Amphenicol residues in foods of animal origin can pose severe risks to human health;however,the determination of amphenicols is usually based on a single-mode immunoassays.So a multi-mode method capable of determining...Amphenicol residues in foods of animal origin can pose severe risks to human health;however,the determination of amphenicols is usually based on a single-mode immunoassays.So a multi-mode method capable of determining amphenicols is desirable.In this study,a tri-mode fluorescence detection method for amphenicols was developed on the basis of the ribosomal protein L16(rpL16)of Thermus thermophilus.In particular,rpL16 was expressed,and its recognition mechanism for three amphenicols was investigated using a molecular docking technique.The protein was conjugated to red quantum dots to prepare a recognition element,which was combined with a blue carbon dotlabeled fluorescent tracer to develop a direct competitive microplate-based assay for the detection of amphenicols.As the analyte concentration increased,there was a color change from purple to red,the RGB value increased,and the fluorescence intensity from the tracer decreased.The results could be read with the naked eye,a smartphone,and a multimode microplate reader.The limits of detection for determination of the three amphenicols in eggs were 1.0 ng/g via visual detection,10 pg/g via smartphone detection,and 0.05-0.17 pg/g via instrumental detection.This study reports the first tri-mode detection method for the determination of small molecule compounds based on quantum dots and carbon dots,providing a versatile tool for amphenicol detection in foods of animal origin under different usage scenarios.展开更多
Objective:Amphenicols(chloramphenicol,thiamphenicol and forfenicol)can cause aplastic anaemia and other severe side effects to consumers;therefore,it is necessary to inspect their residues in foods of animal origin.Ho...Objective:Amphenicols(chloramphenicol,thiamphenicol and forfenicol)can cause aplastic anaemia and other severe side effects to consumers;therefore,it is necessary to inspect their residues in foods of animal origin.However,there has been no report on the use of amphenicols receptor for the determination of their residues,and none of the previously reported immunoassays for amphenicols can differentiate the specifc species.Materials and Methods:In this study,the ribosomal protein L16 of Escherichia coli was frst expressed,and its intermolecular interaction mechanisms with the three amphenicols was studied using the molecular docking technique.The protein was then combined with three enzymelabelled conjugates to develop a direct competitive array on microplate for determination of the three drugs in egg.Results:Due to the use of principal component analysis to analyse the data,this method could discriminate the three drugs in the range 0.1–10 ng/mL,and the limits of detection for the three drugs were in the range of 0.0002–0.0009 ng/mL.The analysis results for the unknown egg samples were consistent with a liquid chromatography–tandem mass spectrometry method,and the method performances were superior to the previous immunoassays for amphenicols.Conclusion:This is the frst paper reporting the use of ribosomal protein L16 to develop a competitive array for discriminative determination of amphenicols in food samples.展开更多
基金support of Science and Technology Program of Hebei Administration for Market Regulation(No.2024ZC01).
文摘Amphenicol residues in foods of animal origin can pose severe risks to human health;however,the determination of amphenicols is usually based on a single-mode immunoassays.So a multi-mode method capable of determining amphenicols is desirable.In this study,a tri-mode fluorescence detection method for amphenicols was developed on the basis of the ribosomal protein L16(rpL16)of Thermus thermophilus.In particular,rpL16 was expressed,and its recognition mechanism for three amphenicols was investigated using a molecular docking technique.The protein was conjugated to red quantum dots to prepare a recognition element,which was combined with a blue carbon dotlabeled fluorescent tracer to develop a direct competitive microplate-based assay for the detection of amphenicols.As the analyte concentration increased,there was a color change from purple to red,the RGB value increased,and the fluorescence intensity from the tracer decreased.The results could be read with the naked eye,a smartphone,and a multimode microplate reader.The limits of detection for determination of the three amphenicols in eggs were 1.0 ng/g via visual detection,10 pg/g via smartphone detection,and 0.05-0.17 pg/g via instrumental detection.This study reports the first tri-mode detection method for the determination of small molecule compounds based on quantum dots and carbon dots,providing a versatile tool for amphenicol detection in foods of animal origin under different usage scenarios.
基金the National Natural Science Foundation of China(No.32372447)the Natural Science Foundation of Hebei Province(No.C2023204045),China.
文摘Objective:Amphenicols(chloramphenicol,thiamphenicol and forfenicol)can cause aplastic anaemia and other severe side effects to consumers;therefore,it is necessary to inspect their residues in foods of animal origin.However,there has been no report on the use of amphenicols receptor for the determination of their residues,and none of the previously reported immunoassays for amphenicols can differentiate the specifc species.Materials and Methods:In this study,the ribosomal protein L16 of Escherichia coli was frst expressed,and its intermolecular interaction mechanisms with the three amphenicols was studied using the molecular docking technique.The protein was then combined with three enzymelabelled conjugates to develop a direct competitive array on microplate for determination of the three drugs in egg.Results:Due to the use of principal component analysis to analyse the data,this method could discriminate the three drugs in the range 0.1–10 ng/mL,and the limits of detection for the three drugs were in the range of 0.0002–0.0009 ng/mL.The analysis results for the unknown egg samples were consistent with a liquid chromatography–tandem mass spectrometry method,and the method performances were superior to the previous immunoassays for amphenicols.Conclusion:This is the frst paper reporting the use of ribosomal protein L16 to develop a competitive array for discriminative determination of amphenicols in food samples.
