[Determination of 80 pesticide residues in milk using NH_2-MWNTs by liquid chromatography-time of flight mass spectrometry].

OBJECTIVE: A method for the determination of 80 pesticide residues in milk by liquid chromatography-time-of-flight mass spectrometry(LC-QTof-MS) was developed. METHODS: The target compounds in milk were extracted with acetonitrile-methanol(9∶1, V/V) containing 1% acetic acid, and purified by aminated multi-walled carbon nanotubes(NH_2-MWNTs). The chromatographic column was Waters Acquity UPLC BEH C_(18 )(100 mm×2.1 mm, 1.7 µm). The 80 pesticides were detected by liquid chromatography-time-of-flight mass spectrometry and quantified using an external standard method by matrix matched calibration curve. RESULTS: The purification method showed a good linearity(r~2≥ 0.99) over the concentration range from 5 to 100 µg/L for the 80 pesticides in this study. The limits of detection(LODs) and quantification(LOQs) of the 80 pesticides in milk ranged from 0.01 to 0.50 µg/L and 0.03 to 1.50 µg/L, respectively. The mean recoveries of the three spiked levels ranged from 71.5% to 116.9% with the relative standard deviation ranging from 1.2% to 18.1%, indicating that the accuracy and precision of the method were good. Among the milk samples, no residues of the 80 pesticides in this study were found after screening. CONCLUSION: The method has good linearity, good sensitivity, accuracy and precision and is suitable for the simultaneous and rapid determination of 80 pesticide residues in milk.

Urinary analysis reveals high Alternaria mycotoxins exposure in the general population from Beijing, China.

Alternaria mycotoxins are of concern due to its adverse health effect, they affect various cereal crops and grain-based food along with modified forms that contribute to overall exposure. This study aimed to determine the frequency and level of exposure to Alternaria mycotoxins (tenuazonic acid, TeA; alternariol, AOH; alternariol monomethyl ether, AME; tentoxin, TEN; and altenuene, ALT) in human urine from Beijing adults. A total of 2212 urine samples were collected and analyzed for five mycotoxins using LC-ESI-MS/MS. More than 98% of the samples had at least one Alternaria mycotoxin detected. Among the mycotoxins, AME had the highest detection rate (96.0%), followed by TeA (70.5%). The calculated average daily intake values of AME (12.5 ng/kg b.w.) was 5 times the TTC value (2.5 ng/kg b.w.) set by the EFSA, indicating the potential health risks associated with mycotoxins. Immediate attention and subsequent actions should be taken to identify the sources of mycotoxins and the corresponding exposure pathways to humans in the investigated regions.

AIM2 promotes non-small-cell lung cancer cell growth through inflammasome-dependent pathway.

The human absent in melanoma 2 (AIM2) is considered as a DNA recognizer. AIM2 has been described as a tumor suppressor gene in the early years. But recent studies suggested that it functions as an oncogene in several cancers. However, its roles in non-small-cell lung cancer (NSCLC) remain unclear. Here we reported that AIM2 highly expressed in NSCLC cells and exhibited a tumor-promoting property both in vitro and in vivo. Besides, AIM2 short hairpin RNA (shRNA)-mediated suppression of cell proliferation was triggered by the accumulation of cells at the G2/M phase. Knockdown of AIM2 reduced the inflammasome formation, while overexpression of AIM2 or stimulation by poly(dA:dT) induced the inflammasome formation. Interestingly, blockade of the inflammasome by caspase-1 inhibitor VX-765 or ASC small interfering RNA (siRNA) abolished the effects brought by AIM2 shRNA and AIM2 plasmid. In summary, our results revealed that AIM2 functioned as an oncogene in NSCLC in an inflammasome-dependent way.

Absent in melanoma 2 suppresses epithelial-mesenchymal transition via Akt and inflammasome pathways in human colorectal cancer cells.

