Effect of Phenol and Alkylamide Interaction on α-Glucosidase Inhibition and Cellular Antioxidant Activity during In Vitro Digestion: Using Szechuan Pepper (Zanthoxylum genus) as a Model.

Although recent evidence indicated significant phenol and alkylamide interaction in aqueous solutions, the gastrointestinal digestion influence of the combination remains unclear. This study aims to investigate phenol and alkylamide interaction during in vitro digestion, focusing on bioaccessibility and bioactivity, including α-glucosidase inhibition and cellular antioxidant activity. Additionally, the structural mechanism of phenol and alkylamide interaction during in vitro digestion was explored. The results indicated that the presence of phenols and alkylamides significantly increased or decreased their respective bioaccessibility, depending on the Zanthoxylum varieties. Furthermore, although antagonistic phenol/alkylamide interaction was evident during α-glucosidase inhibition, cellular oxidative stress alleviation, and antioxidant gene transcription upregulation, this effect weakened gradually as digestion progressed. Glycoside bond cleavage and the methylation of phenols as well as alkylamide isomerization and addition were observed during digestion, modifying the hydrogen bonding sites and interaction behavior. This study provided insights into the phenol/alkylamide interaction in the gastrointestinal tract.

Recent advances in electrochemical cell-based biosensors for food analysis: Strategies for sensor construction.

Owing to their advantages such as great specificity, sensitivity, rapidity, and possibility of noninvasive and real-time monitoring, electrochemical cell-based biosensors (ECBBs) have been a powerful tool for food analysis encompassing the areas of nutrition, flavor, and safety. Notably, the distinctive biological relevance of ECBBs enables them to mimic physiological environments and reflect cellular behaviors, leading to valuable insights into the biological function of target components in food. Compared with previous reviews, this review fills the current gap in the narrative of ECBB construction strategies. The review commences by providing an overview of the materials and configuration of ECBBs, including cell types, cell immobilization strategies, electrode modification materials, and electrochemical sensing types. Subsequently, a detailed discussion is presented on the fabrication strategies of ECBBs in food analysis applications, which are categorized based on distinct signal sources. Lastly, we summarize the merits, drawbacks, and application scope of these diverse strategies, and discuss the current challenges and future perspectives of ECBBs. Consequently, this review provides guidance for the design of ECBBs with specific functions and promotes the application of ECBBs in food analysis.

A cell-based electrochemical taste sensor for detection of Hydroxy-α-sanshool.

The Transient Receptor Potential Vanilloid 1 (TRPV1) has been identified as a suitable candidate for a spicy taste (Zanthoxylum plant) sensor. In this study, we investigated the response of TRPV1 expressed on human HepG2 cell membranes following stimulation with Hydroxy-α-sanshool. A three-dimensional (3D) cell-based electrochemical sensor was fabricated by layering cells expressing hTRPV1. l-cysteine/AuNFs electrodes were functionalized on indium tin oxide-coated glass (ITO) to enhance the sensor's selectivity and sensitivity. HepG2 cells were encapsulated in sodium alginate/gelatin hydrogel to create a 3D cell cultivation system, which was immobilized on the l-cysteine/AuNFs/ITO to serve as biorecognition elements. Using differential pulse voltammetry (DPV), the developed biosensor was utilized to detect Hydroxy-α-sanshool, a representative substance in Zanthoxylum bungeanum Maxim. The result obtained from DPV was linear with Hydroxy-α-sanshool concentrations ranging from 0 to 70 µmol/L, with a detection limit of 2.23 µmol/L. This biosensor provides a sensitive and novel macroscopic approach for TRPV1 detection.

SERS determination of hydroxy-α-sanshool in spicy hotpot seasoning: The strategy to restrain the interference of capsaicin and its mechanism.

