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Abstract Background Metabolomics is an emerging field that quantifies numerous metabolites systematically aiming to determine the metabolites corresponding to each biological phenotype and then provide an analysis of the mechanisms involved. Bile acids (as an organic metabolites) are synthesized in the liver from cholesterol and could be used as indicator of hepatobiliary impairment. However, the role of these bile acids in the pathogenesis of cholangiocarcinoma (CCA) and hepatocellular carcinoma (HCC) is still unclear. Therefore, the current study aimed to use serum bile acid profiles potential diagnostic biomarkers for early detection of cholangiocarcinoma and differentiating it from hepatocellular carcinoma. Subjects and methods Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC/MS/ MS) analytical method was used for the measurement of bile acids in the serum of patients with hepatocellular carcinoma (n = 35), cholangiocarcinoma (n = 35), and control group (n = 35) to determine role as markers for differentiation between hepatocellular carcinoma and cholangiocarcinoma. Results This study revealed that there was a significant increase in all 14 bile acids in both HCC and CCA compared to control. Also, there was significant increase in LCA, TCA, GDCA, and GCA in cholangiocarcinoma (CCA) compared to HCC with AUC 0.775, 0.825, 0.797, and 0.831 respectively with highest sensitivity and specificity for GCA (82% and 74%, respectively) for differentiation between the two types of cancers. Conclusion Determination of the serum bile acids pattern using UPLC/MS/MS may help to differentiate between hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA) especially GCA which may be a good biomarker for differentiation between two types of liver cancers.

Abstract Background Bile acids are vital regulators of liver metabolism, and their dysregulation is closely linked with the progression of nonalcoholic fatty liver disease (NAFLD). Profiling these bile acids may provide valuable diagnostic and prognostic markers for these conditions. This study aimed to evaluate bile acid profiles in NAFLD patients and assess their potential as biomarkers for diagnosing and predicting disease progression. Serum levels of 14 bile acids were measured in 25 normal healthy controls (NHC), 35patients with metabolic dysfunction–associated steatotic liver disease (MASLD), and 40 patients with NASH, categorized by the NAFLD Activity Score (NAS). Quantification was performed using high-performance liquid chromatography-tandem mass spectrometry (HPLC–MS/MS). Results Primary unconjugated bile acids, CA and CDCA, along with conjugated acids GCA, GCDCA, TCA, and TCDCA, were significantly elevated in both MASLD and NASH compared to NHC (all p

Abstract Background Effects of the renin‐angiotensin‐aldosterone system (RAAS) inhibitors enalapril and telmisartan on circulating RAAS in dogs with proteinuric chronic kidney disease (pCKD) are undescribed. Objectives To characterize the RAAS in untreated dogs with pCKD compared to healthy, life‐stage‐ and sex‐matched controls, and in dogs with pCKD after 30 days of treatment with enalapril or telmisartan. Animals Dogs with pCKD (n = 36) and healthy controls (n = 20). Methods Retrospective study of banked samples and previously collected data. Day 0 serum equilibrium concentrations of angiotensin I, II, III, IV, 1‐5, and 1‐7, and aldosterone, and urinary aldosterone‐to‐creatinine ratio (UACR) from pCKD dogs were compared to values on day 30 of treatment with enalapril (0.5 mg/kg PO q12) or telmisartan (1 mg/kg PO q24h) and to those of healthy dogs. Data were analyzed using linear mixed models. Results Compared with healthy dogs, pCKD dogs had significantly higher Ang I, III, 1‐5, and 1‐7 concentrations, and UACR. Relative to pretreatment values, day 30 Ang II concentrations were significantly increased and decreased in telmisartan‐ and enalapril‐treated pCKD dogs, respectively (both P

Salvia Miltiorrhiza, extensively distributed and commonly employed as a traditional Chinese medicinal herb, has garnered significant attention. In this study, the Salvia Miltiorrhiza extract (SME) was prepared by one-step water extraction method, and was firstly used as a novel corrosion inhibitor for Q235 steel in 1 mol/L HCl solution. As identified via liquid chromatography-mass spectrometry, the findings reveal that Salvianic acid A, tanshinone II A, danshenxinkun D, dihydrotanshinone, and methylene tanshinquinone are the primary constituents of SME. The optimum corrosion inhibition efficiency reached 92.8 % at 200 mg/L and maintained at 90.4 % after 72 h. Based on in-situ scanning vibration electrode technology (SVET) monitoring, the adsorption of inhibitor molecules on metal surface greatly retarded the propagation of localized corrosion. AFM examination of the corroded surface reveals that the samples supplemented with SME exhibit a smoother surface compared to the blank group. The force curve graph for the SME-added group demonstrates a more evenly distributed point array and an elevated average adhesion force, indicating that the addition of SME improves the corrosion resistance of the metal surface. XPS characterization illustrated that SME interacted with iron ions to form insoluble precipitate. This work investigated the application of SME as an eco-friendly corrosion inhibitor for carbon steel in acidic medium, providing a new approach for the high value-added utilization of Salvia Milliorrhiza.

Piperazines are a class of new psychoactive substances with hallucinogenic effects that affect the central nervous system by affecting the level of monoamine neurotransmitters. Abuse of piperazines will produce stimulating and hallucinogenic effects, accompanied by headache, dizziness, anxiety, insomnia, vomiting, chest pain, tachycardia, hypertension and other adverse reactions, and may even cause cardiovascular diseases and multiple organ failure and lead to death, seriously affecting human physical and mental health and public safety. The abuse of new psychoactive substance piperazines has attracted extensive attention from the international community. The study of its pharmacological toxicology and analytical methods has become a research hotspot in the field of forensic medicine. This paper reviews the in vivo processes, sample treatment and analytical methods of existing piperazines, in order to provide reference for forensic identification.

In order to investigate the formation mechanism of the quality of baked green tea, we used metabolomics based on ultra-high performance liquid chromatography-quadrupole-Exactive mass spectrometry (UPLC-Q-Exactive/MS) to analyze the differences in the chemical compositions of green tea samples at different stages of thermal processing such as fixation, drying and roasting. A total of 125 compounds were identified, including 10 flavanols, 14 dimeric catechins, 19 flavanol-O-glycosides, 5 flavone-C-glycosides, 8 N-ethyl-2-pyrrolidinone-substituted flavan-3-ols (EPSF), 16 amino acids, 13 phenolic acids, 4 organic acids, 11 alkaloids, 13 lipids, 4 aroma glycosides and 8 other compounds. The results of partial least squares-discriminant analysis (PLS-DA) and heat map analysis showed that the chemical components obviously changed during the thermal processing of green tea, and 114 significantly differential compounds were selected (P < 0.05). In the three stages of thermal processing, the contents of most catechins and dimeric catechins decreased, and the contents of EPSFs, flavone- C-glycosides, and lipid compounds significantly increased. The contents of aroma glycosides (phenylethyl primeveroside, linalool primeveroside, and linalool oxide primeveroside) increased by 439% to 2 497% during the fixation process. The content of N-(1-deoxy-D-fructosyl)theanine increased by 820% to 1 290% during the drying process. The contents of flavanol-O-glycosides (myricetin 3-galactoside, kaempferol-3-galactoside, kaempferol 3-arabinoside, myricetin 3-glucoside and kaempferol-3-glucoside) decreased significantly during roasting. This study can provide a theoretical reference for future improvement of green tea quality.

The chemoselective probe technique combines the benefits of chemical derivatization and solid-phase extraction, allowing for simultaneous enhancement of analyte ionization efficiency, reduction of matrix interference, and improvement in chromatographic separation. This innovative technique holds great promise for applications in metabolomics research, metabolic tracking, and natural product isolation and purification. This review provides a comprehensive overview of the advancements in chemoselective probes from 2007 to 2024, focusing on the progress in probe design, particularly cleavable linkers. Additionally, it discusses the application of chemoselective probes in LC-MS analysis of small-molecular-weight compounds.

Pancreatic hormones are produced in the islets of Langerhans in the pancreas and are secreted to regulate blood sugar levels. Key pancreatic hormones are insulin, glucagon, and somatostatin. The measurement of these peptides/small proteins is essential in the study of the endocrine system, regulating diseases such as diabetes mellitus, but also in doping control and forensics. In this review, we focus on approaches for measuring pancreatic hormones using liquid chromatography-mass spectrometry (LC–MS). A rich variety of LC–MS approaches are used, including variations in LC column dimensions and chemistry, factors that are crucial regarding sensitivity, selectivity, robustness, and speed. In addition, several sample preparation methods are used (protein precipitation, immunopurification, etc.), for the analysis of matrices such as blood, urine, and cell culture medium. There are also several variants for mass spectrometric analysis, both targeted and non-targeted, using both lower and high-resolution instruments. LC–MS allows for sensitive, multi-hormone measurements, confidently distinguishing between hormones and their analogs, metabolites, and degradation products. The speed of analysis can be pushed down to a few minutes, with methods being validated and clinically applied. The review focuses on papers that have been published between 2019 and 2024.

