Search Results (168)

Background: Catalpa bungei ‘Jinsi’ has excellent wood properties and golden texture, which is widely used in producing furniture and crafts. The lignin content and structural composition often determine the use and value of wood. Hence, investigating the characteristics of the annual dynamics of lignin anabolic metabolites in C. bungei ‘Jinsi’ and analyzing their synthesis pathways are particularly important. Methods: We carried out targeted metabolomics analysis of lignin synthesis metabolites using ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) on the xylem samples of C. bungei ‘Jinsi’ in February, April, July, October 2022, and January 2023. Results: A total of 10 lignin synthesis–related metabolites were detected: L-phenylalanine, cinnamic acid, p-coumaraldehyde, sinapic acid, p-coumaric acid, coniferaldehyde, ferulic acid, sinapaldehyde, caffeic acid, and sinapyl alcohol (annual total content from high to low). These metabolites were mainly annotated to the synthesis of secondary metabolites and phenylpropane biosynthesis. The annual total content of the 10 metabolites showed the tendency of “decreasing, then increasing, and then decreasing”. Conclusions: C. bungei ‘Jinsi’ is a typical G/S-lignin tree species, and the synthesis of G-lignin occurs earlier than that of S-lignin. The total metabolite content decreased rapidly, and the lignin anabolism process was active from April to July; the metabolites were accumulated, and the lignin anabolism process slowed down from July to October; the total metabolite content remained basically unchanged, and lignin synthesis slowed down or stagnated from October to January of the following year. This reveals the annual dynamic pattern of lignin biosynthesis, which contributes to improving the wood quality and yield of C. bungei ‘Jinsi’ and provides a theoretical basis for its targeted breeding. Full article

A sensitive method was developed for detecting diazepam residues in aquatic products using magnetic dispersive solid-phase extraction (MDSPE) coupled with ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS). Samples extracted with 1% ammonia–acetonitrile were purified using synthesized Fe₃O₄@SiO₂-PSA nanoparticles via MDSPE before UPLC-MS/MS analysis. Separation was performed on a C₁₈ column with gradient elution using 0.1% formic acid–2 mM ammonium acetate/methanol. Detection employed positive electrospray ionization (ESI⁺) in multiple reaction monitoring (MRM) mode. Characterization confirmed Fe₃O₄@SiO₂-PSA’s mesoporous structure with excellent adsorption capacity and magnetic properties. The method showed good linearity (0.1–10 μg/L, r > 0.99) with an LOD and LOQ of 0.20 μg/kg and 0.50 μg/kg, respectively. Recoveries at 0.5–15.0 µg/kg spiking levels were 74.9–109% (RSDs 1.24–11.6%). This approach provides rapid, accurate, and high-precision analysis of diazepam in aquatic products, meeting regulatory requirements. Full article

Lolium perenne (Poaceae), a perennial forage, has high economic and nutritional value. It is often used as a replacement control for some invasive plants, as it has achieved good ecological and economic effects. However, its control effects, allelochemicals, allelopathic effects, release pathways, and contents are still unclear in the process of L. perenne replacement control of an invasive plant, Ageratina adenophora (Asteraceae). Therefore, it is necessary to reveal the mechanism of L. perenne replacement control of A. adenophora from the perspective of allelopathy. In this study, L. perenne could effectively inhibit the growth of A. adenophora in the competition assay. In addition, seven norsesquiterpenes (1–7) were isolated and identified from the whole plant of L. perenne, and most of the compounds exhibited potent allelopathic effects on the growth of A. adenophora and one model plant (Lactuca sativa, Asteraceae). Moreover, some active compounds were released into the environment through root secretion and rainwater leaching, and their contents were determined by UPLC-MS/MS (Ultra Performance Liquid Chromatography Tandem Mass Spectrometry). Our results elucidated the allelopathic mechanism of L. perenne’s replacement control, A. adenophora, and provided a theoretical basis for the development of norsesquiterpenes from L. perenne. Full article

