1 to 200 µM for GSH and 5-200 µM for Cys and Hcy, respectively), and the assay exhibits a low detection limit (0.3 µM for GSH and Cys and 0.4 µM for Hcy). The method allows for rapid screening and can also be used in combination with a digital camera readout. Graphical abstract Schematic representation of a µPAD based on click immobilized carbon dots and used for a reaction-based fluorometric off-on assay of biothiols. The intrinsic blue fluorescence of carbon dots is turned off via the heavy atom effect of an introduced iodo group and turned on by the displacement of this atom by reaction with a biothiol.

Portable Instrument for Hemoglobin Determination Using Room-Temperature Phosphorescent Carbon Dots.

A portable reconfigurable platform for hemoglobin determination based on inner filter quenching of room-temperature phosphorescent carbon dots (CDs) in the presence of H2O2 is described. The electronic setup consists of a light-emitting diode (LED) as the carbon dot optical exciter and a photodiode as a light-to-current converter integrated in the same instrument. The reconfigurable feature provides adaptability to use the platform as an analytical probe for CDs coming from different batches with some variations in luminescence characteristics. The variables of the reaction were optimized, such as pH, concentration of reagents, and response time; as well as the variables of the portable device, such as LED voltage, photodiode sensitivity, and adjustment of the measuring range by a reconfigurable electronic system. The portable device allowed the determination of hemoglobin with good sensitivity, with a detection limit of 6.2 nM and range up to 125 nM.

Smartphone based meat freshness detection.

In this work, a freshness colorimetric sensor has been integrated with pork meat packages. The sensor tracks rising CO2 levels in the package associated with meat spoilage, as CO2 levels increase with bacterial population. The color of the sensor changes depending on the quantity of bacteria present, therefore it can be correlated with the freshness of meat, in this case pork loin. Detection is achieved by a simple photograph using a smartphone, and analyzing the grey scale from the RGB space color with a custom made app. Only 2 µL of the cocktail (all components are nontoxic) is needed to prepare the sensor, which have been integrated inside meat packages using a variety of support materials prior to sealing. The Smartphone measurements have been validated using a reference method (Checkpoint Analyzer) and the results suggest it can provide the basis for a quick test of the quality of the packaged pork.

Carbon dots-inspired fluorescent cyclodextrins: competitive supramolecular "off-on" (bio)sensors.

Chromophore-appended cyclodextrins combine the supramolecular loading capabilities of cyclodextrins (CDs) with the optical properties of the affixed chromophores. Among fluorescent materials, carbon dots (CNDs) are attractive and the feasibility of CND-appended CDs as sensors has been demonstrated by different authors. However, CNDs are intrinsically heterogeneous materials and their ulterior functionalization yields hybrid composites that are not well defined in terms of structure and composition. Inspired by the fluorescence properties of 5-oxo-1,2,3,5-tetrahydroimidazo[1,2-a]pyridine-7-carboxylic acid (IPCA), the most paradigmatic of the molecular fluorophores detected in CNDs, herein we report two highly efficient synthetic chemical strategies for the preparation of IPCA-appended CDs that behave as CND-based CD "turn off-on" biosensors suitable for the analysis of cholesterol and ß-galactosidase activity. We have deconstructed the CND-CD systems to demonstrate that (i) the role of CNDs is limited to acting as a support for the molecular fluorophores produced during their synthesis and (ii) the molecular fluorophores suffice for the determination of the enzymatic activity based on the quenching by p-nitrophenol as a sacrificial quencher.

General-purpose passive wireless point-of-care platform based on smartphone.

A versatile, compact and low-cost analytical platform has been designed, tested and validated to be used in the point-of-care settings. This passive measurement system is powered and complemented by a standard smartphone including a programmed application for measurement configuration and data processing as well as wireless results sharing. Electrochemical and electrochemiluminescence analytical techniques can be configured and realized by this platform that employs standard screen-printed electrodes for the sample managing and off-the-shelf electronic components. The power, electrical and optical signal processing have been studied in depth. The system can harvest energy up to 22.5 mW, set up a voltage in the range of ±1.15 V, and measure potentials in a range of 600 mV with an uncertainty of 1 mV, and current from 2 µA to 0.75 mA with a resolution of 1.1 µA. Moreover, standard tests have been performed to the platform consisting of amperometric, potentiometric, cyclic voltammetry and electrochemiluminescent analytical techniques, showing excellent agreement with a reference instrument. Finally, our design has also been applied to glucose, pH and H2O2 determinations, providing the full analytical parameters which are in very good agreement with the reference instrument results. Ranges (0.065-0.75 M, 0.62-100 mM and 3-9 pH units for glucose, H2O2 and pH, respectively) and limits of detection (0.024 M and 0.03 mM for glucose and H2O2, respectively) make this low-cost platform (

Acid anhydride coated carbon nanodots: activated platforms for engineering clicked (bio)nanoconstructs.