Background Polycystic ovary syndrome (PCOS) is the most common metabolic disorder and reproductive endocrine disease, posing an elevated risk to women of reproductive age. Although metabolism differences in serum, amniotic fluid and urine have been documented in PCOS, there remains a paucity of evidence for vaginal fluid. This study aimed to identify the metabolic characteristics and potential biomarkers of PCOS in Chinese women of reproductive age. Methods We involved ten newly diagnosed PCOS women who attended gynecology at Zhongda Hospital and matched them with ten healthy controls who conducted health check-up programs at Gulou Maternal and Child Health Center in Nanjing, China from January 1st, 2019 to July 31st, 2020. Non-targeted metabolomics based on ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was applied to differentially screen vaginal metabolites between PCOS group and healthy controls. Principal component analysis (PCA), orthogonal partial least-squares discriminant analysis (OPLS-DA) and enrichment analysis were used to observe differences, search for potential biomarkers and enrich related pathways. Results Among the 20 participants, a total of 195 different metabolites were detected between PCOS group and healthy control group. PCOS and control groups were effectively separated by vaginal fluid. Lipids and lipid-like molecules constituted the majority of differential metabolites. Notably, dopamine exhibited an increased trend in PCOS group and emerged as the most significant differential metabolite, suggesting its potential as a biomarker for identifying PCOS. The application of UHPLC-MS/MS based vaginal metabolomics methods showed significant differences between PCOS and non-PCOS healthy control groups, especially linoleic acid metabolism disorder. Most differential metabolites were enriched in pathways associated with linoleic acid metabolism, phenylalanine metabolism, tyrosine metabolism, nicotinate and nicotinamide metabolism or arachidonic acid metabolism. Conclusions In this pilot investigation, significant metabolomics differences could be obtained between PCOS and healthy control groups. For PCOS women of reproductive age, vaginal metabolism is a more economical, convenient and harmless alternative to provide careful personalized health diagnosis and potential targets for therapeutic intervention.

Chlorella vulgaris is renowned for its rich nutritional composition and has been associated with various health benefits. This study aimed to look into the effect of CV on liver metabolomes and metabolic pathways in aged Sprague Dawley (SD) rats. Three and 21-month-old male SD rats were divided into control and treated groups. Results showed that carbohydrates such as maltose/lactose and mannose/fructose were downregulated in old rats. These metabolites were downregulated in young rats supplemented with CV. Interestingly, taurine was upregulated in old rats, while the trend was reversed in old rats supplemented with CV at 300 mg/kg/d. The differentially expressed metabolites in different age groups and CV supplementation were mapped to starch and sucrose metabolism, galactose metabolism, and taurine and hypotaurine metabolism. Further research is needed to understand the long-term effects of CV supplementation on ageing and determine optimal dosage levels, especially regarding its potential to mitigate age-related diseases.

Age related cataracts (ARC) represent the main reason for blindness globally. The lens epithelial cells (LECs) participate not only in the metabolism of many substances in the lens but also in maintaining lens transparency. This study used lipidomics to investigate the metabolic differences in LECs of ARC patients with different severity, aiming at identifying potential metabolic biomarkers of ARC. Patients diagnosed with ARC and underwent cataract surgery at Shanghai Tongren Hospital were selected to participate in this study, which were classified as mild ARC group and severe ARC group. During their cataract surgery, anterior lens capsules(LCs) containing LECs were obtained. The lipidomics of LECs were analyzed using the liquid chromatography‑mass spectrometry (LC-MS). Potential pathways of lipids were searched for using databases such as the Kyoto Encyclopedia of Genes and Genomes (KEGG) and MetaboAnalyst platform. In LEC lipids, 26 lipids have been identified as potential biomarkers between mild ARC and severe ARC, with AUC values of 0.67–0.94. The pathway analysis results revealed that the Glycerophospholipid (GPL) metabolism was significantly influenced, indicating that these metabolic markers contribute significantly to regulating this pathway. The LEC metabolic spectrum demonstrates a proficient ability to differentiate between patients with varying levels of cataracts. Herein, we have successfully identified potential metabolic biomarkers and pathways that have proven to be valuable in enhancing our understanding of ARC pathogenesis. The finding has translational value for developing new cataract treatment methods in the future.

Abstract Background The transcriptome and metabolome dissection of the skeletal muscle of high- and low- growing individuals from a crossbred population of the indigenous Chongming white goat and the Boer goat were performed to discover the potential functional differentially expressed genes (DEGs) and differential expression metabolites (DEMs). Results A total of 2812 DEGs were detected in 6 groups at three time stages (3,6,12 Month) in skeletal muscle using the RNA-seq method. A DEGs set containing seven muscle function related genes (TNNT1, TNNC1, TNNI1, MYBPC2, MYL2, MHY7, and CSRP3) was discovered, and their expression tended to increase as goat muscle development progressed. Seven DEGs (TNNT1, FABP3, TPM3, DES, PPP1R27, RCAN1, LMOD2) in the skeletal muscle of goats in the fast-growing and slow-growing groups was verified their expression difference by reverse transcription-quantitative polymerase chain reaction. Further, through the Liquid chromatography-mass spectrometry (LC-MS) approach, a total of 183 DEMs in various groups of the muscle samples and these DEMs such as Queuine and Keto-PGF1α, which demonstrated different abundance between the goat fast-growing group and slow-growing group. Through weighted correlation network analysis (WGCNA), the study correlated the DEGs with the DEMs and identified 4 DEGs modules associated with 18 metabolites. Conclusion This study benefits to dissection candidate genes and regulatory networks related to goat meat production performance, and the joint analysis of transcriptomic and metabolomic data provided insights into the study of goat muscle development.

Objective: The objective of this study was to decipher chemical interactions between Danshen and Danggui using liquid chromatography–mass spectrometry (LC-MS) and explore the mechanisms of Danshen–Danggui against stroke using network pharmacology and molecular docking. Materials and Methods: First, the chemical compounds of Danshen–Danggui were profiled using ultra-high-performance liquid chromatography (HPLC)-quadrupole time-of-flight MS. Accurately characterized compounds in various proportions of Danshen–Danggui were quantified using HPLC combined with triple quadrupole electrospray tandem MS. Network pharmacology was used to uncover the essential mechanisms of action of Danshen–Danggui against stroke. Discovery Studio Software was used for the molecular docking verification of key active chemicals and stroke-related targets. Results: A total of 53 compounds were characterized, and 22 accurately identified constituents (10 phenolic acids, 8 phthalides, and 4 tanshinones) were quantified in 15 proportions of Danshen–Danggui. The quantification results showed that Danggui significantly increased the dissolution of most phenolic acids (compounds from Danshen), whereas Danshen promoted the dissolution of most phthalides (compounds from Danggui). Overall, the combination of Danshen and Danggui at a 1:1 ratio resulted in the maximum total dissolution rate. Further network pharmacology and molecular docking results indicated that Danshen–Danggui exerted anti-stroke effects mainly by regulating inflammation-related (tumor necrosis factor, hypoxia-inducible factor, and toll-like receptor) signaling pathways, which ranked among the top three pathways based on Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Conclusion: The chemical compounds in Danshen–Danggui could interact with each other to increase the dissolution of the most active compounds, which could provide a solid basis for uncovering the compatibility mechanisms of Danshen–Danggui and Danshen–Danggui-based formulae.

In order to determine compounds present in the aqueous extract of Cinnamomum kanehirae Hay (CWE) that promote the asexual sporulation of Antrodia cinnamomea in submerged fermentation. First, CWE was isolated by alcohol precipitation and fractional extraction with different organic solvents. The influence of the obtained fractions on the asexual sporulation of A. cinnamomea was investigated. It was showed that the chloroform (at 30 μg/mL) and ethyl acetate (at 50 μg/mL) extracts of the supernatant after ethanol precipitation of CWE presented remarkable promoting effects on the sporulation of A. cinnamomea, and the effect of LFE was significantly more pronounced than that of YZE, indicating that both LFE and YZE contained compounds that promote the sporulation of A. cinnamomea. Subsequently, liquid chromatography-mass spectrometry (LC-MS) was used to analyze the chemical components of the fractions obtained from CWE, and erythritol was considered as the major component that promotes the sporulation of A. cinnamomea. Finally, the effect of erythritol with a purity of 98% on the fermentation performance of A. cinnamomead was investigated. The result showed that erythritol did significantly promote the sporulation of A. cinnamomea and increased the spore production by 55.17% compared with the control group at the optimal concentration of 1.0 μg/mL. Meanwhile, 1.0 μg/mL erythritol significantly promoted the mycelial growth and synthesis of intracellular polysaccharides (IPS) of A. cinnamomea in submerged fermentation and increased the biomass and the yield of IPS by 18.65% and 260.13%, respectively. However, erythritol had no significant effect on the synthesis of triterpenes in A. cinnamomea.

