From morphology to metabolites: Integrated analysis of chemical and functional diversity in six shape-distinct premium green teas.

Chromatographic determination of antidepressants in plasma and saliva: Towards non-invasive therapeutic monitoring.

Antibody-free quantification of serum infliximab using LC-MS/MS.

Chromatographic separation of dihydroxylated vitamin D3 and accurate quantification of 1α,25(OH)2D3 in human serum.

Efficient host cell protein clearance: A study of membrane adsorbers and resins in biopharmaceutical processes.

Advances and challenges in sample preparation and analytical methods for water-soluble vitamins: Application in food, clinical, pharmaceutical samples.

Pesticide residues in bedroom dust: occurrence, determinants, and health risk assessment.

Pharmacokinetic profile and bioavailability assessment of rubusoside in mouse plasma utilizing UPLC-MS/MS analysis.

Development of a validated and sensitive ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method to simultaneous determination of crizotinib, alectinib and lorlatinib in human plasma.

The genus Lysimachia is of great interest to the scientific community, especially in terms of its potential anticancer effects. In this study, the aerial parts and roots of Lysimachia atropurpurea L. were collected and extracted by maceration using solvents of ethyl acetate (EA), ethanol (EtOH), ethanol/water, and water. The biological activities of the extracts, including antioxidant, enzyme inhibition, and anticancer effects, were evaluated using various assays. High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis revealed a total of 32 compounds in the extracts of L. atropurpurea. The roots showed significantly the highest antioxidant activity compared to the aerial part. In case ofcholinesterase inhibition, the aerial parts of the EtOH extract showed the highest acetylcholinesterase (AChE) inhibition activity, measuring 3.05 mg galatamine equivalent(GALAE)/g. The EtOH and EtOH/water extracts exhibited the strongest cytotoxicity, reducing the viability of human neuroblastoma (SH-SY5Y) and human hepatocarcinoma (HepG2) cancer cells to as low as 4.86-6.33 %. The results of network pharmacology and molecular docking suggest that the extract of L. atropurpurea exertsinhibitory effects on hepatocellular carcinoma through themodulation of SRC, PI3K, and HSP90, while it demonstrates potential inhibitory activity against neuroblastoma by targeting SRC, PI3K, HSP90, ESR1, AKT, and other related targets. In conclusion, the L. atropurpurea extracts showed potential antioxidant, enzyme inhibition, and selective anticancer effects, which support their potential forfurther research as therapeutic agents in drug development.

Human milk exhibits dynamic diurnal variations in bioactive components that are conducive to the consolidation of the biological clock in early life, particularly in the establishment of the sleep-wake cycle in infants. The objectives of the present study are to evaluate the circadian rhythm of amino acids and melatonin in human milk and elucidate their potential sleep-wake regulatory mechanism. Amino acids and melatonin were analyzed in 80 human milk samples collected every 6 hours over a 24-hour period from 20 healthy nursing mothers around 30 days postpartum. Different doses of rhythmic components in human milk were administered to normal mice via oral gavage for 7 days. The comprehensive lab monitoring system (CLAMS) and pentobarbital sodium-induced sleep test (PST) were used to evaluate the sleep-inducing and wake-promoting effects. Liquid chromatography-tandem mass spectrometry and enzyme-linked immunosorbent assay were used to detect the levels of neurotransmitters and hormones to identify the underlying mechanisms. Histidine, phenylalanine, tyrosine, tryptophan and melatonin in human milk exhibit circadian variation with higher levels of histidine, phenylalanine and tyrosine during the daytime and higher contents of tryptophan and melatonin at night. High-dose histidine increased total activity levels in the x-direction and sleep latency and decreased sleep duration through the increased level of histamine and decreased level of gamma-aminobutyric acid (GABA) in the hippocampus and hypothalamus of normal mice. In contrast, high doses of tryptophan and melatonin decreased oxygen consumption rate, x-direction total activity levels and sleep latency and increased sleep duration through different neurotransmitter pathways where high-dose tryptophan increased the 5-hydroxytryptamine level while high doses of melatonin increased melatonin and GABA in the hippocampus and hypothalamus of normal mice. In conclusion, the circadian variation of specific amino acids and melatonin in human milk might contribute to the establishment of the sleep-wake cycle in infants.