Absent in melanoma 2 (AIM2) is a critical component in natural immunity system and is closely related to cancer initiation and development. It has been shown that AIM2 inhibited colorectal cancer (CRC) development and cell proliferation. It remains unresolved how AIM2 acts on CRC metastasis. In this study, we assessed migration, invasion ability, and epithelial-mesenchymal transition (EMT) program upon AIM2 overexpression or knockdown in human CRC cells. Transwell assay demonstrated that upregulation of AIM2 reduced cell migration and invasion. Epithelial marker E-cadherin was augmented and mesenchymal markers vimentin, as well as Snail, were examined decreased by Western blot, real-time polymerase chain reaction, and immunofluorescence. Correspondingly, knockdown of AIM2 led to a reverse consequence. In addition, AIM2 regulated Akt phosphorylation and effects of AIM2 on cell invasion and EMT were recovered after administration of Akt inhibitor, suggesting that AIM2 suppressed EMT dependent on Akt pathway. In addition, caspase-1 inhibitor exposure indicated that AIM2 abrogated EMT through the inflammasome pathway as well. In summary, AIM2 suppressed EMT via Akt and inflammasome pathways in human CRC cells.

Transformation of sulfamethazine during the chlorination disinfection process: Transformation, kinetics, and toxicology assessment.

Various disinfection byproducts (DBPs) form during the process of chlorination disinfection, posing potential threats to drinking water safety and human health. Sulfamethazine (SMT), the most commonly used and frequently detected veterinary antibiotic, was investigated in detail with regard to its transformation and kinetics in reactions with free available chlorine (FAC). Using liquid chromatography coupled to quadrupole time-of-flight tandem mass spectrometry, several DBPs were identified based on different confidence levels, and a variety of reaction types, including desulfonation, S-N cleavage, hydroxylation, and chlorine substitution, were proposed. The kinetic experiments indicated that the reaction rate was FAC- and pH-dependent, and SMT exhibits low reactivity toward FAC in alkaline conditions. The DBPs exhibited a much higher acute toxicity than SMT, as estimated by quantitative structure activity relationship models. More importantly, we observed that the FAC-treated SMT reaction solution might increase the genotoxic potential due to the generation of DBPs. This investigation provides substantial new details related to the transformation of SMT in the chlorination disinfection process.

Uptake, depuration and bioconcentration of bisphenol AF (BPAF) in whole-body and tissues of zebrafish (Danio rerio).

Bisphenol AF (BPAF) is an analog of Bisphenol A (BPA) and is widely used as a raw material in the plastics industry. However, an understanding of the potential risks posed by BPAF in the aquatic environment is lacking. The bioconcentration factor (BCF) is a measure used to assess the secondary poisoning potential as well as risks to human health. In this work we measured the accumulation and elimination of BPAF in the whole-body and in liver, muscle and gonad tissues of zebrafish. BPAF uptake was relatively rapid with equilibrium concentrations reached after 24-72h of exposure. We observed gender differences both in whole-body and in tissue accumulation. Muscle was the primary BPAF storage tissue during the uptake phase in this study. In the elimination phase, BPAF concentrations declined rapidly during depuration, especially during the initial 2h, and the rate of elimination in males was faster than females from the whole-body and from tissues. The appearance of BPAF glucuronide (BPAF-G) at the start of the uptake phase indicated the rapid biotransformation of BPAF to BPAF-G in vivo. The high lipid content of female gonad could act to delay the diffusion of the xenobiotic within the body in a contaminated environment, but it also acts to delay xenobiotic elimination from the body.

Chronically socially isolated mice exhibit depressive-like behavior regulated by the gut microbiota.

