The widespread use of structural analogs to replace the endocrine-disrupting chemicals bisphenol A (BPA) and bisphenol S (BPS) has become a growing concern for both the environment and human health. As a novel BPS derivative, 2,4-bis(phenylsulfonyl)phenol (DBSP) has been frequently detected in environmental samples and human biological matrices; however, the biotransformation and toxicokinetic behavior of DBSP remain uncharacterized. To address these gaps, liquid chromatography coupled with high-resolution mass spectrometry (HRMS) was used in this study to identify DBSP metabolites from human liver microsomes/cytosolic fractions in vitro. For the first time, oxidative metabolites, as well as glucuronide and sulfate conjugates of DBSP, were identified, and a corresponding LC–MS/MS semiquantitative method was established. Further phenotyping assays using recombinant human UGT and SULT isoforms revealed that UGT1A3, UGT1A8, UGT1A9 and UGT1A1 exhibited high glucuronidation activity toward DBSP, while SULT1E1, SULT1A1, and SULT1B1 played major roles in DBSP sulfation. Pharmacokinetic analysis in rats after a single oral dose (100 μg/kg) further underscores the importance of phase II conjugation in DBSP metabolism. Compared with that of BPS, the proportion of free forms of DBSP in plasma was greater (77.0 ± 6.2%), and its elimination half-life (10.7 h) was longer, suggesting potential toxicological risk. These findings provide a critical foundation for future biomonitoring and toxicological assessments of DBSP.

Background:Birth defects have become the leading cause of death among children, placing a heavy burden on society and families. Various elements from the environment may be associated with birth defects in the fetus, but few prospective studies have investigated the relationship between multiple-element exposure in the first trimester and birth defects. Methods:In this study, 271 cases were matched to 542 controls for age (±2 years) and gestational age (±2 weeks). We measured the concentrations of 17 elements in urine samples collected from pregnant women during the first trimester and used conditional logistic regression models to estimate odds ratios (ORs) and 95% confidence intervals (95% CIs). The BKMR model was used to evaluate the correlation between element mixed exposure and birth defects. Results:In the single-metal multivariate model, aluminum (Al), chromium (Cr), manganese (Mn), iron (Fe), nickel (Ni), and zinc (Zn) were negatively associated with total birth defects. Restricted cubic splines revealed linear or nonlinear dose-responsive relationships between either Ni or Zn and the risk of birth defects. BKMR results showed that birth defects showed a trend of first slightly rising and then declining; when the element mixture was above the 50th percentile, it was significantly negatively associated with total birth defects. Ni and Zn showed an obvious trend of slight increase and then rapid decrease, indicating that there was a dose-response relationship between these elements and birth defects. There were interactions between Ni and the other five elements. Conclusion:The mixture of high levels of Al, Cr, Mn, Fe, Ni, and Zn in urine during the first trimester of pregnancy was related to a reduction in total birth defects. In particular, the influence of Ni and Zn on total birth defects should be considered.

Bisphenol S (BPS), considered a "regrettable substitute" for the well-known endocrine disruptor bisphenol A (BPA), has also been demonstrated to pose health risks. An underinvestigated exposure route is the placement of thermal price labels on fresh food packaging. Here, a structure-guided suspect screening strategy was employed to characterize BPS derivatives in 137 supermarket price labels. A total of 26 candidate features were identified by high resolution mass spectrometry, among which 9 BPS derivatives were confirmed by authentic standards, and 4 candidates were identified with a confidence of level 2. DBSP, BPS, BPS-MAE and D-90 were the major derivatives, with maximum concentrations ranging from 7.5 to 40 mg/g. A novel derivative, HDPS was first identified as a component coexisting with DBSP in thermal price labels. The concentrations of DBSP were higher than those of BPS and had a significant negative correlation (r = -0.53, P < 0.001), suggesting an ongoing substitute for BPS. Analysis of matched food samples revealed consistent contamination trends: BPS was detected in 89.0% of samples, followed by DBSP (86.1%), BPS-MAE (60.6%), and BPA (59.8%). High levels of BPS (129.4 ng/g), BPA (124.8 ng/g), DBSP (78.1 ng/g) and BPS-MAE (63.8 ng/g) could be observed in individual cling film-wrapped food items with attached price labels. Dietary intake assessments revealed a median & sum;BPs exposure level of 5.6 ng/kg bw/day, but a level of 92 ng/kg bw/day in high-exposure scenarios. The estimated DBSP, BPS, and BPA exposure levels exceeded the stringent EFSA TDI for BPA. This study provides insight into the potential contamination of fresh food caused by thermal labels, informing policies for preventing "regrettable substitutions".