Background/Objectives: Endophytes can produce bioactive metabolites similar to their host plants. CM-YJ44 (Pseudomonas protegens CHA0, 99.24% similarity), an endophyte from Dendrobium officinale, has not yet validated hypoglycemic potential. This study aimed to evaluate its anti-insulin resistance (IR) activity and metabolite profile. Methods: The fermentation broth of CM-YJ44 was separated into three fractions (CM-YJ44-1, -2, and -3) using semi-preparative high-performance liquid chromatography (pre-HPLC). An IR HepG2 cell model was constructed to evaluate their glucose uptake capacity. CM-YJ44-3 was further tested for oxidative stress, inflammatory, and insulin signaling pathway activation. Metabolites in CM-YJ44-3 were preliminarily identified using the Q Exactive Focus LC-MS system (QE), and the dendrobine content was quantified by ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS). Molecular docking was performed to predict the binding affinities between dendrobine and target proteins. Results: Among the three fractions, CM-YJ44-3 significantly reduced nitric oxide (NO) and reactive oxygen species (ROS) levels in IR cells, enhanced glycogen synthesis, upregulated the activities of pyruvate kinase (PK) and hexokinase (HK), and suppressed the expression of inflammatory factors. Its mechanism of action was mainly through activation of the IRS1/PI3K/Akt/GSK3β/GLUT4 signaling pathway. QE analysis preliminarily identified 24 metabolites in CM-YJ44-3. Quantitative analysis by UPLC-MS/MS showed that the dendrobine content was 78.73 ± 4.29 ng/mL. Molecular docking results indicated that dendrobine exhibited binding energies below −5 kcal/mol with multiple target proteins involved in this signaling pathway, suggesting it may be a key bioactive component responsible for the anti-IR effect. Conclusions: This study provides the first evidence of hypoglycemic bioactive metabolite production by strain CM-YJ44, indicating its potential as a novel microbial candidate for alleviating IR. Full article

Background/Objectives: Saussurea obvallata (DC.) Edgew., Asteraceae, is a traditional medicinal herbnative to the Qinghai–Tibet Plateau (QTP). Pharmacological investigationshave validated its pharmacological effects in anti-tumor, anti-inflammatory, heat-clearing, detoxifying, and analgesia. S. obv is presently facing habitat fragmentation and population decline. Therefore, we analyzed its genetic and chemical diversity to provide a scientific basis for the conservation and sustainable use of S. obv. Methods: Seven populations of S. obv were sampled from Xizang, China. The genetic diversity was analyzed using inter-simple sequence repeat (ISSR) markers, and metabolites were identified by ultra-high-performance liquid chromatography-tandem-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS). Correlation analysis among genetic diversity, differential metabolites, and climatic factors were performed by R. Results: The genetic diversity among and within populations were both lowly and significantly correlated with geographical distance, showing a decreasing trend from east to west of the QTP. A total of 110 compounds were identified, including flavonoids, phenylpropanoids, lipids, fatty acids, terpenoids, alkaloids, etc. The metabolite contents among populations varied greatly and were related to environmental factors, mainly annual mean temperature and temperature fluctuation. The genetic diversity had little effect on the metabolic differences. Conclusions: These findings provided valuable baseline information for the conservation and pharmacological utilization of S. obv. Meanwhile, further research is necessary for the efficacy evaluation of anti-inflammatory, anti-tumor, radiation protection, and scar removal both in vitro and in vivo. Full article

Amanita species are widely distributed worldwide. Many of these species are poisonous and can cause health problems, resulting in morbidity and mortality. The toxins responsible for poisoning are amatoxins, aminohexadienoic acid, ibotenic acid, muscimol and muscarines, which damage the liver, kidney, central nervous system and parasympathetic nervous system. In recent years, several toxins have been discovered from different poisonous mushrooms. In this study, multiwalled carbon nanotube purification and ultrahigh-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) was used for the sensitive detection and targeted quantitative screening of 12 mushroom toxins (muscarine, two isoxazole derivatives, three tryptamine alkaloids, three amatoxins and three phallotoxins) from Amanita citrina, A. citrina var. grisea and A. sinocitrina. This study found that buiotenine, one of the tryptamine alkaloids, was detected in A. citrina and A. sinocitrina with an average content of 2.90 and 1.19–6.70 g/kg (n = 3) in the dried mushrooms, respectively. None of the 12 common toxins were discovered in A. citrina var. grisea. These results provide reference data for future research on the role of toxins in the evolution of Amanita mushrooms. Future studies should explore the biosynthetic pathways and ecological roles of these toxins in Amanita species. Full article