Objectives: Chronic loneliness is a widespread issue, and the gut-brain axis is known to be crucial in facilitating communication between the gut and brain. However, the precise mechanism by which chronic loneliness affects the gut-brain axis remains uncertain. Methods: Fourteen 55-week-old Balb/c mice were used in the experiment, with seven mice being randomly assigned to the chronic social isolation (CSI) group. The CSI group mice underwent 12 weeks of isolation to simulate the psychiatric state of a population in prolonged social isolation. The mental state of the CSI mice was assessed through animal behavior analysis, while plasma cytokines were measured using ELISA. Additionally, the composition of the gut microbiota was analyzed using 16S rRNA sequencing, and the metabolite composition of the intestinal contents was examined using nontargeted metabolomics. The Student-T test was used to determine significant mean differences. Results: Mice that were exposed to the CSI exhibited increased immobility time lengths in forced swimming and hanging tail experiments, and decreased movement lengths and number of times traversing the intermediate region, compared to control mice. Additionally, CSI decreased the abundance of the probiotics Ruminococcaceae, Akkermansiaceae, and Christensenellaceae. Additionally, CSI reduced the production of the metabolites oleamide and tryptophan. Furthermore, IL-1ß, IL-4, and IL-6 were significantly increased, while TNF-α was significantly decreased. Conclusion: CSI induces a dysbiotic gut microbiota and the production of neurorelated metabolites, which in turn increase inflammatory responses and result in depressive behaviors in CSI mice. Therefore, these findings suggest that the gut microbiota may serve as a target for the treatment of long-term social isolation-induced mental disorders.

The use of cell and larval assays to identify target genes for RNA interference-meditated control of the Australian sheep blowfly (Lucilia cuprina).

BACKGROUND: Flystrike, primarily caused by Lucilia cuprina, is a major health and welfare issue for sheep wool industries. Current chemical-based controls can have limited effectiveness due to the emergence of resistance in the parasite. RNA interference (RNAi), which uses double-stranded RNA (dsRNA) as a trigger molecule, has been successfully investigated for the development of innovative pest control strategies. Although RNAi offers great potential, the efficient identification, selection of target genes and delivery of dsRNA represent challenges to be overcome for the successful application of RNAi for control of L. cuprina. RESULTS: A primary L. cuprina (blowfly) embryo cell line (BFEC) was established and confirmed as being derived from L. cuprina eggs by PCR and amplicon sequencing. The BFECs were successfully transfected with plasmids and messenger RNA (mRNA) expressing fluorescent reporter proteins and dsRNA using lipid-based transfection reagents. The transfection of dsRNA into BEFC in this study suggested decreased mRNA levels of target gene expression, which suggested RNAi-mediated knockdown. Three of the dsRNAs identified in this study resulted in reductions of in target gene mRNA levels in BFEC and loss of biological fitness by L. cuprina larvae in a feeding bioassay. CONCLUSION: This study confirms that the novel BFEC cell line can be used to improve the efficacy of dsRNA-mediated screening to accelerate the identification of potential target genes in the development of RNAi mediated control approaches for L. cuprina. The research models established in this study are encouraging with respect to the use of RNAi as a blowfly control method, however further improvement and validation are required for field applicationsnot prefect, and could be ongoing developing. © 2024 Society of Chemical Industry.

A SILAC-based accurate quantification of shrimp allergen tropomyosin in complex food matrices using UPLC-MS/MS.

The carryover of trace allergens in complex food matrices poses challenges for detection techniques. Here, we demonstrate an accurate UPLC-MS/MS quantification assay for the shrimp allergen tropomyosin with a full-length isotope-labelled recombinant tropomyosin (TM-I) internal standard in complex food matrices. The TM-I, expressed based on the SILAC technique, exhibited a high isotope labelling ratio (>99%), purity, and alignment with the natural sequence. This method determined the tropomyosin ranging from 0.2 to 100 ng/mL. Mean recoveries ranged from 89 to 116%, with intra- and inter-day RSDs below 12%, for three signature peptides across three types of commercially processed food matrices. The limits of quantitation were 1 µg/g in pop food and sauce, and 10 µg/g in surimi product, respectively. This study supports the use of recombinant full-length isotope-labelled proteins rather than stable-isotope labelling peptides as internal standards to achieve more accurate quantitation of food allergens as the digestion error is corrected.

