Search Results (2,558)

Hyperuricemia, characterized by elevated serum uric acid levels, is a major risk factor for gout and kidney disease. This study evaluated the antihyperuricemic effects of Cornus officinalis extract (COE) using urate transporter 1 (URAT1)-expressing oocytes and a hyperuricemia rat model. COE effectively inhibited uric acid absorption by modulating URAT1, with an IC₅₀ value of 3.24 µg/mL. In the hyperuricemia model, COE administration (100 and 200 mg/kg) significantly reduced serum uric acid levels and increased urinary uric acid excretion. The primary constituents of COE, morroniside (MO) and loganin (LO) exerted similar effects, with MO exhibiting potent inhibition of uric acid absorption even at low concentrations. Kidney tissue analysis revealed a reduction in blood urea nitrogen (BUN) levels, indicating improved renal function. Liver function parameters (ALT, AST, and LDH) remained unchanged, suggesting an absence of hepatotoxicity. Ultra-high-performance liquid chromatography with charged aerosol detection (UHPLC-CAD) analysis identified MO (17.8 mg/g), LO (9.8 mg/g), and cornin (1.4 mg/g) as the principal components of COE. These findings suggest that COE enhances uric acid excretion via URAT1 regulation and exerts renoprotective effects, highlighting its potential as an antihyperuricemic agent. Furthermore, MO and LO were identified as the primary active constituents, and COE appears to be a promising therapeutic candidate with a favorable safety profile. Full article

Diamorphine (DIM, heroin) is a semi-synthetic opioid that undergoes rapid conversion to 6-monoacetylmorphine and morphine, producing short-lived biomarkers that are difficult to capture during the process. This review critically explores the evolution of analytical techniques for quantitative DIM analysis in biological matrices from 1980 to 2025. It synthesizes findings across blood, plasma, urine, hair, sweat, and postmortem samples, emphasizing matrix-specific challenges and forensic applicability. Unlike previous opioid reviews that primarily focused on metabolites, this work highlights analytical methods capable of successfully detecting diamorphine itself alongside its key metabolites. This review examines 32 studies spanning three decades and compares three core analytical methods: gas chromatography–mass spectrometry (GC–MS), high-performance liquid chromatography (HPLC) with optical detection and liquid chromatography–mass spectrometry (LC–MS). Key performance metrics include sensitivity, sample preparation workflow, hydrolysis control, metabolite coverage, matrix compatibility, automation potential and throughput. GC–MS remains the workhorse for hair and sweat ultra-trace screening after derivatization. HPLC with UV, fluorescence or diode-array detection enables robust quantification of morphine and its glucuronides in pharmacokinetic and clinical settings. LC–MS facilitates the multiplexed analysis of DIM, its ester metabolites and its conjugates in a single, rapid run under gentle conditions to prevent ex vivo degradation. Recent advances such as high-resolution mass spectrometry and microsampling techniques offer new opportunities for sensitive and matrix-adapted analysis. By integrating validation parameters, forensic applicability, and evolving instrumentation, this review provides a practical roadmap for toxicologists and analysts navigating complex biological evidence. Full article

Background: Shancigu is a traditional Chinese medicine which is effective at clearing heat, detoxifying, dissipating masses, and resolving nodules. It consists of the dried pseudobulbs of orchids such as Cremastra appendiculata, or Pleione yunnanensis. To deeply understand the differences in the compositional and pharmacological active compounds in Shancigu, this study employed widely targeted metabolomics to analyze differential metabolites between two Shancigu species, C. appendiculata and P. yunnanensis. Methods: In this study, ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) was used to qualitatively, quantitatively, and differentially analyze the metabolites of C. appendiculata and P. yunnanensis. Results: Metabolite profiling identified 2890 compounds across 13 classes. Within these, 687 metabolites showed significant differential abundance (23.76% total), including 331 upregulated and 356 downregulated compounds. Pathway enrichment analysis revealed these differential metabolites primarily concentrated in stilbenoid biosynthesis (types I and II) and flavonoid aglycone biosynthesis. The most highly expressed metabolites in the Cremastra group were L-tyrosine, dopamine and 3,4-dihydroxybenzaldehyde-xylose-glucoside, while in the Pleione group, the most abundant metabolites were 3,5-dihydroxy-2’-methoxy-4-methylbibenzyl, Shancigusin F and aloifol I. C. appendiculata preferentially accumulates flavonoids and phenolic acids whereas P. yunnanensis favors terpenoid and nucleotide derivative production. Conclusions: This study identifies key differential metabolites in C. appendiculata and P. yunnanensis, providing basic data for the overall evaluation and breeding of Shancigu, laying a foundation for further quality control and precise medication of Shancigu. Full article