Activated carbon nanodots functionalized with acid anhydride groups (AA-CNDs) are prepared by one-pot water-free green thermolysis of citric acid. As a proof of concept of their capabilities as appealing and versatile platforms for accessing engineering nanoconstructs, the as-prepared AA-CNDs have been reacted to yield clickable CNDs. Their click bioconjugation with relevant recognizable complementary clickable sugars has led to multivalent CND-based glyconanoparticles that are non-toxic and biorecognizable. The accessibility and intrinsic reactivity of AA-CNDs expand the current toolbox of covalent surface grafting methodologies and provide a wide range of potential applications for engineering (bio)nanoconstructs.

Real time monitoring of glucose in whole blood by smartphone.

A combined thread-paper microfluidic device (µTPAD) is presented for the determination of glucose in blood. The device is designed to include all the analytical operations needed: red blood cell separation, conditioning, enzymatic recognition, and colorimetric transduction. The signal is captured with a smartphone or tablet working in video mode and processed by custom Android-based software in real-time. The automatic detection of the region of interest on the thread allows for the use of either initial rate or equilibrium signal as analytical parameters. The time needed for analysis is 12 s using initial rate, and 100 s using the equilibrium measurement with a LOD of 48 µM and 12 µM, respectively, and a precision around 7%. The µTPAD allows a rapid determination of glucose in real samples using only 3 µL of whole blood.

Ionophore-Based Optical Sensor for Urine Creatinine Determination.

Creatinine is a metabolite present in urine, and its concentration is used to diagnose and monitor kidney performance. For that reason, the development of new sensors to analyze this metabolite and obtain accurate results in a short period of time is necessary. An optical disposable sensor for monitoring creatinine levels in urine is described. The system, based on a new aryl-substituted calix[4]pyrrole synthetic receptor, has an unusual coextraction scheme. Due to the low p Ka values of creatininium (p Ka 4.8), a careful selection of a lipophilic pH indicator that works in acid medium is required. The sensor components were optimized, and the new sensor displays a good response time to creatinine (approximately 3 min) over a wide dynamic range (from 1 × 10-5 to 1 × 10-2 M). Moreover, the optical selectivity coefficients obtained for creatinine over common cations present in urine meet the requirements for real sample measurements. With a good sensor-to-sensor reproducibility (RSD, 5.1-6.9% in the middle of the range), this method provides a simple, quick, cost-effective, and selective alternative to the conventional methodology based on Jaffé's reaction.

Microfluidic paper-based device for colorimetric determination of glucose based on a metal-organic framework acting as peroxidase mimetic.

This work presents a microfluidic paper-based analytical device (µPAD) for glucose determination using a supported metal-organic framework (MOF) acting as a peroxidase mimic. The catalytic action of glucose oxidase (GOx) on glucose causes the formation of H2O2, and the MOF causes the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) by H2O2 to form a blue-green product with an absorption peak at 650 nm in the detection zone. A digital camera and the iOS feature of a smartphone are used for the quantitation of glucose with the S coordinate of the HSV color space as the analytical parameter. Different factors such as the concentration of TMB, GOx and MOF, pH and buffer, sample volume, reaction time and reagent position in the µPAD were optimized. Under optimal conditions, the value for the S coordinate increases linearly up to 150 µmol·L-1 glucose concentrations, with a 2.5 µmol·L-1 detection limit. The µPAD remains stable for 21 days under conventional storage conditions. Such an enzyme mimetic-based assay to glucose determination using Fe-MIL-101 MOF implemented in a microfluidic paper-based device possesses advantages over enzyme-based assays in terms of costs, durability and stability compared to other existing glucose determination methods. The procedure was applied to the determination of glucose in (spiked) serum and urine. Graphical abstract Schematic representation of microfluidic paper-based analytical device using metal-organic framework as a peroxidase mimic for colorimetric glucose detection with digital camera or smartphone and iOS app readout.

Tetrazine-based chemistry for nitrite determination in a paper microfluidic device.

We present a new chemistry to determine nitrites implemented in a microfluidic paper-based analytical device (µPAD). The device is fabricated in cellulose paper with a sample reception area and three replicate detection areas with recognition chemistry immobilized by adsorption. The method involves the use of nitrite in an acid medium reaction to generate nitrous acid, which produces the oxidation of s-dihydrotetrazine: 1,2-dihydro-3,6-bis(3,5-dimethyl-1H-pyrazol-1-yl)-1,2,4,5-tetrazine (DHBPTz), which change the detection zone from colorless to pink. We used a digital camera and smartphone for the quantitative analysis of nitrite with the color coordinate S of the HSV color space as the analytical parameter. Parameters such as concentration and volume of s-dihydrotetrazine, pH, sample volume and reaction time were studied. The detection limit for this method is 1.30µM nitrite. To estimate the selectivity of the method an interference study of common ions in water samples was performed. The procedure was applied to natural water and compared with reference procedures.