RESUMEN
Glutathione S-transferase (GST) is a group of multifunctional enzyme and participates in many physiological processes, such as xenobiotic biotransformation, drug metabolism, and degradation of toxic products. Herein, we demonstrate a label-free fluorescent conjugated polymer nanoparticle (FCPNPs)-based single-particle enumeration (SPE) method for the sensitive GST assay. Fluorescence resonance energy transfer (FRET) is formed between the glutathione-modified FCPNPs (FCPNPs-GSH) and polyethylenimine-capped gold nanoparticles (GNPs@PEI). Therefore, the fluorescence of FCPNPs-GSH is quenched remarkably. In the presence of GST, GNPs@PEI stay away from FCPNPs-GSH due to the specific interaction between FCPNPs-GSH and GST, leading to the inhibition of FRET. As a result, the fluorescence emission of FCPNPs-GSH is restored, which is reflected as the increase of the number of fluorescent particles in the microscopic image. By statistically counting the target concentration-dependent fluorescent particle number, accurate quantification of GST is achieved. The linear range from 0.01 to 6 µg/mL is obtained for GST assay and the limit-of-detection (LOD) is 1.03 ng/mL, which is much lower than the ensemble fluorescence spectra measurements in bulk solution. In urine sample assay, satisfactory recoveries in the range of 97.5-106.5.0% are achieved. Because of the high sensitivity and excellent specificity, this method can be extended to the detection of other disease-related biomolecules in the future.
Asunto(s)
Transferencia Resonante de Energía de Fluorescencia , Colorantes Fluorescentes/química , Glutatión Transferasa/análisis , Nanopartículas/química , Polímeros/química , Glutatión Transferasa/metabolismoRESUMEN
Contamination of foods and feeds by aflatoxins is a universal yet serious problem all over the world. Particularly, aflatoxin B1 (AFB1) is the most primary form and readily leads to terrible damages to human health. In this work, we construct a sensitive aptasensor based on single-particle detection (SPD) to analyze AFB1 in peanut samples with luminescence resonance energy transfer (LRET) between the aptamer-modified upconversion nanoparticles (UCNPs-aptamer) and gold nanoparticles (GNPs). The UCNP-aptamer plays as the luminescence donor, while GNP acts as the energy acceptor. In the absence of AFB1, GNPs would adsorb onto the surface of UCNPs-aptamer because of the association between aptamers and GNPs, leading to luminescence quenching. However, the luminescence of UCNPs-aptamer is recovered gradually in the presence of AFB1, because the aptamers possess stronger affinity toward AFB1 than GNPs. Through statistically counting the number of luminescent particles on the glass slide surface, the concentration of AFB1 in solution is accurately determined. The linear dynamic range for AFB1 detection is from 3.13 to 125.00 ng/mL. The limit-of-detection (LOD) is 0.17 ng/mL, which is much lower than the allowable concentration in foods. As a result, this method would provide promising application for the sensitive detection of AFB1 in foods and feeds, which might make a meaningful contribution to food safety and public health in the future.
Asunto(s)
Aflatoxina B1/análisis , Aptámeros de Nucleótidos/química , Análisis de los Alimentos/métodos , Contaminación de Alimentos/análisis , Arachis/química , Transferencia Resonante de Energía de Fluorescencia/métodos , Análisis de los Alimentos/instrumentación , Oro/química , Límite de Detección , Luminiscencia , Nanopartículas del Metal/químicaRESUMEN
In a clinical assay, enzymes are essential biomarkers for human disease diagnosis. In this work, a spectral-resolved single-particle detection (SPD) method is introduced to quantify alkaline phosphatase (ALP) activity in human serum with a supraparticle (SP) based on MnO2-modified gold nanoparticle (denoted as GNP@MnO2 SP) as the probe. In the presence of ALP, 2-phospho-l-ascorbic acid trisodium salt can be hydrolyzed into l-ascorbic acid, which serves as a good reduction agent to trigger the decomposition of the MnO2 shell on the GNP surface. Given that a trace amount of ALP exists, noticeable scattering color change can be detected at the single-particle level due to the sensitive localized surface plasmon resonance (LSPR) effect from GNPs. With spectral-resolved dark-field optical microscopy, a linear dynamic range of 0.06 to 2.48 mU/mL ( R2 = 0.99) and a very low limit of detection of 5.8 µU/mL for the ALP assay are readily achieved, which is more sensitive over the methods based on ensemble sample measurement. As a consequence, this strategy opens a new avenue for the design of an ultrasensitive detection method for disease-correlated biomarker diagnosis in the future.