Heme metabolism is crucial for the function and longevity of legume nodules, with leghemoglobins (Lbs) serving as the primary heme sink and heme oxygenase (HO) mediating heme degradation. However, the precise functional relationship between HO and Lbs remains unclear. Here, we show that Lotus japonicus HO1 (LjHO1) is strongly induced in early-stage Lb-deficient nodules, but its expression gradually decreases during nodule development. Subcellular localization analysis revealed that LjHO1 is plastid-localized in uninfected cells of lb123 mutant nodules, consistent with its localization in wild-type nodules. Using CRISPR/Cas9, we generated a quadruple ho1lb123 mutant lacking LjHO1 and all three Lb isoforms in L. japonicus. Phenotypic analyses revealed that Lbs deficiency predominantly impairs nitrogen fixation, whereas loss of LjHO1 further reduces nodule formation. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis revealed that loss of Lbs strongly decreased heme accumulation, whereas LjHO1 deficiency slightly increased heme levels in nodules. These results demonstrate that Lbs are essential for heme accumulation and nitrogen fixation, while LjHO1 fine-tunes heme turnover, highlighting their complementary roles in maintaining nodule heme homeostasis and symbiotic efficiency. Full article

An integrative approach combining network pharmacology, molecular docking, and cellular assays was used to elucidate the potential mechanisms by which the n-butanol extract of Biebersteinia heterostemon ameliorates type 2 diabetes mellitus (T2DM). Chemical constituents of the n-butanol extract were identified via ultra-high-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry. Active compounds and T2DM-related targets were retrieved from public databases, and intersecting targets were identified. Protein–protein interaction (PPI) networks were constructed using the STRING database, while Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed via the DAVID database. A comprehensive “drug–compound–target–disease–pathway” network was established, and molecular docking was conducted to evaluate binding affinities of key compounds to core targets. Functional validation was performed in insulin-resistant cell models. Network pharmacology analysis identified 37 active constituents within the extract and 222 overlapping targets associated with T2DM. GO enrichment indicated involvement in protein phosphorylation, MAPK cascade activation, and negative regulation of apoptosis. Key signaling pathways included PI3K/AKT and lipid and atherosclerosis pathways. Molecular docking revealed strong binding affinities (binding energies ≤ −9.3 kcal·mol⁻¹) between core compounds—such as cheilanthifoline, glabridin, acetylcorynoline, skullcapflavone II, liquiritigenin, and dinatin—and pivotal targets including GAPDH, AKT1, TNF, SRC, EGFR, and PPARγ. In vitro experiments demonstrated that the extract significantly enhanced glucose uptake and glycogen synthesis in insulin-resistant cells, while suppressing oxidative stress and the expression of pro-inflammatory mediators such as TNF-α, MMP9, and IL-6. Collectively, B. heterostemon shows potential as an effective intervention for T2DM by targeting key molecular pathways, improving insulin sensitivity, and mitigating oxidative stress and inflammation in insulin-resistant cells. Full article

Objective: Hexafluoropropylene oxide dimer acid (HFPO-DA), also known as GenX, is widely used globally, raising concerns about its safety and public health implications. However, its toxicity mechanism remains unclear. The purpose of this study was to develop a reliable method for detecting HFPO-DA in mice and to investigate its absorption, distribution, and impact on hepatic lipid metabolism. Method: HFPO-DA levels were measured in the serum and eight tissues of C57BL/6J mice after oral administration using ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS). Lipid metabolites in the liver were also detected and analyzed. Results: HFPO-DA was rapidly absorbed into the bloodstream and widely distributed throughout all tested tissues. It penetrated the blood–brain barrier, with the highest concentration in the liver; however, long-term effects on the lungs also warrant attention. HFPO-DA disrupted liver lipid metabolism, leading to acylcarnitine accumulation while lowering triglycerides and cholesterol. Conclusion: This study on the pharmacokinetics and tissue distribution of HFPO-DA in mice following oral exposure revealed that HFPO-DA exacerbates liver injury by altering hepatic lipid metabolism. These findings provide theoretical support for toxicological studies on the emerging environmental pollutant HFPO-DA. Full article