Smoked pork and chicken sausages were prepared using three different time-temperature combinations; 60 °C for 4 hours, 75 °C for 3 hours, and 90 °C for 2 hours. The amount of polycyclic aromatic hydrocarbons (PAHs) present in these smoked products was estimated using liquid chromatography-mass spectrometry (LC-MS). The PAH compounds were identified using multiple reaction monitoring (MRM) and the retention time of that particular compound. The most potent carcinogenic PAH compound, benzo(a)pyrene (BaP), was not detected in any of the treatments analyzed. The pork and chicken sausages were found to contain the following PAH compounds: Flu, BaA, Chr, BbF, and BkF. Chrysene was found in all samples in the range from 27.38 to 45.08 µg/kg. It was observed that a higher smoking temperature (90 °C) resulted in more PAH compounds in the sausages. Furthermore, the chicken sausages were found to have higher concentrations of PAHs, particularly Chr and BaA, than the pork sausages.

Risperidone is useful for the treatment of schizophrenia symptoms; however, it also has side effects, and an overdose can be harmful. The metabolic effects of risperidone at high therapeutic doses and its metabolites have not been elucidated. Endogenous cellular metabolites may be comprehensively analyzed using untargeted metabolomics-based liquid chromatography-mass spectrometry (LC-MS), which can reveal changes in cell regulation and metabolic pathways. By identifying the metabolites and pathway changes using a nontargeted metabolomics-based LC-MS approach, we aimed to shed light on the potential toxicological effects of high-dose risperidone on brain microvascular endothelial cells (MVECs) associated with the human blood brain barrier. A total of 42 metabolites were selected as significant putative metabolites of the toxicological response of high-dose risperidone in MVECs. Six highly correlated pathways were identified, including those involving diacylglycerol, fatty acid, ceramide, glycerophospholipid, amino acid, and tricarboxylic acid metabolism. We demonstrated that methods focused on metabolomics are useful for identifying metabolites that may be used to clarify the mechanism of drug-induced toxicity.

Biotherapeutic's higher order structure (HOS) is a critical determinant of its functional properties and conformational relevance. Here, we evaluated two covalent labeling methods: diethylpyrocarbonate (DEPC)-labeling and fast photooxidation of proteins (FPOP), in conjunction with mass spectrometry (MS), to investigate structural modifications for the new class of immuno-oncological therapy known as bispecific antigen-binding biotherapeutics (BABB). The evaluated techniques unveiled subtle structural changes occurring at the amino acid residue level within the antigen-binding domain under both native and thermal stress conditions, which cannot be detected by conventional biophysical techniques, e.g., near-ultraviolet circular dichroism (NUV-CD). The determined variations in labeling uptake under native and stress conditions, corroborated by binding assays, shed light on the binding effect, and highlighted the potential of covalent-labeling methods to effectively monitor conformational changes that ultimately influence the product quality. Our study provides a foundation for implementing the developed techniques in elucidating the inherent structural characteristics of novel therapeutics and their conformational stability.

Brain is most cholesterol-rich organ in the body. Since cholesterol does not cross the blood brain barrier, its metabolism is provided in situ by astrocytes and neurons, and it is crucial for maintaining sterol levels and neuronal integrity and function. Recent studies have shown that the levels of cholesterol precursors and metabolites are lower in the brains of animal models of Huntington's disease (HD) while reduced levels of its catabolite are detected in the plasma of patients. In this study, we introduce a novel analytical method designed to fulfill the complex analytical requirements associated with cholesterol metabolites detection in neurodegenerative disorders. The method allows for the simultaneous quantification of a specific set of oxysterols along with cholesterol precursors in biological samples.The proposed method uses an Ultra-High-Performance Liquid Chromatography-Mass Spectrometry (UHPLC-MS) system operating in multiple reaction monitoring (MRM). Since sterols can be found in biological matrices in either free form or esterified to various fatty acids, a three-step extraction procedure was devised, consisting of alkaline hydrolysis, liquid-liquid extraction and final concentration omitting the need for a solid-phase extraction (SPE) step.The validated method achieved a detection limit of 10 ng/mL in plasma and 1 ng/mg in brain tissue, reaching a comparable sensitivity to previously published LC-MS and GC–MS methods. All target analytes were separated on a reverse-phase column employing a segmented gradient and a temperature ramp. This strategy enabled the elution and separation of all selected metabolites within a 30-minutes timeframe. This innovative approach was employed to quantify cholesterol metabolites in both plasma and brain samples from wild-type (WT) and R6/2 mice, a mouse model of HD. The results obtained from the sample analysis highlighted a significant reduction in desmosterol levels in the R6/2 brain at 12 weeks.In conclusion, the proposed method paves the way for further development of high-sensitive and reproducible protocols to comprehensively investigate simultaneous alterations in both cholesterol biosynthesis and catabolism in HD samples.

Background: The fibrous root of ginseng (GFR) is the dried thin branch root or whisker root of Ginseng (Panax ginseng C. A. Mey). It is known for its properties such as tonifying qi, producing body fluid, and quenching thirst. Clinically, it is used to treat conditions such as cough, hemoptysis, thirst, stomach deficiency, and vomiting. While GFR and Ginseng share similar metabolites, they differ in their metabolites ratios and efficacy. Furthermore, the specific role of GFR in protecting the body remains unclear.Methods: We employed ultra-high performance liquid chromatography-triple quadrupole mass spectrometry to examine alterations in brain neurotransmitters and elucidate the impact of GFR on the central nervous system. Additionally, we analyzed the serum and brain metabolic profiles of rats using ultra-high performance liquid chromatography-quadrupole-orbitrap mass spectrometry to discern the effect and underlying mechanism of GFR in delaying aging in naturally aged rats.Results: The findings of the serum biochemical indicators indicate that the intervention of GFR can enhance cardiovascular, oxidative stress, and energy metabolism related indicators in naturally aging rats. Research on brain neurotransmitters suggests that GFR can augment physiological functions such as learning and memory, while also inhibiting central nervous system excitation to a certain degree by maintaining the equilibrium of central neurotransmitters in aged individuals. Twenty-four abnormal metabolites in serum and seventeen abnormal metabolites in brain could be used as potential biomarkers and were involved in multiple metabolic pathways. Among them, in the brain metabolic pathways, alanine, aspartate and glutamate metabolism, arginine and proline metabolism, histidine metabolism, and tyrosine metabolism were closely related to central neurotransmitters. Butanoate metabolism improves energy supply for life activities in the aging body. Cysteine and methionine metabolism contributes to the production of glutathione and taurine and played an antioxidant role. In serum, the regulation of glycerophospholipid metabolism pathway and proline metabolism demonstrated the antioxidant capacity of GFR decoction.Conclution: In summary, GFR plays a role in delaying aging by regulating central neurotransmitters, cardiovascular function, oxidative stress, energy metabolism, and other aspects of the aging body, which lays a foundation for the application of GFR.

In order to investigate the effect of cultivar and processing technique on the chemical composition of tea, this study analyzed green tea, fresh-scent oolong tea, strong-scent oolong tea and black tea made from the fresh leaves of the ‘Tieguanyin’ and ‘Shuixian’ cultivars from the same graphical origin using metabolomics based on ultra-high performance liquid chromatography-time-of-flight mass spectrometry (UPLC-TOF-MS). The results showed that the chemical compositions of tea samples made from different tea cultivars differed considerably. The contents of most lipids, alkaloids and methylated catechins were higher and the contents of theanine, most catechins, flavone (flavonol) glycosides, phenolic acids and N-ethyl-2-pyrrolidinone-substituted flavan-3-ols were lower in ‘Tieguanyin’ tea than in ‘Shuixian’ tea. Processing techniques also significantly impacted the chemical composition of tea. A total of 127 differential compounds were selected, among which the contents of most catechins, dimeric catechins, alkaloids and some amino acid compounds were significantly higher in green tea than in other kinds of tea. The contents of some amino acids and flavone (flavonol) glycosides (quercetin-3-galactoside, and kaempferol-3-glucosylrutanoside) were higher in fresh-scent oolong tea. The contents of N-ethyl-2-pyrrolidinone-substituted flavan-3-ols (EPSF), some flavone (flavonol) glycosides (quercetin-3-glucoside, and kaempferol-3-6”-acetylgalactoside), and lipid compounds in strong-scent oolong tea were higher, while black tea was richer in theaflavins and several flavone (flavonol) glycosides.