Determining the bodily origin of biological traces is a valuable tool in forensic investigations as it helps corroborate testimonies, reconstruct crime-related activities, and select relevant samples for further analysis. Current body fluid identification (BFI) methods rely on enzymatic, spectroscopic, and chemical tests, which are often limited in sensitivity and specificity. Recent research has explored novel markers for BFI, for instance metabolites, based on their potential body fluid/tissue specificity. Metabolites are small molecules produced by human and microbial cellular processes that can be measured using advanced techniques like gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS). These methodologies remain underexplored for the identification of forensically relevant body fluids/tissues. In this pilot study, we employed a high-resolution, untargeted LC-quadrupole time-of-flight (QTOF)-MS approach to investigate body fluid/tissue specific markers from nine biological fluids/tissues, including feces, fingerprick blood, menstrual blood, saliva, semen, skin from palms, urine, vaginal fluid and venous blood. We used sparse partial least-squares discriminant analysis (sPLS-DA) to identify key features responsible for body fluid/tissue-specific clustering and generalized local learning (GLL) to select features directly associated with specific body fluids/tissues. Lastly, we present nine predictive features, one for each fluid/tissue, demonstrating that our approach has the potential to be used in forensic casework.

Balkan endemic nephropathy (BEN) is a chronic kidney disease associated with the consumption of aristolochic acids (AAs) through contaminated food sources. AAs are known to form DNA adducts that are implicated in tumorigenesis and kidney fibrosis. Given the sensitivity of DNA adduct formation to dietary factors, this study aimed to investigate the impact of various dietary practices on AA-DNA adduct formation, thereby assessing the risk of developing BEN. We quantified AA-DNA adducts in DNA extracted from the kidneys and livers of mice subjected to high-fat, high-protein, high-sucrose, and high-salt diets, utilizing a highly sensitive liquid chromatography-tandem mass spectrometry method combined with stable isotope dilution. Our results demonstrated that unbalanced diets significantly elevated the formation of DNA adducts from AAs. Notably, mice fed high-fat diets exhibited increases in adduct levels of 71 and 114% for diets containing 17 and 25% fat, respectively. Mice on a 20% sucrose diet showed an 80% increase in adduct levels compared to those on a standard diet. Further investigations using gut sacs from the small intestines of these mice revealed that the increased level of DNA adduct formation was primarily attributed to enhanced intestinal absorption. Additionally, we observed that drinking alkaline water reduced adduct levels by 30% compared to tap water, likely by decreasing AA absorption. In contrast, commonly used dietary supplements, such as vitamin C and cysteine, significantly increased AA-DNA adduct levels by enhancing the activity of enzymes involved in the metabolic activation of AAs. These findings highlight the critical role of a balanced diet in mitigating the risk of BEN and suggest that alkaline water consumption may serve as a protective strategy for individuals living in AA-contaminated regions.

Periodontitis is the primary cause of tooth loss worldwide. This study aimed to identify herpes simplex virus (HSV) related biomarkers in periodontitis patients. The present study used DIA-based liquid chromatography-tandem mass spectrometry to analyze the proteome of human gingival fibroblasts (HGFs) from one healthy donor across early and late stages (12-72 h) of HSV-1 infection. The study identified 890 differentially expressed proteins. At the early infection stage, there was a notable upregulation of proteins including interferon (IFN) regulatory factor 7, IFN-stimulated genes 15, interleukin 6 (IL6), toll-like receptor 2 (TLR2), and IFN-induced protein (IFI), alongside a downregulation of matrix metalloproteinase 2 (MMP2). We observed the activation of pathways, including the complement and coagulation cascades, lysosome, nucleotide-binding oligomerization domain-like receptors, retinoic acid-inducible gene I-like receptors, and TLR signaling pathways. Conversely, at the late stage, IFIs, IL1, and MMP3 were significantly upregulated, while complement proteins were downregulated. Biomarkers such as TLR2 may underscore the host's antiviral defense response to HSV-1 in the periodontal environment. The present study identified several HGF proteins associated with periodontitis following HSV-1 infection, providing an analytical framework for determining the host's anti-viral defense response to antagonize HSV-1 infection in periodontal tissues.

