Enzymatic Spectrophotometric Method for Aflatoxin B Detection Based on Acetylcholinesterase Inhibition
A new method for aflatoxin B (AFB) determination is proposed. The AFB determination is based on acetylcholinesterase (AChE) inhibition, and the AChE residual activity is determined using the colorimetric method (Ellman's method). Cholinesterases (ChEs) from various sources were tested using AFB...
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Veröffentlicht in: | Analytical chemistry (Washington) 2007-05, Vol.79 (9), p.3409-3415 |
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creator | Arduini, Fabiana Errico, Ilenia Amine, Aziz Micheli, Laura Palleschi, Giuseppe Moscone, Danila |
description | A new method for aflatoxin B (AFB) determination is proposed. The AFB determination is based on acetylcholinesterase (AChE) inhibition, and the AChE residual activity is determined using the colorimetric method (Ellman's method). Cholinesterases (ChEs) from various sources were tested using AFB1 as reference aflatoxin. AChE from electric eel has shown the highest sensitivity to AFB1, and it was chosen for the rest of the work. To select and optimize the analytical procedures, an investigation on the type of AChE inhibition by AFB1 was carried out. The AChE degree of inhibition by AFB1 was independent of the incubation time and the enzyme concentrations, showing the reversibility of the inhibition. This reversibility of the inhibition permits a rapid analysis of AFB1, requiring only 3 min. For the development of the AFB1 assay, the pH, the time of reaction, temperature, and substrate concentration were evaluated and optimized. The linear range of 10−60 ng mL-1 was determined. To evaluate the selectivity of this method, the cross-reactivity with other aflatoxins such as aflatoxin B2, aflatoxin G1, aflatoxin G2, and aflatoxin M1 was investigated. Finally, the suitability of the assay for AFB1 quantification in barley was evaluated. This study shows a new approach to detect aflatoxins based on enzyme inhibition and has advantages such as the ease of use, rapidity, and cost effectiveness. Thus, it could find a possible use as a screening method for this type of mycotoxins. |
doi_str_mv | 10.1021/ac061819j |
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The AFB determination is based on acetylcholinesterase (AChE) inhibition, and the AChE residual activity is determined using the colorimetric method (Ellman's method). Cholinesterases (ChEs) from various sources were tested using AFB1 as reference aflatoxin. AChE from electric eel has shown the highest sensitivity to AFB1, and it was chosen for the rest of the work. To select and optimize the analytical procedures, an investigation on the type of AChE inhibition by AFB1 was carried out. The AChE degree of inhibition by AFB1 was independent of the incubation time and the enzyme concentrations, showing the reversibility of the inhibition. This reversibility of the inhibition permits a rapid analysis of AFB1, requiring only 3 min. For the development of the AFB1 assay, the pH, the time of reaction, temperature, and substrate concentration were evaluated and optimized. The linear range of 10−60 ng mL-1 was determined. To evaluate the selectivity of this method, the cross-reactivity with other aflatoxins such as aflatoxin B2, aflatoxin G1, aflatoxin G2, and aflatoxin M1 was investigated. Finally, the suitability of the assay for AFB1 quantification in barley was evaluated. This study shows a new approach to detect aflatoxins based on enzyme inhibition and has advantages such as the ease of use, rapidity, and cost effectiveness. Thus, it could find a possible use as a screening method for this type of mycotoxins.