Ultra-high Performance Liquid Chromatography Research Articles

A Comprehensive Investigation of Lipid Profile During the Solid-State Fermentation of Rice by Monascus purpureus

Red yeast rice is a nutraceutical fermented product used worldwide for the symptomatic relief of dyslipidemia and cardiovascular disease. However, the fermentation-induced lipid transformation from rice to red yeast rice remains unclear. Herein, an ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry method was developed for the comprehensive lipid analysis during fermentation. A total of 246 lipids fall in 21 subclasses were annotated in rice and red yeast rice, including 37 lysophospholipids, 14 phospholipids, 29 diglycerides, 114 triglycerides and fatty acid (15 species), ceramide (12 species), hexosylceramide (3 species), sitosterol ester (2 species), monogalactosyldiacylglycerol (2 species), digalactosyldiacylglycerol (2 species), monogalactosylmonoacylglycerol (8 species), digalactosylmonoacylglycerol (5 species), coenzyme Q (1 species), acyl hexosyl campesterol ester (1 species), and acylcarnitine (1 species). Results showed that lipid profiles changed, and new lipid species emerged. Notably, 18 medium- and long-chain triacylglycerols and triacylglycerols with short-chains were tentatively identified. These triacylglycerols also show the effects of body fat accumulation reduction, and hypolipidemic and hypoglycemic activities. Furthermore, lipid species that were profoundly changed were quantified, and the dynamic changes were investigated. This study clarified the molecular species and compositional changes in fermented rice from lipid aspect.

Protocol for the Determination of Total Iodine in Iodized Table Salts Using Ultra-High-Performance Liquid Chromatography

Potassium iodate and potassium iodide are commonly fortified in iodized table salt, which must be continuously monitored to maintain quality. Our study reported an optimized detection method for total iodine in iodized table salt using 0.5 M sodium bisulfite as the reducing agent. The iodized table salt (0.5 g) was dissolved in 0.5 M sodium bisulfite solution prior to injection in ultra-high-performance liquid chromatography (UHPLC) coupled with a diode array detector using a weak anion-exchange column (2.1 mm × 150 mm, 5 μm). Iodide was eluted at 9.92 ± 0.06 min (λ = 223 nm) when an isocratic mobile phase of 1:1 (v/v) methanol/120 mM phosphate buffer mixed with tetrasodium pyrophosphate (pH 3.0) was running at 0.20 mL/min (15 min). Iodide was detected as total iodine from 10.0 to 50.0 mg/kg with a limit of detection (LOD) of 1.2 mg/kg and a limit of quantification (LOQ) of 3.7 mg/kg. The method was validated with relative standard deviations (RSDs) of 4.2%, 0.4%, 1.6%, and 0.8% for accuracy, repeatability, intermediate precision, and robustness, respectively. The determination of total iodine was successful on six (6) samples (n = 3), which recovered 87.2–106.9% of iodate and iodide spike. Thus, this study provides a validated protocol for the determination of total iodine in iodized table salt using 0.5 M sodium bisulfite.

Metabolome profiling dissects the oat (Avena sativa L.) innate immune response to Pseudomonas syringae pathovars.

One of the most important characteristics of successful plant defence is the ability to rapidly identify potential threats in the surrounding environment. Plants rely on the perception of microbe-derived molecular pattern chemicals for this recognition, which initiates a number of induced defence reactions that ultimately increase plant resistance. The metabolome acts as a metabolic fingerprint of the biochemical activities of a biological system under particular conditions, and therefore provides a functional readout of the cellular mechanisms involved. Untargeted metabolomics was applied to decipher the biochemical processes related to defence responses of oat plants inoculated with pathovars of Pseudomonas syringae (pathogenic and non-pathogenic on oat) and thereby identify signatory markers that are involved in host or nonhost defence responses. The strains were P. syringae pv. coronafaciens (Ps-c), P. syringae pv. tabaci, P. syringae pv. tomato DC3000 and the hrcC mutant of DC3000. At the seedling growth stage, metabolic alterations in the Dunnart oat cultivar (tolerant to Ps-c) in response to inoculation with the respective P. syringae pathovars were examined following perception and response assays. Following inoculation, plants were monitored for symptom development and harvested at 2-, 4- and 6 d.p.i. Methanolic leaf extracts were analysed by ultra-high-performance liquid chromatography (UHPLC) connected to high-definition mass spectrometry. Chemometric modelling and multivariate statistical analysis indicated time-related metabolic reconfigurations that point to host and nonhost interactions in response to bacterial inoculation/infection. Metabolic profiles derived from further multivariate data analyses revealed a range of metabolite classes involved in the respective defence responses, including fatty acids, amino acids, phenolic acids and phenolic amides, flavonoids, saponins, and alkaloids. The findings in this study allowed the elucidation of metabolic changes involved in oat defence responses to a range of pathovars of P. syringae and ultimately contribute to a more comprehensive view of the oat plant metabolism under biotic stress during host vs nonhost interactions.