A high-throughput method for the determination of a variety of chemical hazards in poultry muscle and egg samples was established via ultra-performance liquid chromatography–tandem triple quadrupole mass spectrometry (UPLC–QqQ-MS). The sample preparation procedure was developed based on this quick, easy, cheap, effective, rugged, and safe (QuEChERS) method and validated for 280 chemical hazards potentially present in poultry products. The target compounds in poultry samples were extracted with a 1% formic acid–acetonitrile solution (15:85, v/v), and the metal ions in the matrix were chelated by adding ethylenediaminetetraacetic acid disodium salt (Na₂EDTA). The supernatant was purified using Enhanced Matrix Removal (EMR) lipid sorbent. Chromatographic gradient separation was performed on an ACQUITY UPLC BEH C18 (2.1 mm × 100 mm, 1.7 μm) column with multiple reaction monitoring (MRM) under both negative- and positive-ion mode. Internal standard calibration or matrix-matched calibration was used for the quantitation. The results showed that good linearity was achieved for each target compound with correlation coefficients (R²) ≥ 0.99. The limits of detection (LODs) ranged from 0.05 to 10 µg/kg, and the acceptable limits of quantification (LOQs) were determined to be 0.1–20 µg/kg for all 280 compounds. Approximately 90% of the target compounds exhibited mean recoveries ranging from 60% to 120%, with relative standard deviations (RSDs) within 16.2%. This method can be used for the high-throughput rapid detection of prohibited drug residues in poultry eggs due to its easy operation and high accuracy. It was applied in real sample detection, and 43 chemicals including metronidazole were found in 211 poultry samples, with a concentration range of 0.11–638 μg/kg. Full article

Exposure to patulin (PAT) poses a significant health risk to animals, emphasizing the need for natural, safe substances to mitigate toxicity. Acremonium terricola culture (ATC), a fungal fermentation-derived feed additive, is known for its antioxidant properties, yet its potential to alleviate mycotoxin-induced toxicity remains largely uninvestigated. In this study, the ethanol extracts from the ATC (EEAT) were prepared with a total phenolic content of 67.9 mg GAE/g and a total flavonoid content of 32.7 mg RE/g. Ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UPLC-ESI-MS/MS) analysis was employed to investigate the bioactive components in EEAT. In PAT-exposed Caenorhabditis elegans models, EEAT treatment significantly enhanced locomotory capacity and elevated antioxidant enzyme activities by 63.1% (SOD) and 90.1% (GSH-ST), respectively. Molecular docking analysis revealed that key active compounds in EEAT, such as coumarin, succinic acid, and trigonelline, exhibited effective binding affinities to potential targets SIR-2.1 and DAF-2. Notably, coumarin and trigonelline were most effective in alleviating PAT toxicity, as evidenced by rescued locomotor rates and oxidative impairment in C. elegans. Our findings not only elucidate the molecular basis of EEAT-mediated PAT mitigation but also establish A. terricola culture as a sustainable antioxidant. Full article

This study established a dual analytical workflow integrating thermal desorption–electrospray ionization–tandem mass spectrometry (TD-ESI-MS/MS) for rapid qualitative screening and ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) for confirmatory quantification of 17 psychoactive substances and metabolites across six classes (opioids, amphetamine-type stimulants, cocaine, ketamine-type drugs, cannabinoids, and etomidate analogs) in hair matrices. Validation of the TD-ESI-MS/MS method demonstrated its sensitivity (limits of detection: 0.1–0.2 ng/mg) and precision (<19.3%), with matrix effects controlled to <19.6%. The TD-ESI-MS/MS method achieved an analysis time of 1 min per sample, enabling high-throughput screening with a sensitivity >85.7% and a specificity >89.7% for the 17 analytes. UPLC-MS/MS confirmation validated the screening results with accuracy rates of 89.7–99.8%. An analysis of specimens confirmed positive identified etomidate analogs as the predominant psychoactive substances (73.6%), with a lower prevalence of amphetamine-type stimulants (12.5%), ketamine-type drugs (9.0%), and opioids (2.8%). The polydrug use patterns identified concurrent etomidate–amphetamine consumption (n = 5) and complex analog combinations (etomidate–isopropoxate–metomidate, n = 13), suggesting evolving abuse trends. Despite limitations in the temporal resolution and representativeness of the cohort, this study demonstrated the viability of TD-ESI-MS/MS for bridging forensic and public health priorities. Future work should focus on optimizing the durability of the ion source for TD-ESI and validating this method across diverse populations to enhance its generalizability. Full article