Asunto(s)
Fosfatasa Alcalina/sangre , Fosfatasa Alcalina/metabolismo , Resonancia por Plasmón de Superficie , Oro/química , Humanos , Compuestos de Manganeso/química , Nanopartículas del Metal/química , Óxidos/química , Tamaño de la Partícula , Propiedades de SuperficieRESUMEN
This study investigated the contents of saponins and phenolic compounds in relation to their antioxidant activity and α-glucosidase inhibition activity of 7 colored quinoa varieties. The total saponin content was significantly different among 7 varieties and ranged from 7.51 to 12.12 mg OAE/g DW. Darker quinoa had a higher content of phenolic compounds, as well as higher flavonoids and antioxidant activity than that of light varieties. Nine individual phenolic compounds were detected in free and bound form, with gallic acid and ferulic acid representing the major compounds. The free and bound phenolic compounds (gallic acid and ferulic acid in particular) exhibited high linear correlation with their corresponding antioxidant values. In addition, the free phenolic extracts from colored quinoa exhibited higher inhibitory activity against α-glucosidase than the bound phenolic extracts. These findings imply that colored quinoa with abundant bioactive phytochemicals could be an important natural source for preparing functional food.
Asunto(s)
Antioxidantes/farmacología , Chenopodium quinoa/química , Inhibidores de Glicósido Hidrolasas/farmacología , Fenoles/farmacología , Saponinas/farmacología , alfa-Glucosidasas/metabolismo , Animales , Antioxidantes/análisis , Flavonoides/análisis , Flavonoides/farmacología , Inhibidores de Glicósido Hidrolasas/análisis , Fenoles/análisis , Ratas , Saponinas/análisisRESUMEN
In this work, we demonstrate a convenient yet sensitive color-coded single-particle detection method for the quantification of pyrophosphate (PPi) by using single gold nanoparticle (GNP) as the probe. The design is based on GNP-dependent catalytic deposition of Cu onto the surface of GNPs with reduced nicotinamide adenine dinucleotide (NADH). Without PPi, Cu2+ can be directly reduced to Cu0 through the gold-catalyzed oxidization of NADH. In the presence of PPi, the coating process is impeded due to the strong coordination capability of PPi with Cu2+. The selective coating of Cu shell onto the GNPs surface results in the extraordinary red-shift of localized surface plasmon resonance from individual GNPs. By quantitatively counting the fraction of yellow particles with color-coded dark-field optical microscopy, the trace amounts of PPi in solution can be accurately quantified. The limit-of-detection is as low as 1.49 nM with a linear dynamic range of 0-4.29 µM, which is much lower than the spectroscopic measurements in bulk solution. In artificial urine sample, good recovery efficiency was achieved. As a consequence, the method demonstrated herein will find promising applications for the ultrasensitive detection of target biomolecules under biological milieu in the future.
RESUMEN
Indoor window film samples were collected in buildings during 2014-2015 for the determination of six phthalate diesters (PAEs). Linear regression analysis suggested that the film mass was positively and significantly correlated with the duration of film growth (from 7 to 77 days). PAEs were detected in all window film samples (n = 64). For all the samples with growth days ranged from 7 to 77 days, the median concentrations of total six PAEs (∑6PAEs) in winter and summer window film samples were 9900 ng/m(2) film (2000 µg/g film) and 4700 ng/m(2) film (650 µg/g film), respectively. Among PAEs analyzed, di-2-ethyl-hexyl phthalate (DEHP) was the major compound (71 ± 9.7%), followed by di-n-butyl phthalate (DBP; 20 ± 7.4%) and diisobutyl phthalate (DiBP; 5.1 ± 2.2%). Positive correlations among PAEs suggested their common sources in the window film samples. Room temperature and relative humidity were negatively and significantly correlated with PAEs concentations (in ng/m(2)). Poor ventilation in cold winter in Noreastern China significantly influenced the concentrations of PAEs in window film which suggested higher inhalation exposure dose in winter. The median hazard quotient (HQ) values from PAEs exposure were below 1, suggesting that the intake of PAEs via three exposure pathways was considered as acceptable.