Unveiling potential functional applications of grape pomace extracts based on their phenolic profiling, bioactivities, and circular bioeconomy

Abstract Grape pomace (GP), a by-residue from the wine industry, contains bioactive molecules such as phenolic compounds, and anthocyanins, among others, with potential health benefits. In the current study, these bioactive molecules were extracted from GP of different Vitis vinifera L. varieties (Tinta Negra, Complexa, Malvasia Roxa, Malvasia, Sercial, Verdelho, Boal, Terrantez) using the micro quick, easy, cheap, effective, rugged, and safe (µQuEChERS) procedure. The GP extracts were investigated using ultra-high performance liquid chromatography (UHPLC-PDA) to establish the phenolic fingerprint, and by in vitro assays to assess the antibacterial, anti-inflammatory, and antioxidant activities. Nine phenolic compounds were identified and quantified in GP extracts, with gallic acid (ranging from 10.4 to 12.9 g/100 g), catechin (2.97 to 5.08 g/100 g), quercetin (2.17 to 2.85 g/100 g), and trans-resveratrol (0.28 to 1.82 g/100 g) being the most prominent. GP from the Complexa variety exhibited the highest levels of total anthocyanin content (TAC, 6.67 mgCGE/100 g), total phenolic compounds (TPC, 4727 mgGAE/100 g), and antioxidant activity (DPPH, 9472 mgTE/100 g), while the Tinta Negra variety had the highest total catechin content (TCC, 947 mgCATE/100 g). A strong correlation (p < 0.001) was observed between the TPC-TAC, TPC-DPPH, DPPH-TAC, and TAC-TCC. Moreover, o-coumaric acid and quercetin are significantly (p < 0.001) correlated with TPC, TAC, TCC, and DPPH assays. The investigated GP extracts, at a concentration of 100 µg/mL, showed promising inhibition of albumin protein denaturation compared to aspirin (reference standard). The findings showed that the GP extracts were more useful at inhibiting Staphylococcus aureus compared to Escherichia coli. It is important to emphasise that the GP extracts demonstrated antioxidant, anti-inflammatory, and antibacterial properties, positioning it as an agro-waste with promising potential for use in the development of innovative functional foods, dietary supplements, and cosmetics, aligning with the circular bioeconomy model for its valorisation.

Sini decoction alleviates LPS-induced sepsis partly via restoration of metabolic impairments in the hypothalamic-pituitary-adrenal microenvironment.