[An on-line solid phase extraction column switching liquid chromatography tandem mass spectrometry system to analyze bisphenol A and nonylphenol in drinking water].

OBJECTIVE: To develop a method for bisphenol A (BPA) and nonylphenol (NP) analysis in drinking water by on-line solid phase extraction (SPE) column switching liquid chromatography tandem mass spectrometry. METHOD: Target compounds were concentrated and purified by a Waters Direct Connect HP XBridge C18 column, with average particle size 10 microm. The mobile phase of SPE extraction column was methanol/water at a flow-rate of 2.0 ml/min. The analytical column was Waters BEH C18 column (2.1 mm x 50 mm, 1.7 microm) and the mobile phase was methanol/0.1% ammonia water at a flow-rate of 0.4 ml/min. BPA and NP were analyzed by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) in the negative ion mode with multiple reactions monitoring (MRM). RESULTS: The whole run time of an individual sample was about 10 min. The linear calibration curves covered the range of 5 - 1000 ng/L and 10 - 5000 ng/L for BPA and NP, respectively, while the correlation coefficients (r2) were above 0.999. The quantification limits of the method were 5 and 10 ng/L for BPA and NP, respectively. Recovery test was performed at three fortification levels, mean recoveries ranged from 86.6% to 105% with acceptable coefficients of variation (3.11% - 18.2%, n = 6). The inter-day precisions were less than 13.2%. CONCLUSION: The established On-line SPE LC-MS/MS method needs no complicated sample pretreatment step, which is significantly time-saving. The analytical procedure is rapid, simple, and high-sensitive. It can be applied to analysis BPA and NP in drinking water.

Continuous exposure to bisphenol S increases the accumulation of endogenous metabolic toxicants by obstructing the glucuronic acid pathway.

Uridine diphosphate glucuronic acid (UDPGA) is an essential substrate in the glucuronidation of exogenous and endogenous lipophilic compounds via the liver glucuronic acid pathway, and its synthesis depends on glucose and energy in the body. Bisphenol S (BPS), as a lipophilic environmental pollutant, has been widely utilized in the manufacturing of daily necessities. The biological effect of BPS in interference with liver energy metabolism might affect UDPGA synthesis and the excretion of lipophilic compounds, but this was not clearly revealed. Here, female zebrafish that were exposed to BPS for 35 days exhibited a significant decrease in UDPGA in the liver with significant accumulation of exogenous BPS and endogenous bilirubin in the body. One vital reason may be that the exposure to BPS for 35 days promoted the lipid formation through PPARg signaling and reduced energy levels in the liver, resulting in the decreased raw materials for UDPGA production in glucuronic acid pathway. Meanwhile, transcriptome analysis showed that BPS inhibited the mRNA expression levels of genes related to the glucuronic acid pathway. The accumulation of endogenous and exogenous lipophilic compounds can trigger a variety of toxicological effect. Thus, weakened liver detoxification might be the primary cause of the toxicological effects of lipophilic pollutants.

High-throughput multitarget quantitative assay to profile the whole grain-specific phytochemicals alkylresorcinols, benzoxazinoids and avenanthramides in whole grain and grain-based foods.

Currently, there is a growing interest in using whole grain (WG)-specific phytochemicals to perform WG research, including research on dietary assessment, health mechanisms, and quality control. However, the current approaches used for WG-specific phytochemical analysis cannot simultaneously achieve coverage, specificity, and sensitivity. In the present study, a series of WG-specific phytochemicals (alkylresorcinols (ARs), benzoxazinoids (BXs) and avenanthramides (AVAs)) were identified, and their mass spectrometry (MS) fragmentation mechanism was studied by TOF MS. Based on diagnostic fragmentation ions and retention time prediction models, a LC-MS/MS method was developed. Through this method, 56 ARs, 13 BXs, and 19 AVAs in WGs and grain-based foods were quantified for the first time. This method was validated and yielded excellent specificity, high sensitivity and negligible matrix effects. Finally, we established WG-specific phytochemical fingerprints in a variety of WG and grain-based foods. This method can be used for WG quality control and WG precision nutrition research.

