3D-MPEA: A Graph Attention Model-Guided Computational Approach for Annotating Unknown Metabolites in Interactomics via Mass Spectrometry-Focused Multilayer Molecular Networking.

The spectral matching strategy of MS2 fragment spectrograms serves as a ubiquitous method for compound characterization within the matrix. Nevertheless, challenges arise due to the deficiency of distinctions in spectra across instruments caused by coelution peak-derived fragments and incompleteness of the current spectral reference database, leading to dilemma of multidimensional omics annotation. The graph attention model embedded with long short-term memory was proposed as an optimized approach involving integrating similar MS2 spectra into molecular networks according to the isotopic ion peak cluster spacing features to collapse diverse ion species and expand the spectral reference library, which efficiently evaluated the substance capture capacity to 123.1% than classic substance perception tactics. The versatility and utility of the established annotation procedure were showcased in a study on the stimulation of pork mediated by 2,2-bis(4-hydroxyphenyl)propane and enabled the global metabolite annotation from knowns to unknowns at metabolite-lipid-protein level. On the spectra for which in silico extended spectral library search provided a group truth, 83.5-117.1% accuracy surpassed 1.2-14.3% precision after manual validation. ß-Ala-His dipeptidase was first evidenced as the critical node related to the transformation of α-helical (36.57 to 35.74%) to random coil (41.53 to 42.36%) mediated by 2,2-bis(4-hydroxyphenyl)propane, ultimately triggering an augment of catalytic performance, inducing a series of oxidative stress, and further intervening in the availability of animal-derived substrates. The integration of ionic fragment feature networks and long short-term memory models allows the effective annotation of recurrent unknowns in organisms and the deciphering of unacquainted matter in multiomics.

High-confidence structural annotation of substances via multi-layer molecular network reveals the system-wide constituent alternations in milk interfered with diphenylolpropane.

The spectral database-based mass spectrometry (MS) matching strategy is versatile for structural annotating in ingredient fluctuation profiling mediated by external interferences. However, the systematic variability of MS pool attributable to aliasing peaks and inadequacy of present spectral database resulted in a substantial metabolic feature depletion. An amended procedure termed multiple-charges overlap peaks extraction algorithm (MCOP) was proposed involving identifying collision-trigged dissociation precursor ions through iteratively matching mass features of fragmentations to expand the spectral reference library. We showcased the versatility and utility of established strategy in an investigation centered on the stimulation of milk mediated by diphenylolpropane (BPA). MCOP enabled efficient unknown annotations at metabolite-lipid-protein level, which elevated the accuracy of substance annotation to 85.3% after manual validation. Arginase and α-amylase (|r| > 0.75, p < 0.05) were first identified as the crucial issues via graph neural network-based virtual screening in the abnormal metabolism of urea triggered by BPA, resulting in the accumulation of arginine (original: 1.7 µg kg-1 1.7 times) and maltodextrin (original: 6.9 µg kg-1 2.9 times) and thus, exciting the potential dietary risks. Conclusively, MCOP demonstrated generalisation and scalability and substantially advanced the discovery of unknown metabolites for complex matrix samples, thus deciphering dark matter in multi-omics.

High-throughput analysis of hazards in novel food based on the density functional theory and multimodal deep learning.

The emergence of cultured meat presents the potential for personalized food additive manufacturing, offering a solution to address future food resource scarcity. Processing raw materials and products in synthetic food products poses challenges in identifying hazards, impacting the entire industrial chain during the industry's further evolution. It is crucial to examine the correlation of biological information at different levels and to reveal the temporal dynamics jointly. Proposed active prevention method includes four aspects: (i) Investigating the molecular-level mechanism underlying the binding and dissociation of hazards with proteins represents a novel approach to mitigate matrix effect. (ii) Identifying distinct fragments is a pivotal advancement toward developing a novel screening strategy for hazards throughout the food chain. (iii) Designing an artificial intelligence model-based approach to acquire multi-dimensional histology data also holds significant potential for various applications. (iv) Integrating multimodal data is a practical approach to enhance evaluation and feedback control accuracy.