Rice is sensitive to high temperatures at the seedling stage. In the present study, a combined analysis of transcriptome and metabolome was performed on a heat-resistant accession, DY80, from Dongxiang wild rice and a heat-sensitive variety, R974, under heat stress at the seedling stage. The results of the transcriptome and metabolome analyses were verified through qRT-PCR and ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) analysis. We found that there were 1817 and 561 differentially expressed genes (DEGs) unique in DY80 and R974 under heat stress, respectively. The elite genes for the heat stress involved in Dongxiang wild rice may include upregulated genes in the pathway of unfolded protein binding; downregulated genes in the pathways of chlorophyll biosynthetic process, and cysteine and methionine metabolism; and photosystem I, photosystem II, and unchanged genes in the pathways of the anchored component of the plasma membrane, cell wall biogenesis, and photosynthesis-antenna proteins. Moreover, a total of 301 and 28 metabolites were identified as unique in DY80 and R974 after heat treatment, respectively. Further analyses showed that malic acid, stearic acid, and L-threonine might be causal metabolites, contributing to strong heat resistance in Dongxiang wild rice. These findings provide new insights into the mechanisms of heat resistance in rice. Full article

Heterocyclic aromatic amines (HAAs), known for their mutagenic and carcinogenic potential, are formed during the heating of protein-rich food items. Detecting HAAs swiftly and accurately poses challenges due to complex food matrices and low HAA concentrations. In this study, a simple and efficient magnetic solid-phase extraction (MSPE) strategy was developed for the simultaneous isolation and enrichment of three HAAs such as 2-amino-3,4,8-trimethylimidazo [4,5-f]quinoxaline (4,8-DiMeIQx), 2-amino-3,8-dimethylimidazo [4,5-f]quinoxaline (MeIQx), and 2-amino-3-methylimidazo [4,5-f]quinoline (IQ) in processed meats, employing the magnetic covalent organic framework Fe₃O₄@MOF-545-AMSA as an adsorbent. It was synthesized via a solvothermal method, with Fe₃O₄ as the magnetic core. Its building blocks are as follows: zirconium (Zr) as the coordination metal ion, tetrakis(4-carboxyphenyl)porphyrin and benzoic acid as organic ligands, and aminomethanesulfonic acid (AMSA). This composite captures targeted HAAs efficiently by exploiting the unique porous MOF-545-AMSA structure, specific metal–ligand coordination, and AMSA’s amino and sulfonic acid groups. The quantification of HAAs was achieved through the combination of Ultra-Performance Liquid Chromatography–Tandem Mass Spectrometry (UPLC-MS/MS) and MSPE, demonstrating satisfactory linearity (R² ≥ 0.9917), high recovery rates (83.7–111.0%), and low detection limits (0.1–1.0 μg/kg). Moreover, an automated high-throughput detection system was developed using MSPE to assess the presence of HAAs in meat products. Full article

Gardeniae Fructus (GF), the desiccative mature fruitage of Gardenia jasminoides J. Ellis (G. jasminoides), is a traditional herbal medicine in China with potential value against skin photodamage. However, the phytochemical basis and mechanisms underlying GF’s anti-photodamage effects remain unclear. In this study, the chemical components in GF extract (GFE) were analyzed using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS), and iridoids were identified as the main components. The antioxidant, anti-inflammatory, and barrier-repair effects of GFE in UVB-induced photodamage were assessed through in vitro experiments. Additionally, the potential mechanisms of GFE against skin photodamage were predicted using proteomics and network pharmacology. The results showed that GFE significantly increased the levels of total superoxide dismutase (T-SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) induced by UVB, while decreasing reactive oxygen species (ROS) and malondialdehyde (MDA) contents. GFE also inhibited the secretion of interleukin-6 (IL-6) and interleukin-1β (IL-1β). Additionally, GFE upregulated the expression of filaggrin (FLG), loricrin (LOR), and involucrin (IVL) in 3D epidermal skin models. Proteomic analysis and network pharmacology indicated that the iridoid components identified in GFE ameliorated UVB-induced damage probably by regulating cell cycle-related proteins and signaling pathways, though this requires further experimental confirmation. Overall, the results provide essential evidence to support the development of GFE as a skincare active ingredient. Full article

Sample pretreatment is one of the most important steps in guaranteeing the success of a chromatographic analysis. The selected methodology must ensure simultaneously that a sample is “clean” enough for analysis and that the target analytes are not removed in the process. This can be especially difficult when working with complex matrices such as natural waters and wastewater. For pharmaceutical active compounds (PhACs) analysis by solid-phase extraction (SPE) followed by ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS), and due to the high level of organic matter in wastewater, the water samples are filtered consecutively through three filters, a paper filter, a glass microfiber filter of 1 µm, and a Nylon filter of 0.45 µm. This filtration allows the sample’s passage through the SPE cartridge to be faster, and there is no cartridge clogging, allowing for greater efficiency in the adsorption process. The big question is whether the PhACs are eliminated during filtration, since they may be adsorbed to organic matter. This work aimed to determine if the best approach for quantifying PhACs in wastewater and surface waters would be to filter them prior or to perform SPE directly. Both approaches analyzed a total of 26 PhACs. Turbidity (TUR) and permanganate index (PI) were determined, and their values were high for samples with a high organic matter content. A statistical analysis was performed to determine the best approach to treat these water samples and whether any correlation existed between PhAC concentrations, PI, and TUR. The PhAC quantification shows a positive correlation with TUR and a negative correlation with PI for most of the target PhACs. However, there are not significantly different results for filtered and not-filtered wastewater samples. Full article