Multi-immunoaffinity column for the simultaneous analysis of bisphenol A and its analogues in Chinese foods by liquid chromatography tandem mass spectrometry.

Bisphenol A (BPA) and its four analogues have been receiving considerable attention owing to their potential endocrine disrupting effects. The European Food Safety Authority has proposed 0.04 ng/kg·body weight/day of thetemporary tolerable daily intake for BPA. Therefore, a more sensitive analytical method was urgently needed for the necessity of the risk reassessment of bisphenols (BPs). The matrix effect of Chinese foods is a challenge for the analysis of ultra-trace analytes due to the presence of various spices. A multi-immunoaffinity column (mIAC) was prepared for the purification of BPA, BPB, BPF, BPS, and BPAF in Chinese foods following ultra-high-performance liquid chromatography tandem mass spectrometry detection (UHLPC-MS/MS). The recoveries of each of BPs were ranged from 84.6% to 116.7%, and the intra-day precision and inter-day precision were ranged from 1.6% to 12.4%, and from 4.1% to 14.0%, respectively. This is the first report on the mIACs for simultaneous clean-up and analysis of BPs in complex Chinese foods.

Occurrence and migration of organophosphite and organophosphate esters into food simulants from single-use food packaging in China.

Organophosphite antioxidants (OPAs) and organophosphate esters (OPEs) are used as additives in food packaging. Because these chemicals have been found in various foods, they have caused increasing concern about potential health risks through food intake. Little information is available about the migration behaviors of OPAs and OPEs from single-use food packaging into food. In the present study, four OPAs and 23 OPEs were analyzed in paper and plastic single-use food packaging (n = 312), which are widely used for take-out food in China. The total concentrations of OPAs and OPEs in the packaging samples were 1966 and 189 ng/g, respectively. Tris (2,4-di-tert-butylphenyl) phosphite (AO168) was the dominant compound. OPAs and OPEs were present at higher concentrations in the plastic packaging than in the paper packaging. In a migration test, four OPAs and 15 OPEs were found in food simulants (4% acetic acid, 10% ethanol, and hexane). Higher levels of individual and total OPAs were found in hexane than the other food simulants, especially for AO168 migration from plastic packaging. The amounts of OPEs in the food simulants increased from the aqueous simulants (4% acetic acid and 10% ethanol) to the fatty food simulant (hexane). The migration efficiencies of the OPAs were higher than those of the OPEs. Preliminary calculations suggest that dietary exposure to OPAs and OPEs because of migration will be low for the population in China.

Characterization of the molecular properties and allergenicity (IgE-binding capacity) of ß-lactoglobulin by heat treatment using asymmetric-flow field-flow fractionation and ultra-performance liquid chromatography quadrupole time of flight mass chromatography.

In this study, the heat product (90 °C, 10 min) of ß-lactoglobulin (ß-LG) was analyzed by asymmetric-flow field-flow fractionation (AF4) to observe the effect of heat treatment. The changes in molar mass (M) and molar size induced by heat treatment were characterized by AF4, and changes in molar shape were observed by transmission electron microscopy (TEM). The results showed that ß-LG dissociated and aggregated into four fractions with different M values, sizes, and shapes after heat treatment. The vast aggregations with the highest allergenicity (IgE-binding capacity) might enhance the allergenicity of ß-LG. However, the number of characterized epitope peptides was decreased due to heat treatment. The above results provide some references for related studies of ß-LG and its allergenicity. Further separation and characterization of the high-allergenicity fractions and peptides will help to eliminate allergens in dairy products and reduce the occurrence of allergic reactions.