BackgroundAtmospheric fine particulate matter (PM2.5) can disrupt the metabolic homeostasis of the liver and accelerate the progression of liver diseases, but there are few studies on the effects of sub-chronic PM2.5 exposure on the liver metabolome. ObjectivesTo investigate the effects of sub-chronic exposure to concentrated PM2.5 on hepatic metabolomics in mice by liquid chromatography-mass spectrometry (LC-MS), and to identify potentially affected metabolites and metabolic pathways. MethodsTwelve male C57BL/6J (6 weeks old) mice were randomly divided into two groups: a concentrated PM2.5 exposure group and a clean air exposure group. The mice were exposed to concentrated PM2.5 using the "Shanghai Meteorological and Environmental Animal Exposure System" at Fudan University. The exposure duration was 8 h per day, 6 d per week, for a total of 8 weeks. The mice's liver tissues were collected 24 h after the completion of exposure. LC-MS was performed to assess changes in the hepatic metabolome. Orthogonal partial least squares discriminant analysis and t-test were employed to identify differentially regulated metabolites between the two groups under the conditions of variable important in projection (VIP)≥1.0 and P

Abstract Background Abiotic stresses in plants include all the environmental conditions that significantly reduce yields, like drought and heat. One of the most significant effects they exert at the cellular level is the accumulation of reactive oxygen species, which cause extensive damage. Plants possess two mechanisms to counter these molecules, i.e. detoxifying enzymes and non-enzymatic antioxidants, which include many classes of specialized metabolites. Sunflower, the fourth global oilseed, is considered moderately drought resistant. Abiotic stress tolerance in this crop has been studied using many approaches, but the control of specialized metabolites in this context remains poorly understood. Here, we performed the first genome-wide association study using abiotic stress-related specialized metabolites as molecular phenotypes in sunflower. After analyzing leaf specialized metabolites of 450 hybrids using liquid chromatography-mass spectrometry, we selected a subset of these compounds based on their association with previously known abiotic stress-related quantitative trait loci. Eventually, we characterized these molecules and their associated genes. Results We putatively annotated 30 compounds which co-localized with abiotic stress-related quantitative trait loci and which were associated to seven most likely candidate genes. A large proportion of these compounds were potential antioxidants, which was in agreement with the role of specialized metabolites in abiotic stresses. The seven associated most likely candidate genes, instead, mainly belonged to cytochromes P450 and glycosyltransferases, two large superfamilies which catalyze greatly diverse reactions and create a wide variety of chemical modifications. This was consistent with the high plasticity of specialized metabolism in plants. Conclusions This is the first characterization of the genetic control of abiotic stress-related specialized metabolites in sunflower. By providing hints concerning the importance of antioxidant molecules in this biological context, and by highlighting some of the potential molecular mechanisms underlying their biosynthesis, it could pave the way for novel applications in breeding. Although further analyses will be required to better understand this topic, studying how antioxidants contribute to the tolerance to abiotic stresses in sunflower appears as a promising area of research.

Abstract Background Clozapine is a highly effective second-generation antipsychotic with few extrapyramidal reactions, making it a preferred choice among clinicians. However, instances of acute clozapine poisoning resulting from suicide attempts and misuse have been reported. Through our review of existing literature, we identified that we believe to be the highest recorded overdose of clozapine in elderly patients, resulting in a nonfatal outcome. Case presentation The case report involves a 71-year-old female with a history of depression who ingested a dose of 10,000 mg of clozapine. Approximately 6 h after the overdose, the clozapine level was 5,200 μg/L, significantly surpassing the recommended therapeutic concentration range of 350–600 μg/L. After gastric lavage and hemoperfusion, the blood level dropped to 1847.11 μg/L. Notably, during therapeutic drugs monitoring (TDM), we found a perplexing spike in the patient’s blood level to 5554.15 μg/L after the second hemoperfusion. Conclusion In this case we mainly focused on the abnormal fluctuations in the concentration of clozapine. We conducted a comprehensive analysis of potential factors contributing to this abnormal phenomenon in terms of the patient’s age, clinical symptoms, various laboratory test indexes, and the pharmacokinetics of clozapine. Our findings underscore the importance of timely TDM and the precision of results in managing elderly patients experiencing high-dose clozapine poisoning.

In this study, the digestion-promoting function of an aqueous extract from Maojian green tea extract (MJ-GTE) was evaluated by small intestinal motility in mice as well as body mass, body mass gain, food intake, food utilization rate, gastric pepsin activity, and gastric pepsin excretion in rats. The chemical composition of MJ-GTE was then systematically analyzed using metabolomics based on ultra-high performance liquid chromatography-quadrupole electrostatic orbitrap mass spectrometry (UPLC-Q-Exactive/MS). The results of animal experiments showed that the intestinal propulsion ratio of ink in the high-dose MJ-GTE group (0.83 g/(kg·d)) was significantly increased compared with the model group (P < 0.05), and gastric pepsin excretion in the medium-dose MJ-GTE group (0.21 g/(kg·d)) was significantly increased compared with the negative control group (deionized water) (P < 0.05), which collectively indicated that MJ-GTE has a digestion-promoting effect. The metabolomics analysis identified 98 compounds, among which, flavones (apigenin and luteolin, 0.14–0.77 mg/g), flavanones (naringenin and eriodictyol, 0.49–1.49 mg/g), flavone-7-O-glycosides (0.57–9.07 mg/g), and flavanone-7-O-glycosides (4.49–38.98 mg/g) were the major components in MJ-GTE. This study will provide a theoretical basis for the promotion and development of Maojian green tea and related products in the future.

Diabetic cardiomyopathy (DCM) is a serious complication of type 2 diabetes mellitus (T2DM) that contributes to cardiovascular morbidity and mortality. However, the metabolic alterations and specific biomarkers associated with DCM in T2DM remain unclear. In this study, we conducted a comprehensive metabolomic analysis using liquid chromatography–mass spectrometry (LC-MS) to investigate the plasma metabolite profiles of T2DM patients with and without DCM. We identified significant differences in metabolite levels between the groups, highlighting the dysregulation of various metabolic pathways, including starch and sucrose metabolism, steroid hormone biosynthesis, tryptophan metabolism, purine metabolism, and pyrimidine metabolism. Although several metabolites showed altered abundance in DCM, they also shared characteristics of DCM and T2DM rather than specific to DCM. Additionally, through biomarker analyses, we identified potential biomarkers for DCM, such as cytidine triphosphate, 11-ketoetiocholanolone, saccharopine, nervonic acid, and erucic acid. These biomarkers demonstrated distinct patterns and associations with metabolic pathways related to DCM. Our findings provide insights into the metabolic changes associated with DCM in T2DM patients and highlight potential biomarkers for further validation and clinical application. Further research is needed to elucidate the underlying mechanisms and validate the diagnostic and prognostic value of these biomarkers in larger cohorts.

Mass spectrometry (MS) has revolutionized clinical chemistry, offering unparalleled capabilities for biomolecule analysis. This review explores the growing significance of mass spectrometry (MS), particularly when coupled with liquid chromatography (LC), in identifying disease biomarkers and quantifying biomolecules for diagnostic and prognostic purposes. The unique advantages of MS in accurately identifying and quantifying diverse molecules have positioned it as a cornerstone in personalized-medicine advancement. MS-based technologies have transformed precision medicine, enabling a comprehensive understanding of disease mechanisms and patient-specific treatment responses. LC-MS has shown exceptional utility in analyzing complex biological matrices, while high-resolution MS has expanded analytical capabilities, allowing the detection of low-abundance molecules and the elucidation of complex biological pathways. The integration of MS with other techniques, such as ion mobility spectrometry, has opened new avenues for biomarker discovery and validation. As we progress toward precision medicine, MS-based technologies will be crucial in addressing the challenges of individualized patient care, driving innovations in disease diagnosis, prognosis, and treatment strategies.

