This study established a method using gas chromatography-electrostatic field Orbitrap high-resolution mass spectrometry (Orbitrap GC-MS) for the simultaneous determination of 70 organic pollutants across four categories: organochlorine pesticides (OCPs), polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and phthalates (PAEs) in infant cereal-based supplementary foods. Techniques integrating cryogenic centrifugation and lipid and protein adsorbent (LPAS) purification were employed. The precise mass numbers of target compounds were determined by Orbitrap GC-MS in full-scan mode (Full MS), which effectively minimized matrix interferences in these foods. Method validation on rice flour samples demonstrated that the 70 compounds exhibited excellent linearity within their respective mass concentration ranges, with correlation coefficients all exceeding 0.995. The detection limits for this method ranged from 0.10-1.0 μg/kg, while the quantification limits varied from 0.3-3.0 μg/kg, meeting the established detection requirements. The average spike recovery of the 70 compounds at three spiked levels (5, 20, and 100 μg/kg) ranged from 75.3%-119.3%, with relative standard deviations ranging from 1.8%-10.8%. Both inter-day and intra-day precision demonstrated relative standard deviation values below 15%. This method was applied to analyze 100 samples of commercial infant cereal-based supplementary food, revealing the presence of PAEs and PAHs in 12.0% of the samples. Notably, no OCPs or PCBs were detected. The detected concentrations of benzo[a]pyrene (BaP) and dibutyl phthalate (DBP) were (4.2 ± 0.1) μg/kg and (1.8 ± 0.3) μg/kg, respectively. This method is straightforward, highly sensitive, and suitable for the rapid screening and confirmation of 70 organic pollutants in infant cereal-based supplementary foods.
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Inflammation caused by obesity, particularly in adipose tissue and the liver, can lead to insulin resistance (IR) and trigger type 2 diabetes (T2DM). It is crucial to identify therapeutic agents that alleviate IR by reducing inflammation. Here, we report that isobavachromene (IB), a flavonoid derived from Psoralea corylifolia Linn., ameliorates IR in 3T3-L1 adipocytes by inhibiting the MAPK/NF-κB signaling pathway. We first found that IB could promote glucose uptake in 3T3-L1 adipocytes by activating the PI-3K/Akt signaling pathway and was more effective than the positive control sodium orthovanadate at concentrations ranging from 25 to 100 μmol/L. Additionally, IB inhibited RAW264.7 macrophage infiltration into 3T3-L1 adipocytes and suppressed the secretion of inflammatory factors from RAW264.7 macrophages, as well as the phosphorylation levels of key proteins (NF-κB P65, ERK1/2, JNK, and P38) in the NF-κB and MAPK signaling pathways. In summary, IB improves glucose uptake in IR adipocytes, activates the PI-3K/Akt signaling pathway, inhibits the JNK and NF-κB inflammatory signaling pathways, and reduces adipocyte inflammation, thereby improving of IR in 3T3-L1 adipocytes.
Intermittent fasting can benefit breast cancer patients undergoing chemotherapy or immunotherapy. However, it is still uncertain how to select immunotherapy drugs to combine with intermittent fasting. Herein we observed that two cycles of fasting treatment significantly inhibited breast tumor growth and lung tissue metastasis, as well as prolonged overall survival in mice bearing 4T1 and 4T07 breast cancer. During this process, both the immunosuppressive monocytic- (M-) and granulocytic- (G-) myeloid-derived suppressor cell (MDSC) decreased, accompanied by an increase in interleukin (IL) 7R+ and granzyme B+ T cells in the tumor microenvironment. Interestingly, we observed that Ly6Glow G-MDSC sharply decreased after fasting treatment, and the cell surface markers and protein mass spectrometry data showed potential therapeutic targets. Mechanistic investigation revealed that glucose metabolism restriction suppressed the splenic granulocyte-monocyte progenitor and the generation of colony-stimulating factors and IL-6, which both contributed to the accumulation of G-MDSC. On the other hand, glucose metabolism restriction can directly induce the apoptosis of Ly6Glow G-MDSC, but not Ly6Ghigh subsets. In summary, these results suggest that glucose metabolism restriction induced by fasting treatment attenuates the immune-suppressive milieu and enhances the activation of CD3+ T cells, providing potential solutions for enhancing immune-based cancer interventions.
