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We proposed the first application of an electrophoretically mediated microanalysis (EMMA) method for fast online discrimination and determination of substrate enantiomers, which was achieved by just one EMMA assay. Lactate dehydrogenase (LDH)-catalyzed reaction was studied to evaluate the feasibility and performance of the presented method. The L- and D-LDH chiral enzymatic reactions, which are highly stereoselective to the lactate enantiomers, were initiated successively in one capillary, and the corresponding products, nicotinamide adenine dinucleotide (NADH), were online discriminated and detected by UV absorption. Excellent linear dependence of the two NADH peak intensities on the concentration of the corresponding lactate enantiomers was obtained within a wide range of 0.1-10 mM. The limit of detection was 26 μM for D-lactate and 49 μM for L-lactate (S/N = 3). Good repeatability of online chiral discrimination was demonstrated with relative standard deviation (RSD) < 6.3% for NADH peak height and RSD < 1.5% for migration time (n = 5). K(m) values for L- and D-lactate were measured and were consistent with those of the off-line enzyme assays. The presented method was successfully applied to determine the L-/D-lactate in several yogurt and wine samples. Our study shows a new application of the EMMA method utilizing high stereoselectivity of enzymes for fast online chiral analysis.

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We describe herein a newly developed chemiluminescent optical fiber immunosensor (OFIS) with a tunable detection range for multiplexed analysis of veterinary drug residues with vastly different concentrations in milk samples. The optical fiber probe is used as a carrier of biorecognition element as well as a transducer, enabling a low-cost compact design, which makes this system suitable for cost-effective on-site detection of the target analytes. Importantly, the synergy between modulation of the length of the optical fiber sensing region and the number of fibers allows performing multiplexed immunoassays in an easily controllable manner over a tunable detection range from pg/mL to μg/mL analyte concentrations. By combining the optical fiber sensor with a nanocomplex signal amplification system, a highly sensitive chemiluminescent OFIS system is demonstrated for the multiplexed assaying of veterinary drug residues in milk samples with linear ranges of 10–(2 × 104) pg/mL for chloramphenicol, 0.5–500 ng/mL for sulfadiazine, and 0.1–300 μg/mL for neomycin. This controllable strategy, based on modulation of the fiber probe, provides a versatile platform for multiplexed quantitative detection of both low-abundance and high-abundance targets, which shows great potential for on-site testing in food safety.