scholarly journals Smartphone-Based Chemiluminescent Origami µPAD for the Rapid Assessment of Glucose Blood Levels

Biosensors ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 381
Author(s):  
Donato Calabria ◽  
Martina Zangheri ◽  
Ilaria Trozzi ◽  
Elisa Lazzarini ◽  
Andrea Pace ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Rapid Assessment ◽  
Blood Levels ◽  
Blood Samples ◽  
3D Printed ◽  
Diagnostic Applications ◽  
Catalyzed Oxidation

Microfluidic paper analytical devices (µPADs) represent one of the most appealing trends in the development of simple and inexpensive analytical systems for diagnostic applications at the point of care (POC). Herein, we describe a smartphone-based origami µPAD for the quantitative determination of glucose in blood samples based on the glucose oxidase-catalyzed oxidation of glucose leading to hydrogen peroxide, which is then detected by means of the luminol/hexacyanoferrate(III) chemiluminescent (CL) system. By exploiting the foldable µPAD format, a two-step analytical procedure has been implemented. First, the diluted blood sample was added, and hydrogen peroxide was accumulated, then the biosensor was folded, and a transport buffer was added to bring hydrogen peroxide in contact with CL reagents, thus promoting the CL reaction. To enable POC applicability, the reagents required for the assay were preloaded in the µPAD so that no chemicals handling was required, and a 3D-printed portable device was developed for measuring the CL emission using the smartphone’s CMOS camera. The µPAD was stable for 30-day storage at room temperature and the assay, displaying a limit of detection of 10 µmol L−1, proved able to identify both hypoglycemic and hyperglycemic blood samples in less than 20 min.

scholarly journals Direct and rapid measurement of hydrogen peroxide in human blood using a microfluidic device

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
R. Gaikwad ◽  
P. R. Thangaraj ◽  
A. K. Sen
Keyword(s):  
Hydrogen Peroxide ◽  
Microfluidic Device ◽  
Human Blood ◽  
Blood Cells ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Time Interval ◽  
Fluorescence Measurement ◽  
Rapid Measurement ◽  
Automated Method

AbstractThe levels of hydrogen peroxide ($${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 ) in human blood is of great relevance as it has emerged as an important signalling molecule in a variety of disease states. Fast and reliable measurement of $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 levels in the blood, however, continues to remain a challenge. Herein we report an automated method employing a microfluidic device for direct and rapid measurement of $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 in human blood based on laser-induced fluorescence measurement. Our study delineates the critical factors that affect measurement accuracy—we found blood cells and soluble proteins significantly alter the native $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 levels in the time interval between sample withdrawal and detection. We show that separation of blood cells and subsequent dilution of the plasma with a buffer at a ratio of 1:6 inhibits the above effect, leading to reliable measurements. We demonstrate rapid measurement of $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 in plasma in the concentration range of 0–49 µM, offering a limit of detection of 0.05 µM, a sensitivity of 0.60 µM−1, and detection time of 15 min; the device is amenable to the real-time measurement of $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 in the patient’s blood. Using the linear correlation obtained with known quantities of $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 , the endogenous $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 concentration in the blood of healthy individuals is found to be in the range of 0.8–6 µM. The availability of this device at the point of care will have relevance in understanding the role of $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 in health and disease.

scholarly journals A Smartphone Camera Colorimetric Assay of Acetylcholinesterase and Butyrylcholinesterase Activity

Sensors ◽  
2021 ◽  
Vol 21 (5) ◽  
pp. 1796
Author(s):  
Miroslav Pohanka ◽  
Jitka Zakova
Keyword(s):  
Limit Of Detection ◽  
Colorimetric Assay ◽  
Point Of Care ◽  
Specific Treatment ◽  
Point Of Care Testing ◽  
Analytical Instruments ◽  
Colorimetric Test ◽  
3D Printed ◽  
Butyrylcholinesterase Activity ◽  
Additional Education

Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) can serve as biochemical markers of various pathologies like liver disfunction and poisonings by nerve agents. Ellman’s assay is the standard spectrophotometric method to measure cholinesterase activity in clinical laboratories. The authors present a new colorimetric test to assess AChE and BChE activity in biological samples using chromogenic reagents, treated 3D-printed measuring pads and a smartphone camera as a signal detector. Multiwell pads treated with reagent substrates 2,6-dichlorophenolindophenyl acetate, indoxylacetate, ethoxyresorufin and methoxyresorufin were prepared and tested for AChE and BChE. In the experiments, 3D-printed pads containing indoxylacetate as a chromogenic substrate were optimal for analytical purposes. The best results were achieved using the red (R) channel, where the limit of detection was 4.05 µkat/mL for BChE and 4.38 µkat/mL for AChE using a 40 µL sample and a 60 min assay. The major advantage of this method is its overall simplicity, as samples are applied directly without any specific treatment or added reagents. The assay was also validated to the standard Ellman’s assay using human plasma samples. In conclusion, this smartphone camera-based colorimetric assay appears to have practical applicability and to be a suitable method for point-of-care testing because it does not require specific manipulation, additional education of staff or use of sophisticated analytical instruments.

scholarly journals A simple spectrophotometric method for the determination of methyldopa using p-chloranil in the presence of hydrogen peroxide

2008 ◽  
Vol 33 (3) ◽  
pp. 7-12 ◽  
Author(s):  
M. A. Gotardo ◽  
L. S. Lima ◽  
R. Sequinel ◽  
J. L. Rufino ◽  
L. Pezza ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Spectrophotometric Method ◽  
Limit Of Detection ◽  
Pharmaceutical Formulations ◽  
Limit Of Quantification ◽  
Coefficients Of Variation ◽  
The Common ◽  
Interday Precision ◽  
Sensitive Spectrophotometric Method

A simple, rapid and sensitive spectrophotometric method has been developed for the determination of methyldopa in pharmaceutical formulations. The method is based on the reaction between tetrachloro-p-benzoquinone (p-chloranil) and methyldopa, accelerated by hydrogen peroxide (H2O2), producing a violet-red compound (λmax = 535 nm) at ambient temperature (25.0 ± 0.2 ºC). Experimental design methodologies were used to optimize the measurement conditions. Beer's law is obeyed in a concentration range from 2.10 x 10-4 to 2.48 x 10-3 mol L-1 (r = 0.9997). The limit of detection was 7.55 x 10-6 mol L-1 and the limit of quantification was 2.52 x 10-5 mol L-1. The intraday precision and interday precision were studied for 10 replicate analyses of 1.59 x 10-3 mol L-1 methyldopa solution and the respective coefficients of variation were 0.7 and 1.1 %. The proposed method was successfully applied to the determination of methyldopa in commercial brands of pharmaceuticals. No interferences were observed from the common excipients in the formulations. The results obtained by the proposed method were favorably compared with those given by the Brazilian Pharmacopoeia procedure at 95 % confidence level.

scholarly journals Kinetic spectrophotometric determination of Bi(III) based on its catalytic effect on the oxidation of phenylfluorone by hydrogen peroxide

2009 ◽  
Vol 74 (8-9) ◽  
pp. 977-984
Author(s):  
Sofija Rancic ◽  
Snezana Nikolic-Mandic
Keyword(s):  
Hydrogen Peroxide ◽  
Spectrophotometric Determination ◽  
Reaction Rate ◽  
Catalytic Effect ◽  
Limit Of Detection ◽  
Kinetic Method ◽  
Limit Of Quantification ◽  
Kinetic Spectrophotometric ◽  
Good Agreement

A new reaction was suggested and a new kinetic method was elaborated for determination of Bi(III) in solution, based on its catalytic effect on the oxidation of phenyl-fluorone (PF) by hydrogen peroxide in ammonia buffer. By application of spectrophotometric technique, a limit of quantification (LQ) of 128 ng cm-3 was reached, and the limit of detection (LD) of 37 ng cm-3 was obtained, where LQ was defined as the ratio signal: noise = 10:1 and LD was defined as signal 3:1 against the blank. The RSD value was found to be in the range 2.8-4.8 % for the investigated concentration range of Bi(III). The influence of some ions upon the reaction rate was tested. The method was confirmed by determining Bi(III) in a stomach ulcer drug ('Bicit HP', Hemofarm A.D.). The obtained results were compared to those obtained by AAS and good agreement of results was obtained.