The leaves of Apocynum venetum L. (A. venetum L.) are a functional food that plays an important role in antioxidation due to its high content of flavonoids and phenolic acids. Therefore, the extraction process of leaves of A. venetum L. is closely related to their activity. In this study, ultra-high-performance liquid chromatography (UHPLC) coupled with diode array detector (DAD), electrospray ionization (ESI), and quadrupole time-of-flight mass spectrometry (QTOF/MS) techniques has been established for qualitative and quantitative analysis of three phenolic acids and six flavonoids in the leaves of A. venetum L. Ultrasonic-assisted extraction conditions for the maximum recovery of phenolic and flavonoid compounds with a high antioxidation effect were optimized by response surface methodology (RSM). The optimum extraction conditions were as follows: ethanol concentration 64%, extraction time 20 min, and liquid-to-solid ratio 16:1 mL/g. The yields of three phenolic acids and six flavonoids under the optimal process were found to be 8.932 ± 0.091 and 20.530 ± 0.198 mg/g, respectively, which matched with those predicted (8.751 and 20.411 mg/g) within a 95% confidence level. Antioxidant activities based on ABTS and DPPH assays showed that the optimal extracts had strong activities compared with those of conventional reflux extraction methods. Moreover, the contribution of total and individual phenolic acids and flavonoids to antioxidant activity was also estimated by Pearson correlation analysis. Full article

Pistachio (Pistacia vera L.) holds significant importance due to its diverse applications and nutritional benefits. The nuts are rich in essential amino acids, antioxidants, fiber, healthy fats, and minerals, making them highly valuable for human nutrition. However, pistachios are significantly challenged by salinity stress, which negatively affects their growth and metabolism. Understanding the impact of salinity stress on pistachios is crucial for developing effective strategies to enhance their tolerance, improve growth, and ensure sustainable production in saline environments. To investigate the effects of salinity on energy metabolism and amino acid composition, we monitored key metabolites and free amino acid levels in UCB-1 pistachio leaves at 7- and 21-day salt stress treatments using Liquid Chromatography–Mass Spectrometry (LC-MS) and Ultra Performance Liquid Chromatography (UPLC). Our findings revealed that salinity affected nearly all analyzed metabolites, with varied patterns observed at different time points. Notably, all free amino acids except threonine accumulated significantly in response to salt stress. Meanwhile, reductions in 3PGA, Fru1,6bP, and Glu6P+Fru6P (glycolysis and Calvin cycle intermediates) suggest a decrease in photosynthetic activity, which may ultimately impact respiration rates. These results demonstrate that salinity stress affects both amino acid metabolism and central carbon metabolism, with the magnitude and pattern of these changes depending on the duration of exposure. The observed metabolic adjustments likely represent an adaptive response, enabling the plant to partially mitigate the detrimental effects of salt stress. Full article

Background/Objectives: Hypertension is a global health challenge. Zhengan Xifeng Decoction (ZXD), a classical traditional Chinese medicine, has shown clinical efficacy against hypertension. This study aimed to identify the bioactive constituents of ZXD and elucidate its antihypertensive mechanisms by integrating plasma UPLC–MS (ultra-performance liquid chromatography–mass spectrometry) analysis, network pharmacology, and molecular dynamics (MD) simulations. Methods: ZXD constituents and plasma-absorbed compounds were characterized by UPLC–MS. Putative targets (TCMSP, SwissTargetPrediction) were cross-referenced with hypertension targets (GeneCards, OMIM) and analyzed in a STRING protein–protein interaction network (Cytoscape) to define hub targets, followed by GO/KEGG enrichment. Selected protein–ligand complexes underwent docking, Prime MM-GBSA calculation, and MD validation. Results: A total of 72 absorbed components were identified, including 14 prototype compounds and 58 metabolites. Network pharmacology identified ten key bioactive compounds (e.g., liquiritigenin, isoliquiritigenin, and caffeic acid), 149 hypertension-related targets, and ten core targets such as SRC, PIK3CA, PIK3CB, EGFR, and IGF1R. Functional enrichment implicated cardiovascular, metabolic, and stress-response pathways in the antihypertensive effects of ZXD. Molecular docking demonstrated strong interactions between key compounds, including liquiritigenin, caffeic acid, and isoliquiritigenin, and core targets, supported by the MM-GBSA binding free energy estimation. Subsequent MD simulations confirmed the docking poses and validated the stability of the protein–ligand complexes over time. Conclusions: These findings provide mechanistic insights into the multi-component, multi-target, and multi-pathway therapeutic effects of ZXD, offering a scientific basis for its clinical use and potential guidance for future drug development in hypertension management. Full article

