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A Suitable Immunosensor for Chloramphenicol Determination: Study of Two Different Competitive Formats

[ Vol. 16 , Issue. 4 ]

Author(s):

Elisabetta Martini, Mauro Tomassetti*, Riccardo Angeloni, Mauro Castrucci and Luigi Campanella   Pages 412 - 423 ( 12 )

Abstract:


Background: A deep analytical study was performed on two different formats based on a “competitive” ELISA-type assay to develop a suitable, sensitive and cheap immune device for chloramphenicol determination that could be advantageously applied to the analysis of real matrices (pharmaceutical, food and environmental).

Methods: To this purpose peroxidase enzyme as a marker and an amperometric electrode for hydrogen peroxide, as a transducer, were used. Through the first competitive format, chloramphenicol determination was based on the competition between chloramphenicol and conjugated with biotin-avidinperoxidase chloramphenicol, both free in solution, for anti-chloramphenicol immobilized in the membrane, while the second competitive format was based on the competition between free in solution chloramphenicol and immobilized in membrane one, for anti-chloramphenicol biotin-avidin-peroxidase conjugated free in solution.

Results: The immunosensor was optimized by comparing the two used different “competitive” working formats on the basis of respective Kaff values, that were found to be about 105 and 104 (mol L-1)-1. The developed immune device displayed good selectivity for Chloramphenicol and LOD (limit of detection) was of the order of 10-9 mol L-1. The immunosensor was also used to test the presence of Chloramphenicol in real matrices such as cow milk, river wastewater and pharmaceutical formulations; recovery tests, using the standard addition method, gave satisfactory results.

Conclusion: The results proved the validity of this immune device based on the competition between chloramphenicol and conjugated chloramphenicol obtained using biotin-avidin-peroxidase format, by which it is possible to carry out the analysis of chloramphenicol in milk and in river waste-waters with a % RSD ≤ 5 and with recovery values between 96% and 103%.

Keywords:

Chloramphenicol antibiotic, immunosensor, competitive format method optimization, cow milk, river waste-water, pharmaceutical formulation, analysis.

Affiliation:

Department of Chemistry, University of Rome “Sapienza”, P.le A. Moro 5, Rome 00185, Department of Chemistry, University of Rome “Sapienza”, P.le A. Moro 5, Rome 00185, Department of Chemistry, University of Rome “Sapienza”, P.le A. Moro 5, Rome 00185, Department of Chemistry, University of Rome “Sapienza”, P.le A. Moro 5, Rome 00185, Department of Chemistry, University of Rome “Sapienza”, P.le A. Moro 5, Rome 00185

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