scholarly journals Whole Blood Immunoassay Based on Centrifugal Bead Sedimentation

2011 ◽  
Vol 57 (5) ◽  
pp. 753-761 ◽  
Author(s):  
Ulrich Y Schaff ◽  
Greg J Sommer
Keyword(s):  
Whole Blood ◽  
Single Channel ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Low Cost ◽  
Microfluidic System ◽  
Blood Samples ◽  
Reactive Protein ◽  
Immunoassay Technique ◽  
Microfluidic Immunoassay

BACKGROUND Centrifugal “lab on a disk” microfluidics is a promising avenue for developing portable, low-cost, automated immunoassays. However, the necessity of incorporating multiple wash steps results in complicated designs that increase the time and sample/reagent volumes needed to run assays and raises the probability of errors. We present proof of principle for a disk-based microfluidic immunoassay technique that processes blood samples without conventional wash steps. METHODS Microfluidic disks were fabricated from layers of patterned, double-sided tape and polymer sheets. Sample was mixed on-disk with assay capture beads and labeling antibodies. Following incubation, the assay beads were physically separated from the blood cells, plasma, and unbound label by centrifugation through a density medium. A signal-laden pellet formed at the periphery of the disk was analyzed to quantify concentration of the target analyte. RESULTS To demonstrate this technique, the inflammation biomarkers C-reactive protein and interleukin-6 were measured from spiked mouse plasma and human whole blood samples. On-disk processing (mixing, labeling, and separation) facilitated direct assays on 1-μL samples with a 15-min sample-to-answer time, <100 pmol/L limit of detection, and 10% CV. We also used a unique single-channel multiplexing technique based on the sedimentation rate of different size or density bead populations. CONCLUSIONS This portable microfluidic system is a promising method for rapid, inexpensive, and automated detection of multiple analytes directly from a drop of blood in a point-of-care setting.

scholarly journals Rapid Detection of Bacillus anthracis Bloodstream Infections by Use of a Novel Assay in the GeneXpert System

2017 ◽  
Vol 55 (10) ◽  
pp. 2964-2971 ◽  
Author(s):  
Padmapriya P. Banada ◽  
Srinidhi Deshpande ◽  
Riccardo Russo ◽  
Eric Singleton ◽  
Darshini Shah ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Dynamic Range ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Severe Disease ◽  
Automated System ◽  
Bacterial Isolates ◽  
Sample Processing ◽  
Blood Samples ◽  
Content Type

ABSTRACT Bacillus anthracis is a tier 1 select agent with the potential to quickly cause severe disease. Rapid identification of this pathogen may accelerate treatment and reduce mortality in the event of a bioterrorism attack. We developed a rapid and sensitive assay to detect B. anthracis bacteremia using a system that is suitable for point-of-care testing. A filter-based cartridge that included both sample processing and PCR amplification functions was loaded with all reagents needed for sample processing and multiplex nested PCR. The assay limit of detection (LOD) and dynamic range were determined by spiking B. anthracis DNA into individual PCR mixtures and B. anthracis CFU into human blood. One-milliliter blood samples were added to the filter-based detection cartridge and tested for B. anthracis on a GeneXpert instrument. Assay specificity was determined by testing blood spiked with non-anthrax bacterial isolates or by testing blood samples drawn from patients with concurrent non- B. anthracis bacteremia or nonbacteremic controls. The assay LODs were 5 genome equivalents per reaction and 10 CFU/ml blood for both the B. anthracis Sterne and V1B strains. There was a 6-log 10 dynamic range. Assay specificity was 100% for tests of non- B. anthracis bacterial isolates and patient blood samples. Assay time was less than 90 min. This automated system suitable for point-of-care detection rapidly identifies B. anthracis directly from blood with high sensitivity. This assay might lead to early detection and more rapid therapy in the event of a bioterrorism attack.