DNA direct reactive impurities (DDRIs) can react with nucleophilic sites of DNA, leading to mutations. The control strategies outlined in International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) M7 are based on the known compound structure of DDRIs. Non-target screening of DDRIs in drugs is still challenging due to the diversity of the species and the poor stability. In this study, a derivatization reagent including a reactive group and report group was designed to screen DDRIs. Based on the electrophilic theory of chemical carcinogenesis, an amine reagent was used as a reactive group to interact with DDRIs. Two derivatization reagents, p-methoxyaniline and p-methoxybenzoyl-β-alaninamide, were employed, each containing different chromatographic modification groups to mitigate matrix effects. The derivatization products were analyzed by ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry (UPLC-HRMS). Non-target screening for DDRIs was achieved by product ions filtering of the report group. Full article

The coloration of shellfish significantly influences both environmental adaptability and economic value. In the Jinjiang oyster (Crassostrea ariakensis), soft-body color varies between individuals, with an orange-yellow phenotype distinct from the milky white coloration of the wild type. To elucidate the compositional differences and molecular mechanisms underlying orange-yellow (designated as CaR) versus milky white (CaW) soft-body color in C. ariakensis, we conducted comparative ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) non-targeted and transcriptomic analyses. A total of 280 differential accumulation metabolites (DAMs) and 691 differentially expressed genes (DEGs) were detected between the CaR and CaW groups. The metabolite set enrichment analysis (MSEA) revealed that DAMs were significantly enriched in pigment metabolism pathways, including tyrosine metabolism, porphyrin metabolism, and lipid metabolism. Furthermore, genes associated with melanin synthesis and carotenoids conversions or transports were upregulated in the CaR vs. CaW group. These genes included Cyp4z1, Cyp4f22, Cyp17a1, Cyp1a5, Cyp2d28a, Lrp4, Aldh, and Tyr-3, potentially driving the accumulation of pheomelanin and carotenoids. This study demonstrates the vital roles of melanin and carotenoid metabolism in Jinjiang oyster body color formation, providing key insights into the molecular mechanisms of color determination in shellfish. Full article

This research provides a comprehensive evaluation of the phytochemical composition, antioxidant potential, and biological properties of four plant species with longstanding use in ethnobotanical traditions: Calendula officinalis, Mentha × piperita, Urtica dioica, and Juglans regia. Plant extracts were obtained using a range of solvent systems and subsequently analyzed for their content of total polyphenols, flavonoids, and phenolic acids. Ultra-high-performance liquid chromatography coupled with mass spectrometry (UHPLC-MS) enabled the accurate identification and quantification of major polyphenolic constituents. The antioxidant capacity was assessed through a series of in vitro assays, and elemental analysis was conducted to determine microelement content. To evaluate potential ecological implications, acute toxicity was tested using Daphnia magna, while phytotoxic effects were also examined. The results demonstrate pronounced antioxidant activity along with notable biocidal and soil-enhancing properties. These findings underscore the potential of such plant-based formulations as sustainable alternatives to conventional agrochemicals and highlight the relevance of integrating traditional botanical knowledge with modern strategies for enhancing soil quality, crop performance, and environmental sustainability. Full article

Background/Objectives: Bangkeehwangkee-tang (BHT) is a traditional herbal formula composed of six medicinal herbs: Sinomenii Caulis et Rhizoma, Astragali Radix, Atractylodis Rhizoma Alba, Zingiberis Rhizoma Recens, Zizyphi Fructus, and Glycyrrhizae Radix et Rhizoma. BHT has been widely used for its immunomodulatory and anti-inflammatory effects. This study aimed to develop a reliable analytical method for the simultaneous determination of 22 marker compounds to ensure consistent quality control and to ensure consistent efficacy in both clinical and non-clinical studies of BHT. Methods: An ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method based on multiple reaction monitoring was developed and validated for the simultaneous determination of 22 marker compounds in BHT. The method was evaluated for selectivity, linearity (coefficient of determination, r²), sensitivity (limit of detection (LOD) and limit of quantification (LOQ)), accuracy (recovery), and precision (relative standard deviation (RSD)) in accordance with guidelines. Results: The developed method exhibited excellent selectivity and linearity (r² ≥ 0.9913) for all target compounds. The LOD and LOQ ranged from 0.09 μg/L to 326.58 μg/L and 0.28 μg/L to 979.75 μg/L, respectively. The recovery ranged from 90.36% to 113.74%, and precision (RSD) was ≤15%, confirming the method’s reliability. The application of the method to various BHT samples revealed substantial variations in the marker compound contents, particularly for sinomenine, magnoflorine, and glycyrrhizin. Conclusions: These findings highlight the necessity for standardized quality control of BHT and demonstrate that the developed UPLC–MS/MS method is a practical and reliable tool for performing quality assessment of traditional herbal formulas. Full article