Long short-term memory based quasi-targeted lipidomics reveals propane-1,2-diol expediting the digestion of lipids via mediating the α-helices to a random curl or ß folding of lipase.

Scandal of detecting 1,2-propanediol (PD) in milk brought a crisis to the trust of consumers in dairy industry, and investigations focused effect of PD on digestive behavior of milk were still restricted. Long short-term memory amalgamated to quasi-targeted lipidomics was applied to monitor dynamics changes of lipids during digestion and the pseudo-first-order kinetic model elucidated that PD elevated the digestibility of lipid with the degradation rate (S-1) ranged from 4440.31 to 5665.59 and mediated the transition of α-helices (26.46% to 19.07% of pancreatic lipase and 29.89% to 23.37% of gastric lipase) covering active center in lipase to random curl (48.25% to 51.17% of pancreatic lipase and 41.58% to 44.57% of gastric lipase) and ß folding (9.14% to 4.67% of pancreatic lipase and 6.52% to 10.05% of gastric lipase), ultimately upregulating the lipase activity and further intervening lipid nutrients utilization in milk. This study provided a critical insight about the impact of PD contamination at trace concentrations on the nutritional value of milk fat during digestion.

Long Short-Term Memory-Based Multiomics Reveal Lactobacillus casei-Derived Postbiotics Inhibiting Lipids Digestion via Mediating the Upregulation of α-Helices in Lipase.

SCOPE: The antiobesity function of probiotics has been declared, while the application in high-risk patients and coding side effect has focused attention to postbiotics. This investigation profiles the mechanism of postbiotics affecting lipid digestion at molecular level, and establishes a momentous foundation for the clinical application of postbiotics in obesity suppression. METHODS AND RESULTS: An operational framework for butter digestion is constructed to collect the digests in the intestine at 0, 40, 80, and 120 min with various postbiotics supplement. A total of 227 lipids and 414 metabolites are detected by pseudo-targeted lipidomics integrated with the long short-term memory-based metabolomics, and the triacylglycerol (TG, from 134.1 to 184.7 mg kg-1 ) and diacylglycerol (DG, from 4.2 to 8.4 mg kg-1 ) are identified as significantly different lipids with or without postbiotics supplement. A total of eight substances related to the inhibition of gastric lipase and pancreatic lipase are screened through the molecular simulation computation in silicon and enzymatic reaction kinetics, and thus curtailing the bioaccessibility of lipids. CONCLUSIONS: Lactobacillus casei JCM1134-derived postbiotics propel the structure of lipase to aggregate by increasing the α-helix, and thus hampering the digestion of triglycerides through noncompetitive inhibition.

Lactobacillus casei-derived postbiotics inhibited digestion of triglycerides, glycerol phospholipids and sterol lipids via allosteric regulation of BSSL, PTL and PLA2 to prevent obesity: perspectives on deep learning integrated multi-omics.

The anti-obesity potential of probiotics has been widely reported, however their utilization in high-risk patients and potential adverse reactions have led researchers to focus their attention on postbiotics. Herein, pseudo-targeted lipidomics linked with deep learning-based metabolomics was utilized to dynamically characterize the postbiotic potential of heat-inactivated Lactobacillus casei JCM1134 supplementation after a high-fat diet in treating obesity. MG (ranged from 423.0 ± 1.4 mg L-1 to 331.45 ± 2.3 mg L-1), LPC (ranged from 13.1 ± 0.08 mg L-1 to 10.2 ± 0.1 mg L-1) and Cho (ranged from 9.0 ± 0.3 mg L-1 to 5.7 ± 0.2 mg L-1) in intestinal digestive products were significantly decreased, indicating that the digestion of lipid was inhibited. 8-C-glucosylorobol, from Lactobacillus casei, was confirmed from quantitative results and molecular simulation calculations to inhibit the transformation of TG, DG, and ChE through weakening hydrogen bonds between enzymes and substrates and reducing the binding energy. Pristimerin and 2,4-quinolinediol can effectively reduce the hydrogen bonding force between PC and phospholipase A2, which were related to the obstruction of phosphatidylcholine digestion. This research deepened the understanding of the mechanism underlying the inactivated probiotics affecting lipid digestion, establishing the critical groundwork for clinical application of probiotics in inhibiting obesity.