Disinfection byproducts (DBPs), particularly haloacetic acids (HAAs) and oxyhalides, are of global concern due to their carcinogenicity and occurrence beyond drinking water. Biomonitoring data in human biological matrices remain scarce, especially for emerging iodinated DBPs. This study presents a validated analytical method for simultaneously quantifying 11 HAAs and oxyhalides in serum, urine, and breast milk. This method combines a tailored solid-phase clean-up with ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS) and achieves excellent recoveries of 77-117 %, low method detection limits of 0.01-0.20 mu g l-1, and acceptable matrix effects across all matrices. Applied to 122 biological samples collected in Beijing, this approach revealed several unprecedented findings: (i) mono-iodoacetic acid (MIAA), a highly toxic iodinated DBP, was consistently detected in all three matrices, marking the first such report in human biomonitoring; (ii) chlorate was detected in human urine for the first time, with a mean concentration of 117 mu g l-1, indicating overlooked exposure pathways; and (iii) significant correlations between HAAs and oxyhalides in biological samples suggest shared exposure sources or metabolic processes not previously documented. Serum exhibited the highest HAA concentrations, suggesting its high body burden. Chlorate and perchlorate were frequently detected at high concentrations in urine and breast milk, highlighting possible risks to vulnerable populations. This study addresses critical gaps in methodology and exposure data, providing a robust tool for DBP biomonitoring and key evidence for cumulative risk assessment beyond drinking water.

Biomarkers of food intake (BFIs) for whole grains (WGs) would enable more precise dietary assessments and help investigate WG's health effects. However, no reports have been published on the biomarkers that reflect the intake of WG wheat in the Chinese diet. In an acute, randomized, crossover intervention study performed on 22 Chinese subjects, WG wheat BFI candidates were screened using an LC-HRMS untargeted metabolomics technique. Screening results indicate that alkylresorcinol (AR) metabolites are WG wheat-specific metabolites. These metabolites were systematically characterized by in vitro metabolism reaction, and the matched high-throughput LC-MS/MS-targeted quantitative method was developed. Time-response plots generated via targeted analysis revealed AR oxidation products (AR-OOH) and their sulfate conjugates (AR-OOH-sul) increased rapidly following the WG wheat consumption, which were identified as novel and short-term WG wheat BFIs. Another attractive biomarker was glucuronidated ARs (AR-glu), which can distinguish between WG and refined grain (RG) groups 24 h after WG wheat ingestion and is considered a promising medium- and long-term biomarker. Subsequently an independent dose-response study was performed for 38 volunteers who consumed different WG wheat amounts (0, 25, 50 or 100 g) for further validation. Identified BFIs perform well in classifying participants into WG wheat consumers or non-consumers, and could capture dose-dependent changes with WG intake increased. Overall, this is the first study to discover and validate WG BFIs in the Chinese population via dietary intervention trials, indicating the potential usefulness for WG wheat intake assessments and dietary compliance monitoring. Future work will examine their use in large-scale free-living populations.

Regulatory heterogeneity on the maximum residue levels (MRLs) of hazardous substances for food is identified as a challenge of trade between countries. To balance the import and export trade of milk, a high-throughput determination method was established for hazardous substances with regulatory differences. In this paper, we investigated 462 hazardous substances with different MRLs for food based on different countries’ regulations, involving pesticides, veterinary drugs, and some toxins. A mass database was established for these compounds including the basic information, retention time, collision cross section, parent ion, and product ions with ultra-high performance liquid chromatography ion mobility quadrupole time-of-flight mass spectrometry (UPLC-IM-QTOFMS). After that, the sample preparation for milk, including extraction solvents and purified sorbents, was optimized by selecting 274 hazardous compounds as the representative compounds. Acetonitrile/methanol (9:1, v/v) containing 1% acetic acid was used for extracting, and 50 mg EMR and 50 mg PSA were used for purifying the target substances in milk. Then, the methodology was evaluated by spiking the compounds in real milk. The experiment was conducted by matrix calibration, and the results displayed that most compounds had good linearity within their linear ranges (R2 > 0.99). The recovery ranged from 61.8% to 119.7% at three spiking levels, with RSDs between 1.1% and 20.2%. The limits of quantitation of target compounds in milk ranged from 1 to 10 μg/kg. This could meet the MRL requirements among different countries. All the results demonstrated this determination technology was a fast, sensitive, and accurate method for screening hazardous substance.

Ovalbumin (OVA) is a high-risk allergen with complex tertiary structure in food samples. Here, we developed an accurate UPLC-MS/MS-based assay to improve OVA quantitative performance in processed foods. Full-length isotope-labeled OVA proteins (OVA-I) were synthesized using stable isotope labeling by amino acids in cell culture (SILAC) technique and employed as functional internal standards to ensure similar cleavage sites between internal standards and analytes. Subsequently, OVA and OVA-I fully expose their key cleavage sites by using 10 mol/L urea denaturation to correct errors caused by structural variations in different food matrices, the possible denaturation mechanism was also explored. Three quantitative peptides were selected for OVA detection, achieving 88 %-117 % recoveries with RSDs <9.36 % across three different spiking levels. The LOQs were set as 5 μg/g. OVA risk assessment in Chinese general population exhibited high exposure risks with intake level of most processed foods ranged from 0.35 to 215.66 mg.

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 center dot body weight/day of the temporary 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.