Pesticide application is used in horticulture to reduce plant damage from organisms such as insects and mites. Systemic insecticides are highly efficacious and readily taken up by plant tissues. However, pesticide-treated plants may impose risks to nontarget insects or other organisms within ecosystems. In this study, insecticide residues in nectar, leaves, and flower petals of the horticulturally significant herbaceous annual snapdragon, Antirrhinum majus (Lamiales: Plantaginaceae), were assessed at two locations over several weeks following foliar and drench treatment with five systemic insecticides. Concentrations of the insecticides were determined by liquid chromatography–mass spectrometry. The independent effects Application Method, Application Rate, and Time were statistically significant among all active ingredients in the three matrices in both sites in California (CA) and New Jersey (NJ). The interaction effects were also generally statistically significant in the CA site but less consistently so in the NJ site, dependent on the active ingredient and matrix. Post hoc analyses found the highest residue concentrations in leaves and the lowest in nectar, a trend generally consistent over time regardless of active ingredient for both the CA and NJ sites. The results of this study are discussed in the context of conserving pollinators and other beneficial insects. It is recommended that similar studies should be implemented in different geographical regions and climates, along with multiyear studies for perennial ornamental plants.

Globally, the presence of cyanotoxins in water supplies and food has been widely investigated for over a decade. Cyanotoxins are harmful metabolites produced by toxic cyanobacterial genera. These metabolites belong to diverse chemical classes, with a variety of physicochemical properties, chemical structures, and toxic activities. The present study seeks to investigate the occurrence of cyanotoxins in water supplies destined for food processing and assess the human health risk from exposure to cyanotoxins. To achieve this, a simple, sensitive, and reliable analytical method was developed for the determination of microcystins (MC-RR, MC-LR, MC-YR) in process water, raw maize meal, and cooked maize (porridge) at ppb (parts per billion) levels. These compounds were extracted using Solid Phase Extraction (SPE) with optimized parameters; thereafter, Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS) was used for the rapid determination of the analytes selected for the present study. The method developed was applied to samples collected from the meal grinding station located in Mawoni village in South Africa; and was able to detect and quantify all the target cyanotoxins. MC-LR, MC-YR and MC-RR were detected at concentrations ranging from 10 to 11.2 μg/L, 9.1–9.4 μg/L, and 2.3–3.5 μg/L, in water samples, respectively. However, MC-YR was not detected in ground water sample. Moreover, MC-LR, MC-YR, and MC-RR concentrations in maize and porridge samples ranged between 9.2 and 11.2, 5.5–8.6, and 6.3–9.3 μg/kg dry weight, respectively. The hazard quotient index (HQi) levels found in the present study ranged between 2.2 – 8.4 and 0.11–8.9 for adults and children, respectively, representing potential risks to human health. Findings from LC-MS/MS reveal that cyanotoxins can be transferred from water to food during food processing using cyanotoxins contaminated water. Furthermore, the methods developed can be used by environmental and health agencies to strengthen the monitoring of cyanotoxins in water and food.

IntroductionWhile known animal exposures to human “drugs of abuse” (DA) were previously considered relatively uncommon in veterinary medicine, the trends are changing. Marijuana and amphetamines are among the 20 toxicants most frequently consulted about with the Pet Poison Helpline. When such exposures occur, they are typically considered emergencies.MethodsThis retrospective study describes confirmed cases of DA exposure in pets from the California Animal Health and Food Safety Laboratory System (CAHFS), 2013–2023.ResultsFifty-seven samples tested positive for DA through liquid chromatography with tandem mass spectrometry analysis (qualitative method). In 75% (43/57) of the DA screen tests, the detected drugs included amphetamine-type stimulants and metabolites (methamphetamine, amphetamine, or both). In 47% (27/57) of cases, a combination of more than one drug group was found. Most cases were diagnosed from a urine specimen. In at least 32% (18/57) of cases, the samples were submitted due to suspicions of animal cruelty, and at least 41% (23/57) of the patients were deceased when the samples were submitted.DiscussionMore studies on the prevalence of illicit drugs in small animals, using confirmatory testing, are warranted to fully understand the significance of this emerging toxicological hazard in veterinary medicine.

Amino-containing compounds, including amino acids, aliphatic amines, aromatic amines, small peptides and catecholamines, are involved in various biological processes and play vital roles in multiple metabolic pathways. Previous studies indicated that some amino-containing metabolites are significant diagnostic and prognostic biomarkers of gastric cancer. However, the discovery of precise biomarkers for the preoperative diagnosis of gastric cancer is still in an urgent need. Herein, we established a polarity-regulated derivatization method coupled with liquid chromatography-mass spectrometry (LC-MS) for amino-containing metabolites profiling in the serum samples of patients with gastric cancer and healthy controls, based on our newly designed and synthesized derivatization reagent (S)-3-(1-(diisopropoxyphosphoryl) pyrrolidine-2-carboxamido)-N-hydroxysuccinimidyl ester (3-DP-NHS). Enhanced separation efficiency and detection sensitivity for amino-containing metabolites were achieved after derivatization. This method exhibited good linearity, recovery, intra- and inter-day precision and accuracy. Only 5 μL serum is needed for untargeted analysis, enabling 202 amino-containing metabolites to be detected. Statistical analysis revealed altered amino acid metabolisms in patients with gastric cancer. Furthermore, ultra high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS/MS) analysis quantification revealed increased serum levels of tryptamine and decreased concentrations of arginine and tryptophan in patients with gastric cancer. Receiver operating characteristic (ROC) curves indicated that an increased tryptamine/tryptophan ratio could serve as a potential biomarker for gastric cancer diagnosis. This study demostrated the possibility of using serum amino acid biomarkers for gastric cancer diagnosis, providing new avenues for the treatment of gastric cancer.

Abstract Background The brain-gut axis has gained increasing attention due to its contribution to the etiology of various central nervous system disorders. This study aims to elucidate the hypothesis that schizophrenia is associated with disturbances in intestinal microflora and imbalance in intestinal metabolites. By exploring the intricate relationship between the gut and the brain, with the goal of offering fresh perspectives and valuable insights into the potential contribution of intestinal microbial and metabolites dysbiosis to the etiology of schizophrenia. Methods In this study, we used a 16S ribosomal RNA (16S rRNA) gene sequence–based approach and an untargeted liquid chromatography-mass spectrometry-based metabolic profiling approach to measure the gut microbiome and microbial metabolites from 44 healthy controls, 41 acute patients, and 39 remission patients, to evaluate whether microbial dysbiosis and microbial metabolite biomarkers were linked with the severity of schizophrenic symptoms. Results Here, we identified 20 dominant disturbances in the gut microbial composition of patients compared with healthy controls, with 3 orders, 4 families, 9 genera, and 4 species. Several unique bacterial taxa associated with schizophrenia severity. Compared with healthy controls, 145 unusual microflora metabolites were detected in the acute and remission groups, which were mainly involved in environmental information processing, metabolism, organismal systems, and human diseases in the Kyoto encyclopedia of genes and genomes pathway. The Sankey diagram showed that 4 abnormal intestinal and 4 anomalous intestinal microbial metabolites were associated with psychiatric clinical symptoms. Conclusions These findings suggest a possible interactive influence of the gut microbiota and their metabolites on the pathophysiology of schizophrenia.

Abstract Background Individuals with sepsis exhibited a higher likelihood of benefiting from early initiation of specialized treatment to enhance the prognosis of the condition. The objective of this study is to identify potential biomarkers of sepsis by means of serum metabolomics. Materials and methods The screening of putative biomarkers of sepsis was conducted using serum samples from patients with sepsis and a control group of healthy individuals. The pathogenesis of sepsis was determined through the utilization of liquid chromatography-mass spectrometry-based metabolic profiles and bioinformatic techniques, which in turn provided a foundation for timely diagnosis and intervention. Results Individuals with sepsis had significantly different metabolic characteristics compared to those with normal health. The concentrations of phosphatidylcholines (PCs), phosphatidylserine (PS), lysophosphatidylethanolamine (LysoPEs), and lysophosphatidylcholine (LysoPCs) exhibited a decrease, while the levels of creatinine, C17-Sphinganine, and PS(22:0/22:1(11Z)) demonstrated an increase in the serum of sepsis patients when compared to the control group. Additionally, ROC curves were generated to assess the discriminatory ability of the differentially expressed metabolites. The area under the ROC curve for PS (22:0/22:1(11Z)) and C17-Sphinganine were determined to be 0.976 and 0.913, respectively. These metabolites may potentially serve as diagnostic markers for sepsis. Additionally, the pathogenesis of sepsis is associated with mTOR signaling, NF-κB signaling pathway, calcium signaling, calcium transport, and tRNA charging pathway. Conclusion The identification of differential expression of these metabolites in sepsis serum samples could aid in the timely diagnosis and intervention of sepsis, as well as enhance our understanding of its pathogenesis.