Dinotefuran and clothianidin belong to the third generation of nicotinic insecticides and are widely used in crop pest control. It is necessary to detect their residues in food. The time-resolved fluorescent microspheres lateral flow immunoassay (TRFMs-LFIA) has the advantages of high sensitivity, short duration, and simple operation and is suitable for rapid field testing. In this study, two haptens (FCA-1, FCA-2) were synthesized in three steps and conjugated to the carrier proteins to obtain artificial antigens, which were subsequently used for monoclonal antibody preparation. A TRFMs-LFIA based on monoclonal antibodies was established to detect dinotefuran and clothianidin residues in food. The limit of detection (LOD) for dinotefuran was 0.045 ng/mL, with an IC₅₀ of 0.61 ng/mL and a linear range (IC₂₀~IC₈₀) of 0.12~3.11 ng/mL. The LOD for clothianidin was 0.11 ng/mL, with an IC₅₀ of 0.94 ng/mL and a linear range (IC₂₀~IC₈₀) of 0.24~3.65 ng/mL. Cross-reactivity rates with seven tested structural analogs were less than 1.5%. The pretreatment method was optimized for wheat, cucumber, and cabbage samples, which was time-saving (20 min) and easy to operate. The average recovery rates ranged from 88.0% to 114.8%, with the corresponding coefficients of variation appearing (CV) between 1.9% and 13.5%. The results of actual wheat, cucumber, and cabbage samples detected by the established TRFMs-LFIA were consistent with those of Ultra-Performance Liquid Chromatography coupled with Tandem Mass Spectrometry (UPLC-MS/MS). These results demonstrate that the established TRFMs-LFIA is sensitive, accurate, rapid, and suitable for real sample detection. Full article

Agrimonia pilosa Ledeb. (APL), a traditional Chinese herb frequently employed by Professor Zhou Zhongying, a master of traditional Chinese medicine, for colorectal cancer treatment, is rich in polyphenols with potential anti-tumor properties. To elucidate its bioactive components, this study developed a two-step purification process combining macroporous resin adsorption and liquid–liquid extraction to enrich polyphenols from APL (APLs). The adsorption/desorption mechanisms of APLs on macroporous resins were systematically investigated through resin screening, adsorption kinetics, and thermodynamics. The Langmuir isotherm model confirmed the adsorption process as spontaneous and exothermic. Pseudo-second-order kinetics effectively described the adsorption behavior of D101 resin. Optimized adsorption and column elution parameters were established, followed by liquid–liquid extraction for further purification. The components were compared and analyzed by ultra-performance liquid chromatography and quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-Zeno-TOF-MS/MS). It was preliminarily identified that 29 polyphenols were mainly concentrated in water-saturated n-butanol (BEA) and ethyl acetate (ECA) extract fractions. Quantitative analysis using ultra-high performance liquid chromatography–triple quadrupole liquid chromatography–mass spectrometry (UHPLC-C-QTRAP-MS/MS) revealed higher contents of catechin (66.67 ± 1.33 ng·mg⁻¹), hyperoside (382.56 ± 3.65 ng·mg⁻¹), and chlorogenic acid (10.60 ± 0.05 ng·mg⁻¹) in BEA compared to ECA (46.00 ± 2.00, 239.40 ± 2.60, and 3.42 ± 0.01 ng·mg⁻¹, respectively). In vitro experiments demonstrated that BEA exhibited superior antiproliferative activity (IC₅₀: 434.5 μg·mL⁻¹) and significantly inhibited CT26 tumor cell migration compared to ECA (IC₅₀: 672.5 μg·mL⁻¹). The enhanced biological activity of BEA may be due to its higher polyphenol content, suggesting that these compounds mediate their anti-tumor effects through different biochemical pathways. This work lays the foundation for exploring the multi-target mechanism of anti-tumor effects of APLs. Full article