Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform
Ochratoxin A (OTA) is one of the most toxic naturally encountered contaminants and is found in a variety of foods and beverages, including cereals and wine. Driven by the strict regulations regarding the maximum allowable OTA concentration in foodstuff and the necessity for on-site determination, th...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-10-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/12/10/877 |
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| author | Chrysoula-Evangelia Karachaliou Georgios Koukouvinos Grigoris Zisis Dimosthenis Kizis Evangelia Krystalli George Siragakis Dimitris Goustouridis Sotirios Kakabakos Panagiota Petrou Evangelia Livaniou Ioannis Raptis |
| author_facet | Chrysoula-Evangelia Karachaliou Georgios Koukouvinos Grigoris Zisis Dimosthenis Kizis Evangelia Krystalli George Siragakis Dimitris Goustouridis Sotirios Kakabakos Panagiota Petrou Evangelia Livaniou Ioannis Raptis |
| author_sort | Chrysoula-Evangelia Karachaliou |
| collection | DOAJ |
| description | Ochratoxin A (OTA) is one of the most toxic naturally encountered contaminants and is found in a variety of foods and beverages, including cereals and wine. Driven by the strict regulations regarding the maximum allowable OTA concentration in foodstuff and the necessity for on-site determination, the development of fast and sensitive methods for the OTA determination in cereal flours and wine samples, based on white light reflectance spectroscopy, is presented. The method relied on appropriately engineered silicon chips, on top of which an OTA-protein conjugate was immobilized. A polyclonal antibody against OTA was then employed to detect the analyte in the framework of a competitive immunoassay; followed by the subsequent addition of a biotinylated secondary antibody and streptavidin for signal enhancement. A small size instrument performed all assay steps automatically and the bioreactions were monitored in real time as the software converted the spectral shifts into effective biomolecular adlayer thickness increase. The assay developed had a detection limit of 0.03 ng/mL and a working range up to 200 ng/mL. The assay lasted 25 min (less than 1h, including calibrators/antibody pre-incubation) and was accomplished following a simple sample preparation protocol. The method was applied to corn and wheat flour samples and white and red wines with recovery values ranging from 87.2 to 111%. The simplicity of the overall assay protocol and convenient instrumentation demonstrates the potential of the immunosensor developed for OTA detection at the point of need. |
| first_indexed | 2024-03-09T20:35:49Z |
| format | Article |
| id | doaj.art-427d0a2d94a6433ea68b5ee619b70487 |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-09T20:35:49Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-427d0a2d94a6433ea68b5ee619b704872023-11-23T23:12:06ZengMDPI AGBiosensors2079-63742022-10-01121087710.3390/bios12100877Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing PlatformChrysoula-Evangelia Karachaliou0Georgios Koukouvinos1Grigoris Zisis2Dimosthenis Kizis3Evangelia Krystalli4George Siragakis5Dimitris Goustouridis6Sotirios Kakabakos7Panagiota Petrou8Evangelia Livaniou9Ioannis Raptis10Immunopeptide Chemistry Lab., Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, National Centre for Scientific Research “Demokritos”, P.O. Box 60037, 15310 Agia Paraskevi, GreeceImmunoassay/Immunosensors Lab., Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, National Centre for Scientific Research “Demokritos”, P.O. Box 60037, 15310 Agia Paraskevi, GreeceInstitute of Nanoscience and Nanotechnology, National Centre for Scientific Research “Demokritos”, P.O. Box 60037, 15310 Agia Paraskevi, GreeceScientific Directorate of Phytopathology, Benaki Phytopathological Institute, 14561 Kifissia, GreeceYiotis Anonimos Emporiki & Viomixaniki Etaireia, 12131 Athens, GreeceTuv Austria Food Allergens Labs Ltd., Kalopsidas 38, 7060 Livadia, CyprusThetaMetrisis S.A., Christou Lada 40, 12132 Athens, GreeceImmunoassay/Immunosensors Lab., Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, National Centre for Scientific Research “Demokritos”, P.O. Box 60037, 15310 Agia Paraskevi, GreeceImmunoassay/Immunosensors Lab., Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, National Centre for Scientific Research “Demokritos”, P.O. Box 60037, 15310 Agia Paraskevi, GreeceImmunopeptide Chemistry Lab., Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, National Centre for Scientific Research “Demokritos”, P.O. Box 60037, 15310 Agia Paraskevi, GreeceInstitute of Nanoscience and Nanotechnology, National Centre for Scientific Research “Demokritos”, P.O. Box 60037, 15310 Agia Paraskevi, GreeceOchratoxin A (OTA) is one of the most toxic naturally encountered contaminants and is found in a variety of foods and beverages, including cereals and wine. Driven by the strict regulations regarding the maximum allowable OTA concentration in foodstuff and the necessity for on-site determination, the development of fast and sensitive methods for the OTA determination in cereal flours and wine samples, based on white light reflectance spectroscopy, is presented. The method relied on appropriately engineered silicon chips, on top of which an OTA-protein conjugate was immobilized. A polyclonal antibody against OTA was then employed to detect the analyte in the framework of a competitive immunoassay; followed by the subsequent addition of a biotinylated secondary antibody and streptavidin for signal enhancement. A small size instrument performed all assay steps automatically and the bioreactions were monitored in real time as the software converted the spectral shifts into effective biomolecular adlayer thickness increase. The assay developed had a detection limit of 0.03 ng/mL and a working range up to 200 ng/mL. The assay lasted 25 min (less than 1h, including calibrators/antibody pre-incubation) and was accomplished following a simple sample preparation protocol. The method was applied to corn and wheat flour samples and white and red wines with recovery values ranging from 87.2 to 111%. The simplicity of the overall assay protocol and convenient instrumentation demonstrates the potential of the immunosensor developed for OTA detection at the point of need.https://www.mdpi.com/2079-6374/12/10/877Ochratoxin Awhite light reflectance spectroscopy biosensorimmunoanalysiscereal flourswine |
| spellingShingle | Chrysoula-Evangelia Karachaliou Georgios Koukouvinos Grigoris Zisis Dimosthenis Kizis Evangelia Krystalli George Siragakis Dimitris Goustouridis Sotirios Kakabakos Panagiota Petrou Evangelia Livaniou Ioannis Raptis Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform Biosensors Ochratoxin A white light reflectance spectroscopy biosensor immunoanalysis cereal flours wine |
| title | Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform |
| title_full | Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform |
| title_fullStr | Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform |
| title_full_unstemmed | Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform |
| title_short | Fast and Accurate Determination of Minute Ochratoxin A Levels in Cereal Flours and Wine with the Label-Free White Light Reflectance Spectroscopy Biosensing Platform |
| title_sort | fast and accurate determination of minute ochratoxin a levels in cereal flours and wine with the label free white light reflectance spectroscopy biosensing platform |
| topic | Ochratoxin A white light reflectance spectroscopy biosensor immunoanalysis cereal flours wine |
| url | https://www.mdpi.com/2079-6374/12/10/877 |
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