McNamee Sara E, Rosar Giulia, Persic Lidija, Elliott Christopher T, Campbell Katrina
a Institute for Global Food Security, School of Biological Sciences , Queen's University , Belfast , UK.
b Tecna s.r.l ., Trieste , Italy.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2017 Apr;34(4):562-572. doi: 10.1080/19440049.2017.1280188. Epub 2017 Feb 16.
Practical solutions for multiple antibiotic determination in food are required by the food industry and regulators for cost-effective screening purposes. This study describes the feasibility in development and preliminary performance of a novel multispot nanoarray for antibiotic screening in honey. Using a multiplex approach, the metabolites of the four main nitrofuran antibiotics, including morpholinomethyl-2-oxazolidone (AMOZ), 3-amino-2-oxazolidinone (AOZ), semicarbazide (SEM), 1-aminohydantoin (AHD) and chloramphenicol (CAP), were simultaneously detected. Antibodies specific to the five antibiotics were nano-spotted onto microtitre plate wells and a direct competitive assay format was employed. The assay characteristics and performance were evaluated for feasibility as a screening tool for antibiotic determination in honey to replace traditional ELISAs. Optimisation of the spotting and assay parameters was undertaken with both individual and multiplex calibration curves generated in PBS and a honey matrix. The limits of detection as determined by the 20% inhibitory concentrations (IC) were determined as 0.19, 0.83, 0.09, 15.2 and 35.9 ng ml in PBS, 0.34, 0.87, 0.17, 42.1 and 90.7 ng ml in honey (fortified at the start of the extraction), and 0.23, 0.98, 0.24, 24.8 and 58.9 ng ml in honey (fortified at the end of the extraction) for AMOZ, AOZ, CAP, SEM and AHD respectively. This work has demonstrated the potential of multiplex analysis for antibiotics with results available for 40 samples within a 90-min period for antibiotics sharing a common sample preparation. Although both the SEM and AHD assay do not show the required sensitivity with the antibodies available for use to meet regulatory limits, with further improvements in these particular antibodies this multiplex format has the potential to show a reduction in cost with reduced labour time in combination with the high-throughput screening of samples. This is the first 96-well spotted microtitre plate nanoarray for the semi-quantitative and simultaneous analysis of antibiotics.
食品行业和监管机构出于经济高效的筛选目的，需要用于食品中多种抗生素测定的实用解决方案。本研究描述了一种用于蜂蜜中抗生素筛选的新型多点纳米阵列在开发方面的可行性及其初步性能。采用多重分析方法，同时检测了四种主要硝基呋喃抗生素的代谢物，包括吗啉甲基 - 2 - 恶唑烷酮（AMOZ）、3 - 氨基 - 2 - 恶唑烷酮（AOZ）、氨基脲（SEM）、1 - 氨基乙内酰脲（AHD）和氯霉素（CAP）。将针对这五种抗生素的特异性抗体纳米点样到微孔板孔中，并采用直接竞争检测形式。评估了该检测方法的特性和性能，以确定其作为蜂蜜中抗生素测定筛选工具替代传统酶联免疫吸附测定（ELISA）的可行性。利用在磷酸盐缓冲盐水（PBS）和蜂蜜基质中生成的单独和多重校准曲线，对斑点和检测参数进行了优化。由20%抑制浓度（IC）确定的检测限在PBS中分别为：AMOZ为0.19 ng/ml、AOZ为0.83 ng/ml、CAP为0.09 ng/ml、SEM为15.2 ng/ml、AHD为35.9 ng/ml；在蜂蜜（提取开始时添加）中分别为：AMOZ为0.34 ng/ml、AOZ为0.87 ng/ml、CAP为0.17 ng/ml、SEM为42.1 ng/ml、AHD为90.7 ng/ml；在蜂蜜（提取结束时添加）中分别为：AMOZ为0.23 ng/ml、AOZ为
