Simultaneous detection of foodborne pathogenic bacteria in milk by fluorescence immunoassay.

Rapid, sensitive and simultaneous detection of multiple bacteria in foodborne is still a major challenge in public health field. Here, a fluorescence immunoassay that can achieve high-throughput detection of three Gram-positive foodborne pathogenic bacteria simultaneously was proposed. Vancomycin and bovine serum albumin conjugate (Van-BSA) was immobilized on a polycarbonate chip to capture three Gram-positive foodborne pathogenic bacteria, Staphylococcus aureus (S. aureus), Bacillus cereus (B. cereus) and Listeria monocytogenes (L. monocytogenes). CdSe/ZnS quantum dot modified antibodies (Ab-QD) were prepared by carbodiimide coupling chemistry. Due to the affinity reaction between antibodies and proteins on the bacterial surface, the simultaneous detection of multiple Gram-positive bacteria was achieved by monitoring the fluorescence signal of quantum dot by a portable microfluidic chip analyzer. Under optimal conditions, low detection limits was 18 CFU/well, 3 CFU/well and 36 CFU/well for S. aureus, B. cereus and L. monocytogenes, respectively. With satisfactory accuracy and precision, the proposed fluorescence immunoassay holds good prospects to detect pathogens in real food samples.

Fluorescence sensor for organophosphorus pesticide detection based on the alkaline phosphatase-triggered reaction.

The threat of organophosphorus pesticide (OPP) residue to food safety and human health has caused widespread concern. In this paper, a sensitive fluorescence sensor for OPP detection was constructed based on the alkaline phosphatase (ALP) -triggered in situ reaction. In this method, ALP catalyses the dephosphorylation of the substrate l-ascorbic acid 2-phosphate sesquimagnesium salt hydrate (AAP) to generate l-ascorbic acid (AA). AA instantly combines with o-phenylenediamine (OPD) to form 3-(1,2-dihydroxyethyl)furo[3,4-b]quinoxalin-1(3H)-one (DFQ), which contains a quinoxaline core skeleton fluorophore and emits a strong fluorescence intensity at 425 nm. The existence of OPPs inhibits the activity of ALP and the production of AA and DFQ. As a result, the fluorescence intensity obviously decreases. Under optimal conditions, the fluorescence intensity linearly depends on the logarithm of chlorpyrifos concentration over a wide range of 20 pg/mL ∼1000 ng/mL with a detection limit of 15.03 pg/mL (S/N = 3), which is lower than the previously reported values. The sensor with its satisfactory accuracy and precision has been successfully applied to the detection of chlorpyrifos in leeks and celery samples with recoveries of 94.5-106.7% and an inter-assay relative standard deviation (RSD) below 11.51%. OPPs can be semiquantitatively determined by the colour changes in ultraviolet light. The superiority of the sensor is due to its visual simplicity without complex fluorescence labelling procedures and costly instruments.

Photoelectrochemical TiO2 nanotube arrays biosensor for asulam determination based on in-situ generation of quantum dots.

Inspired by the photoelectrochemical (PEC) properties of TiO2 nanotubes arrays (TNA) and their application as a super vessel for immobilizing biomolecules, we constructed an inhibition-effect PEC biosensor for determination of asulam based on the in-situ generation of CdS quantum dots (QDs) on TNA using an enzymatic reaction. Horseradish peroxidase (HRP) enzyme was covalently assembled on the inner-wall of TNAs, which exhibited good electrochemical and catalytic properties. In the mixture solution containing H2O2, CdY and S2O32-, HRP enzyme in TNAs catalyzed H2O2 reduce S2O32- to S2-. The generated S2- reacted with CdY to form CdS QDs in situ on the TNAs, improving the PEC performance of TNA under visible light irradiation. The photocurrent would decrease after addition of asulam due to its inhibitory effect towards HRP enzyme activity. Under the optimal experimental conditions, the constructed PEC TNA/HRP biosensor exhibited a satisfying linear range (0.02-2.0 ng mL-1), low limit of detection (4.1 pg mL-1) and good selectivity towards asulam determination, and has been successfully applied for the analysis of real environmental water samples with good accuracy of the recoveries ranged from 90% to 114%.

N-Heterocyclic Carbene-Catalyzed [4 + 2] Cyclization of Saturated Carboxylic Acid with o-Quinone Methides through in Situ Activation: Enantioselective Synthesis of Dihydrocoumarins.

An N-heterocyclic carbene (NHC)-catalyzed formal [4 + 2] synthesis of dihydrocoumarins was realized from saturated carboxylic acids and o-quinone methides via an in situ activation strategy. This protocol results in excellent diastereoselectivity and enantioselectivity and good yields and uses readily available and inexpensive starting materials.