Adipose tissue has emerged as a prognostic factor in rectal cancer (RC), yet its metabolic role in tumour progression remains poorly understood. This study aims to characterise the metabolic profiles of subcutaneous (SAT) and visceral adipose tissue (VAT) in early and advanced tumours to investigate metabolic alterations that may influence tumour progression. Understanding these alterations could provide insights into mechanisms of rectal cancer and identify therapeutic targets. Global metabolic profiles of 68 SAT and VAT samples from patients with RC were analyzed using high-performance chemical isotope labeling liquid chromatography mass spectrometry. Statistical analyses were stratified by clinical tumour invasion and disease stage. VAT exhibited distinct metabolic signatures between early and advanced tumour invasion. Advanced tumours (cT3-4) had a higher concentration of N-glycolylneuraminic acid and phenylacetylglutamine, whereas early invasive tumours (cT2) exhibited higher levels of dipeptides. Pathway enrichment analyses revealed dysregulations in taurine and hypotaurine metabolism, alanine, aspartate and glutamate metabolism, propanoate metabolism and cellular signalling pathways in advanced invasion. Propanoate metabolism emerged as a key pathway distinguishing early and advanced invasion stages. In SAT, metabolic changes aligned with overall disease stage. Early-stage disease was associated with a higher concentration of dipeptides, whereas advanced stages showed increased N-acetyl-agmatine and N-acetyl-L-noradrenaline. Our study highlights significant metabolic alterations in adipose tissue associated with rectal cancer progression. VAT metabolism reflects tumour invasiveness, while SAT is more indicative of disease stage. Propanoate metabolism may serve as a biomarker for advanced invasion, offering potential for improved staging, reclassification and personalised therapeutic strategies.

For more than 30 years, Qian Ji Sheng Xue Pian (QJSXP) has been used clinically to treat primary immune thrombocytopenia (ITP) with good documented efficacy. However, nothing is known about its underlying mechanisms, effective components, and possible targets. To employ several methodologies to initially investigate the possible targets and therapeutic mechanisms of QJSXP in the treatment of ITP. Liquid chromatography-mass spectrometry (LC-MS) identified the principal chemical elements of QJSXP and assessed its probable active components based on ADME characteristics. The research incorporated multidimensional databases to pinpoint probable targets for the active components. Key pathogenic targets linked to ITP were aggregated from several illness databases, and the STRING and Metascape tools were utilized to examine protein interaction activities and related biological processes. Mendelian randomization (MR) was then utilized to determine beneficial targets for the therapy of ITP. The potential targets, including disease targets and MR-positive targets, were found at the intersection, while risk genes were excluded by heterogeneity, pleiotropy, and Steiger analysis to ascertain the core targets. Molecular docking and molecular dynamics simulations were conducted utilizing Schrodinger and Gromacs software to assess the binding affinity of compound-core targets. The toxicological effects of active molecules targeting critical sites were concurrently anticipated using several toxicity databases. A total of 67 active components and 352 potential targets were discovered in QJSXP, of which 77 were associated with ITP disease targets. Through MR analysis, a total of 12 core genes were identified. Binding scores below - 4.25kcal/mol constituted 82.0%; docking scores below - 5kcal/mol represented 60.1%, with an average binding energy of -5.44kcal/mol. The majority of targets demonstrated strong binding affinity with the components. Toxicity prediction initially highlighted potential hazards, including hepatotoxicity and nephrotoxicity, establishing a foundation for future clinical surveillance. This study has preliminarily identified the active constituents, associated pathways, and possible targets of QJSXP in the treatment of ITP, offering insights for additional experimental validation of QJSXP's mechanism of action in ITP.