Numerous factors, including genetic, environmental, and nutritional, are involved in testicular development and spermatogenesis. However, little is known about the effects of seasonal factors on pre-sexual maturity testicular development in Hu rams, which are famous for their high fertility and year-round estrus onset. This study explored the effect of the birth season on testicular development and spermatogenesis in Hu sheep. Thirty-six 6-month-old male lambs born in summer (n = 18) and winter (n = 18) were selected for analysis. Results showed that summer-born lambs exhibited significantly higher cauda sperm density (102.65 ± 9.56 vs. 16.86 ± 2.02 × 10⁷/g), antioxidant indices such as superoxide dismutase (SOD: 6.29 ± 1.01 vs. 4.09 ± 0.25 U/mgprot), and higher expression levels of glutathione peroxidase 3 (GPX3), glutathione peroxidase 4 (GPX4), and copper/zinc superoxide dismutase (Cu/Zn-SOD) than winter-born lambs. Conversely, the malondialdehyde content (1.08 ± 0.32 vs. 2.13 ± 0.34 nmol/mgprot) was significantly lower in the summer-born group (p < 0.05) than in the winter-born group. A total of 44 differential oxylipins and 326 differentially expressed genes (DEGs) were screened by ultra-performance liquid chromatography–tandem mass spectrometry and transcriptomics, respectively. An integrated analysis of oxylipins and transcriptomics revealed that these differential molecules were enriched in metabolic pathways. Notably, downregulated DEGs (e.g., UAP1L1 and NAT8L) were significantly correlated with upregulated differential oxylipins (e.g., epoxyeicosatrienoic acids and dihydroxyeicosatrienoic acids). These results indicate that compared to the winter-born group, the testicular tissues of summer-born rams showed stronger testicular antioxidant capacity and lower lipid peroxidation at the sexual maturity stage, which contributes to spermatogenesis. Full article

Background/Objectives: Watermelon (Citrullus lanatus) is a natural dietary source of L-citrulline and L-arginine, the two amino acids involved in nitric oxide (NO) production and vasodilation. Pre-clinical and clinical studies using isolated amino acids or watermelon extracts suggest blood pressure (BP)-lowering potential; however, limited research has been conducted on the impact of watermelon flesh (WM) on BP in adults at risk for hypertension. Therefore, the primary objective of this study was to assess the effect of daily WM intake for four weeks on 24 h ambulatory BP in adults with elevated blood pressure. The secondary outcomes of this study include changes in glucose and insulin markers, lipid profile, NO, L-citrulline, L-arginine, asymmetric dimethylarginine (ADMA) concentrations, and the L-arginine/ADMA ratio. Methods: In this randomized, placebo controlled parallel study design, 39 adults (age: 41 ± 14 years, BMI: 31 ± 6 kg/m², mean ± SD) with elevated BP were randomly assigned to one of three groups for a 4-week intervention: control (0 g WM), WM-1 cup (152 g/day), or WM-2 cups (304 g/day). Ambulatory BP was measured over 24 h at baseline and the end of the intervention period. Fasting plasma samples were analyzed for metabolic biomarkers on a clinical analyzer and NO using a colorimetric assay. L-citrulline, L-arginine, and ADMA were analyzed using an ultra-high-performance liquid chromatography triple quadrupole mass spectrometer (UHPLC-QQQ-MS/MS). Statistical analyses were conducted using SPSS software (IBM SPSS Statistics, Version 29.0.0). Results: After 4 weeks, mean 24 h ambulatory BP was 130.2 ± 3.9 mm Hg (control), 130 ± 3.2 mm Hg (WM-1 cup), and 124.9 ± 3.9 mm Hg (WM-2 cups), with no statistically significant differences between study interventions (p > 0.05). Similarly, no significant changes were observed in fasting plasma glucose, insulin, lipid profile, or NO concentrations. However, plasma L-arginine concentrations and L-arginine/ADMA ratios significantly increased in the WM groups compared to the control (p = 0.009) after adjusting for age, BMI, race, and gender in the statistical model. Conclusion: Overall, BP was not significantly different after two different doses of watermelon compared to control; however, improvements in NO synthesis pathway precursors (L-arginine, ADMA) suggest potential for dietary modulation to support endothelial function and BP regulation. Full article