Lactobacillus casei-Derived Postbiotics Elevate the Bioaccessibility of Proteins via Allosteric Regulation of Pepsin and Trypsin and Introduction of Endopeptidases.

The potential of probiotics to benefit digestion has been widely reported, while its utilization in high-risk patients and potential adverse reactions have focused interest on postbiotics. A variable data-independent acquisition (vDIA)-based spatial-omics strategy integrated with unsupervised variational autoencoders was applied to profile the functional mechanism underlying the action of Lactobacillus casei-derived postbiotic supplementation in goat milk digestion in an infant digestive system, from a metabolomics-peptidomics-proteomics perspective. Amide and olefin derivatives were proved to elevate the activities of pepsin and trypsin through hydrogen bonding and hydrophobic forces based on allosteric effects, and recognition of nine endopeptidases and their cleavage to serine, proline, and aspartate were introduced by postbiotics, thereby promoting the generation of hydrophilic peptides and elevating the bioaccessibility of goat milk protein. The peptides originating from αs1-casein, ß-casein, ß-lactoglobulin, Ig-like domain-containing protein, κ-casein, and serum amyloid A protein, with multiple bioactivities including angiotensin I-converting enzyme (ACE)-inhibitory, osteoanabolic, dipeptidyl peptidase IV (DPP-IV) inhibitory, antimicrobial, bradykinin-potentiating, antioxidant, and anti-inflammatory activities, were significantly increased in the postbiotic supplementation group, which was also considered to potentially prevent necrotizing enterocolitis through inhibiting the multiplication of pathogenic bacteria and blocking signal transducer and activator of transcription 1 and nuclear factor kappa-light-chain-enhancer of activated B cells inflammatory pathways. This research deepened the understanding of the mechanism underlying the postbiotics affecting goat milk digestion, which established a critical groundwork for the clinical application of postbiotics in infant complementary foods.

Ambient 1,2-propanediol exposure accelerates the degradation of lipids and amino acids in milk via allosteric effects and affects the utilization of nutrients containing amide bond.

The scandal of detecting 1, 2-propanediol (PL) in milk brought a crisis to the trust of consumers in the dairy industry, and the potential toxicity of PL has aroused the public concern about dietary exposure. A total of 200 pasteurized milk samples were collected from 15 regions, and the quantity of PL ranged between 0 and 0.31 g kg-1. Pseudo-targeted quantitative metabolomics integrated with proteomics demonstrated that PL enhanced the reduction of κ-casein, ß-casein, and 107 substances (41 amines and 66 amides) containing amide bonds. Pathway enrichment and topological analysis indicated that PL induced the metabolism of lipids, amino acids, oligosaccharide nucleotides, and alkaloids by accelerating the rate of nucleophilic reaction, and acetylcholinesterase, sarcosine oxidase, and prolyl 4-hydroxylase were determined as the vital enzymes related to the degradation of above nutrients. The results of molecular simulation calculation illustrated that the number of hydrogen bonds between acetylcholinesterase, sarcosine oxidase, and substrate increased to 2 and 3, respectively, while the position of hydrogen bonds between prolyl 4-hydroxylase and proline was shifted, indicating the change of conformation and the enhancement of hydrogen bond force were essential factors for the up-regulation of enzyme activity. This study first revealed the mechanism of deposition and transformation of PL in milk, which contributed to the knowledge of the quality control of milk and provided vital indicators to evaluate the adverse risks of PL in dairy products.

