NMR Immunosensor Based on a Targeted Gadolinium Nanoprobe for Detecting Salmonella in Milk.

Detecting harmful pathogens in food is not only a crucial aspect of food quality management but also an effective way to ensure public health. In this paper, a complete nuclear magnetic resonance biosensor based on a novel gadolinium (Gd)-targeting molecular probe was developed for the detection of Salmonella in milk. First, streptavidin was conjugated to the activated macromolecular polyaspartic acid (PASP) via an amide reaction to generate SA-PASP. Subsequently, the strong chelating and adsorption properties of PASP toward the lanthanide metal gadolinium ions were exploited to generate the magnetic complex (SA-PASP-Gd). Finally, the magnetic complex was linked to biotinylated antibodies to obtain the bioprobe and achieve the capture of Salmonella. Under optimal experimental conditions, the sensor we have constructed can achieve a rapid detection of Salmonella within 1.5 h, with a detection limit of 7.1 × 103 cfu mL-1.

Rapid detection of Salmonella in milk by labeling-free electrochemical immunosensor based on an Fe3O4-ionic liquid-modified electrode.

Electrochemical sensors show distinct advantages over other types of sensors in the rapid detection of microorganisms. Here, we attempted to construct a label-free electrochemical immunosensor based on an Fe3O4-ionic liquid (IL)-modified electrode to rapidly detect Salmonella in milk. The excellent ionic conductivity of the IL facilitated sensor construction, and the large surface area of nano-Fe3O4 provided numerous sites for subsequent experiments. An antibody was fixed on the Fe3O4-IL complex with polyglutamic acid modification by a simple infusion method. The microstructure of the Fe3O4-IL composites was investigated by scanning electron microscopy, and the elements and structures of the composites were analyzed by energy dispersive X-ray and Fourier transform infrared spectroscopy. Under optimized experimental conditions, the detection range of the constructed sensor was 3.65 × 102-3.65 × 108 CFU mL-1, and the LOD was 1.12 × 102 CFU mL-1 (S/N = 3). In addition, the prepared electrochemical immunosensor is convenient for detecting foodborne pathogens because of its outstanding stability, good selectivity, and repeatability.

Study on the effect of different concentrations of choline glycine ionic liquid-water mixtures on debranched starch butyrylation reaction.

In this study, the effect of choline glycine ionic liquids on the butyrylation of starch was investigated by the butyrylation of debranched cornstarch in different concentrations of choline glycine ionic liquid-water mixtures (choline glycine ionic liquids to water in mass ratios of 0:10, 4:6, 5:5, 6:4, 7:3, 8:2 and 10:0). The butyryl characteristic peaks in 1H NMR and FTIR of the butyrylated samples indicated the success of butyrylation modification. 1H NMR calculations showed that the most effective mass ratio of choline glycine ionic liquids to water (6:4) increased the butyryl substitution degree from 0.13 to 0.42. X-ray diffraction results showed that the crystalline type of the starch modified in the choline glycine ionic liquid-water mixtures changed from B-type to a mixture of V-type and B-type isomers. The butyrylated starch modified in the ionic liquid increased its own content of resistant starch from 25.42 % to 46.09 %. This study highlights the effect of different concentrations of choline glycine ionic liquid-water mixtures on the promotion of starch butyrylation reactions.

A fluorescent biosensor based on quantum dot-labeled streptavidin and poly-l-lysine for the rapid detection of Salmonella in milk.