Plant protein beverage adulteration occurs frequently, which may cause health problems for consumers due to the hidden allergens. Hence, a novel method was developed for authentication by UPLC-MS/MS. Almond, peanut, walnut and soybean were hydrolyzed, followed by separation by NanoLC-Triple TOF MS. The obtained fingerprints were identified by ProteinPilotTM combined with Uniprot, and 16 signature peptides were selected. Afterwards, plant protein beverages treated by trypsin hydrolysis were analyzed with UPLC-MS/MS. This method showed a good linear relationship with R2 > 0.99403. The LOQs were 0.015, 0.01, 0.5 and 0.05 g/L for almond, peanut, walnut and soybean, respectively. Mean recoveries ranged from 84.77% to 110.44% with RSDs < 15%. The developed method was successfully applied to the adulteration detection of 31 plant protein beverages to reveal adulteration and false labeling. Conclusively, this method could provide technical support for authentication of plant protein beverages to protect the rights and health of consumers.
Human normal flora is a source of probiotics. The safety characteristics of a specific isolate determine its application in foods or drugs. The food-borne-pathogen antagonist strain Lactobacillus gasseri HMV18 is one of the isolates from normal human f lora. In this work, we assessed the in vitro pH tolerance, bile tolerance, biogenic amine production, mucin utilization, and safety of in vivo administration to mice to evaluate general health, organ-body weight index, organ histopathological change, whether L. gasseri HMV18 can colonize in the gut or modulate the gut microbiota after oral administration. The results suggest that L. gasseri HMV18 can tolerate pH 3 for 2 h, 3% bile for 3 h, biogenic amine negative, mucin usage negative, does not encode verif ied toxins, and cause no visible change in mice’s organs. L. gasseri HMV18 might not colonize in mice’s gut, but can signif icantly affect the structure of gut microbiota. A bibliographical survey suggested that there were as few as 8 opportunistic infection cases from 1984 to 2022 and that the possibility for L. gasseri to cause infection is relatively low. Therefore, this work provides a basis for the foods or drugs application of L. gasseri HMV18 and gives a map of experiments for the safety assessment of probiotics.
Patrinia scabiosaefolia, is used as wild vegetable in China for more than 2 000 years, with a variety of pharmacological activities, including anti-inflammatory, anti-tumor and hypoglycemic. Based on our ongoing research on chemical constituents and hypoglycemic activity of P. scabiosaefolia, 4 lignan compounds, (+)-isolariciresinol (1), 7R,7′R,8S,8′S-(+)-neo-olivil-4-O-β-D-glucopyranoside (2), 4-O-methylcedrusin (3) and patrinian A (4), were isolated and identified. The hypoglycemic activity showed that compounds 2 and 3 could extremely significantly improve insulin resistance at 100 (P < 0.001), 50 (P < 0.001) and 25 µmol/L (P < 0.01) in IR 3T3-L1 cells. While compound 4 only promoted glucose uptake by IR 3T3-L1 cells at 100 µmol/L (P < 0.01). Western blotting experiments showed that compounds 2 and 4 up-regulated the protein expressions of p-IRS, PI-3K, p-AKT and glucose transporter 4 (GLUT4), and promoted the transcription of GLUT4 mRNA. Therefore, the mechanisms of compounds 2 and 4 were presumed to improve IR by activating PI-3K/AKT signaling pathway.
Benzo[a]pyrene (B[a]P) is a food contaminant toxic for cardiovascular diseases. The nuclear translocation of Arylhydrocarbon receptor (AhR) plays an important role in B[a]P-induced oxidative stress and vascular diseases. We confirmed that B[a]P promoted ROS production in vascular smooth muscle cells (VSMCs) in vitro and in vivo, associated with the nuclear translocation of AhR. It is known that phosphorylation inhibits while dephosphorylation of AhR promotes nuclear translocation of AhR. However, from the posttranslational modification level, the mechanism by which B[a]P activates and regulates the nuclear translocation of AhR is unclear. Co-immunoprecipitation results showed that cytoplasmic AhR was phosphorylated before B[a]P stimulation, and switched to O-GlcNAcylation upon B[a]P 1-h stimulation in VSMCs, suggesting there may be a competitively inhibitory relationship between O-GlcNAcylation and phosphorylation of AhR. Next, siRNAs of O-linked N-acetylglucosamine transferase (OGT), O-GlcNAcase (OGA) and OGA inhibitor PUGNAc were used. SiOGT blocks but siOGA and PUGNAc promote B[a]P -dependent AhR nuclear translocation and oxidative stress. Ser11 may be the competitive binding site for phosphorylation and O-GlcNAcylation of AhR. Phosphorylation-mimic variant inhibits but O-GlcNAcylation of AhR promotes AhR nuclear translocation and oxidative stress. Our findings highlight a new perspective for AhR nuclear translocation regulated by the competitive modification between phosphorylation and O-GlcNAcylation.