Objective: To screen for tea peptides with inhibitory activity against α-glucosidase. Methods: The response surface method was used to optimize the preparation process of tea peptides. Affinity ultrafiltration was used to isolate tea peptides that bind with α-glucosidase, and liquid chromatography-mass spectrometry was used to determine the sequence of the isolated peptides. Virtual screening was performed using bioinformatics methods. Results: The optimal preparation process of tea leaf enzymatic hydrolysis products was alkaline protease hydrolysis temperature of 50 ℃, enzymatic hydrolysis time of 3 h, and a liquid-to-solid ratio of 10:1 (mL/g). The α-glucosidase inhibitory rate was 57.29%. From this, 624 peptide segments were identified, and LIGF was selected for its α-glucosidase inhibitory activity. At a concentration of 5 mg/mL, LIGF exhibited a maximum inhibition rate of 88.13% against α-glucosidase and an IC50 value of 1.22 mg/mL. Molecular docking showed that LIGF could form 5 hydrogen bonds with α-glucosidase, and the binding energy was −3.51 kJ, indicating a high affinity, stability and ability to bind to α-glucosidase. Conclusion: LIGF had potential value as a therapeutic drug for type II diabetes.

ABSTRACTThe bioactive constituents of Nettle (Urtica dioica L.) plants were extracted by green and environmentally friendly methods using several deep eutectic solvents (DESs) followed by liquid chromatography-mass spectrometry separation and identification. To optimize the extraction process, bio-solvents composed of organic acids such as hydrogen bond donors (HBD) and mannitol/menthol/sugars as HBA were selected and coupled with ultrasound-assisted extraction. The maximum bioactive compound contents were TPC 2423 mg GAE/100 g, TFC 134.71 mg CE/100 g, and DPPH 1071.05 mol TE/mL. The optimum extraction parameters to achieve the highest values of phenolic compounds from Nettle (Urtica dioica L.) via UAE-DES were obtained at an extraction temperature 70°C, extraction time of 30 min, and water addition on DES 20%. The ideal experimental conditions were confirmed using the Central Composite Design (CCD) of the Response Surface Methodology (RSM) model to analyze the significance of extraction parameters demonstrating high prediction levels of the bioactive compounds. Liquid chromatography coupled with diode array detection and electrospray ionization tandem mass spectrometry (LC-DAD-ESI-MS/MS) was used to identify the extracts. The results showed that quercetin-3-O-rutinoside, 5-ocaffeoylquinic acid, and caffeoyl malic acid were the three compounds with the highest abundance. For the first time, the extraction of bioactive compounds from nettle using a food-grade DES that contains GRAS components is reported. According to these findings, the best solvent for extracting bioactive substances for food-grade applications is DESCAMAL (citric acid/maltose).

Hypoxic–ischemic encephalopathy (HIE) is a severe neurological disorder caused by perinatal asphyxia with significant consequences. Early recognition and intervention are crucial, with therapeutic hypothermia (TH) being the primary treatment, but its efficacy depends on early initiation of treatment. Accurately assessing the HIE severity in neonatal care poses challenges, but omics approaches have made significant contribution to understanding its complex pathophysiology. Our study further explores the impact of HIE on the blood metabolome over time and investigated changes associated with hypothermia’s therapeutic effects. Using a rat model of hypoxic–ischemic brain injury, we comprehensively analyzed dried blood spot samples for fat-soluble compounds using HPLC-MS. Our research shows significant changes in the blood metabolome after HIE, with a particularly rapid recovery of lipid metabolism observed. Significant changes in lipid metabolites were observed after 3 h of HIE, including increases in ceramides, carnitines, certain fatty acids, phosphocholines, and phosphoethanolamines, while sphingomyelins and N-acylethanolamines (NAEs) decreased (p < 0.05). Furthermore, NAEs were found to be significant features in the OPLS-DA model for HIE diagnosis, with an area under the curve of 0.812. TH showed a notable association with decreased concentrations of ceramides. Enrichment analysis further corroborated these observations, showing modulation in several key metabolic pathways, including arachidonic acid oxylipin metabolism, eicosanoid metabolism via lipooxygenases, and leukotriene C4 synthesis deficiency. Our study reveals dynamic changes in the blood metabolome after HIE and the therapeutic effects of hypothermia, which improves our understanding of the pathophysiology of HIE and could lead to the development of new rapid diagnostic approaches for neonatal HIE.

The meat of local livestock breeds often has unique qualities and flavors. In this study, three Shanghai native pig breeds (MSZ, SWT, and SHB) exhibited better meat quality traits than globalized commercial pig breeds (DLY). Subsequently, metabolomic and lipidomic differences in the longissimus dorsi (L) and gluteus (T) muscles of the Shanghai native pig breeds and DLY pig breed were compared using liquid chromatography–mass spectrometry (LC–MS). The results demonstrated that the metabolites mainly consisted of (28.16%) lipids and lipid-like molecules, and (25.87%) organic acids and their derivatives were the two most dominant groups. Hundreds of differential expression metabolites were identified in every compared group, respectively. One-way ANOVA was applied to test the significance between multiple groups. Among the 20 most abundant differential metabolites, L-carnitine was significantly different in the muscles of the four pig breeds (p-value = 7.322 × 10−11). It was significantly higher in the L and T muscles of the two indigenous black pig breeds (MSZ and SWT) than in the DLY pigs (p-value < 0.001). Similarly, lipidomic analysis revealed the PA (18:0/18:2) was significantly more abundant in the muscle of these two black breeds than that in the DLY breed (p-value < 0.001). These specific metabolites and lipids might influence the meat quality and taste properties and lead to customer preferences. Therefore, this study provided insights into the characterization of meat metabolites and lipids in Shanghai native pig breeds.

Cold chain disruption during logistics is a critical cause of food quality degradation. This study simulated 3 possible cold chain patterns and combined liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-MS (GC-MS) to explore the quality deterioration and metabolic changes of fresh beef under cold chain disruption. The findings revealed that the cold chain interruption led to an upregulation of metabolites associated with lipid metabolism and protein degradation, which are linked to severe quality deterioration. Specifically, L-valine, L-isoleucine, β-alanine, 12-ketoeicosatetraenoic acid, and 5,6-epoxyeicosatrienoic acid exhibited upregulation. Tricarboxylic acid cycle intermediate metabolites oxoglutaric acid is a pivotal up-regulated metabolite that links protein degradation processes and post-slaughter energy production. Additionally, carbonyl compounds showed significant alterations in the beef under cold chain disruptions. Moreover, the later stage of cold chain disruption triggered a rapid collapse of beef quality. This study combined GC-MS and LC-MS to identify key metabolites associated with quality deterioration caused by cold chain disruption, providing a new perspective on beef quality monitor in cold logistics circulation.

This study investigated the ability of aqueous leaf extracts of Heliotropium bacciferum (HE), Ocimum dhofarense (OE), and Zataria multiflora (ZE) to detoxify aflatoxin B1 (AFB1) under in vitro and in vivo conditions. The results showed that HE, OE, and ZE degraded 95%, 93%, and 92% of AFB1, respectively, after 72 h incubation at 37 °C. The degradation of AFB1 was validated by liquid chromatography–mass spectrometry analysis. A molecular ion peak at m/z 313 specific to AFB1 (C17H12O6) was observed in the mass spectrum of untreated AFB1 (control). However, the level of AFB1 was decreased to untraceable levels in response to treatment with these plant extracts. HE, OE, and ZE effectively detoxified AFB1 in a concentration-dependent manner, resulting in mortality rates of 65, 70, and 75% of brine shrimp, respectively, in contrast to 90% in the untreated AFB1 (control). The hepatoprotective effect of HE, OE, and ZE against AFB1-induced oxidative stress pathogenesis was investigated using mice as an experimental model. Glutathione depletion, impairment of total antioxidant capacity, and increase in DNA oxidative damage were observed in liver tissues of mice treated with AFB1. However, HE, OE, and ZE extract supplementation suppressed the oxidative damage associated with AFB1 treatment. Our findings indicated that HE, OE, and ZE were highly effective in the detoxification of AFB1. In addition, HE, OE, and ZE act as potent antioxidants and combat the AFB1-associated oxidative stress and liver pathogenesis, suggesting that these plants might be valuable for the development of functional foods aimed at minimizing the toxic effects of AFB1.