The hypothalamic-pituitary-adrenal (HPA) axis plays a vital role in the protection against sepsis. Sini decoction (SND) could improve HPA axis function. This work aimed to explore the effective mechanism of SND against lipopolysaccharide (LPS)-induced sepsis in rats from the metabolic regulation of the HPA axis microenvironment. We evaluated the multiorgan injury-associated enzymatic indicators and histopathological changes as well as the ultrastructural changes in the hypothalamus, pituitary gland, and adrenal gland associated with LPS-induced sepsis. Serum inflammatory cytokines, corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and corticosterone (CORT) were determined by ELISA. The target tissues metabolomics of the HPA axis (hypothalamus, pituitary gland, and adrenal gland), based on ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOFMS), were conducted to dissect the metabolic network regulated by SND. Western blotting was further used to validate the key metabolic pathways. In addition, the absorbed chemical constituents in serum and cerebrospinal fluid were identified by UHPLC-Q-TOFMS combined with solid-phase extraction. Forty and twenty-three components of SND were absorbed into the serum and cerebrospinal fluid, respectively. SND could decrease multiorgan injury-associated indicators, including serum creatine kinase, urea nitrogen, creatinine, lactate dehydrogenase, alanine aminotransferase, and aspartate aminotransferase, inhibit inflammatory cytokines IL-6 and TNF-α, regulate the serum levels of CRH, ACTH and CORT in LPS-induced septic rats, and alleviate the sepsis-induced morphological changes in the heart, liver, spleen, lung, and kidney and HPA tissues. SND had the ability to regulate the unbalanced glycerophospholipid metabolism, fatty acid β-oxidation, fatty acid amide metabolism, tryptophan metabolism and arachidonic acid metabolism to improve the LPS-induced sepsis. The results of western blotting analysis demonstrated that SND could decrease the expressions of LPCAT1 and IDO1 and increase the expressions of CPT1A and FAAH1 to regulate the above metabolic disorders. SND could alleviate LPS-induced sepsis partly via restoration of metabolic impairments in the HPA axis microenvironment, which provided important insights to future work to ascertain the mechanisms undergoing the HPA axis response to SND against sepsis.

Exploring the mechanism of Polygonum Cuspidatum in the treatment of ischemic stroke by network pharmacology analysis and experimental validation.

Polygonum cuspidatum (PC), a traditional Chinese medicine, has been employed in the treatment of ischemic stroke (IS), yet its precise mechanisms of action remain to be elucidated. This study integrates network pharmacology with in vitro and in vivo experiments to investigate the underlying mechanisms of PC in the treatment of IS. The active components of PC in rat serum samples were identified using ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). The active ingredients and mechanism of PC were predicted by network pharmacology. Interactions between key active compounds and major targets were validated through molecular docking. Finally, the efficacy and mechanism of PC in the treatment of IS were verified through in vivo and in vitro experiments. A total of 43 active components of PC were identified, along with 142 targets associated with IS. KEGG analysis suggested that the PI3K/Akt signaling pathway may play a key role in PC's regulation of IS-related injury. Molecular docking confirmed that quercetin, resveratrol, apigenin, luteolin, and torachrysone, may potentially interact with core targets including AKT1, IL6, TNF, TP53 and EGFR. Additionally, we confirmed that PC significantly inhibited the release of inflammatory cytokines and upregulated the phosphorylation of the PI3K/Akt signaling pathway in vitro. In vivo experiments further validated its protective effects. We speculate that this protective effect is mediated by the modulation of the PI3K/Akt signaling pathway.

Automatic identification of oligonucleotide metabolites in complex biological samples using ultra high-performance liquid chromatography high-resolution mass spectrometry combined with the molecule profiler software

Automatic identification of oligonucleotide metabolites in complex biological samples using ultra high-performance liquid chromatography high-resolution mass spectrometry combined with the molecule profiler software

Aqueous extract of Cornus officinalis alleviate NAFLD via protecting hepatocytes proliferation through regulation of the tricarboxylic acid cycle.

Cornus officinalis (CO) has been widely used as Chinese herbal medicine and has a good clinical efficacy in liver disease. In particular, it has a significant therapeutic effect on metabolic liver disease. However, systematic pharmacological studies on its hepatoprotective effect on non-alcoholic fatty liver disease (NAFLD) are lacking. We investigated the impact of Cornus officinalis extract (COE) on two mouse models of NAFLD, screened the potential mechanisms of action by using metabolomics assays, and explored the protective effects on hepatocyte proliferation by regulating glutamate metabolism and tricarboxylic acid (TCA) cycle. The main components of COE were identified by high performance liquid chromatograph (HPLC). Male C57BL/6J mice were subjected to construct carbon tetrachloride (CCl4) or methionine choline deficient (MCD) induced NAFLD mice. Liver function and lipid biochemical indicators were detected using commercial assay kits. Masson staining, Western blot, and immunohistochemistry analyses were used for assessing hepatic injury and fibrosis. LC-MS non-targeted analysis was performed using the 1290 Ultra-High Performance Liquid Chromatograph System and the 6540 Q-TOF Mass Spectrometry. Palmitic acid (PA) induced L-02cell model was established. The mediators in glutamate metabolism and TCA cycle were assessed by assay kits. In vivo experiments validated that COE significantly improved liver function in NAFLD mice, reduced lipid accumulation, and alleviated pathological damage and liver fibrosis. The non-targeted metabolomics analysis combined with Ingenuity Pathway Analysis (IPA) located glutamate metabolism and the downstream TCA cycle as potential mechanisms of COE, which was further confirmed in NAFLD model mice and PA-induced L-02cells. Finally, we confirmed that COE could promote mitochondrial energy supply by remodeling the homeostasis of the TCA cycle, thereby enhancing hepatocyte proliferation. This study demonstrated that COE could significantly improve CCl4 or MCD-induced NAFLD by promoting hepatocyte proliferation. These results highlighted the potential of COE as leads for the development of innovative treatments for NAFLD.