Extracellular proteolytic enzyme-mediated amino exposure and ß-oxidation drive the raspberry aroma and creamy flavor formation.

The fermentation-driving ability of Daqu has been widely reported, while the potential influence of substances in Daqu on Baijiu flavor formation has attracted increasing interest. Pseudo-targeted metabolomics integrated proteomics combined with sensory evaluation strategy was applied to investigate the correlation between flavor characteristics and metabolic profiling of Daqu, and the mechanism of flavor formation was also elucidated. The 4-hydroxy-2,5-dimethylfuran-3-one (3.5 mg kg-1) and 2,3-dihydro-1 h-inden-5-ol (894.3 µg kg-1) were identified as the unique substances in qingcha qu, which were vital for raspberry flavor formation and associated with the up-regulation of amino acid metabolism. The dec-9-enoic acid (37.4 mg kg-1) was screened out as the substance related to the formation of cream flavor in hongxin qu produced through the shortening of fatty acid carbon chains and unsaturated modification of long chain fatty and acceleration of carbon metabolism in hongxin qu mediated by filamentous Aspergillus spp. was related to the smoky aroma enhancement.

Goat milk-derived short chain peptides: Peptide LPYV as species-specific characteristic and their versatility bioactivities by MOF@Fe3O4@GO mesoporous magnetic-based peptidomics.

Goat milk as an ideal substitute for human milk has not been sufficiently explored. An in-situ synthesized MOF@Fe3O4@GO was demonstrated as a magnetic mesoporous adsorbent for efficiently enriching short chain peptides (SCPs) in milk compared with the routine solid phase extraction approach with graphite carbon black or C18 as the packing material in terms of the number of enriched SCPs and data stability. A total of 61 and 126 SCPs were identified and quantified in bovine milk (0.09-89.34 µg L-1) and goat milk (10.5-1267.06 µg L-1), respectively, and peptide LPYV can be used as a potential marker for adulteration of goat milk. Relative high expression of chymotrypsin and pepsin by EnzymePredictor analysis could partially elaborate the reason of the abundance of SCPs in goat milk. Compared with bovine milk, further bioinformatics analysis indicated that goat milk could own higher nutritional value because of relative higher concentrations (>1 mg/L) of SCPs (LLV, FL, LVYP) with confirmed bioactivities including angiotensin-converting enzyme (ACE) inhibitor, antioxidant, dipeptidylpeptidase (DPP) III and DPP IV inhibitor, etc. Overall, this study opened a novel avenue for understanding versatility benefit of dairy products from a perspective of SCPs by using a developed MOF@Fe3O4@GO mesoporous magnetic-based peptidomics.

UHPLC-Q-Orbitrap-Based Integrated Lipidomics and Proteomics Reveal Propane-1,2-diol Exposure Accelerating Degradation of Lipids via the Allosteric Effect and Reducing the Nutritional Value of Milk.

The scandal of detecting the flavoring solvent propane-1,2-diol (PD) in milk has brought a crisis to the trust of consumers in the dairy industry, while its deposition and transformation are still indistinct. Pseudo-targeted lipidomics revealed that PD accelerated the degradation of glycerolipid (33,638.3 ± 28.9 to 104,54.2 ± 28.4 mg kg-1), phosphoglyceride (467.4 ± 8.2 to 56.6 ± 4.2 mg kg-1), and sphingolipids (11.4 ± 0.3 to 0.7 ± 0.2 mg kg-1), which extremely decreased the milk quality. Recoveries and relative standard deviations (RSDs) of the established method were 85.0-109.9 and 0.1-14.9%, respectively, indicating that the approach was credible. Protein-lipid interactions demonstrated that 10 proteins originating from fat globules were upregulated significantly and the activities of 7 enzymes related to lipid degradation were improved. Diacylglycerol cholinephosphotransferase was the only enzyme with decreased activity, and the molecular docking results indicated that PD adjusted its activity through regulating the conformation of the active center and weakening the hydrogen bond force between the enzyme and substrate. This study firstly revealed the mechanism of deposition and transformation of PD in milk, which contributed to the knowledge on the milk quality control and provided key indicators to evaluate the adverse risks of PD in dairy products.