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 (2,4di-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.

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.

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 mu g/g in pop food and sauce, and 10 mu g/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.

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.

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.

目的 建立黑巧克力中4种牛奶过敏原蛋白的超高效液相色谱-串联质谱检测方法.方法 从Uniprot数据库筛选牛奶主要过敏原蛋白的特异肽段,用Skyline工具模拟胰酶酶切,确定各肽段的多反应监测(multiple reaction monitoring,MRM)方法.50mmol/L碳酸氢铵(含2mol/L尿素)提取巧克力中 目标蛋白,14000 r/min、4℃离心除脂,胰酶酶解上清液,固相萃取柱净化,Peptide BEH C18(130 ?,2.1 mm×100 mm,1.7 μm)色谱柱分离,以乙腈和水[均含0.1％(V/V)甲酸]为流动相进行梯度分离,标准加入法定量.结果 4条特异肽段FFVAPFPEVFGK、NAVPITPTLNR、AVPYPQR和ALPMHIR可分别作为αS1-酪蛋白、αS2-酪蛋白、β-酪蛋白、β-乳球蛋白的定量肽段.其质量浓度与质谱响应峰面积呈现良好线性关系(R2＞0.99),方法的检出限和定量限分别为50～100和160～350 μg/kg,加标回收率为60.6％～116.7％,相对标准偏差均小于20％(n=6).结论 该方法灵敏、准确、高效,适于检测黑巧克力中4种牛奶过敏原蛋白.

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.

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.

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.

During the winemaking process, fining materials derived from milk and egg products are traditionally used to remove undesirable substances to reduce bitterness and astringency. The possible residues of allergens in treated wine may pose a potential risk for allergy patients. In this study, we developed a method for the simultaneous quantification of eight allergens (αS1-casein, αS2-casein, β-casein, κ-casein, β-lactoglobulin, lysozyme, ovalbumin and ovotransferrin) in red wine by ultrahigh-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS). The sample was extracted with polyvinylpolypyrrolidone (PVPP) solution, following trypsin digestion and peptide-level purification by solid-phase extraction (SPE). A strategy based on standard addition was used for the accurate quantification of the target allergens in wine products. The limits of detection (LODs) were shown to be 0.003-0.015 μg/mL for milk allergens and 0.1 μg/mL for egg allergens. This economical and reliable method would be appropriate for routine analysis and further allergen label management for red wine.

目的 建立丹磺酰氯衍生液相色谱-串联四极杆质谱法检测鸡肉、牛肉、牛奶中雌二醇的高灵敏分析方法.方法 酶解后的样品用HLB和氨基固相萃取柱富集净化,氮气吹干后,在60℃条件下丹磺酰氯衍生5 min;以乙腈-水(含0.1％乙酸)为流动相,采用多离子反应监测正离子模式进行检测.结果 鸡肉、牛肉、牛奶中雌二醇的检出限为0.6 ng/kg,定量限为2.0 ng/kg,3个添加水平的平均回收率在82.0％～88.9％之间,相对标准偏差均小于7.3％.我国6个城市牛肉、鸡肉和牛奶中17-β-雌二醇含量的中位数(平均值)分别为5.51 ng/kg(66.78 ng/kg)、4.60 ng/kg(16.94 ng/kg)和18.00 ng/kg(16.80 ng/kg),九十百分位数(P90)分别为248.91、39.33和39.60 ng/kg.结论 本方法适用动物肌肉和乳中超痕量雌二醇的含量检测.牛肉、鸡肉和牛奶中17-β-雌二醇的大样本监测数据可为科学的制定残留限量提供必要的数据基础.

Shellfish allergies constitute an important cause of food-induced anaphylactic reactions, which pose challenges to food safety and human health worldwide. In the present study, the specific IgE (sIgE) binding characteristics of different shrimp proteins of black tiger shrimp (Penaeus monodon) to the sera of eight shrimp-allergic patients from China were studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and nanoliquid chromatography time-of-flight mass spectrometry. According to the PLGS scores (>2000) and the sequence coverage (>40%), eight proteins with sIgE binding activity were identified, including myosin heavy chain type 1 (K4Q4N8), hemocyanin (G1AP69 and Q95V28), phosphopyruvate hydratase (O96656), arginine kinase (C7E3T4), tropomyosin (A1KYZ2), sarcoplasmic calcium binding protein (H7CHW2) and glyceraldehyde-3-phosphate dehydrogenase (A0A097BQP2). Among these eight proteins, phosphopyruvate hydratase was a prevalent IgE-binding protein among these Chinese patients with binding observed in 100% of sera. Moreover, 13 peptides were predicted as epitopes of phosphopyruvate hydratase. These new details help us to understand the crustacean IgE-binding proteins especially Penaeus monodon IgE-binding proteins, that would cause allergic reaction to Chinese patients. And our findings may provide essential information to improve allergy prevention and clinical treatment to shrimp allergy in China. PRACTICAL APPLICATION: This research may have diagnostic and therapeutic value for shrimp allergies in China.