Integrated UHPLC-Q-exactive orbitrap HRMS and serum pharmacochemistry for the investigation of anti-hepatic fibrosis effect of Baoganning Decoction.

Early intervention in hepatic fibrosis (HF) is critical to reducing the risk of cirrhosis-related mortality and hepatocellular cancer. However, treating fibrosis has proven to be more challenging, with no approved anti-fibrotic therapies currently available for HF. Traditional Chinese medicines (TCMs) hold significant potential for the management of HF. This study aims to propose a systematic approach for investigating the pharmacological basis of Baoganning (BGN) Decoction, providing empirical evidence to support future research on its targets and mechanisms of BGN. Ultrahigh-performance liquid chromatography coupled with high- resolution mass spectrometry (UPLC-HRMS) was employed to analyze the chemical composition of BGN. Key compounds were investigated using disease databases to predict relevant targets, followed by molecular docking and molecular dynamics simulations to explore molecular-level interactions. The efficacy and critical targets of BGN were validated through in vivo and in vitro experiments. UPLC-HRMS was used to identify the chemical composition of the BGN, and serum pharmacology determined the active chemical constituents in rat plasma. Zebrafish, HSC-T6 cells, JS-1 cell line and mice served as experimental models to evaluate the antifibrotic effects of BGN. BGN demonstrated significant antifibrotic effect in vivo and in vitro models. A total of 757 compounds were identified in BGN, with 18 prototypical components and metabolites detected. Three compounds-quillaic acid, methyl cholate, and 3β-hydroxy-5-cholenoic exhibited dose-dependent inhibitory effects on HF. Molecular docking studies revealed stable interactions between these compounds and predicted targets. Additionally, the screened components effectively reduced the expression of ‌α-SMA and COL-I in both a cellular model and a zebrafish fibrosis model in a dose-dependent manner. The comprehensive analysis of BGN's chemical composition and its metabolic processes provides valuable insights into its pharmacological effects. These findings support the potential clinical and international application of BGN in treating hepatic fibrosis and improving patient outcomes.

Monitoring of occupational exposure to hazardous medicinal products in robotic compounding

ObjectiveThis study aims to evaluate the risk of occupational exposure to hazardous medicinal products (HMPs) when utilising robotic compounding systems for the preparation of antineoplastic sterile medications. Specifically, it assesses...

Organic Micropollutants in Waterways of a Large-Scale Water Diversion Project: Insights from Nontarget Screening and "Community" Analysis.

Large-scale water diversion projects are essential for meeting the needs of water-stressed regions, necessitating an evaluation of their impact on water quality and aquatic ecosystems. This study provides the first snapshots of organic micropollutants (OMPs) along the 1466 km Eastern Route of China's South-to-North Water Diversion Project. Using nontarget analysis with ultrahigh-performance liquid chromatography and high-resolution mass spectrometry, we identified and quantified 357 OMPs from water samples collected during the water diversion period (WDP) and the nonwater diversion period (NWDP). The OMPs included 136 household compounds, 112 agricultural compounds, 102 industrial compounds, and 7 traffic markers. Significant regional variations in OMP concentrations and compositions were observed during the NWDP due to diverse local pollution sources along the route. However, such differences were reduced during the WDP, likely due to water transfer. OMP diversity varied substantially during the NWDP but was more stable with less distance decay during the WDP. Network analysis indicated closer relationships between the OMPs during the WDP, suggesting a more consistent spatial distribution. The source water overwhelmingly influenced the water quality along the diversion route. These findings underscore the need for ongoing assessments of the impact of water diversion on regional water quality and ecosystems.