scholarly journals Label-Free Colorimetric Detection of Urine Glucose Based on Color Fading Using Smartphone Ambient-Light Sensor

Chemosensors ◽  
2020 ◽  
Vol 8 (1) ◽  
pp. 10 ◽  
Author(s):  
Wang ◽  
Guo ◽  
Hu ◽  
Liang ◽  
Li ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Colorimetric Assay ◽  
Point Of Care ◽  
Colorimetric Detection ◽  
Diabetic Patients ◽  
Ambient Light ◽  
Label Free ◽  
Urine Glucose ◽  
Light Sensor

In this work, a label-free colorimetric assay was developed for the determination of urine glucose using smartphone ambient-light sensor (ALS). Using horseradish peroxidase—hydrogen peroxide—3,3′,5,5′-tetramethylbenzidine (HRP-H2O2-TMB) colored system, quantitative H2O2 was added to samples to-be-determined for deepest color. The presence of glucose oxidase in urine led to the formation of H2O2 and the reduction of TMBred. As a result of this, the color of the urine faded and the solution changed from deep blue to light blue. We measured the illuminance of the transmitted light by a smartphone ambient light sensor, and thereby color changes were used to calculate the content of urine glucose. After method validation, this colorimetric assay was practically applied for the determination of urine samples from diabetic patients. Good linearity was obtained in the range of 0.039–10.000 mg/mL (R2 = 0.998), and a limit of detection was 0.005 mg/mL. Our method was had high accuracy, sensitivity, simplicity, rapidity, and visualization, providing a new sensor to be potentially applicable for point-of-care detection of urine glucose.

Application of Homogeneous Liquid–Liquid Microextraction With Switchable Hydrophilicity Solvents to the Determination of MDMA, MDA and NBOMes in Postmortem Blood Samples

2021 ◽  
Author(s):  
Camila Scheid ◽  
Sarah Eller ◽  
Anderson Luiz Oenning ◽  
Eduardo Carasek ◽  
Josias Merib ◽  
...  
Keyword(s):  
Illicit Drugs ◽  
Matrix Effects ◽  
Limit Of Detection ◽  
Blood Samples ◽  
Synthetic Drugs ◽  
Preparation Conditions ◽  
Limit Of Quantitation ◽  
Postmortem Blood ◽  
Liquid Microextraction

Abstract Synthetic drugs for recreational purposes are in constant evolution, and their consumption promotes a significant increase in intoxication cases, resulting in damaging public health. The development of analytical methodologies to confirm the consumption of illicit drugs in biological matrices is required for the control of these substances. This work exploited the development of an extraction method based on homogenous liquid–liquid microextraction with switchable hydrophilicity solvent (SHS) as extraction phase for the determination of the synthetic drugs 3,4-methylenedioxymethamphetamine, 3,4-methylenedioxyamphetamine and N-methoxybenzyl-methoxyphenylethylamine derivates (25B, 25C and 25I) in postmortem blood, followed by liquid chromatography coupled to mass spectrometry in tandem. The optimized sample preparation conditions consisted of using 250 µL of ZnSO4 10% and 50 µL of NaOH 1 mol/L in the protein precipitation step; N,N-dimethylcyclohexylamine was used as SHS, 650 μL of a mixture of SHS:HCl 6 mol/L (1:1 v/v), 500 μL of whole blood, 500 μL of NaOH 10 mol/L and 1 min of extraction time. The proposed method was validated, providing determination coefficients higher than 0.99 for all analytes; limit of detection and limit of quantitation ranged from 0.1 to 10 ng/mL; intra-run precision from 2.16% to 9.19%; inter-run precision from 2.39% to 9.59%; bias from 93.57% to 115.71% and matrix effects from 28.94% to 51.54%. The developed method was successfully applied to four authentic postmortem blood samples from synthetic drugs users, and it was found to be reliable with good selectivity.

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