Occurrence and Estimated Daily Intake of Cortisone and Cortisol in Aquatic Food from China TDS.

Glucocorticoids (GCs) widely exist in animal food including aquatic food. This study aimed to survey the occurrences of cortisone and cortisol in aquatic food and the estimated daily intake (EDI) of cortisone and cortisol due to different habits of aquatic food consumption. The mean levels of cortisone and cortisol in freshwater fish purchased from market were 14.59 µg/kg and 69.15 µg/kg, respectively, which were markedly higher than the levels in marine fish. A test using Zebrafish was performed to compare the concentration of GCs by different killing methods. The results suggested that physically traumatic killing methods are one of the reasons why the levels of GCs in freshwater fish were higher than those in marine fish. The concentrations of cortisone and cortisol in composite aquatic food samples from 12 provincial districts of the fourth China Total Diet Study (TDS) were 0.72~15.75 µg/kg and 4.90~66.13 µg/kg, respectively, which were positively correlated with the distance from the coastline. Further, the correlation coefficient between the levels of cortisone and cortisol in aquatic food and the percentages of freshwater fish consumption were 0.758 (p < 0.01) and 0.908 (p < 0.01), respectively. There was a significant positive correlation between the levels of cortisone and cortisol in aquatic food in the fourth TDS and the percentages of freshwater fish consumption. The calculated average EDIs of cortisone and cortisol from aquatic food in the fourth TDS were 0.16 µg/d and 0.72 µg/d, respectively.

Combination of magnetic beads extraction and ultraperformance liquid chromatography tandem mass spectrometry detection for the clinical diagnosis of allergies.

The total amount of immunoglobulin E (IgE) in human serum is an important parameter in diagnosing allergies. To reduce the false diagnosis of allergies and better assist in therapy, clinical studies can be performed to obtain accurate and reliable measurements of IgE. A magnetic beads (MBs)-based ultraperformance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for total IgE measurement and the diagnosis of food allergies in serum was developed in this study. First, IgE was extracted by MBs coupled with anti-IgE antibody from serum. The extracted IgE was quantified by a specific signal peptide after digestion. A spiked linear IgE concentration ranging from 400 to 5000 ng mL-1 was used for quantification. The limits of detection and quantification were 400 ng mL-1 and 800 ng mL-1, respectively, for the developed method. Additionally, the combined capacity of the extracted IgE with different allergic proteins was evaluated by a binding experiment in vitro. The combining capacity of IgE with different allergens was used to speculate the kind of allergens that induce allergies in patients. Overall, a new method was developed that could be used to quantify the amount of IgE and simultaneously diagnose which allergen causes an allergic reaction, and this method may provide a powerful new tool in the clinical detection of allergies. Moreover, the developed method was applied to analyze IgE in four samples of patient serum and four serum samples from healthy individuals.

The amino - functionalized magnetic graphene oxide combined with graphite furnace atomic absorption spectrometry for determination of trace inorganic arsenic species in water samples.

A novel adsorbent of magnetic graphene oxide (GO) chemically modified by cysteamine hydrochloride (Fe3O4@SiO2/GO-NH2) through thiol-ene click chemistry reaction was synthesized. The prepared Fe3O4@SiO2/GO-NH2 exhibit selective adsorption to As(V) with high adsorption capacity (52.66 mg g-1). Taking Fe3O4@SiO2/GO-NH2 as the adsorbents, a new method of magnetic solid phase extraction (MSPE) combined with graphite furnace atomic absorption spectrometry (GFAAS) was developed in determining trace-level inorganic arsenic species (As(III) and As(V)) in environmental water and bottled water samples. Various experimental parameters affecting the MSPE have been optimized. Under the optimal experimental parameters, the limit of detection of the established method for As(V) was 1.02 ng L-1, the relative standard deviations were 7.9% (intra-day, c = 50 ng L-1, n = 5) and 4.6% (inter-day, c = 50 ng L-1, n = 7), respectively, and the enrichment factor of the method was 392. GBW08666 and GBW08667 (certified reference material) were analyzed to confirm the accuracy of the method, and the results were matched well with the certified values. The established MSPE-GFAAS method was successfully applied in analyzing trace/ultratrace As(III) and As(V) in real water samples.