Identification of novel serum lipid metabolism potential markers and metabolic pathways for oral cancer: a population-based study

ObjectiveThis study aims to identify potential lipid biomarkers and metabolic pathways associated with oral cancer (OC). Then to establish and evaluate disease classification models capable of distinguishing OC patients from healthy controls.MethodsA total of 41 OC patients and 41 controls were recruited from a hospital in Southeast China to examine the serum lipidomics by Ultra-high Performance Liquid Chromatography Q Exactive Mass Spectrometry (UHPLC-QE-MS).ResultsThe total serum lipid profile showed that triglycerides accounted for the highest proportion of total metabolites, reaching 35.90% of the total. A total of 74 different metabolites were screened (12 up-regulated and 62 down-regulated), mainly enriched in the glycerophospholipid metabolism pathway. The three most significant changes in lipid metabolites were phosphatidylcholine (PC(18:3e/17:2)), acylcarnitine (ACar(14:2)), and glucuronosyldiacylglycerol (GlcADG(14:1/14:1)). The disease classification model, constructed using a KNN algorithm with 13 metabolites selected through LASSO screening, achieved the best performance, with an AUC of 0.978 (0.955-1.000).ConclusionLipid metabolic biomarkers identified in this study exhibit potential as candidate biomarkers for OC diagnosis. Further validation through prospective studies is required to confirm their clinical utility in early detection.

Optimized Enzymatic Extraction of Phenolic Compounds from Verbascum nigrum L.: A Sustainable Approach for Enhanced Extraction of Bioactive Compounds

Verbascum nigrum, commonly known as black mullein, is widely used in traditional medicine for its expectorant, mucolytic, sedative, and diuretic properties. This study aimed to develop and optimize a standardized method for extracting phenolic compounds from V. nigrum using enzymatic pretreatment followed by solvent extraction. Enzymatic treatment does not rely on harmful solvents and is a low energy-intensive process, making it a suitable green technology for the food, cosmetic, and pharmaceutical industries. The research explored the use of different lignocellulolytic enzymes, including pectinase, cellulase, α-amylase, and xylanase, to break down plant cell walls, enhancing the release and bioaccessibility of active compounds. The two-step extraction process proposed combined enzymatic pretreatment and hydroalcoholic extraction, resulting in a considerably improved yield of phenolic compounds (24 mg/g DM). Analytical characterization using a high-performance liquid chromatography (HPLC) system coupled with a diode-array-detector (DAD) and ultra-high-performance liquid chromatography (UHPLC) coupled with DAD and tandem mass spectrometry (MS/MS) revealed a higher concentration of target bioactive compounds in enzymatically treated extracts compared to traditional methods, including phenolic derivatives (e.g., caffeic acid, p-coumaric acid, and verbascoside), and flavonoids (e.g., luteolin). Up to 22 phenolic and flavonoid compounds were characterized. This study provides new insight into the potential of enzymatic extraction as a green and efficient alternative to conventional extraction methods, for the production of high-quality herbal products richer in (poly)phenolic compounds, highlighting its potential for industrial applications.

P-1264. Explore The Pharmacokinetic/pharmacodynamics Target Attainment (PTA) of cefoperazone/sulbactam

