Batch injection analysis towards auxiliary diagnosis of periodontal diseases based on indirect amperometric detection of salivary α-amylase on a cupric oxide electrode.

This study describes, for the first time, the use of a batch injection analysis system with amperometric detection (BIA-AD) to indirectly determine salivary α-amylase (sAA) levels in saliva samples for chronic periodontitis diagnosis. A chemical/thermal treatment was explored to generate a CuO film on a Cu electrode surface. This procedure offered good stability (RSD = 0.3%), good repeatability (RSD < 1.3%) and excellent reproducibility (RSD < 1.5%). The sAA concentration levels were determined based on the detection of maltose produced by enzymatic hydrolysis of starch. The analytical performance was investigated, and a linear correlation was observed for a maltose concentration range between 0.5 and 6.0 mmol L-1 with a correlation coefficient equal to 0.999. The analytical sensitivity and the limit of detection were 48.8 µA/(mmol L-1) and 0.05 mmol L-1, respectively. In addition, the proposed system provided an excellent analytical frequency (120 analysis h-1). The clinical feasibility of the proposed method was investigated by the determination of sAA levels in four saliva samples (two from healthy control persons (C1 and C2) and two from patients with chronic periodontitis (P1 and P2)). The accuracy provided by the BIA-AD system ranged from 93 to 98%. The sAA concentration levels achieved for each sample were compared to the values found by spectrophotometry and there was no statistically significant difference between them at a confidence level of 95%. Finally, the method reported herein emerges as a simple, low cost and promising tool for assisting periodontal diseases diagnosis.

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

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.

Paper-based microfluidic devices on the crime scene: A simple tool for rapid estimation of post-mortem interval using vitreous humour.

This paper describes for the first time the use of paper-based analytical devices at crime scenes to estimate the post-mortem interval (PMI), based on the colorimetric determination of Fe2+ in vitreous humour (VH) samples. Experimental parameters such as the paper substrate, the microzone diameter, the sample volume and the 1,10-phenanthroline (o-phen) concentration were optimised in order to ensure the best analytical performance. Grade 1 CHR paper, microzone with diameter of 5 mm, a sample volume of 4 µL and an o-phen concentration of 0.05 mol/L were chosen as the optimum experimental conditions. A good linear response was observed for a concentration range of Fe2+ between 2 and 10 mg/L and the calculated values for the limit of detection (LOD) and limit of quantification (LOQ) were 0.3 and 0.9 mg/L, respectively. The specificity of the Fe2+ colorimetric response was tested in the presence of the main interfering agents and no significant differences were found. After selecting the ideal experimental conditions, four HV samples were investigated on paper-based devices. The concentration levels of Fe2+ achieved for samples #1, #2, #3 and #4 were 0.5 ± 0.1, 0.7 ± 0.1, 1.2 ± 0.1 and 15.1 ± 0.1 mg/L, respectively. These values are in good agreement with those calculated by ICP-MS. It important to note that the concentration levels measured using both techniques are proportional to the PMI. The limitation of the proposed analytical device is that it is restricted to a PMI greater than 1 day. The capability of providing an immediate answer about the PMI on the crime scene without any sophisticated instrumentation is a great achievement in modern instrumentation for forensic chemistry. The strategy proposed in this study could be helpful in many criminal investigations.

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

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.

Highly sensitive colorimetric detection of glucose and uric acid in biological fluids using chitosan-modified paper microfluidic devices.

This paper describes the modification of microfluidic paper-based analytical devices (µPADs) with chitosan to improve the analytical performance of colorimetric measurements associated with enzymatic bioassays. Chitosan is a natural biopolymer extensively used to modify biosensing surfaces due to its capability of providing a suitable microenvironment for the direct electron transfer between an enzyme and a reactive surface. This hypothesis was investigated using glucose and uric acid (UA) colorimetric assays as model systems. The best colorimetric sensitivity for glucose and UA was achieved using a chromogenic solution composed of 4-aminoantipyrine and sodium 3,5-dichloro-2-hydroxy-benzenesulfonate (4-AAP/DHBS), which provided a linear response for a concentration range between 0.1 and 1.0 mM. Glucose and UA were successfully determined in artificial serum samples with accuracies between 87 and 114%. The limits of detection (LODs) found for glucose and UA assays were 23 and 37 µM, respectively. The enhanced analytical performance of chitosan-modified µPADs allowed the colorimetric detection of glucose in tear samples from four nondiabetic patients. The achieved concentration levels ranged from 130 to 380 µM. The modified µPADs offered analytical reliability and accuracy as well as no statistical difference from the values achieved through a reference method. Based on the presented results, the proposed µPAD can be a powerful alternative tool for non-invasive glucose analysis.

Enhanced Analytical Performance of Paper Microfluidic Devices by Using Fe3O4 Nanoparticles, MWCNT, and Graphene Oxide.

Spheres, tubes, and planar-shaped nanomaterials as Fe3O4 nanoparticles (MNPs), multiwalled carbon nanotubes (MWCNT), and graphene oxide (GO) were used for the first time to treat microfluidic paper-based analytical devices (µPADs) and create a biocompatible layer with high catalytic surface. Once glucose measurements are critical for diabetes or glycosuria detection and monitoring, the analytical performance of the proposed devices was studied by using bienzymatic colorimetric detection of this carbohydrate. The limit of detection values achieved for glucose with µPADs treated with MNPs, MWCNT, and GO were 43, 62, and 18 µM, respectively. The paper surface modification solves problems associated with the lack of homogeneity on color measurements that compromise the sensitivity and detectability levels in clinical diagnosis.