Salmonella, as a common foodborne pathogen in dairy products, poses a great threat to human health. We studied a new detection method based on quantum dots (QD). A fluorescent biosensor with multiple fluorescent signal amplification based on a streptavidin (SA) biotin system and the polyamino linear polymer poly-l-lysine (PLL) were established to detect Salmonella in milk. First, Salmonella was captured on a black 96-well plate with paired Salmonella mAb to form a double-antibody sandwich. Second, SA was immobilized on biotin-modified mAb by SA-biotin specific bond. Then, the biotin-modified polylysine (BT-PLL) was bound on SA and specifically bonded again through the SA-biotin system. Finally, water-soluble CdSe/ZnS QD-labeled SA was added to a black 96-well plate for covalent coupling with BT-PLL. The fluorescent signal was amplified in a dendritic manner by the layer-by-layer overlap of SA and biotin and the covalent coupling of biotinylated PLL. Under optimal conditions, the detection limit was 4.9 × 103 cfu/mL in PBS. The detection limit was 10 times better than that of the conventional sandwich ELISA. In addition, the proposed biosensor was well specific and could be used for detecting Salmonella in milk samples.

A novel electrochemical immunosensor based on Fe3O4@graphene nanocomposite modified glassy carbon electrode for rapid detection of Salmonella in milk.

Foods contaminated by foodborne pathogens have always been a great threat to human life. Herein, we constructed an electrochemical immunosensor for Salmonella detection by using a Fe3O4@graphene modified electrode. Because of the excellent electrical conductivity and mechanical stability of graphene and the large specific surface area of Fe3O4, the Fe3O4@graphene nanocomposite exhibits an excellent electrical signal, which greatly increased the sensitivity of the immunosensor. Gold nanoparticles were deposited on Fe3O4@graphene nanocomposite by electrochemical technology for the immobilization of the antibody. Cyclic voltammetry was selected to electrochemically characterize the construction process of immunosensors. The microstructure and morphology of related nanocomposites were analyzed by scanning electron microscopy. Under optimized experimental conditions, a good linear relationship was achieved in the Salmonella concentration range of 2.4 × 102 to 2.4 × 107 cfu/mL, and the limit of detection of the immunosensor was 2.4 × 102 cfu/mL. Additionally, the constructed immunosensor exhibited acceptable selectivity, reproducibility, and stability and provides a new reference for detecting pathogenic bacteria in milk.

The mechanism of delaying starch digestion by luteolin.

In this study, the mechanisms of the delay of starch digestion by luteolin were revealed by studying the luteolin-PPA (porcine pancreatic α-amylase) interaction and luteolin-starch interaction. The luteolin-PPA interaction was investigated by inhibitory kinetics analysis, fluorescence quenching, circular dichroism (CD), Fourier transform infrared (FT-IR) spectroscopy and molecular docking. The results of the inhibitory kinetics revealed that luteolin was a mixed-type inhibitor of PPA and that the inhibitory action was reversible. Fluorescence spectroscopy (including fluorescence quenching and thermodynamics) and molecular docking analyses indicated that hydrogen bonds and hydrophobic forces were the main forces between PPA and luteolin. CD and FT-IR spectroscopy analyses showed that the interaction between luteolin and PPA changed the secondary structure of PPA and induced a decline in its activity. In addition, the luteolin-starch interaction was also studied using UV-visible absorption and X-ray diffraction analyses. These indicated that luteolin could bind with PPA, and that hydrogen bonds and van der Waals forces may be present. Overall, luteolin delayed starch digestion not only by binding with PPA but also by binding with starch. Thus, luteolin has the potential to prevent and control diabetes by being added into starch-based food to delay starch digestion.

Rapid detection of Salmonella in milk by a nuclear magnetic resonance biosensor based on the streptavidin-biotin system and O-carboxymethyl chitosan target gadolinium probe.