Abstract Background Cefoperazone/sulbactam (CPZ/SUL) are broad-spectrum antibiotics. Current dosage guidelines consider the renal elimination characteristics of SUL. However, with the risk of antimicrobial resistance, whether current dosage recommendation of CPZ/SUL achieves the desired probability of target attainment (PTA) is unclear. The purpose of this study is to compare the PTA between adjusted-dose (AD) CPZ/SUL and standard-dose (SD) CPZ/SUL therapy. Methods This observational study was conducted in a tertiary hospital in Taiwan during November 2023 to March 2024. Patients using CPZ/SUL (1:1 ratio) were prospectively enrolled. Two blood samples were collected during the dosing interval to measure CPZ and SUL concentrations by using ultra-high performance liquid-chromatography. Concentrations at the middle of the dosing interval (Cm) and before dosing (Ct) were calculated based on measured concentration. PTA was represented by the fraction of time the free concentration was above the minimum inhibitory concentration (fT >MIC). 50% fT >MIC was defined as Cm >MIC, and 100% fT >MIC was defined as Ct >MIC. AD group was defined as reduced CPZ/SUL doses according to the labeling, while SD group had no dose adjustment despite renal impairments. Results A total of 37 patients were enrolled (59.5% male, mean age, 75.4±16.0 years). The most common indication of CPZ/SUL was pneumonia (83.8%). There were 26 patients (70.3%) in the AD group and 11 patients (29.7%) in the SD group. The proportion of patients with 100% fT >MIC was lower in the AD group, as compared to the SD group (CPZ: 15.4% versus 54.5%, P< 0.01; SUL: 11.5% versus 72.7%, P< 0.01). Additionally, 7.7% of patients in the AD group compared to 45.5% of patients in the SD group achieved 100% fT >MIC for both CPZ and SUL (P< 0.01). Thirty patients (81.1%) received weekly vitamin K to prevent CPZ-related coagulopathy, and none had an international normalized ratio above 2. Conclusion PTA was inadequate in patients receiving CPZ/SUL based on current dosage adjustment recommendations. Future investigation is needed to determine if inadequate PTA is linked to worse clinical outcomes, especially in resistant pathogens. Disclosures All Authors: No reported disclosures

Ferula communis leaf extract: antioxidant capacity, UHPLC–MS/MS analysis, and in vivo and in silico toxicity investigations

IntroductionFerula communis has demonstrated an abundance of pharmacological and antioxidative qualities.MethodsThis study investigates the antioxidant activity of F. communis leaf aqueous extract, total polyphenol and flavonoid concentrations, and ultra-high-performance liquid chromatography (UHPLC) composition and then evaluates the toxicity of the plant’s leaves in vitro and in silico. The major compound of the studied extract, namely, p-hydroxybenzoic acid, was chosen for a molecular docking technique to discover the inhibition mechanism toward antioxidant proteins. In addition, a detailed molecular dynamics simulation was carried out to examine the thermodynamic stability of the produced intermolecular interactions. The antioxidant capacity of the extracts of F. communis was evaluated using 2,2-diphenylpicryl hydroxyl (DPPH) radical and ferric reducing antioxidant power (FRAP) procedures. Acute toxicity was tested on albino mice at doses of 200, 300, and 400 mg/kg.ResultsThe results show that the polyphenol and flavonoid contents of the extract are significant (0.257 ± 0.003 mg Eq AG/mg and 0.32 ± 0.04 mg Eq Q/mg, respectively). The antioxidant activity illustrates that the extracts have notable activity in DPPH and FRAP assays. The toxicity study revealed that the mice’s behavior, body weight, and organ weights (liver and kidneys) were unaffected by Ferula communis leaf extract administration compared to controls. UHPLC–tandem mass spectrometry (MS/MS) analysis of the extract highlights the presence of 11 compounds, the most abundant of which is p-hydroxybenzoic acid, representing 53.65%. The predicted pharmacokinetic characteristics of absorption, distribution, metabolism, excretion, and toxicity (ADMET) attest to the well-absorbed nature of the isolated compounds, with human intestinal absorption (HIA) varying from 42% for arbutin (M3) to 100% for ursolic acid (M4).ConclusionIn conclusion, the leaves of Ferula communis are a good source of natural antioxidants and phenolic compounds. Thus, this study demonstrates that this plant has a wide range of applications, including natural food preservatives, pharmaceuticals, and cosmetics, as evidenced by ongoing research.