Pseudo-targeted metabolomics analysis of the therapeutic effect of phenolics-rich extract from Se-enriched green tea (Camellia sinensis) on LPS-stimulated murine macrophage (RAW264.7).

Selenium (Se)-enriched green tea (Se-Te) has been recognized as a possible source of Se supplements, while the effect of Se enrichment on function of polyphenols in green tea is still unclear. In this study, a pseudo-targeted metabolomics strategy was carried out to reveal the regulatory mechanism of polyphenols extracted from Se-Te and regular green tea (Re-Te) on inflammatory response at cellular level. A novel analysis strategy using UHPLC/ESI Q-Orbitrap combined with MS-IOP was applied to profile the dynamic changes of metabolites in LPS-stimulated RAW264.7 macrophages during polyphenols incubation. A total of 128 characteristic variables (VIP > 1, p < 0.05) were screened in Se-Te group and the results of bioinformatics analysis and quantitative research indicated that in addition to the 6 conventional immune protective pathways involved in tea polyphenols, Se-enriched polyphenols were also participated in 3 unique antioxidant enzyme activation pathways, including phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism and pantothenate and CoA biosynthesis. The result of weight calculation based on topological analysis indicated that the promoting synthesis of antioxidant enzymes was the main mechanism of Se-Te polyphenols to inhibit inflammation. However, compared with Re-Te group, the intracellular B vitamin pathway in Se-Te group was disturbed, which is related to the fact that Se supplementation can promote the synthesis of selenoprotein and catalyze the reduction of thioredoxin by NADPH, thus blocking the signaling pathways of B vitamins. This study comprehensively explored the immune protective mechanism of polyphenols extracted from Se-Te and Re-Te under natural growth conditions, which could give a better understanding of the potential nutritional value of Se-Te as a widely used Se supplement.

Effects of Short-Chain Peptides on the Flavor Profile of Baijiu by the Density Functional Theory: Peptidomics, Sensomics, Flavor Reconstitution, and Sensory Evaluation.

The effect of peptides on the flavor profile of Baijiu is unclear as a result of their trace concentrations in the complex matrix, and therefore, the study involving the interaction mechanism between peptides and flavor compounds is limited. In this study, short-chain peptides (amino acid number between 2 and 4, SCPs) associated with the Feng-flavor Baijiu (FFB) were comprehensively analyzed by a dedicated workflow using ultra-high-performance liquid chromatography Q Orbitrap high-resolution mass spectrometry, flavor reconstitution experiments, sensory analysis, and density functional theory (DFT) analysis. The concentrations of 96 SCPs intimately related with six different grades of honey aroma intensity in FFB were quantified (0.12-155.01 µg L-1) after multivariable analysis, Spearman's correlation analysis (ρ ≥ 0.7), and confirmation with synthetic standards, and 32 dominant odorants with an odor activity value of ≥1 in FFB with the highest intensity of honey aroma were quantified by gas chromatography-mass spectrometry and gas chromatography-flame ionization detection analyses. The results of flavor reconstitution experiments and sensory analysis indicated that the SCPs can obviously influence the honey aroma with amplifying the fruity, sweet, and flora flavor odor characters (p < 0.05) while significantly reducing the acidic character (p < 0.001), which could be attributed to the most stable complex structure between SCPs and odor-active compounds calculated by DFT being butanoic acid, followed by ß-damascenone, 3-methylbutanal, and ethyl hexanoate, and the multiple sites as a hydrogen bond donor or acceptor in SCPs can form a stable ternary structure with water and ethanol inside the peptide chain or carboxyl terminal of SCPs, consequently improving the stability of the Baijiu system. The results highlighted the important role of SCPs on the volatiles in Baijiu and laid the foundation for further facilitating the sensory quality of Baijiu products.