Factors influencing the sour taste of coffee and the properties of chlorogenic acid are not yet fully understood. This study aimed to evaluate the impact of roasting degree on pH-associated changes in coffee bean extract and the thermal stability of chlorogenic acid. Coffee bean extract pH decreased up to a chromaticity value of 75 but increased with higher chromaticity values. Ultraviolet–visible spectrophotometry and structural analysis attributed this effect to chlorogenic and caffeic acids. Moreover, liquid chromatography-mass spectrometry analysis identified four chlorogenic acid types in green coffee bean extract. Chlorogenic acid isomers were eluted broadly on HPLC, and a chlorogenic acid fraction graph with two peaks, fractions 5 and 9, was obtained. Among the various fractions, the isomer in fraction 5 had significantly lower thermal stability, indicating that thermal stability differs between chlorogenic acid isomers.

Ulcerative colitis (UC) is an inflammatory bowel disease and associated with metabolic imbalance. Luteolin (LUT) reportedly exhibits anti-inflammatory activity. However, its regulatory effects on metabolites remain indistinct. Here, the effects of LUT on immune response and oxidative stress in UC were determined. Serum metabolomics profiles of UC rats treated with LUT were obtained utilizing liquid chromatography-mass spectrometry. The results revealed that LUT treatment alleviated colon tissue injury, colon shortening, weight loss, and inflammatory response in UC rats. Additionally, the levels of superoxide dismutase and total antioxidant capacity were elevated, but malondialdehyde content was reduced in serum of UC rats, while these changes were abrogated by LUT. Metabolomics analysis unveiled that l-malic acid, creatinine, l-glutamine, and l-lactic acid levels were remarkably decreased, while dimethyl sulfone, 5-methylcytosine, cysteine-S-sulfate, and jasmonic acid levels were notably increased after LUT treatment. Furthermore, differential metabolites primarily participated in d-glutamine and d-glutamate metabolism, glutathione metabolism, and citrate cycle pathways. In summary, these results demonstrated that LUT improved immune response, alleviated oxidative stress, and altered metabolites in UC rats. This study lays the root for further exploring the mechanism of LUT in the treatment of UC.

Abstract Degeneration of the intervertebral disc is an age‐related condition. It also accompanies the disappearance of the notochordal cells, which are remnants of the developmental stages of the nucleus pulposus (NP). Molecular changes such as extracellular matrix catabolism, cellular phenotype, and glycosaminoglycan loss in the NP have been extensively studied. However, as one of the most significant co‐ and posttranslational modifications, glycosylation has been overlooked in cells in degeneration. Here, we aim to characterize the N‐glycome of young and mature NP and identify patterns related to aging. Accordingly, we isolated N‐glycans from notochordal cell‐rich NP from porcine discs, characterized them using a combined approach of exoglycosidase digestions and analysis with hydrophilic interaction ultra‐performance liquid chromatography and mass spectrometry. We have assigned over 300 individual N‐glycans for each age group. Moreover, we observed a notable abundance of antennary structures, galactosylation, fucosylation, and sialylation in both age groups. In addition, as indicated from our results, increasing outer arm fucosylation and decreasing α(2,3)‐linked sialylation with aging suggest that these traits are age‐dependent. Lastly, we have focused on an extensive characterization of the N‐glycome of the notochordal cell‐rich NP in aging without inferred degeneration, describing glycosylation changes specific for aging only. Our findings in combination with those of other studies, suggest that the degeneration of the NP does not involve identical processes as aging.

Abstract Background Xuelian granule (XL), a traditional Chinese medicine (TCM) formula, has been used for the treatment of diabetic nephropathy for a long time as a hospital preparation. Because the active ingredients in the XL that can help to treat diabetic nephropathy are still unclear, which limits the interpretation for its pharmacological mechanism, further development and subsequent study on the material basis of its efficacy. Methods In this study, a screening method based on inhibition activity against aldose reductase (AR) was employed for activity-directed chemical analysis of XL using ultra-high performance liquid chromatography combined with quadrupole-orbitrap high resolution mass spectrometry (UHPLC-Q-orbitrap-HRMS) technique. Results A total of 178 compounds, including 46 terpenes, 47 organic acids, 25 flavonoids, 29 phenylethanoid glycosides, and 31 other types, were tentatively identified from XL which might responsible for its AR inhibition activity. Conclusion This is the first study for a systematic, rapid, and accurate qualitative analysis of XL. This research provides a scientific and experimental basis for further researches on pharmacodynamics material basis and quality control of XL.

Abstract Tomato (Solanum lycopersicum L.) is one of the most consumed vegetables worldwide. The ripening of tomato is performed for its freshness and represented by color and gene expression. In our previous study, we performed molecular analyses on tomato ripening with and without sound-wave treatment. In the present study, we performed metabolomics analysis of ripening tomatoes with and without sound-wave treatment to expand our knowledge of tomato ripening. To achieve this goal, tomatoes at 7, 10, and 14 d of ripening were selected and analyzed via liquid chromatography–mass spectrometry (LC–MS) and gas chromatography–MS (GC–MS). A total of 33 major metabolites, including 14 LC–MS- and 19 GC–MS-derived metabolites, were assigned based on variable importance projection and p values and subjected to statistical analysis. Apparent morphology and partial least squares–discriminant analysis were consistent with the general ripening process based on color. Moreover, metabolomics analysis showed similar experimental results to those of previous studies. The quantification of metabolites with LC–MS showed decreasing levels of adenosine, tryptophan, and phytosphingosine upon sound-wave treatment. In GC–MS analysis, 4-Aminobutanoic acid and aspartic acid were decreased upon sound-wave treatment. On the other way, the quantity of malic acid, citric acid, and sucrose was increased with the treatment. The findings of this study can assist in the application of sound-wave treatment for delaying ripening in tomatoes and improving their market value.

This study was conducted in order to explore the effect of lead (Pb) stress on the nutritional value and flavor of barley seedling powder. Fourier transform infrared (FTIR) spectroscopy and targeted metabolomics based on liquid chromatography-mass spectrometry (LC-MS) were used for quantitative and qualitative analysis of protein secondary structure and amino acid contents in barley seedling powders with and without Pb stress during growth. The results showed that Pb stress led to the transformation of protein secondary structure in barley seedlings from disordered (β-turn and random coil) to ordered (α-helix) structure. Through high-throughput targeted metabolomic analysis, a total of 25 amino acids were identified and quantified in barley seedling powder. Pb stress decreased the contents of Arg, Pro and His, but increased the content of Val, which proves that Pb could reduce the nutritional value and sweetness, and increase the bitterness of barley seedling powder. These results provide some basic data for the development of barley-based health foods.

Qingwei Geng,1,&ast; Guoqi Wei,2,&ast; Yebei Hu,1 Jinhui Xu,1 Xiuzu Song1 1Department of Dermatology, Hangzhou Third People’s Hospital, Hangzhou, Zhejiang, 310009, People’s Republic of China; 2Hangzhou Fuyang Hospital of Traditional Chinese Medicine, Hangzhou, Zhejiang, 322499, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xiuzu Song, Department of Dermatology, Hangzhou Third People’s Hospital, Affiliated Hangzhou Dermatology Hospital, Zhejiang University School of Medicine, West Lake Road 38, Hangzhou, 310009, People’s Republic of China, Tel +86-0571-87823102, Email songxiuzu@sina.comBackground: The skin barrier is the first line of defense of the body, while skin lipids play an important role in the skin permeability barrier. Lamellar bodies are also involved in maintaining the stability of the skin permeability barrier. However, the exact origin of lamellar bodies remains unclear. Recent studies have suggested that autophagy may participate in the formation of lamellar bodies.Aim: This study aimed to investigate the role of autophagy in the formation of lamellar bodies in keratinocytes and the regulation of keratinocyte lipids.Methods: Keratinocytes were incubated with autophagy inducer Rapamycin and autophagy inhibitor Bafilomycin A1. The changes in autophagy flux were detected by Western blot, and the formation of lamellar bodies was observed by transmission electron microscopy. Furthermore, the changes in keratinocytes lipidomics were detected by liquid chromatography-mass spectrometry.Results: Our research showed that the autophagy inducer promoted autophagy activation and formation of lamellar bodies in keratinocytes, while the inhibitor inhibited autophagy signals and the formation of lamellar bodies in keratinocytes. In addition, the lipidomics results revealed a significant change in glycerophospholipids after autophagy induction and autophagy inhibition.Conclusion: These results demonstrate that autophagy may play an essential role in skin lipids via glycerophospholipids pathway.Keywords: skin barrier, lipid, liquid chromatography-mass spectrometry, rapamycin, Bafilomycin A1, keratinocytes