Asunto(s)
Ésteres , Ácidos Ftálicos , China , Dibutil Ftalato , Humanos , Exposición por Inhalación , Estaciones del AñoRESUMEN
In the present study, the non-fat whipped cream analogue was formulated by the combination of soy protein isolate, different polysaccharides and sucrose. Compared with single polysaccharide, the combined polysaccharide showed synergistic effect on formulating the non-fat whipped cream with better properties. The non-fat whipped creams showed high overrun (up to 570 %), excellent drainage stability (no drainage occurred within 120 min) and comparable hardness (up to 1.1 N) than that of control (a commercially dairy whipped cream). Moreover, the non-fat whipped creams were all solid-like (storage modulus > loss modulus) and exhibited outstanding shape retention ability. These properties were greatly affected by the types and ratios of combined polysaccharide. The combination of anionic and neutral polysaccharides was more beneficial for its properties, and the effect depended on the combined ratios. Especially for samples containing gellan gum/guar gum, their appearance only changed slightly after standing for 60 min, and simultaneously showed satisfying sensory acceptability when the combined ratio was 2/3. Therefore, these novel non-fat whipped creams could be popularized as the functional products aiming at specific groups such as diabetes and obesity people in the future.
Asunto(s)
Polisacáridos , Polisacáridos/química , Proteínas de Soja/química , HumanosRESUMEN
This study evaluated the foaming properties, the dynamic adsorption behavior at the air/water (A/W) interface and the foam rheological characteristics of complexes formed by soy protein isolate (SPI) and different charged polysaccharides, including chitosan (CS), guar gum (GUG) and gellan gum (GEG). The results showed that the SPI/CS10 had the highest initial foam volume (26.67 mL), which were 3.89 %, 100.08 % and 70.19 % higher than that of single SPI, SPI/GUG and SPI/GEG complexes, respectively. Moreover, three charged polysaccharides could all significantly improve the foam stability of complexes. Among them, foams stabilized by SPI/GEG10 were the most stable that the foam volume slightly changed (approximately 1 mL) and no drainage occurred throughout the whole recording process. The interfacial behavior analysis showed that SPI/CS10 had higher diffusion (Kdiff) and rearrangement rate (KR) but lower penetration rate (KP) at the A/W interface compared with single SPI, while SPI/GUG10 and all SPI/GEG complexes showed higher KR and KP but lower Kdiff. In addition, SPI/CS10 was beneficial to concurrently enhance the elastic strength and solid-like behavior of foam system, while all SPI/GEG complexes could improve the elastic strength of foam system but was not conducive to the solid-like behavior.
Asunto(s)
Aire , Polisacáridos , Reología , Proteínas de Soja , Agua , Proteínas de Soja/química , Agua/química , Polisacáridos/química , Gomas de Plantas/química , Galactanos/química , Polisacáridos Bacterianos/química , Quitosano/química , Adsorción , Mananos/químicaRESUMEN
Interactions among multi-component play a critical role in modulating the foaming properties of aerated foods. This study evaluated the mechanisms of synergistic improvement of gellan gum (GEG) and guar gum (GUG) on the foaming properties of soy protein isolate (SPI)-based complex. The results showed that the GEG/GUG ratio was closely related to the intermolecular interactions of SPI-based ternary complex and the dynamical changing of its foaming properties. The SPI/GEG/GUG ternary complex with a GEG/GUG ratio of 2/3 exhibited the highest foamability (195 %) and comparable foam stability (99.17 %), which were 32.95 % and 2.99 % higher than that of SPI/GEG binary complex. At this ratio, GUG promoted the interactions between SPI and GEG, and bound to complex's surface through hydrogen bonding, resulting in the increase of particle size and surface charge, and the decrease of surface hydrophobicity. Although this reduced the diffusion of complex onto the air/water interface, it increased permeation rate and molecular rearrangement behavior, which were the potential mechanisms to improve the foaming properties. Additionally, the synergistic effect of GEG and GUG also enhanced the elastic strength and solid characteristics of foam systems. This study provided a theoretical guidance for the targeted modulation of foaming properties of multi-component aerated foods.