Yunvjian decoction attenuates lipopolysaccharide-induced acute lung injury by inhibiting NF-κB/NLRP3 pathway and pyroptosis

IntroductionYunvjian (YNJ) decoction, a classic traditional Chinese medicine prescription for inflammatory diseases, has demonstrated good therapeutic effects in the clinical treatment of pneumonia. The aim of this study was to clarify the effective ingredients and mechanism of action of YNJ on lipopolysaccharide (LPS)-induced acute lung injury (ALI).MethodsThe effects of YNJ were evaluated in a mouse model of LPS-induced ALI and in LPS-treated MLE-12 murine lung epithelial cells and RAW264.7 macrophages in vitro. The mechanism of action of YNJ on these model systems was studied using RNA sequencing, immunohistochemical analysis, immunoblotting, immunofluorescence, ELISA, and polymerase chain reaction assays. Ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was applied to identify the absorbed components of YNJ.ResultsYNJ attenuated pulmonary damage in LPS-treated mice, as evidenced by reduced protein content in bronchoalveolar lavage fluid, decreased lung wet/dry weight ratio, and improved respiratory function. Analysis of pneumonia-related lung injury samples from patients in the Gene Expression Omnibus dataset GSE40012 indicated that NOD-like receptor protein 3 (NLRP3)-mediated pyroptosis was a primary mechanism in ALI. YNJ reduced the phosphorylation of nuclear factor-kappa B (NF-κB) and decreased the expression levels of lung NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cleaved caspase-1, and interleukin-1β levels (IL-1β) in vivo. Administration of YNJ-containing mouse serum increased cell viability and decreased malondialdehyde and reactive oxidative species contents in LPS-stimulated MLE-12 cells. YNJ-containing serum also decreased the secretion of tumor necrosis factor-α, IL-6, and IL-1β in LPS-stimulated RAW264.7 macrophages, and promoted macrophage polarization toward an M2 phenotype. A total of 23 absorbed components were identified in YNJ-containing serum. Among those, network analysis and in vitro experiments indicated that diosgenin, timosaponin BII, and mangiferin are anti-inflammatory active substances.ConclusionYNJ attenuates LPS-induced ALI in mice by inhibiting pyroptosis of lung epithelial cells and macrophages via suppression of the NF-κB/NLRP3 pathway. Our findings provide novel insights into the therapeutic effects of YNJ on ALI.

Carbon nanotubes stabilized in chitosan sponge (CNT-CS) as a new alternative clean-up sorbent for pesticide multiresidue determination in eggplants using QuEChERS and UHPLC-MS/MS.

The continuous development and application of pesticides in agriculture require robust multiresidue detection methods to guarantee food safety. This study introduces a novel method for multiresidue determination of pesticides in eggplants using the QuEChERS procedure, incorporating a clean-up step using carbon nanotubes stabilized in chitosan sponge (CNT-CS) and ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) for analysis. Upon identifying the optimal extraction conditions, various sorbents were assessed for their efficacy in the dispersive solid-phase extraction (d-SPE). Among these, the biosorbent CNT-CS emerged as the most efficient and cost-effective material, showing the best recovery results from spiked blank samples. The validation demonstrated that the method was adequate for analyzing 111 pesticides in eggplant samples with practical method quantification limits from 5 to 25µgkg-1. When applied to commercial eggplant samples, 27 pesticides were detected, demonstrating the importance of the proposed method. It is noteworthy that the conducted work represents the first report on the use of the biosorbent CNT-CS in the clean-up step of the QuEChERS method. The results obtained in the validation and application steps demonstrate that the proposed method is an excellent alternative for monitoring programs.

A Comparative Study of the Biological Properties of Eugenia uniflora L. Fruits and Leaves Related to the Prevention of Cardiovascular Diseases

Cardiovascular diseases (CVDs) remain the leading cause of death globally, emphasizing the need for effective preventive strategies. Plant-based foods, rich in phytochemicals, offer a promising potential in CVD prevention. This study investigated the antioxidant, anti-inflammatory, and antihypertensive properties of two Eugenia uniflora L. varieties (orange and purple pitanga) and their leaves. Their antioxidant activity was assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation scavenging activity assays, while their antihypertensive activity was evaluated through angiotensin-converting enzyme (ACE) inhibition. Their anti-inflammatory potential was determined via protein denaturation inhibition. Both fruit varieties exhibited similar bioactivities, with the purple variety showing a slightly higher activity, except in the DPPH and ABTS assays. The leaves consistently demonstrated the lowest activities across all assays. Free polyphenols, dominated by gallic acid, were quantified using µ-QuEChERS followed by ultrahigh-performance liquid chromatography (UHPLC-PDA). The orange variety contained the highest concentration of gallic acid (13.1 mg/100 g DW). These findings highlight the potential of Eugenia uniflora L. extracts as natural antioxidant, anti-inflammatory, and antihypertensive agents, suggesting their value in food, pharmaceutical, and cosmetic applications for promoting human health and preventing CVDs.

