Spectrophotomeric Assay of Aflatoxin B1 Using Acetylcholinesterase Immobilized on Standard Microplates

Pohanka, Miroslav

doi:10.1080/00032719.2012.757703

Cited by 20 publications

(7 citation statements)

References 24 publications

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“…It was reported for membrane-bound AChE (Korpela & Tahti, 1988 ) and the synergetic effect of organic solvents and pesticides (Turdean & Turdean, 2008 ). Our finding that the immobilized AChE is well stable once immobilized into a gelatin membrane is contradictory to our previous results (Pohanka, 2013 ). The immobilization of AChE into gelatin is probably better for stabilizing AChE than the previously used capturing into precipitate by albumin and glutaraldehyde.…”

Section: Resultscontrasting

confidence: 99%

Preparation and performance of a colorimetric biosensor using acetylcholinesterase and indoxylacetate for assay of nerve agents and drugs

Pohanka

Vlček

2014

Interdisciplinary Toxicology

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Different toxic compounds can target the cholinergic nervous system. Acetylcholinesterase (AChE; EC 3.1.1.7) is one of the most crucial components of the cholinergic nervous system and thus many of the toxins interact with this enzyme. As to inhibitors, nerve agents used as chemical warfare, some insecticides, and drugs influencing the cholinergic system are common examples of AChE inhibitors. Once inhibited by a neurotoxic compound, a serious cholinergic crisis can occur. On the other hand, sensitivity of AChE to the inhibition can be used for analytical purposes. In this study, a simple disposable biosensor with AChE as a recognition element was devised. AChE was immobilized onto a cellulose matrix and indoxylacetate was used as a chromogenic substrate. The enzyme reaction was assessed by the naked eye using arbitrary units and pyridostigmine, tacrine, paraoxon, carbofuran, soman and VX were assayed as selected inhibitors. A good stability of the biosensors was found, with no aging over a quarter of a year and minimal sensitivity to the interference of organic solvents. The limit of detection ranged from 10 to 100 nmol/L for the compounds tested with a sample volume of 40 µL.

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Section: Resultscontrasting

confidence: 99%

Preparation and performance of a colorimetric biosensor using acetylcholinesterase and indoxylacetate for assay of nerve agents and drugs

Pohanka

Vlček

2014

Interdisciplinary Toxicology

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“…An alternative method of determining aflatoxin is an inhibitory analysis using the enzyme biosensors based on acetylcholinesterase (АChЕ) [5][6][7][8]. The biosensor functioning is based on the following enzymatic reaction:…”

Section: Introductionmentioning

confidence: 99%

Inhibition of immobilized acetylcholinesterase by aflatoxin B1 in a potentiometric biosensor

et al. 2016

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To identify a type of inhibition of immobilized acetylcholinesterase by aflatoxin B1. Methods. A bioselective element of the potentiometric biosensor was created using acetylcholinesterase, which was covalently immobilized on the surface of the pH-FET sensor by glutaraldehyde crosslinking with bovine serum albumin. Results. Optimal conditions for the potentiometric biosensor operation such as pH-optimum of the enzyme action and its inhibition were defined. An apparent Michaelis constant, as well as a maximum initial reaction rate of immobilized acetylcholinesterase as a part of the biosensor were determined. The type of reversible inhibition of immobilized acetylcholinesterase by aflatoxin B1 in potentiometric biosensor was identified by using a new graphical "degree of inhibition" method and the obtained result was confirmed with one of the traditional methods, such as the Lineweaver-Burk plot. Conclusions. This study helps to understand the mechanisms of enzyme inhibition in biosensors and brings the biosensor implementation closer. K e y w o r d s: biosensor, immobilized acetylcholinesterase, type of inhibition, aflatoxin B1, potentiometric transducer.

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“…The mercapto groups are easily oxidized and the photocurrent response increases as a result of the effect of electron-hole pairs system. AFB 1 has a strong inhibitory effect on acetylcholinesterase and irreversibly noncompetitive inhibition by altering the site of action of acetylcholinesterase [18][19][20][21][22]. The reaction formula is as follows:…”

Section: Procedures For Electrochemical Biosensormentioning

confidence: 99%

Detection of AFB1 via TiO2 Nanotubes/Au Nanoparticles/Enzyme Photoelectrochemical Biosensor

Yuan

et al. 2018

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TiO 2 nanotubes/Au nanoparticles/enzyme photoelectrochemical biosensor is developed by the chemical bonding of acetylcholinesterase with Au nanoparticles-modified TiO 2 photoactive electrode, based on the inhibitory effect of aflatoxin B 1 on acetylcholinesterase activity. In this method, AuNPs were deposited on the surface of the electrode by potentiostatic deposition and the acetylcholinesterase was chemically crosslinked to the surface for determination of aflatoxin B 1 . Enzymatic hydrolysate is generated to capture the photogenerated holes of UV-sensitized TiO 2 nanotube arrays, causing magnification of the photoelectrochemical signal. The photoelectrochemical biosensor morphological and structural details were evaluated, applying different techniques, such as X-ray Diffraction (XRD) and Scanning Electron Microscopy (SEM). Aflatoxin B 1 competitively inhibits acetylcholinesterase, leading to a decrease in photocurrent that should have been increased. The detection performance of biosensors for different concentrations of AFB 1 is discussed. The linear response range of the biosensor is from 1-6 nM with detection limitation of 0.33 nM, the linear equation is I (µA) = −0.13C (nM) + 9.98 (µA), with a correlation coefficient of 0.988. This new biosensor could be used to detect Aflatoxin B 1 in foods.

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Spectrophotomeric Assay of Aflatoxin B1 Using Acetylcholinesterase Immobilized on Standard Microplates

Cited by 20 publications

References 24 publications

Preparation and performance of a colorimetric biosensor using acetylcholinesterase and indoxylacetate for assay of nerve agents and drugs

Preparation and performance of a colorimetric biosensor using acetylcholinesterase and indoxylacetate for assay of nerve agents and drugs

Inhibition of immobilized acetylcholinesterase by aflatoxin B1 in a potentiometric biosensor

Detection of AFB1 via TiO2 Nanotubes/Au Nanoparticles/Enzyme Photoelectrochemical Biosensor

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