Asunto(s)
Galactanos , Mananos , Gomas de Plantas , Polisacáridos Bacterianos , Proteínas de Soja , Gomas de Plantas/química , Galactanos/química , Proteínas de Soja/química , Mananos/química , Polisacáridos Bacterianos/química , Interacciones Hidrofóbicas e Hidrofílicas , Tamaño de la Partícula , Enlace de HidrógenoRESUMEN
Protein glycosylation is an important post-translational modification and glycoproteins are associated with many crucial metabolic progresses of life. In order to detect glycoproteins sensitively, we propose a gold nanoparticles (GNPs) enumeration method based on boronate affinity sandwich system, which is constructed between the boronic acid polymer functionalized magnetic nanoparticles (Fe3O4@MPS@VPBA NPs) and 4-mercaptophenylboronic acid modified GNPs (GNPs-MPBA) by the targeted glycoproteins as the linker. Therefore, the sandwich complex is formed, resulting in the decrease of GNPs-MPBA counts in the solution. Based on the dark-field microscope (DFM) imaging technique, the sensitive GNPs enumeration assay is developed for glycoproteins quantitation. Immunoglobulin (IgG), as one of the important glycoproteins, is introduced to evaluate the proposed method. A low detection limit of 1.22 ng mL-1 for IgG analysis is obtained. The result indicates that the proposed GNPs enumeration method offers a simple, effective, label-free and highly sensitive strategy without signal amplification. It also possesses great potential for various target molecules determination at the single-particle level in the future.
Asunto(s)
Técnicas Biosensibles , Glicoproteínas/análisis , Oro/química , Nanopartículas del Metal/química , Microscopía Electrónica de Transmisión , Tamaño de la Partícula , Espectroscopía Infrarroja por Transformada de Fourier , Propiedades de SuperficieRESUMEN
Acetylcholinesterase (AChE) plays a vital role in nerve conduction through rapidly hydrolyzing the neurotransmitter acetylcholine (ACh) and is correlated with Alzheimer's disease. In this work, a label-free single-particle enumeration (SPE) method for the quantitative detection of acetylcholinesterase (AChE) activity is developed. The design is based on the fluorescence resonance energy transfer (FRET) between fluorescent conjugated polymer nanoparticles (FCPNPs) and MnO2 nanosheets. The fluorescence of FCPNPs can be effectively quenched by MnO2 nanosheets via hydrogen bonding interaction. In the presence of acetylcholinesterase (AChE), acetylthiocholine (ATCh) could be hydrolyzed to thiocholine (TCh), which can reduce MnO2 to Mn2+ and trigger the decomposition of MnO2 nanosheets. As a result, the fluorescence of FCPNPs is restored. Taking advantage of the superior brightness and stable fluorescence emission from individual FCPNPs, the accurate quantification of AChE is achieved by statistically counting the fluorescent particles on the glass slide surface. A linear range from 5 to 1600 µU mL-1 is obtained for AChE assay and the limit-of-detection (LOD) is 1.02 µU mL-1, which is far below the spectroscopic measurements in bulk solution. In the human serum sample, satisfactory recovery efficiencies are determined in a range of 91.0%-103.0%. Furthermore, pesticide carbaryl as an inhibitor of AChE activity was detected. The LOD is 1.12 pg mL-1 with linear responses ranging from 5 to 300 pg mL-1, which demonstrates the feasibility of this approach for AChE inhibitor screening. As a consequence, the label-free SPE-based method affords a promising platform for the sensitive detection of target molecules in the future.