A new method for screening and identification 420 pesticide residues in fruits and vegetables by ultra-performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UPLC-Q-TOF/MS) were developed. The samples were extracted with acetonitrile/acetic acid (99:1, V/V), and clean-up by SinChERS-Nano (single-step, cheap, effective, rugged, safe, nano) column, determined by UPLC-Q-TOF/MS. The accurate mass database and MS/MS database which contains 420 pesticides were established, the automatic retrieval of detection results was carried on according to the accurate mass, retention time, isotope ratio, ion fragment information, and so on. Method verification was performed on leeks samples. The results showed that 420 pesticides had good linearity in the range of 0.1–100 μg/L, and the correlation coeffificients (R2) was greater than 0.990. The limits of detections (LODs) and limits of quantifications (LOQs) of 420 pesticides were in range of 0.05–2.0 and 0.1–5.0 μg/L, respectively. The average spike recoveries at 3 levels were 70.1% to 119.7%, and the relative standard deviations (RSD) were lower than 20% (n = 6). With this method, a survey of pesticide residues was conducted for 110 samples of 10 different fruits and vegetables, which provided scientific data for ensuring pesticide residue safety of the fruits and vegetables consumed daily by the public. This method was simple, sensitive and accurate, and could be used for rapid screening of 420 pesticide residues in fruits and vegetables.
Flower plants are popular all over the world and important sources of ornamental plants, bioactive molecules and nutrients. Flowers have a wide range of biological activities and beneficial pharmacological effects. Flowers and their active ingredients are becoming more and more popular in the preparation of food, drugs and industrial products. This paper summarizes the active ingredients, pharmacological activities and applications in the pharmaceutical and food industries of flower plants in recent years. In addition, the possible molecular mechanism of pharmacological effects of flower plants were also discussed. 302 active constituents from 55 species of flower plants were summarized, including flavonoids (115), terpenoids (90), phenylpropanoids (20), alkaloids (13), organic acids (27) and others (37). The pharmacological effects of flower plants are very extensive, mainly including antioxidant, anti-inflammatory, anti-tumor, anti-virus, and hypoglycemic. The mechanisms of anti-inflammatory, anti-tumor and hypoglycemic activities present the characteristics of multi-way and multi-target. Because of its rich nutrients, bioactive ingredients and plant essential oils, and its wide sources, flower plants are widely used in food, beverage, cosmetics and drug research. Flower plants also play an important role in pharmaceutical industry, food industry and other fields.
The aim of this work was to develop a liquid chromatography-tandem mass spectrometry method for the determination of milk allergen and egg allergen in food products. Signature peptides GGLEPINFQTAADQAR, VGINYWLAHK, VLVLDTDYK, FFVAPFPEVFGK, and NAVPITPTLNR were confirmed and synthesized as the quantitative peptide of ovalbumin, α-lactalbumin, β-lactoglobulin, αS1-casein and αS2-casein, the relative isotope-labeled internal standards were used in the quantitative analysis. Linear range was in the range of 0.5–5000.0 nmol/L for egg and milk allergen in bread, cake, cookie, rice crust and wheat flour samples with free from egg and milk, the limits of detection of milk allergens and egg allergen were in the range between 0.94 mg/100 g and 56.71 mg/100 g, limits of quantification of milk allergens and egg allergen were in the range between 2.36 mg/100 g and 141.78 mg/100 g. The recoveries ranged from 76.7% to 122.8%, the relative standard deviations were in the range of 1.60%–15.60%. The developed method has been successfully used for the detection of egg and milk allergen in various food samples.