Chronic kidney disease (CKD) is a significant global health challenge with a multifaceted etiology. Risk stratification and timely intervention are crucial for mitigating CKD progression. Trimethylamine N-oxide (TMAO), a gut microbiota metabolite, has been implicated in CKD pathogenesis. However, the lack of standardized diagnostic assays for TMAO has limited its clinical applicability. We conducted a multicenter observational study in Southern China. Plasma TMAO levels were quantified using a newly developed liquid chromatography-mass spectrometry (LC-MS)-based diagnostic kit in a cohort of 272 participants, including healthy controls and CKD patients. We analyzed the correlation between TMAO levels and renal function indicators, evaluated the utility of TMAO for risk stratification in CKD, and assessed its independent association with reduced eGFR using multivariable logistic regression. Plasma TMAO levels were significantly elevated in CKD patients compared to healthy controls. TMAO levels increased with CKD severity and exhibited a strong positive correlation with serum creatinine (Scr) and a negative correlation with estimated glomerular filtration rate (eGFR). TMAO levels demonstrated excellent consistency with eGFR in both the full analysis set (FAS) and per protocol set (PPS) populations. Receiver operating characteristic (ROC) curve analysis indicated that TMAO could identify reduced eGFR with high accuracy (AUC: 0.916), which was confirmed by bootstrap validation. After adjusting for age, sex, hypertension, and diabetes, TMAO remained independently associated with reduced eGFR. Our findings suggest that TMAO is a promising and independent complementary biomarker for the risk stratification of CKD, though its prognostic value requires validation in longitudinal studies. The developed LC-MS kit enables robust clinical application. Further research is warranted to validate these findings in larger cohorts and explore TMAO's potential as a therapeutic target for CKD management. Clinical Trails.gov identifier, ChiCTR2400089774 (retrospectively registered in 14/09/2024).

Absolute quantification of polar metabolites using matrix-free calibration curves and post-column infusion of standards in HILIC-MS based metabolomics.

Dysregulated metabolism of ceramides and glycosphingolipids in Parkinson's Disease.

Biological exposure monitoring is particularly useful for understanding skin absorption of hazardous substances; however, existing measurement methods for aromatic amines show room for improvement, as they focus on only unchanged compounds. This study aimed to determine urinary concentrations of the unchanged compounds and metabolites of three aromatic amines (aniline, 2,4-dimethylaniline [m-xylidine], and 2-methylaniline [o-toluidine]) following enzymatic hydrolysis pretreatment, calculate their proportions, and assess markers for occupational biological exposure monitoring. Urine samples were collected at the end of work shifts on consecutive days from 11 workers at an aromatic amine-handling plant. Samples were enzymatically hydrolyzed using glucuronidase and sulfatase. The urinary concentrations of 25 substances were determined using liquid chromatography-tandem mass spectrometry. After exposure to the three aromatic amines, the major urinary excretion types were benzene ring-hydroxylated compounds, ring-hydroxylated and N-acetylated compounds, and side-chain methyl group oxides. The presence of unchanged and N-acetylated compounds was minor. Metabolism and urinary excretion were relatively different between the workers and reported values from animal studies. Excluding metabolites with low specificity, the sum of unchanged compounds and primary metabolites in urine could be useful markers for biological exposure monitoring. Regarding aniline exposure, the relevant markers are the sum of aniline, N-acetyl-4-hydroxyaniline, and 2-hydroxyaniline. For 2,4-dimethylaniline exposure, the markers are the sum of 2,4-dimethylaniline, N-acetyl-4-carboxy-2-methylaniline, and 6-hydroxy-2,4-dimethylaniline. For 2-methylaniline exposure, the markers are the sum of 2-methylaniline, N-acetyl-4-hydroxy-2-methylaniline, 4-hydroxy-2-methylaniline, and 6-hydroxy-2-methylaniline. Urine sampling is recommended at the end of work shifts on consecutive working days.