Thiago M. G. Cardoso

A handheld stamping process to fabricate microfluidic paper-based analytical devices with chemically modified surface for clinical assays

This paper describes the development and use of a handheld and lightweight stamp for the production of microfluidic paper-based analytical devices (μPADs). We also chemically modified the paper surface for improved colorimetric measurements. The design of the microfluidic structure has been patterned in a stamp, machined in stainless steel. Prior to stamping, the paper surface was oxidized to promote the conversion of hydroxyl into aldehyde groups, which were then chemically activated for covalent coupling of enzymes. Then, a filter paper sheet was impregnated with paraffin and sandwiched with a native paper (n-paper) sheet, previously oxidized. The metal stamp was preheated at 150 °C and then brought in contact with the paraffined paper (p-paper) to enable the thermal transfer of the paraffin to the n-paper, thus forming the hydrophobic barriers under the application of a pressure of ca. 0.1 MPa for 2 s. The channel and barrier widths measured in 50 independent μPADs exhibited values of 2.6 ± 0.1 and 1.4 ± 0.1 mm, respectively. The chemical modification for covalent coupling of enzymes on the paper surface also led to improvements in the colour uniformity generated inside the sensing area, a known bottleneck in this technology. The relative standard deviation (RSD) values for glucose and uric acid (UA) assays decreased from 40 to 10% and from 20 to 8%, respectively. Bioassays related to the detection of glucose, UA, bovine serum albumin (BSA), and nitrite were successfully performed in concentration ranges useful for clinical assays. The semi-quantitative analysis of all four analytes in artificial urine samples revealed an error smaller than 4%. The disposability of μPADs, the low instrumental requirements of the stamp-based fabrication, and the improved colour uniformity enable the use of the proposed devices for the point-of-care diagnostics or in limited resources settlements.

Colorimetric determination of nitrite in clinical, food and environmental samples using microfluidic devices stamped in paper platforms

This study reports the use of microfluidic paper-based analytical devices (μPADs) associated with colorimetric detection for the determination of nitrite in clinical, food and environmental samples. μPADs were fabricated by a simple and fast stamping process in a geometry containing eight circular detection zones and one central zone to sample inlet interconnected by microfluidic channels. The colorimetric determination of nitrite was performed through the modified Griess reaction. Detection zones were spotted with a 0.75 μL aliquot of a solution containing 50 mM sulfanilamide, 1.2 M hydrochloric acid and 4 mM N-(1-naphthyl)ethylenediamine. The monitoring of the background colorimetric response revealed good stability over 12 h for devices stored in the absence of light. After the addition of standard or real samples, the resulting images were captured with a scanner, converted to a color scale and analyzed in the magenta channel. The analytical sensitivity and the limit of detection achieved after a preconcentration stage were 0.56 (AU μM−1) and 5.6 μM, respectively. The preconcentration provided an enrichment factor of ca. 3.2 times. The concentration levels of nitrite were successfully determined in saliva, preservative water, ham, sausage and river water samples. The concentration levels attained for each sample using μPADs were compared to the values found by spectrophotometry and there was no significant difference from one another at a confidence level of 95%.

A fully disposable paper-based electrophoresis microchip with integrated pencil-drawn electrodes for contactless conductivity detection

We describe for the first time the fabrication of a paper-based microchip electrophoresis (pME) device with integrated hand-drawn pencil electrodes to perform capacitively coupled contactless conductivity detection (C4D). This low-cost device (less than

Versatile fabrication of paper-based microfluidic devices with high chemical resistance using scholar glue and magnetic masks

0.10) revealed great capability to dissipate heat, good injection-to-injection repeatability, and ease of attaching pencil-drawn electrodes on the separation channel. The feasibility of the proposed pME-C4D device was successfully demonstrated with the separation of bovine serum albumin and creatinine within 150 s with baseline resolution. The limits of detection values for albumin and creatinine were 20 and 35 μmol L−1, respectively. These biomolecules present clinical relevance as evidence of kidney failure. The proposed pME-C4D offers great potential to be explored in the diagnosis of diabetes mellitus and heart disease.

Determination of Ascorbic Acid in Commercial Tablets Using Pencil Drawn Electrochemical Paper-based Analytical Devices

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

Portable analytical platforms for forensic chemistry: A review

This study describes the use of electrochemical paper-based analytical devices (ePADs) drawn with graphite pencil for the determination of ascorbic acid (AA) in commercial tablets. ePADs were fabricated using vegetal paper and graphite pencil. First, the three-electrode electrochemical cell drawn using a graphical software and toner lines were laser printed on the vegetal paper surface to delimit the electrode areas. Then, the electrode regions were manually painted with graphite pencil. Afterwards, the pseudo-reference electrode was defined with the deposition of silver ink over the graphite surface. Cyclic voltammetry and square wave voltammetry (SWV) experiments were performed to optimize the electroanalytical parameters as well as to quantitatively determine the AA concentration in two commercial tables. ePADs exhibited linear behavior for a concentration range between 0.5 and 3.0 mmol L-1. The achieved limit of detection and sensitivity were 70 μmol L-1 and 0.47 μA/mmol L-1, respectively. The AA concentration levels found by SWV experiments in both CenevitTM and Energil CTM were 2.80 ± 0.02 and 3.10 ± 0.01 mmol L-1, respectively. The accuracy of the proposed devices was investigated through recovery experiments in three concentration levels and it presented values between 95 and 115%.

Enhanced Performance of Colorimetric Biosensing on Paper Microfluidic Platforms Through Chemical Modification and Incorporation of Nanoparticles

This current review article focuses on recent contributions to on-site forensic investigations. Portable and potentially portable methods are presented and critically discussed about (bio)chemical trace analysis and studies performed outside the controlled laboratory environment to rapidly help in crime scene inquiries or forensic intelligence purposes. A wide range of approaches including electrochemical sensors, microchip electrophoresis, ambient ionization on portable mass spectrometers, handheld Raman and NIR instruments as well as and point-of-need devices, like paper-based platforms, for in-field analysis of latent evidences, controlled substances, drug screening, hazards, and others to assist in law enforcements and solving crime more efficiently are highlighted. The covered examples have successfully demonstrated the huge potential of portable devices for on-site applications. Future investigations should consider analytical validation to compete equality and even replace current gold standard methods.

Paper-based enzymatic reactors for batch injection analysis of glucose on 3D printed cell coupled with amperometric detection

This chapter describes two different methodologies used to improve the analytical performance of colorimetric paper-based biosensors. Microfluidic paper-based analytical devices (μPADs) have been produced by a stamping process and CO2 laser ablation and modified, respectively, through an oxidation step and incorporation of silica nanoparticles on the paper structure. Both methods are employed in order to overcome the largest problem associated with colorimetric detection, the heterogeneity of the color distribution in the detection zones. The modification steps are necessary to improve the interaction between the paper surface and the selected enzymes. The enhanced performance has ensured reliability for quantitative analysis of clinically relevant compounds.