Discovery of Se-containing flavone in Se-enriched green tea and the potential application value in the immune regulation.

Selenium (Se)-enriched green tea has been recognized as a possible source of supplemental Se, while the structural and physiological activities of Se-containing flavone are still unclear. In this study, a Se-containing flavone was isolated from Se-enriched green tea by high-speed counter-current chromatography (HSCCC) and characterized through UHPLC-Q-Orbitrap, FT-IR and NMR. Results proved that HSeO3- can be combined with the alcohol hydroxyl of 2-phenylchromone in flavone and the content of Se-containing flavone in tea was 15690.4 µg L-1. Additionally, Se-containing flavone can effectively inhibit the production of nitric oxide (NO), and downregulate expression of TNF-α and IL-6. Compared with regular flavone extracted from green tea (43.24 pg mL-1), release of IL-10 was higher in Se-containing flavone group (53.37 pg mL-1), indicating that Se-containing flavone played an important role in the process of severe inflammatory injury. The results indicated that Se-containing flavone was an attractive natural ingredient for developing novel functional foods.

Novel top-down high-resolution mass spectrometry-based metabolomics and lipidomics reveal molecular change mechanism in A2 milk after CSN2 gene mutation.

The effects of milk containing only native A2 beta casein (A2 milk) on amelioration of gut tolerance are widely studied by clinical in vivo trials. However, the molecular change mechanism after CSN2 gene mutation and other potential health benefits in A2 milk remain unknown. The glycerophospholipid metabolism, sphingolipid metabolism, glycerolipid metabolism, phosphatidylinositol signaling system and glycosylphosphatidylinositol (GPI)-anchor biosynthesis were considered as significant metabolic pathways intimately related with the mutation of CSN2 gene by integrating metabolomics and lipidomics and the corresponding molecular mechanism was summarized. The contents of lactose, triacylglycerol and most polar lipids significantly decreased after CSN2 gene mutation by utilizing UHPLC-Q-Orbitrap HRMS (LOQs, 0.0004-0.0215 µg/mL) combined with multivariate analysis, furthermore, the essential fatty acids α-linolenic acid (1015.22 to 543.14 µg/mL) and arachidonic acid were significantly down regulated although eicosapentaenoic acid was relatively upregulated, which provided a novel perspective to understand the nutritional value and health benefits of A2 milk.

Novel insight into the transformation of peptides and potential benefits in brown fermented goat milk by mesoporous magnetic dispersive solid phase extraction-based peptidomics.

Brown fermented goat milk as an excellent source of bioactive peptides has only been partially elucidated. Meticulously synthesized MOF@MG as magnetic sorbent for enriching endogenous peptides owned higher reproducibility and uniform distribution of peptides PI compared with ultrafiltration. Combined with UHPLC-Q-Orbitrap, fermentation for 12 h in brown goat milk with the highest overall acceptable degree through sensory evaluation was utilized to explore the transformation of peptides and health benefits, with trypsin or plasmin hydrolyzing proteins and aminopeptidase or carboxypeptidase hydrolyzing peptides to small peptides or amino acids. A total of 1317 peptides were identified by database matching (1259) and de novo sequencing (58), among 18 peptides could originate from gene-independent enzymatic formation and top 25 characteristic peptides were quantified with concentration ranging from 0.12 to 6.40 mg L-1. Bioinformatic analysis results indicated that brown fermented goat milk possesses higher health benefits because of more than 50 peptides with potential bioactivity.

Molecular mechanism of high pressure shear grinding on Feng-flavour Chinese Baijiu ageing.

