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Link to original content: http://pubmed.ncbi.nlm.nih.gov/37366964/
Affordable Portable Platform for Classic Photometry and Low-Cost Determination of Cholinesterase Activity - PubMed Skip to main page content
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. 2023 May 31;13(6):599.
doi: 10.3390/bios13060599.

Affordable Portable Platform for Classic Photometry and Low-Cost Determination of Cholinesterase Activity

Ondřej Keresteš  1 , Miroslav Pohanka  1
Affiliations

Affordable Portable Platform for Classic Photometry and Low-Cost Determination of Cholinesterase Activity

Ondřej Keresteš et al. Biosensors (Basel). .

Abstract

Excessive use of pesticides could potentially harm the environment for a long time. The reason for this is that the banned pesticide is still likely to be used incorrectly. Carbofuran and other banned pesticides that remain in the environment may also have a negative effect on human beings. In order to provide a better chance for effective environmental screening, this thesis describes a prototype of a photometer tested with cholinesterase to potentially detect pesticides in the environment. The open-source portable photodetection platform uses a color-programmable red, green and blue light-emitting diode (RGB LED) as a light source and a TSL230R light frequency sensor. Acetylcholinesterase from Electrophorus electricus (AChE) with high similarity to human AChE was used for biorecognition. The Ellman method was selected as a standard method. Two analytical approaches were applied: (1) subtraction of the output values after a certain period of time and (2) comparison of the slope values of the linear trend. The optimal preincubation time for carbofuran with AChE was 7 min. The limits of detection for carbofuran were 6.3 nmol/L for the kinetic assay and 13.5 nmol/L for the endpoint assay. The paper demonstrates that the open alternative for commercial photometry is equivalent. The concept based on the OS3P/OS3P could be used as a large-scale screening system.

Keywords: affordable; cholinesterase; field-analysis; open-source; pesticide; photometry; point-of-care test.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Schematic of OS3P box with inner parts placement—(“A”) LED module holder. (“B”) Cover of parasitic light from the LED on the Arduino UNO R3 board. (“C”) Cuvette stabilizer. It is used to stabilize the standard position of the cuvette and prevent the cuvette from failing. (“D”) Hole for USB-B connector. (“E”) Cuvette hole with reinforced wall for better cuvette insertion.
Figure 2
Figure 2
(A) Schematic of an Open-source portable photodetection platform (OS3P). (B) Overview of the connected hardware.
Figure 3
Figure 3
Cutaway view of the OS3P measuring aperture after box assembly.
Figure 4
Figure 4
Emission aperture details. An optical fiber connected to Spectrovis Plus was used. The red, green and blue light-emitting diode (RGB LED) module was controlled by Arduino UNO and set up for maximum intensity.
Figure 5
Figure 5
Data acquisition procedure with Open-source portable photometric platform (OS3P). The analysis algorithm is shown and each of the consecutive steps is numbered. Created with BioRender.com.
Figure 6
Figure 6
(A) Emission spectra of the KS0032 RGB LED module. (B) Comparison of the response of the TSL230R to different light conditions. The error bars in B indicate the standard deviation for n = 10 (no light) and n = 30 (with the light source was turned on).
Figure 7
Figure 7
Phenol red calibration curve in the wavelength mode. Comparison between benchtop analyzer (A) and OS3P (B). Error bars indicate standard deviation for n = 3 (A); n = 10 (B).
Figure 8
Figure 8
Saturation curves of acetylcholinesterase. Measurement using OS3P. The error bars indicate the standard error for n = 3.
Figure 9
Figure 9
Real-time monitoring of cholinesterase activity. (A) Standard response of the enzyme assay. Evaluated part of the curves used to determine the kinetic parameters (B).
Figure 10
Figure 10
The optimal incubation time of carbofuran with AChE was measured using the Evolution 201 benchtop spectrophotometer to optimize the preincubation time of the inhibitor.
Figure 11
Figure 11
Calibration curves of carbofuran. The inverted slope values (A) and the difference in output frequency (B) were obtained and evaluated as separate markers of residual AChE activity.
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