Hydroxy-α-sanshool (α-SOH) is the principal ingredient responsible for the numbing sensation in spicy hotpot. However, utilizing surface-enhanced Raman scattering (SERS) to analyze the α-SOH in hotpot seasoning is challenging due to the significant interference of capsaicin (CAP). Therefore, two schemes were proposed to address CAP interference in hotpot seasoning, namely laccase-catalyzed conversion and metal-organic framework (MOF) interaction. Among them, Fe-BTC MOF exhibited significant anti-interference effect and the underlying mechanism is elucidated. The motion of CAP aromatic ring was constrained by steric hindrance and electrostatic interactions of Fe-BTC. Additionally, the interaction between CAP aromatic ring and conjugated triene group in α-SOH was quenched, enhancing the α-SOH SERS signal. The proposed method had a significant anti-interference effect on α-SOH quantification in the presence of CAP, significantly enhancing the α-SOH SERS signal in a range of 0.85 to 4.00 × 107. The linearity and reproducibility of the proposed hotpot seasoning testing method were also validated.

Cooking-Induced Oxidation and Structural Changes in Chicken Protein: Their Impact on In Vitro Gastrointestinal Digestion and Intestinal Flora Fermentation Characteristics.

Meat digestion and intestinal flora fermentation characteristics are closely related to human dietary health. The present study investigated the effect of different cooking treatments, including boiling, roasting, microwaving, stir-frying, and deep-frying, on the oxidation of chicken protein as well as its structural and digestion characteristics. The results revealed that deep-fried and roasted chicken exhibited a relatively higher degree of protein oxidation, while that of boiled chicken was the lowest (p < 0.05). Both stir-frying and deep-frying led to a greater conversion of the α-helix structure of chicken protein into a ß-sheet structure and resulted in lower protein gastrointestinal digestibility (p < 0.05), whereas roasted chicken exhibited moderate digestibility. Further, the impact of residual undigested chicken protein on the intestinal flora fermentation was assessed. During the fermentation process, roasted chicken generated the highest number of new intestinal flora species (49 species), exhibiting the highest Chao 1 index (356.20) and a relatively low Simpson index (0.88). Its relative abundance of Fusobacterium was the highest (33.33%), while the total production of six short-chain fatty acids was the lowest (50.76 mM). Although stir-fried and deep-fried chicken exhibited lower digestibility, their adverse impact on intestinal flora was not greater than that of roasted chicken. Therefore, roasting is the least recommended method for the daily cooking of chicken. The present work provides practical advice for choosing cooking methods for chicken in daily life, which is useful for human dietary health.

Fabrication of a label-free electrochemical cell-based biosensor for toxicity assessment of thiram.

Thiram has been widely used in agriculture and may invades the food chain, posing a threat to human health. In this research, a label-free electrochemical cell-based biosensor was presented for in vitro toxicity assessment of thiram. HepG2 cells were cultured on poly-l-lysine@gold nano-flowers functionalized indium tin oxide coated glass electrode (PLL@AuNFs/ITO) to serve as biorecognition elements. AuNFs were electrodeposited on ITO to provide an enlarged specific surface area and benefited the output signal amplification. PLL was selected as an effective biocompatible coating material to facilitate cell adhesion and proliferation, thereby realizing one-step recording of electrochemical signals from thiram-treated cells. With the aid of the differential pulse voltammetry method, the fabricated biosensor was applied to assess the cytotoxicity of thiram. Results showed that the cytotoxicity measured by the fabricated biosensor exhibited a linear relationship related to thiram concentration ranging from 5 to 50 µM with a detection limit of 2.23 µM. The IC50 of thiram obtained by the biosensor was 29.5 µM, which was close to that of conventional MTT assay (30.8 µM). The effects of thiram on HepG2 cells were also investigated via SEM and flow cytometry. Meanwhile, the proposed biosensor was used to evaluate the toxicity of thiram in fruit samples. Results indicated that the toxicity of thiram cannot be ignored even at a low residual concentration in food (≤5 mg/kg). In conclusion, the developed sensor showed excellent sensitivity, stability, and reliability, which provided a great capacity for the convenient toxicity evaluation of thiram residue in food.

A cell-based electrochemical sensor for assessing immunomodulatory effects by atrazine and its metabolites.