Rapid and sensitive detection of foodborne pathogens is of great importance for food safety. Here, a set of nuclear magnetic resonance (NMR) biosensors based on a O-carboxymethyl chitosan target gadolinium (Gd) probe was developed to quickly detect Salmonella in milk by combining NMR technology and bioimmunotechnology with membrane filtration technology. First, O-carboxymethyl chitosan (O-CMC) was biotinylated to prepare biotinylated O-carboxymethyl chitosan (biotin-O-CMC) through amide reaction, and biotinylated magnetic complexes (biotin-O-CMC-Gd) were obtained by using O-CMC, which has strong chelating adsorption on Gd. The target probe was obtained by combining biotin-O-CMC-Gd with the biotinylated antibody (biotin-antibody) via streptavidin (SA) by introducing the SA-biotin system. Then, Salmonella was captured by the target probe through antigen-antibody interaction. Finally, NMR was used to measure the longitudinal relaxation time (T1) of the filtrate collected by membrane filtration. This NMR biosensor with good specificity and high efficiency can detect Salmonella with the sensitivity of 1.8 × 103 cfu/mL within 2 h; in addition, it can realize the detection of complex samples because of its strong anti-interference capability and may open up a new method for rapid detection of Salmonella, which has a great application potential.

Rapid detection of Salmonella in milk with a nuclear magnetic resonance biosensor based on a streptavidin-biotin system and a polyamidoamine-dendrimer-targeted gadolinium probe.

Rapid and sensitive detection technology is the key to preventing food-borne disease outbreaks. In this study, a low-field nuclear magnetic resonance (NMR) biosensor based on polyamidoamine dendrimers was prepared for the rapid detection of Salmonella in milk. The polyamidoamine dendrimer was biotinylated by amide reaction and chelated to diethylene triamine pentacetate acid and gadolinium to form magnetic complexes. The antibody and magnetic complexes were combined through a streptavidin-biotin system using streptavidin as an intermediate bridge to obtain the immunoprobe. Salmonella was captured by the immunoprobe via antigen-antibody interaction and then separated from the mixture by membrane filtration. Finally, the longitudinal relaxation signal of the filtrate was obtained by NMR. The biosensor had excellent anti-interference capability and could detect Salmonella within 1.5 h at a sensitivity of 103 cfu mL-1. This method based on NMR can realize detection in complex samples and has the potential to be a quick and nondestructive method for detecting target bacteria.

Effects of xanthan and konjac gums on pasting, rheology, microstructure, crystallinity and in vitro digestibility of mung bean resistant starch.

The effects of different concentrations of xanthan and konjac gums on the pasting, rheological properties, microstructure, crystallinity, and digestibility of mung bean resistant starch (MRS) were investigated. Based on the results of pasting properties, the adjunction of gums increased the peak, breakdown, and final viscosities of resistant starch. Compared with resistant starch, the addition of gum significantly increased the K value and dynamic moduli (G', G") of MRS with increasing gum concentration. This finding indicates that the mixtures had higher viscoelasticity. Mixtures with xanthan gum of MRS had larger starch particle compared with MRS, as revealed by SEM. All starches showed B and V-type crystallinity with high crystallinity. MRS had the highest summation of resistant starch (RS) and slowly digestible starch (SDS) of 71.89%. MRS had the lowest hydrolysis rate, which obviously decreased from 71.89% to 57.71% with increasing konjac gum from 0 to 0.30%.

Determination of chlorine ions in raw milk by pulsed amperometric detection in a flow injection system.

Chloride ion concentration in milk was determined by pulsed amperometric detection in a flow injection system. Results showed that the Au electrode lost 3 electrons at 1.10 V and formed chloroaurate ions (AuCl4-) by combining with chloride ions, after which AuCl4- was partly reduced to Au at 0.6 V. Based on the electrochemical process, a triple waveform with detection potential of 1.15 V, detection time of 150 ms, oxidation potential of 1.4 V, oxidation time of 550 ms, reduction potential of 0 V, and reduction time of 400 ms was applied to the Au electrode for detecting chloride ion concentration in milk. The approach is rapid and automatic and features a detection limit of 0.005 g/L. The relative standard deviation obtained by 60 repetitive injections reached 1.48% at 2 g/L of NaCl. The method developed using the Au electrode without modification was used to analyze the chloride ion concentration in raw milk without preprocessing. The method showed good agreement with potentiometric titration.