Simultaneous detection of four nitrofuran metabolites in honey using high-throughput suspension array technology

Abstract

In this study, a new suspension array technology is proposed for the simultaneous quantitative detection of four major metabolites [3-amino-5-methylmorpholino-2-oxazolidinone (AMOZ), 3-amino-2oxazolidinone (AOZ), semicarbazide (SEM), and 1-aminohydantoin (AHD)] of nitrofurans in honey samples. The analytes were coupled to bovine serum albumin before coupling to the microspheres. After coupling, a mixture of biotinylated antibodies specific for the four analytes, along with the analyte standard solutions or samples, was added to microtiter tubes. Then, the microtiter tubes were incubated at 37 C for 45 min before streptavidin-R-phycoerythrin was added. Analyte residues were quantitatively analyzed by detecting the mean fluorescence intensity. The detection ranges for the four analytes were 0.02-40 mg kg À1 and the detection limit of AMOZ, AOZ, SEM, and AHD in honey samples were 0.087, 0.031, 0.055, and 0.131 mg kg À1 , respectively. The recovery rates ranged from 76.6% to 106.1% for fortified samples at levels of 0.25, 1.0, 5.0, and 20 mg kg À1 with coefficient of variation values <15%. Finally, this method was compared to commercially available kits using fortified and 'blind' honey samples and the measurements obtained using these two methods correlated well. These results indicate that the suspension array method is suitable for detecting AMOZ, AOZ, SEM, and AHD, as well as other potential drug residues in animal tissues.

The Influence of Chemical Contaminants on the Physicochemical Properties of Unifloral and Multifloral Honey

“…Chloramphenicol and nitrofuran residues in honey can be determined by the ELISA technique, and nitroimidazole residues by liquid chromatography coupled with mass spectrometry [57][58][59]. MRLs for nitrofurans and their metabolites analyzed from honey samples are established by the European Union through regulation.…”

Development of Antibodies and Immunoassays for Monitoring of Nitrofuran Antibiotics in the Food Chain

“…Luminex Xmap Suspension Array technology has been used for the simultaneous quantification of AMOZ, AOZ, SEM and AHD in honey [63]. The analytes were coupled to BSA before coupling to carboxylated polystyrene microspheres.…”

“…Muscle is generally the target of choice for monitoring nitrofuran residues in the food chain. Immunoassays are also described for liver [7,19,45,54], honey [18,23,40,42,55,58,62,63], egg [18,34,42,59], intestine/casings [18,23,40,55], milk powder [12], baby food [15], kidney [45], eyes [30], urine [19,25], plasma [54], water [29] and animal feed [27,28,31,32,56]. Table 2 that, if commercial supply and demand is a reliable indicator, microwell plate ELISA still remains the nitrofuran screening format of choice for those without access to sophisticated mass spectrometry facilities.…”

Cytometric Microbead Magnetic Suspension Array for High-Throughput Ultrasensitive Detection of Aflatoxin B1

“…This technology combines advanced fluidics, optics, and digital signal processing with microbead sets to detect multiple targets and is usually created by capturing analytes on the surface of microbeads encoded with distinct optical probes in conjunction with flow cytometric analysis. The large surface area of suspension arrays and the concentration of reactants on the surface of the microbead improve binding kinetics, reducing assay time and increasing assay sensitivity to provide economical and high-throughput analysis of a large number of samples or multiple analytes. − Recently, the introduction of fluorescence-encoded magnetic microbeads as the carriers has attracted increasing and wide interest because of their unique features, such as ease of synthesis and functionalization, unique fluorescent characteristics, good biocompatibility, and large surface area that facilitates covalent conjugation of various capture molecules including protein antigens, nucleic acids, and aptamers. , They also allow easy differentiation by size and fluorescence intensity, as well as rapid, controllable, and highly effective separation of multiple targets and high-throughput detection only needing samples in microliter or nanoliter quantity. , In addition, magnetic separation instead of complex centrifugation, purification, and enrichment steps can lower or avoid unpredictable loss of targets to improve the detection sensitivity and accuracy . As a result, magnetic suspension array has become a highly attractive platform for high-throughput analysis in drug screening, food and environmental monitoring, and other fields. , …”