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/ac061819j</identifier><identifier>PMID: 17408242</identifier><identifier>CODEN: ANCHAM</identifier><language>eng</language><publisher>Washington, DC: American Chemical Society</publisher><subject>Acetylcholinesterase - chemistry ; Acetylcholinesterase - drug effects ; Aflatoxin B1 - analysis ; Aflatoxin B1 - pharmacology ; Analytical chemistry ; Animals ; Chemistry ; Electrophorus ; Enzyme Activation - drug effects ; Exact sciences and technology ; Methanol - chemistry ; Methanol - pharmacology ; Molecular Structure ; Sensitivity and Specificity ; Spectrometric and optical methods ; Spectrophotometry - methods ; Structure-Activity Relationship ; Temperature</subject><ispartof>Analytical chemistry (Washington), 2007-05, Vol.79 (9), p.3409-3415</ispartof><rights>Copyright © 2007 American Chemical Society</rights><rights>2007 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a447t-ba9bcb0af800388ceba53a0643ad05b869ad1e6603f08199f951e84b1f8c79cd3</citedby><cites>FETCH-LOGICAL-a447t-ba9bcb0af800388ceba53a0643ad05b869ad1e6603f08199f951e84b1f8c79cd3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/ac061819j$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/ac061819j$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,778,782,2754,27065,27913,27914,56727,56777</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18737633$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17408242$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Arduini, Fabiana</creatorcontrib><creatorcontrib>Errico, Ilenia</creatorcontrib><creatorcontrib>Amine, Aziz</creatorcontrib><creatorcontrib>Micheli, Laura</creatorcontrib><creatorcontrib>Palleschi, Giuseppe</creatorcontrib><creatorcontrib>Moscone, Danila</creatorcontrib><title>Enzymatic Spectrophotometric Method for Aflatoxin B Detection Based on Acetylcholinesterase Inhibition</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>A new method for aflatoxin B (AFB) determination is proposed. The AFB determination is based on acetylcholinesterase (AChE) inhibition, and the AChE residual activity is determined using the colorimetric method (Ellman's method). Cholinesterases (ChEs) from various sources were tested using AFB1 as reference aflatoxin. AChE from electric eel has shown the highest sensitivity to AFB1, and it was chosen for the rest of the work. To select and optimize the analytical procedures, an investigation on the type of AChE inhibition by AFB1 was carried out. The AChE degree of inhibition by AFB1 was independent of the incubation time and the enzyme concentrations, showing the reversibility of the inhibition. This reversibility of the inhibition permits a rapid analysis of AFB1, requiring only 3 min. For the development of the AFB1 assay, the pH, the time of reaction, temperature, and substrate concentration were evaluated and optimized. The linear range of 10−60 ng mL-1 was determined. To evaluate the selectivity of this method, the cross-reactivity with other aflatoxins such as aflatoxin B2, aflatoxin G1, aflatoxin G2, and aflatoxin M1 was investigated. Finally, the suitability of the assay for AFB1 quantification in barley was evaluated. This study shows a new approach to detect aflatoxins based on enzyme inhibition and has advantages such as the ease of use, rapidity, and cost effectiveness. Thus, it could find a possible use as a screening method for this type of mycotoxins.</description><subject>Acetylcholinesterase - chemistry</subject><subject>Acetylcholinesterase - drug effects</subject><subject>Aflatoxin B1 - analysis</subject><subject>Aflatoxin B1 - pharmacology</subject><subject>Analytical chemistry</subject><subject>Animals</subject><subject>Chemistry</subject><subject>Electrophorus</subject><subject>Enzyme Activation - drug effects</subject><subject>Exact sciences and technology</subject><subject>Methanol - chemistry</subject><subject>Methanol - pharmacology</subject><subject>Molecular Structure</subject><subject>Sensitivity and Specificity</subject><subject>Spectrometric and optical methods</subject><subject>Spectrophotometry - methods</subject><subject>Structure-Activity Relationship</subject><subject>Temperature</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpt0EtLAzEQB_AgitbHwS8ge_HgYXWyj2z2WKtWoaLg8xZmswlN3W5KEsH66Y202IunDJkfw8yfkGMK5xQyeoESGOW0nm2RAS0zSBnn2TYZAECeZhXAHtn3fgZAKVC2S_ZoVQDPimxA9HX_vZxjMDJ5WigZnF1MbbBzFVz8uldhattEW5cMdYfBfpk-uUyuVIjU2FijV20Si6FUYdnJqe1Mr3xQLjaSu35qGvMLD8mOxs6ro_V7QF5urp9Ht-nkYXw3Gk5SLIoqpA3WjWwANY-bcy5Vg2WOwIocWygbzmpsqWIMcg3x3lrXJVW8aKjmsqplmx-Qs9Vc6az3TmmxcGaObikoiN-sxF9W0Z6s7OKzmat2I9fhRHC6BugldtphL43fOF7lFcvz6NKVM_Hwr78-ug_BIinF8-OTGBejyfsrvInxZi5KL2b20_Uxkn8W_AG2N44K</recordid><startdate>20070501</startdate><enddate>20070501</enddate><creator>Arduini, Fabiana</creator><creator>Errico, Ilenia</creator><creator>Amine, Aziz</creator><creator>Micheli, Laura</creator><creator>Palleschi, Giuseppe</creator><creator>Moscone, Danila</creator><general>American