Effect of chewing betel nut on the gut microbiota of Hainanese.

Betel nut chewing (BNC) is prevalent in South Asia and Southeast Asia. BNC can affect host health by modulating the gut microbiota. The aim of this study is to evaluate the effect of BNC on the gut microbiota of the host. Feces samples were obtained from 34 BNC individuals from Ledong and Lingshui, Hainan, China. The microbiota was analyzed by 16S rRNA gene sequencing. BNC decreased the microbial α-diversity. Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria were the predominant phyla, accounting for 99.35% of the BNC group. The Firmicutes-to-Bacteroidetes ratio was significantly increased in the BNC group compared to a control group. The abundances of the families Aerococcaceae, Neisseriaceae, Moraxellaceae, Porphyromonadaceae, and Planococcaceae were decreased in the BNC/BNC_Male/BNC_Female groups compared to the control group, whereas the abundances of Coriobacteriaceae, Streptococcaceae, Micrococcaceae, Xanthomonadaceae, Coxiellaceae, Nocardioidaceae, Rhodobacteraceae, and Succinivibrionaceae were increased. In general, the gut microbiome profiles suggest that BNC may have positive effects, such as an increase in the abundance of beneficial microbes and a reduction in the abundance of disease-related microbes. However, BNC may also produce an increase in the abundance of disease-related microbes. Therefore, extraction of prebiotic components could increase the beneficial value of betel nut.

UPLC-MS/MS and Network Pharmacology-Based Analysis of Bioactive Anti-Depression Compounds in Betel Nut.

BACKGROUND: Betel nuts have long been used in traditional Chinese medicine. In our study, the bioactive components of betel nut were systematically investigated, and the main components and their target genes in the treatment of depression were predicted. METHODS: The metabolites of the kernels and peels were analyzed with a UPLC-MS/MS system. Mass spectrometry outcomes were annotated by MULTIAQUANT. "Compound-disease targets" were utilized to construct a pharmacology network. RESULTS: A total of 873 metabolites were identified, with a high abundance of flavonoids, alkaloids, and phenols. Moreover, the abundance of flavonoids, alkaloids, and phenols in the kernel was significantly higher than that in the peel. A high abundance of catechin, arginine, and phenylalanine was detected in the kernel, while a high abundance of arginine, arecoline, and aminobutyric acid was detected in the peel. Catechins and cyanoside were the most abundant flavonoids in the kernel and peel, respectively. Arecoline was the most abundant alkaloid. A total of 111 metabolites showed a significant difference between the kernels and peels. The relative abundance of 40 differential metabolites was higher than 100,000, including 14 primary metabolites, 12 flavonoids, 4 phenols, and 4 alkaloids. Among the 40 high abundance metabolites, 20 were higher in the kernel and 20 in the peel. In addition, the enrichment of metabolic pathways found that the kernel and peel of the fruit adopted different metabolic pathways for the synthesis of flavonoids and alkaloids. Network pharmacology prediction showed that 93 metabolites could target 141 depression-related genes. The main components of betel nut intervention in depression were predicted to include L-phenylalanine, protocatechuic acid, okanin, nicotinic acid, L-tyrosine, benzocaine, syringic acid, benzocaine, phloretic acid, cynaroside, and 3,4-dihydroxybenzaldehyde. CONCLUSION: Betel nuts are rich in natural metabolites, and some of these metabolites can participate in the intervention of depression. In addition, the metabolites showed distinct characteristics between the kernel and peel. Therefore, it is necessary to comprehensively and rationally use betel nuts.