Nutraceutical Prospects of Pumpkin Seeds: A Study on the Lipid Fraction Composition and Oxidative Stability Across Eleven Varieties

The oxidative stability of nutritive and bioactive lipids is essential for their functionality. This study evaluated the potential of lipid fractions from pumpkin seeds obtained from eleven high-performing cultivars of Cucurbita maxima Duchesne, C. pepo L., and C. moschata Duchesne cultivated in Poland, aiming to evaluate their stability for nutraceutical applications. This study investigated the intrinsic relationship between chemical composition and oxidative stability to identify cultivars with promising functional potential and commercial value. The fatty acid, sterol, and lipid antioxidant profiles were characterized using gas chromatography (GC), GC–mass spectrometry (GC-MS), and ultra-high-performance liquid chromatography (UPLC), respectively. Antiradical activity was assessed via the DPPH assay, and oxidative stability was evaluated using differential scanning calorimetry (DSC). The oils exhibited high levels of polyunsaturated fatty acids (PUFAs) (59.5–68.6%), with n-6/n-3 fatty acid ratios ranging from 66.5 to 211.6. The lipid extracts contained up to 97.1% Δ7-sterols, while key antioxidants included squalene (616.6–3092.0 mg/kg) and γ-tocopherol (54.1–423.6 mg/kg). Notably, the C. pepo cultivar ‘Moonshine’ was the least abundant in these bioactive compounds. The carotenoid content ranged from 5.7 to 19.4 mg/kg across the extracts. Among the studied cultivars, ‘Show Winner’ and ‘Pink Jumbo Banana’ (C. maxima) stood out as promising candidates for nutraceutical applications due to their elevated levels of tocopherols, carotenoids, and squalene. A moderate n-6/n-3 fatty acid ratio (100–170), coupled with balanced levels of γ-tocopherol and squalene, was found to significantly enhance the oxidative stability of pumpkin seed lipids. These lipid fractions also show potential as stabilizing additives for oils rich in α-linolenic acid but deficient in natural antioxidants.

Systematic identification of the chemical components of leaves and roots of Didymocarpus heucherifolius Hand.-Mazz. based on UHPLC Q-Exactive Orbitrap MS technology coupled with molecular networking strategy

Didymocarpus heucherifolius Hand.-Mazz. is plant of the genus Didymocarpus wall. (Gesneriaceae). Species in this genus have long been used in folk medicine to treat various illnesses, including wounds, kidney stones, inflammations, asthma, influenza, etc. However, there are few studies on chemical components. Crude plant extracts are a complex mixture of several biologically active specialized metabolites. Therefore, a rapid and accurate identification and quantification are critical in phytochemical analysis. An efficient approach based on molecular networking strategy coupled with ultra high-performance liquid chromatography coupled with Q-Exactive Orbitrap tandem mass spectrometry (UHPLC Q-Exactive Orbitrap MS) was used to analyze the chemical components systematically. The chemical was identified using retention time, accurate molecular weight, parent peaks, fragment peaks, fragmentation characteristics, comparative analysis with the literature, reference standards, and standard databases, including mzVault. A total of 108 compounds (32 phenylpropanoids, 14 phenolic acid, 6 fatty acids, 14 organic acid, 9 flavonoid, 15 amino acids, and 28 other components) were identified in the leaves and roots of D. heucherifolius in which 84 and 84 compounds were found in the leaves and roots, respectively. The results showed that 68 components were different in plant roots and leaves, and these compounds were mainly represented in organic acids. Moreover, 50 identical chemical compounds were identified in plant roots and leaves, and these compounds were mainly phenylpropanoid compounds. The UHPLC Q-Exactive Orbitrap MS method was used for the first time to accurately, quickly, and systematically identify the compounds of the roots and leaves of Didymocarpus heucherifolius Hand.-Mazz., thereby providing a theoretical basis for its quality control, in-depth research, and further development.