A three-dimensional (3D) cell-based electrochemical sensor was developed to evaluate the immunomodulatory effects of atrazine and its metabolites by monitoring nitric oxide (NO) release. Nafion/Fe(III) meso-tetra (4-carboxyphenyl) porphyrin/reduced graphene oxide (Nafion/FeTCP@RGO) was functionalized on the screen-printed carbon electrode (SPCE) as a working electrode to enhance the NO selectivity and sensitivity. RAW264.7 cells encapsulated in a gelatin methacrylate (GelMA) hydrogel, forming a 3D cell culture system, were immobilized on the Nafion/FeTCP@RGO/SPCE to serve as biorecognition elements. The proposed sensor presented the induced NO expression as a distinct single differential pulse voltammetric (DPV) anodic peak in the selected physiological lipopolysaccharide (LPS) concentration range (0.01-2000 ng/mL). Furthermore, the peak current intensity of the NO was linearly correlated with the LPS concentration logarithm, which was further validated using a Griess reagent kit. The proposed sensor was utilized in optimized LPS-induction conditions to compare the immunosuppression effect of atrazine with its main metabolites (i.e. desethylatrazine (DEA), deisopropylatrazine (DIA), and diaminochlorotriazine (DACT)). The results demonstrated that the order of 30% inhibitory concentrations was atrazine (25.71 ± 1.08 µg/mL) < DEA (48.63 ± 2.17 µg/mL) ≤ DACT (49.11 ± 1.98 µg/mL) ≤ DIA (52.36 ± 2.34 µg/mL). Overall, this work provided a potential in vitro approach to evaluate the immunotoxicity of pesticides and their metabolites.

A signal on-off ratiometric electrochemical sensor coupled with a molecular imprinted polymer for selective and stable determination of imidacloprid.

This work presents a signal on-off ratiometric electrochemical sensor coupled with a molecular imprinted polymer (MIP) for imidacloprid (IMI) determination. The ratiometric strategy corrects the detection results by setting 6-(Ferrocenyl)hexanethiol (FcHT) as an internal reference. The MIP membrane, as a molecular recognition receptor, has a three-dimensional structure and is complementary in the shape and chemical functionality to the template. The combination of a ratiometric strategy and MIP improves the sensitivity and selectivity of the sensors. The experimental parameters, such as monomer/template ratio, electropolymerization cycle and adsorption time, are optimized to improve the IMI sensing performance of the as-fabricated electrode. Under optimal electrochemical conditions, the proposed sensor is used to detect IMI in laboratory and real samples. In a range as wide as 5 × 10-7-1 × 10-4 mol L-1, a linear relationship is well established, and the value of the correlation coefficient is 0.9984. The limit of detection (LOD, S/N = 3) is well defined as 4.7 × 10-8 mol L-1. The recovery rates range from 97.4% to 103.5% while testing real samples. The fabricated electrode shows superior anti-interference ability for IMI determination in the presence of analogs at a higher level.

A nitrile-mediated aptasensor for optical anti-interference detection of acetamiprid in apple juice by surface-enhanced Raman scattering.

Currently, the detection of pesticide is critical for food safety assurance, but it is still challenging due to the presence of biological interferents from complex food matrix. In this study, we developed an optical anti-interference surface-enhanced Raman scattering (SERS) aptasensor system for trace detection of acetamiprid. 4-(Mercaptomethyl) benzonitrile (MMBN) containing CN bond was used as Raman tag to provide a sharp peak (2227 cm-1) in the Raman-silent spectral window (1800-2800 cm-1) where no Raman signal existed for most of molecules. Gold nanoparticles (AuNPs) bonded with polyadenine (polyA)-mediated aptamer and Raman tag (MMBN-AuNPs-aptamer) was synthesized as Raman probe, while the complementary DNA (cDNA) conjugated with AgNPs-decorated silicon wafer (AgNPs@Si) was used as SERS substrate. As acetamiprid molecule could specifically combine with aptamer, preventing the formation of MMBN-AuNPs-aptamer-cDNA-AgNPs@Si (expressed as "Au-AgNPs@Si") hybrid through DNA sequence linking, Raman signal intensities of MMBN in Au-AgNPs@Si decreased when the concentration of acetamiprid increased. Under the optimum assay condition, the proposed method displayed a linear response for acetamiprid detection in the range of 25-250 nM with a low detection limit of 6.8 nM. Finally, the developed aptasensor was successfully used to determine acetamiprid content in apple juice. Accordingly, this novel anti-interference SERS aptasensor could be a promising acetamiprid sensor for food safety assurance.