Ultrahigh-pressure homogenization is a powerful technique for accelerating Baijiu ageing. In this study, the molecular mechanism of artificial ageing was thoroughly investigated utilizing foodomics combined with a mass spectrum information optimization program. Various characterizations of compounds in the ageing process were performed through ultra high performance liquid chromatography quadrupole-orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap) based colloid theoretics. High-pressure shear grinding provides a high-pressure and high-temperature environment for Baijiu, accelerating the formation of flavour compounds and the decomposition of odour and harmful substances, thus improving the sensory characteristics of Feng-flavour Baijiu and shortening the ageing time. A total of 2069 substances were identified according to fragment spectra and Bayesian network model-based peak distribution. Molecules with VIP > 1, including 14 esters, 10 acids, 4 ketones, 3 aldehydes, 2 alcohols and 2 phenols, were quantified as potential markers of chemometric analysis. A novel prediction model for Baijiu ageing degree was furnished, and ageing years of Feng-flavour Chinese Baijiu were accurately predicted by factor analysis with KMO 0.784 and a polynomial regression model with r 0.94. The results demonstrated that in 400 bar, fresh distilled Baijiu could reach a natural ageing effect of 6.43 years, which was consistent with the sensory evaluation.

Molecular mechanism of Mare Nectaris and magnetic field on the formation of ethyl carbamate during 19 years aging of Feng-flavor Baijiu.

Ethyl carbamate (EC) is carcinogen occurring naturally in fermented foods, while the EC formation pattern in Feng-flavor Baijiu during Mare Nectaris storage and magnetic field treatment remains controversial. In this work, variation of EC in Mare Nectaris and magnetic field were investigated for the first time through ultra high performance liquid chromatography quadrupole-orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap). Quantification results revealed that EC decreased significantly in the stage of 3-9 years and kept at 12.4 µg L-1 after 10 years of aging. Arginine succinate synthase (ASS) and urease were deemed as vital factors for EC decomposition. Degradation effetc of EC in 250 mT is simillar to that of EC in Baijiu stored in Mare Nectaris for 8 years. This is due to that aging process was accelerated by magnetic field and the content of total acid in Baijiu was increased, creating a favorable environment for decomposition of EC and urea.

Characterisation of key odorants causing honey aroma in Feng-flavour Baijiu during the 17-year ageing process by multivariate analysis combined with foodomics.

Honey aroma is a typical sensory characteristic of Feng-flavour Baijiu, which originates from a unique manufacturing process, the formation mechanism of which is unclear. Multivariate analysis combined with foodomics assisted by sensory evaluation was performed to investigate the molecular mechanism of honey aroma formation in Feng-flavour Baijiu during the 17-year ageing process. A total of 1995 compounds was identified, and 47 variables were screened as significant substances according to variable importance in projection and Spearman's rank correlation coefficient (|ρ| > 0.7), which corroborated that the long-term interaction between Baijiu and storage containers was the dominant origin of honey aroma. Recombination and omission experiments further validated the important contributions of significant substances, including acids, alcohols, aldehydes and ketones. A typical honey aroma dominated by fruity, floral, sweet and nutty notes was successfully simulated, and nutty notes could be enhanced by amides, whereas amines presented masking effects on fruity and floral aromas.

Physicochemical and molecular transformation of novel functional peptides from Baijiu.

A novel strategy for screening and identifying peptides present in Baijiu was developed based on magnetic solid-phase extraction with magnetic S-doped graphene (M-G-S) as adsorbent combined with ultrahigh-performance liquid chromatography with high resolution tandem mass spectrometry. In total, 28 peptides consisting of amino acids from 3 to 9 were preliminarily identified, and significantly higher in the number than that of direct concentration and SPE with C18 as the adsorbent. Six peptides were confirmed with their corresponding synthetic reference standards by comparing their retention time, high resolution MS/MS spectra, and NMR spectroscopic studies. Parallel reaction monitoring integrated with the internal standard method was utilized to quantify identified peptides with concentrations ranging from 1.14 to 10.25 ng mL-1, and prediction results of bioactivity comprising antioxidation or ACE inhibitors were obtained. These discoveries were conducive to understanding the versatility benefit of Baijiu and paved the way to study the interaction between peptides and volatile substances.