Chemical Society</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>20070501</creationdate><title>Enzymatic Spectrophotometric Method for Aflatoxin B Detection Based on Acetylcholinesterase Inhibition</title><author>Arduini, Fabiana ; Errico, Ilenia ; Amine, Aziz ; Micheli, Laura ; Palleschi, Giuseppe ; Moscone, Danila</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a447t-ba9bcb0af800388ceba53a0643ad05b869ad1e6603f08199f951e84b1f8c79cd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Acetylcholinesterase - chemistry</topic><topic>Acetylcholinesterase - drug effects</topic><topic>Aflatoxin B1 - analysis</topic><topic>Aflatoxin B1 - pharmacology</topic><topic>Analytical chemistry</topic><topic>Animals</topic><topic>Chemistry</topic><topic>Electrophorus</topic><topic>Enzyme Activation - drug effects</topic><topic>Exact sciences and technology</topic><topic>Methanol - chemistry</topic><topic>Methanol - pharmacology</topic><topic>Molecular Structure</topic><topic>Sensitivity and Specificity</topic><topic>Spectrometric and optical methods</topic><topic>Spectrophotometry - methods</topic><topic>Structure-Activity Relationship</topic><topic>Temperature</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Arduini, Fabiana</creatorcontrib><creatorcontrib>Errico, Ilenia</creatorcontrib><creatorcontrib>Amine, Aziz</creatorcontrib><creatorcontrib>Micheli, Laura</creatorcontrib><creatorcontrib>Palleschi, Giuseppe</creatorcontrib><creatorcontrib>Moscone, Danila</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Arduini, Fabiana</au><au>Errico, Ilenia</au><au>Amine, Aziz</au><au>Micheli, Laura</au><au>Palleschi, Giuseppe</au><au>Moscone, Danila</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Enzymatic Spectrophotometric Method for Aflatoxin B Detection Based on Acetylcholinesterase Inhibition</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2007-05-01</date><risdate>2007</risdate><volume>79</volume><issue>9</issue><spage>3409</spage><epage>3415</epage><pages>3409-3415</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>A new method for aflatoxin B (AFB) determination is proposed. The AFB determination is based on acetylcholinesterase (AChE) inhibition, and the AChE residual activity is determined using the colorimetric method (Ellman's method). Cholinesterases (ChEs) from various sources were tested using AFB1 as reference aflatoxin. AChE from electric eel has shown the highest sensitivity to AFB1, and it was chosen for the rest of the work. To select and optimize the analytical procedures, an investigation on the type of AChE inhibition by AFB1 was carried out. The AChE degree of inhibition by AFB1 was independent of the incubation time and the enzyme concentrations, showing the reversibility of the inhibition. This reversibility of the inhibition permits a rapid analysis of AFB1, requiring only 3 min. For the development of the AFB1 assay, the pH, the time of reaction, temperature, and substrate concentration were evaluated and optimized. The linear range of 10−60 ng mL-1 was determined. To evaluate the selectivity of this method, the cross-reactivity with other aflatoxins such as aflatoxin B2, aflatoxin G1, aflatoxin G2, and aflatoxin M1 was investigated. Finally, the suitability of the assay for AFB1 quantification in barley was evaluated. This study shows a new approach to detect aflatoxins based on enzyme inhibition and has advantages such as the ease of use, rapidity, and cost effectiveness. Thus, it could find a possible use as a screening method for this type of mycotoxins.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>17408242</pmid><doi>10.1021/ac061819j</doi><tpages>7</tpages></addata></record> |
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subjects | Acetylcholinesterase - chemistry Acetylcholinesterase - drug effects Aflatoxin B1 - analysis Aflatoxin B1 - pharmacology Analytical chemistry Animals Chemistry Electrophorus Enzyme Activation - drug effects Exact sciences and technology Methanol - chemistry Methanol - pharmacology Molecular Structure Sensitivity and Specificity Spectrometric and optical methods Spectrophotometry - methods Structure-Activity Relationship Temperature |
title | Enzymatic Spectrophotometric Method for Aflatoxin B Detection Based on Acetylcholinesterase Inhibition |
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