Toxicology testing is performed in clinical settings, forensic settings, and for controlling doping. Drug screening is a toxicology test to determine if drugs are present in biological samples. The most common specimen type for drug testing is urine, as drugs and/or their metabolites are often more concentrated in the urine, extending the detection window of drugs. The dilute-and-shoot method is a simple procedure used in toxicology testing, where a sample is diluted before being directly injected into the liquid chromatography-mass spectrometry (LC-MS) system. This method is easy, quick, and cost-saving, and can be used for protein-poor liquid specimens such as urine. Thus, it is reasonable and attractive for busy toxicology laboratories to combine the dilute-and-shoot method with high-resolution hyphenated-MS for urine drug screening. This method has several disadvantages, including a suboptimal detection capability for certain analytes, as well as interference from co-eluting matrix components called matrix effects, in which co-eluting matrix molecules alter the ionization efficiency of the analyte molecules at the ionization source in LC-MS, altering (mostly reducing) the analyte detection capability. The matrix effect testing is essential for the validation of LC-MS-based assays. A reasonable approach to addressing these undesirable effects would be to minimize these components. The most straightforward approach is to reduce the amounts of matrix components by using a higher dilution of the specimen and a lower volume for specimen injection. Optimization of the chromatographic separation is another reasonable approach for reducing co-eluting matrix components with the analyte.

Egg production systems use antibiotics to treat and prevent the spread of diseases. Antibiotics remain in the animal's body in their original or metabolized form and they can remain in different organs and eggs. The constant consumption of trace doses of antibiotics through food is a serious public health concern, as it can potentiate antimicrobial resistance (AMR). To protect public health, the European Commission defined recommendations to reduce and control the use of those substances in farm animals. Detection and quantification methods using liquid chromatography-mass spectrometry (LC-MS) allow checking whether the antibiotic content meets the maximum residue limit (MRL) or if prohibited antibiotics are present. The egg is a complex matrix and the antibiotic extraction from this matrix is a challenger step present in the detection and quantification process of these molecules. Therefore, this review presents an overview of methods for extracting antibiotics from eggs, and serves as a baseline for new methods able to extract as many antibiotics as possible, coupled with an accurate detection strategy. According to the information collected, the most common extraction solutions were formic acid with acetonitrile, water, and solutions containing ethylenediaminetetraacetic acid (EDTA). Purification via solid-phase extraction (SPE) is also common. Among the detection methods, chromatography coupled to quadrupole mass detectors with positive electrospray ionization using octadecylsilane (C18) as the stationary phase is the most common approach. The developments achieved to optimized analytical strategies are in line with the needed tools to guarantee food safety monitoring and the fulfillment of regulations in place.

Scientific and technological advancement of the mid- and late twentieth century led to the creation of a large number of new psychoactive substances (NPS), diverse in composition and spectrum of action. The primary stages of fight the spread of NPS are awareness of their characteristics, as well as the possibility of detecting both the substances themselves and biomarkers of their use (metabolites) in human biological objects. The purpose of the work is to review psychoactive compounds and methods of their detection performed for diagnostic purposes. The paper presents brief characteristics of the most common NPS, as well as features of their metabolism in the human. It is shown that the analysis of biological specimens collected from susceptive drug users is difficult due to the low content of analytes, the matrix influences, metabolism and the formation of various artifacts. The general scheme of screening of biological objects assumes the presence of two stages – a preliminary express analysis performed using immunochemical test systems, and a confirmatory analysis performed by gas and liquid chromatography-mass spectrometry. The article discusses the possibilities and limitations of targeted and non-targeted screening. The subject of special consideration was the problems of using chromatography-mass spectrometry methods of NPS analysis in Russian – issues of accessibility of standard substances, search libraries, etc. The article also presents brief characteristics of individual families of NPS, such as stimulants, synthetic cannabinoids, synthetic opioids and hallucinogens. In addition, the article shows that in a number of foreign armies, in particular, in the Armed forces of Ukraine (AFU), drugs and psychostimulants are used to create «fearless soldiers». Thus, methadone, amphetamine and other psychoactive substances, as well as a psychotomimetic – a structural and pharmacological similarity of the prohibited BZ – were found in objects delivered from the positions of the AFU for analysis.

为考察不同产地文冠果种仁黄酮类成分的差异，以山东潍坊的60份和辽宁大连的58份文冠果种仁为原料，采用LC-MS/MS法检测其黄酮类成分及含量，并对两地文冠果种仁黄酮类成分含量的差异进行了对比分析。结果表明：118份文冠果种仁中均检测出5种黄酮类成分，其中芦丁含量为1.422~40.704 μg/g，表没食子儿茶素含量为2.513~6.406 μg/g，没食子儿茶素没食子酸酯含量为1.994~4.574 μg/g，二氢槲皮素含量为1.102~4.452 μg/g，没食子儿茶素含量为0.806~4.057 μg/g；两地文冠果种仁黄酮类成分含量大小依次为芦丁>表没食子儿茶素>没食子儿茶素没食子酸酯>二氢槲皮素>没食子儿茶素；山东潍坊不同样树种仁中黄酮类成分总含量介于10518~25.445 μg/g之间，样树WF927的黄酮类成分总含量最高；辽宁大连不同样树种仁中黄酮类成分的总含量介于8.347~52.087 μg/g之间，样树DL392的黄酮类成分总含量最高；两地文冠果种仁黄酮类成分含量间差异显著，其中辽宁大连文冠果种仁中黄酮类成分总含量平均值（19491 μg/g）极显著高于山东潍坊的（16.320 μg/g）;两地118份文冠果种仁中，黄酮类成分总含量最高的为DL392（52.087 μg/g）、DL395（43.711 μg/g）和DL347（40.481 μg/g）。 In order to investigate the changes of flavonoids in Xanthoceras sorbifolium Bunge kernels from different origins, 60 samples from Weifang, Shandong province and 58 samples from Dalian, Liaoning province were used as raw materials, and the flavonoids composition and contents were detected by LC-MS/MS method, and the differences of flavonoids content from two origins were compared and analyzed. The results showed that 5 kinds of flavonoids were detected in 118 Xanthoceras sorbifolium Bunge kernels, among which the content of rutin was 1.422-40.704 μg/g, the content of epigallocatechin was 2.513-6.406 μg/g, the content of gallocatechingallate was 1.994-4.574 μg/g, the content of dihydroquercetin was 1.102-4.452 μg/g, and the content of gallocatechin was 0.806-4.057 μg/g. Rutin content was the highest, followed by epigallocatechin, gallocatechingallate, dihydroquercetin and gallocatechin. The total flavonoids content of kernels from Weifang, Shandong province was between 10518 μg/g and 25.445 μg/g, and the total flavonoids content of WF927 was the highest, the total flavonoids content of kernels from Dalian, Liaoning province was between 8.347 μg/g and 52.087 μg/g, and the total flavonoids content of DL392 was the highest. The content of flavonoids was significantly different between two origins. The average total flavonoids content in Xanthoceras sorbifolum Bunge kernel from Dalian, Liaoning province (19.491 μg/g) was significantly higher than that from Weifang, Shandong province(16.320 μg/g). 118 Xanthoceras sorbifolium Bunge kernel from two origins had the highest total content of flavonoids in DL392 (52.087 μg/g), DL395 (43.711 μg/g) and DL347 (40481 μg/g).

Aim: In this work, the development for the first time of a green and efficient method to obtain bioactive extracts from Mentha x rotundifolia leaves has been investigated. Methods: The efficiency of three techniques [microwave-assisted extraction (MAE), subcritical water extraction (SWE), and ultrasound-assisted extraction (UAE)] was compared in terms of total phenolic content (TPC) and antioxidant activity [1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2’-azino-bis (3-ethylbenzthioazoline-6-sulfonic acid) (ABTS) assays]. Results: Under similar operating conditions, SWE outperformed MAE and UAE for providing M. rotundifolia extracts with improved antioxidant activity. Further in-depth optimization of the SWE method by means of a Box-Behnken experimental design showed 120°C, 5 min, 0.08 g dry sample: 1 mL water and 2 extraction cycles as optimal experimental parameters to provide the maximum yield of phenolics and the highest bioactivity. The application of the developed SWE method to M. rotundifolia leaves collected in different annuities (2014–2017) showed, in general, no significant differences regarding both composition and antioxidant capacity, as expected from plant samples grown in field under drip irrigation conditions. Conclusions: The SWE method here optimized is shown as a sustainable and efficient alternative for providing bioactive M. rotundifolia extracts with application as functional ingredients, natural preservatives, etc. in the food industry, among others.